This study investigated the effects of varied doses of nicotine (0.2 mg/100g, 0.4 mg/100g, 0.6 mg/100g) on morphometric parameters of the testis in male albino Wistar rats over 30 days. The parameters measured included testicular weight, seminiferous tubule diameter, height of epithelium, sertoli-germ cell ratio, and Johnsen's score. Nicotine caused a significant dose-dependent reduction in most parameters, indicating it adversely affected spermatogenesis. Specifically, nicotine significantly reduced the height of epithelium at all doses, and significantly reduced seminiferous tubule diameter, Johnsen's score, and sertoli
Gonadal histo morphology and antifertility effects of bonny light crude oil i...Alexander Decker
This document summarizes a study that investigated the effects of ingesting Bonny Light Crude Oil (BLCO) on male fertility in rats. Rats were given varying doses of BLCO for 60 days, then sperm counts were analyzed and testes were examined histologically. Sperm counts significantly decreased in treated rats compared to controls. Histological examination found degenerative and necrotic changes in testes of treated rats, including fewer sperm-producing cells and damage to Sertoli cells that support sperm maturation. The results suggest that ingesting BLCO has adverse effects on the male reproductive system and fertility in rats.
This document describes a method for rapidly detecting whether plant mutants are homozygous or heterozygous using CEL-I endonuclease. CEL-I endonuclease is a mismatch-specific endonuclease that can cleave DNA at sites of mismatches between DNA strands. The method involves isolating genomic DNA from mutant plants, amplifying a region by PCR, and treating the PCR products with CEL-I endonuclease. Digestion products are then analyzed by gel electrophoresis. For homozygous mutants, no cleavage will occur as there are no mismatches. For heterozygous mutants, cleavage will occur at mismatch sites, producing distinctive banding patterns that allow identification of heterozygotes. This method provides a low
This document provides an overview of genetic toxicity testing guidelines. It discusses the history and aims of toxicity studies. Various in vitro and in vivo genetic toxicology tests are described, including tests for gene mutation, chromosomal abnormalities, and primary DNA damage. Key tests covered include the mammalian erythrocyte micronucleus test, mammalian bone marrow chromosomal aberration test, rodent dominant lethal assay, and mouse heritable translocation assay. The principles, procedures, and parameters of these tests are summarized. References on genetic toxicology guidance documents and studies are also provided.
This presentation gives the brief idea of the various guidelines carried out to study the genetic damage to cells when there is a discover of new active molecule.
This document summarizes a study examining the formation and function of the contractile ring involved in polar body extrusion in the surf clam Spisula. The study found that:
1) The metaphase peripheral aster spreads along the egg cortex in an umbrella-like pattern, leaving a microtubule-poor center.
2) During anaphase, the aster disassembles as a cortical F-actin ring forms that matches the location, size, and pattern of the previous aster.
3) Inhibiting F-actin or myosin blocked polar body formation, while disrupting or stabilizing asters prevented proper ring and polar body formation, supporting the hypothesis that aster spreading
1) C. elegans raised in social environments displayed stronger neural activity, as evidenced by more numerous, intense, and larger synapses compared to C. elegans raised in isolation.
2) Behavioral assays also showed C. elegans from social environments had higher response rates to external stimuli and more thrashes per minute than isolated C. elegans.
3) These results support the hypothesis that more social stimulation leads to increased neural plasticity and synaptic strength in C. elegans.
Methimazole affected spermatogenesis and enhanced proliferation of testicular...Jing Zang
This document summarizes a study on the effects of the antithyroid drug methimazole (MMI) on testicular macrophages and spermatogenesis in rats. The study found that treating rats with MMI for 30 days caused a reduction in body weight and seminiferous tubule diameter, and inhibited spermatogenesis. It also observed a significant increase in the number of macrophages in the testicular interstitium, with the highest numbers near Leydig cells. This suggests apoptotic or inflammatory processes increased in the rat testes after MMI exposure. In conclusion, MMI treatment adversely affected testicular macrophages and spermatogenesis in rats, and induced hypothyroidism that can lead to infertility.
Shital Magar presented on in vitro genotoxicity testing based on OECD guidelines. The presentation covered the objectives of genotoxicity testing, introduction to genotoxicity, history, and details of key in vitro tests including bacterial reverse mutation assay, mammalian cell gene mutation tests, mammalian chromosomal aberration test, and mammalian cell micronucleus test. Parameters and limitations of each test were discussed along with examples of software used to analyze genotoxicity results.
Gonadal histo morphology and antifertility effects of bonny light crude oil i...Alexander Decker
This document summarizes a study that investigated the effects of ingesting Bonny Light Crude Oil (BLCO) on male fertility in rats. Rats were given varying doses of BLCO for 60 days, then sperm counts were analyzed and testes were examined histologically. Sperm counts significantly decreased in treated rats compared to controls. Histological examination found degenerative and necrotic changes in testes of treated rats, including fewer sperm-producing cells and damage to Sertoli cells that support sperm maturation. The results suggest that ingesting BLCO has adverse effects on the male reproductive system and fertility in rats.
This document describes a method for rapidly detecting whether plant mutants are homozygous or heterozygous using CEL-I endonuclease. CEL-I endonuclease is a mismatch-specific endonuclease that can cleave DNA at sites of mismatches between DNA strands. The method involves isolating genomic DNA from mutant plants, amplifying a region by PCR, and treating the PCR products with CEL-I endonuclease. Digestion products are then analyzed by gel electrophoresis. For homozygous mutants, no cleavage will occur as there are no mismatches. For heterozygous mutants, cleavage will occur at mismatch sites, producing distinctive banding patterns that allow identification of heterozygotes. This method provides a low
This document provides an overview of genetic toxicity testing guidelines. It discusses the history and aims of toxicity studies. Various in vitro and in vivo genetic toxicology tests are described, including tests for gene mutation, chromosomal abnormalities, and primary DNA damage. Key tests covered include the mammalian erythrocyte micronucleus test, mammalian bone marrow chromosomal aberration test, rodent dominant lethal assay, and mouse heritable translocation assay. The principles, procedures, and parameters of these tests are summarized. References on genetic toxicology guidance documents and studies are also provided.
This presentation gives the brief idea of the various guidelines carried out to study the genetic damage to cells when there is a discover of new active molecule.
This document summarizes a study examining the formation and function of the contractile ring involved in polar body extrusion in the surf clam Spisula. The study found that:
1) The metaphase peripheral aster spreads along the egg cortex in an umbrella-like pattern, leaving a microtubule-poor center.
2) During anaphase, the aster disassembles as a cortical F-actin ring forms that matches the location, size, and pattern of the previous aster.
3) Inhibiting F-actin or myosin blocked polar body formation, while disrupting or stabilizing asters prevented proper ring and polar body formation, supporting the hypothesis that aster spreading
1) C. elegans raised in social environments displayed stronger neural activity, as evidenced by more numerous, intense, and larger synapses compared to C. elegans raised in isolation.
2) Behavioral assays also showed C. elegans from social environments had higher response rates to external stimuli and more thrashes per minute than isolated C. elegans.
3) These results support the hypothesis that more social stimulation leads to increased neural plasticity and synaptic strength in C. elegans.
Methimazole affected spermatogenesis and enhanced proliferation of testicular...Jing Zang
This document summarizes a study on the effects of the antithyroid drug methimazole (MMI) on testicular macrophages and spermatogenesis in rats. The study found that treating rats with MMI for 30 days caused a reduction in body weight and seminiferous tubule diameter, and inhibited spermatogenesis. It also observed a significant increase in the number of macrophages in the testicular interstitium, with the highest numbers near Leydig cells. This suggests apoptotic or inflammatory processes increased in the rat testes after MMI exposure. In conclusion, MMI treatment adversely affected testicular macrophages and spermatogenesis in rats, and induced hypothyroidism that can lead to infertility.
