Background
Short morning exposure to high illuminance visible electromagnetic radiations termed as artificial daylight is beneficial for the mental health of people living in geographical areas with important seasonal changes in daylight illuminance. However, the commercial success of high illuminance light sources has raised the question of the safety of long hour exposure.
Methods
We have investigated the effect of the replacement of natural daylight by artificial daylight in Swiss mice raised under natural lighting conditions. Mice were monitored for neurotoxicity and general health changes. They were submitted to a battery of conventional tests for mood, motor and cognitive functions’ assessment on exposure day (ED) 14 and ED20. Following sacrifice on ED21 due to marked signs of neurotoxicity, the expression of markers of inflammation and apoptosis was assessed in the entorhinal cortex and neurons were estimated in the hippocampal formation.
Results
Signs of severe cognitive and motor impairments, mood disorders, and hepatotoxicity were observed in animals exposed to artificial daylight on ED20, unlike on ED14 and unlike groups exposed to natural daylight or conventional lighting. Activated microglia and astrocytes were observed in the entorhinal cortex, as well as dead and dying neurons. Neuronal counts revealed massive neuronal loss in the hippocampal formation.
Conclusions
These results suggest that long hour exposure to high illuminance visible electromagnetic radiations induced severe alterations in brain function and general health in mice partly mediated by damages to the neocortex-entorhinal cortex-hippocampus axis. These findings raise caution over long hour use of high illuminance artificial light.
Highly active antiretroviral therapyi (HAART), a combination of drugs (lamivudine, zidovudine and nevirapine) used for pre-exposure prophylaxis and management of Human Immunodeficiency Virus infection in sub-Saharan Africa. The objective of this research work was to investigate the potential Ameliorative effect of neuroviteon on HAART induced toxicity on the cerebellum. Thirty two Wistar rats were divided into 4 groups of 8 rats each. Group A served as the control, while group B were administered with 9.28 mg/kg of HAART, group C received 9.28 mg/kg of HAART and 0.07mg/kg of folic acid and group D received 0.07mg/kg folic acid. Drugs were administered twice daily for 30 days after which neurobehavioural test of open field maze was perform. The rats were then sacrificed and their cerebellum harvested, processed and stained using haematoxylin and eosin method and nuro-fillament (NF) immunochemistry method. The slides were viewed under light microscope. Results showed a significant reduction in the brain to body weight index between the HAART group and the control and folic acid group. There was significant reduction in locomotor activity following administration of HAART to the animals compared with control, there were also significant reduction in rearing frequency , walling frequency and freezing duration, with a significant increased in freezing duration in the HAART treatment group. The freezing frequency, central line crossing and grooming frequency were not significantly different. The cerebella were affected with mild to moderate shrinkage of pyramidal cells and distortion of the granular cells. There was increased expression of NF in the HAART group compared to controls. HAART affects the weight, histology of the cerebellum and neurobehaviour. Neurovite has the potential of ameliorating the histological distortion and may be beneficial to people taking HAART.
POTENTIAL OF CURCUMIN IN PYRAMID CELL NECROSIS OF MICE (MUS MUSCULUS) DUE TO ...IAEME Publication
Mercury pollution in the aquatic environment can cause intoxication of marine
organisms and the formation of free radicals in the human body if consumed. Curcumin
is a natural ingredient that contains extracellular antioxidants to overcome the
formation of free radicals in the body. The purpose of the study was to determine the
effect of administering curcumin to the number of pyramid cells that were necrotic in
mice (Mus musculus) exposed to methylmercury. This experiment used a completely
randomized design with 4 groups of mice, i.e. 0.5 ml distilled water, 0.056 mg/kg
methylmercury, 0.0056 mg/kg methylmercury + 150 mg/kg curcumin and 0.0056 mg/kg
methylmercury + 300 mg/kg curcumin. Examination of total pyramid cell necrosis was
observed with a 400x magnification light microscope. In this study, there was a
decrease in the number of pyramid cell necrosis in mice. The decrease in the number
of necrotic cells was directly proportional to the increase in dose given. The treatment
results as positive controls showed the highest value with the number of pyramid cells
experiencing necrosis of 23.28. The highest decreases in the number of each treatment
were 10.32 in 150 mg/kgBW curcumin and 5.80 in 300 mg/kgBW curcumin. Curcumin
can reduce the number of pyramid cells that experience necrosis due to exposure to
methylmercury.
Highly active antiretroviral therapyi (HAART), a combination of drugs (lamivudine, zidovudine and nevirapine) used for pre-exposure prophylaxis and management of Human Immunodeficiency Virus infection in sub-Saharan Africa. The objective of this research work was to investigate the potential Ameliorative effect of neuroviteon on HAART induced toxicity on the cerebellum. Thirty two Wistar rats were divided into 4 groups of 8 rats each. Group A served as the control, while group B were administered with 9.28 mg/kg of HAART, group C received 9.28 mg/kg of HAART and 0.07mg/kg of folic acid and group D received 0.07mg/kg folic acid. Drugs were administered twice daily for 30 days after which neurobehavioural test of open field maze was perform. The rats were then sacrificed and their cerebellum harvested, processed and stained using haematoxylin and eosin method and nuro-fillament (NF) immunochemistry method. The slides were viewed under light microscope. Results showed a significant reduction in the brain to body weight index between the HAART group and the control and folic acid group. There was significant reduction in locomotor activity following administration of HAART to the animals compared with control, there were also significant reduction in rearing frequency , walling frequency and freezing duration, with a significant increased in freezing duration in the HAART treatment group. The freezing frequency, central line crossing and grooming frequency were not significantly different. The cerebella were affected with mild to moderate shrinkage of pyramidal cells and distortion of the granular cells. There was increased expression of NF in the HAART group compared to controls. HAART affects the weight, histology of the cerebellum and neurobehaviour. Neurovite has the potential of ameliorating the histological distortion and may be beneficial to people taking HAART.
POTENTIAL OF CURCUMIN IN PYRAMID CELL NECROSIS OF MICE (MUS MUSCULUS) DUE TO ...IAEME Publication
Mercury pollution in the aquatic environment can cause intoxication of marine
organisms and the formation of free radicals in the human body if consumed. Curcumin
is a natural ingredient that contains extracellular antioxidants to overcome the
formation of free radicals in the body. The purpose of the study was to determine the
effect of administering curcumin to the number of pyramid cells that were necrotic in
mice (Mus musculus) exposed to methylmercury. This experiment used a completely
randomized design with 4 groups of mice, i.e. 0.5 ml distilled water, 0.056 mg/kg
methylmercury, 0.0056 mg/kg methylmercury + 150 mg/kg curcumin and 0.0056 mg/kg
methylmercury + 300 mg/kg curcumin. Examination of total pyramid cell necrosis was
observed with a 400x magnification light microscope. In this study, there was a
decrease in the number of pyramid cell necrosis in mice. The decrease in the number
of necrotic cells was directly proportional to the increase in dose given. The treatment
results as positive controls showed the highest value with the number of pyramid cells
experiencing necrosis of 23.28. The highest decreases in the number of each treatment
were 10.32 in 150 mg/kgBW curcumin and 5.80 in 300 mg/kgBW curcumin. Curcumin
can reduce the number of pyramid cells that experience necrosis due to exposure to
methylmercury.
These are the abstracts indexed with my name in the Proceedings of the National Symposium on Evolving Paradigm to Improve Productivity from Dynamic Management and Value Addition for Plant Genetic Resources.
Isolated central nervous system tuberculosis is uncommon in immunocompetent patients. It resembles a pyogenic brain abscess
clinically and radiologically and poses a problem in diagnosis and treatment. Here we described a case of recurrent frontal lobe
abscess, which was diagnosed as a tubercular abscess. There was no clinical or radiological evidence of active tuberculosis
elsewhere in the body. The diagnosis of tubercular abscess was confirmed by Mycobacterium tuberculosis by Polymerase Chain
Reaction (TB-PCR) in the abscess material aspirated via a burr hole.
Study of anticonvulsant activity of quinidine in albino rats using pentylenet...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Hematological and Liver Function of Plasmodium Berghei Positive Wister Treate...ijtsrd
Eradication of malaria in Africa continues to be one of the greatest challenges in the health sector. All the drugs developed thus far have their limitations and are generally expensive. In Africa and Nigeria the use of herbs in treating sicknesses dated as far back as the existence of man and is still in use today by many Nigerians in rural areas. Here we evaluated the therapeutic efficacy of ethanolic extract for three of these herbs lemon grass, lime and turmeric used singly and in combination on Plasmodium berghei infected rats. Results reveals that there was an 81.54 reduction in parasite level in groups treated with herbs and an 88.9 reduction in those treated with ACTs. The time frame of this study wasn’t enough to determine if there will be a resurgence in the parasitemia level. Results from the liver function test also reveals that the herbs also reduced the levels of liver enzymes in the serum but the results from the liver histology from the onset shows little or no damage to the liver This helps us to understand that the plasmodium parasites does not cause much damage to the liver cells or requires more time to do so. Hence the study concludes that potent herbs like turmeric, lemon grass and lime although not as effective as ACTs but if harnessed properly can be substituted for ACTs in treating malaria in low income rural areas of Nigeria. Nwaisaac Ikechukwu Samuel | Igbokwe Vincent Ugochukwu | Okonkwo Chukwudi Onyeka | Okpa Precious Nwaka | Nnyaha Anthonia Ekenedilichukwu | Ajeka Prisca "Hematological and Liver Function of Plasmodium Berghei Positive Wister Treated With Herbs and Acts" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46277.pdf Paper URL: https://www.ijtsrd.com/medicine/other/46277/hematological-and-liver-function-of-plasmodium-berghei-positive-wister-treated-with-herbs-and-acts/nwaisaac-ikechukwu-samuel
Combination Therapy of Carica Papaya and Vernonia Amygdalina Leaf Extracts ar...ijtsrd
The study focused on the comparative determination of the effects of the combination of leaf extracts of Carica papaya CP and Vernonia amygdalina VA and Artemether based combination therapy ACTs on Plasmodium berghei infected male wistar rats. Fifty 50 male wistar rats, weighing 100 to 150g, were allotted into 5 groups n=7 . Group 2 negative control was infected but not treated, groups 3 5 which were infected received 250mg kg b.w each of CP and VA in combination and 4mg kg b.w of the two reference ACTs. Treatment was done orally once daily for 3 consecutive days, after which the animals were sacrificed and haematological analysis carried out. Afterwards, an observatory period of 7 days commenced after which another round of sacrifice was done.Before treatment, parasitemia count of animals in groups 3 5 was substantially p 0.05 higher when juxtaposed with group 2. AST and ALT activities were significantly p 0.05 elevated in group 3 5 when matched with group 2. Plasmodium berghei induction meaningfully p 0.05 lowered white blood cell WBC at all groups. After treatment, the extracts and drug which notably p 0.05 lowered plasmodium count, RBC, WBC, PCV, Hb and Platelet levels, did not meaningfully p 0.05 affect the activities of ALP, AST and ALT. After 7 days of observation, the extracts and drug efficaciously p 0.05 reduced plasmodium count, WBC and ALP activity further at all groups. These results indicate that the leaf extracts in combination are as efficacious as the ACTs. Ajeka Prisca Ojochogu | Igbokwe Ugochukwu Vincent | Okonkwo Onyeka Chukwudi | Nnyaha Anthonia. E. | Ikechukwu Samuel Nwaisaac | Okpa Precious Nwaka "Combination Therapy of Carica Papaya and Vernonia Amygdalina Leaf Extracts are as Efficacious as Acts" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46293.pdf Paper URL: https://www.ijtsrd.com/medicine/physiology/46293/combination-therapy-of-carica-papaya-and-vernonia-amygdalina-leaf-extracts-are-as-efficacious-as-acts/ajeka-prisca-ojochogu
Abstract
Objective(s):
Gold nanoparticles (GNPs) command a great deal of attention for biomedical applications nowadays. The data about the degree of toxicity and the accumulation of gold nanoparticles in-vivo is not enough to judge.
Materials and Methods:
A total of 32 healthy male Wistar rats were randomly divided into 4 including: three GNP-treated and one control group. Groups 1, 2 and 3 received 0.5 cc of a solution containing 5, 10, and 100 ppm Au daily via intraperitoneal (IP) injection for 7 days, respectively. The control group was treated with 0.5 cc normal saline with same procedure. Then, several biochemical parameters such as serum glutamate oxaloacetat transaminase (SGOT) and serum glutamate pyrvate transaminase (SGPT) were evaluated at 2, 7 and 14 days after the last injection. After 14 days, all the rats were sacrificed and liver, lung tissues were separated and evaluated.
Results:
SGOT two days after intervention was significantly greater in the group 2 than the control group. In liver histological assessment, in group 1, basophils were observed around the central veins, in group 2 fading and no observation of central veins was seen, and in group 3 hepatic damage was noticed. The lung histological results showed severe vascular hyperemia in group 1, air sacs damage in group 2, and complete air sacs destruction in group 3.
Conclusion:
The results showed extreme changes in the histopathology of lung and liver tissues caused by spherical nanogold with 5-10 nm size in all of three treatment groups.
Spatial Memory Disturbance Following Transient Brain Ischemia is Associated w...UniversitasGadjahMada
A number of studies have investigated the effects of ischemic injury on functional and cellular characteristics of hippocampus. There is only a limited study on vascular remodeling of it. The present study aimed at examining vascular remodeling in hippocampus and spatial memory disturbances after transient brain ischemia. Male Wistar rats were randomly divided into four groups, i.e. sham operated (SHAM), transient brain ischemia with 1 day reperfusion (IR1), 3 day reperfusion (IR3), and 10 days reperfusion (IR10) groups. Transient brain ischemia was induced by bilateral common carotid artery occlusion (BCCAO). The spatial memory test was performed using the Morris water maze (MWM) in SHAM and IR10 groups. The rats were euthanized at day 1, 3 or 10 after BCCAO depending on the groups. The mRNA expressions of SOD2, Bcl-2, NeuN, eNOS, endothelin-1 (ET-1), CD31, VE-cadherin and vascular remodeling of the hippocampus were examined. There were deteriorations of spatial learning ability in IR10 group. The percentages of SOD2 and Bcl-2, the expression of NeuN, decreased and the vascular remodeling was observed in the ischemic groups. The eNOS and CD31 expressions were less in IR10, the VE-cadherin expression was less in all ischemic groups than in SHAM group, while ET-1 expression in IR1 group was higher than any other groups. The spatial memory deterioration after BCCAO is associated with vascular remodeling in hippocampus, characterized by lumen narrowing and smooth muscle thickening of microvessels.
