Studying the expression of proteins in Chrysichthys nigrodigitatus muscle, is essential to understand the biological, physiological and ecological aspects that may be of advantage in ecotoxicology as a tool for biomonitoring the effects of environmental pollution, as well as food safety. This study was aimed to accomplish a systematic characterization of the muscle proteome as well as to identify a putative set of protein biomarkers in C. nigrodigitatus to environmental pollution in Ologe and Badagry lagoons. Fifteen fi sh samples were used
as representative of the population for proteomics analysis. 116 proteins was expressed with 70 up-regulated, 25 down-regulated from Ologe Lagoon and 30 up-regulated, 17 down-regulated proteins from Badagry Lagoon expressed in the muscle of C. nigrodigitatus in response to environmental stressors using iTRAQ, while 8 protein spots from 2-D gels, representing 8 proteins with 2 up-regulated and 6 down-regulated, have been identifi ed using MALDI TOF/TOF MS. Pearson correlation revealed signifi cant correlation (p < 0.05) betwee environmental variables and protein markers. The investigation revealed that the expressed proteins in the muscle of C. nigrodigitatus served as a prognostic tool to assess the fi sh health and pollution status, which was observed that the fi sh were physiologically perturbed by environmental stressors in Ologe and Badagry lagoons.
Foodomics - the application of advanced omics technologies to understand the molecular and genetics level in food and correleate with nutrition and authenticationn purposes. (getnet)
Metagenomics to Unlock the Biotechnological Potential of Marine Environments by Michele de Cássia Pereira e Silva in Examines in Marine C Biology & Oceanography
Nutrients present in Nutrease powder play an important role in maintaining the normal functions of the human body. The major nutrients present in Nutrease powder include Natural carbohydrates, proteins, lipids, vitamins, and minerals. Besides these, there are some bioactive food components known as “phytonutrients” that play an important role in human health. They have tremendous impact on the health care system and may provide medical health benefits including the prevention and/or treatment of disease and various physiological disorders such as Andropause or Male menopause. Phytonutrients play a positive role by maintaining and modulating immune function to prevent specific diseases. Being natural products, they hold a great promise in clinical therapy. Phytonutrients in Nutrease powder are the plant nutrients with specific biological activities that support human health. Some of the important bioactive phytonutrients in Nutrease powder include polyphenols, terpenoids, resveratrol, flavonoids, isoflavonoids, carotenoids, limonoids, glucosinolates, phytoestrogens, phytosterols, anthocyanins, and probiotics. They play specific pharmacological effects in human health such as anti-microbial, anti-oxidants, anti-inflammatory, anti-allergic, anti-spasmodic, anti-cancer, anti-aging, hepatoprotective, hypolipidemic, neuroprotective, hypotensive, diabetes, osteoporosis, CNS stimulant, analgesic, protection from UVB-induced carcinogenesis, immuno-modulator, and carminative. This article reviews the current available scientific literature regarding the effect of Nutrease powder as an effective supplementation in Male & Female Fertility.
My talk at BASF Science Symposium: sustainable food chain - from field to table, Jun 23-24, 2015, Chicago.
Notes and acknowledgements at http://kamounlab.tumblr.com/post/122151022390/plant-pathology-in-the-post-genomics-era
Foodomics - the application of advanced omics technologies to understand the molecular and genetics level in food and correleate with nutrition and authenticationn purposes. (getnet)
Metagenomics to Unlock the Biotechnological Potential of Marine Environments by Michele de Cássia Pereira e Silva in Examines in Marine C Biology & Oceanography
Nutrients present in Nutrease powder play an important role in maintaining the normal functions of the human body. The major nutrients present in Nutrease powder include Natural carbohydrates, proteins, lipids, vitamins, and minerals. Besides these, there are some bioactive food components known as “phytonutrients” that play an important role in human health. They have tremendous impact on the health care system and may provide medical health benefits including the prevention and/or treatment of disease and various physiological disorders such as Andropause or Male menopause. Phytonutrients play a positive role by maintaining and modulating immune function to prevent specific diseases. Being natural products, they hold a great promise in clinical therapy. Phytonutrients in Nutrease powder are the plant nutrients with specific biological activities that support human health. Some of the important bioactive phytonutrients in Nutrease powder include polyphenols, terpenoids, resveratrol, flavonoids, isoflavonoids, carotenoids, limonoids, glucosinolates, phytoestrogens, phytosterols, anthocyanins, and probiotics. They play specific pharmacological effects in human health such as anti-microbial, anti-oxidants, anti-inflammatory, anti-allergic, anti-spasmodic, anti-cancer, anti-aging, hepatoprotective, hypolipidemic, neuroprotective, hypotensive, diabetes, osteoporosis, CNS stimulant, analgesic, protection from UVB-induced carcinogenesis, immuno-modulator, and carminative. This article reviews the current available scientific literature regarding the effect of Nutrease powder as an effective supplementation in Male & Female Fertility.
My talk at BASF Science Symposium: sustainable food chain - from field to table, Jun 23-24, 2015, Chicago.
Notes and acknowledgements at http://kamounlab.tumblr.com/post/122151022390/plant-pathology-in-the-post-genomics-era
E-screen assay validation: evaluation of estrogenic activity by MCF7 cell cul...Agriculture Journal IJOEAR
— Natural and synthetic estrogens have been detected in rivers, lakes and estuaries in many parts of the world. Primary sources of these compounds are domestic and industrial effluents, which are not deleted after the water treatment. Estrogen has been the endocrine disruptor most researched to be very active biologically and be the etiologic agent of diverse types of cancer and other conditions such as endometriosis, precocious puberty, feminization, masculinization, sterility. In this context, we use water of 36 natural reservoirs or dams, in a bioassay to characterize their estrogenicity in culture of MCF7 cells and obtained high concentration of estrogen in samples taken in Ibiúna and Equestrian Santo Amaro / SP. However, certain concentration in our samples for most water samples from different regions was very close to the limit of quantification by bioassay and estrogen was in fmol. It has been shown that e-screen assay with MCF7 cells is a sensitive and stable tool for quantitative analysis of estrogenicity of water and can easily be developed and implemented for routine for estrogen quantification also in animal food and man, aqueous and plastics etc. Keywords— endocrine disrupters, estrogen, breast cancer cells, (MCF7) bioassay: E-screen assay
"Keeping up with the plant destroyers." My talk at The Royal Society, 7 March...Sophien Kamoun
Tackling emerging threats to animal health, food security and ecosystem resilience, The Royal Society, Monday 7 – Tuesday 8 March 2016. https://royalsociety.org/events/2016/03/emerging-fungal-threats/
World-wide data exchange in metabolomics, Wageningen, October 2016Christoph Steinbeck
Talk given at the Netherlands Institute of Ecology in Wageningen, where I describe the development of the MetaboLights database and the value of data sharing in Metabolomics and molecular Biology in General
5 phytochemical analysis of bitter melon juice; antiproliferative and apopto...BIOLOGICAL FORUM
ABSTRACT: Osteosarcoma is one of the most common cancers among adolescents and young adults. Prognosis of osteosarcoma is particularly poor, suggesting critical needs for additional drugs to improve disease outcome. In this study, osteosarcoma cell SaOS-2 was used as an in vitro model to assess the effect of bitter melon (Momordica charantia) juice (BMJ) as an anticancer agent. Fruit juice of Momordica charantia was subjected to preliminary phytochemical analysis to identify various phytoconstituents present in them such as carbohydrates, starch, flavonoids, phenols, steroids, terpenoids, alkaloids, tanins and saponins. Cell viability (MTT) results revealed that 1 to 10% (v/v) of BMJ treatment significantly (p<0.05) reduced the proliferation of SaOS-2 cells in a dose dependent manner. Antiproliferative activity of BMJ was also coupled with morphological changes in the cells with the significant induction of apoptosis as quantified by DAPI stain. Reactive oxygen species (ROS) generation was also significantly induced by BMJ treatment. Furthermore, flow cytometric analysis revealed that BMJ induced cell cycle arrest in G2/M phase with the enhancement of apoptosis. The findings of this study suggest that BMJ modulates ROS generation and cell cycle arrest in G2/M phase of SaOS-2 cells which lead to inhibition of cell proliferation and induction of apoptosis of osteosarcoma. These findings suggest that BMJ could be a potential agent for osteosarcoma treatment and anticancer drug discovery.
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.
E-screen assay validation: evaluation of estrogenic activity by MCF7 cell cul...Agriculture Journal IJOEAR
— Natural and synthetic estrogens have been detected in rivers, lakes and estuaries in many parts of the world. Primary sources of these compounds are domestic and industrial effluents, which are not deleted after the water treatment. Estrogen has been the endocrine disruptor most researched to be very active biologically and be the etiologic agent of diverse types of cancer and other conditions such as endometriosis, precocious puberty, feminization, masculinization, sterility. In this context, we use water of 36 natural reservoirs or dams, in a bioassay to characterize their estrogenicity in culture of MCF7 cells and obtained high concentration of estrogen in samples taken in Ibiúna and Equestrian Santo Amaro / SP. However, certain concentration in our samples for most water samples from different regions was very close to the limit of quantification by bioassay and estrogen was in fmol. It has been shown that e-screen assay with MCF7 cells is a sensitive and stable tool for quantitative analysis of estrogenicity of water and can easily be developed and implemented for routine for estrogen quantification also in animal food and man, aqueous and plastics etc. Keywords— endocrine disrupters, estrogen, breast cancer cells, (MCF7) bioassay: E-screen assay
"Keeping up with the plant destroyers." My talk at The Royal Society, 7 March...Sophien Kamoun
Tackling emerging threats to animal health, food security and ecosystem resilience, The Royal Society, Monday 7 – Tuesday 8 March 2016. https://royalsociety.org/events/2016/03/emerging-fungal-threats/
World-wide data exchange in metabolomics, Wageningen, October 2016Christoph Steinbeck
Talk given at the Netherlands Institute of Ecology in Wageningen, where I describe the development of the MetaboLights database and the value of data sharing in Metabolomics and molecular Biology in General
5 phytochemical analysis of bitter melon juice; antiproliferative and apopto...BIOLOGICAL FORUM
ABSTRACT: Osteosarcoma is one of the most common cancers among adolescents and young adults. Prognosis of osteosarcoma is particularly poor, suggesting critical needs for additional drugs to improve disease outcome. In this study, osteosarcoma cell SaOS-2 was used as an in vitro model to assess the effect of bitter melon (Momordica charantia) juice (BMJ) as an anticancer agent. Fruit juice of Momordica charantia was subjected to preliminary phytochemical analysis to identify various phytoconstituents present in them such as carbohydrates, starch, flavonoids, phenols, steroids, terpenoids, alkaloids, tanins and saponins. Cell viability (MTT) results revealed that 1 to 10% (v/v) of BMJ treatment significantly (p<0.05) reduced the proliferation of SaOS-2 cells in a dose dependent manner. Antiproliferative activity of BMJ was also coupled with morphological changes in the cells with the significant induction of apoptosis as quantified by DAPI stain. Reactive oxygen species (ROS) generation was also significantly induced by BMJ treatment. Furthermore, flow cytometric analysis revealed that BMJ induced cell cycle arrest in G2/M phase with the enhancement of apoptosis. The findings of this study suggest that BMJ modulates ROS generation and cell cycle arrest in G2/M phase of SaOS-2 cells which lead to inhibition of cell proliferation and induction of apoptosis of osteosarcoma. These findings suggest that BMJ could be a potential agent for osteosarcoma treatment and anticancer drug discovery.
