Human toxoplasmosis is caused by the intracellular protozoan parasite Toxoplasma gondii. Although T. gondii infection is generally asymptomatic for most of the immunocompetent adults, severe complications may occur pa...Human toxoplasmosis is caused by the intracellular protozoan parasite Toxoplasma gondii. Although T. gondii infection is generally asymptomatic for most of the immunocompetent adults, severe complications may occur particularly in pregnant women and immunocompromised individual. Host cell immunity plays a critical role in parasite differentiation and persistence in the host. Therefore, genetic polymorphism in the host immune genes, for instance interferon-γ gene could be linked with possibility of T. gondii infection. The objective of the study was to verify the link between the single nucleotide polymorphisms (SNPs) in the IFN-γ gene of pregnant women and T. gondii infection through correlating with anthropometric and sociodemographic parameters. In this study, ninety-two (N = 92) pregnant women (16 - 40 years) and healthy controls (N = 95) with similar age ranges were included. Among them, 25% (n = 23) pregnant women were seropositive for T. gondii IgG antibodies by Rapid Test Assay. Allelic and genotypic frequencies of IFN-γ +874T/A (rs2430561) SNPs were evaluated by using ARMS-PCR. The distribution of the A and T alleles in the specific position of the IFN-γ gene in the T. gondii-infected pregnant women and the control groups did not differ significantly, according to the data. However, we found a higher frequency (13.04%) of A/A genotype in T. gondii infected pregnant women as compared to non-infected individuals (8.70%), demonstrating that T. gondii infection susceptibility may be increased by homozygosity for the A allele. Further studies are to be needed to find out the link between host gene polymorphism and T. gondii infection in Bangladesh.展开更多
Objective:To determine the detection rate of anti-Toxoplasma gondii(T.gondii) IgG and IgM in chronic HCV patients attending the Department of Tropical Medicine Mansoura University hospital in Egypt.Methods:This study ...Objective:To determine the detection rate of anti-Toxoplasma gondii(T.gondii) IgG and IgM in chronic HCV patients attending the Department of Tropical Medicine Mansoura University hospital in Egypt.Methods:This study included 120 adult chronic HCV patients.81 decompensate cirrhosis(late-stage)and 39 chronic HCV non cirrhotic patients(early-stage) and40 healthy blood donors as controls.Serum samples uere examined for anti-Toxoplasma IgM and anti-Toxoplasma IgG antibodies by ELISA.Real-time RT-polymerase chain reaction assay was done for quantitation of hepatitis C virus.Results:Anti-T.gondii IgG antibodies were detected in 75(92.6%) of 81 late-stage cirrhotic patients.30(76.9%) of the 39 chronic HCV non cirrhotic patients(early-Stage) and in 6(15ft) of 40 controls with statistically significant difference(P<0.001).Anti-T.gondii IgM antibodies were found in 11(13.6%) in late stage patients,5(12.8%)in early stage and in 3(7.5%) of controls with no statistical significant difference(P=0.610).There was no correlation between stage of fibrosis and IgM or IgG antibodies positivity in our studied groups(P=0.526).High IgG levels significantly correlated with high viral load(P=0.026).Conclusions:Our findings suggest that the serious opportunistic T.gondii infection represent a potential significant risk for chronic HCV patients.So.toxoplasmosis should be considered in their investigations and follow-up.展开更多
Objective: To know the difference between chemerin and adipocyte fatty acid-binding protein(AFABP) levels in obese individuals with positive Toxoplasma gondii(T. gondii)immunoglobulin G(IgG) compared with negative T. ...Objective: To know the difference between chemerin and adipocyte fatty acid-binding protein(AFABP) levels in obese individuals with positive Toxoplasma gondii(T. gondii)immunoglobulin G(IgG) compared with negative T. gondii IgG.Methods: This study is a cross-sectional study by using consecutive sampling methods conducted from January to April 2013. The subjects were 57 obese individuals who were divided into obese group of positive and negative T. gondii IgG. The level of chemerin,AFABP and T. gondii IgG was done by ELISA. The data were analyzed by independent t test.Results: The results showed that the level of chemerin of positive T. gondii IgG group was significantly higher than the negative T. gondii IgG group [(70.0 ± 16.5) vs.(64.4 ± 16.1) pg/mL; P = 0.003], but there was not significant AFABP difference between seropositive and negative IgG groups [(83.6 ± 41.9) vs.(74.2 ± 36.7) pg/mL; P = 0.598].Conclusions: It can be concluded that the level of chemerin of seropositive T. gondii IgG was higher than that in the negative T. gondii IgG group.展开更多