These are the abstracts indexed with my name in the Proceedings of the National Symposium on Evolving Paradigm to Improve Productivity from Dynamic Management and Value Addition for Plant Genetic Resources.
Isolated central nervous system tuberculosis is uncommon in immunocompetent patients. It resembles a pyogenic brain abscess
clinically and radiologically and poses a problem in diagnosis and treatment. Here we described a case of recurrent frontal lobe
abscess, which was diagnosed as a tubercular abscess. There was no clinical or radiological evidence of active tuberculosis
elsewhere in the body. The diagnosis of tubercular abscess was confirmed by Mycobacterium tuberculosis by Polymerase Chain
Reaction (TB-PCR) in the abscess material aspirated via a burr hole.
Study of anticonvulsant activity of quinidine in albino rats using pentylenet...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Hematological and Liver Function of Plasmodium Berghei Positive Wister Treate...ijtsrd
Eradication of malaria in Africa continues to be one of the greatest challenges in the health sector. All the drugs developed thus far have their limitations and are generally expensive. In Africa and Nigeria the use of herbs in treating sicknesses dated as far back as the existence of man and is still in use today by many Nigerians in rural areas. Here we evaluated the therapeutic efficacy of ethanolic extract for three of these herbs lemon grass, lime and turmeric used singly and in combination on Plasmodium berghei infected rats. Results reveals that there was an 81.54 reduction in parasite level in groups treated with herbs and an 88.9 reduction in those treated with ACTs. The time frame of this study wasn’t enough to determine if there will be a resurgence in the parasitemia level. Results from the liver function test also reveals that the herbs also reduced the levels of liver enzymes in the serum but the results from the liver histology from the onset shows little or no damage to the liver This helps us to understand that the plasmodium parasites does not cause much damage to the liver cells or requires more time to do so. Hence the study concludes that potent herbs like turmeric, lemon grass and lime although not as effective as ACTs but if harnessed properly can be substituted for ACTs in treating malaria in low income rural areas of Nigeria. Nwaisaac Ikechukwu Samuel | Igbokwe Vincent Ugochukwu | Okonkwo Chukwudi Onyeka | Okpa Precious Nwaka | Nnyaha Anthonia Ekenedilichukwu | Ajeka Prisca "Hematological and Liver Function of Plasmodium Berghei Positive Wister Treated With Herbs and Acts" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46277.pdf Paper URL: https://www.ijtsrd.com/medicine/other/46277/hematological-and-liver-function-of-plasmodium-berghei-positive-wister-treated-with-herbs-and-acts/nwaisaac-ikechukwu-samuel
Combination Therapy of Carica Papaya and Vernonia Amygdalina Leaf Extracts ar...ijtsrd
The study focused on the comparative determination of the effects of the combination of leaf extracts of Carica papaya CP and Vernonia amygdalina VA and Artemether based combination therapy ACTs on Plasmodium berghei infected male wistar rats. Fifty 50 male wistar rats, weighing 100 to 150g, were allotted into 5 groups n=7 . Group 2 negative control was infected but not treated, groups 3 5 which were infected received 250mg kg b.w each of CP and VA in combination and 4mg kg b.w of the two reference ACTs. Treatment was done orally once daily for 3 consecutive days, after which the animals were sacrificed and haematological analysis carried out. Afterwards, an observatory period of 7 days commenced after which another round of sacrifice was done.Before treatment, parasitemia count of animals in groups 3 5 was substantially p 0.05 higher when juxtaposed with group 2. AST and ALT activities were significantly p 0.05 elevated in group 3 5 when matched with group 2. Plasmodium berghei induction meaningfully p 0.05 lowered white blood cell WBC at all groups. After treatment, the extracts and drug which notably p 0.05 lowered plasmodium count, RBC, WBC, PCV, Hb and Platelet levels, did not meaningfully p 0.05 affect the activities of ALP, AST and ALT. After 7 days of observation, the extracts and drug efficaciously p 0.05 reduced plasmodium count, WBC and ALP activity further at all groups. These results indicate that the leaf extracts in combination are as efficacious as the ACTs. Ajeka Prisca Ojochogu | Igbokwe Ugochukwu Vincent | Okonkwo Onyeka Chukwudi | Nnyaha Anthonia. E. | Ikechukwu Samuel Nwaisaac | Okpa Precious Nwaka "Combination Therapy of Carica Papaya and Vernonia Amygdalina Leaf Extracts are as Efficacious as Acts" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46293.pdf Paper URL: https://www.ijtsrd.com/medicine/physiology/46293/combination-therapy-of-carica-papaya-and-vernonia-amygdalina-leaf-extracts-are-as-efficacious-as-acts/ajeka-prisca-ojochogu
Abstract
Objective(s):
Gold nanoparticles (GNPs) command a great deal of attention for biomedical applications nowadays. The data about the degree of toxicity and the accumulation of gold nanoparticles in-vivo is not enough to judge.
Materials and Methods:
A total of 32 healthy male Wistar rats were randomly divided into 4 including: three GNP-treated and one control group. Groups 1, 2 and 3 received 0.5 cc of a solution containing 5, 10, and 100 ppm Au daily via intraperitoneal (IP) injection for 7 days, respectively. The control group was treated with 0.5 cc normal saline with same procedure. Then, several biochemical parameters such as serum glutamate oxaloacetat transaminase (SGOT) and serum glutamate pyrvate transaminase (SGPT) were evaluated at 2, 7 and 14 days after the last injection. After 14 days, all the rats were sacrificed and liver, lung tissues were separated and evaluated.
Results:
SGOT two days after intervention was significantly greater in the group 2 than the control group. In liver histological assessment, in group 1, basophils were observed around the central veins, in group 2 fading and no observation of central veins was seen, and in group 3 hepatic damage was noticed. The lung histological results showed severe vascular hyperemia in group 1, air sacs damage in group 2, and complete air sacs destruction in group 3.
Conclusion:
The results showed extreme changes in the histopathology of lung and liver tissues caused by spherical nanogold with 5-10 nm size in all of three treatment groups.
Spatial Memory Disturbance Following Transient Brain Ischemia is Associated w...UniversitasGadjahMada
A number of studies have investigated the effects of ischemic injury on functional and cellular characteristics of hippocampus. There is only a limited study on vascular remodeling of it. The present study aimed at examining vascular remodeling in hippocampus and spatial memory disturbances after transient brain ischemia. Male Wistar rats were randomly divided into four groups, i.e. sham operated (SHAM), transient brain ischemia with 1 day reperfusion (IR1), 3 day reperfusion (IR3), and 10 days reperfusion (IR10) groups. Transient brain ischemia was induced by bilateral common carotid artery occlusion (BCCAO). The spatial memory test was performed using the Morris water maze (MWM) in SHAM and IR10 groups. The rats were euthanized at day 1, 3 or 10 after BCCAO depending on the groups. The mRNA expressions of SOD2, Bcl-2, NeuN, eNOS, endothelin-1 (ET-1), CD31, VE-cadherin and vascular remodeling of the hippocampus were examined. There were deteriorations of spatial learning ability in IR10 group. The percentages of SOD2 and Bcl-2, the expression of NeuN, decreased and the vascular remodeling was observed in the ischemic groups. The eNOS and CD31 expressions were less in IR10, the VE-cadherin expression was less in all ischemic groups than in SHAM group, while ET-1 expression in IR1 group was higher than any other groups. The spatial memory deterioration after BCCAO is associated with vascular remodeling in hippocampus, characterized by lumen narrowing and smooth muscle thickening of microvessels.
Case Studies in Home Cage Monitoring: Rodent Behavior, Circadian Biology and ...InsideScientific
Automated home cage behavioral monitoring is receiving increasing attention from the scientific community because of its benefits with regards to translational research, data replicability and animal welfare. In this webinar, Kenneth Dyar (Helmholtz Diabetes Center) and Joanna Moore (GSK) discuss how home cage monitoring can be used to reduce animal stress, optimize methodology and guide physiology and animal behavior research.
Dr. Kenneth Dyar
Passive locomotor activity monitoring for real-time circadian study design
Circadian clocks are fundamental determinants of physiology, behavior and health. For skeletal muscle, the circadian clock promotes insulin sensitivity and orchestrates rhythms of glucose, lipid, and amino acid metabolism. Physical activity synchronizes circadian clocks by altering body temperature and through distribution of various hormones and metabolites. Research suggests that misalignment of the ‘muscle clock’ plays an important pathophysiological role in metabolic disease. In this webinar, Dr. Kenneth Dyar highlights some examples of how the DVC system can be used for locomotor activity monitoring in order to evaluate circadian alignment before, during or after various dietary and pharmacological interventions.
Dr. Joanna Moore
Using home-cage monitoring to determine the impact of timed mating on male mouse welfare
The use of sterile male mice to induce pseudopregnancy in female mice assigned for the implantation of embryos is a vital component in the production of Genetically Altered Animals (GAA). This process involves swapping a genetically sterile male’s female companion for a new female. In this presentation, Dr. Joanna Moore discusses the use of home cage activity monitoring to evaluate the potential impact of this procedure on the welfare of male mice and how the impact of this intervention may be reduced. All animal studies were ethically reviewed and carried out in accordance with the Animals (Scientific Procedures) Act 1986 and the GSK Policy on the Care, Welfare and Treatment of Animals.
Key topics will include…
- Using home cage activity as a readout for animal welfare
- Using locomotor activity to optimize methodology and validate study design in real-time
- Pre-study screening of cohorts for outliers
OM: Advanced Bio-Well Practical Hands on Training - Feb.5-6-7/2016Krishna Madappa
OM: As our global and concentrated Bio-Well community grows, so should our dedicated training.
Dr.K.Korotkov & Krishna see this as an essential educational module for creating a next generation of Bio-Well professional users.
As the software platforms continue to amplify the non-linearity and our understanding of our multi dimensional presence, we are here to ennoble a seamless interface to this evolutionary level of Human Development.
Ultimately our singular purpose is to assist all our Bio-Well users globally to be the stewards of the advanced science of Bio-Electrography and inspire a generation towards the "Sciences of Optimal Human Excellence".
In recent years as a result of the observation that the toxic effects of boron and its
products have increased intensive studies have been initiated in our country and in the world
regarding its effects, especially in the central nervous system, digestive system and reproductive
system. The aim was to determine the histopathological changes caused by boric acid in rat testis,
liver, kidney and brain tissues by light microscopy after oral administration of toxic dose of acute
boric acid.
Histopathological effects of nanosilver (Ag-NPs) in liver after dermal exposu...Nanomedicine Journal (NMJ)
Objective(s):
With the advent of nanotechnology, significant progress has been made in the area of nanoscale materials such as nanosilver (Ag-Nps). These nanoparticles have a wide range of applications and been used for antimicrobial purposes for more than a century. However, little
attention has been paid to the toxicity of nanosilver wound dressing. This study was designed to investigate the possible histopathological toxicity of Ag-NPs in liver of mice during wound healing.
Materials and Methods:
A group of 50 female BALB/c mice of about 8 weeks were randomly divided into two groups: Ag-NPs and control groups (n=25). After creating similar wound on the backs of all animals, the wound bed was treated in Ag-NPs group, with a volume of 50 microliters of the nanosilver solution (10ppm) ,and in control group, with the same amount of distilled water. The experiment lasted for 14 days. Histopathaological samplings of liver were conducted on days 2, 7 and 14 of the experiment.
Results:
Histopathological studies demonstrated time-dependent changes in mice liver treated with Ag-NPs compared to control group. Some changes include dilation in central venous, hyperemia, cell swelling, increase of Kupffer and inflammatory cells.
Conclusion:
This study suggests that use of nanosilver for wound healing may cause a mild toxicity, as indicated by time-dependent toxic responses in liver tissue. However, this issue will have to be considered more extensively in further studies
Effect of naringenin on 3 np induced huntington’s disease like symptoms by es...IJARIIT
The main aim of this study was to investigate the effect of Naringenin, a flavonoid on 3-Nitropropionic acid (3-NP)-
induced Huntington’s disease like symptoms by estimations of motor co-ordination and behavioral parameters. 3-NP is an
irreversible inhibitor of complex II in the mitochondria. 3-NP-induced neurodegeneration has been widely used as an animal
model of Huntington’s disease (HD). It replicates the pathology of HD by causing oxidative stress. Naringenin is a polyphenolic
compound, a bioflavonoid, known to have a neuroprotective effect in a rat model of Alzheimer’s disease. In the present study,
the neuroprotective effect of Naringenin on 3-NP induced oxidative stress in the rat was determined by behavioral parameters.
Rats were induced with 3-NP (15 mg/kg) intraperitoneally for 21 days and rats induced with 3-NP were treated with Naringenin
(25mg/kg and 75mg/kg) for 21 days. 3-NP caused a decline in motor function in the neurological score, locomotor activity, and
impaired rotarod activity. Naringenin treatment significantly improved grip strength indicating an improvement in motor
performance, alterations in % spontaneous alternations. These findings suggest the antioxidant potential of Naringenin
flavonoid against 3-Nitropionic acid induced neurotoxicity. However, more investigations are required to elucidate the cellular
mechanisms of Naringenin against 3-Nitropropionic acid induced Huntington’s disease like symptoms.
