This document discusses Leishmania parasites' ability to reversibly inhibit muscle contractions in insects. It finds that:
1) L. major promastigotes and culture medium exhibit greatest myoinhibitory activity in late log and early stationary growth phases, coinciding with when infective forms develop in sand flies.
2) L. major lysate inhibits contractions of muscle preparations from cockroaches, bugs, and flies, indicating a conserved inhibitory mechanism across insect orders.
3) Other Leishmania species and gut-dwelling kinetoplastids also exhibit myoinhibitory activity in sand flies.
Inhibiting host gut contractions protects Leishmania from expulsion, allowing development and transmission.
1) L. major proteins (LMP) completely stopped spontaneous contractions of cultured rat cardiomyocytes, which resumed contracting after washing with saline.
2) LMP significantly decreased force of contractions in guinea pig ileum strips stimulated with nicotine in a dose-dependent manner, but had no effect on strips stimulated with acetylcholine. Contractions resumed after washing.
3) LMP also reduced contractions of guinea pig uterine strips stimulated with oxytocin, and of electrically stimulated guinea pig ileum strips, in a dose-dependent manner. Contractions resumed after washing in all cases.
1) A paralytic factor was isolated from crude lysates of Leishmania major promastigotes that caused reversible, dose-dependent inhibition of spontaneous contractions in isolated Phlebotomus papatasi hindguts.
2) Application of the L. major lysate to P. papatasi guts resulted in a 50-60% increase in midgut and hindgut volume after 30 minutes.
3) The L. major paralytic factor was purified over 100-fold and yielded a 12 kDa hydrophobic peptide. This peptide allows L. major to persist in the expanded, relaxed sand fly gut and facilitates transmission to vertebrate hosts.
This document summarizes a study that examined the bioconcentration and acute toxicity of the organophosphate pesticide methyl parathion in three species of Brazilian freshwater fish: pacu, piavussu, and curimbatá. Fish were exposed to 5 ppm of methyl parathion for various time periods. Tissue samples were taken to analyze pesticide concentration and cholinesterase activity in serum, liver, brain, heart, and muscle. Results showed similar bioconcentration of methyl parathion across fish species, with the highest concentrations found in brain tissue. Exposure to 5 ppm methyl parathion for 3-5 hours killed all curimbatá fish, associated with over 90% inhibition of
This document summarizes a research article that evaluated the immunomodulatory potential of crude protein extract from taro (Colocasia esculenta) corms on hematopoietic cells in mice. The crude taro extract stimulated in vitro proliferation of splenocytes from two mouse strains. When administered intraperitoneally to mice, the extract induced splenomegaly and proliferation of total spleen and bone marrow cells. It also promoted in vivo proliferation of B220+ splenocytes and affected levels of mature and immature B cells in bone marrow. The extract represents a source of immunostimulatory proteins with potential applications as food or pharmaceutical additives.
This study investigated the substrate specificity of the CYP1A enzyme from Pterygoplichthys sp., a species of catfish. The CYP1A gene from Pterygoplichthys sp. was expressed in yeast cells. The catalytic activity of the expressed enzyme was then tested against 15 potential substrates. Results showed that the enzyme had a much higher activity for coumarin derivatives than resorufin derivatives, unlike most other vertebrate CYP1As. The enzyme was also able to metabolize some flavones and ethoxyfluorescein but not resveratrol. These findings suggest the Pterygoplichthys sp. CYP1A has a divergent substrate
This document summarizes a research article that evaluated the safety risks of coagulase-negative staphylococci (CNS) present in the microbiota of commercial and artisanal salami in Brazil. Nineteen CNS strains were isolated from salami samples, with different species identified between commercial and artisanal salami. The strains were found to harbor multiple genes for enterotoxins and toxins and showed antimicrobial resistance. Real-time PCR and ELISA confirmed the isolated strains could express enterotoxins in vitro, posing risks for food poisoning. The study characterized the CNS species in Brazilian salami and their potential safety risks in terms of enterotoxin production and antimicrobial resistance.
The study screened extracts from 21 species of Clusiaceae native to Mexico for anti-HIV activity. Five species showed significant inhibition (≥70%) of HIV-1 reverse transcriptase. Of these, extracts from Calophyllum brasiliense (hexane) and Clusia quadrangula inhibited the enzyme and were non-toxic to human cells. The C. brasiliense extract was the most potent, inhibiting viral replication with an ED50 of 37.1 μg/ml, while the C. quadrangula extract was less active. The results support Clusiaceae as a source of potential anti-HIV compounds.
When yeast cells are exposed to anoxia (no oxygen) on a non-fermentable carbon source, they enter a state of suspended animation where all observable life processes reversibly halt until oxygen is restored. Transcriptional profiling revealed differences in gene expression between yeast exposed to carbon monoxide (CO) gas versus nitrogen (N2) gas. CO can mimic oxygen binding and led to derepression of aerobic metabolism genes compared to N2 exposure. Mutants lacking components of the mitochondrial retrograde signaling pathway recovered normally from CO but not N2 exposure, indicating its importance in the cellular response to different anoxic conditions. The study establishes yeast as a model for investigating suspended animation and oxygen-regulated gene expression.
1) L. major proteins (LMP) completely stopped spontaneous contractions of cultured rat cardiomyocytes, which resumed contracting after washing with saline.
2) LMP significantly decreased force of contractions in guinea pig ileum strips stimulated with nicotine in a dose-dependent manner, but had no effect on strips stimulated with acetylcholine. Contractions resumed after washing.
3) LMP also reduced contractions of guinea pig uterine strips stimulated with oxytocin, and of electrically stimulated guinea pig ileum strips, in a dose-dependent manner. Contractions resumed after washing in all cases.
1) A paralytic factor was isolated from crude lysates of Leishmania major promastigotes that caused reversible, dose-dependent inhibition of spontaneous contractions in isolated Phlebotomus papatasi hindguts.
2) Application of the L. major lysate to P. papatasi guts resulted in a 50-60% increase in midgut and hindgut volume after 30 minutes.
3) The L. major paralytic factor was purified over 100-fold and yielded a 12 kDa hydrophobic peptide. This peptide allows L. major to persist in the expanded, relaxed sand fly gut and facilitates transmission to vertebrate hosts.
This document summarizes a study that examined the bioconcentration and acute toxicity of the organophosphate pesticide methyl parathion in three species of Brazilian freshwater fish: pacu, piavussu, and curimbatá. Fish were exposed to 5 ppm of methyl parathion for various time periods. Tissue samples were taken to analyze pesticide concentration and cholinesterase activity in serum, liver, brain, heart, and muscle. Results showed similar bioconcentration of methyl parathion across fish species, with the highest concentrations found in brain tissue. Exposure to 5 ppm methyl parathion for 3-5 hours killed all curimbatá fish, associated with over 90% inhibition of
This document summarizes a research article that evaluated the immunomodulatory potential of crude protein extract from taro (Colocasia esculenta) corms on hematopoietic cells in mice. The crude taro extract stimulated in vitro proliferation of splenocytes from two mouse strains. When administered intraperitoneally to mice, the extract induced splenomegaly and proliferation of total spleen and bone marrow cells. It also promoted in vivo proliferation of B220+ splenocytes and affected levels of mature and immature B cells in bone marrow. The extract represents a source of immunostimulatory proteins with potential applications as food or pharmaceutical additives.
This study investigated the substrate specificity of the CYP1A enzyme from Pterygoplichthys sp., a species of catfish. The CYP1A gene from Pterygoplichthys sp. was expressed in yeast cells. The catalytic activity of the expressed enzyme was then tested against 15 potential substrates. Results showed that the enzyme had a much higher activity for coumarin derivatives than resorufin derivatives, unlike most other vertebrate CYP1As. The enzyme was also able to metabolize some flavones and ethoxyfluorescein but not resveratrol. These findings suggest the Pterygoplichthys sp. CYP1A has a divergent substrate
This document summarizes a research article that evaluated the safety risks of coagulase-negative staphylococci (CNS) present in the microbiota of commercial and artisanal salami in Brazil. Nineteen CNS strains were isolated from salami samples, with different species identified between commercial and artisanal salami. The strains were found to harbor multiple genes for enterotoxins and toxins and showed antimicrobial resistance. Real-time PCR and ELISA confirmed the isolated strains could express enterotoxins in vitro, posing risks for food poisoning. The study characterized the CNS species in Brazilian salami and their potential safety risks in terms of enterotoxin production and antimicrobial resistance.
The study screened extracts from 21 species of Clusiaceae native to Mexico for anti-HIV activity. Five species showed significant inhibition (≥70%) of HIV-1 reverse transcriptase. Of these, extracts from Calophyllum brasiliense (hexane) and Clusia quadrangula inhibited the enzyme and were non-toxic to human cells. The C. brasiliense extract was the most potent, inhibiting viral replication with an ED50 of 37.1 μg/ml, while the C. quadrangula extract was less active. The results support Clusiaceae as a source of potential anti-HIV compounds.