Shital Magar presented on in vitro genotoxicity testing based on OECD guidelines. The presentation covered the objectives of genotoxicity testing, introduction to genotoxicity, history, and details of key in vitro tests including bacterial reverse mutation assay, mammalian cell gene mutation tests, mammalian chromosomal aberration test, and mammalian cell micronucleus test. Parameters and limitations of each test were discussed along with examples of software used to analyze genotoxicity results.
Managing Health and Disease Using Omics and Big DataLaura Berry
Presented at the NGS Tech and Applications Congress: USA. To find out more, visit:
www.global-engage.com
Michael Snyder is a Professor, Chair of Genetics and Director of the Stanford Center for Genomics and Personalized Medicine at Stanford University. In this presentation Michael discusses using omics and big data to predict disease risk and catch early disease onset.
This experiment tested the effects of silencing two genes, pdl-1 and F26A10.1, on the chemosensory abilities of C. elegans. RNAi was used to suppress the genes in C. elegans, and a chemotaxis assay was performed to assess chemosensory function. The results showed that silencing pdl-1 significantly reduced chemosensation, suggesting it plays a role, while F26A10.1 silencing had no effect, indicating it is not involved in chemosensation. Statistical analysis supported the conclusion that pdl-1 affects chemosensory pathways but F26A10.1 does not.
The efficiency of transgenesis by restriction enzyme mediated integration s...Alexander Decker
This document summarizes a study that tested the efficiency of restriction enzyme mediated integration sperm mediated gene transfer (REMI-SMGT) for generating transgenic rabbits. REMI-SMGT uses restriction enzymes to facilitate integration of exogenous DNA into the genome of sperm cells. The study found that while various treatments successfully internalized exogenous DNA into rabbit sperm heads, only 3 out of 14 newborn rabbits were transgenic by PCR. This low efficiency may reflect the inability of recombinant sperm to fertilize eggs rather than a weakness of REMI-SMGT. Comparable results between liposome and DMSO treatments suggest DMSO interacts with cell membranes rather than DNA. The study was the first to evaluate REMI-SMGT in rabbits and modify
Fundamentals Of Genetic Toxicology In The Pharmaceutical Industry Sept 2010TigerTox
Historical and current perspectives on genetic toxicology, with commentary and slides on assay predictivity and shortcomings, regulatory guidance, and high-throughput screens to enhance preclinical drug safety.
Assignment on Limitation of animal experimentationDeepak Kumar
This document discusses 5 key causes for why animal studies do not reliably predict human outcomes: 1) Interspecies differences in disease susceptibility, drug metabolism, and other factors; 2) Stressful laboratory environments and common procedures that alter animal physiology; 3) False positives from chronic high-dose rodent studies that overwhelm natural defenses; 4) Poor methodological quality in many animal experiments lacking randomization, blinding, and other controls; 5) Publication and other biases that skew available data.
This document discusses the Ames test, which is used to determine if chemicals are mutagenic and potentially carcinogenic. It describes how the Ames test works using a histidine-requiring strain of salmonella bacteria that is exposed to test chemicals - if mutations occur that allow bacterial growth, the chemical is considered mutagenic. The document provides background on Bruce Ames who developed the test, and explains the process and applications of the Ames test in determining cancer risk from chemicals, and its use in biotechnology and industry.
Historically, genetic toxicology has been comprised of bacterial and cell based in vitro assays such as the Ames assay (a bacterial mutagenicity assay), Micronucleus and Chromosomal Aberration assays (mammalian cytogenetic assays), and Mouse Lymphoma Assay (in vitro mammalian cell gene mutation assay). These were routinely used for safety evaluation and are still part of the standard core battery. The emergence of new technologies has facilitated the development of in vitro methods for safe and effective drug and chemical testing.
This BioReliance® toxicology services webinar will explore alternative models, including 3D skin models that comply with the EC Scientific Committee on Consumer Safety (SCCS) recommendations. It will also discuss how the 3Rs (Replace, Reduce, Refine) Principle advocates the exploration of such alternative methods while achieving required goals.
In this webinar, you will learn:
• About in vitro alternatives to animal toxicity testing in pharma, chemical, tobacco, and personal care products.
• How the 3Rs (Replace, Reduce, Refine) Principle advocates exploring alternative methods without compromising the required goals.
• Alternatives to comply with the 7th Amendment to the EC Cosmetics Directive.
The document discusses mutagenicity and carcinogenicity of environmental factors. It defines key terms like mutagen, mutagenicity, carcinogen and carcinogenicity. It describes various types of mutagens and carcinogens. It also summarizes different methods to test for mutagenicity and carcinogenicity, including tests on the molecular, gene and chromosomal level like the Ames test, comet assay and micronucleus test. The document provides an overview of the process of carcinogenesis and challenges in evaluating human carcinogenicity.
Zebrafish as an alternative method for determining the embryo toxicity of pla...Noor Zada
The document systematically reviews 12 articles that evaluated the toxicity of various plant products using zebrafish embryos. The plant products tested included caffeine, cannabinoids, nicotine, curcumin, resveratrol, quercetin, rutin, matrine, sophocarpine, and arecoline. The studies assessed endpoints such as mortality, developmental abnormalities, heart rate, locomotor activity, and growth. Many found similar results to mammalian studies, demonstrating zebrafish embryos can be a valid alternative model for toxicity testing of plant products.
Dr Ayman Ewies - Effect of stretch & Levormeloxifene on the cytoskeleton of c...AymanEwies
1) The study examined the effects of stretch and the drug levormeloxifene on the cytoskeleton of fibroblasts from the cardinal ligaments.
2) Fibroblasts exposed to stretch or treated with levormeloxifene showed a higher percentage of abnormal actin configuration, including dissolution and aggregation of actin filaments.
3) Estrogen treatment did not significantly affect actin morphology. The results suggest that stretch and levormeloxifene can disrupt the cytoskeleton of ligament fibroblasts, which may relate to the development of pelvic organ prolapse.
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
This study aimed to develop the brook stickleback as a new model species for assessing endocrine disrupting compounds. The researchers first optimized a qPCR assay to measure expression of the androgen-regulated spiggin gene in kidney tissue and the estrogen-regulated vitellogenin gene in liver tissue. They found basal differences in expression between male and female brook stickleback. Fish were then exposed to model androgens and estrogens, which induced expression of spiggin in female kidney and vitellogenin in male liver, respectively. Exposure also altered organ weights and histopathology. Given the sensitivity of these responses, brook stickleback show potential as a new species for evaluating endocrine
This document discusses various methods for testing the mutagenicity of chemicals, including both prokaryotic and eukaryotic cell systems. It describes the Ames test which uses Salmonella bacteria to identify mutagens, as well as other prokaryotic methods like the host-mediated assay and coliform assay. Eukaryotic methods discussed include the Saccharomyces forward mutation assay, mammalian cell tests, and in vivo assays like the micronucleus test and dominant lethal assay. The document provides details on the procedures and principles of many of these important mutagenicity testing methods.
Genotoxicity studies according to oecd guildline.Diana Lou
This document provides information about genotoxicity studies and various genotoxicity testing methods. It discusses that genotoxicity tests identify compounds that cause genetic damage through DNA damage or interference with the cell cycle. The standard battery of genotoxicity tests includes the Ames test (bacterial reverse mutation assay), in vitro mammalian cell micronucleus assay, in vitro mammalian chromosomal aberration assay, and in vivo mammalian erythrocyte micronucleus test, which detect various types of genetic damage. The document outlines the key principles, procedures, and reporting requirements for each of these standard genotoxicity assays.