Effects of electric field on histopathological study, electrical properties a...inventy
The present work was undertaken in order to investigate the effects of electric field (EF) of strength 50Hz-3KV/m on the histopathology, dielectric properties and liver function tests in albino rats. Fifty male albino rats were equally divided into three groups namely A, B, and C. Animals of group A used as control group which didn't receive any treatment . Animals of group B was divided into two subgroups namely B1 and B2 which were discretely exposed to 50HZ, 3KV/m electric field for a period of 15 day (8 hours/day, 5day/week). Group B2 animals were left to survive and housed at normal environmental conditions similar to control group A for a period of 15 day post exposed. Animals of group C are divided into two subgroups namely C1 and C2 were discretely exposed to the electric field for a period of 30 day (8 hours/day, 5day/week). Group C2 animals were left to survive and housed at normal environmental conditions similar to control group A for a period of 15 day post exposed. At the end of this period, blood and tissues samples were collected from all groups for experimental investigations. The dielectric constant (έ), electrical conductivity (σ) was measured in frequency range 42Hz-5MHz to investigate any changes in liver structure through studding histopathological examination. Also, the liver function was studied through analysis of glutamic oxaloacetic transaminase (GOT), glutamic pyruvie transaminase (GPT) and total protein (TP) after exposure to electric field this biochemical parameters have been evaluated in the blood serum of rats. The obtained results show high significant changes in the value of έ and σ of liver tissues for all groups exposed to EF as compared with control group. The levels of GOT and GPT were increased up to four times their values during the period of exposure to EF. These variations were recovered during two week after stopping exposure but they did not return to its original control values before exposure. On microscopic level; liver histological observations in liver cells which revealed some alterations including hepatic tissue with two portal tracts showing mild florous expansion and a dilated central vein, also ghosts of hepatocytes denoting necrotic changes also shows hepatic tissue with dilated central veins engorged with blood and splitting out to adjacent hepatocytes.
Ameliorative potential of the quercetin on lead-induced testicular damage mor...Prof. Hesham N. Mustafa
Background
Quercetin, a naturally occurring flavonoid known for its potent antioxidant properties, has been investigated for its potential in counteracting the harmful effects of lead (Pb) toxicity, which induces apoptosis and oxidative damage in various human tissues. This study aims to assess the reparative effects of quercetin on lead-induced testicular damage.
Methods
Four groups, each comprising ten adult male albino rats, were randomly assigned as follows: Quercetin group, Pb group, Pb + Quercetin group, and control group. All treatments were administered orally via gavage daily for a duration of 30 days. Evaluation of sex hormone levels (serum testosterone, FSH, and LH), cytokines and inflammatory mediators (IL-1β, TNF-α, MCP-1), lead concentration, oxidative and antioxidant stress markers (superoxide anion [O2−], MDA, SOD, CAT, GSH), and sperm characteristics were carried out.
Results
The results demonstrated a significant decline in sex hormones and antioxidants, accompanied by an increase in lead concentrations, cytokines, inflammatory mediators, and oxidative stress indicators (O2−, MDA), while SOD, CAT, and GSH levels were reduced. The Pb-intoxicated group exhibited a substantial increase in dead and abnormal sperm, along with significant reductions in sperm concentration and motility. Morphometrically, a marked decrease was observed in spermatogonia, primary spermatocytes, spermatids, and sertoli cells per seminiferous tubule, as well as epithelial height. Furthermore, coadministration of quercetin exhibited notable benefits. It significantly elevated testosterone levels (P < 0.001), testicular SOD, CAT, and GSH activities, while decreasing MDA levels (P < 0.001). Quercetin also mitigated the deleterious effects of lead toxicity on sperm parameters and restored morphometric variations, including epithelial height.
Conclusions
Quercetin supplementation alongside lead exposure showed a potential for ameliorating degenerative changes caused by lead toxicity in the testicles. This cotreatment effectively reduced oxidative stress, cytokine levels, inflammatory mediators, and restored biochemical alterations, thereby improving morphometric parameters.
The pattern of branching and intercommunications of the musculocutaneous nerv...Prof. Hesham N. Mustafa
Background:
The aim of the present work was to provide evidence about the anatomical variations as regard the origin, distribution, and branching pattern of the musculocutaneous nerve (MCN).
Materials and methods:
Brachial plexus was dissected in 40 upper limbs of 20 male adult cadavers. The pattern of the musculocutaneous nerve was photographed by a digital camera.
Results:
The location and length of the nerve branches between left and right arms were recorded and statistically analyzed. In (90%) of specimens the MCN originates from the lateral cord of the brachial plexus, in (5%) it arose from the median nerve (MN), while in the remaining (5%) specimen, it was absent. The musculocutaneous nerve pierced the coracobrachialis muscle in 90% of specimens, and in the remaining (10%) did not pierce it. The motor branches to biceps brachii muscle were categorized into: Type 1 (90%): one branch that divides to supply the two heads of biceps; Type 2 (5%): double branches, innervating each head of biceps separately. The motor branches to brachialis muscle were categorized into: Type 1 (82.9%): one branch; Type 2 (14.2%): double branches and Type 3 (2.9%): three branches that innervating brachialis muscle. Communications between the MCN and the MN were observed in 35% of specimens.
Conclusions:
The knowledge of the common and uncommon musculocutaneous nerve variations is important especially to the surgeons for carrying out surgical procedures in axilla and arm.
Morphohistometric analysis of the effects of Coriandrum sativum on cortical a...Prof. Hesham N. Mustafa
Objective: Natural compounds can act as metal chelators and oxygen free radical scavengers, which allows them to be used as bioactive antagonists to heavy metals neurotoxicity. The aim of the study to analyze the morphometric effects of Coriandrum sativum (C. sativum) on lead-induced neurotoxicity.
Materials and Methods: Forty Sprague-Dawley albino rats were divided into four equal groups (ten in each group): control group; coriander group: received aqueous C. sativum extracts (600 mg/kg BW for 60 days orally); lead (Pb) group: received a daily dose of lead acetate (Pb) (10 mg/kg BW for 60 days orally); Pb+ coriandrum group: received: aqueous C. sativum extract (600 mg/kg BW) prior to 10 mg/kg BW of Pb. The following parameters malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were measured. Layers thickness and nuclei density were analyzed.
Results: Lead levels in blood and tissues were decreased significantly in the Pb group and those findings were corrected significantly (p=0.001) with C. sativum addition. Data exhibited an increase in oxidative stress marker MDA and a decrease in antioxidant enzymes activities (SOD, CAT, and GPx) significantly in the Pb group and those effects were reversed significantly (p=0.001) by C. sativum administration. The cerebellar cortex and all layers of the somatosensory cortex thickness and nuclei density were diminished significantly in the Pb group. The morphometrical measurements were corrected significantly (p=0.001) by C. sativum.
Conclusion: From the findings of the current study, Pb caused noticeable structural and functional variations in the cerebellar cortex and somatosensory cortex. C. sativum corrected these parameters as it possesses chelating and antioxidant potentials.
Background:
The anterolateral ligament (ALL) is a true well-defined ligament in the knee first described in 1879 by Segond. After the work of Claes et al., several studies were conducted about biomechanics and its role in stability of the knee. The anatomical existence of the ALL has been studied by and various radiographic diagnostic modalities and in cadavers. It originates from lateral femoral epicondyle and is inserted between Gerdy’s tubercle and the fibular head. There has been controversy about the existence of ALL in pediatric patients. The aim of this work was to confirm the presence of ALL in pediatric patients by using MRI.
Materials and Methods:
We reviewed the knee MRI scans of 100 pediatric patients (ages between one and 12 yr) who had no knee injury or congenital deformity and had been evaluated by an expert radiologist.
Results:
The ALL was detected in 90% of the pediatric patients with the use of MRI.
Conclusions:
The main finding of this study was that ALL can be seen in pediatric patients using MRI. Despite numerous studies, additional research is needed to further define the role of the ALL in knee function.
Level of Evidence:
Level IV.
Protective effect of garlic extract against maternal and fetal cerebellar dam...Prof. Hesham N. Mustafa
Background: In spite of its industrial usefulness and varied daily uses, lead (Pb) pollution is a widespread ecological problem that faces the humans in the 21th century. Pb was found to produces a wide range of toxic effects including neurotoxicity especially to the developing and young offspring. Recently, the utilisation of herbal plants has received a significant attention where there has been rising awareness in their therapeutic use; among these is the garlic. In light of the above, the current study is designed experimentally in female pregnant rats in order to investigate the beneficial role of garlic extract in the protection from the maternal and foetal cerebellar damage produced by administration of different doses of Pb during pregnancy.
Materials and methods: Positively pregnant female rats were divided into five groups; one control group, two Pb-treated groups (exposed to 160 and 320 mg/kg b.w. of Pb, respectively) and two groups treated with both Pb and garlic (exposed to Pb as previous groups together with 250 mg/kg b.w./day of garlic extract). Treatments started from day 1 to day 20 of pregnancy, where the mother rats of different experimental groups were sacrificed to obtain the foetuses. Pb level in the maternal and foetal blood and cerebellum was estimated by spectrophotometry. Specimens of the cerebellum of different mother and foetal groups were processed to histological and immunohistochemical staining for microscopic examination.
Results: The results showed that administration of Pb to pregnant rats resulted in a dose-dependent toxicity for both mothers and foetuses in the form of decrease in maternal weight gain, placental and foetal weights, brain weight and diminished foetal growth parameters, which were prominent in rat's group treated with larger dose of Pb. In Pb-treated rats, Pb level in blood and cerebellum was high when compared with the control group. The histopathological examination of the cerebellum of treated dams and foetuses showed marked alterations mainly in the form of Purkinje cell degeneration and lack of development of foetal cerebellum. Co-treatment of garlic extract along with Pb resulted in a significant decrease in Pb levels as compared with those treated with Pb alone with improvement of the histopathological changes.
Conclusions: This study was useful in evaluating the hazardous effects of uncontrolled use of Pb in general and in assessing the developmental and neurotoxicity of foetuses due to exposure during pregnancy in particular. Co-administration of garlic has beneficial effects in amelioration of Pb-induced neurotoxicity and reversing the histopathological changes of the cerebellum of mother rats and foetuses. (Folia Morphol 2018; 77, 1: 1-15).
Keywords: Purkinje cells; garlic; glial fibrillary acidic protein; lead.
Protective role of co q10 or l carnitine on the integrity of the myocardium i...Prof. Hesham N. Mustafa
Doxorubicin (DOX) is a chemotherapeutic agent used for treatment of different cancers and its clinical usage is hindered by the oxidative injury-related cardiotoxicity. This work aims to declare if the harmful effects of DOX on heart can be alleviated with the use of Coenzyme Q10 (CoQ10) or L-carnitine. The study was performed on seventy two female Wistar albino rats divided into six groups, 12 animals each: Control group; DOX group (10mg/kg); CoQ10 group (200mg/kg); L-carnitine group (100mg/kg); DOX+CoQ10 group; DOX+L-carnitine group. CoQ10 and L-carnitine treatment orally started 5days before a single dose of 10mg/kg DOX that injected intraperitoneally (IP) then the treatment continued for 10days. At the end of the study, serum biochemical parameters of cardiac damage, oxidative stress indices, and histopathological changes were investigated. CoQ10 or L-carnitine showed a noticeable effects in improving cardiac functions evidenced reducing serum enzymes as serum interleukin-1 beta (IL-1 β), tumor necrosis factor alpha (TNF-α), leptin, lactate dehydrogenase (LDH), Cardiotrophin-1, Troponin-I and Troponin-T. Also, alleviate oxidative stress, decrease of cardiac Malondialdehyde (MDA), Nitric oxide (NO) and restoring cardiac reduced glutathione levels to normal levels. Both corrected the cardiac alterations histologically and ultrastructurally. With a visible improvements in α-SMA, vimentin and eNOS immunohistochemical markers. CoQ10 or L-carnitine supplementation improves the functional and structural integrity of the myocardium.
Keywords: Cardiotoxicity; CoQ10 and L-carnitine; Dox; Vimentin; eNOS.
One year mortality rate after hip fracture in the western region of saudi ara...Prof. Hesham N. Mustafa
Background:
The mortality rate of elderly patients who sustain a hip fracture is high compared to the general population. Identifying risk factors can help predict patients at risk of hip fracture to reduce the mortality rate. No studies have shown the mortality rate of patients with hip fractures in the western region of Saudi Arabia. Therefore, this study aimed to identify the risk factors associated with the mortality of patients with hip fractures admitted to the King Abdulaziz Hospital and compare the results with other studies.
Methods:
The mortality rate (within 1 yr or less) in 177 patients over the age of 60 yr who were admitted to the university hospital between July, 2007, and September, 2012, with hip fractures was retrospectively studied. The patients were assessed with regard to gender, age, type of hip fracture, and type of surgical intervention.
Results:
The overall mortality rate 1 yr after hip fracture was 12.43%, and the mean age was 77.77 yr old. The risk factors most associated with mortality were as follows: advanced age (71 to 80 and 81 to 90 yr old), male, peritrochanteric (extracapsular) fracture, and operative fixation with dynamic hip screw.
Conclusions:
The mortality rate of patients with hip fractures within 1 yr has a high-risk potential, especially for male patients over 71 yr of age with peritrochanteric (extracapsular) fractures. Surgical treatment with dynamic hip screw also was shown to be a risk factor between the different treatment options.
Level of Evidence:
Level IV.
Biomarkers of Systemic Lupus Erythematosus and Systemic Sclerosis diseases ac...Prof. Hesham N. Mustafa
Systemic Lupus Erythematosus (SLE) and systemic sclerosis (SSc) are systemic inflammatory autoimmune disorders characterized by a large spectrum of clinical and laboratory features. The aim of the present study was to investigate the possible use of serum level of soluble intercellular adhesion molecule-1(sICAM-1) and soluble interleukin-2 receptor (sIL-2Ra) as biomarkers for monitoring of SLE and SSc disease activity. Moreover, it aimed to compare the specificity and sensitivity as well as cut-off value of both biomarkers in a sample of Egyptian patients. 50 SLE patients, 30 SSc patients and 60 age and sex matched healthy controls were enrolled in our study. sICAM-1and sIL-2Ra were measured in serum samples obtained from all participants. In addition to Erythosedimentation rate (ESR), complete blood count (CBC), Antineuclearantibodies (ANA) estimation, disease activity of both diseases were also assessed. sICAM-1and sIL-2Ra levels were higher in SLE and SSc patients versus control. Both parameters are correlated with each other as well as the activity parameters. A cut-off levels of 455.59 (ng/ml) &2525935 (pg/ml) in both SLE & SSs respectively was observed with the highest specificity and sensitivity. It could be concluded that sICAM-1 and sIL-2Ra are noninvasive biomarkers for SLE and SSc that could play a pathophysiologic role in development and progression of both diseases. Moreover, sICAM-1 and sIL-2Ra are correlated with the disease activity at cut-off values of 455.59 (ng/ml) & 2525935(pg/ml) respectively.