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.
Cytogenetic, Hematological and Enzymes Levels Parameters in the Biomonitoring...inventionjournals
Studies have demonstrated genotoxic effects by the presence of micronucleus in exfoliated cells from the buccal mucosa of agricultural workers exposed to pesticides. This study has assessed the genotoxic effects of pesticides on 61 agricultural workers from the state of Piauí, Brazil. 31 individuals were exposed to pesticides and 30 are from the same area, but were not involved in pesticides application. Cytogenetic damage were evaluated through micronucleus test in cells from the buccal mucosa and some parameters such as hematological and levels of enzymes. Exposed individuals exhibited cytogenetic damage with increased number of micronuclei in cells from the buccal mucosa in comparison with subjects from the control group with significant statistical difference (P < 0.01). We perceive that there is a statistically no significant (P > 0.05) increase in levels of plasmatic and eritrocytaireacetylcholinesterase and no statistically significant increase of phosphatase alkaline were detected in exposed workers in relation to the control group. No association was found in relation to smoking habits, alcohol consumption, protection utensils and the biomarkers analyzed or the biochemical analysis. Analysis of variance revealed a correlation between occupational exposure to pesticides of workers in Piauí and the presence of micronuclei (P < 0.05).
Cytogenetic, Hematological and Enzymes Levels Parameters in the Biomonitoring...inventionjournals
Studies have demonstrated genotoxic effects by the presence of micronucleus in exfoliated cells from the buccal mucosa of agricultural workers exposed to pesticides. This study has assessed the genotoxic effects of pesticides on 61 agricultural workers from the state of Piauí, Brazil. 31 individuals were exposed to pesticides and 30 are from the same area, but were not involved in pesticides application. Cytogenetic damage were evaluated through micronucleus test in cells from the buccal mucosa and some parameters such as hematological and levels of enzymes. Exposed individuals exhibited cytogenetic damage with increased number of micronuclei in cells from the buccal mucosa in comparison with subjects from the control group with significant statistical difference (P < 0.01). We perceive that there is a statistically no significant (P > 0.05) increase in levels of plasmatic and eritrocytaireacetylcholinesterase and no statistically significant increase of phosphatase alkaline were detected in exposed workers in relation to the control group. No association was found in relation to smoking habits, alcohol consumption, protection utensils and the biomarkers analyzed or the biochemical analysis. Analysis of variance revealed a correlation between occupational exposure to pesticides of workers in Piauí and the presence of micronuclei (P < 0.05).
Cytogenetic, Hematological and Enzymes Levels Parameters in the Biomonitoring...inventionjournals
Studies have demonstrated genotoxic effects by the presence of micronucleus in exfoliated cells from the buccal mucosa of agricultural workers exposed to pesticides. This study has assessed the genotoxic effects of pesticides on 61 agricultural workers from the state of Piauí, Brazil. 31 individuals were exposed to pesticides and 30 are from the same area, but were not involved in pesticides application. Cytogenetic damage were evaluated through micronucleus test in cells from the buccal mucosa and some parameters such as hematological and levels of enzymes. Exposed individuals exhibited cytogenetic damage with increased number of micronuclei in cells from the buccal mucosa in comparison with subjects from the control group with significant statistical difference (P < 0.01). We perceive that there is a statistically no significant (P > 0.05) increase in levels of plasmatic and eritrocytaireacetylcholinesterase and no statistically significant increase of phosphatase alkaline were detected in exposed workers in relation to the control group. No association was found in relation to smoking habits, alcohol consumption, protection utensils and the biomarkers analyzed or the biochemical analysis. Analysis of variance revealed a correlation between occupational exposure to pesticides of workers in Piauí and the presence of micronuclei (P < 0.05).
Applications of bioinformatics, main by kk sahuKAUSHAL SAHU
Introduction
Goals of Bioinformatics
Bioinformatics & Human Genome
Project
What can we do using bioinformatics ?
Applications of bioinformatics in various fields
1) Medicine
2) Evolutionary studies
3) Agriculture
4) Microbiology
5) Biotechnology
Conclusion
References
31.Medicinal uses of poisonous fishes..pdfsalehaasawer
Science makes new discoveries everyday that advance human health and well being. According to new researches fish poison have huge medical importance. These poisons are examined in several animal models.
Biological diversity, or biodiversity, is the scientific term for the variety and variability of life on Earth. Biodiversity is the key indicator of the health of an ecosystem. Every living thing, including man, is involved in these complex networks of interdependent relationships, which are called ecosystems.
Like all healthy ecosystems, Richness of microbiota species characterizes the GI microbiome in healthy individuals. Conversely, a loss in species diversity is a common finding in several disease states. Microbiota Biodiversity helps us : 1- Combat aggressions from other microorganisms, 2- Maintaining the wholeness of the intestinal mucosa. 3- Plays an important role in the immune system, 4- Performing a barrier effect.5- A healthy and balanced gut microbiota is key to ensuring proper digestive functioning. A gut out of balance means a body out of balance which means illness including Inflammation, Allergies, Infections, Nutrient deficiencies, Weight Gain, Asthma-allergies – Autoimmunity
• Arthritis, Metabolic Bone disease, Skin problems e.g. eczema, rosacia, Mood disorders - Cognitive decline-Alzheimers and Cancer.
Metabolomics: The Next Generation of Biochemistry Metabolon, Inc.
This brief eBook explores the benefits of incorporating the science of metabolomics into contemporary biology research as a stand-alone tool or as a compliment to genomics or other types of molecular biology research.
GC/MS analysis and In-vitro Antioxidant activity of methanol extract of Uloth...IOSRJPBS
The determination of phytochemical constituents, total phenol, flavonoid contents and antioxidant assays of methanol extract of Ulothrix flacca and its main constituent dimethyl sulfone was studied. The mass spectra of the compounds were matched with the NIST library. The GC-MS analysis of methanol extracts of Ulothrix flacca showed sixteen peaks. Of all the sixteen chemical compounds revealed from the GC-MS analysis of Ulothrix flacca, Dimethyl Sulfone (C2H6O2S) (RT-8.9), 4-Bromobenzoic Acid, 2-Chlorophenyl Ester (C13H8BrClO2) (RT-12.642), Tetradecanoic Acid, 10,13-Dimethyl-, Methyl Ester (C17H34O2) (RT-18.669) are the three major components. The methanol extracts of Ulothrix flacca possess phenolic and flavonoid content of (5.74 ± 0.45mg Gallic acid equivalent (GAE)/g Wt, and 12.58 ± 1.52mg quercetin eq/g wt) respectively. Antioxidant activity was determined using 1,1-diphenyl-2-picryl hydrazyl (DPPH) free radical, for evaluating free radicle scavenging activity, ABTS radical cation scavenging activity, Ferric reducing antioxidant power (FRAP) assay, Phosphomolybdenum assay and Metal chelating activity using BHT, Rutin and Quercetin. The highest radicle scavenging activity was shown by dimethyl sulfone (15.156mg/ml), which is higher than the BHT and Rutin. In vitro antioxidant activity of methanolic extract of Ulothrix flacca and Dimethyl sulfone showed an increase with increasing concentration indicating positive association with the total phenolic and flavonoid contents of the extract, which could be considered for future applications in medicine, dietary supplements ,cosmetics or food industries.
Labeorohita, is a fish of the carp family Cyprinidea, found commonly in rivers and freshwater lakes in and around South Asia and South-East Asia. IT is used as a major diet all over India. Hence its culture is done at a heavy ratio accordance to other fishes. Translationally Controlled Tumor Protein found in Labeorohitahas a very similar polypeptide chain accordance with human. Human TCTP has a great effect on growth. Due to the unavailability of the Tertiary structure of TCTP of Labeorohita, clear functional activity is unable to detect. The following work is a try to build a tertiary structure and detecting its active sites by docking it with different active proteins.