AIM:To detect whether Toxoplasma gondii(T.gondii)infection of mice can induce retinal DNA damage.METHODS:A total of 20 laboratory-bred male Swiss albino mice were used and divided into four groups:control group(non-in...AIM:To detect whether Toxoplasma gondii(T.gondii)infection of mice can induce retinal DNA damage.METHODS:A total of 20 laboratory-bred male Swiss albino mice were used and divided into four groups:control group(non-infected animals);T.gondii infected group;immunosuppressed infected group;and infected group treated with sulfadiazine and pyrimethamine.Mice eyes were collected 6wk post infection and retinas were obtained.Each retina was immediately processed for comet assay and the frequency of tailed nuclei(DNA damage)was calculated.In addition,retinal DNA damage was revealed by various comet assay parameters that were provided by the image analysis software including tail length,percentage of DNA in the tail,percentage of tailed cells and tail moment.RESULTS:The obtained results showed that T.gondii infection induced a statistically significant increase in the frequency of tailed nuclei,tail length,percentage of DNA in the tail,and tail moment in mice retinal cells compared to the control group(which showed some degree of DNA damage).In immunosuppressed infected group,retinal DNA damage was severing and there was significant increase in various comet assay parameters compared to both control and infected groups.After treatment with sulfadiazine and pyrimethamine,retinal DNA damage decreased and all comet assay parameters showed a statistical significant decrease compared to infected groups.CONCLUSION:T.gondii infection can induce DNA damage in mice retinal cells.展开更多
Background:Few investigations of genotype II of Toxoplasma gondii,the most preva-lent form of the Toxoplasma parasite in humans,have been carried out on due to the rapid conversion of tachyzoites to bradyzoites in its...Background:Few investigations of genotype II of Toxoplasma gondii,the most preva-lent form of the Toxoplasma parasite in humans,have been carried out on due to the rapid conversion of tachyzoites to bradyzoites in its life cycle.The current study aimed to create animal and in vitro models for production of the tachyzoites of the Prugniaud(PRU)genotype II strain.Methods:To develop an immunocompromised model and obtain tachyzoites of the PRU strain,BALB/c mice were orally treated with dexamethasone(10 mg/kg),cyclo-phosphamide(36 mg/kg),and cyclosporine(18 mg/kg)from 5 days prior to inocula-tion.Then,10-15 tissue cysts of PRU strain were inoculated intraperitoneally into the mice.The tachyzoites obtained from mice were then cultivated in a HeLa cell culture.The resulting yield of tachyzoites was cryopreserved in 92%fetal calf serum,8%dimethyl sulfoxide.The infectivity of these tachyzoites was evaluated using in vivo and in vitro examinations.Results:Numerous tachyzoites were observed in the peritoneal fluid of the immuno-suppressed mice within 10-15 days after inoculation,and many tachyzoites were har-vested from the HeLa cell culture.Trypan Blue staining showed 80%viability of the tachyzoites recovered from cryopreservation and this was confirmed by HeLa cell culture.In addition,mice infected intraperitoneally with the recovered tachyzoites presented with cysts in the brain after 2 months.Conclusion:We have developed an animal model for mass production of T.gondii tachyzoites of the PRU strain.This method can provide fresh viable tachyzoites of Toxoplasma gondii for use as and when required in future investigations.展开更多