Morphohistometric study of the ligamentum flavum in cervical,thoracic and lum...Prof. Hesham N. Mustafa
ABSTRACT Anatomic characterization and fine structure of the human ligamentum flavum (LF), especially at different spinal levels, represent an attractive focus for the scientific and surgical application. Descrip-tive anatomical and structural study of LF at the cervical, thoracic and lumbar levels of the vertebral column in human cadavers is carried out here. The aim of the work is to clarify the anatomical features and fine structural differences in the human LF at different vertebral levels (cervical, thoracic and lumbar). Specimens of vertebral column were ob-tained from 34 human preserved cadavers. Their average age ranged between 56 and 69 years. Morphometric parameters including height, width and thickness of the ligament flavum at the mid-levels of cervical, thoracic and lumbar regions were measured. Sections obtained from different levels were stained with different stains. Morpho-metric measurements involved the relative elastic area, relative collagen area, elastic area% and collagen area% were measured.The results of the height, width and thickness of the LF at different spinal levels showed gradual increase in their mean values respectively. The LF midline gaps were found in the cervical, thoracic and lumbar regions. The morphometrical measure-ments showed that the average elastic area was highest in the cervical region and lowest in the tho-racic region. In the lumbar region, the percentages of both elastic area and the collagen area were nearly the same. The characterization of morpho-logical and histological aspects of the LF at differ-ent spinal levels will be of great importance for ap-plications in spinal surgery, biomechanical and physical rehabilitation of vertebral column.Keywords: Ligamentum Flavum – Spinal – Collagen and elastic fibers
Correlation between acl injury and involvement of the anterolateral ligament ...Prof. Hesham N. Mustafa
Background:
Clinical testing has demonstrated the role of the anterolateral ligament (ALL) in controlling anterolateral laxity and knee instability at high angles of flexion. Few studies have discussed the association between an anterior cruciate ligament (ACL) injury and ALL injury, specifically after residual internal rotation and a post-ACL reconstruction positive pivot-shift that could be attributed to ALL injury. The goal of this study was to assess the correlation between ALL injury and ALL injury with concomitant ACL injury using MRI.
Material and Methods:
This was a retrospective study of 246 patients with unilateral ACL knee injuries from a database that was reexamined to identify whether ALL injuries occurred in association with ACL injuries. We excluded the postoperative reconstructed cases. The charts were reviewed on the basis of the presence or absence of diagnosed ACL injury with no regard for age or sex.
Results:
Of the 246 patients with ACL injury, there were 165 (67.1%) patients with complete tears, 55 (22.4%) with partial tears, and 26 (10.6%) with sprains. There were 176 (71.5%) patients with ALL and associated ACL injuries, whereas 70 (28.5%) did not have associated ACL injuries. There was a significant statistical relationship between ACL and ALL injuries (P<0.0001).
Conclusions:
There is high incidence of ALL tears associated with ACL injuries. Clinicians should be aware of this injury and consider the possibility of simultaneous ALL and ACL repair to prevent further knee instability.
Level of Evidence:
Level IV.
Liver ischemia/reperfusion injury, a setting in which the functional mass is ...Prof. Hesham N. Mustafa
Liver ischemia reperfusion is induced during sur-gical procedures like liver transplantation and re-section. Multiple mechanisms have been postulat-ed to liver damage following liver ischemia reperfu-sion injury, such as oxidative stress and inflamma-tory reactions. The present study declares the pos-sible mechanism of tadalafil, toward modulating the inflammatory response. Forty-eight rats were divided into 4 groups as follows; Sham group sub-jected to midline laparotomy only. Tadalafil group administered Tadalafil 10 mg/kg intraperitoneal 45 min before sham operation. I/R (Ischemia-reperfusion) group, rats undergo 60 min of hepatic ischemia followed by 60 min of reperfusion. Tada-lafil + I/R group rats undergo a similar pattern of I/R after the treatment with Tadalafil 10 mg/kg, 45 min before ischemia. At the end of the reperfusion, the blood samples were collected for estimation of biochemical markers including liver enzymes using colorimetric assay method and serum: TNF-α (tumor necrosis factor-α), IL-6 (interleukin 6) le-vels, ICAM- 1 (Intercellular Adhesion Molecule-1) were measured. Tissues were evaluated by semi-quantitative and morphometrical approaches. Ta-dalafil succeeded in restoring normal levels of liverenzymes and ameliorating the oxidative stress as evidenced by decreasing MDA and restoring redu-ced glutathione levels in liver tissue homogenate. Also, Tadalafil exhibits anti-inflammatory effects, as it significantly decreased the levels of TNF-α, IL6 and ICAM-1. The findings are supported by BCL-2, TNF-α immunomarkers. It is concluded that modulation of the inflammatory response might be one of the mechanisms of Tadalafil-mediated he-patoprotection, so it is recommended as an adju-vant therapy in liver surgery.Keywords: Ischemia/reperfusion injury – Oxidative stress – Apoptosis – TNF-α – BCL-2
Neuro-amelioration of cinnamaldehyde in aluminum-induced Alzheimer’s disease ...Prof. Hesham N. Mustafa
Aluminum (Al) is a neurotoxic substance which has played an important role in the etiology, pathogenesis, and development of amyloid-β (Aβ) plaques. This study was carried out to evaluate the neuroprotective effect of aqueous cinnamon extract against aluminum chloride (AlCl3)-induced Alzheimer’s disease. Forty adult male albino rats, randomly divided into four equal groups. Control group; ACE200 group administered aqueous cinnamon extract (ACE) orally; AlCl3 group received daily intraperitoneal (i.p.) injection of AlCl3 for 60 days to induce neurotoxicity and AlCl3 + ACE200 group received a combination of AlCl3 and ACE in the same dose and route as previous groups. Aluminum administration significantly enhanced the memory impairment and the Aβ formation in the rat model. The cerebellum exhibited a significant reduced number of Purkinje cells, marked decrease in the density of dendritic arborization and prominent perineuronal spaces in the molecular layer. There was loss of dendritic spines, neurofibrillary degeneration, and appearance of neuritic plaques. Concomitant administration of AlCl3 and ACE displayed an observable protection against these changes with progressive improvement in memory and intellectual performance. In conclusion, ACE may play a protective role against formation of amyloid-β plaques in cerebellum.
Analytical Study of Clinicopathological Data of Saudi Patients with Osteoarth...Prof. Hesham N. Mustafa
SUMMARY: Knee osteoarthritis (OA) is a common disabling disease. Epidemiological studies have revealed various risk
factors for OA, including sex, aging, obesity, occupational illnesses, and chronic diseases. Here we evaluate the clinical, pathological,
and radiological findings of knee OA in a subset of Saudi patients who were subjected to total knee replacement (TKA). The study
population included 30 Saudi patients with knee OA who were operated by TKA (from June 2014 to December 2015) in the Department
of Orthopedics, Faculty of Medicine, King Abdulaziz University, Saudi Arabia. Patient’s clinical and radiological data were collected
from the hospital files. Pathological examination of the excised superior articular surface of tibia and femoral condyles were done.
Pearson Chi-squared analysis was used to test for differences between the variables in associated risk factors. There were more women
than men. Sixty per cent of patients were older than 60 years [mean age, 59.2 (females) and 61.7 (men) years-old]. All patients exceeded
obesity class 1, with females being more obese than males. Pathological examination of the superior articular surface of tibia and femoral
condyles showed high score lesions, which was more apparent in females than in males. Radiological findings showed that most lesions
were high grade. The findings of this study will help to understand the pathogenesis of OA and improve treatment decision making
relevant to TKA in knee OA in Saudi Arabia and elsewhere.
KEY WORDS: Osteoarthritis; Knee; Arthroplasty.
A Study on the Toxic Effect of Different Doses of Diclofenac Sodium on the De...Prof. Hesham N. Mustafa
SUMMARY: The toxic effects of different doses of diclofenac sodium (DS) on the kidney on the postnatal period (0-7 days) by
morphometrical and immunohistochemical methods were investigated. For this purpose, 15 female adult wistar albino rats were used and
divided into 5 main groups. Group Ia served as normal control, physiologic group Ib received normal saline, group II received low dose (3.9
mg/kg), group III received medium dose (9 mg/kg) and group IV received high dose (18 mg/kg). Male offspring’s from 0-7 days after birth
were used in this study. On the 8th day of postnatal life, all animals were anesthetized. Then, the kidney samples were analyzed. Haematoxylin
and eosin staining showed degeneration and necrosis, apparent atrophy of the glomeruli, mononuclear cell infiltration, congested vessels,
increased fibrous tissue and distortion of the proximal convoluted tubules with interruption of the brush margin of the DS treated group.
Increased level of Caspase-3 and upregulation of TNF-α with different doses of DS. In light of our findings, DS may lead to adverse effects
that are dose-dependent in the prenatal subjected kidney to this drug.
KEY WORDS: Diclofenac sodium; Proximal convoluted tubules; Apoptosis;Cyclooxygenase.
Thymoquinone ameliorates oxidative damage and histopathological changes of de...Prof. Hesham N. Mustafa
ABSTRACT
Lead (Pb) toxicity is known to be a chief environmental health issue, especially for pregnant
women and young children. Today, the use of medicinal herbs in the treatment of many diseases
and different toxic agents has become highly accepted due to their effectiveness and lower costs.
Thymoquinone (TQ), which is extracted from Nigella sativa seeds, is a potent antioxidant and anti
inflammatory agent. This study was designed to explore the optional protectivity of TQ against
maternal and fetal oxidative stress and brain damage induced by Pb administration. Pregnant
rats were distributed into seven groups: control group, TQ group, DMSO group, two groups Pb
treated (160 and 320 ppm), and two groups Pb-treated (160 and 320 ppm) co-treated with TQ.
Administration started from gestation day 1 (GD1) to day 20 (GD20) through oral gavage once
daily. Lead administration caused a dose-dependent toxicity for both mothers and fetuses. Also,
the histopathological assessment of the brains from Pb-treated groups showed marked altera
tions. Co-treatment of with TQ and Pb caused a significant decrease in Pb levels as compared
with those treated with Pb alone and amelioration of histopathological changes in the brains. It
was concluded that co-treatment of TQ along with gestational Pb exposure could mitigate the
effects against Pb-induced maternal and fetal neurotoxicity.
KEYWORDS
Lead; oxidative stress; brain;
Thymoquinone; fetal toxicity
A study on the toxic effect of different doses of Diclofenac sodium on the de...Prof. Hesham N. Mustafa
The toxic effects of different doses of diclofenac sodium (DS) on the kidney on the postnatal period (0-7 days) by morphometrical and immunohistochemical methods were investigated. For this purpose, 15 female adult wistar albino rats were used and divided into 5 main groups. Group Ia served as normal control, physiologic group Ib received normal saline, group II received low dose (3.9 mg/kg), group III received medium dose (9 mg/kg) and group IV received high dose (18 mg/kg). Male offspring’s from 0-7 days after birth were used in this study. On the 8th day of postnatal life, all animals were anesthetized. Then, the kidney samples were analyzed. Haematoxylin and eosin staining showed degeneration and necrosis, apparent atrophy of the glomeruli, mononuclear cell infiltration, congested vessels, increased fibrous tissue and distortion of the proximal convoluted tubules with interruption of the brush margin of the DS treated group. Increased level of Caspase-3 and upregulation of TNF-α with different doses of DS. In light of our findings, DS may lead to adverse effects that are dose-dependent in the prenatal subjected kidney to this drug.
Keywords : Diclofenac sodium; Proximal convoluted tubules; Apoptosis; Cyclooxygenase.
Zingiber Officinale Alleviates Maternal and Fetal Hepatorenal Toxicity Induce...Prof. Hesham N. Mustafa
This study was designed to address the protective effects of Zingiber officinale on the toxic outcomes of prenatal Cadmium administration on pregnancy outcome. Pregnant female Sprague-Dawley rats were randomly divided into four groups (eight rats/each), control group received distilled water, 2nd group treated with 8.8 mg of CdCl2/kg b. wt, 3rd group treated with 250 mg of Zingiber officinale/kg b. wt, and 4th group treated with 250 mg of Zingiber officinale/kg b. wt, followed by 8.8 mg of CdCl2/kg b.wt. Daily body weight of pregnant was recorded from GD1-GD20, and then pregnant rats were sacrificed at GD20. Samples of maternal and fetal livers and kidneys were processed for histological examination. Administration of Cd to pregnant rats showed adverse effects on pregnant mothers and their fetuses; reduced maternal weight gain, reduced absolute organ weights, reduced fetal growth parameters and placental weights together with altered histological appearance of the maternal and fetal livers and kidneys. While co-administration of Zingiber officinale showed an improvement of these toxic alterations. Zingiber officinale through its antioxidant activity could be beneficial against toxic outcomes of Cd exposure during pregnancy.