DOI:10.21276/ijlssr.2016.2.4.20
ABSTRACT- The present investigation was carried out to the effect of dimethoate on histopathological changes in
kidney of freshwater fish, Garra mullya. Fishes was exposed to sub lethal concentration of dimethoate (0.0238ppm of
96hrs.) for 7, 14, 21 days. Fishes exposed to dimethoate were characterized by loosening of haemopoietic tissue,
uriniferous tubules have lost their original appearance, vacuolated cytoplasm, degeneration in the epithelial cells of renal
tubule, narrowing of the tubular lumen and damaged glomeruli. The lesions in the vital organ might have resulted in
physiological and metabolical dysregulations. In chronic treatment of dimethoate exposure may pose serious threat to fish
health and affect their population. Key-words- Dimethoate, Histopathology, Kidney, Garra mullya
Many Gram negative bacteria, especially Aeromonas hydrophila are notorious for their heightened capacity to acquire and exchange antibiotic resistance genes and consequently, are commonly targeted as indicator organism for monitoring antimicrobial resistance in aquatic milieus. This study was aimed to investigate the prevalence and drug resistance patterns of Aeromonas hydrophila isolated from farm raised catfish at Epe fish farm, Lagos State, Nigeria. Swabs were aseptically taken from the kidney, intestine, liver, gills, and skin after dissecting the fish samples. The specimens were bacteriologically analyzed. After series of biochemical test, the isolated bacteria were identified presumptively according to Bergey’s Manual of determinative bacteriology, 7th Edition. Fifty-seven (57) Aeromonas hydrophila were recovered out of seventy-one (71) bacterial isolated from the 35 fish samples collected from the fish farms studied. The study reveals multiple antibiotics resistance pattern among the isolates as Aeromonas hydrophila were 100% resistant to Streptomycin, Oxytetracycline, Chloranphenicol, Pefloxacin, Ofloxacin and 70, 65 and 55% resistant to Gentamycin, Amoxycillin and Erythromycin respectively. However, all the isolates were 100% susceptible to Ciprofloxacin and Cotrimoxazole only. The presence of multidrug-resistant Aeromonas hydrophila in fish could be a vehicle of horizontal gene transfer to previously susceptible bacteria and these could constitute a serious public health hazard to human and animal in the environment. Thus, the use of antibiotics in catfish production as growth promoter or disease prevention should be discouraged and some safer, biological alternatives strategies should suffice to mitigate bacteria drug resistance and its associated problems.
Similar to International Journal of Proteomics & Bioinformatics (20)
A 5-year old boy, with an established diagnosis of a topic
dermatitis, previously treated by topical corticosteroids and emollient cream with a good improvement, developed widespread papules on his legs, hands and forearm that appeared 5 months ago.
Methods: Retrospectively, the file records of the patients who underwent sleeve gastrectomy were examined. Demographic features, Body Mass Index (BMI), the mouth opening, Mallampati score, thyromental distance, sternomental distance, neck circumference measurements and videolaryngoscopic examination results were recorded Results: In a total of 140 consecutive patients (58 male, 82 female) were included in the study. The mean age of the study participants was 35.40 ± 9.78 and the mean BMI of the patients was 44.33 ± 7.52 kg/m2
. The mean mouth opening of the patients was 4.82 ± 0.54 cm
and the mean neck circumference was 43.52 ± 4.66 cm. The mean thyromental distance was 8.02 ± 1.00 cm and the mean sternomental distance was16.58 ± 1.53 cm. Difficult intubation was determined in 8 (5.7%) patients. In logistic regression analysis, age (p : 0.446), gender (p : 0.371), BMI (p : 0.947), snoring (p : 0.567), sleep apnea (p : 0.218), mouth opening (p : 0.687), thyromental distance (p :0.557), sternomental (p : 0.596) and neck circumference (p : 0.838) were not the independent predictors of difficult intubation. However, Mallampati score (p : 0.001) and preoperative direct laryngoscopy findings (p : 0.037) performed in outpatient clinic were the significant
predictors of difficult intubation. Interestingly, all patients with grade 4 laryngoscopy findings had difficult intubation.
Introduction: Laparoscopic surgery has been performed in Mexico since 1989, but no reports about training tendencies exist. We conducted a national survey in 2015, and here we report the results concerning training characteristics during the surgical residence of the respondents. Materials and Methods: A prospective study was conducted through a survey questioning demographic data, laparoscopic training during pre and post surgical residency and other of areas of laparoscopic practice. The sample was calculated and survey piloted before
application. Special interest in this report was placed on type and quality of training received. Data are reported in percentages.
Heterotopic Ossification (HO) is defined as pathological bone formation at locations where bone normally does not exist. The
presence of HO has been found to be a rare complication after stroke in several studies, whereas there are only sporadic references relating HO to Cerebral Palsy (CP) and few for CP and stroke. No effective treatment for HO has yet been found, whereas the cellular and molecular mechanisms have not been completely understood. Therefore, increased awareness among physicians is required, as a challenge for early diagnosis and treatment. A case of a male patient with CP, who developed HO on the paretichip joint following an ischemic stroke is presented.
Objectives: To assess the practice of food hygiene and safety, and its associated factors among street food vendors in urban areas of Shashemane, West Arsi Zone, Oromia Ethiopia, 2019.
Methods: Cross-sectional study design was applied from December 28, 2019 to January 27, 2020. Data was collected from 120 food handlers, which were selected by purposive sampling techniques. Information was gathered from interview and field observation by conducting food safety survey and using questionnaires via face to face interview. The collected data was entered using Epi Data 3.1 and finally, it was analyzed using SPSS VERSION 20.
A Division I football athlete experienced acute posterior leg pain while pushing off on the line of scrimmage. Ultrasound (US) showed a midsubstance plantaris tendon rupture, an injury that, to our knowledge, has only been described once before in the medical literature [1]. US was also used to assist with rehab progression and return to previous level of activity, which was achieved three weeks after the injury. While there currently are no guidelines regarding return to sport after this injury, this case demonstrates that once pain is controlled and ROM restored, progression through rehabilitation and return to elite level sport is simply based on symptoms.
Type 1 Diabetes (T1D), is a severe disease, representing 5-10% of all reported cases of diabetes worldwide. Fulminant Type 1 Diabetes Mellitus (FT1D) is a subtype of type 1 diabetes mellitus that is largely characterized by the abrupt onset of Diabetic Ketoacidosis (DKA) and severe hyperglycemia without insulin defi ciency. Viral infections have been hypothesized to play a major role in the pathogenesis of Fulminant Type 1 Diabetes Mellitus (FT1D) through the complete and rapid destruction of pancreatic beta cells. Coxsackie viral infection has been detected in islets of 50% of the pancreatic tissue recovered from recent-onset Type 1 Diabetes (T1D) patients. In this report we have highlighted a case where the patient developed a Group B Coxsackie virus infection culminating in the development of Fulminant Type 1 Diabetes Mellitus (FT1D).
Methods: Cercariae are released by infected water snails. To determine the occurrence of cercariae-emitting snails in SchleswigHolstein, 155 public bathing places were visited and searched for fresh water snails. Family and genus of the collected snails were determined and the snails were examined for the shedding of cercariae, using a standard method and a newly developed method.
Objective: To generate preliminary information about of enteroviruses and Enterovirus 71 (EV71) in patients with aseptic meningitis in Khartoum State, Sudan.
Method: Cerebrospinal fluid specimens were collected from 89 aseptic meningitis patients from different Khartoum Hospitals
(Mohammed Alamin Hamid Hospital, Soba Teaching Hospital, Omdurman Military Hospital, Alban Gadeed Teaching Hospital and Police Hospital) within February to May 2015. Among these 89 patients, 43 (48%) were males and 46 (52%) were females. The patient’s age ranged between 1 day and 30 years old. The collected specimens were assayed to detect enteroviruses and EV71 RNA using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) technique
Femoral hernias, comprise 2% to 4% of all hernias in the inguinal region, and occur most commonly in women. Th ey present typically with a mass below the level of the inguinal ligament. The sac may contain preperitoneal fat, omentum, small bowel, or other structures and have a high rate of incarceration and strangulation due to the small size of the hernia neck orifice, requiring emergency surgery. We present the case of a 54-year-old female patient with intestinal occlusion due to incarcerated femoral hernia, repaired by laparoscopic approach, that gave the patient the opportunity to attend her daughter’s wedding the same day.
Small Supernumerary Marker Chromosome (sSMC) is a rare genetic condition marked by the presence of an extra chromosome to the 46 human chromosomes. This case report describes a 4 year old child with SSMC on the 46th chromosome. The child presented with delayed speech and language development, seizures and mild developmental delay. Speech and Language evaluation was carried out and management options are discussed.
A catheter is a thin tube made from medical grade materials that serve a broad range of functions, but mainly catheters are medical devices that can be inserted in the body to treat disease or perform surgical procedures. Catheters have been inserted into body cavities, ducts, or vessels to allow for drainage, administration of therapeutic fluids or gases, operational access for surgery. Catheters help perform tasks in various systems such as cardiovascular, urological, gastrointestinal, neurovascular, and ophthalmic systems. A dataset of 12 patients with varying “weights” and “heights” was recorded along with the lengths of their catheter tubes. This data set was found from two revered statistical textbooks on linear regression and the Department of Scientific Computing at Florida State University. This data set was not able to be linked to any particular clinical or experimental research studies, but the data set can be used to help catheter manufacturers and medical professionals better decide on what particular catheter lengths to use for patients knowing only their height & weight. These research insights could be helpful to healthcare professionals that have patients with incomplete or no healthcare records
to decide what catheter length to use. The main investigative inquiry that needed to be answered was how does patient weight & height influence catheter length together and separately? We conducted linear regression and other statistical analysis procedures in R program & Microsoft Excel and discovered that this data exhibited a quality called multi collinearity. With multi collinearity, all predictors (2 or more
independent variables) are not significant in an all encompassing linear aggression, but the predictors might be significant in their own individual linear regressions. Individual linear regression analyses were conducted for both patient height & weight to see how much they both contribute to varying catheter length. Patient weight was found to be more impatful than patient height in relationship to catheter length, even though height and weight are a classical example of multi collinearity predictors.
Bovine mastitis has a negative impact through economic losses in the dairy sector across the globe. A cross sectional study was carried out from September 2015 to July 2016 to determine the prevalence of bovine mastitis, associated risk factors and isolation of major causative bacteria in lactating dairy cows in selected districts of central highland of Ethiopia. A total of 304 lactating cows selected randomly from five districts were screened by California Mastitis Test (CMT) for subclinical mastitis. Based on CMT result and clinical examination, over all prevalence of mastitis at cow level was 70.62% (214/304).
Two hundred fourteen milk samples collected from CMT positive cows were cultured for isolation of major causative bacteria. From 214 milk samples,187 were culture positive and the most prevalent isolates were Staphylococcus aureus 42.25% (79/187) followed by Streptococcus agalactiae 14.43%
(27/187). Other bacterial isolates were included Coagulase Negative Staphylococcus species 12.83% (24/187), Streptococcus dysgalactiae 5.88% (11/187), Escherichia coli 13.38% (25/187) and Entrococcus feacalis 11.23% (21/187) were also isolated. Moreover, age, parity number, visible teat abnormalities,husbandry practice, barn fl oor status and milking hygiene were considered as risk factors for the occurrence of bovine mastitis and they were found significantly associated with the occurrence of mastitis (p < 0.05). The findings of this study warrants the need for strategic approach including dairy extension that focus on enhancing dairy farmers’ awareness and practice of hygienic milking, regular screening for subclinical mastitis, dry cow therapy and culling of chronically infected cows.