This document discusses mutagenicity and carcinogenicity testing that is done during drug discovery and development. It defines mutagenicity as the ability of chemicals to cause permanent changes to DNA, potentially leading to inherited mutations, and carcinogenicity as the ability to cause cancer. Common classes of mutagenic and carcinogenic compounds are described. Methods of mutagenicity testing include the Ames test, which uses Salmonella bacteria to identify mutagens, and other in vitro and in vivo assays. The document emphasizes the importance of screening drugs for these toxic effects before human use to avoid dangerous inherited mutations or cancer risks.
This document summarizes a talk on organ development and the use of skin and hair as a model system. It discusses how organs form through morphometric modules guided by developmental signals. The skin and hair follicle are presented as an advantageous model due to the ability to culture stem cells, track defects in mice, and their dispensability. Key processes in hair follicle development highlighted include cadherin switching regulated by the Wnt and BMP signaling pathways and transcription factors like Lef1 and beta-catenin. Downregulation of E-cadherin, driven by these signals, is implicated in promoting polarity changes required for budding morphogenesis.
This study investigated the antioxidant effects of nebivolol in protecting against testicular damage caused by torsion-detorsion injury in rats. Forty rats were divided into four groups: a control group, a torsion group, a torsion/detorsion group, and a torsion/detorsion+nebivolol group. Biochemical assays and histopathological examination found that torsion-detorsion injury increased oxidative stress markers and apoptosis in testicular tissue, while administration of nebivolol before detorsion decreased oxidative stress and apoptosis. The study suggests that nebivolol has a protective effect against ischemia-reperfusion injury in the testes caused by torsion-
Objective: To identify interstitial cells of Cajal (ICC) in the common bile duct of Kunming mice.
Study Design: Common bile ducts obtained from the Kunming mice were prepared for immunohistochemical investigations using the c-kit antibody. Immunoelectron microscopy was used to detect the expression of c-kit in the ICC of the common bile duct. Transmission electron microscopy showed ultrastructure of ICC in the murine bile duct. Reverse transcription–polymerase chain reaction (RT-PCR) and western blot were used to confirm the expression of mRNA specific for the c-kit gene and production of c-kit protein in the Kunming mice common bile duct.
Results: Immunohistochemistry revealed that ICC in the murine common bile duct are c-kit positive and the ICC are located in the tela submucosa and the tunica muscularis of the murine common bile duct and do not connect with each other. Immunoelectron microscopy confirmed the expression of Kit by ICC in the murine common bile duct. Transmission electron microscopy showed that ICC in the murine common bile duct have long processes, abundant mitochondria, plenty of smooth endoplasmic reticulum (sER), a lot of lysosomes, and dense bodies. The caveolae of ICC are distinctive. At the same time, RT-PCR indicated that the Kunming mice common bile duct expressed mRNA specific for the c-kit gene, and western blot analysis showed the evidence of production of c-kit protein in the Kunming mice common bile duct.
Conclusion: ICC are found in the Kunming mice common bile duct, which is likely to lead to the development of motility study of the common bile duct.
Keywords: common bile duct; electron microscopy; immuno-electron microscopy; interstitial cells of Cajal; intestines; smooth muscle; tyrosine kinase receptor (c-kit)
Ldb 145 Geni Mutanti_2014-11-19 Jamora - dalla ricerca al prodotto 5laboratoridalbasso
1) Fibulin-5 expression is elevated in the dermis of Snail transgenic skin, a model of tissue fibrosis. Deletion of Fibulin-5 decreases tissue stiffness and the amount of elastic fibers but not collagen fibers.
2) Depletion of Fibulin-5 decreases inflammation in the fibrotic skin as measured by reduced macrophage and T cell infiltration.
3) Fibulin-5 promotes the production of inflammatory chemokines from dermal fibroblasts and accentuates their activation both in vivo and in response to substrate stiffness in vitro.
Ld b 145 geni mutanti_2014-11-18 jamora - ricerca scientifica 3laboratoridalbasso
This document discusses the role of the transcription factor Snail in wound healing and fibrosis. It finds that overexpression of Snail in the epidermis of mice induces dermal fibrosis through increased secretion of spondin-2/mindin by keratinocytes. Spondin-2 then activates dermal fibroblasts by stimulating the NF-kB pathway, leading to increased expression of genes associated with myofibroblast differentiation and extracellular matrix production. This establishes a paracrine signaling mechanism from epidermal Snail to induce dermal fibrosis through spondin-2.
Spermatotoxic impact of bonny light crude oil (BLCO) ingestion on adult male ...lukeman Joseph Ade shittu
Increasing concern has been expressed about the possible declining trend in the sperm quality and sperm count of man as a result of exposure to environmental estrogenic agents in the past few years now. There is a general paucity of knowledge of BLCO ingestion on the reproductive effect. Hence, we aim to evaluate the impact of sub-lethal dose of BLCO ingestion on semen parameters of adult male mice. Initial acute toxicity study was carried out to determine the lethal dose of BLCO, which was calculated to be 37.4 mg/Kg body wt. A sub-lethal dose of 20 mg/Kg bwt /day of BLCO were then given to 8 male mice in the experimental group. While, the control group of 7 animals received equal volume of 0.9% normal saline via oral garvage for 2 weeks. Data were analysed using SPSS 12 statistical software with P < 0.05 considered statistically significant. There was a significant (P < 0.05) weight gain in the treated group with a significant (P < 0.05) reduction in sperm motility in the treated compared with control. The sperm density of treated and control were 14.5 x 106 /ml and 20.5 x 106 /ml respectively. However, there were also no significant difference in the relative testicular weight and sperm density of treated from that of the control respectively. Thus, it was concluded that BLCO ingestion is spermatotoxic in the adult male Swiss mice
This study investigated the effects of gallic acid on testicular injury caused by ischemia-reperfusion in a rat testicular torsion model. Forty rats were divided into four groups: a control group, a torsion group, a torsion/detorsion group, and a torsion/detorsion plus gallic acid group. Biochemical markers and immunohistochemical staining for caspase-3 and TNF-α were analyzed. The results showed that gallic acid treatment decreased oxidative stress markers, reduced apoptosis and inflammation, and helped protect testicular tissue compared to the torsion/detorsion group without treatment. The study suggests that gallic acid may be a potential therapeutic agent for testicular ischemia-reperfusion injury.
Managing Health and Disease Using Omics and Big DataLaura Berry
Presented at the NGS Tech and Applications Congress: USA. To find out more, visit:
www.global-engage.com
Michael Snyder is a Professor, Chair of Genetics and Director of the Stanford Center for Genomics and Personalized Medicine at Stanford University. In this presentation Michael discusses using omics and big data to predict disease risk and catch early disease onset.
This experiment tested the effects of silencing two genes, pdl-1 and F26A10.1, on the chemosensory abilities of C. elegans. RNAi was used to suppress the genes in C. elegans, and a chemotaxis assay was performed to assess chemosensory function. The results showed that silencing pdl-1 significantly reduced chemosensation, suggesting it plays a role, while F26A10.1 silencing had no effect, indicating it is not involved in chemosensation. Statistical analysis supported the conclusion that pdl-1 affects chemosensory pathways but F26A10.1 does not.