The Ameliorative Potential of Dexmedetomidine and Benincasa Cerifera Extract ...Prof. Hesham N. Mustafa
Renal ischemia/reperfusion injury (IRI) represents the main reason for acute kidney injury (AKI). Dexmedetomidine (Dex) and Benincasa cerifera (BC) have wide benefits due to their anti-inflammatory and antioxidant properties. This study aims to illustrate the protective effects of BC and Dex on renal IRI in a diabetic model. Sixty adult male albino rats (Wistar strain), weighing 250–300 g, were included in the study. The rats were divided into four groups, as follows: sham group: (non-diabetic); diabetes mellitus (DM) + IRI group: streptozotocin (STZ)-induced diabetic rats exposed to renal IRI on day 30 after diagnosis of diabetes; DM + IRI + BC group: STZ-induced diabetic rats treated with BC (500 mg/kg) for 30 days after diagnosis of diabetes, then exposed to renal IRI; and DM + IRI + Dex group: STZ-induced diabetic rats treated with Dex (100 µg/kg intraperitoneally) 5 min before induction of ischemia on day 30 after diagnosis of diabetes, then exposed to renal IRI. Biochemical parameters, histopathological examination, and immunohistochemical markers were evaluated. A significant improvement in the biochemical, histopathological, and immunohistochemical parameters were observed in the DM + IRI + BC group, while the DM + IRI + Dex group showed improvements in renal IRI and dyslipidemia. The present study demonstrated that oxidative stress plays a chief role in renal IRI in the STZ-induced diabetic model. Treatment with BC achieved excellent ameliorative effects, while treatment with DEX improved renal IRI.
Keywords:
Diabetes; Dexmedetomidine; Ischemia/Reperfusion; Oxidative Stress
Beneficial Effects of Curcumin Inmaternal and Fetal Oxidativestress and Brain...Prof. Hesham N. Mustafa
This study was planned to explore the protective role of curcumin (Cur) against maternal and fetal oxidative stress and cerebral damage induced by lead (Pb) during pregnancy. Positively pregnant female rats were divided into seven groups: control group, Cur group (300 mg/kg of Cur/b.wt.), DMSO group (50% DMSO), two Pb-treated groups (exposed to 160 and 320 mg/kg b.wt./day of Pb acetate, respectively), and two groups treated with both Pb and Cur (exposed to Pb as previous groups together with 300 mg/kg b.wt./day of Cur). Treatments through oral gavage once a day started from gestation day 1 (GD1) till day 20 (GD20), where the mother rats of different experimental groups were sacrificed to obtain the fetuses. Different chemical parameters were assessed. Brain specimens of mother and fetal groups were processed with examination. The results displayed that Pb administration to pregnant rats resulted in a dose-dependent toxicity for both mothers and fetuses. Also, there was a significant rise in lipid peroxidation and decreased antioxidant enzyme activities in the brains of the different Pb-treated groups. The histological examination of the brain of treated dams and fetuses showed marked alterations. Co-treatment of Cur along with Pb caused a significant decrease in Pb levels as compared with those treated with Pb alone, improving the oxidative condition with amelioration of the brain’s histopathological changes. Co-administration of Cur could have ameliorative effect against Pb-induced neurotoxicity through the reduction of oxidative stress and reversal of histopathological changes.
Keywords:
Lead; Oxidative Stress; Brain; curcumin; Fetal toxicity
Immunohistochemical Study of the Ameliorative Effect of Vitamin E on Liver Re...Prof. Hesham N. Mustafa
The liver is almost unique in its capacity for regeneration after hepatectomy but the exact mechanisms are not yet fully clarified. Antioxidants have been shown to promote liver regeneration after major hepatectomy. The present study evaluated the ameliorative effect of vitamin E administration on the liver regeneration after different periods of partial hepatectomy (PH) in rats. Fifty-six adult male albino rats were divided into three groups: Control sham operated group; partially hepatectomized group which were divided into three subgroups sacrificed at 1day, 3 days and 7days after the operation respectively; Partially Hepatectomized group with vitamin E pretreatment before PH where the rats were given a daily oral dose of vitamin E until the time of sacrifice of the rats. Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and labeling index were demonstrated. After PH, the PCNA positive hepatocytes and the PCNA labeling indices were significantly high after the 1st day and then much decreased after the 3rd day, to be followed by a slight increase at the 7th day. Vitamin E pretreatment in PH rats resulted in a decrease in PCNA positive cells and its labeling indices in the 1st day with a gradual increase in the 3rd and 7th days. Vitamin E has an inhibitory effect in the first 24 hours on liver regeneration followed by stimulatory effect at the third and seventh days after PH. These data indicated that vitamin E pretreatment has an important role in regulation and enhancement of liver regeneration after PH.
Keywords:
Immunohistochemistry; Labeling Index; Vitamin E; Partial Hepatectomy; PCNA
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
2. P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215 207
ing their use in mechanistic studies of SAD and related conditions
[10–12].
Artificial light positive properties at specific illuminance and
exposure time allowed the development of phototherapy for the
treatment of mood and biological disorders associated with sea-
sonal changes. Daily exposure (morning, up to 2-h) to artificial
daylight (bright white artificial light >8000 lx) improved well-
being, sleep, daytime psychomotor vigilance performance, cortisol
and melatonin levels, and SAD patients’ condition [13–18]. Fur-
thermore, strategic exposure to artificial daylight during daytime
improved SAD-like primary and secondary features of Parkinson’s
disease [19], but also learning effectiveness and other cognitive
abilities in people affected by seasonal changes [20,3,21], without
eliciting any major safety concerns [22,23].
Together with other benefits of bright light and the low cost
of the technology, the aforementioned positive effects of pho-
totherapy contribute to the commercial success of daylight-grade
artificial light sources. Surprisingly, considering notably emerg-
ing evidence supporting adverse effects of related electromagnetic
radiations such as UVA [24,25], no report is available on the poten-
tial adverse reactions to long exposure to artificial daylight. In the
present study, we assessed the impact of continuous artificial day-
light exposure during daytime on the physiology, mood, cognitive
functions, and number of cells in the brain of mice.
2. Methods
2.1. Animals and procedures
2.1.1. Animals and light exposure
Swiss female mice (n = 24) were raised under natural daylight,
from birth to the age of 8 months, in the animal facility of the
College of Pharmacy, Qassim University (Buraydah, Saudi Ara-
bia). Then, these animals (36.87 ± 3.83 g weight) were randomly
divided in three groups (n = 8) housed in transparent Plexiglas cages
(70 cm × 70 cm, height 60 cm) in different rooms. A cage was in a
room exposed to natural daylight (22.000 lx average luminance at
cage floor), while the other two were in rooms either exposed to
conventional lighting (500 lx at cage floor) or to artificial daylight
(21.736 lx at cage floor) during the daytime. Lights were switched
on and off by an automated system, based on dawn and dusk. Exper-
iments in live animals were performed in Buraydah, Central Saudi
Arabia, during the winter, thus civilian dawn was at 6:24 a.m. (local
time) and dusk at 6:11 p.m. (daytime length: ∼11h47 min). During
the dark phase (night), rooms were kept under dim red light (3 lx
at cage floor). Light intensity was monitored using a photometer
(9152B, Pasco Scientific, Roseville, CA). In each room the temper-
ature was constant (23.5 ◦C) and animals had ad libitum access to
food and water.
The experiment and all procedures in live animals were
approved by both the Research Center and the Ethical Committee
of the College of Applied Medical Sciences, Qassim University, and
performed according to EC Directive 2010/63/EU on the protection
of animals in scientific experiments.
2.1.2. Experimental procedures
Animals exposed to natural daylight, conventional lighting, or
artificial daylight were monitored daily, to detect signs of systemic
and central nervous system toxicity such as shaggy fur, cachexia,
vocalization when handled, and porphyrin deposits around the eye
(‘red tears’). Negative geotaxis and various reflexes were asserted
(mainly posture, pinna, righting, contact righting, and corneal
reflexes). Animal behavior in cages was recorded with a comput-
erized system equipped with infrared cameras and analyzed to
detect important changes in social behavior. The body weight was
determined every three days.
On exposure days (ED) 14 and 20, mice were submitted to a
battery of behavioral tests aimed at assessing changes in the mood,
cognition, and motor function. Tests were performed between 11
a.m. (Zeitgeber 5, i.e. 5 h after light phase onset) and 2 p.m. (ZT 8),
in rooms where animals were housed. The whole testing proce-
dure required 30 min per animal. The performance of each animal
was video recorded and scored offline. Skin temperature was deter-
mined in both ears at the beginning and the end of the battery of
tests, using a non-contact infrared thermometer.
The experiment in live animals was stopped on ED21 due to
increases in the aforementioned signs of toxicity, in order to com-
ply with animal research ethical standards. Animals were sacrificed
under deep gas anesthesia between ZT 5 and ZT 8. Blood was col-
lected by cardiac puncture and brains dissected out and fixed. Blood
was processed for liver function test. Brains’ left hemispheres were
processed for histopathological studies and stereological estima-
tion of hippocampal neurons at the Department of Histology and
Embryology, Ondokuz Mayis University (Samsun, Turkey). Instead,
the right hemispheres were processed for immunohistochemical
labeling of resident cells and markers of inflammation and apopto-
sis in the entorhinal cortex, at the Department of Anatomy, King
Abdulaziz University (Jeddah, Saudi Arabia).
2.1.3. Artificial daylight exposure
Artificial daylight was delivered by a linear source lamp (LSL)
system designed and optimized to produce isothermal, regular, and
homogeneous electromagnetic radiation by Qassim University’s
Department of Physics [26]. The system was made of a focus-
ing mirror and eight cool white fluorescent tubes (60 cm length,
3.3 cm diameter, 1800 lm) in the same horizontal plane. The dis-
tance between the LSL system and the cage floor was 60 cm.
2.2. Behavioral tests
The following tests were performed sequentially:
2.2.1. Footprint test
The footprint test was performed for gait and balance assess-
ment. Mice with inked paws were allowed to walk freely along
an enclosed box (70 cm long, 7 cm wide, and 20 cm high plexiglas
walls) with a clean sheet of paper placed on the floor. After three
consecutive tests, only one valid trial was considered per animal to
exclude habituation phase-associated abnormal patterns [27].
2.2.2. Elevated plus maze
The EPM consisted of two open arms (30 cm × 7 cm, no wall),
two closed arms (30 cm × 7 cm, with 20 cm high Plexiglas walls),
and a common central platform (7 cm × 7 cm). The entire apparatus
was elevated to 70 cm above floor level. Each mouse was plogical
parameters scored included entries, time spent, and distance trav-
eledlaced on the central platform of the maze, facing an open arm,
and the behavior was recorded for 5 min. Etho in each arm. An entry
occurred when all four limbs were within an arm. Head dips over
the edge of open arms, rearing, grooming, sniffing, and freezing
episodes were also counted.
2.2.3. Open field test
Exploratory behavior was examined in an open field arena in
Plexiglas (40.6 cm × 40.6 cm, height 38.1 cm). The arena’s floor was
divided into a central (20.2 cm × 20.2 cm) and a peripheral zone
(remaining 10.2 cm surrounding the central zone). The test was
started by placing a mouse in the arena, facing the wall. Animal’s
activity in the chamber was recorded for 10 min using a cam-
era mounted on side (approximately 50 cm from the floor) that
3. 208 P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215
captured both vertical and horizontal activities. Mouse locomo-
tor activity was tracked with an application developed in MATLAB
computing environment (MathWorks, Natick, MA) using motion
tracking in image sequences. Entries, time spent, and distance trav-
eled in the central and peripheral zones were determined, as well
as the time spent in the corners.
2.2.4. Y-maze spontaneous alternation test
The test was performed under reduced light (∼100 lx), as recom-
mended [28,29]. Y-maze consisted of three Plexiglas arms (length
30 cm, width 7 cm, height 20 cm) symmetrically placed (120◦ angle
between arms). Each mouse was placed in the center of the maze
and spontaneous activity was recorded for 5 min using a camera
placed above the maze. The following were determined using MAT-
LAB applications: the number and sequence of arm entries, the time
used to perform the first three and the last three alternations, the
total distance traveled, and the average speed (distance traveled
per time unit). Direction changes occurring after sniffing the wall of
another arm (hesitation during arm change) were also determined.
An entry occurred when all four limbs were within the arm.
2.3. Liver function test
Levels of total protein, albumin, globulin, triglycerides and
cholesterol, as well as the activities of the enzymes aspartate
aminotransferase (AST) and alanine aminotransferase (ALT) were
determined in sera using commercially available kits (bio-Merieux
Laboratory Reagents and Products, France), according to the manu-
facturer’s instructions. The albumin/globulins ratio and the fraction
of total protein represented by albumin and globulins were calcu-
lated.
2.4. Tissue processing and histopathological studies
Brains were sequentially fixed in 10% formalin for 8-h, abun-
dantly rinsed in PBS, dried in increasing concentrations of ethanol,
soaked in xylene, and embedded in paraffin using Thermon Shan-
don Citadel tissue processor (GMI Inc., Ramsey, MN, USA).
2.4.1. Histopathology and stereology
Sections of embedded tissues (20 m-thick) were made
throughout the entire left brain hemisphere in the coronal plane,
with a rotary microtome (Leica RM2125RT) using steel blades. After
a random selection of the first section, the rule of systematic ran-
dom sampling with 1/5 ratio was applied to the section selection.
Selected sections were mounted on slides, deparaffinized, rehy-
drated, stained with cresyl violet (Nissl staining), dehydrated, and
then, mounted with Entellan using standard procedures. The mor-
phology of neurons in the hippocampal formation was examined.
Stereological analyses were performed using optical fraction-
ator method [30] and a computerized stereological workstation
equipped with StereoInvestigator software (version 9.0, Micro-
BrieldField; Colchester; USA). Briefly, before starting the counting,
areas (coordinates) of CA1, CA2, CA3, and dentate gyrus were deter-
mined on every section and for each animal using an atlas and a
bright field microscope with motorized stage (microscope objective
4×). These areas of interest were scanned along the x- and y-axes.
The x/y grid sizes, the step size, and the area of unbiased count-
ing frame at microscope objective 100× were determined (i.e. the
most appropriate step and frame to avoid counting CA1–CA3 and
dentate gyrus neurons twice). Afterwards, pyramidal cells (respec-
tively, granular cells) were counted in CA1–CA3 areas (respectively,
the dentate gyrus) using the unbiased counting frame (microscope
objective 100×). Then, total numbers of neurons in the dentate
gyrus and CA1–CA3 were estimated with the optical fractionator
technique (coefficient of error ≤0.05) applied using the relevant
program of the stereological workstation.