When yeast cells are exposed to anoxia (no oxygen) on a non-fermentable carbon source, they enter a state of suspended animation where all observable life processes reversibly halt until oxygen is restored. Transcriptional profiling revealed differences in gene expression between yeast exposed to carbon monoxide (CO) gas versus nitrogen (N2) gas. CO can mimic oxygen binding and led to derepression of aerobic metabolism genes compared to N2 exposure. Mutants lacking components of the mitochondrial retrograde signaling pathway recovered normally from CO but not N2 exposure, indicating its importance in the cellular response to different anoxic conditions. The study establishes yeast as a model for investigating suspended animation and oxygen-regulated gene expression.
This study examined the effects of beta glucan as an immunostimulant on the Indian major carp, Labeo rohita, challenged with the bacterial pathogen Aeromonas hydrophila. The experimental groups included: a control group, a group treated with beta glucan, a group treated with beta glucan and challenged with A. hydrophila, and a group only challenged with A. hydrophila. Results showed that beta glucan treatment improved growth performance and boosted hematological and biochemical parameters related to immunity compared to the control and infection-only groups. Specifically, beta glucan increased total erythrocyte count, hemoglobin levels, and improved condition factor and specific growth rate. It also elevated total leukocyte count and different white
This document summarizes a study that isolated and characterized lactic acid bacteria from various environmental samples. 21 lactic acid bacteria isolates were obtained from milk, water, soil and plant samples. 10 were identified as Lactobacillus, 3 as Enterococcus, 2 as Staphylococcus, 5 as Lactococcus, and 1 as Leuconostoc based on biochemical and physiological tests. 6 of the isolates were found to harbor plasmids. Further characterization identified 3 isolates as Enterococcus faecium and 1 each as Weissella confusa, Pediococcus pentosaceus, and Staphylococcus epidermidis based on 16S rRNA gene sequencing. Some isolates showed inhibitory activity
This document summarizes laboratory experiments on Pseudo-nitzschia species isolated from Monterey Bay, California. The experiments tested the effects of nutrient stress and culturing methods on toxin production and photosynthetic performance. Results showed substantial variability between clones in growth rates and toxicity levels under identical conditions. Toxin levels increased under silicon limitation and decreased with higher growth rates in chemostat experiments. Variable fluorescence measurements indicated nutrient stress impaired photosystem II and negatively correlated with toxin accumulation.
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)
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.
1. The document presents the discovery of JSI-124, a selective inhibitor of the JAK/STAT3 pathway, which was identified from screening a library of compounds for inhibition of STAT3 phosphorylation in cancer cells.
2. JSI-124 was found to reduce phospho-STAT3 levels in multiple human cancer cell lines and block STAT3 DNA binding and gene transcription, inhibiting tumor growth.
3. JSI-124 selectively targeted STAT3 signaling over other pathways and showed potent antitumor activity against human tumor xenografts in mouse models, demonstrating its potential as an anticancer therapeutic targeting the STAT3 pathway.
This document summarizes several studies that investigated methods for quantifying and tracking monocyte recruitment into atherosclerotic plaques. Initial studies in pigs used electron microscopy to observe monocyte movement into plaques. Subsequent studies labeled monocytes with fluorescent tags or genetic markers and tracked their movement into plaques in animal models like pigs, mice and rabbits. These studies found that labeled monocytes were detected within plaque lesions and that antibodies blocking adhesion molecules reduced monocyte recruitment, demonstrating the methods effectively tracked and quantified monocyte infiltration into plaques.
1) Several studies have traced the recruitment of monocytes into atherosclerotic plaques using labeling techniques. Gerrity et al labeled monocytes with FITC and found them adhered to plaque sites.
2) Willerson et al labeled macrophages with fluorescent microspheres and found them adhering to plaques. Antibodies to ICAM-1 and integrin reduced recruitment.
3) Steinberg et al transfused leukocytes between genetically different rabbits to track recruitment to plaques using PCR, finding over 600 donor cells per million in early fatty streaks and over 3,800 in advanced plaques.
Integrin V can form heterodimers with several subunits to mediate cell-cell and cell-extracellular matrix interactions. During zebrafish gastrulation, V is expressed maternally and zygotically. Here, we used a morpholino-mediated V knockdown strategy to study V function. Although V morphants displayed vascular defects, they also exhibited left-right body asymmetry defects affecting multiple visceral organs. This was preceded by mislocalization of dorsal forerunner cells (DFCs) and malformation of the Kupffer’s vesicle (KV) laterality organ
Commercial Application of Anoectochilus formosanus: Immunomodulating ActivitiesCây thuốc Việt
Anoectochilus formosanus is an important ethnomedicinal plant of Taiwan. We investigated the effect of oral administration of A. formosanus effective fraction (AFEF) on the innate immune response in mice. Male BALB/c mice were treated orally for 2 weeks with 500, 1000 and 1500 mg/kg of AFEF. Primary peritoneal macrophage harvest from mice that administered with AFEF (500 –1500 mg/kg) was directed to activate phagocytosis. AFEF significantly increased interferon-production from lymph node cells by ConA stimulation for 48 hours in AFEF (1500 mg/kg) treated group. AFEF might be the active fraction in activation of innate immunity.
1) Transgenic fish models carrying bacteriophage λ and plasmid pUR288 vectors were developed to improve methods for assessing health risks from environmental mutagens and establish new animal models for studying in vivo mutagenesis.
2) The bacteriophage λ transgenic medaka model uses the cII and lacI genes as mutational targets, allowing detection of mutations through packaging of the phage vector and infection of E. coli. Spontaneous mutation frequencies in medaka were comparable to rodent models.
3) Exposure to chemical mutagens like ENU induced concentration-dependent, tissue-specific, and time-dependent increases in cII mutations, demonstrating the utility of the transgenic fish model for studying mut
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 document describes two experiments that examined the effects of dietary zinc source and coccidial vaccine exposure on zinc homeostasis and immune status in broiler chickens. The experiments found that coccidial challenge decreased intracellular zinc levels and phagocytic capacity in the jejunum, while increasing phagocytic capacity in the caecal tonsils. Coccidial challenge also increased the ratio of zinc import to export transporters. Dietary zinc source had little impact except on one zinc transporter. The results suggest that during coccidial challenge, intestinal cells attempt to compensate for the drop in intracellular zinc by upregulating zinc import transporters.
The experiment aimed to determine if inserting arginine-producing enzymes from Chlorella vulgaris into a mutated form of Chlamydomonas reinhardtii lacking the arg2 gene would allow the mutated algae to grow. Arginine-producing enzymes were extracted from C. vulgaris and added to growth medium containing the mutated C. reinhardtii. There was growth of the mutated C. reinhardtii with the inserted enzymes without arginine supplements, supporting the hypothesis that the enzymes would allow the algae to produce arginine and grow. Control groups of C. reinhardtii grown with and without supplemental arginine showed different growth patterns.
Widespread Natural Occurrence of Hydroxyurea in AnimalsMichelle Pyle
This research article reports the discovery of naturally occurring hydroxyurea, an antibiotic compound, in many animal species across taxonomic groups. The highest levels were found in the little skate shark, with concentrations in some tissues high enough to have antiviral and anticancer effects based on in vitro studies. Embryos of the little skate also showed increasing hydroxyurea levels with development, indicating the ability to synthesize it. While levels in most other species examined were lower, certain tissues in some invertebrates and vertebrates also had concentrations expected to have antiviral effects. The widespread natural presence of hydroxyurea suggests it may play a role in disease resistance as part of the innate immune system in combating infections.
6 magalhães et al 2008 a hyaluronidase from potamotrygon motoro (freshwaterpryloock
1) Researchers purified and characterized a hyaluronidase enzyme from the venom of the freshwater stingray Potamotrygon motoro.
2) A two-step purification process involving gel filtration chromatography and ion exchange chromatography resulted in a 366-fold purification with a 79 kDa hyaluronidase enzyme.
3) The purified hyaluronidase had a pH optimum of 4.2 and maximum activity at 40°C. Its activity was inhibited by certain metal ions and heparin.
This document summarizes a study that investigated how mechanical forces applied to integrin receptors control intracellular signaling in osteoblasts. The researchers found that cyclic forces applied to the beta-1 integrin subunit at 1 Hz were more effective at stimulating calcium responses in osteoblasts than continuous forces. Cyclic forces also induced increased tyrosine phosphorylation of cytoskeleton-anchored proteins and greater activation of focal adhesion kinase and mitogen-activated protein kinase compared to continuous forces. These responses depended on an intact cytoskeleton and the presence of intracellular calcium. Analysis of spatial calcium signals revealed they originated near the stressed receptors, indicating cells can sense local stress via integrins.