The efficiency of transgenesis by restriction enzyme mediated integration s...Alexander Decker
This document summarizes a study that tested the efficiency of restriction enzyme mediated integration sperm mediated gene transfer (REMI-SMGT) for generating transgenic rabbits. REMI-SMGT uses restriction enzymes to facilitate integration of exogenous DNA into the genome of sperm cells. The study found that while various treatments successfully internalized exogenous DNA into rabbit sperm heads, only 3 out of 14 newborn rabbits were transgenic by PCR. This low efficiency may reflect the inability of recombinant sperm to fertilize eggs rather than a weakness of REMI-SMGT. Comparable results between liposome and DMSO treatments suggest DMSO interacts with cell membranes rather than DNA. The study was the first to evaluate REMI-SMGT in rabbits and modify
Fundamentals Of Genetic Toxicology In The Pharmaceutical Industry Sept 2010TigerTox
Historical and current perspectives on genetic toxicology, with commentary and slides on assay predictivity and shortcomings, regulatory guidance, and high-throughput screens to enhance preclinical drug safety.
Assignment on Limitation of animal experimentationDeepak Kumar
This document discusses 5 key causes for why animal studies do not reliably predict human outcomes: 1) Interspecies differences in disease susceptibility, drug metabolism, and other factors; 2) Stressful laboratory environments and common procedures that alter animal physiology; 3) False positives from chronic high-dose rodent studies that overwhelm natural defenses; 4) Poor methodological quality in many animal experiments lacking randomization, blinding, and other controls; 5) Publication and other biases that skew available data.
This document discusses the Ames test, which is used to determine if chemicals are mutagenic and potentially carcinogenic. It describes how the Ames test works using a histidine-requiring strain of salmonella bacteria that is exposed to test chemicals - if mutations occur that allow bacterial growth, the chemical is considered mutagenic. The document provides background on Bruce Ames who developed the test, and explains the process and applications of the Ames test in determining cancer risk from chemicals, and its use in biotechnology and industry.
Historically, genetic toxicology has been comprised of bacterial and cell based in vitro assays such as the Ames assay (a bacterial mutagenicity assay), Micronucleus and Chromosomal Aberration assays (mammalian cytogenetic assays), and Mouse Lymphoma Assay (in vitro mammalian cell gene mutation assay). These were routinely used for safety evaluation and are still part of the standard core battery. The emergence of new technologies has facilitated the development of in vitro methods for safe and effective drug and chemical testing.
This BioReliance® toxicology services webinar will explore alternative models, including 3D skin models that comply with the EC Scientific Committee on Consumer Safety (SCCS) recommendations. It will also discuss how the 3Rs (Replace, Reduce, Refine) Principle advocates the exploration of such alternative methods while achieving required goals.
In this webinar, you will learn:
• About in vitro alternatives to animal toxicity testing in pharma, chemical, tobacco, and personal care products.
• How the 3Rs (Replace, Reduce, Refine) Principle advocates exploring alternative methods without compromising the required goals.
• Alternatives to comply with the 7th Amendment to the EC Cosmetics Directive.
The document discusses mutagenicity and carcinogenicity of environmental factors. It defines key terms like mutagen, mutagenicity, carcinogen and carcinogenicity. It describes various types of mutagens and carcinogens. It also summarizes different methods to test for mutagenicity and carcinogenicity, including tests on the molecular, gene and chromosomal level like the Ames test, comet assay and micronucleus test. The document provides an overview of the process of carcinogenesis and challenges in evaluating human carcinogenicity.
Zebrafish as an alternative method for determining the embryo toxicity of pla...Noor Zada
The document systematically reviews 12 articles that evaluated the toxicity of various plant products using zebrafish embryos. The plant products tested included caffeine, cannabinoids, nicotine, curcumin, resveratrol, quercetin, rutin, matrine, sophocarpine, and arecoline. The studies assessed endpoints such as mortality, developmental abnormalities, heart rate, locomotor activity, and growth. Many found similar results to mammalian studies, demonstrating zebrafish embryos can be a valid alternative model for toxicity testing of plant products.
Dr Ayman Ewies - Effect of stretch & Levormeloxifene on the cytoskeleton of c...AymanEwies
1) The study examined the effects of stretch and the drug levormeloxifene on the cytoskeleton of fibroblasts from the cardinal ligaments.
2) Fibroblasts exposed to stretch or treated with levormeloxifene showed a higher percentage of abnormal actin configuration, including dissolution and aggregation of actin filaments.
3) Estrogen treatment did not significantly affect actin morphology. The results suggest that stretch and levormeloxifene can disrupt the cytoskeleton of ligament fibroblasts, which may relate to the development of pelvic organ prolapse.
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
This study aimed to develop the brook stickleback as a new model species for assessing endocrine disrupting compounds. The researchers first optimized a qPCR assay to measure expression of the androgen-regulated spiggin gene in kidney tissue and the estrogen-regulated vitellogenin gene in liver tissue. They found basal differences in expression between male and female brook stickleback. Fish were then exposed to model androgens and estrogens, which induced expression of spiggin in female kidney and vitellogenin in male liver, respectively. Exposure also altered organ weights and histopathology. Given the sensitivity of these responses, brook stickleback show potential as a new species for evaluating endocrine
This document discusses various methods for testing the mutagenicity of chemicals, including both prokaryotic and eukaryotic cell systems. It describes the Ames test which uses Salmonella bacteria to identify mutagens, as well as other prokaryotic methods like the host-mediated assay and coliform assay. Eukaryotic methods discussed include the Saccharomyces forward mutation assay, mammalian cell tests, and in vivo assays like the micronucleus test and dominant lethal assay. The document provides details on the procedures and principles of many of these important mutagenicity testing methods.
Genotoxicity studies according to oecd guildline.Diana Lou
This document provides information about genotoxicity studies and various genotoxicity testing methods. It discusses that genotoxicity tests identify compounds that cause genetic damage through DNA damage or interference with the cell cycle. The standard battery of genotoxicity tests includes the Ames test (bacterial reverse mutation assay), in vitro mammalian cell micronucleus assay, in vitro mammalian chromosomal aberration assay, and in vivo mammalian erythrocyte micronucleus test, which detect various types of genetic damage. The document outlines the key principles, procedures, and reporting requirements for each of these standard genotoxicity assays.
This document discusses mutagenicity and carcinogenicity testing that is done during drug discovery and development. It defines mutagenicity as the ability of chemicals to cause permanent changes to DNA, potentially leading to inherited mutations, and carcinogenicity as the ability to cause cancer. Common classes of mutagenic and carcinogenic compounds are described. Methods of mutagenicity testing include the Ames test, which uses Salmonella bacteria to identify mutagens, and other in vitro and in vivo assays. The document emphasizes the importance of screening drugs for these toxic effects before human use to avoid dangerous inherited mutations or cancer risks.
This document summarizes a talk on organ development and the use of skin and hair as a model system. It discusses how organs form through morphometric modules guided by developmental signals. The skin and hair follicle are presented as an advantageous model due to the ability to culture stem cells, track defects in mice, and their dispensability. Key processes in hair follicle development highlighted include cadherin switching regulated by the Wnt and BMP signaling pathways and transcription factors like Lef1 and beta-catenin. Downregulation of E-cadherin, driven by these signals, is implicated in promoting polarity changes required for budding morphogenesis.
This study investigated the antioxidant effects of nebivolol in protecting against testicular damage caused by torsion-detorsion injury in rats. Forty rats were divided into four groups: a control group, a torsion group, a torsion/detorsion group, and a torsion/detorsion+nebivolol group. Biochemical assays and histopathological examination found that torsion-detorsion injury increased oxidative stress markers and apoptosis in testicular tissue, while administration of nebivolol before detorsion decreased oxidative stress and apoptosis. The study suggests that nebivolol has a protective effect against ischemia-reperfusion injury in the testes caused by torsion-
Objective: To identify interstitial cells of Cajal (ICC) in the common bile duct of Kunming mice.