Kratom is an herbal product that is derived from Southeast Asian Mitragyna speciose tree leaves [1-10]. This compound is used for many purposes such as stimulation, euphoria, or analgesia [1-10]. It has been recently identified as a drug of abuse by the United States Drug Enforcement Administration [2,8]. Side-effects from this compound have not been well documented. We describe a case of a 36-year-old female who develop nephrotoxicity after taking an herbal supplement. She took kratom as an adjunctive therapy for back pain management. She developed right upper quadrant pain and nausea. Laboratory tests showed elevated liver enzymes without evidence of bile duct obstruction. Liver enzymes normalized several weeks after Kratom discontinuation. We advise clinicians to be vigilant about Kratom’s hepatotoxic potential on patient health.
The assessment, diagnosis and treatment of critically ill patients is extremely challenging. Patients often deteriorate whilst being
reviewed and their rapidly changing pathophysiology barrages healthcare professionals with new data. Furthermore, comprehensive assessments must be postponed until the patient has been stabilised. So, important data and interventions are often missed in the heat of the moment. In emergency situations, suboptimal management decisions may cause signifi cant morbidity and mortality. Fortunately, standardisation and careful design of documentation (i.e. proformas and checklists) can enhance patient safety. So, I have developed a series of checklist proformas to guide the assessment of critically ill patients. These proformas also promote the systematic recording and presentation of information to facilitate the retrieval of the precise data required for the management for critically ill patients. The proformas have been modifi ed extensively over the last twenty years based on my personal experience and extensive consultation with colleagues in several world-renowned centres of excellence. The proformas were originally developed for use in the intensive therapy unit
or high dependency unit. However, they have been adapted for use by outreach teams reviewing patients admitted outside of critical care areas. The use of these tools can direct eff orts to provide appropriate organ support and provides a framework for diagnostic reasoning.
Systemic Hypertension (HTN) accounts for the largest amount of attributable Cardiovascular (CV) mortality worldwide. There are several factors responsible for the development of HTN and its CV complications. Multicenter trials revealed that risk factors responsible for Micro Vascular Disease (MVD) are similar for those attributable to Coronary Artery Disease (CAD) which include tobacco use, unhealthy cholesterol levels, HTN, obesity and overweight, physical inactivity, unhealthy diet, diabetes, insulin resistance, increasing age and genetic predisposition. In addition, the defective release of Nitric Oxide (NO) could be a putative candidate for HTN and MVD. This study reviewed the risk stratification of hypertensive population employing cardiac imaging modalities which are of crucial importance
in diagnosis. It further emphasized the proper used of cardiac imaging to determine patients at increased CV risk and identify the management strategy. It is now known that NO has an important effect on blood pressure, and the basal release of endothelial Nitric Oxide (eNOS) in HTN may be reduced. Although there are different forms of eNOS gene allele, there is no solid data revealing the potential role of the polymorphism of the eNOS in patients with HTN and coronary vascular diseases. In the present article, the prevalence of eNOS G298 allele in hypertensive patients with micro vascular angina will be demonstrated. This review provides an update on appropriate and justified use of non-invasive imaging tests in hypertensive patients and its important role in proper diagnosis of MVD and CAD. Second, eNOS gene allele and its relation to essential hypertension and angina pectoris are also highlighted.
Methods: Two groups were selected by non-probability random sampling technique including case group of 154 patients with
suspected dengue (fever>2days and <10days) and control group of 146 patients with febrile illness other than dengue. Clinical,hematological and serologic markers of cases and control groups were analyzed. The frequency distribution was used to compare categorical serologic markers and paired sample T test was applied for hematologic variables before and after treatment of dengue using SPSS version 21.
Researchers from Utrecht recently published yet another paper on the use of Magnetic Resonance Imaging (MRI)demonstrating an additional failed attempt to understand the importance of qualitative versus quantitative imaging, and anatomic versus physiologic imaging. Th e implications of this failure here cannot be overstated.
Introduction: Stroke is an even more dramatic major public health problem in young people. Goal of the study: Contribute to the knowledge of strokes in young people. Methodology: This was a retrospective study carried out over a period of 02 years (January 2017 to December 2018) including the files of patients aged 18 to 49 years hospitalized for any suspected case of stroke in the Neurology department of the University Hospital
Center of the Sino-Central African Friendship (CHUSCA) of Bangui.
Background: This report describes a unique case of a patient that developed psychotic symptoms believed to be secondary
to a tentorial meningioma with associated hydrocephalus. These psychotic symptoms subsequently abated with placement of a
ventriculoperitoneal shunt. Case description: 60-year-old female was admitted to an inpatient psychiatric facility on a psychiatric involuntary commitment petition due to progressive paranoia, homicidal ideation and psychosis. The work up showed a calcified six cm tentorial meningioma with associated hydrocephalus. The patient initially rejected treatment but later became amenable to placement of Ventriculoperitoneal Shunt
(VPS).
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Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
Anti ulcer drugs and their Advance pharmacology ||
Anti-ulcer drugs are medications used to prevent and treat ulcers in the stomach and upper part of the small intestine (duodenal ulcers). These ulcers are often caused by an imbalance between stomach acid and the mucosal lining, which protects the stomach lining.
||Scope: Overview of various classes of anti-ulcer drugs, their mechanisms of action, indications, side effects, and clinical considerations.
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdfJim Jacob Roy
Cardiac conduction defects can occur due to various causes.
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Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
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2. International Journal of Proteomics & Bioinformatics
SCIRES Literature - Volume 4 Issue 1 - www.scireslit.com Page - 016
INTRODUCTION
Aquatic ecosystems have been faced with several threats from
complex mixture of contaminants from industries, anthropogenic
perturbations and other stressors in recent years. And for several
decades, the presence of these environmental contaminants in
aquatic biotic and abiotic samples has been measured using chemical
analytical techniques.
Environmental stress, as well as a variety of physical conditions,
may sometimes induce the synthesis of certain proteins in fish. Some
of these proteins are believed to play a role in protecting the cell from
the damage, which may result from environmental perturbations,
while others are involved in the regulation of various genes. The
best-known representatives of this group are the stress proteins (also
known as Heat Shock Proteins, (HSPs) and Metallothioneins (MTs).
Proteomics has also been applied to specific aspects of seafood
research and technology. Proteomics applications related with
seafood production and safety has been classified in three main items:
(i) food safety studies, (ii) authentication and taxonomic applications,
and (iii) nutritional aspects [1]. Proteins can be related to pollution
initiation, safety, quality and nutrition topics, constituting in this
sense important biomarkers for the detection of pollution, food safety,
and nutritional value [2], as well as monitoring nutritional differences
between wild and farmed species as applied in nutriproteomics [3].
Muscle plays a central role in whole-body protein metabolism
by serving as the principal reservoir for amino acids to maintain
protein synthesis in vital tissues and organs [4]. Skeletal muscle
fibers represent one of the most abundant cell types in vertebrates [5]
and contractile fibers of skeletal muscle tissues provide coordinated
excitation-contraction-relaxation cycles for voluntary movements
and postural control [6]. Besides playing a central physiological role
in heat homeostasis, it usually presenting itself as a crucial metabolic
tissue that integrates various biochemical pathways [7]. Therefore,
muscle proteomics aims at global identification, cataloging and
biochemical characterization of the entire protein complement of the
voluntary contractile tissue as well as have the potential to identify
novel proteins which could serve as biomarkers for many aspects
including fish physiology and growth, flesh quality, food safety and
aquatic environmental monitoring [8,9].
The most common methods for protein identification are Matrix-
Assisted Laser Desorption Ionization- Time of Flight (MALDI-
TOF) coupled with 2 Dimensional gel Electrophoresis (2DE) and
Electrospray Ionization Tandem Mass Spectrometry (ESI-MS/MS).
MALDI-TOF provides high sequenced coverage in proteins from
sequenced genomes, while ESI-MS/MS is the method of choice in
the case of genomes with no fully sequencing information available.
The growth of mass spectrometry facilities, powerful bioinformatics
tools, and availability of genomic information in research centres
and industries can make possible that high throughput proteomics
will feature in aquaculture and fisheries in the near future. The high-
throughput proteomic approaches such as isobaric Tagging for
Relative and Absolute Quantitation (iTRAQ), has been successfully
used to study the responses by fish to aquatic pollutants, including
androgen receptor agonists/antagonists [10]. This high-through
proteomics has been successfully applied to identify altered protein
expression in the muscle of rainbow trout Oncorhynchus mykiss
during spawning and was associated with biochemical processes
involved in muscle deterioration.
PROTEOMIC IN FISH
It is increasingly important to profile proteins in order to
understand biological processes in a post genomic era as the dynamics
of proteins between cells at different times and under different
environmental conditions provide an actual biological phenotype.
Proteomics studies are built upon the foundations of genomics, in
such way that a lack of genomic information on a particular species
can substantially limit the success in protein identification [11]. As
such, functional genomics and proteomic technologies have allowed
simultaneous biological study of thousands of genes or proteins. In
particular, the presence of post translational modifications in proteins
further highlights the importance of proteomic analysis which is not
replaceable by other genomic approaches [12].