IOSRPHR(www.iosrphr.org) IOSR Journal of Pharmacyiosrphr_editor
This document summarizes research on developing polyclonal antibodies to identify isoflavones in different legume species. Key points:
- Researchers produced polyclonal antibodies in rabbits to detect isoflavones. ELISA tests using these antibodies could quickly screen plants for isoflavone content.
- Seed samples from 8 legume species were tested for isoflavones using the ELISA. All species showed higher isoflavone levels than soybean (the typical isoflavone source), indicating other legumes may be alternative isoflavone sources.
- Extraction methods including hydrothermal treatment at 50°C increased levels of active aglycone isoflavone forms compared to no treatment.
- The research aims to characterize legume
Biodiversity conservation has conventionally dealt with management aspects, but over the last decade the issue of governance, i.e. who decides and how, has gained prominence, resulting in crucial paradigm shifts in protected area and other conservation policies and practice.
The document discusses the key components of language construction:
1) Primitives are the smallest units of meaning that cannot be broken down further, such as morphemes in natural languages.
2) Means of combination are rules for building words from primitives and phrases/sentences from words.
3) Means of abstraction allow assigning a simple name to a complex entity, such as pronouns in English that take on meaning based on context.
Este documento explica los cuatro casos gramaticales en alemán (nominativo, dativo, acusativo y genitivo) y cómo se relacionan con las funciones gramaticales en español como sujeto, complemento indirecto, complemento directo y complemento del nombre. También incluye una tabla detallada que muestra cómo declinar sustantivos, adjetivos y pronombres posesivos en alemán según el caso, género, número y terminación.
This study examined the effects of beta glucan as an immunostimulant on the Indian major carp, Labeo rohita, challenged with the bacterial pathogen Aeromonas hydrophila. The experimental groups included: a control group, a group treated with beta glucan, a group treated with beta glucan and challenged with A. hydrophila, and a group only challenged with A. hydrophila. Results showed that beta glucan treatment improved growth performance and boosted hematological and biochemical parameters related to immunity compared to the control and infection-only groups. Specifically, beta glucan increased total erythrocyte count, hemoglobin levels, and improved condition factor and specific growth rate. It also elevated total leukocyte count and different white
This document summarizes a study that isolated and characterized lactic acid bacteria from various environmental samples. 21 lactic acid bacteria isolates were obtained from milk, water, soil and plant samples. 10 were identified as Lactobacillus, 3 as Enterococcus, 2 as Staphylococcus, 5 as Lactococcus, and 1 as Leuconostoc based on biochemical and physiological tests. 6 of the isolates were found to harbor plasmids. Further characterization identified 3 isolates as Enterococcus faecium and 1 each as Weissella confusa, Pediococcus pentosaceus, and Staphylococcus epidermidis based on 16S rRNA gene sequencing. Some isolates showed inhibitory activity
This document summarizes laboratory experiments on Pseudo-nitzschia species isolated from Monterey Bay, California. The experiments tested the effects of nutrient stress and culturing methods on toxin production and photosynthetic performance. Results showed substantial variability between clones in growth rates and toxicity levels under identical conditions. Toxin levels increased under silicon limitation and decreased with higher growth rates in chemostat experiments. Variable fluorescence measurements indicated nutrient stress impaired photosystem II and negatively correlated with toxin accumulation.
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)
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.
1. The document presents the discovery of JSI-124, a selective inhibitor of the JAK/STAT3 pathway, which was identified from screening a library of compounds for inhibition of STAT3 phosphorylation in cancer cells.
2. JSI-124 was found to reduce phospho-STAT3 levels in multiple human cancer cell lines and block STAT3 DNA binding and gene transcription, inhibiting tumor growth.
3. JSI-124 selectively targeted STAT3 signaling over other pathways and showed potent antitumor activity against human tumor xenografts in mouse models, demonstrating its potential as an anticancer therapeutic targeting the STAT3 pathway.
This document summarizes several studies that investigated methods for quantifying and tracking monocyte recruitment into atherosclerotic plaques. Initial studies in pigs used electron microscopy to observe monocyte movement into plaques. Subsequent studies labeled monocytes with fluorescent tags or genetic markers and tracked their movement into plaques in animal models like pigs, mice and rabbits. These studies found that labeled monocytes were detected within plaque lesions and that antibodies blocking adhesion molecules reduced monocyte recruitment, demonstrating the methods effectively tracked and quantified monocyte infiltration into plaques.
1) Several studies have traced the recruitment of monocytes into atherosclerotic plaques using labeling techniques. Gerrity et al labeled monocytes with FITC and found them adhered to plaque sites.
2) Willerson et al labeled macrophages with fluorescent microspheres and found them adhering to plaques. Antibodies to ICAM-1 and integrin reduced recruitment.
3) Steinberg et al transfused leukocytes between genetically different rabbits to track recruitment to plaques using PCR, finding over 600 donor cells per million in early fatty streaks and over 3,800 in advanced plaques.
Integrin V can form heterodimers with several subunits to mediate cell-cell and cell-extracellular matrix interactions. During zebrafish gastrulation, V is expressed maternally and zygotically. Here, we used a morpholino-mediated V knockdown strategy to study V function. Although V morphants displayed vascular defects, they also exhibited left-right body asymmetry defects affecting multiple visceral organs. This was preceded by mislocalization of dorsal forerunner cells (DFCs) and malformation of the Kupffer’s vesicle (KV) laterality organ
Commercial Application of Anoectochilus formosanus: Immunomodulating ActivitiesCây thuốc Việt
Anoectochilus formosanus is an important ethnomedicinal plant of Taiwan. We investigated the effect of oral administration of A. formosanus effective fraction (AFEF) on the innate immune response in mice. Male BALB/c mice were treated orally for 2 weeks with 500, 1000 and 1500 mg/kg of AFEF. Primary peritoneal macrophage harvest from mice that administered with AFEF (500 –1500 mg/kg) was directed to activate phagocytosis. AFEF significantly increased interferon-production from lymph node cells by ConA stimulation for 48 hours in AFEF (1500 mg/kg) treated group. AFEF might be the active fraction in activation of innate immunity.
1) Transgenic fish models carrying bacteriophage λ and plasmid pUR288 vectors were developed to improve methods for assessing health risks from environmental mutagens and establish new animal models for studying in vivo mutagenesis.
2) The bacteriophage λ transgenic medaka model uses the cII and lacI genes as mutational targets, allowing detection of mutations through packaging of the phage vector and infection of E. coli. Spontaneous mutation frequencies in medaka were comparable to rodent models.
3) Exposure to chemical mutagens like ENU induced concentration-dependent, tissue-specific, and time-dependent increases in cII mutations, demonstrating the utility of the transgenic fish model for studying mut
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 document describes two experiments that examined the effects of dietary zinc source and coccidial vaccine exposure on zinc homeostasis and immune status in broiler chickens. The experiments found that coccidial challenge decreased intracellular zinc levels and phagocytic capacity in the jejunum, while increasing phagocytic capacity in the caecal tonsils. Coccidial challenge also increased the ratio of zinc import to export transporters. Dietary zinc source had little impact except on one zinc transporter. The results suggest that during coccidial challenge, intestinal cells attempt to compensate for the drop in intracellular zinc by upregulating zinc import transporters.
The experiment aimed to determine if inserting arginine-producing enzymes from Chlorella vulgaris into a mutated form of Chlamydomonas reinhardtii lacking the arg2 gene would allow the mutated algae to grow. Arginine-producing enzymes were extracted from C. vulgaris and added to growth medium containing the mutated C. reinhardtii. There was growth of the mutated C. reinhardtii with the inserted enzymes without arginine supplements, supporting the hypothesis that the enzymes would allow the algae to produce arginine and grow. Control groups of C. reinhardtii grown with and without supplemental arginine showed different growth patterns.
Widespread Natural Occurrence of Hydroxyurea in AnimalsMichelle Pyle
This research article reports the discovery of naturally occurring hydroxyurea, an antibiotic compound, in many animal species across taxonomic groups. The highest levels were found in the little skate shark, with concentrations in some tissues high enough to have antiviral and anticancer effects based on in vitro studies. Embryos of the little skate also showed increasing hydroxyurea levels with development, indicating the ability to synthesize it. While levels in most other species examined were lower, certain tissues in some invertebrates and vertebrates also had concentrations expected to have antiviral effects. The widespread natural presence of hydroxyurea suggests it may play a role in disease resistance as part of the innate immune system in combating infections.
6 magalhães et al 2008 a hyaluronidase from potamotrygon motoro (freshwaterpryloock
1) Researchers purified and characterized a hyaluronidase enzyme from the venom of the freshwater stingray Potamotrygon motoro.