2.4.2. Immunohistochemistry
Standard procedures were used, as previously described [31],
on randomly selected sections (6 m-thick) made on paraffin-
embedded brain right hemispheres, in the coronal plane. Primary
antibodies used were goat anti-NeuN (mature neuron marker,
1:1000 dilution), goat anti-GFAP (astrocyte marker, 1:100 dilution),
goat anti-iba1/CD68 (microglial marker, 1:1000 dilution), goat anti-
TNF-␣ (pro-inflammatory cytokine, 1:500 dilution), rabbit anti-Fas
(apoptosis mediator, 1:500 dilution), rabbit anti-Fas ligand (death
receptor, 1:500 dilution), and rabbit anti-caspase 3 (marker of
apoptosis, 1:500 dilution) (Santa Cruz Biotechnology, CA). Biotiny-
lated anti-goat IgG and biotinylated anti-rabbit IgG (1:200 dilution,
Santa Cruz Biotechnology, CA) were the secondary antibodies. The
chromogen substrate 3,3 -diaminobenzidine hydrochloride (DAB)
was applied and counterstained with hematoxylin. Glass coverslips
were placed using DPX mounting medium. Sections labeled were
observed with a computerized light microscope including an Olym-
pus BX53 microscope equipped with an Olympus DP73 camera
(Olympus, Tokyo, Japan), under 4×, 20×, 40× and 100× objectives.
2.5. Data analysis
One way ANOVA for independent groups followed by LSD test
was used to assess the statistical significance of inter-treatment
group changes in body weight, body temperature, and behav-
ioral test parameters. Repeated measures ANOVA followed by LSD
test was used to assess the statistical significance of differences
between ED14 and ED20 performances in behavioral tests. Dif-
ferences with p-value lower than 0.05 were significant. Data are
presented as mean ± SEM.
3. Results
3.1. Animal general condition
3.1.1. Neurological examination
From ED8 onward, artificial daylight–exposed animals were
more agitated when handled than animals of other groups and
expressed audible vocalizations. Besides, fights were more frequent
in the cage of artificial daylight–exposed animals. Thus, three ani-
mals presented with bite injuries on the back on ED10, and almost
all presented with such injuries on legs, feet, and the back on ED20.
These animals also displayed shaggy and dirty furs, decreases in
exploratory activity, regular freezing episodes, as well as slower
visual placing and negative geotaxis responses. They also resisted
more to the separation from grid during the grip strength evalua-
tion.
However, no major locomotor impairment was observed in any
group; grip strength (measured by the ability to hold tight to a grid)
was also comparable between groups; and no marked change was
observed in the reflexes assessed, in the lacrimation (the eyes were
not dry), and in the salivation (no dry mouth).
3.1.2. Body weight and body temperature
The effect of artificial daylight exposure on the body
weight is shown in Fig. 1A. Body weight increased linearly
in, and was comparable between, natural daylight- and con-
ventional lighting–exposed groups (y = 1.02x + 36.1, R2 = 0.97 and
y = 1.17x + 35.6, R2 = 0.98, respectively). However, a comparable
increase was observed in artificial daylight group only between ED
0 and ED 4 (y = 1.75x + 34.88, R2 = 0.99). Afterwards, animals of this
group displayed a transient decrease in body weight following a
polynomial progression (y = 0.58x2 − 3.53x + 41.43, R2 = 0.94), with
4. P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215 209
Fig. 1. Body weight and behavioral tests.
A. Body weight progression. Note the non-linear and lower increase in artificial daylight-exposed animals. B. Body temperature before and after test battery. Note the decrease
in temperature at the end of tests in artificial daylight group on both exposure day (ED) 14 and 20. C–F. Footprints of representative cases of animals exposed to natural
daylight (C) and conventional lighting (D) on ED 20, and animals exposed to artificial daylight on ED 14 (E) and 20 (F). Note the regular pattern of steps in natural daylight
exposed (D), the slightly irregular steps in conventional lighting-exposed (E), and the decreased ground contact surface of hind paws (oval shapes) and incoordination of
walking compared with both natural daylight and conventional lighting groups (arrows) in artificial daylight-exposed (F). G–L. ED 14 (light gray) and EG 20 (dark grey)
behavioral tests’ results. G, H. Distance traveled in the central zone of the open field arena (G) and central zone latency (H). Note the significant decrease in the distance
traveled in the central zone and increase in the latency in artificial light-exposed group. I, J. Time spent in elevated plus maze open arms (I) and closed arm latency (J). Note
the significant decreases compared to natural daylight-exposed group. K, L. Average speed at the first six entries in Y-maze arms (K) and number of arm changes made
with hesitation (L). Note the decrease in speed and increase in arm changes with hesitation in artificial daylight-exposed group more marked on ED 20 than ED 14. Data are
mean ± SEM. ANOVA + LSD test: * P < 0.05, ** P < 0.01, *** P < 0.001.
a restorative inflection point on ED10 (Fig. 1A). The average body
weight of the group exposed to artificial daylight was significantly
lower, compared to the other groups, from ED10 onward (p = 0.034)
(Fig. 1A).
The effect of artificial daylight exposure on pre- and post-battery
test’s body (skin) temperature is shown in Fig. 1B. Body temper-
ature before the battery of behavioral tests was slightly higher in
conventional lighting- and artificial daylight–exposed groups com-
pared to natural daylight–exposed. Body temperature at the end of
the behavioral tests was significantly lower in artificial daylight
group on both ED14 and ED20 compared with values before bat-
tery test (p = 0.03 on ED14, p = 0.01 on ED20) and with post-battery
values of other groups (ED14: p = 0.01 vs. conventional lighting,
p = 0.033 vs. natural daylight/ED20: p = 0.02 vs. conventional light-
ing, p = 0.028 vs. natural daylight) (Fig. 1B).
3.1.3. Serum molecules reflecting hepatic function
The effect of artificial daylight exposure on levels of serum
proteins, lipids, and hepatic enzymes is shown in Table 1. Slight
increases in total cholesterol and triglyceride levels were observed.
Total protein, albumin, and globulin levels were higher in artifi-
cial daylight–exposed group, compared with natural daylight- and,
5. 210 P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215
Table 1
Effect of light source/intensity on levels of serum proteins and molecules reflecting hepatic function.
natural daylight conv. lighting artificial daylight ANOVA: F; p values (F crit = 4.6) Reported rangesa
nat. D vs. Conv. L nat. D vs. art. D Conv. L vs. art. D
Serum protein levels (g/L)
Total protein 70.5 ± 1.2 74 ± 0.5↑
79.6 ± 0.5↑
8; 0.025* 52; 0.0001*** 68;0.0001*** 67 ± 9
Albumin 39.4 ± 1.9 41.3 ± 0.8 47.8 ± 1.4↑
0.9; 0.36 12.4; 0.008** 20; 0.002** 46.5 ± 7.8
% of total 55.9 ± 2.8 55.8 ± 2.8 60 ± 1.7↑
0.001; 1 1.65; 0.235 15.2; 0.005** 38 ± 9.8
Globulin 29.4 ± 1.3 24.5 ± 1.3 38.8 ± 1.5↑
2.4; 0.16 22; 0.0016** 20; 0.002** 29.5 ± 6.4
% of total 41.5 ± 6 33 ± 3.7 48.8 ± 1.7↑ 4.5; 0.06 10; 0.012* 15; 0.005** 38 ± 9.8
A/G ratio 1.4 ± 0.26 1.8 ± 0.53 1.2 ± 0.06 0.14; 2.711 0.31; 1.13 0.047; 5.49* 1.7 ± 0.34
Hepatic enzymes and lipid levels (mg/dL)
ALT (IU/L) 68.7 ± 39.2 71.5 ± 36.4 186.6 ± 63.1 0.91; 0.013 12.6; 0.008** 12.5;0.007** 60 ± 13.2
AST (IU/L) 29 ± 13.5 39.6 ± 7.6 67.4 ± 27 0.16; 2.315 8.11; 0.021* 0.05; 5.96 38 ± 21.9
T. cholesterol 125 ± 60 121 ± 60 118 ± 80 0.1; 0.81 0.001; 0.97 0.04; 0.85 110 ± 29
Triglycerides 231 ± 63 225 ± 63 248 ± 31 0.01; 0.9 0.06; 0.81 0.5; 0.51 150 ± 42
a
Ranges reported by recent studies performed under normal laboratory lighting in mice [33,20,37,3]. T. cholesterol: total cholesterol. ANOVA +LSD test: *P < 0.05; **P < 0.01;
***P < 0.001. Data are mean ± SEM.
in a lesser extent, conventional lighting–exposed. The levels of
these proteins were also higher than physiological values [32–34]
(Table 1). Albumin/globulins ratio in artificial daylight group was
significantly lower (respectively, slightly lower) compared to con-
ventional lighting (respectively, natural daylight).
Serum levels of the liver enzymes aspartate aminotransferase
(AST) and alanine aminotransferase (ALT) were higher in artifi-
cial daylight–exposed animals compared to natural daylight- and
conventional lighting–exposed. Levels of AST and ALT in artificial
light group were higher than physiological values reported in the
literature [32–34] (Table 1).
3.2. Behavioral tests
3.2.1. Footprints’ analysis
Representative cases of animals exposed to natural daylight,
conventional lighting, and artificial daylight are shown in Fig. 1C–F.
Artificial daylight–exposed animals displayed static gait alterations
marked by a decreased size of contact (more obvious in hind paws)
and an incoordination of walking (gait ataxia) on ED20 (Fig. 1F)
compared with their walking performance on ED14 (Fig. 1E), and
also compared with both natural daylight- (Fig. 1C) and conven-
tional lighting–exposed (Fig. 1B). Notably, some animals exposed
to conventional lighting presented slightly irregular step patterns
compared with animals exposed to natural daylight.
3.2.2. Open field
Open field test results are shown in Fig. 1G, H and Table 2. The
distance traveled in the open field arena was significantlydecreased
in artificial daylight–exposed group on both ED14 (p = 0.002)
and ED20 (p = 0.0059) compared with natural daylight–exposed
(Table 2). The distance traveled in the central zone of the arena
was significantly lower in conventional lighting- and artificial
daylight–exposed compared with natural daylight–exposed on
both ED14 (p = 0.014 and p = 0.009, respectively) and ED20 (p = 0.03
and p = 0.009, respectively). The distance traveled in the central
zone by artificial daylight–exposed was also significantly lower
than conventional lighting–exposed (p = 0.048) (Fig. 1G). Time to
the first entry in the central zone (central zone latency) was
significantly higher in artificial daylight–exposed group on ED20
compared with ED14 (p = 0.042), but compared also to the other
groups (p = 0.014 vs. natural daylight– and p = 0.03 vs conventional
lighting–exposed) (Fig. 1H). The time spent in corners was signif-
icantly higher in artificial daylight–exposed compared with the
other groups on ED14 (p = 0.0013 vs. natural daylight, p = 0.0034
vs conventional lighting) and ED20 (p = 0.005 vs. natural daylight,
p = 0.039 vs conventional lighting) (Table 2). Central zone entries
and relative time, rearing (on hind legs and against wall), and
grooming latency were significantly decreased in artificial day-
light group, while freezing and sniffing episodes were increased
(Table 2).
3.2.3. Elevated plus maze
The results of EPM test are shown in Fig. 1I, J and Table 2.
The time spent in maze open arms was significantly decreased
in artificial daylight–exposed group compared with conventional
lighting–exposed (p = 0.001 on ED14 and p = 0.0012 on ED20) and
natural daylight–exposed (p = 0.0051 on ED14 and p = 0.0029 on
ED20) (Fig. 1I). Closed arm latency was significantly decreased
in artificial daylight–exposed group compared with conventional
lighting– (p = 0.052 on ED14 and p = 0.037 on ED20) and natural
daylight–exposed (p = 0.0051 on ED14 and p = 0.0023 on ED20).
Closed arm latency of conventional lighting–exposed was also
shorter compared with natural daylight–exposed (p = 0.037 on
ED20) (Fig. 1J).
Rearing against wall frequency was significantly increased in
conventional lighting- (p = 0.018) and in artificial daylight–exposed
(p = 0.002) on ED14 compared with natural daylight. Instead,
on ED20 a decrease in this parameter was observed in artifi-
cial daylight–exposed compared with ED14 (p = 0.0001), but also
conventional lighting– (p = 0.025) and natural daylight–exposed
(p = 0.007) (Table 2). As also shown in Table 2, open arm entries,
central platform time, and head dips over the open arms, and
freezing episode frequency were significantly decreased in artifi-
cial daylight–exposed group compared to natural daylight, and in
a lesser extent, conventional lighting. On the other hand, fecal boli
and episodes of rearing, undirected sniffing, and grooming attempts
(grooming episodes shorter than 3 s) were increased (Table 2).
3.2.4. Y-maze
Y-maze test results are shown in Fig. 1K, L and Table 2. The
average speed in the first six entries was decreased in artificial
daylight–exposed group on ED20 compared with ED14 (p = 0.008)
and to conventional lighting–exposed (p = 0.001), but not natu-
ral daylight–exposed (Fig. 1K). A similar scenario was observed
with the speed difference between the first six and the last six
entries (Table 2). The frequency of episodes of arm change with
hesitation (sniffing of an arm entrance followed by the choice of
another arm) was significantly higher in artificial daylight–exposed
group on ED20 compared with ED14 (p = 0.003) and to other groups
(p = 0.0003 vs. conventional lighting–exposed, p = 0.0001 vs. natural
daylight–exposed) (Fig. 1L). Arm change with hesitation episodes
was more common in conventional lighting–exposed than in nat-
ural daylight–exposed (p = 0.01) (Fig. 1L).
6. P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215 211
Table 2
Effect of light exposure on animal performance in behavioral tests.