TECHNIQUES AND APPROACHES IN IDEN-
TIFICATION OF POTENTIAL PROTEIN
MARKERS
Different molecular approaches [biochemical assays, Enzyme
Linked Immuno-Sorbent Assays (ELISA), spectrophotometric,
fluorometric measurement, differential pulsed polarography, liquid
chromatography, atomic absorption spectrometry]. However,
methodological approaches able to detect subtle changes in
the expression of individual proteins and amino acid sequence
modifications, constitute an excellent tool [13,14], which can be
explanatory of a biological situation. Recent report shows exponential
developments concerning proteome analytical techniques and
sample preparation [15]. Proteomics is essentially based on classical
“Analytical Chemistry” strategies: separation by electrophoresis
ABSTRACT
Studying the expression of proteins in Chrysichthys nigrodigitatus muscle, is essential to understand the biological, physiological and
ecological aspects that may be of advantage in ecotoxicology as a tool for biomonitoring the effects of environmental pollution, as well as
food safety. This study was aimed to accomplish a systematic characterization of the muscle proteome as well as to identify a putative
set of protein biomarkers in C. nigrodigitatus to environmental pollution in Ologe and Badagry lagoons. Fifteen fish samples were used
as representative of the population for proteomics analysis. 116 proteins was expressed with 70 up-regulated, 25 down-regulated from
Ologe Lagoon and 30 up-regulated, 17 down-regulated proteins from Badagry Lagoon expressed in the muscle of C. nigrodigitatus in
response to environmental stressors using iTRAQ, while 8 protein spots from 2-D gels, representing 8 proteins with 2 up-regulated and 6
down-regulated, have been identified using MALDI TOF/TOF MS. Pearson correlation revealed significant correlation (p < 0.05) between
environmental variables and protein markers. The investigation revealed that the expressed proteins in the muscle of C. nigrodigitatus
served as a prognostic tool to assess the fish health and pollution status, which was observed that the fish were physiologically perturbed
by environmental stressors in Ologe and Badagry lagoons.
Keywords: Chrysichthys nigrodigitatus; Proteomics; 2-D electrophoresis; MALDI-TOF-MS, iTRAQ, Lagoons
3. International Journal of Proteomics & Bioinformatics
SCIRES Literature - Volume 4 Issue 1 - www.scireslit.com Page - 017
techniques, identification and quantitative analysis through mass
spectrometry.
PROTEOMICS AS ENVIRONMENTAL MARK-
ER OF POLLUTION
Proteomics applied to assess the biological effects of pollutants in
marine organisms is beginning to reveal some of the systemic changes
that occur on the cellular level. First, even a qualitative description
of cellular changes through PES shows that the systemic changes
occurring during exposure are pollutant-specific. Second, proteins
common to many pollutant-stress responses include oxidative stress
proteins, cytoskeletal proteins, chaperones, proteases, and proteins
involved in the detoxification of xenobiotics as well as β-oxidation.
Together, these changes suggest that the production of ROS leads to
the denaturation of proteins as well as wide-ranging modifications of
cytoskeletal elements. Finally, PTMs present a novel frontier to assess
the biological effects of pollutants. Among the PTMs that are likely to
be the most important are those that increase with oxidative stress,
e.g. Carbonylation and glutathionylation, and that are indicators of
protein denaturation, e.g. ubiquitination.
Fisheries have received a great deal of concern as a potential
source of heavy metal contamination to humans because of its wide
consumption of fisheries [16,17]. In West Africa, the Bagrid catfish
(Chrysichthys nigrodigitatus) are commercially important fish species
that are widely consumed and continue to attract high patronage.
Chrysichthys nigrodigitatus on the other hand, is a demersal omnivore
inhabiting the bottom of shallow waters, rivers and lagoons [18].
The Ologe and Badagry Lagoons, a home for a wide variety
of fishery resources have been found to be subjected to many
anthropogenic sources of contamination by virtue of their socio-
economic importance and proximity to urban centres, heavy-
industrial and use for several agricultural activities. Previous studies
[19-23] revealed varying levels of contaminants in these ecosystems
including heavy metals, pesticides and polycyclic aromatic
hydrocarbons. These compounds can be absorbed and accumulated
in the edible parts of lagoon biota, thus entering the human food
chain and posing a public health problem. However, there is paucity
of information on the proteome compositions of most edible aquatic
biota from Ologe and Badagry Lagoons. Therefore, proteomic studies
would be quite useful as additional information during environmental
contamination studies in these ecosystems.
The aim of this study was to evaluate the potential of proteomics
as a tool for biomarker discovery, providing characterization
of the proteome sensitive pollution markers in the muscle of C.
nigrodigitatus captured from the wild. Thus, the application of
proteomic hypothesis-independent approach was to discover the
protein expression patterns underlying the effects of environmental
stressors in form of admixtures of chemical pollutants from several
anthropogenic activities in an attempt of identifying a putative set of
protein biomarkers.
MATERIALS AND METHODS
Ologe and Badagry Lagoons are situated within the Lagos lagoon
complex in Lagos State of Southwestern region of Nigeria (Figure
1). Ologe Lagoon is a freshwater body which opens into the Atlantic
Ocean via the Badagry creeks and the Lagos harbour. These Lagoons
meet several socio-economic needs such as aquaculture, fishing, sand
dredging and drainage across various towns and villages bordering
it [19]. By virtue of their position, (with several industries and other
anthropogenic activities around it), have been impacted by partially
treated and untreated industrial effluents.
Experimental set-up
Fish (Chrysichthys nigrodigitatus) samples were collected using a
trap basket slightly above the bottom of the lagoon, where the water
was shallow at depth of 3.5 m from the eight (8) established stations
in Ologe and Badagry Lagoons between October 2013- April 2015.
Water quality and heavy metal analyses
Water quality parameters were characterized following APHA
standard protocols [24]. Two (2) g of the dried sediment was taken
in a digestion beaker and 10 mL of mixture of HCl and HNO3
in
the ratio 3:1 was added and then digested at 95°C for 2 h. The metal
concentrations of both the digested water and sediment samples were
quantified using GBC (Savant AA Sigma) flame Atomic Absorption
Spectrometer (AAS). All chemical regents used herein were of
analytical reagent grade (Merck, United State). The glassware was
pre-cleaned with nitric acid and rinsed with double distilled water.
Preparation of protein extracts and quantification
One hundred (100) mg of muscle tissue were collected from
Chrysichthys nigrodigitatus and minced using a clean scalpel. The
tissue was homogenized with 1 mL of PBS and spinned at 3,000 rpm
for 10 min at 40
C. The supernatants were removed, and then 1 ml
of whole cell lysis buffer was added to the 100 mg of homogenized
tissue. The Whole Cell Lysis (WCL) is composed of 50 mM NaCl,
1M HEPES (pH 7.8), 5 mg/ml leupeptin, 10 mg/ml Pepstatin, 250
mM DDT, Tris, 100 mM PMSF, 40 mM Na3
VO4
and distilled water.
Sonication for 4x8 at output level 5, Spinned at 13,000 rpm for 10
min at 40
C, the supernatant was removed, saved in another tube
and centrifuged. The mixture was incubated in ice for 30 additional
minutes, and then divided in 100 μL aliquots that were stored at -80°C
until needed. The quantification results and the quality of the protein
extracts was performed by the modified Bradford method, measuring
the samples extracts in duplicates and using Bovine Serum Albumin
(BSA) as protein standard. The results of the quantification were
checked by running 25 μg of each protein extract on 12.5% vertical
SDS-PAGE gels according to Laemmli’s protocol [25].
For 2D-PAGE, protein extracts were diluted in rehydration buffer
Figure 1: Sampling stations at Ologe and Badagry Lagoons
4. International Journal of Proteomics & Bioinformatics
SCIRES Literature - Volume 4 Issue 1 - www.scireslit.com Page - 018
with composition 60 mM urea, 2 M thiourea, 2% CHAPS, 1% DTT,
0.5% IPG Buffer pH 3-10. Resuspended proteins, approximately 300
μg, were absorbed overnight into 13 cm Immobilized Dry strip pH
3-10 (GE Healthcare) and focused at 50 μA per strip according to
the manufacturer’s protocol, for a total of 78,300 Vh in an IPGphor
isoelectrofocusing unit (GE Healthcare). After focusing, strips were
equilibrated with equilibration buffer (6 M Urea, 75 mM Tris HCl,
pH 8.8, 29.3% (v/v), 30% glycerol, 2% (w/v) SDS and 0.002% (w/v)
Bromophenol blue containing 10 mg/ml supplemented with 1% DTT
for 15 min, and then with 2.5% iodoacetamide for 15 min on roller
mixer.
The Second Dimension (SDS-PAGE) [23] run was performed
using 12.5% resolving gels with 6% (w/v) stacking gel on a Protean-2
cell (Bio-Rad). The gels were stained with Coomassie Brilliant
Blue (CBB), and images were acquired by Image Scanner LabScan
6.0 (GE Healthcare Biosciences) Proteomic Lab, IITB, India.
The quantification of protein was estimated using the Bradford
Method and standardized with BSA (Bovine Serum Albumin). The
quantification of protein extracts observed low concentrations across
individual replicate samples in each seasons.
1-D SDS-PAGE gels were used to examine the range of protein
MW and to assess the presence of interfering substances in the
muscle protein extracts. The standard of protein determined from
absorbance at 595nm wavelength plotted against Bovine Serum
Album (BSA) concentrations.
In-Gel Digestion
In-gel digestion using trypsin was performed according
to Shevchenko, et al. [26]. The polyacrylamide gel was washed
thoroughly with 100 mM NH4
HCO3
. The protein bands were then
excised from the gel.
Matrix Laser Desorption Ionization Time of Flight/ Time
Of Flight Mass Spectrometry (MALDI-TOF/TOF-MS)
A total of 8 gel spots were excised from the 2D gels, for MALDI-
TOF/TOF-MS analysis; gel spots were picked up from one freshly run
gel. The gel spots were destained and digested overnight with trypsin.
The resulting peptides were extracted following standard techniques
[27]. Peptide fragment mass spectra were acquired in data dependent
Bruker Daltonic Flex analysis with a scan range of 500–3500 m/z;
three averages and up to three precursor ions were selected from the
MS scan (100–3500 m/z). Protein mass score is -10*Log (P), where P
is the probability that the observed match is a random event. Protein
scores greater than 46 were significant (p < 0.05).
Sequence database search
The MS/MS data were subjected to Mascot protein database
search engine (www.matrixscience.com) [28]. The search engine
contains the calculated spectra for all peptides in the National Centre
for Biotechnology Information (NCBI) non-redundant sequences
database (ncbi.nlm.nih.gov) and UniProt (uniprot.org/).