2) A two-step purification process involving gel filtration chromatography and ion exchange chromatography resulted in a 366-fold purification with a 79 kDa hyaluronidase enzyme.
3) The purified hyaluronidase had a pH optimum of 4.2 and maximum activity at 40°C. Its activity was inhibited by certain metal ions and heparin.
This document summarizes a study that investigated how mechanical forces applied to integrin receptors control intracellular signaling in osteoblasts. The researchers found that cyclic forces applied to the beta-1 integrin subunit at 1 Hz were more effective at stimulating calcium responses in osteoblasts than continuous forces. Cyclic forces also induced increased tyrosine phosphorylation of cytoskeleton-anchored proteins and greater activation of focal adhesion kinase and mitogen-activated protein kinase compared to continuous forces. These responses depended on an intact cytoskeleton and the presence of intracellular calcium. Analysis of spatial calcium signals revealed they originated near the stressed receptors, indicating cells can sense local stress via integrins.
IOSRPHR(www.iosrphr.org) IOSR Journal of Pharmacyiosrphr_editor
This document summarizes research on developing polyclonal antibodies to identify isoflavones in different legume species. Key points:
- Researchers produced polyclonal antibodies in rabbits to detect isoflavones. ELISA tests using these antibodies could quickly screen plants for isoflavone content.
- Seed samples from 8 legume species were tested for isoflavones using the ELISA. All species showed higher isoflavone levels than soybean (the typical isoflavone source), indicating other legumes may be alternative isoflavone sources.
- Extraction methods including hydrothermal treatment at 50°C increased levels of active aglycone isoflavone forms compared to no treatment.
- The research aims to characterize legume
Biodiversity conservation has conventionally dealt with management aspects, but over the last decade the issue of governance, i.e. who decides and how, has gained prominence, resulting in crucial paradigm shifts in protected area and other conservation policies and practice.
The document discusses the key components of language construction:
1) Primitives are the smallest units of meaning that cannot be broken down further, such as morphemes in natural languages.
2) Means of combination are rules for building words from primitives and phrases/sentences from words.
3) Means of abstraction allow assigning a simple name to a complex entity, such as pronouns in English that take on meaning based on context.
Este documento explica los cuatro casos gramaticales en alemán (nominativo, dativo, acusativo y genitivo) y cómo se relacionan con las funciones gramaticales en español como sujeto, complemento indirecto, complemento directo y complemento del nombre. También incluye una tabla detallada que muestra cómo declinar sustantivos, adjetivos y pronombres posesivos en alemán según el caso, género, número y terminación.
Este documento proporciona una introducción a los gestores de contenidos. Define qué son los gestores de contenidos y explica que proveen un entorno para actualizar, mantener y ampliar sitios web de manera colaborativa. Luego describe los tipos de gestores de contenidos existentes, incluyendo foros, blogs, wikis y portales. Finalmente, discute los roles de usuario y administrador y los gestores de contenidos más aplicados en el sector agropecuario.
U.S. Marine Emissions Regulations and Compliance Initiatives & Assessments, T...Team Finland Future Watch
The document summarizes U.S. marine emissions regulations and compliance initiatives. It finds that the U.S. has significant maritime emissions and the EPA's 2012 rule enforces IMO standards in U.S. coastal waters. Key compliance strategies are exhaust scrubbers, Tier 3 engine standards, and distillate fuel. States like California have additional rules, and initiatives encourage technologies like LNG, shore power, hybrids, and fuel cells to reduce emissions.
The document explores how one's perception of themselves can shape their reality. It presents hypothetical scenarios where the speaker perceives themselves as different entities such as a king, panther, mountain, and more. In each case, the speaker describes how if they perceived themselves in that way, that is what they would become, such as becoming a menacing panther if they perceived themselves as such. It ultimately concludes that no matter the hurdles, if one perceives themselves as living, then they are indeed alive.
The document discusses advertising a home daycare business. It recommends using word of mouth by telling everyone you know on social media and asking them to spread the word. It also suggests installing a sign with your information in the yard or putting a vinyl banner on the porch. The sign should include details about your program, experience, home, hours, rates, and a website or phone number for more information.
The document outlines the various advisory services provided by CohnReznick, including:
- Valuation advisory for financial reporting, taxes, transactions, real estate, and complex instruments.
- Governance, risk and compliance services such as internal audits, IT audits, risk management, and fraud assessments.
- Management consulting services like CFO advisory, process improvement, strategic planning, and supply chain optimization.
- Technology and digital services including IT strategy, data analytics, digital solutions, and cloud computing.
- Transaction advisory and M&A consulting for deal evaluation, due diligence, integration, and dispute resolution.
- Cybersecurity services to assess risks and mitigate threats through strategies, assessments, and response
El documento describe la estructura y propiedades básicas de los átomos. Explica que un átomo está compuesto de un núcleo central con protones y neutrones, rodeado por electrones. También describe las fuerzas que mantienen unidos los componentes del átomo y los diferentes modelos atómicos desarrollados.
Communicating effectively to advance street safety is not a new goal, but Vision Zero is bringing greater urgency and critical thinking to this need. It also brings together a wider and more diverse range of stakeholders who recognize the value of well-planned, measureable communication efforts. The language of Vision Zero itself -- with the goal to eliminate all traffic fatalities and severe injuries -- communicates a more ambitious approach to street safety and rests on the basic understanding that these serious losses are preventable.
In this case study, we look at two early-adopter cities’ -- New York City and San Francisco -- promising approaches to communicating about Vision Zero in order to garner attention and influence behavior -- at all levels of society.
El estudiante encontró más difícil entender las unificaciones de Alemania, Italia y las revoluciones de 1830 y 1848. Valoró el funcionamiento de su grupo de trabajo con un 7,5 sobre 10, señalando que estaban unidos y se ayudaban, aunque podrían haber mejorado en las explicaciones y tareas. Lo más complicado fue subir la entrevista al blog, pero no tuvo dificultades con ninguna otra herramienta. Aprendió mucho sobre Napoleón y Robespierre al preparar la entrevista sobre este último. No tuvo problemas con ningún t
Epidemiologi PM 10 pada kabut asap akibatkan ISPANida Salamah
Makalah ini membahas tentang kabut asap di Kota Banjarbaru yang diakibatkan oleh kebakaran lahan gambut dan hutan. Kabut asap mengandung partikel-partikel berbahaya seperti PM-10 yang dapat menyebabkan penyakit infeksi saluran pernafasan atas (ISPA) terutama pada anak-anak. Penelitian menunjukkan peningkatan kasus ISPA di Kota Banjarbaru selama musim kemarau akibat peningkatan konsentras
El documento lista nombres de figuras históricas y políticas importantes de Colombia entre 1972 y 2016, incluyendo Simón Bolívar, Francisco de Paula Santander, Policarpa Salavarrieta, y Gabriel García Márquez. Muchas de las figuras se repiten a lo largo de la lista.
KNIME Analytics Platform is an open source data analytics platform that allows users to discover insights from data through customizable workflows. It provides over 1000 analytic techniques through nodes for tasks like statistics, data mining, text mining, and more. Workflows can integrate various data sources, transform the data, apply models, and output results in standard formats. The platform is open source and free to use, customize, and extend through its community and commercial extensions.
Windows Forms is a framework for building desktop applications for Windows using .NET. It allows developers to easily create graphical user interfaces (GUIs) for software applications and is included in the Visual Studio integrated development environment (IDE). Windows Forms supports drag-and-drop design and a code editor that allows applications to be built visually or through code.
The Actionable Guide to Doing Better Semantic Keyword Research #BrightonSEO (...Paul Shapiro
1) Semantic search relies on understanding the conceptual relationships between keywords rather than exact matches, so SEOs must conduct more thorough semantic keyword research.
2) Tools like KNIME allow SEOs to automate data collection from sources like search engines and social media, analyze the data using techniques like TF-IDF and LDA to group keywords semantically, and visualize relationships to guide on-page optimization.
3) By understanding conceptual topics and how consumer language is used, SEOs can better optimize websites for searcher intent to perform well in semantic search.
Magalhães, et al. 2006. biological and biochemical properties of the brazilianpryloock
This document summarizes a study on the biological and biochemical properties of venoms from two species of Brazilian freshwater stingrays, Potamotrygon cf. scobina and Potamotrygon gr. orbignyi. The study found that both venoms induced edema, pain responses, and increased leukocyte activity in mice. Venom effects were reduced when heated. Injection of venoms caused tissue necrosis, low protease activity, and no hemorrhaging. Coinjection of venom and mucus secretion caused more vigorous necrosis. The study provides evidence of toxic effects for both stingray venoms.
This document summarizes several genes and proteins of interest in entomological research, including Bt Cry toxins, plant metabolites, enzyme inhibitors, plant lectins, insect hormones, and genes involved in insect sex determination and development. It also discusses juvenile hormone and its role in insect metamorphosis, as well as neuropeptides and their functions in insect behavior, physiology, and homeostasis. Finally, it covers protease inhibitors from plants that play a role in insect control by inhibiting insect digestive enzymes.