Toxoplasma gondii (T. gondii) an intracellular protozoan parasite, infects mammals including human population world-wide. Upon primary infection, the parasite contributes to mild flu like symptoms in immune competent ...Toxoplasma gondii (T. gondii) an intracellular protozoan parasite, infects mammals including human population world-wide. Upon primary infection, the parasite contributes to mild flu like symptoms in immune competent host, but life threatening complication is seen in immune compromised patients and in pregnant women. Understanding the host-parasite interaction is critical for understanding the pathogenesis and biology parasite reactivation in the host. In this study, we used proteotrasncriptomics analyses by integrating the transcriptomics and proteomics data of T. gondii infected mouse liver to uncover the effector molecules responsible for disease pathogenesis that can be used as candidate markers for diagnosis and drug target. With this aim, we systematically integrated transcriptomicand proteomic data, representing the parasite infected mouse liver. Out of 2758 differentially expressed genes (DEGs) and 301 differentially expressed proteins (DEPs), 159 overlapping genes were identified. Among them, 86 genes were upregulated and 72 were downregulated in their respective mRNA and protein levels in the infected condition. Gene Ontology (GO) analysis revealed that the upregulated genes were mostly associated with immune system processes whereas the downregulated genes were involved in oxidation-reduction process and metabolism of lipid, and fatty acids. Protein-protein interaction (PPI) network analysis uncovered an interaction-hub including, Psmb8, Psmb9 and Tap1 for upregulated proteins and Cyp1A2, Cyp4A10 and Cyp3A11 for down-regulated proteins. Further studies are needed to validating these effector molecules. These molecules are likely to play a vital role in disease pathogenesis, as well as can be used as potential diagnostic marker and drug target candidates.展开更多
<i>Toxoplasma gondii</i> is a zoonotic protozoan that can infect any warm-blooded mammal. <i>T. gondii</i> infects about one-third of the human population on the planet. Infection with the para...<i>Toxoplasma gondii</i> is a zoonotic protozoan that can infect any warm-blooded mammal. <i>T. gondii</i> infects about one-third of the human population on the planet. Infection with the parasite in human causes toxoplasmosis that may pose a high risk in immunocompromised individuals under certain clinical conditions. Cats are the ultimate hosts of <i>T. gondii</i> where oocysts are formed through mating of male and female gametes. Infected cats can expel <i>T. gondii</i> oocysts in their feces, and thereby capable of pass on a disease to humans and other animals through consumption of foods, vegetables and water that are polluted with cat feces. The study was conducted to detect the presence of anti-<i>T. gondii</i> IgM and IgG antibodies in the blood of individuals with or without cat contact to determine if there is any relationship between cat contacts and <i>T. gondii</i> infection in humans. To address this, we enrolled subjects who contacted with the cat as target group and individuals with no cat contact as control group. Following register of different demographic data (including age, sex, education, foods habit, income status, etc.), whole blood from each enrolled subject of both the target group and control group was collected for serum preparation. <i>T. gondii</i> infected subjects were detected by Toxo Rapid test kit through identifying anti-<i>T. gondii</i> IgM and IgG antibodies in their serum. We found that only three out of twenty subjects who were in contact with cat showed positive IgG response while IgM antibody response was absent for all subjects. When compared with the data from control group, we did not find any significant association (p = 0.33) of cat contact with the transmission <i>of T. gondii</i> into human. However, with this small number of study subjects, we cannot conclusively say that there is no impact of cat contact on the transmission of <i>T. gondii</i> into human. Whether any association exists or not can be ascertained with a large number of subjects from different areas of Bangladesh in a future study in the population.展开更多