Natural daylight Conventional lighting Artificial daylight
ED14 ED20 ED14 ED20 ED14 ED20
Open field
Total distance, cm 2930 ± 250 2561 ± 200 2072 ± 197 b↓
2159 ± 257 1801 ± 190 b↓
1607 ± 217 b↓
Central zone time, % total 13.4 ± 2.1 12.7 ± 2.2 18 ± 3 18.5 ± 3.4 11.2 ± 2.1c↓
5.9 ± 1.4a,b,c↓
Central zone entries, nr 22 ± 1.8 21.8 ± 2 15.1 ± 2.6b↓
16.1 ± 2.3b↓
13.7 ± 1.7b↓
11.9 ± 2.9b↓
Time spent in corners, s 119.1 ± 26 111.6 ± 28 156.8 ± 22 174.3 ± 29 257.3 ± 26a,b↑
246.9 ± 31 a,b↑
Rearing on hind legs, nr 26.2 ± 5.7 25.6 ± 6.3 32.6 ± 6.3 35.7 ± 6.8 38.1 ± 13.8 18.4 ± 5.5c↓
Rearing against wall, nr 41.1 ± 3 39.8 ± 2.5 51 ± 5.5 55.3 ± 4.9b↑
51.3 ± 8.3 40.7 ± 6c↓
Undirected sniffing, nr 24 ± 3 21.4 ± 1.6 34.9 ± 4.4b↑
38.4 ± 5.3b↑
51.3 ± 8.7b↑
62.6 ± 3.3b,c↑
Wall sniffing, nr 34.1 ± 3.1 34.6 ± 2.9 48.1 ± 4b↑
52.7 ± 4b↑
47.4 ± 6.4b↑
77.3 ± 5.9a,b,c↑
Floor sniffing, nr 13.1 ± 1.3 12.2 ± 1.1 20.8 ± 2.3b↑
23.1 ± 3b↑
41.9 ± 3.1b,c↑
67.3 ± 4.5a,b,c↑
Grooming, nr 5.1 ± 1.4 5.8 ± 1.4 7.3 ± 1.5 7.3 ± 1.6 6.6 ± 1.5 5.4 ± 1.2
Grooming latency, s 191.8 ± 44 193 ± 38.5 85.8 ± 23b↓
92 ± 23.4b↓
84.1 ± 4.3b↓
99.9 ± 15b↓
Freezing episodes, nr 1.5 ± 1 1.4 ± 0.7 1.6 ± 0.5 1.3 ± 0.5 4.1 ± 1.5 4.1 ± 0.9b,c↑
Elevated plus maze
Distance, cm 411 ± 60 420 ± 65 544 ± 61 527 ± 58 469 ± 45 390 ± 55
Total arm entries, number 13.1 ± 2.8 13.4 ± 3.1 15.3 ± 2.7 15.1 ± 1.7 13.1 ± 1.9 7.9 ± 2.0a,c↓
Open arm entries, % total 51.6 ± 8.9 50.6 ± 9.9 51.8 ± 5.4 50.9 ± 5.6 17 ± 2.7b,c↓
19.5 ± 4.9b,c↓
Central platform time, s 21.9 ± 2.4 22.2 ± 2.6 32.3 ± 5.3 25.9 ± 5.1 31.1 ± 5.2c↓
18.4 ± 4.3c↓
Head dips over open arm, nr 18.1 ± 2.5 18.6 ± 2.7 37.3 ± 5b↑
34.3 ± 6.4b↑
15.3 ± 1.9c↓
13.9 ± 2.7c↓
Rearing against wall, nr 5.7 ± 2.0 6.2 ± 2.0 13.5 ± 2.1b↑
14.4 ± 1.9b↑
17.4 ± 2.0 b↑
7.1 ± 1.2a,c↓
Rearing, nr 3.5 ± 1.7 4 ± 1.8 14.4 ± 4b↑
14.1 ± 3.1b↑
14.4 ± 3.4b↑
11.3 ± 2.6a,c↓ b↑
Sniffing, nr 18.2 ± 2.2 18.2 ± 2.5 40.9 ± 3.8b↑
44.6 ± 4.6b↑
63.4 ± 8.1b,c↑
62.3 ± 5.7b,c↑
Grooming attempts, nr 5.6 ± 1.0 5.8 ± 1.1 4.6 ± 1.1 5.8 ± 0.9 3.7 ± 0.6 6.6 ± 0.8a↑
Freezing, nr 2.7 ± 1.1 2.8 ± 1.1 2.1 ± 0.5 1.8 ± 0.7 3.6 ± 1.0 4.3 ± 0.6c↑
Fecal boli, nr 1.2 ± 0.3 0.8 ± 0.3 1.3 ± 0.5 1.9 ± 0.6 1.9 ± 0.9 2.3 ± 0.7b↑
Y maze
Distance, cm 938 ± 82 818 ± 98 1330 ± 168b↑
1320 ± 188b↑
1349 ± 137b↑
1181 ± 93b↑
Alternation, nr 19.2 ± 1.4 18.8 ± 1.6 26.6 ± 2.7b↑
27.8 ± 3.7b↑
24.3 ± 3.1 18.6 ± 1.8c↓
X-Y-Z alternations, nr 10.5 ± 0.8 10 ± 0.6 15.4 ± 1.7b↑
17.2 ± 2.4b↑
15.0 ± 2.5 10.9 ± 1.4c↓
Speed first entries – last, cm/s 3.6 ± 1.1 3.5 ± 1.2 6.1 ± 0.4 6.0 ± 0.7 6.6 ± 0.9 4.0 ± 0.5a,c↓
ED14, ED20: exposure days 14 and 20. a,b,c: ANOVA +LSD test, vs. respective exposure day 14 (a), vs. natural daylight (b), and vs. conventional lighting (c). Data are mean ± SEM.
The distance traveled in the Y-maze was significantly shorter
in natural daylight–exposed than in other groups on both ED14
(p = 0.012 vs. conventional lighting, p = 0.001 vs. artificial day-
light) and ED20 (p = 0.01 vs. conventional lighting, p = 0.004 vs.
artificial daylight) (Table 2). On the same hand, the frequency
of consecutive visits of three different arms was higher in con-
ventional lighting–exposed group on ED14 (p = 0.008) and ED20
(p = 0.005) compared with natural –exposed, but on ED20 artifi-
cial daylight–exposed displayed a lower frequency compared with
conventional lighting–exposed (p = 0.03). Similar, arm alternation
frequency was also lower in natural daylight–exposed, except on
ED20 where a drop was observed in artificial daylight–exposed
(p = 0.02 vs. conventional lighting).
3.3. Brain histopathological studies
3.3.1. Neuronal morphology and counts
The analysis of Nissl-stained neurons in the hippocampal forma-
tion revealed marked neuronal loss in artificial daylight–exposed
animals. This loss was particularly marked in the dentate gyrus.
Fig. 2A–C shows dentate gyrus granule cells of representative
cases of animals exposed to natural daylight, conventional light-
ing, and artificial daylight. While dentate gyrus granule cells had
normal shapes in natural daylight- (Fig. 2A) and conventional
lighting–exposed (Fig. 2B) animals, these neurons were condensed
in artificial daylight–exposed (Fig. 2C).
Results of neuronal counts in CA1, CA2, and CA3, and in the
dentate gyrus are shown in Fig. 2D, E. Absolute counts of pyrami-
dal neurons in CA1, CA2, and CA3 revealed significant decreases
in artificial daylight–exposed (P < 0.001), and in a lesser extent,
in conventional lighting–exposed animals compared with natu-
ral daylight–exposed (Fig. 2D). Similar results were obtained in
counts of dentate gyrus granule cells (Fig. 2D). Relative values of
neuronal counts (percent of natural daylight counts) suggested
that the dentate gyrus was more affected than other structures
of the hippocampal formation in artificial daylight–exposed com-
pared to conventional lighting–exposed (Fig. 2E). Relative counts
also revealed a decrease (∼15%) in pyramidal neurons in conven-
tional lighting–exposed animals (but not in granule cells) compared
with natural daylight–exposed (Fig. 2E).
3.3.2. Expressions of inflammation, apoptosis, and brain resident
cell markers
The expressions of markers labeled in the entorhinal cortex
of representative cases of animals exposed to natural daylight,
conventional lighting, and artificial daylight are shown in Fig. 3.
The markers of activated microglia (iba-1) (Fig. 3A–C), astrocytes
(GFAP) (Fig. 3D–F), and neurons (NeuN) (Fig. 3G–I) were overex-
pressed in most artificial daylight–exposed animals, compared to
the other groups. In addition, the first displayed enlarged astro-
cytes, microglia in activated macrophage shape, and dead (or dying)
neurons (Fig. 3A–I).
Also unlike the other groups, animals of the artificial
daylight–exposed group displayed increased expressions of: (i)
the pro-inflammatory cytokine TNF-␣ (Fig. 3J–L); (ii) the receptor
mediating cell-cell interaction-induced cell death associated with
marked inflammation FAS (Fig. 3M–O) and its ligand (Fig. 3P–R);
and (iii) the marker of apoptosis caspase 3 (Fig. 3S–U).
4. Discussion
The results of the present study suggest systemic and central
nervous system functional alterations in mice exposed exclusively
to artificial daylight during daytime for 20 days. Notably, signif-
icant decreases in the frequency of rearing against wall, i.e. in
attempts to escape, in the open field and the elevated plus maze
7. 212 P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215
Fig. 2. Hippocampal neuron observation and counts.
A–C. Micrographs of Nissl-stained dentate gyrus neurons of representative cases of animals exposed to natural daylight (A), conventional lighting (B), and artificial daylight
(C). Note the dead (black arrows) and the condensed cells (white arrows) in arficial daylight-exposed (C) and the normal shapes of granular cells in the other groups (asterisks)
(A, B). D, E. Hippocampal neuron counts. Absolute (D) and relative (E) cell counts in the CA1–CA3 (pyramidal cells) and dentate gyrus (DG, granular cells) of animals exposed
to natural daylight, conventional lighting, or artificial daylight during daytime for 20 days. Note the marked decreases in neuronal population in artificial daylight-exposed
group. Data are mean ± SEM.
(EPM), as well as drastic decreases in the speed at first entries in
Y-maze arms were observed in artificial daylight group on ED20,
unlike ED14 and unlike natural daylight and conventional light-
ing groups. These observations indicate that animals exposed to
artificial daylight developed coping deficit for the aversive situa-
tions presented by the ethological tests [35,36]. Other indicators of
depression were observed in artificial daylight–exposed animals in
the third week, including: shaggy and dirty fur, decreased groom-
ing, increased aggressiveness, regular freezing episodes, decreased
exploratory activity, and body weight loss.
The performance of artificial daylight–exposed animals in the
behavioral tests performed on ED20 also suggested other signs
of cognitive and motor impairment. Anxiety was suggested by
increases in sniffing episodes and by post-testing hypothermia,
a hallmark of mild chronic stress exposure [37,38]. In addition,
increased latency to first entry in the anxiogenic areas of the
EPM (open arms) and of the open field arena (central zone)
were observed, together with decreases in the distance trav-
eled in these areas. Moreover, thigmotaxis, a stable and robust
indicator of discomfort manifested by the tendency to remain
close to the walls (secure area) [39,40] was markedly increased
in artificial daylight–exposed, as indicated by increases in the
number of activities close to walls in all the behavioral tests per-
formed. Besides, a shift in behavioral baseline was indicated by
increases in EPM central platform time [41]. Prefrontal cortex
dysfunction was indicated by decreased spontaneous alternations
in the Y-maze [42,43]. Spatial memory impairment was indi-
cated by increases in episodes of arm change with hesitation and
decreases in consecutive explorations of three different arms in the
Y maze [42,44,45]. Furthermore, the incoordination of walking and
decreased ground contact surface of hind paws observed in artifi-
cial daylight-exposed, compared with the other groups, indicated
a motor dysfunction with central nervous system involvement.
We investigated the affection of a major sensorimotor gating
structure, the entorhinal cortex, as a potential mechanism for the
cognitive deficits induced by the exposure to artificial daylight.
Immunohistochemical labeling of resident cells in the entorhinal
cortex of artificial daylight–exposed animals revealed the pres-
ence of enlarged astrocytes and brain macrophages, suggesting
that astrocytes and microglia were activated. On the same hand,
the pro-inflammatory cytokine TNF-␣ was overexpressed, and
many resident cells were positive for the inflammation-induced
death receptor Fas and its ligand. Neuronal marker NeuN was
overexpressed, indicating that neurons were injured, and many
neurons were positive for the marker of apoptosis caspase 3. These
observations suggested the presence of a marked inflammation
in the entorhinal cortex associated with neuronal loss. Using ste-
reological techniques, we estimated the number of Nissl-stained
neurons in the hippocampal formation, a major functional tar-
get of the entorhinal cortex. Losses in dentate gyrus granule cells
and as well as CA1–CA3 pyramidal neurons of hippocampus were
observed, and cell numbers were significantly decreased. These
findings suggest that neuronal loss in the neocortex-entorhinal
cortex-hippocampus axis resulted at least partly from detrimental
neuroinflammation, and were among the drivers of brain functional
alterations observed in this study following replacement of natural
daylight with artificial daylight.
Furthermore, liver function test, performed to assess general
health status, suggested an affection of the hepatic function in arti-
ficial daylight–exposed animals. Marked increases in serum levels
8. P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215 213
Fig. 3. Entorhinal cortex immunolabeling.
Representative cases of natural daylight, conventional lighting, and artificial daylight-exposed animals. A–I. Brain resident cells: microglia (iba-1) (A–C), astrocytes (GFAP)
(D–F), and neurons (NeuN) (G–I). J–U. Markers of inflammation and apoptosis: TNF-␣ (J–L), Fas (M–O), Fas ligand (P–R), and caspase 3 (S–U). Note the dying neurons in the
artificial daylight-exposed (arrows in I, U), the increased expressions of TNF-␣ (L), Fas (O), Fas ligand (R), and caspase 3 (U), as well as the marked activation of microglia (C)
and astrocytes (F).
of the enzymes AST and ALT were observed, as well as decreased
albumin/globulins ratio, and abnormally high levels of total pro-
teins, albumin, and globulins. Considering that these alterations are
well-established indicators of high-grade hepatotoxicity [46–48], it
appears that artificial daylight would have resulted in animal death
if the exposure had continued. This finding suggests that liver-to-
brain signaling may also have accounted among the causative or
aggravating factors of the cognitive deficits and mood alterations
observed in the present study [49–51].