Identification of fish species using dna barcode from visakhapatnam, east coa...RUSHINADHA KAKARA
This document describes a study that generated DNA barcodes for fish species found at a fishing harbor in Visakhapatnam, India. DNA was extracted from tissue samples of 50 fish individuals representing different species. The cytochrome c oxidase subunit I (COI) region of the mitochondrial DNA was amplified and sequenced. The resulting DNA barcodes were analyzed using bioinformatics tools including BLAST searches and multiple sequence alignments. A phylogenetic tree was constructed to examine relationships between species. One objective was to investigate potential misidentification of Tripletail fish and develop a reference barcode library for species identification in the study area.
Identification of fish species using dna barcode from visakhapatnam, east coa...RUSHINADHA KAKARA
This document describes a study that generated DNA barcodes from fish species collected at a fishing harbor in Visakhapatnam, India. DNA was extracted from tissue samples of 50 fish individuals and a 658 base pair region of the COI gene was amplified and sequenced. The sequences were analyzed using tools like ORF finder, BLAST, and multiple sequence alignment. A phylogenetic tree was constructed to investigate relationships between sequences. The goals were to create a reference barcode library for the region and investigate species identification and potential cryptic species. The study focused on analyzing barcodes of Tripletail fish, which previous work on barcoding this species is limited.
This document summarizes a study that examined the antimicrobial properties of mucus from the chame fish (Dormitator latifrons). The study found inhibitory effects of chame mucus against several bacteria strains. Specifically, chame mucus showed inhibitory effects against 2 out of 3 Bacillus strains tested as well as strong inhibitory effects against Vibrio vulnificus and Vibrio harveyi. A lower level of inhibition was also observed against Vibrio anguillarum. The results suggest the presence of antibacterial agents in chame fish mucus, which could potentially be applied to animal and human health.
The document summarizes a study that investigated the effects of three Mexican medicinal plants (Psacalium decompositum, Psacalium peltatum, and Acourtia thurberi) on blood glucose levels. The study found that:
1) Decoctions of P. decompositum and A. thurberi significantly reduced blood glucose levels in healthy mice, while P. peltatum decreased levels to a lesser extent.
2) Decoctions of all three plants diminished hyperglycemia in rabbits made temporarily hyperglycemic, with P. decompositum showing the greatest effect, lowering peak blood glucose levels by 17.1%.
3) Preliminary chemical analysis identified
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
Lyciumbarbarum (Gojji berry) belongs to family Salonaceae which is found in China and Himalayan. This herb is used to prevent various diseases and in medical treatments as an alternative medicine being widely used for its antioxidant and revitalizing potential effects. In recent years, Gojji has become increasingly popular in Europe and North America as a "superfruit" and dietary supplement. The belief that herbal products do not bring any risk to health, is part of popular culture. However the term "natural" assigned to many products cannot assure no health risk. The aim of this study was to evaluate the possible genotoxic effects of aqueous extract of Lyciumbarbarum (Gojji berry) by micronucleus test and comet assay. Thirty Rattus norvegicus were divided into three equal groups: 1) experimental group, submitted to Gojji berry (200mg/kg orally); 2) positive control group (cyclophosphamide), and; 3) negative control group (distilled water). Micronucleus Tests were done by smear method of bone marrow cells performed after 48h for acute, and 72h for chronic exposure. The comet assay was performed on peripheral blood taken from the tail of each animal 4h, and 24h after intervention. Cytotoxicity was assessed by observing the DNA damage measuring the percentage of DNA in the tail (% DNA- measurement of the proportion of the total DNA present in the tail) and the tail moment (TM-tail length times the percentage of DNA in the tail), calculated by 100 nucleoids per animal and the presence of micronuclei in 2,000 polychromatic erythrocytes per animal. Analysis of variance (ANOVA) followed by Tukey test at 5% significance was used comparing the results. The data showed no significant difference in the frequency of DNA damage and the number of micronuclei between the experimental group and the negative control group. The results also suggest that the aqueous extract of Lyciumbarbarum (Gojji berry) at the dose of 200 mg/kg showed no genotoxic effect, which could, to a certain point, justifies its use.
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
Lyciumbarbarum (Gojji berry) belongs to family Salonaceae which is found in China and Himalayan. This herb is used to prevent various diseases and in medical treatments as an alternative medicine being widely used for its antioxidant and revitalizing potential effects. In recent years, Gojji has become increasingly popular in Europe and North America as a "superfruit" and dietary supplement. The belief that herbal products do not bring any risk to health, is part of popular culture. However the term "natural" assigned to many products cannot assure no health risk. The aim of this study was to evaluate the possible genotoxic effects of aqueous extract of Lyciumbarbarum (Gojji berry) by micronucleus test and comet assay. Thirty Rattus norvegicus were divided into three equal groups: 1) experimental group, submitted to Gojji berry (200mg/kg orally); 2) positive control group (cyclophosphamide), and; 3) negative control group (distilled water). Micronucleus Tests were done by smear method of bone marrow cells performed after 48h for acute, and 72h for chronic exposure. The comet assay was performed on peripheral blood taken from the tail of each animal 4h, and 24h after intervention. Cytotoxicity was assessed by observing the DNA damage measuring the percentage of DNA in the tail (% DNA- measurement of the proportion of the total DNA present in the tail) and the tail moment (TM-tail length times the percentage of DNA in the tail), calculated by 100 nucleoids per animal and the presence of micronuclei in 2,000 polychromatic erythrocytes per animal. Analysis of variance (ANOVA) followed by Tukey test at 5% significance was used comparing the results. The data showed no significant difference in the frequency of DNA damage and the number of micronuclei between the experimental group and the negative control group. The results also suggest that the aqueous extract of Lyciumbarbarum (Gojji berry) at the dose of 200 mg/kg showed no genotoxic effect, which could, to a certain point, justifies its use.
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
This document summarizes a study that evaluated the genotoxic effects of an aqueous extract of Goji berry (Lyciumbarbarum) using micronucleus and comet assays in rats. Rats were divided into three groups: an experimental group that received 200 mg/kg of the Goji berry extract orally, a positive control group that received cyclophosphamide, and a negative control group. The comet assay showed no significant increase in DNA damage in the experimental group compared to the negative control group at 4 or 24 hours. The micronucleus test also found no significant difference in micronuclei between the experimental and negative control groups for acute or chronic exposure. The results suggest that the Goji berry
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
The document describes a study that found that exposure to the common herbicide atrazine at low doses caused sexual abnormalities in frogs, including hermaphroditism and demasculinization. Larvae exposed to more than 0.1 parts per billion of atrazine developed as hermaphrodites, and males exposed to over 1 part per billion had demasculinized larynges and decreased testosterone levels. The study suggests that atrazine exposure disrupts hormone production and may explain some cases of declining amphibian populations globally.
Studies on cellulose degrading microorganisms associated with rumen of rumina...Premier Publishers
Studies on cellulose degrading microorganisms associated with rumen of ruminants was carried out from ruminants (ram, cow, and goat), through culture, microscopic identification, Biochemical test and cellulose degrading methods. In the rumen content of ram four bacteria were isolated Bacteriodes and Staphylococcus had the highest percentage (33.3%) each while Veillonella and Bacillus had 16.6% each. Seven bacteria were isolated from cow with Streptococcus having (22.2%) Staphylococcus (22.2%), while Bacteroides, Yersinia, Peptococcus, Nesseria and Bacillus had equal distribution. Goat had eight bacteria including Bacteroides, Clostridium, Yersinia, Staphylococcus, Homofermentative Lactobacillus Alcaligens and Bacillus all of which had equal distribution. Bacteroides and Bacillus are common in all rumens, with Bacteroides, being more prevalent in the ram. study revealed that ruminants harbors various organisms that are active cellulose degraders, out of which Bacteroides specie grow best on cellulose agar. For fungi, Aspergillus flavus and Aspergillus fumigatus highly degrades cellulose, Scopulariopsis candida degrades minimally. The study revealed that ruminants harbors various organisms that are active cellulose degraders, out of which Bacteroides specie grow best on cellulose agar. Therefore, Rumen should be used as a site for isolation of micro organisms capable of cellulose hydrolysis in order to reduce the coast of purchasing commercial enzymes
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
ABSTRACT- Some Lactobacillus species (L. acidophilus, L. casei and L. plantarum) were isolated from locally fermented products (ogi, fura de Nunu and wara) and their effect on microbial infections caused by some pathogenic bacteria (E.coli, K. pneumoniae, Pseudomonas aeruginosa and Staphyloccoccus aureus) isolated from urine and high vaginal swab samples were studied using standard micriobiological methods.Fifiteen (15) healthy guinea pigs used for the study were divided into three (3) groups of five (5) guinea pigs each and placed in three (3) different cages. The pigs were initially fed for two (2) weeks (acclimatization period) with conventional feeds before administering the treatment. Lactobacillus species were introduced into the guinea pigs in cage 2 after the acclimatization period. Subsequently, the guinea pigs in cages 1 and 2 were orally infected with all the clinical bacteria pathogens while the guinea pigs in cage 3 which served as control were left with no microbial treatment. Ten (10) days after treatment, the packed cell volume (PCV), haemoglobin concentration (HBC), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity level were determined. Striking differences were observed from guinea pigs in the different cages. The effectiveness of Lactobacilli (probiotics) was evident when the guinea pigs in cages 1 and 2 were compared. The guinea pigs in cage 1 which were infected with pathogens but no probiotics had lower blood level (mean PCV= 24.8%) and inferior liver condition (mean ALT=58.18µl; mean AST=51.91µl). Higher blood level (Mean PCV=45%) and superior liver conditions (Mean ALT=9.51µl; mean AST=9.7µl) were obtained for guinea pigs in cage 2 which were infected with the same pathogens and fed with probiotics. The control (cage 3) had the highest PCV level and best liver conditions (mean PCV=46.6%, means ALT= 7.65µl; mean AST=11.83µl).Th .This might be attributed to the fact that they were not infected with pathogenic organisms. Lactobacillus species administered are promising probiotics against the tested bacterial pathogens.