distribution.The main source of infection for humans is livestock and meat-producer animals.The relationships between Toxoplasma genotype and biological characteristics of the parasite have already been identified.Acc...distribution.The main source of infection for humans is livestock and meat-producer animals.The relationships between Toxoplasma genotype and biological characteristics of the parasite have already been identified.According to the pathogenicity of the parasite in laboratory animals,Toxoplasma is divided into three genotypes included type I,II and III.Understanding the genotype of the parasite,could help us to predict clinical features and severity of disease.The aim of this study was to identify genotypes of T.gondii in cattle and sheep meat and meat products in Ahvaz city southwest of Iran.One hundred and ninety samples of tongue,heart and muscles of sheep and cattle and meat products,including sausages and burgers,were collected from slaughterhouses and stores.To identify Toxoplasma gondii,DNA were extracted from samples and B1 gene were amplified by specific primers.To determine the genotype of T.gondii,PCR-RFLP was done on positive samples using by amplifying GRA6 gene and endonuclease Msel enzyme.Data analysis showed that the strain of the parasite in all positive samples belonged to genotype I.In this study the predominant Toxoplasma genotype was type I which can cause severe clinical symptoms in immunocompromised patients.Further research is needed to determine the genotype of the parasite in humans and other animals.展开更多
Background: Toxoplasma gondii (T. gondii) is an intracellular parasite mainly found in the central nervous system CNS, however, it can persist in multiple tissues in the body. Moreover, T. gondii is the commonest prot...Background: Toxoplasma gondii (T. gondii) is an intracellular parasite mainly found in the central nervous system CNS, however, it can persist in multiple tissues in the body. Moreover, T. gondii is the commonest protozoans causing infections among individuals with acquired immunodeficiency syndrome (AIDS). Thus, the aim of this study was to investigate the frequency of T. gondii infection among AIDS patients in Makkah at Saudi Arabia. Methods: Fifty patients with AIDS proved to be positive by ULTRA HIV Ag-Ab Enzyme Immunoassay, and thirty healthy volunteers negative for AIDS by ULTRA HIV Ag-Ab Enzyme Immunoassay were subjected to determination of anti T. gondii immunoglobulin M (IgM) antibody seropositivity and anti T. gondii immunoglobulin G (IgG) antibody seropositivity using commercially available enzyme-linked immunosorbent assay kits. Results: The results showed that the seropositivity rate of anti T. gondii IgM antibodies among AIDS patients (18%) was significantly higher than in the healthy volunteers group (3.33%). Regarding the serum level of anti T. gondii IgG antibodies among AIDS patients, it was 30% significantly higher compared with those of the seropositive healthy volunteers (6.67%). Conclusions: These statistically significant results support the association between T. gondii infection and AIDS and suggest the usefulness of providing data for an educational program that will be designed to prevent T. gondii infection among AIDS patients.展开更多
Objective:To compare analytical sensitivity and specificity of a newly described DNA amplification technique.LAMP and nested PCR assay targeting the RE and Bl genes for the detection of Toxoplasma gondii(T.gondii) DNA...Objective:To compare analytical sensitivity and specificity of a newly described DNA amplification technique.LAMP and nested PCR assay targeting the RE and Bl genes for the detection of Toxoplasma gondii(T.gondii) DNA.Methods:The analytical sensitivity of LAMP and ncstcd-PCR was obtained against 10-fold serial dilutions of T.gondii DNA ranging from 1 ng to 0.01 fg.DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays.Results:After testing LAMP and nesled-PCR in duplicate,the detection limit of RE-LAMP.B1-LAMP,RE-nested PCR and B1-nested PCR assays was one fg.100 fg,1 pg and 10 pg of T.gondii DNA respectively.All the LAMP assays and nested PCRs were 100% specific.The RE-LAMP assay revealed the most sensitivity for the detection of T.gondii DNA.Conclusions:The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T.gondii.Furthermore,these findings indicate that primers based on the RE are more suitable than those based on the B1 gene.However,the B1-LAMP assay has potential as a diagnostic tool for detection of T.gondii.展开更多