Study Design: Common bile ducts obtained from the Kunming mice were prepared for immunohistochemical investigations using the c-kit antibody. Immunoelectron microscopy was used to detect the expression of c-kit in the ICC of the common bile duct. Transmission electron microscopy showed ultrastructure of ICC in the murine bile duct. Reverse transcription–polymerase chain reaction (RT-PCR) and western blot were used to confirm the expression of mRNA specific for the c-kit gene and production of c-kit protein in the Kunming mice common bile duct.
Results: Immunohistochemistry revealed that ICC in the murine common bile duct are c-kit positive and the ICC are located in the tela submucosa and the tunica muscularis of the murine common bile duct and do not connect with each other. Immunoelectron microscopy confirmed the expression of Kit by ICC in the murine common bile duct. Transmission electron microscopy showed that ICC in the murine common bile duct have long processes, abundant mitochondria, plenty of smooth endoplasmic reticulum (sER), a lot of lysosomes, and dense bodies. The caveolae of ICC are distinctive. At the same time, RT-PCR indicated that the Kunming mice common bile duct expressed mRNA specific for the c-kit gene, and western blot analysis showed the evidence of production of c-kit protein in the Kunming mice common bile duct.
Conclusion: ICC are found in the Kunming mice common bile duct, which is likely to lead to the development of motility study of the common bile duct.
Keywords: common bile duct; electron microscopy; immuno-electron microscopy; interstitial cells of Cajal; intestines; smooth muscle; tyrosine kinase receptor (c-kit)
Ldb 145 Geni Mutanti_2014-11-19 Jamora - dalla ricerca al prodotto 5laboratoridalbasso
1) Fibulin-5 expression is elevated in the dermis of Snail transgenic skin, a model of tissue fibrosis. Deletion of Fibulin-5 decreases tissue stiffness and the amount of elastic fibers but not collagen fibers.
2) Depletion of Fibulin-5 decreases inflammation in the fibrotic skin as measured by reduced macrophage and T cell infiltration.
3) Fibulin-5 promotes the production of inflammatory chemokines from dermal fibroblasts and accentuates their activation both in vivo and in response to substrate stiffness in vitro.
Ld b 145 geni mutanti_2014-11-18 jamora - ricerca scientifica 3laboratoridalbasso
This document discusses the role of the transcription factor Snail in wound healing and fibrosis. It finds that overexpression of Snail in the epidermis of mice induces dermal fibrosis through increased secretion of spondin-2/mindin by keratinocytes. Spondin-2 then activates dermal fibroblasts by stimulating the NF-kB pathway, leading to increased expression of genes associated with myofibroblast differentiation and extracellular matrix production. This establishes a paracrine signaling mechanism from epidermal Snail to induce dermal fibrosis through spondin-2.
Spermatotoxic impact of bonny light crude oil (BLCO) ingestion on adult male ...lukeman Joseph Ade shittu
Increasing concern has been expressed about the possible declining trend in the sperm quality and sperm count of man as a result of exposure to environmental estrogenic agents in the past few years now. There is a general paucity of knowledge of BLCO ingestion on the reproductive effect. Hence, we aim to evaluate the impact of sub-lethal dose of BLCO ingestion on semen parameters of adult male mice. Initial acute toxicity study was carried out to determine the lethal dose of BLCO, which was calculated to be 37.4 mg/Kg body wt. A sub-lethal dose of 20 mg/Kg bwt /day of BLCO were then given to 8 male mice in the experimental group. While, the control group of 7 animals received equal volume of 0.9% normal saline via oral garvage for 2 weeks. Data were analysed using SPSS 12 statistical software with P < 0.05 considered statistically significant. There was a significant (P < 0.05) weight gain in the treated group with a significant (P < 0.05) reduction in sperm motility in the treated compared with control. The sperm density of treated and control were 14.5 x 106 /ml and 20.5 x 106 /ml respectively. However, there were also no significant difference in the relative testicular weight and sperm density of treated from that of the control respectively. Thus, it was concluded that BLCO ingestion is spermatotoxic in the adult male Swiss mice
This study investigated the effects of gallic acid on testicular injury caused by ischemia-reperfusion in a rat testicular torsion model. Forty rats were divided into four groups: a control group, a torsion group, a torsion/detorsion group, and a torsion/detorsion plus gallic acid group. Biochemical markers and immunohistochemical staining for caspase-3 and TNF-α were analyzed. The results showed that gallic acid treatment decreased oxidative stress markers, reduced apoptosis and inflammation, and helped protect testicular tissue compared to the torsion/detorsion group without treatment. The study suggests that gallic acid may be a potential therapeutic agent for testicular ischemia-reperfusion injury.
Zebrafish are a popular model organism used in research due to their low cost, short lifecycle, optical transparency during development, and genetic similarity to humans. They can be used to study various types of toxicity including developmental toxicity, cardiotoxicity, neurotoxicity, and organ-specific toxicity. While zebrafish have limitations compared to mammalian models, assays using zebrafish embryos and larvae can provide early stage screening of compounds to evaluate teratogenicity and predict toxicity in mammals.
1) The document describes an experiment that examined the effects of tamoxifen (TAM) and other hormones on male and female mouse mammary gland development. Mice were treated with TAM, estrogen, or combinations of hormones for two weeks.
2) The mammary glands were then dissected and analyzed as whole mounts. Variables like gland elongation, number of end buds, and branch points were measured and compared between treatment groups and sexes.
3) As expected, TAM had little effect on male mouse mammary glands compared to controls, while female glands were more developed. Estrogen and combinations of hormones generally increased variables for both sexes, but female glands remained more developed than males.
34.Isolation and purification of juvenile hormone of Bombyx moriAnnadurai B
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Sodium Thiosulfate (Hydrogen Sulfide Donor): Ameliorates the Pituitary-testic...BRNSSPublicationHubI
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The five articles summarize and compare different tests for detecting bacterial endotoxins, including the rabbit pyrogen test and Limulus Amebocyte Lysate (LAL) test. The rabbit pyrogen test involves injecting a sample into rabbits and monitoring their temperatures, but it has limitations for certain vaccines. The LAL test detects endotoxins in vitro and is more sensitive than the rabbit test. Newer tests involving human cell lines are being developed as alternatives to better predict human responses to pyrogens.
The document describes a lab experiment that tests how the addition of a pGLO plasmid affects the growth and characteristics of E. coli bacteria. The experiment involves transforming E. coli bacteria with the pGLO plasmid by adding it to a solution containing the bacteria. One solution receives the pGLO plasmid (+pGLO) while the other does not (-pGLO). The bacteria are then observed under UV light and incubated under various conditions to analyze effects on growth and gene expression.
Anti-Fertility Effects of Nicotiana tabacum Leaf Smoke on the Spermatogenic Cells of the Adult Male Wistar Rats Testis
http://dx.doi.org/10.21276/SSR-IIJLS.2020.6.3.7
1) The study evaluated the antifertility effects of an alcoholic extract of Areca catechu in male albino rats.
2) Rats treated with 300mg/kg and 600mg/kg of the extract showed 50% and 100% reduction in fertility respectively.
3) Histological examination of the testes showed effects like reduction in sperm production and changes in testicular structure at higher doses.