Keywords: Lactobacillus species, Guinea pig, Bacteria pathogen, Enzymes assay, Haematological Parameters, Probiotics
- Listeria was first isolated in 1926 from infected rabbits and named Bacterium monocytogenes. It was later renamed Listeria in 1940.
- Listeria monocytogenes is a pathogenic bacteria that can cause listeriosis in humans and animals through the consumption of contaminated food. It is particularly associated with consumption of contaminated silage by cattle and sheep.
- Symptoms of listeriosis in animals include dullness, circling behavior, head tilt, facial paralysis, and in some cases abortion. It primarily infects the central nervous system.
The document discusses various adaptations parasites have evolved to survive in their hosts. It describes physiological adaptations like asexual reproduction, hermaphroditism, and high fecundity that allow parasites to rapidly increase their numbers. Behavioral adaptations help parasites find and transmit between hosts, like periodic larval emergence. Structural adaptations enhance attachment and penetration of hosts, such as suckers and hooks. Many parasites also modify host behaviors to aid transmission.
This study tested different cryopreservation methods for sperm of two endangered mahseer fish species: Tor khudree and T. putitora. It found that a combination of 9% dimethyl sulfoxide and 11% glycerol cryoprotectants, with a 30 minute equilibration period and freezing at 8 cm above liquid nitrogen for 10 minutes, produced the highest sperm motility and hatching rates after thawing for both species. This protocol provides a standardized method for cryopreserving and storing the genetic material of these threatened fish, aiding conservation efforts.
This study examined the parasites of the Señorita Wrasse fish in the Santa Barbara kelp forests. A total of 12 fish were dissected and 7 groups of parasites were found, including nematodes, trematodes, cestodes, and acanthocephalans. Most parasites were found as larvae, indicating the fish acts as an intermediate host. Nematodes had the highest prevalence and intensity. Several species of trematodes, cestodes, and acanthocephalans were also found. The identification of these trophically transmitted parasites provides insights into possible predator-prey relationships in the kelp forest food web, with the fish linking lower trophic levels like crustaceans to higher
1. The document discusses how zoology provides an essential foundation for understanding modern biological research, like genomics and parasitic diseases. It gives examples of how studying the taxonomy and biology of schistosomes and body lice has helped answer interesting evolutionary questions.
2. It then focuses on schistosomes, parasitic flatworms that cause schistosomiasis. Genomic studies have helped reveal genes involved in the parasite's complex lifecycle and ability to infect different hosts. However, drug resistance requires identifying new drug targets.
3. It also discusses how genomic analysis of the body louse determined that humans began regularly wearing clothing, by tracing the evolutionary divergence of head and body lice.
1. Journal of Invertebrate Pathology 87 (2004) 123–128
www.elsevier.com/locate/yjipa
0022-2011/$ - see front matter 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.jip.2004.09.001
Leishmania parasites (Kinetoplastida: Trypanosomatidae) reversibly
inhibit visceral muscle contractions in hemimetabolous
and holometabolous insects
Rajeev Vaidyanathan¤
Department of Parasitology, Hadassah Medical School, Hebrew University, Ein Kerem, P.O. Box 12272, Jerusalem 91120, Israel
Received 30 June 2004; accepted 1 September 2004
Available online 5 November 2004
Abstract
Female sand Xies can acquire protozoan parasites in the genus Leishmania when feeding on an infected vertebrate host. The para-
sites complete a complex growth cycle in the sand Xy gut until they are transmitted by bite to another host. Recently, a myoinhibitory
peptide was isolated from Leishmania major promastigotes. This peptide caused signiWcant gut distension and reversible, dose-depen-
dent inhibition of spontaneous hindgut contractions in the enzootic sand Xy vector, Phlebotomus papatasi. The current study further
characterizes myoinhibitory activity in L. major and other kinetoplastid parasites, using the P. papatasi hindgut and other insect
organ preparations. Myoinhibitory activity was greatest in cultured promastigotes and in culture medium in late log-phase and early
stationary-phase, coinciding with development of infective Leishmania morphotypes in the sand Xy midgut. L. major promastigote
lysates inhibited spontaneous contractions of visceral muscle preparations from hemimetabolous (Blattaria and Hemiptera) and
holometabolous (Diptera) insects. Inhibition of visceral muscle contractions in three insect orders indicates a conserved mode of
action. Myoinhibitory activity was detected also in Leishmania braziliensis braziliensis, a Sudanese strain of Leishmania donovani, and
the kinetoplastid parasite Leptomonas seymouri. Protozoan-induced myoinhibition mimics the eVect of insect myotropins. Inhibiting
host gut contractions protects Leishmania parasites from being excreted after blood meal and peritrophic matrix digestion, allowing
development and transmission of infective forms.
2004 Elsevier Inc. All rights reserved.
Keywords: Host–parasite interactions; Vector competence; Sand Xy; Leishmania; Leptomonas; Herpetomonas; Myoinhibition
1. Introduction
The sand Xy Phlebotomus papatasi (Diptera: Psychod-
idae) is the sole vector of Leishmania major (Kinetop-
lastida: Trypanosomatidae) to humans in arid foci of
enzootic cutaneous leishmaniasis (Peters and Killick-
Kendrick, 1987; Schlein et al., 1982). Female sand Xies
ingest macrophages containing L. major amastigotes
when they bite an infected vertebrate host. Blood is
encased in a chitinous peritrophic matrix (PM) in the
gut, wherein amastigotes divide repeatedly and trans-
form into a uniform population of promastigotes
(Blackburn et al., 1988). The PM disintegrates after
blood meal digestion, and free parasites may be excreted
along with PM and blood meal remnants.
Several mechanisms facilitate parasite persistence in
the gut after blood digestion. Promastigotes may anchor
their Xagella between midgut microvilli (Adler and The-
odor, 1926; Killick-Kendrick et al., 1974; Warburg et al.,
1986, 1989). Polymorphisms in Leishmania lipophospho-
glycan (LPG), the main surface glycoconjugate, mediate
*
Fax: +1 530 752 1537, Present address: Department of Entomolo-
gy, Briggs Hall, Shields Avenue, University of California, Davis, CA
95616, United States..
E-mail address: rvaidyanathan@ucdavis.edu.
2. 124 R. Vaidyanathan / Journal of Invertebrate Pathology 87 (2004) 123–128
promastigote binding, release, and anterior migration
(Pimenta et al., 1992; Sacks and Kamhawi, 2001; Sacks
et al., 1994, 1995). However, a large population of unat-
tached parasites persists in the gut lumen (Walters et al.,
1987, 1989), and recently a mechanism was proposed for
how these parasites might remain in the gut. A 12 kDa
peptide isolated from L. major promastigotes was shown
to reversibly inhibit hindgut contractions and cause sig-
niWcant gut distension in P. papatasi, thereby protecting
unattached parasites from expulsion after blood diges-
tion (Vaidyanathan, in press).
The current study further characterizes myoinhibi-
tory activity in L. major and other kinetoplastid para-
sites. Stage-speciWc expression of L. major myoinhibitory
activity in cultured promastigotes and conditioned cul-
ture medium was detected using the P. papatasi hindgut
assay. Contractions of hindgut, dorsal blood vessel and
oviduct preparations from other insect species were also
inhibited by L. major promastigote lysate. Four other
species of Leishmania and three other species of obligate
gut parasites of insects were tested for their ability to
inhibit P. papatasi hindgut contractions.