Isobaric tag relative and absolute quantitation (iTRAQ)
labeling
Peptide fractionation was performed using a strong cation
exchange column as described by Zhang, et al. [29]. Peptides of the C.
nigrodigitatus from Ologe and Badagry lagoon were labeled with the
114, 115, 116, and 117 iTRAQ reporters, respectively. The fractioned
peptide samples were dried, while the columns prepared were
activated with 1 ml of Acetonitrile (ACN), 0.1% Formic acid and a
combined 50% (1 ml ACN) and 50% (0.1% FA) and then washed with
0.1% FA. iTRAQ was performed using Acquisition Mode AutoMS2
with MS Range: 300-3000 m/z MS Q-TOF with ModelG6550A
(Agilent Technologies). The 4-plex iTRAQ is a reagent used to label
all protein samples in the muscle of C. nigrodigitatus from Ologe and
Badagry Lagoons. A total of 12 representative protein samples for
the 4-plex iTRAQ analysis of C. nigrodigitatus muscles from Ologe
and Badagry Lagoon was quantified, pooled together, fractionated
and analyzed to identify the peptide sequence in the tandem mass
spectrometry.
RESULT AND DISCUSSION
Water quality characteristics
The mean variations of physico-chemical properties of water
quality of the lagoons are presented in table 1. The water quality
parameters indicates significant differences (p < 0.05) across sampling
period in each lagoons. Furthermore, T-Test revealed significant
differences (p < 0.05) in pH, Salinity, Dissolved Oxygen (DO) and
Biochemical Oxygen Demand (BOD) between the two Lagoons
across the sampling period. The DO and BOD levels in the water fell
below permissible limit of FMEnv (2003).
Table 2 showed the mean levels of heavy metals in sediment
at Ologe and Badagry lagoons. ANOVA revealed that all metals in
the sediment, except Ni showed significant differences (p < 0.05) in
both lagoons. The T-test also indicated that there was a significant
difference (p < 0.05) in the metals of both lagoons. Furthermore,
Table 1: Mean variations of the water quality parameters in Ologe and Badagry
Lagoons.
S/N PARAMETERS OLOGE BADAGRY
T-test (p
< 0.05)
FMEnv
(2003)
1
Water Temperature
(o
C)
29.82 ± 0.19ab
30.75 ± 0.23ab
0.103 < 40
2 pH 6.83 ± 0.11a
8.30 ± 0.06ab
0.000 6.0 - 9.0
3 Conductivity (μS/m) 188.16 ± 2.97bc
190.13 ± 3.52a
0.433 -
4 Salinity (%) 3.85 ± 0.27 5.20 ± 0.22 0.015 -
5
Dissolved Oxygen
(DO) (mg/L)
3.89 ± 0.18abc
5.75 ± 0.13bc
0.002 5
6
Biochemical Oxygen
Demand (mg/L)
74.11 ± 0.88bc
37.54 ± 3.46a
0.000 10
7 Rainfall (mm) 231.57 ± 43.77 231.57 ± 43.77
Alphabets with the letter "a" indicate no significantly different means (p > 0.05).
FMEnv. (2003)-Nigerian Water Quality Standard for Inland Surface Water.
Table 2: Mean concentration of Heavy metals in sediment at Ologe and
Badagry lagoons.
Heavy
metals
(mg / kg)
OLOGE LAGOON BADAGRY LAGOON
T-test (p <
0.05)Mean ± S.E.M Range
Mean ±
S.E.M
Range
Cr 30.13 ± 0.72 28.9 - 31.4 24.73 ± 1.29 22.5 - 27.0 0.011
Cd 0.60 ± 0.06 0.47 - 0.70 0.10 ± 0.00 0 - 0.10 0.013
Fe 568.4 ± 0.35 567.8 - 569 623.6 ± 9.93
606.4 -
640.8
0.029
Ni 12.07 ± 2.57 7.6 - 16.5 5.80 ± 0.40 5.1 - 6.5 0.17
Cu 4.10 ± 0.01 0.67 - 4.28 1.30 ± 0.12 1.1 - 1.5 0.002
Zn 100.17 ± 5.46 90.7 - 109.6 48.13 ± 1.41 45.7 - 50.6 0.017
Pb 17.57 ± 0.38 16.9 - 18.2 8.60 ± 0.46 7.8 - 9.4 0.00
S.E.M - Standard Error Mean; T-test at level of p < 0.05 (Ologe and Badagry
lagoons).
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Pearson correlation coefficient (Table 4) showed significant
correlation between environmental variables and protein markers
(over-expression: Up-regulated and suppressed: down-regulated
proteins).
Protein identification by Matrix-Assisted Laser Desorption/
Ionization Time of Flight/ Time of Flight Mass Spectrometry
(MALDI-TOF/TOF-MS)
The results of the analysis on Matrix-Assisted Laser Desorption/
Ionization Time of flight/ Time of flight mass spectrometry (MALDI-
TOF/TOF-MS) carried out for the detection, identification and
characterization of protein markers. From the 2D gel run for spot
picking for MALDI-TOF/TOF-MS, 8 spots were visualized on the
CBB-stained gels of the muscle protein extract of C. nigrodigitatus
(plate 1). The identified 8 protein spots were subjected to Mascot
search engine database for fish species. Of the eight, 2 protein spots
(1 and 4) were involved in cell structure. The MS/MS data (peptide
spectra) subjected to Mascot protein database search engine (www.
matrixscience.com) also identified proteins with mass score greater
than 46 which indicates that there was a significant difference (p <
0.05) and that they were over-expressed (up-regulated). On the other
hand, protein mass score below 46 indicated no significant difference
(p > 0.05) and were down-regulated. At Badagry lagoon, the spectra
and gel map had protein mass scores below the required threshold
level of 46. Spectra are displayed in Figured 5, 6, 7, 8, 9 and 10 had no
significant protein mass scores.
Further analysis indicated that, 2 proteins spots represented by
cytoskeletal proteins, Alpha Actin and Actin cytoplasmic 1 (spots 1
and4)weresignificantlydifferent(p<0.05)indicatingover-expression
in the muscle protein (Plate 1). On the other hand, Fructose-
bisphosphate aldolase A (spot 3), Spot 2 and 8: Metallothionein, spot
5: Parvalbumin beta 1, Spot 6: Hemoglobin subunit beta-1, and Spot
7: Prolactin-1 showed no significant difference (p > 0.05) in muscle
of C. nigrodigitatus indicating that the proteins from spot 2, 3, 5,
6, 7 and 8 were down-regulated in the muscle (Figure 10-15). The
peptide spectra of the cytoskeletal proteins of spot 1 and 4 that are
over-expressed are presented in figure 3 & 4.
erutarepmeT
pH
Conducvity
Salinity
BOD
DO
Rainfall
Cr
Cd
Fe
Ni
Cu
Zn
Pb
MMD_Up
CytOx_Up
RegC_Up
TuB1_Up
LactateD_Up
keran_Up
troponin_Up
Myozenin_Up
RetX_Up
fActCap_Up
SarRet_Up
antrypsin_Up
G6Pi_Dn
LipB_Dn
myosinR_Dn
cytB_Dn
Vam6_Dn
aCryB_Dn
TPI_Dn
Vasa_Dn
CMMH_Dn
Myosin7_Dn
Temperature 1
pH 0.168 1
Conduc vity -.982* -0.349 1
Salinity -0.197 0.929 0.012 1
BOD 0.759 0.691 -0.853 0.436 1
DO 0.163 .998** -0.344 0.934 0.704 1
Rainfall -.996** -0.134 .972* 0.237 -0.709 -0.124 1
Cr -0.828 -0.409 0.865 -0.149 -0.938 -0.432 0.773 1
Cd -0.936 -0.505 .985* -0.16 -0.912 -0.5 0.92 0.872 1
Fe 0.728 0.669 -0.818 0.362 0.744 0.643 -0.741 -0.558 -0.877 1
Ni -.982* -0.016 0.937 0.351 -0.625 -0.005 .993** 0.718 0.866 -0.676 1
Cu -0.8 -0.717 0.896 -0.41 -0.914 -0.707 0.786 0.776 .957* -.951* 0.708 1
Zn -0.925 -0.524 .979* -0.177 -0.896 -0.516 0.914 0.837 .998** -0.907 0.859 .968* 1
Pb -0.734 -0.782 0.845 -0.496 -0.9 -0.771 0.719 0.73 0.922 -.955* 0.634 .995** 0.937 1
MMD_Up 0.544 -0.68 -0.388 -0.898 -0.129 -0.699 -0.594 -0.087 -0.233 0.084 -0.686 -0.003 -0.229 0.086 1
CytOx_Up -0.886 -0.55 0.947 -0.202 -0.837 -0.533 0.888 0.738 .973* -.960* 0.834 .972* .986* 0.95 -0.231 1
RegC_Up -0.802 0.454 0.677 0.741 -0.267 0.458 0.819 0.5 0.54 -0.252 0.878 0.289 0.518 0.189 -0.903 0.468 1
TuB1_Up -.965* 0.089 0.901 0.433 -0.606 0.09 .965* 0.758 0.814 -0.538 .980* 0.616 0.794 0.531 -0.706 0.739 0.926 1
LactateD_Up -0.513 -0.889 0.656 -0.67 -0.771 -0.871 0.507 0.517 0.767 -0.934 0.411 0.919 0.797 .954* 0.277 0.846 -0.075 0.272 1
kera n_Up 0.097 -0.928 0.084 -0.93 -0.391 -0.913 -0.105 0.051 0.248 -0.57 -0.21 0.516 0.287 0.603 0.72 0.364 -0.65 -0.352 0.804 1
troponin_Up -0.59 -0.629 0.68 -0.354 -0.568 -0.593 0.622 0.342 0.742 -.971* 0.565 0.85 0.786 0.867 -0.083 0.875 0.152 0.397 0.903 0.621 1
Myozenin_Up 0.031 -0.878 0.137 -0.921 -0.627 -0.902 -0.1 0.437 0.287 -0.296 -0.215 0.46 0.278 0.521 0.852 0.239 -0.56 -0.231 0.595 0.742 0.197 1
RetX_Up -0.153 -0.482 0.236 -0.501 -0.652 -0.529 0.06 0.68 0.306 -0.016 -0.017 0.308 0.258 0.312 0.545 0.13 -0.154 0.081 0.22 0.193 -0.187 0.797 1
fActCap_Up -0.785 -0.666 0.872 -0.347 -0.813 -0.646 0.789 0.652 0.926 -.993** 0.722 .979* 0.948 .976* -0.093 .983* 0.305 0.602 0.925 0.525 0.935 0.329 0.109 1
SarRet_Up .983* 0.005 -0.935 -0.344 0.681 0.005 -.976* -0.813 -0.862 0.589 -.980* -0.682 -0.843 -0.602 0.634 -0.785 -0.885 -.995** -0.348 0.274 -0.439 0.136 -0.153 -0.657 1
an trypsin_Up-0.735 -0.483 0.79 -0.263 -0.944 -0.511 0.669 .988* 0.816 -0.507 0.603 0.747 0.78 0.712 0.06 0.673 0.371 0.65 0.52 0.125 0.286 0.558 0.78 0.604 -0.714 1