Objective: To identify serodiagnosis and quantification of Toxoplasma gondii(T. gondii) infection among pregnant women in Salmas, northwest of Iran. Methods: In this crosssectional study, 276 blood samples were collec...Objective: To identify serodiagnosis and quantification of Toxoplasma gondii(T. gondii) infection among pregnant women in Salmas, northwest of Iran. Methods: In this crosssectional study, 276 blood samples were collected from pregnant women referred to the health care centers in Salmas city. The demographic variables were also recorded. Titers of antiToxoplasma IgM and IgG antibodies(Ab) were determined using the chemiluminescence immunoassay. Quantitative real-time PCR targeting the T. gondii repeated element gene was also performed on the blood sample. Results: Out of all, 19.92%(55/276) and 2.17%(6/276) patients were seropositive for anti-Toxoplasma IgG and IgM Ab, respectively. Moreover, the presence of T. gondii DNA was observed in 12.31%(34/276) blood samples. A significant relationship was observed between the IgG Ab seropositivity and contact with the cat as a risk factor(P=0.022). Conclusions: The seroprevalence rate of T. gondii infection in pregnant women is relatively low. Consequently, the seronegative pregnant women are at risk, and a considerable rate of positive blood samples for the presence of parasite's DNA should not be ignored. Besides, quantitative real-time PCR could be considered as an accurate method for diagnosis of acute toxoplasmosis especially when the precise results are of the most importance in pregnancy. Limiting contact with cats is also suggested for pregnant women.展开更多
Objective:To evaluate parasite distribution and tissue tropism of Toxoplasma gondii tachyzoites in experimentally infected mice using real time QPCR.Methods:In this survey 16 Balb/c mice were inoculated with 1×10...Objective:To evaluate parasite distribution and tissue tropism of Toxoplasma gondii tachyzoites in experimentally infected mice using real time QPCR.Methods:In this survey 16 Balb/c mice were inoculated with 1×10~4 alive tachyzoites of Toxoplasma gondii RH strain.After 1,2,3 days post infection and the last day(before death),different tissues of mice including blood,brain,eye,liver,spleen,kidney,heart and muscle were harvested.Following tissues DNA extraction,the parasite burden was quantified using real time QPCR targeting the B1 gene(451 bp).Results:It showed that Toxoplasma after intraperitoneal injection was able to movement to various tissues in24 hours.Parasite burden was high in all tissues but the most number of parasites were observed in kidney,heart and liver,respectively.Conclusions:These data provide significant baseline information about Toxoplasma pathogenesis,vaccine monitoring and drug efficiency.展开更多
The objective of the present investigation was to estimate the prevalence of Toxoplasma gondii infection and co-infection with porcine reproductive and respiratory syndrome virus(PRRSV), classical swine fever virus(CS...The objective of the present investigation was to estimate the prevalence of Toxoplasma gondii infection and co-infection with porcine reproductive and respiratory syndrome virus(PRRSV), classical swine fever virus(CSFV) and porcine circovirus type 2(PCV-2) in pigs in China. A total of 372 tissues or serum samples collected from pigs distributed in 9 provinces/municipalities of China during the period from February 2011 to November 2012 were assayed for T. gondii antigens and antibodies using enzyme linked immunosorbent assay(ELISA) technique, while the PCR was designed for the detection of the PRRSV, CSFV and PCV-2, respectively. The total positive rate of T. gondii, PRSSV, CSFV and PCV-2 was 9.14%(34/372), 50.00%(186/372), 37.10%(138/372) and 3.23%(12/372), respectively. Among the 34 T. gondii positive samples, 26 samples were simultaneously infected with T. gondii and viruses, while the remaining eight samples were infected with T. gondii alone. In addition, the co-infection rate of T. gondii with PRSSV, T. gondii with PRSSV and CSFV, T. gondii with PRSSV and PCV-2, T. gondii with CSFV and PCV-2, T. gondii with PRSSV, CSFV and PCV-2 was 1.61%(6/372), 4.03%(15/372), 0.27%(1/372), 0.27%(1/372) and 0.81%(3/372), respectively. The results of the present survey revealed that PRRSV and CSFV were the common pathogens co-existing with porcine toxoplasmosis in China, and both of them could increase the chances of T. gondii infection in pig. This is the first report of T. gondii co-infections with viruses in pigs. It is very important to understand the interactions of parasite and virus, and can be used as reference data for the control and prevention of co-infections of T. gondii and viruses in pigs.展开更多