Enterocin 55 produced by non rabbit-derived strain Enterococcus faecium EF55 ...Agriculture Journal IJOEAR
This document summarizes a study that investigated the effects of enterocin 55 (Ent55), produced by Enterococcus faecium EF55, on the microbiota and health parameters of broiler rabbits. Ent55 was administered to an experimental group of rabbits for 3 weeks. Microbial analysis found that Ent55 reduced counts of coagulase-negative staphylococci, Pseudomonas species, and coliforms in fecal and intestinal samples. Ent55 also increased phagocytic activity and reduced Eimeria oocyst counts, while not negatively impacting growth performance or biochemical parameters. The results indicate that Ent55 produced by a non-native strain can provide protective and beneficial effects in broiler rabbits.
Nicotine exposure during fetal development and lactation alters the intra-ovarian insulin-like growth factor (IGF) system in rats. Specifically:
1) Rats exposed to nicotine prenatally and through weaning had reduced IGF-I expression and IGFR-II mRNA expression in the ovary as adults.
2) There was no effect of nicotine exposure on mRNA expression levels of other IGF system components like IGF-II, IGFR-I, or IGF binding proteins.
3) The results suggest that decreased fertility and increased follicular atresia in nicotine-exposed rats may be partly due to disruption of IGF regulation in the ovary through reduction
This study investigated the protective effects of losartan, an AT1 receptor blocker, on testicular injury caused by ischemia/reperfusion in a rat testicular torsion model. Forty rats were divided into four groups: a control group, a torsion group, a torsion/detorsion group, and a torsion/detorsion plus losartan group. Biochemical assays and histopathological analysis showed that losartan prevented oxidative damage and reduced apoptosis in germ cells compared to the torsion/detorsion group, suggesting losartan has a protective role against ischemia/reperfusion injury in rat testes.
Age Related Histomorphological and Transmission Electron Microscopic Studies ...iosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
The Importance Of Animal Uses In AnimalsJessica Lopez
This document discusses using bromophenols as potential therapeutics for treating type 2 diabetes mellitus (T2DM). Bromophenols have been identified as inhibitors of the enzyme tyrosine phosphatase 1B (PTP1B), which is involved in insulin signaling. A series of new bromophenol analogs will be synthesized and tested for PTP1B inhibitory activity using in vitro enzyme assays to elucidate their mechanism of action. Bromophenols occur naturally in marine organisms and have attracted interest as anti-diabetic agents due to their suspected PTP1B inhibitory activity. However, quinone species formed from bromophenols could be toxic, so further study is needed to
Microanatomical and Hormonal Studies of the Effects of Aqueous Cannabis sativa Leaf Extract on the Testis of Adult Wistar Rats
http://dx.doi.org/10.21276/SSR-IIJLS.2020.6.4.1
39.Testicular enzymes in silkworms eposed to high temperature and RM values o...Annadurai B
This document reports on a study that investigated the effects of high temperature and mating on testicular enzymes and band patterns in silkworm moths. The study found that some enzymes like succinate and glutamate dehydrogenases increased sperm production, while malate dehydrogenase reduced it. Esterase enzymes were important for copulation. Electrophoresis revealed different band patterns for virgin, mated, and egg-laying moths. Temperature and humidity in January were found to be most favorable for sperm and egg production.
IBMX part I -rabbit reproduction Theriogenology 2010 (1)ELENI FOTOPOULOU
This document summarizes a study that examined the effects of 3-isobutyl-1-methyl-xanthine (IBMX), an inhibitor of cyclic adenosine monophosphate and cyclic guanosine monophosphate phosphodiesterases, on rabbit reproduction. The study found that IBMX enhanced the stimulatory effects of gonadotropins on ovulation, embryo development, and fertility. IBMX increased the number of ovulations, embryos, and pregnancy and birth rates. Younger rabbits were more sensitive to IBMX than older rabbits. The results suggest that cyclic nucleotides play an important role in controlling rabbit reproduction and that IBMX can stimulate reproduction and fertility.
Neuroprotective Roles of Oleic Acid: An Antioxidant Status and Cerebellar Cha...semualkaira
Previous studies have linked Alzheimer’s disease (AD) with cerebellar dysfunction. However, these studies
lack substantial cerebellar characterization and features during AD
progression.
Similar to The effect of varied doses of nicotine on some morphometric (20)
Abnormalities of hormones and inflammatory cytokines in women affected with p...Alexander Decker
Women with polycystic ovary syndrome (PCOS) have elevated levels of hormones like luteinizing hormone and testosterone, as well as higher levels of insulin and insulin resistance compared to healthy women. They also have increased levels of inflammatory markers like C-reactive protein, interleukin-6, and leptin. This study found these abnormalities in the hormones and inflammatory cytokines of women with PCOS ages 23-40, indicating that hormone imbalances associated with insulin resistance and elevated inflammatory markers may worsen infertility in women with PCOS.
A usability evaluation framework for b2 c e commerce websitesAlexander Decker
This document presents a framework for evaluating the usability of B2C e-commerce websites. It involves user testing methods like usability testing and interviews to identify usability problems in areas like navigation, design, purchasing processes, and customer service. The framework specifies goals for the evaluation, determines which website aspects to evaluate, and identifies target users. It then describes collecting data through user testing and analyzing the results to identify usability problems and suggest improvements.
A universal model for managing the marketing executives in nigerian banksAlexander Decker
This document discusses a study that aimed to synthesize motivation theories into a universal model for managing marketing executives in Nigerian banks. The study was guided by Maslow and McGregor's theories. A sample of 303 marketing executives was used. The results showed that managers will be most effective at motivating marketing executives if they consider individual needs and create challenging but attainable goals. The emerged model suggests managers should provide job satisfaction by tailoring assignments to abilities and monitoring performance with feedback. This addresses confusion faced by Nigerian bank managers in determining effective motivation strategies.
A unique common fixed point theorems in generalized dAlexander Decker
This document presents definitions and properties related to generalized D*-metric spaces and establishes some common fixed point theorems for contractive type mappings in these spaces. It begins by introducing D*-metric spaces and generalized D*-metric spaces, defines concepts like convergence and Cauchy sequences. It presents lemmas showing the uniqueness of limits in these spaces and the equivalence of different definitions of convergence. The goal of the paper is then stated as obtaining a unique common fixed point theorem for generalized D*-metric spaces.
A trends of salmonella and antibiotic resistanceAlexander Decker
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A transformational generative approach towards understanding al-istifhamAlexander Decker
This document discusses a transformational-generative approach to understanding Al-Istifham, which refers to interrogative sentences in Arabic. It begins with an introduction to the origin and development of Arabic grammar. The paper then explains the theoretical framework of transformational-generative grammar that is used. Basic linguistic concepts and terms related to Arabic grammar are defined. The document analyzes how interrogative sentences in Arabic can be derived and transformed via tools from transformational-generative grammar, categorizing Al-Istifham into linguistic and literary questions.
A time series analysis of the determinants of savings in namibiaAlexander Decker
This document summarizes a study on the determinants of savings in Namibia from 1991 to 2012. It reviews previous literature on savings determinants in developing countries. The study uses time series analysis including unit root tests, cointegration, and error correction models to analyze the relationship between savings and variables like income, inflation, population growth, deposit rates, and financial deepening in Namibia. The results found inflation and income have a positive impact on savings, while population growth negatively impacts savings. Deposit rates and financial deepening were found to have no significant impact. The study reinforces previous work and emphasizes the importance of improving income levels to achieve higher savings rates in Namibia.
A therapy for physical and mental fitness of school childrenAlexander Decker
This document summarizes a study on the importance of exercise in maintaining physical and mental fitness for school children. It discusses how physical and mental fitness are developed through participation in regular physical exercises and cannot be achieved solely through classroom learning. The document outlines different types and components of fitness and argues that developing fitness should be a key objective of education systems. It recommends that schools ensure pupils engage in graded physical activities and exercises to support their overall development.