G6Pi_Dn 0.366 -0.849 -0.187 -.984* -0.288 -0.856 -0.406 0.009 -0.016 -0.198 -0.512 0.241 0 0.333 .957* 0.025 -0.844 -0.58 0.532 0.887 0.209 0.891 0.476 0.178 0.498 0.137 1
LipB_Dn -.987* -0.123 .962* 0.251 -0.677 -0.11 .998** 0.73 0.908 -0.757 .993** 0.78 0.907 0.715 -0.617 0.892 0.817 .955* 0.513 -0.094 0.653 -0.138 -0.005 0.798 -.963* 0.621 -0.419 1
myosinR_Dn -0.173 -0.345 0.229 -0.194 -0.025 -0.292 0.242 -0.224 0.275 -0.679 0.224 0.421 0.34 0.457 -0.127 0.49 -0.053 0.031 0.601 0.542 0.835 -0.131 -0.639 0.588 -0.035 -0.288 0.121 0.301 1
cytB_Dn 0.668 -0.415 -0.557 -0.701 0.041 -0.447 -0.728 -0.154 -0.436 0.398 -0.794 -0.27 -0.451 -0.197 0.939 -0.489 -0.855 -0.751 -0.046 0.438 -0.418 0.729 0.597 -0.389 0.697 0 0.8 -0.763 -0.425 1
Vam6_Dn -0.549 -0.682 0.651 -0.423 -0.578 -0.647 0.577 0.334 0.725 -.967* 0.513 0.852 0.77 0.875 -0.007 0.859 0.083 0.344 0.927 0.678 .997** 0.261 -0.148 0.93 -0.39 0.289 0.282 0.607 0.83 -0.348 1
aCryB_Dn -0.842 -0.529 0.901 -0.184 -0.75 -0.504 0.858 0.626 0.927 -.980* 0.809 0.944 .950* 0.927 -0.262 .988* 0.441 0.692 0.852 0.395 0.93 0.163 -0.011 .985* -0.731 0.552 0.011 0.872 0.614 -0.543 0.913 1
TPI_Dn -0.624 -0.784 0.741 -0.583 -.982* -0.801 0.565 0.886 0.827 -0.688 0.468 0.87 0.811 0.872 0.313 0.752 0.088 0.448 0.782 0.501 0.517 0.762 0.736 0.753 -0.534 0.921 0.453 0.529 -0.014 0.143 0.54 0.661 1
Vasa_Dn -0.61 -0.736 0.72 -0.465 -0.687 -0.707 0.626 0.456 0.798 -.987* 0.554 0.915 0.835 0.935 0.03 0.905 0.106 0.399 .962* 0.682 .985* 0.352 -0.014 .966* -0.455 0.419 0.315 0.647 0.743 -0.303 .990* 0.938 0.653 1
CMMH_Dn 0.489 -0.762 -0.321 -0.948 -0.169 -0.773 -0.53 -0.09 -0.156 -0.054 -0.628 0.098 -0.142 0.193 .986* -0.121 -0.904 -0.68 0.403 0.824 0.074 0.855 0.464 0.032 0.605 0.047 .989* -0.545 0.038 0.87 0.149 -0.135 0.346 0.176 1
Myosin7_Dn 0.494 -0.577 -0.361 -0.701 0.165 -0.548 -0.466 -0.497 -0.226 -0.235 -0.525 0.056 -0.171 0.154 0.623 -0.043 -0.796 -0.68 0.442 0.839 0.403 0.369 -0.25 0.141 0.644 -0.436 0.732 -0.431 0.646 0.402 0.453 0.051 -0.051 0.385 0.725 1
*Correlation is significant at the 0.05 level (2-tailed)
**Correlation is significant at the 0.01 level (2-tailed)
Figure 2: iTRAQ Analytic process of the muscle of C. nigrodigitatus in OL-
Ologe and BL-Badagry lagoons.
Table 4: Pearson correlation coefficient of physico-chemical properties of the water, heavy metals in sediment and up and down regulated proteins in C.
gariepinus from Ologe and Badagry lagoons.
8. International Journal of Proteomics & Bioinformatics
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Protein spot 1
Figure 3: Multiple spectra of peptides map 1 of C. nigrodigitatus muscle
(MALDI TOF/ TOF Analysis) at Ologe Lagoon. Peptides map with spectra
indicated significant differences (p < 0.05).
Protein spot 4
Figure 4: Multiple spectra of peptides map 4 of C. nigrodigitatus muscle
(MALDI TOF/ TOF Analysis) at Ologe Lagoon. Peptides map with spectra
indicated significant differences (p < 0.05).
Figure 5: Multiple spectra of peptides map 2 of C. nigrodigitatus muscle
(MALDI TOF/ TOF Analysis) at Ologe Lagoon. Peptides map with spectra
indicated no significant differences (p > 0.05).
Protein identification by isobaric Tag for Relative and Absolute
Quantitation (iTRAQ)
The measurement of protein responses to contaminants in
muscle tissue of C. nigrodigitatus using iTRAQ detected 116 peptide
sequences. The peptide sequences after subjection to tandem Mass
Spectrometry/ Mass Spectrometry in a pool across sample stations
and seasons revealed the identity of potential protein markers in
muscles of C. nigrodigitatus from Ologe and Badagry lagoons. Further
evaluation of the 116 identified peptide sequences subjected to the
National Centre for Biotechnology Information (NCBI) database
provided little or no information on the protein sequence for
Chrysichthys nigrodigitatus in the database. However, on evaluation Figure 6: Multiple spectra of peptides map 3 of C. nigrodigitatus muscle
(MALDI TOF/ TOF Analysis) at Ologe Lagoon. Peptides map with spectra
indicated no significant differences (p > 0.05).
Figure 7: Multiple spectra of peptides of C. nigrodigitatus muscle (MALDI
TOF/ TOF Analysis) at Badagry Lagoon. Peptides spectra indicated no
significant differences (p > 0.05).
Figure 8: Multiple spectra of peptides of C. nigrodigitatus muscle (MALDI
TOF/ TOF Analysis) at Badagry Lagoon. Peptides spectra indicated no
significant differences (p > 0.05).
using UniProt database search engine, the gene identity of the protein
markers was revealed based on their relationship with the molecular
weight and gene expressions from other related catfishes (Table 3).
A total of 100 (86.20%) proteins were observed to be over-
expressed (up-regulated) in the muscle of C. nigrodigitatus as a
consequent of thermal stress and hypoxic conditions influenced
by various environmental factors representing 60.34% and 25.86%
in the muscle of C. nigrodigitatus from Ologe and Badagry lagoons
respectively.
The results also indicated that, of the 116 identified proteins in
9. International Journal of Proteomics & Bioinformatics
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the muscle of C. nigrodigitatus 70 proteins were up-regulated and
25 proteins down-regulated in Ologe lagoon, while 30 proteins were
over-expressed and 17 proteins were down-regulated in Badagry
lagoon.
The observed up-regulated proteins were identified as; Heat shock
protein- Hsp 70 and 90, Lactate Dehydrogenase A-chain (LDH),
Creatine Kinase (CK), cytochrome c oxidase, regucalcin. Similarly,
total of 47 (40.51%) proteins were observed to be depressed (down-
regulated) in the muscle of C. nigrodigitatus representing 53.19% and
36.17% in the muscle of C. nigrodigitatus from Ologe and Badagry
lagoons respectively. The observed down-regulated proteins were
identified as cytochrome b, Triosephosphate, alpha-β crystalline, and
Tropomyosin isoforms as presented in table 3.
Functional groups and biological pathways of the
identified proteins in the muscle of C. nigrodigitatus
The identified proteins were assigned to several functional
groups with respect to their involvement with cell structures, signal
transduction, enzyme regulation, and oxidative stress as follows: alpha
actinin, alanine aminotransferase, heat shock protein HSP 90-alpha 1,
transmembrane, and β-tubulin, among others as presented in figure
11.
The identified proteins were divided into ten categories,
viz. cytoskeletal proteins, carbohydrate metabolism, nucleotide
metabolism, defense response to oxidative stress, response to stimulus
(Heat shock protein), multicellular organismal development, signal
transduction, electron transport chain, lipid metabolism, and others.
Figure 9: Multiple spectra of peptides of C. nigrodigitatus muscle (MALDI
TOF/ TOF Analysis) at Badagry Lagoon. Peptides spectra indicated no
significant differences (p > 0.05).
Figure 10: Multiple spectra of peptides of C. nigrodigitatus muscle (MALDI
TOF/ TOF Analysis) at Badagry Lagoon. Peptides spectra indicated no
significant differences (p > 0.05).
Stress response
3%
Oxidative stress
response
1%
Musculo-skeletal
system
32%
Carbohydrate
metabolism
16%electron transport
chain
4%
Lipid Metabolism
3%
Nucleic Acid
Metabolism
3%
Signal Transduction
3%
Multicellular
organismal
development
3%
others
32%
Figure 11: Classification of protein from muscle proteome of Chrysichthys
nigrodigitatus. 116 proteins identified by iTRAQ Q-MS/TOF was classified
according to biological processes are based on a UniProt KB search
Cellular components
23.86%
Biological process
32.49%
Molecular function
43.65%
Figure 12: Gene Ontology of the muscle proteome of C. nigrodigitatus in
Ologe and Badagry lagoon
The results of the gene ontology analyses of identified proteins in
the muscle tissue of C. nigrodigitatus also showed that 32.49% were
involved in biological processes, 43.65% in molecular functions and
23.86% in cellular components. Of the 32.49% proteins involved in
biological processes, 32% were associated with musculo-skeletal
system, 16% accounting for carbohydrate metabolism, responses to
stimuli accounted for 3% and 1% of the proteins related to oxidative
stress response as presented in figure 12.