2. Materials and methods
2.1. Parasite cultures
Leishmania major MHOM/IL/86/Blum (Jordan Val-
ley strain), Leishmania braziliensis braziliensis MHOM/
BR/75/M2903, Leishmania infantum MCAN/IL/L760,
Leishmania tropica MHOM/IL/L590, Leishmania dono-
vani (Khartoum and DD8 Indian strains), Herpetomo-
nas muscarum (a kinetoplastid parasite of house Xies),
and Leptomonas seymouri (a kinetoplastid parasite of
lygaeid bugs) were obtained from the W.H.O. Leish-
mania Reference Center, the Kuvin Centre for the
Study of Infectious and Tropical Diseases, Jerusalem.
All reagents and protease inhibitors were purchased
from Sigma (Rehovot, Israel), unless otherwise speci-
Wed. Parasites were grown in Dulbecco’s modiWed
Eagle’s medium (DMEM) (Biological Industries [B.I.],
Beit Haemek, Israel) with high glucose content, 10%
heat-inactivated fetal calf serum (FCS) (B.I.), 4 mM L-
glutamine, 2 mM adenosine, and 2% (v/v) Wlter-steril-
ized human urine. Cultures were grown at 28 °C and
passaged every 4 days. Cultures of Crithidia fasciculata,
a kinetoplastid gut parasite of mosquitoes, were grown
at 28 °C in brain–heart infusion (B.I.) and passaged
daily.
Homogenate preparation was the same for all para-
sites. Late log-phase promastigote cultures with high
parasite density (107
–108
parasites/ml) were spun at
2000g for 10 min at 8 °C and washed twice with ice-cold
Aedes aegypti buVered saline (ABS, Wnal concentra-
tions: 0.6 mM MgCl2, 4.0 mM KCl, 1.8 mM NaHCO3,
150 mM NaCl, 25 mM Hepes–NaOH, and 1.7 mM
CaCl2, pH 7.4). A protease inhibitor cocktail was added
to wet parasite volume to inhibit autolysis (Wnal
concentrations: 1.0 mM AEBSF, 0.5 mM EDTA, 65 M
bestatin, 7 M E-64, 0.5 M leupeptin, and 0.15 M
aprotinin). To lyse parasites, cell pellets were freeze–
thawed three times in liquid N2 and at 30 °C. Crude
homogenates were spun at 12,000g for 30 min.
Concentrations of supernatant proteins were assayed
by the Bradford method (Bradford, 1976). Based on
previous results, only lysate supernatant was used for
bioassays.
2.2. Stage-speciWc expression of myoinhibitory activity
Protein samples from culture medium used for L.
major growth were precipitated with 85% of saturation
(NH4)2SO4 at 0 °C. Both precipitate and supernatant
were dialyzed (3500 Da MWCO, Spectra/Por Mem-
brane, Spectrum Labs, Jerusalem) for 24 h at 4 °C
against ABS decreased to 50 mM NaCl, and tested using
the sand Xy hindgut bioassay (below). Parasites and
medium (four replicates) were harvested from day 1 to 7
of growth, and daily parasite density was calculated.
Cultures were centrifuged at 2000g for 10 min at 8°C
and washed twice with ABS. Parasites were lysed as
described above, and crude homogenates were stored at
¡70 °C until bioassayed for stage-speciWc expression of
the paralytic factor. For culture medium and parasite
lysates, a range of 10–100 g/ml proteins were tested, and
activity was standardized as percent inhibition per
microgram proteins.
2.3. Experimental insects
Colonies of the sand Xies P. papatasi (originated from
Kfar Adumim, 10 km east of Jerusalem) and Lutzomyia
longipalpis (Bahia strain) were maintained according to
Modi and Tesh (1983). Insectary conditions were tem-
perature of 26 §1 °C, relative humidity 80% and a 17:7
light:dark cycle. Two- to six-day-old sugar fed sand Xies
were used for all experiments. The Rockefeller strain of
A. aegypti (Diptera: Culicidae) was maintained under
the same conditions.
The blood-feeding bug Rhodnius prolixus (Hemiptera:
Reduviidae) was a gift from J.M.C. Ribeiro and main-
tained on rabbit blood under the same conditions; sec-
ond instars were used in all experiments. Cockroaches
(Blattaria: Periplaneta americana and Supella longipalpa)
were kept at room temperature, with pulverized cat food
and water. The sheep blowXy Lucilia sericata (Diptera:
Calliphoridae) was donated by K.Y. Mumcuoglu and
maintained on 10% sucrose and beef liver for oviposi-
tion. Flies (Diptera) in the families Sarcophagidae, Psy-
chodidae, and Muscidae were caught at natural resting
sites and maintained on 10% sucrose.
3. R. Vaidyanathan / Journal of Invertebrate Pathology 87 (2004) 123–128 125
2.4. Insect bioassays
Whole guts from male and female P. papatasi were
dissected into a watch glass with 90–99 l oxygenated
ABS warmed to 30 °C and allowed to recover until
hindgut contractions stabilized. Hindgut contractions
were counted for 5 min increments. Samples of proteins
from culture medium or from parasite lysate were
added 5 min after the hindgut stabilized and contracted
regularly. Values for activity were standardized as
percent inhibition of P. papatasi hindgut contractions
(relative to 5 min equilibration) per microgram pro-
teins.
Organs of other insects were dissected into either
Cockroach buVered saline (CBS, Wnal concentrations:
150mM NaCl, 5.0mM KCl, 5.0mM CaCl2, 3.0mM
MgCl2, 4.0mM Hepes–NaOH, and 40mM glucose, pH
7.2) (Lange et al., 1993) or an insect Ringer’s solution
(Wnal concentrations: 154mM NaCl, 2.7mM KCl,
1.8mM CaCl2, 12.0mM Hepes–NaOH, 22.0mM glucose,
pH 7.0) (Cook et al., 1975; Duve et al., 1999). BuVer vol-
ume depended on the size of the preparation. As with the
sand Xy hindgut bioassay, other insect tissues were
allowed to equilibrate and their contractions counted for
1–5min before addition of L. major lysate. Contractions
were counted further for 1–5min intervals. Inhibition was
calculated as the number of contractions after addition of
lysate proteins relative to the number of contractions dur-
ing the equilibration period. A range (1.2–24 g/ml) of
protein concentrations was tested, and activity was stan-
dardized as percent inhibition per microgram protein.
Reversibility of inhibition was tested using prepara-
tions of P. americana hindgut, Sarcophaga dorsal blood
vessel, and A. aegypti oviduct. Treated preparations were
rinsed twice in CBS or ABS and returned to warm,
untreated buVer.
3. Results and discussion
3.1. Stage-speciWc expression
A previous study showed that a unique peptide was
produced by L. major promastigotes in vitro, which
reversibly inhibited P. papatasi hindgut contractions and
resulted in gut distension (Vaidyanathan, in press). In
the current study, myoinhibitory activity was detected in
L. major lysate proteins on days 2–7 of growth, with
peak activity detected at late log-phase, early stationary-
phase (Fig. 1). By day 7, myoinhibitory activity had
decreased by 75% relative to peak activity on day 5.
Three days after blood feeding, the blood bolus has been
digested and voided. Maximal activity from cultured
promastigotes coincided with presence of free forms in
the sand Xy midgut lumen when nectomonads are vul-
nerable to peristaltic ejection.
Myoinhibitory activity in culture medium was detected
at the same time as its appearance in parasites (Fig. 1).
Peak activity in culture medium was detected at station-
ary-phase. Activity in the culture medium was higher than
in parasite lysate for days 6 and 7. Activity decreased by
day 7, the second day of stationary phase, a decrease of
20% relative to peak activity on day 5. Activity in both
sources was abolished by trypsin or proteinase-K, indicat-
ing that the active factor was a protein (results not
shown). During blood meal digestion and excretion, pro-
cyclic promastigotes transform into nectomonad forms in
the anterior abdominal midgut. Nectomonads may
migrate anteriorly to the thoracic midgut and stomodeal
valve, remain in the midgut lumen, or interdigitate their
Xagella between midgut microvilli. By day 7, slender nec-
tomonads transform into shorter haptomonads which
invade the Xy pharynx and esophagus (Schlein, 1993).
Most parasites have not migrated forward and remain
vulnerable to expulsion from the midgut. Although obser-
vations from cultured parasites might not reXect the situa-
tion in a sand Xy, if parasites and secreted proteins act
similarly in the gut, then myoinhibition would function
even after parasites are no longer secreting (or secreting
very little), ensuring that free forms in the lumen are not
voided. Moreover, maximal myoinhibitory activity after
blood digestion coincides with low midgut trypsin and
chymotrypsin levels, thereby avoiding myoinhibitory pep-
tide inactivation during proteolysis of blood (Borovsky
and Schlein, 1987; Ramalho-Ortigão et al., 2003).