A theory of efficiency for managing the marketing executives in nigerian banksAlexander Decker
This document summarizes a study examining efficiency in managing marketing executives in Nigerian banks. The study was examined through the lenses of Kaizen theory (continuous improvement) and efficiency theory. A survey of 303 marketing executives from Nigerian banks found that management plays a key role in identifying and implementing efficiency improvements. The document recommends adopting a "3H grand strategy" to improve the heads, hearts, and hands of management and marketing executives by enhancing their knowledge, attitudes, and tools.
This document discusses evaluating the link budget for effective 900MHz GSM communication. It describes the basic parameters needed for a high-level link budget calculation, including transmitter power, antenna gains, path loss, and propagation models. Common propagation models for 900MHz that are described include Okumura model for urban areas and Hata model for urban, suburban, and open areas. Rain attenuation is also incorporated using the updated ITU model to improve communication during rainfall.
A synthetic review of contraceptive supplies in punjabAlexander Decker
This document discusses contraceptive use in Punjab, Pakistan. It begins by providing background on the benefits of family planning and contraceptive use for maternal and child health. It then analyzes contraceptive commodity data from Punjab, finding that use is still low despite efforts to improve access. The document concludes by emphasizing the need for strategies to bridge gaps and meet the unmet need for effective and affordable contraceptive methods and supplies in Punjab in order to improve health outcomes.
A synthesis of taylor’s and fayol’s management approaches for managing market...Alexander Decker
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A survey paper on sequence pattern mining with incrementalAlexander Decker
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A survey on live virtual machine migrations and its techniquesAlexander Decker
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This document surveys trust architectures that leverage provenance in wireless sensor networks. It begins with background on provenance, which refers to the documented history or derivation of data. Provenance can be used to assess trust by providing metadata about how data was processed. The document then discusses challenges for using provenance to establish trust in wireless sensor networks, which have constraints on energy and computation. Finally, it provides background on trust, which is the subjective probability that a node will behave dependably. Trust architectures need to be lightweight to account for the constraints of wireless sensor networks.
This document discusses private equity investments in Kenya. It provides background on private equity and discusses trends in various regions. The objectives of the study discussed are to establish the extent of private equity adoption in Kenya, identify common forms of private equity utilized, and determine typical exit strategies. Private equity can involve venture capital, leveraged buyouts, or mezzanine financing. Exits allow recycling of capital into new opportunities. The document provides context on private equity globally and in developing markets like Africa to frame the goals of the study.
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Integrating Ayurveda into Parkinson’s Management: A Holistic ApproachAyurveda ForAll
Explore the benefits of combining Ayurveda with conventional Parkinson's treatments. Learn how a holistic approach can manage symptoms, enhance well-being, and balance body energies. Discover the steps to safely integrate Ayurvedic practices into your Parkinson’s care plan, including expert guidance on diet, herbal remedies, and lifestyle modifications.
- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
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NVBDCP.pptx Nation vector borne disease control programSapna Thakur
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Histololgy of Female Reproductive System.pptxAyeshaZaid1
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Osteoporosis - Definition , Evaluation and Management .pdfJim Jacob Roy
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Cell Therapy Expansion and Challenges in Autoimmune DiseaseHealth Advances
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Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
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Rasamanikya is a excellent preparation in the field of Rasashastra, it is used in various Kushtha Roga, Shwasa, Vicharchika, Bhagandara, Vatarakta, and Phiranga Roga. In this article Preparation& Comparative analytical profile for both Formulationon i.e Rasamanikya prepared by Kushmanda swarasa & Churnodhaka Shodita Haratala. The study aims to provide insights into the comparative efficacy and analytical aspects of these formulations for enhanced therapeutic outcomes.
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The effect of varied doses of nicotine on some morphometric
1. Journal of Natural Sciences Research www.iiste.org
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.7, 2013
187
The Effect of Varied Doses of Nicotine on Some Morphometric
Parameters of the Testis in Albino Wistar Rats
Nanyak Z. Galam*1
, Ibraheem M. Gambo 1
,Zingkur Z. Galam2
, Gomerep S. Simji3
, Ayaka Louis O1
, Simi
Dami1
, Gotom S. Sale5
,Vhriterhire A.Raymond4
, U.G Egesie1
1. Department of Human physiology, University of Jos.
2. Department of Surgery, Jos University teaching hospital.
3. Department of Internal medicine, University of Jos
4. Department of Pathology, University of Jos.
5. Department of Anatomy University of Jos.
*galam_n@yahoo.com – Correspponding Author.
Abstract
Background: This study is aimed at determining the effect of nicotine on male fertility by evaluating some
morphometric parameters of male Wistar rat such as testicular weight, seminiferous tubule diameter, height of
epithelium, sertoli-germ cell ratio and Johnsen’s score in order to assess the spermatogic index.
Methods: 20 adult male rats were randomly divided into four groups, the test groups were administered with
0.2mg/100g, 0.4/100g and 0.6/100g body weight of nicotine base daily for 30 days using a polythene catheter
orally , while the control were administered with 2mls 0.9%physiological saline. Histological slides of the testis
were made and data obtained from the slides.
Results: nicotine caused a significant reduction (P < 0.05) and (P< 0.01) in the mean values of the morphometric
parameters of the test group compared with control. The Johnsen’s Score results were 5.4 ± 0.51*, 5.2 ± 0.37*
for groups 2 and 3 respectively. For seminiferous tubule diameter the significant values were 208.0±4.22**,
184.5±11.31**in groups 2 and 3, whilst the height of epithelium showed a significant reduction of 153.9±
13.58*, 101.6 ± 1.35**, 76.6 ± 6.65** for groups 1, 2 and 3 respectively. Sertoli-germ cell ratio showed a
significant reduction in groups 2 and 3 with values of 3.6 ± 0.77 ** , 1.5 ± 0.16**for groups 2, and 3
respectively, testicular weight showed significant reduction of 1.2±0.05**,1.1±0.05** for group 2 and 3.The
results stated are only those that showed a significant reduction at 95% confidence level when compared with the
control.
Conclusion: It was concluded that nicotine exerted an adverse effects on the spermatogenic index with
concomitant reduction in reproductive potentials of the male rat. Nicotine and nicotine- based products should
therefore be taken with caution in cases of infertility in man and animal.
Key words: Morphometric parameters ,Johnsen’s Score,Spermatogenic index , fertility
Infertility can be defined as the failure to conceive despite having regular unprotected sexual intercourse for a
period of twelve months. It is said that approximately one in ten people suffer from this disability with 50% of
the causes being attributed to the male factor (De Kretser and Baker 1999) though many factors associated with
female infertility have been mentioned with subsequent advances in the management of infertility amongst them,
not much has been done in respect to managing male infertility. Some factors have however been listed as
possible causes of male infertility prominent among them is a previous history of sexually transmitted diseases.
Other factors like tobacco smoking have been mentioned though more with female infertility but have also been
implicated in male infertility.
Nicotine is an alkaloid found in the nightshade family of plants (Solanaceae); biosynthesis takes place in the
roots and accumulation occurs in the leaves. It constitutes approximately 0.6–3.0% of the dry weight of tobacco
(NIH) and is present in the range of 2–7 µg/kg of various edible plants. It functions as an antiherbivore chemical;
therefore, nicotine was widely used as an insecticide in the past ( Rodgman et al, 2009) and nicotine analogue
such as imidacloprid are currently widely used. It being a central nervous system influencing drug interferes in
many endocrine activities related to reproduction. It inhibits the release of FSH and LH from the pituitary (Blake
et al., 1972; Blake, 1974, 1978). Nicotine administration or exposure to cigarette smoke inhalation to rats and
hamsters results in testicular degeneration (Viczian, 1968; Tsilikov, 1969). A study done by Gambo et al (2013)
showed dose dependent testicular degeneration following the administration of varied doses of an aqueous
extract of tobacco.