Altogether, our results suggest that long hour exposure to arti-
ficial daylight may have tremendous negative effects on mouse
brain. Interestingly, an early study in humans reported mild cog-
nitive impairment following daytime exposure to bright artificial
light [52]. Thus, artificial daylight exposure for long hours may
be detrimental for mental health in humans as well. Alarmingly,
mouse exposure to conventional lighting in our study induced a
mild cognitive deficit and motor impairment. Although this obser-
vation should be verified in humans, it raises caution over keeping
children indoor under conventional lighting during the daytime.
For instance, teaching institutions should probably avoid using con-
ventional lighting when natural daylight is available.
5. Conclusions
The present study addressed the consequences of replacing nat-
ural daylight by either artificial daylight or conventional lighting for
general health and cognition in mice. Mice presented with signs of
hepatotoxicity, as well as increasing signs of neurotoxicity, anxi-
ety, depression, cognitive alterations, and motor impairment that
became severe in the third week of exposure. The involvement of
the neocortex-entorhinal cortex-hippocampus axis was suggested
by immunohistochemical studies that revealed neuroinflammation
and neuronal loss in the entorhinal cortex, but also by massive neu-
ronal loss in the hippocampal formation confirmed by stereological
cell estimation. Mild cognitive and motor impairments were also
observed in animals exposed to conventional lighting. These find-
ings suggest that the replacement of natural daylight by artificial
daylight, and in a lesser extent conventional lighting, had detri-
9. 214 P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215
mental effects on the brain function and general health in mice.
Our findings raise concern over the extended use of artificial light
during the daytime, and call for studies assessing the intensity and
exposure time safe for humans, particularly for children, the most
sensible group in the human population.
Conflicts of interest statement
Authors declare no conflict of interest.
Acknowledgements
The present study was supported by the College of Applied Med-
ical Sciences, Qassim University (Saudi Arabia), the Department of
Histology and Embryology, Medical Faculty of Ondokuz Mayis Uni-
versity, (Turkey), and the Department of Anatomy, King Abdulaziz
University (Saudi Arabia). Authors acknowledge the technical sup-
port of Prof. Osama H. Omer, and of the Colleges of Physics and
Pharmacy, Qassim University.
References
[1] Allebrandt KV, Teder-Laving M, Kantermann T, Peters A, Campbell H, Rudan I,
et al. Chronotype and sleep duration: the influence of season of assessment.
Chronobiol Int 2014;31:731–40.
[2] Arendt J. Biological rhythms during residence in polar regions. Chronobiol Int
2012;29:379–94.
[3] Munch M, Linhart F, Borisuit A, Jaeggi SM, Scartezzini JL. Effects of prior light
exposure on early evening performance, subjective sleepiness, and hormonal
secretion. Behav Neurosci 2012;126:196–203.
[4] Roenneberg T, Kantermann T, Juda M, Vetter C, Allebrandt KV. Light and the
human circadian clock. Handb Exp Pharmacol 2013:311–31.
[5] Turner PL, Mainster MA. Circadian photoreception: ageing and the eye’s
important role in systemic health. Br J Ophthalmol 2008;92:1439–44.
[6] Belleville G, Foldes-Busque G, Dixon M, Marquis-Pelletier E, Barbeau S, Poitras
J, et al. Impact of seasonal and lunar cycles on psychological symptoms in the
ED: an empirical investigation of widely spread beliefs. Gen Hosp Psychiatry
2013;35:192–4.
[7] Charlton BG. A model for self-treatment of four sub-types of symptomatic
‘depression’ using non-prescription agents: neuroticism (anxiety and
emotional instability); malaise (fatigue and painful symptoms); demotivation
(anhedonia) and seasonal affective disorder ‘SAD’. Med Hypotheses
2009;72:1–7.
[8] Lawrynowicz AE, Baker TD. Suicide and latitude in Argentina: Durkheim
upside-down. Am J Psychiatry 2005;162:1022.
[9] Postolache TT, Lapidus M, Sander ER, Langenberg P, Hamilton RG, Soriano JJ,
et al. Changes in allergy symptoms and depression scores are positively
correlated in patients with recurrent mood disorders exposed to seasonal
peaks in aeroallergens. Sci World J 2007;7:1968–77.
[10] Ebling FJ. On the value of seasonal mammals for identifying mechanisms
underlying the control of food intake and body weight. Horm Behav
2014;66:56–65.
[11] Ferguson SA, Maier KL. A review of seasonal/circannual effects of laboratory
rodent behavior. Physiol Behav 2013;119:130–6.
[12] Rendon NM, Rudolph LM, Sengelaub DR, Demas GE. The agonistic adrenal:
melatonin elicits female aggression via regulation of adrenal androgens. Proc
Biol Sci 2015:282.
[13] Gabel V, Maire M, Reichert CF, Chellappa SL, Schmidt C, Hommes V, et al.
Effects of artificial dawn and morning blue light on daytime cognitive
performance, well-being, cortisol and melatonin levels. Chronobiol Int
2013;30:988–97.
[14] Lewy AJ. Circadian misalignment in mood disturbances. Curr Psychiatry Rep
2009;11:459–65.
[15] Phipps-Nelson J, Redman JR, Dijk DJ, Rajaratnam SM. Daytime exposure to
bright light, as compared to dim light, decreases sleepiness and improves
psychomotor vigilance performance. Sleep 2003;26:695–700.
[16] Rojas JC, Bruchey AK, Gonzalez-Lima F. Low-level light therapy improves
cortical metabolic capacity and memory retention. J Alzheimers Dis
2012;32:741–52.
[17] Slusher TM, Vreman HJ, Olusanya BO, Wong RJ, Brearley AM, Vaucher YE, et al.
Safety and efficacy of filtered sunlight in treatment of jaundice in African
neonates. Pediatrics 2014;133:e1568–74.
[18] Yoshida A, Takagi A, Ikejima A, Takenaka H, Fukai T, Ikeda S. A retrospective
study of 231 Japanese vitiligo patients with special reference to phototherapy.
Acta Dermatovenerol Croat 2014;22:13–8.
[19] Willis GL, Turner EJ. Primary and secondary features of Parkinson’s disease
improve with strategic exposure to bright light: a case series study.
Chronobiol Int 2007;24:521–37.
[20] Mott MS, Robinson DH, Williams-Black TH, McClelland SS. The supporting
effects of high luminous conditions on grade 3 oral reading fluency scores.
Springerplus 2014;3:53.
[21] Wahnschaffe A, Haedel S, Rodenbeck A, Stoll C, Rudolph H, Kozakov R, et al.
Out of the lab and into the bathroom: evening short-term exposure to
conventional light suppresses melatonin and increases alertness perception.
Int J Mol Sci 2013;14:2573–89.
[22] Howland RH. Somatic therapies for seasonal affective disorder. J Psychosoc
Nurs Ment Health Serv 2009;47:17–20.
[23] Lane KL, Hovenic W, Ball K, Zachary CB. Daylight photodynamic therapy: the
Southern California experience. Lasers Surg Med 2015;47:168–72.
[24] Lerche CM, Heerfordt IM, Heydenreich J, Wulf HC. Alternatives to outdoor
daylight illumination for photodynamic therapy–use of greenhouses and
artificial light sources. Int J Mol Sci 2016;17:309.
[25] Richer V, Lui H. Cross-sectional evaluation of acute adverse reactions during
UVA1 phototherapy. Br J Dermatol 2017, http://dx.doi.org/10.1111/bjd.15319.
[26] Abou-Zaid FA, Omar KHM, Salim EI, El-Deen AA, Bekhite MM. Can man made
visible light radiation affect the reproductive capacity of male mice? Egypt J.
Exp. Biol 2006;2:81–91.
[27] Vandeputte C, Taymans JM, Casteels C, Coun F, Ni Y, Van LK, et al. Automated
quantitative gait analysis in animal models of movement disorders. BMC
Neurosci 2010;11:92.
[28] Arendash GW, Gordon MN, Diamond DM, Austin LA, Hatcher JM, Jantzen P,
et al. Behavioral assessment of Alzheimer’s transgenic mice following
long-term Abeta vaccination: task specificity and correlations between Abeta
deposition and spatial memory. DNA Cell Biol 2001;20:737–44.
[29] Kulesskaya N, Voikar V, Peltola M, Yegutkin GG, Salmi M, Jalkanen S, et al.
CD73 is a major regulator of adenosinergic signalling in mouse brain. PLoS
One 2013;8:e66896.
[30] West MJ, Slomianka L, Gundersen HJ. Unbiased stereological estimation of the
total number of neurons in thesubdivisions of the rat hippocampus using the
optical fractionator. Anat Rec 1991;231(December (4)):482–97.
[31] Farahna M, Seke Etet PF, Osman SY, Yurt KK, Amir N, Vecchio L, et al. Garcinia
kola aqueous suspension prevents cerebellar neurodegeneration in long-term
diabetic rat – a type 1 diabetes mellitus model. J Ethnopharmacol
2017;195:159–65.
[32] Habib MR, Karim MR. Effect of anhydrosophoradiol-3-acetate of Calotropis
gigantea (Linn.) flower as antitumoric agent against Ehrlich’s ascites
carcinoma in mice. Pharmacol Rep 2013;65:761–7.
[33] Mostafavi SH, Fazilati M, Mostafavi SA, Vahhabi MR, Mostafavi F, Omidvarinia
S, et al. Hepatotoxicity of Dorema aucheri (Bilhar) in albino mice. Arch Iran
Med 2013;16:530–2.
[34] Perez Gutierrez RM, Madrigales AD, Horcacitas MC, Garcia BE, Cruz VT,
Mota-Flores JM. Ameliorative effect of hexane extract of phalaris canariensis
on high fat diet-induced obese and streptozotocin-induced diabetic mice.
Evid Based Complement Altern Med 2014;2014:145901.
[35] Chourbaji S, Zacher C, Sanchis-Segura C, Dormann C, Vollmayr B, Gass P.
Learned helplessness: validity and reliability of depressive-like states in mice.
Brain Res Brain Res Protoc 2005;16:70–8.
[36] Overmier JB, Seligman ME. Effects of inescapable shock upon subsequent
escape and avoidance responding. J Comp Physiol Psychol 1967;63:28–33.
[37] Mrdalj J, Lundegaard MA, Murison R, Konow JF, Milde AM, Pallesen S, et al.
Hypothermia after chronic mild stress exposure in rats with a history of
postnatal maternal separations. Chronobiol Int 2014;31:252–64.
[38] Richter SH, Wollmann E, Schmidt M, Zillmann U, Hellweg R, Sprengel R, et al.
The effects of neonatal cryoanaesthesia-induced hypothermia on adult
emotional behaviour and stress markers in C57BL/6 mice. Behav Brain Res
2014;270:300–6.
[39] Blaszczyk JW, Tajchert K, Lapo I, Sadowski B. Acoustic startle and open-field
behavior in mice bred for magnitude of swim analgesia. Physiol Behav
2000;70:471–6.
[40] Leppanen PK, Ewalds-Kvist SB, Selander RK. Mice selectively bred for
open-field thigmotaxis: life span and stability of the selection trait. J Gen
Psychol 2005;132:187–204.
[41] Rodgers RJ, Lee C, Shepherd JK. Effects of diazepam on behavioural and
antinociceptive responses to the elevated plus-maze in male mice depend
upon treatment regimen and prior maze experience. Psychopharmacology
(Berl) 1992;106:102–10.
[42] Gotesson J, Ericson M, Soderpalm B, Pickering C. Repeated ethanol but not
phencyclidine impairs spontaneous alternation behaviour in the Y-maze.
Basic Clin Pharmacol Toxicol 2012;110:347–52.
[43] Hu R, Gong X, Duan Y, Li N, Che Y, Cui Y, et al. Neurotoxicological effects and
the impairment of spatial recognition memory in mice caused by exposure to
TiO2 nanoparticles. Biomaterials 2010;31:8043–50.
[44] Harquin Simplice F, David ET, Herve Herve NA. Enhancing spatial memory:
anxiolytic and antidepressant effects of tapinanthus dodoneifolius (DC)
danser in mice. Neurol Res Int 2014;2014:974308.
[45] Rusu G, Popa G, Ochiuz L, Nechifor M, Tartau L. Effects of some dopamine
antagonists on spatial memory performance in rats?experimental research.
Rev Med Chir Soc Med Nat Iasi 2014;118:116–24.
[46] Du XJ, Tang LL, Mao YP, Sun Y, Zeng MS, Kang TB, et al. The pretreatment
albumin to globulin ratio has predictive value for long-term mortality in
nasopharyngeal carcinoma. PLoS One 2014;9:e94473.
[47] Jolles S, Borrell R, Zouwail S, Heaps A, Sharp H, Moody M, et al. Calculated
globulin (CG) as a screening test for antibody deficiency. Clin Exp Immunol
2014;177:671–8.
10. P.F. Seke Etet et al. / Journal of Microscopy and Ultrastructure 5 (2017) 206–215 215
[48] Mwanza T, Miyamoto T, Okumura M, Kadosawa T, Fujinaga T.
Ultrasonographic evaluation of portal vein hemodynamics in experimentally
bile duct ligated dogs. Jpn J Vet Res 1998;45:199–206.
[49] D’Mello C, Swain MG. Liver-brain interactions in inflammatory liver diseases:
implications for fatigue and mood disorders. Brain Behav Immun
2014;35:9–20.
[50] Datar S, Wijdicks EF. Neurologic manifestations of acute liver failure. Handb
Clin Neurol 2014;120:645–59.
[51] Shawcross DL, Wendon JA. The neurological manifestations of acute liver
failure. Neurochem Int 2012;60:662–71.
[52] Lafrance C, Dumont M, Lesperance P, Lambert C. Daytime vigilance after
morning bright light exposure in volunteers subjected to sleep restriction.
Physiol Behav 1998;63:803–10.