In addition, the parasite population in sand Xies
includes several morphotypes not found in culture. Flies
remain infected for life, and some morphotypes might
continue secreting constantly.
3.2. Myoinhibition in diVerent insect preparations
Leishmania major lysate proteins were evaluated
for myoinhibitory activity on hindgut preparations of
Fig. 1. Inhibition of sand Xy hindgut contractions 5 min after applica-
tion of L. major proteins (10–100 g/ml) from parasites and precipi-
tated culture medium, harvested at 1–7 days of culture growth. Each
bar represents the mean of four trials.
0
1
2
3
4
5
6
7
8
1 2 3 4 5 6 7
Days
Percentinhibition
permicrogramproteins
0.0E+00
5.0E+06
1.0E+07
1.5E+07
2.0E+07
2.5E+07
Parasitesperml
Inhibition from parasite lysate
Inhibition from culture medium
Parasites per ml
4. 126 R. Vaidyanathan / Journal of Invertebrate Pathology 87 (2004) 123–128
P. americana, Psychoda sp., A. aegypti, L. sericata, S.
longipalpa, and R. prolixus. Hindguts of second instar P.
americana stopped contracting immediately upon appli-
cation of 65 g/ml L. major proteins. P. americana hind-
guts rinsed in CBS and returned to fresh CBS resumed
contracting (Fig. 2). Between 1 and 13% inhibition per
microgram L. major proteins (1.2–24 g/ml) was detected
against hindgut preparations of all insects tested (Fig. 3).
Highest activity per microgram lysate proteins was seen
in hindgut preparations of the sand Xies P. papatasi
(13%) and L. longipalpis (8.3%), both enzootic vectors of
Leishmania parasites. P. papatasi is the enzootic vector
of L. major; L. longipalpis, a South American sand Xy
species, transmits L. b. braziliensis, a peripylarian para-
site which has a stage of development in the hindgut
before establishing a midgut infection (Lainson and
Shaw, 1979).
To test for general inhibition against spontaneous vis-
ceral muscle contractions, L. major lysate proteins were
applied to dorsal blood vessels of Sarcophaga sp. and oviducts of A. aegypti. L. major proteins at a concentra-
tion of 24 g/ml immediately inhibited spontaneous con-
tractions of oviduct (59%) and dorsal blood vessel (22%)
preparations (Fig. 4). Surprisingly, in assays with A.
aegypti, inhibition of spontaneous oviduct contractions
was greater than inhibition of hindgut contractions.
The protein’s mode of action might be deduced from
these bioassays with insect visceral muscle. The myoin-
hibitory protein likely exerts its eVect through a com-
mon pathway of receptor–ligand interactions. The
enteric nervous system in arthropods releases neuropep-
tides which regulate visceral muscle activity (Coast and
Webster, 1998). Insect visceral muscle contraction and
relaxation are mediated by Ca2+
(Cook et al., 1975; Wil-
cox and Lange, 1995). Myosuppressins and other
myoinhibiting peptides in insects function by blocking
voltage-gated and ligand-gated Ca2+
channels in the
plasma membrane (Orchard et al., 1997). The Wrst
inhibitory neuropeptide isolated from an insect was
leucomyosuppressin, which inhibited spontaneous
contractions of the cockroach hindgut (Holman et al.,
1986). The tetrapeptides Phe–Met–Arg–Phe–amide
(FMRF-amide) and Phe–Leu–Arg–Phe–amide (FLRF-
amide) stimulate and inhibit cardiac and skeletal muscle
contractions in invertebrates (Cuthbert and Evans,
1989). FLRF-amides isolated from Locusta migratoria
inhibit locust heart rhythm, reduce spontaneous oviduct
contractions, and relax basal tone and amplitude of
hindgut contractions (Schoofs et al., 1993). Many other
insect neuropeptides interfere with contractions of gut
muscle and of dorsal blood vessel (Bendena et al., 1997;
Vilaplana et al., 1999).
Leishmania major lysates inhibited hindgut contrac-
tions of eight insect species and oviduct and dorsal blood
vessel contractions in Diptera. This conserved activity in
a spectrum of insects and insect organs may operate by
interfering with Ca2+
Xux in target tissues by blocking
Fig. 2. Reversible inhibition of P. americana hindgut contractions with
application of 65 g/ml L. major lysate proteins. Lysate was added to
hindgut preparations at 7 min, and allowed to act for 4 min. The hind-
gut was rinsed with CBS and returned to fresh saline at 11 min. Mean
values for Wve assays are shown.
Fig. 3. Means and standard deviations of percent inhibition by L.
major lysate proteins on isolated hindguts of diVerent insect species.
Protein concentrations varied among samples (1.2–24 g/ml), so all
values are standardized by percent inhibition per microgram lysate
proteins.
0
2
4
6
8
10
12
14
P. papatasi L. longipalpis Psychoda R. prolixus L. sericata S. longipalpa A. aegypti
Percentinhibitionpermicrogramproteins
Fig. 4. Reversible inhibition of spontaneous contractions of A. aegypti
oviduct (OVI) and Sarcophaga dorsal blood vessel (DBV) with appli-
cation of 24 g/ml L. major lysate proteins. Lysate was added to dis-
sected Xies at 6 min, and allowed to act for 4 min. Dissections were
rinsed with ABS at 10 min and allowed to recover. Means and stan-
dard deviations for three assays are shown.
0
25
50
75
100
1 3 5 7 9 11 13 15
Minutes
Percentinhibitionpermicrog.proteins
DBV
OVI
5. R. Vaidyanathan / Journal of Invertebrate Pathology 87 (2004) 123–128 127
Ca2+
channels and decreasing Ca2+
-dependent action
potentials, a mechanism shared by myotropic neuropep-
tides of insects. Further work is necessary to identify the
exact mode of action.
3.3. Myoinhibitory activity in diVerent kinetoplastid
parasites
Pathogenic eVects of trypanosomatids in their insect
hosts are well-documented (Schaub, 1992, 1994). Lysate
proteins (100 g/ml) of L. seymouri, L. donovani, L. b.
braziliensis, and L. major inhibited P. papatasi hindgut
contractions in increasing order of eVect (Fig. 5). The
Sudanese strain of L. donovani inhibited 20% of P.
papatasi hindgut contractions, and this eVect was
reversible (results not shown). The Indian strain had no
eVect. Growth media composition may profoundly
aVect enzyme secretion by trypanosomatid parasites
(Schlein and Jacobson, 1992). EVect of growth medium
likely explains diVerences in myoinhibitory expression.
Cultures of DMEM supplemented with 10% FCS were
used for all parasite species, and this could have inhib-
ited or masked paralytic peptide expression in diVerent
protozoa. Another explanation may be that diVerent
parasites have adapted to diVerent barriers in their
respective hosts, and testing these species with P. papat-
asi does not reXect their potential for persistence and
transmission. Further studies should include culturing
L. b. braziliensis in diVerent media and testing parasite
lysate on hindgut contractions of the enzootic vector,
L. longipalpis.
Herpetomonas muscarum lysate proteins (60 g/ml)
were tested on spontaneous contractions of hindgut and
oviduct in the natural host Musca domestica, but no
eVect was detected (results not shown). Lysate proteins
of C. fasciculata and H. muscarum were controls for
other insect hindgut, oviduct, and dorsal blood vessel
preparations. They did not inhibit muscle contractions
in any insect assay. These two protozoan species are
obligate gut parasites of mosquitoes and house Xies,
respectively. Both parasites are monoxenous; they exist
only in their insect hosts and are not transmitted to ver-
tebrates, unlike Leishmania spp. which alternate between
insect and vertebrate hosts. These parasites bind to the
insect gut and are not free in the lumen. Thus, they likely
employ mechanical adaptations such as Xagellar hemi-
desmosomes or interdigitating Xagella between micro-
villi (Brooker, 1971; Romeiro et al., 2003) to persist in
the insect gut.
Leptomonas seymouri was the only non-Leishmania
kinetoplastid in which myoinhibitory activity was
detected (»5%). L. seymouri is an obligate monoxenous
parasite of the cotton stainer bug Dysdercus suturellus
(Hemiptera) (Wallace, 1977). There is little information
on D. suturellus gut physiology and this protozoan, and
it is diYcult to speculate how myoinhibition of host gut
contractions may beneWt the parasites (Romeiro et al.,
2003).
Acknowledgments
I am grateful for the guidance and support of Y.
Schlein, J. Shlomai, and A. Warburg at the Kuvin Centre
for the Study of Infectious and Tropical Diseases, Had-
assah Medical School, Jerusalem. L. Schnur provided
seed cultures for all parasites. R.L. Jacobson helped in
culturing parasites.
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