2. Journal of Natural Sciences Research www.iiste.org
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.7, 2013
188
Though there are studies such as this, the incidence of tobacco smoking is said to be on the rise as of 2002, about
twenty percent of young teens (13-15) smoke worldwide. 80,000-100,000 children begin smoking every day.
Half of those who begin smoking in adolescent years are projected to go on to smoke for 15 to 20 years (WHO,
2002). In the developing world tobacco consumption is rising by 3.4% annually (WHO, 2002).
The rising incidence in tobacco smoking particularly in the developing world as well as the little knowledge of
the effect of tobacco on male fertility in comparison to the females necessitates studies such as these to
determine the effect nicotine on spermatogenesis, whilst making an attempt to count and score the germ cells
using the male albino wistar rat as a model.
MATERIALS AND METHODS
Animals
20 male Wistar rats weighing 220 - 280 g were used in the study. All animals were kept in the animal house of
the University of Jos. They were maintained at room temperature and 12 hours light/dark cycle. All the
experimental procedures were done following the experimental guidelines of Institutional Animal Ethics
Committee (IAEC).
Chemicals
Testosterone, Oestrogen, FSH, LH, Progesterone, EIA(Enzyme-linked Immunosorbent Assay) kit ways
purchased from Monobind Inc. Lake forest U.S.A. Nicotine hydrogen tartrate salt (C10H14N22C4H6O6)
purchased from sigma Aldrich catalog number N5260-25G.
Experimental protocol
Four groups with 5 rats selected randomly in each group were formed. Groups 1-3 were the test groups, whilst
group 4 was used as the control, each rat in group 1 was treated with 0.2 mg/100g body weight /p.o of nicotine
daily for 30 days, groups 2 and 3 were given 0.4mg/ 100g and 0.6mg/ 100g body weight of nicotine per os daily
for 30 days respectively. While group 4 (control group) was given 2ml of 0.9% physiological saline solution for
the same period of exposure.
Sample collection
The rats were anaesthetized using ether and afterward sacrificed by cervical dislocation and blood sample
collected by cardiac punc-ture. Orchidectomy was performed by open castration method. A midline or pre-
scrotal incision was made and the testicles were milked out of the incision site and weighed with the aid of
OHAUS electric weighing balance.
The testicles were exposed by incising the tunica vaginalis.
Histological procedures
After the extraction of the testis from the animal’s body, the organ was promptly and adequately treated with
10% formal saline (fixation) in order to preserve its structure and molecular composition. After fixation, the
piece of organ was dehydrated by bathing it successfully in graded mixture of ethanol and water (70 - 100%).
The ethanol was then replaced with a solvent miscible with the embedding medium (xylene). As the tissues were
infiltrated with xylene, it became transparent (clearing). Once the tissue has been impregnated by xylene it was
placed in melted paraffin in an oven maintained at 58 - 60°C (embedding). The heat caused the solvent to
evaporate and the spaces within the tissues become filled with paraffin. The tissue together with its impregnating
paraffin hardens after been taken out of the oven this was done with the aid of the Leica TP120 automatic tissue
processor. The hard block containing the tissue was then taken to the microtome and sectioned by the microtome
steel. The sections were then floated on water and transferred to a glass slide and stained with heamatoxylin and
eosin stains with the aid of the leica auto stainer XL . The slides were then viewed under light microscope with
varying magnification. Spermatogenesis was assessed by a method which depended upon scoring ‘cross
sectional’ profiles of seminiferous tubules according to Johnsen’scriteria. (Johnsen S.G1970) The criteria use
scores from 1 to10 and are as follows: 10. Complete spermatogenesis with many spermatozoa. 9. Many
spermatozoa present but germinal epithelium disorganized with marked sloughing or obliteration of lumen. 8.
Only a few (<5) spermatozoa. 7. No spermatozoa but many spermatids. 6. No spermatozoa and only a few
spermatids (<5). 5. No spermatozoa or spermatids but many spermatocytes. 4. No spermatozoa or spermatids
and only a few spermatocytes(<5). 3. Spermatogonia are the only germ cells present. 2. No germ cells but sertoli
cells arepresent. 1. No cells in tubular section.
Diameter of seminiferous tubules was measured using Leica 1000 DM microscope and X10 objective lens with
ocular micrometer whilst the measurement for the height of epithelium was done at X40 magnification. The
sertoli/germ cell ratio was also determined, using stage 7 spermatids as the chosen line of germ cells on
photomicrographs of the histological slides. The pathologist involved in the count was blind to the grouping and
dosage of nicotine given to the rats.
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Statistical analysis
Statistical analysis was done using graphpad instat3 tool to conduct one way analysis of variance (ANOVA). The
graphpad prism 6 was used to generate a bar chart of the grouped data.
Results
Are presented as the mean ± standard error of mean. The results showed a significant dose dependent decrease at
all dose levels for the height of the epithelium, whilst showing a significant decrease in seminiferous tubule
diameter, Johnsen’s score and sertoli – germ cell ratio at the doses of 0.4mg/100g and 0.6mg/100g of nicotine
base only.
Figure 1: bar chart showing morphometric parameters measured.
Discussion : The sertoli- germ cell ratio, Johnsen’s score, testicular weight, height of the germinal epithelium
and seminiferous tubule diameter were used in this study to evaluate the effect of administration of varied doses
of nicotine over a period of thirty days on spermatogenesis, using the Wistar rat as animal model. The sertoli to
elongated germ cell ratio as well as the Johnsen’s score was used to evaluate the spermatogenic index, a decrease
in the sertoli to elongated spermatid ratio or Johnsen score indicates a fall in spermatogenesis (Aruna et al 2009).
The results showed a significant dose dependent decrease at all dose levels for the height of the epithelium,
whilst showing a significant decrease in seminiferous tubule diameter, Johnsen’s score and sertoli – germ cell
ratio at the doses of 0.4mg/100g and 0.6mg/100g of nicotine base only, corroborating the findings of Egesie et al
(2013) at the preliminary stage of this study, which reported the histological slides as having a dose dependent
decrease on spermatogenic cells and distortion of the cyto-architecture of the seminiferous tubule of the testis.
The decrease spermatogenic index is also in tandem with the findings of decrease sperm count in the study.
The finding of decreased sertoli to elongated germ cell ratio corroborates studies done by Rajpurkar (2000)
where they observed a decrease in the ratio as well as a decrease in seminiferous tubule diameter in rats exposed
to cigarettes smoke, which was also observed in this study, they how ever did not observe any significant
difference in the height of epithelium of the rats exposed to cigarettes smokes as opposed to this study which
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observed a significant difference in the height of the epithelium in the seminiferous tubules of tubules of the rats
given high doses of nicotine base. The observed difference in the findings may be as a result of the difference in
the modes of administration as well as the probable higher dose given as direct nicotine base in this study. The
observed decrease in the morphometric parameters measured may be as a result of the inhibition of the release of
gonadotrophins, follicle stimulating hormone and luteinizing from the anterior pituitary (Blake et al., 1972;
Blake, 1974, 1978). These hormones are necessary for proper development of the gonads as well as the process
of spermatogenesis.
Conclusion
Thus it can be stated from this study that the intake of nicotine especially at doses greater than 2mg/100g for a
period of 30 days in male albino wistar rats adversely altered the morphometric parameters measured with a
concomitant decrease in spermatogenic index, thus nicotine and nicotine – based products should be taken with
caution particularly in cases of infertility.
References
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