DISCUSSION
The proteomics biomarker of environmental pollution in
Chrysichthys nigrodigitatus was evaluated at Ologe and Badagry
Lagoon between Oct., 2013 and Apr., 2015. Proteomics, the global
analysis of protein synthesis, studies the protein expression patterns
in response to environmental change. Two-dimensional protein
gels, combined with peptide mass mapping by MALDI TOF MS for
protein identification, as well as iTRAQ was used for determining
differential protein synthesis in biological systems.
The results of the 2-D gel electrophoresis and MALDI-TOF/TOF
MS carried out to investigate the proteome composition and identify
protein expression patterns revealed a total of 8 individual spots on
the basis of their Peptide Mass Fingerprints (PMF). This represented 2
proteins peptide identified as α-Actin and Actin cytoplasmic 1. These
protein markers was also observed to be over-expressed, indicating
the attempt of maintaining cell shape and functionality in cases of
hypoxic and heat shock stress.
The proteomic iTRAQ analysis, a 4-plex labeled sample from
Ologe and Badagry Lagoons identified and quantified 116 protein
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markers. A total of 70 up-regulated and 25 down-regulated proteins
in the muscles of C. nigrodigitatus from Ologe Lagoon and 30 up-
regulated and 17 down-regulated proteins in the muscle of C.
nigrodigitatus from Badagry Lagoon. This is an indication of changes
in the protein expressions (depressed and over-expressed) in the
muscle of C. nigrodigitatus induced by oxidative stress, heat shock
and other variables of environmental stress in an environmentally
polluted Lagoon. Adaptation with the environmental stress leads
to elevated expression of heat shock genes that produce heat shock
proteins, which in turn interact with stress-denatured proteins to
maintain or restore their native structures and prevent aggregation
and degradation [30,31].
The α-tubulin proteins are linked to heavy metal tolerance
[32,33], which was exemplified in this study where α-tubulin 1 was
significantly enhanced in the muscle with an increase protein fold
(over-expressed) in response to the heavy metal in the lagoons.
Keratins are important intermediate filament proteins, and their
primary function is to protect the cells from stress damage that
may result in cell death [34,35]. The up-regulation of keratins in the
muscle of C. nigrodigitatus was expected to fight against oxidative
stress induction from both lagoons.
Actins are highly conserved proteins that are involved in various
important cellular processes including cell motility, cell signaling,
and the establishment and maintenance of cell junctions and cell
shape. The change of actin isoforms (α- and β-Actin) in abundance
might have been related to induce oxidative stress in Ologe and
Badagry lagoons, since Actins can be a direct target for oxidative
modification [36]. Actin is a cytoskeletal protein that is ubiquitously
expressed in many eukaryotic cells and functions as maintenance of
the cytoskeleton, cell motility and muscle contraction. The changes
in the level of cytoskeletal and structural proteins expression can be
very often attributed to the attempt of maintaining cell shape and
functionality in cases of hypoxic [37,38] associated with low dissolved
oxygen and slightly acidic pH level of the water quality, which is
indication of an environmentally stressed Ologe lagoon, as compared
to Badagry Lagoon indicating no significant difference (p > 0.05) of
the protein markers in the muscle of C. nigrodigitatus.
The Metallothionein (MT) was identified to be down-regulated
in the muscle of C. nigrodigitatus. MT is a ubiquitous, low molecular
weight, cysteine-rich (>30%) protein that avidly binds various
transition elements. Metallothionein is an excellent environmental
biomarker. Stress caused by both bio-active and non-bioactive
metals induces a variety of complex changes in fish physiology and
the possible consequent physiological alterations as observed in C.
nigrodigitatus from Ologe and Badagry lagoons.
Another identified protein was Parvalbumin beta 1, involved in
relaxation after contraction of the muscles. It binds two calcium ions
and also involves in calcium signaling. The Parvalbumin was observed
to be down-regulated in the muscle which disrupts the involvement
at the calcium binding site, and relaxation of the muscles which might
be induced by environmental stressors. Prolactin was also identified
to be down-regulated. Prolactin 1 is a protein hormone of the anterior
pituitary gland.
HSP-90 belongs to the Heat Shock Protein Family (HSPs), which
are well known to protect the structure and function of cells from
stress (e.g. metal attack, climate change, thermal stress and hypoxic
conditions generated from other environmental factors) and play an
important role in maintaining cellular homeostasis [39]. The deletion
of HSP-90 is lethal for eukaryotic cells [40], and in this study, the over-
expressed of HSP-90 suggested that the muscle of C. nigrodigitatus
inhibited the stressed condition by enhancing the Hsp in the Ologe
lagoon, while at Badagry lagoon the heat shock protein beta 1 was
observed to be suppressed.
Enolase is a cytosolic enzyme involved in carbohydrate
metabolism, cell differentiation, and normal growth, and a decline of
enolase activity results in abnormal growth and reduced metabolism
in the muscle. The enolase 1 alpha, partial and beta-enolase were up-
regulated in the muscle which indicates an active metabolism with
normal growth of the fish species in the lagoons. Creatine kinase is
an essential enzyme for energy buffering to maintain cellular energy
homeostasis. Its activity is inhibited by metals [41]. The increased
(up-regulation) levels of enolase 1 (alpha) and creatine kinase in the
muscles of C. nigrodigitatus are an adaptive feedback to inhibitory
activity in the muscle under oxidative stress. Cytochrome c oxidase is
the component of the respiratory chain that catalyzes the reduction
of oxygen to water. The over expression of Cytochrome C Oxidase
(COX2) in muscle proteome of C. nigrodigitatus, is an indication
of adaptation to hypoxic condition induced by admixture of
environmental pollutants in the dry season at Ologe lagoon.
Regucalcin was identified to be over-expressed in the muscle of
C. nigrodigitatus in both lagoons. Regucalcin has been found to play
a multifunctional role in different tissues, and is primarily involved
in the maintenance of intracellular Ca2+
homeostasis (calcium
ion binding and enzyme regulator activity). Over-expression of
regucalcin are usually found to enhance glucose utilization and lipid
production in fish when exposed to severe oxidative stress observed
in Ologe and Badagry Lagoons respectively. Cytochrome b protein
are component of the ubiquinol - cytochrome c reductase complex,
which functions as respiratory electron transport chain that generates
an electrochemical potential coupled to ATP synthesis. The over-
expression of the cytochrome b (cyt b) indicates a significant decline
of dissolved oxygen, rendering the cyt b to increase its electron
transport chain for respiratory functions as an adaptive mechanism
in the muscle of C. nigrodigitatus at the dry season in Ologe Lagoon.
Another identified protein that was over-expressed is Lactate
Dehydrogenase A-Chain (LDH), is a key enzyme in the control of
energy metabolism, catalyzing the interconversion of pyruvate to
lactate and regulating the levels of these metabolites in accordance
with oxygen availability. Under anaerobic conditions, the isoform
LDH-A4 (isozyme A4) preferentially converts pyruvate to lactate.
This isoform is found predominantly in poorly vascularized tissues
with low partial Pressure of Oxygen (pO2
), such as skeletal muscle in
both lagoons and seasons.
Triosephosphate Isomerase (TPI) was observed to be down-
regulated in the muscle of C. nigrodigitatus. TPI is a glycolytic enzyme
that catalyzes the conversion between glyceraldehyde-3-phosphate
and dihydroxyacetone phosphate; it is essential for efficient energy
production by being involved in several metabolic pathways. The
significant changes of TPI expression might alter catalytic process of
energy production induced by pollutants in Ologe Lagoon. Another
identified down-regulated protein was α-B crystalline, which is
responsible for metal ion binding, structural constituents for the
eye lens. The less abundant corneal proteins might be unable to
contribute structurally to its transparency and optical properties of C.
nigrodigitatus at the Badagry Lagoon.
Phosphoglucomutase-1 was to be down-regulated in the muscle
11. International Journal of Proteomics & Bioinformatics
SCIRES Literature - Volume 4 Issue 1 - www.scireslit.com Page - 025
of C. nigrodigitatus in Ologe lagoon. Phosphoglucomutase-1 is a key
enzyme in the metabolism of glycogen and protein glycosylation. It is
responsible for the reversible inter conversion of glucose 1-phosphate
to glucose 6-phosphate, both of which are key intermediates in the
synthesis and breakdown of glycogen and galactose metabolism. It is
also important for the formation of UDP-glucose which is an essential
intermediary metabolite in protein glycosylation. Inhibition of
phosphoglucomutase has drastic effects on carbohydrate metabolism
which reduces the steady-state levels of UDP-glucose, resulting
in a defect of glycogen and trehalose biosynthesis, while galactose
metabolism is inhibited, leading to galactosemia, accumulation of
galactose 1-phosphate and Glucose 1-phosphate i.e., poor glycogen
turnover.
Tropomyosin alpha-1 was observed to be up-regulated in both
lagoons, while Tropomyosin alpha-3 chain was observed to be down-
regulated as well in both lagoons and Tropomyosin isoforms have
been reported both in condition of thermal stress [37] and hypoxia
[38].
CONCLUSION
This study identified protein markers that are over-expressed
(Up-regulated) and depressed (Down-regulated) in the muscle of
C. nigrodigitatus which serves as prognostic tools (MALDI-TOF/
TOF and iTRAQ approach) to assess the pollution status of Ologe
and Badagry lagoons, and useful for biotechnological interventions
in fish health and disease management; besides adding to the
existing knowledge base on comparative muscle proteomics on C.
nigrodigitatus of a tropical ecosystem.
ACKNOWLEDGEMENT
The author appreciate the mentorship and support of Prof.
Chukwu, L. O. of Department of Marine Sciences, University of
Lagos, Prof. Sanjeeva S. and all Ph.D. colleagues of department of
Bioengineering and Bioinformatics, Proteomics unit, Indian Institute
of Technology Bombay (IITB), Powai Mumbai India.
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