One strain of Toxoplasma gondii was successfully isolated from chickens in China by bioassay in mice. Antibodies and circulating antigens of T. gondii were assayed by the ELISA kits in 100 free range chickens from a r...One strain of Toxoplasma gondii was successfully isolated from chickens in China by bioassay in mice. Antibodies and circulating antigens of T. gondii were assayed by the ELISA kits in 100 free range chickens from a rural area surrounding Funing, China. Fifty-three chickens were antibody-positive and 21 chickens were antigen positive. Hearts, brains, spleens, lungs, livers, and kidneys of 21 antibody or antigen-positive chickens were bioassayed in mice. One strain of T. gondii was isolated from 1 of 21 (4.76%) chickens. The isolated T. gondii killed all of the inoculated mice. Genotyping of this isolate using polymorphisms at the loci 5′-SAG2, 3′-SAG2, SAG3, cB21-4, L358, BTUB, and GRA6 revealed that it was Type I. These indicated that it was virulent for mice. This is the first report of isolation of T. gondii from chickens in China.展开更多
Objective:To discuss the influence of tachyzoite of Toxoplasma gondii（T.gondii） RH strain on proliferation and apoptosis of hepatoma carcinoma（HCC） H7402 cell.Methods:The HCC H7402 cell in logarithmic phase and ta...Objective:To discuss the influence of tachyzoite of Toxoplasma gondii（T.gondii） RH strain on proliferation and apoptosis of hepatoma carcinoma（HCC） H7402 cell.Methods:The HCC H7402 cell in logarithmic phase and tachyzoite of T.gondii RH strain in different concentrations（1×107/mL,2×107/mL.4×107/mL,8×107mL and 16×107/mL） were co-cultured.CCK-8was utilized to determine the inhibition rate of T.gondii tachyzoite on H7402 cell growth.Flow cytometry was used to detect the change of cell cycle.RT-PCR method was used to detect the expression of cyclinB1 and cdc2-two genes related to cell cycle.Western blot method was used to detect the expression of apoptosis-related proteins Caspase-3 and Bcl-2.Results:The tachyzoite of T.gondii RH strain can inhibit the proliferation of HCC H7402 cells.The inhibition rate of tumor cell growth increased with the increase of concentration of T.gondii tachyzoite.With the increase of concentration of T.gondii tachyzoite,the proportion of G0/G1 phase of H7402 cell increased,the proportion of S phase decreased,and PI value decreased accordingly.The expression of cyclinB1 and cdc2 genes decreased with the increase of the concentration of T.gondii tachyzoite.With the increase of the concentration of tachyzoite of T.gondii RH strain,the expression quantity of Caspase-3 in H7402 cell increased,but the expression quantity of Bcl-2protein decreased.Conclusions:T.gondii can inhibit the in vitro proliferation of HCC H7402 cell,and induce its apoptosis.This effect shows a trend of concentration-dependent increase.Moreover,it is related to the down-regulation of cyclinB1 and cdc2（cell cycle-related genes）,the increase of apoptosis-related protein Caspase-3.and the decreasc of Bcl-2 expression.展开更多
Objective:To determine the seroprevalence of Toxoplasma gondii(T.gondii) infection in dogs and cats in Zhenjiang City,Jiangsu Province.Eastern China,and to evaluate the main associated risk factors relating to exposur...Objective:To determine the seroprevalence of Toxoplasma gondii(T.gondii) infection in dogs and cats in Zhenjiang City,Jiangsu Province.Eastern China,and to evaluate the main associated risk factors relating to exposure to 71 gondii in this region.Methods:Sera from 160 clogs and 116 cats from Zhenjiang City were tested for anti-T.gondii antibodies using EUSA.The seropositivity by area of activity,sex and age was analyzed.Results:Overall.21 dogs(13.l%) and 24 cats(20.7%) had antibodies to T.gondii.The infection rate in stray dogs(38.7%) and cats(28.6%! was significantly higher(P<0.05) than in household dogs(6.9%) and cats(18.2%).The seroprevalence in male clogs(14.8%) and cats(21.05%) were slighlly higher than their female counterparts(11.4%in dogs and 20.0%in cats),but were not significantly differenent(P>0.05).A high proportion of dogs at 3 to 6 years of age were positive to T.gondii(20.0%)while cats with relatively high seropositivity rates were at 0 to 1 year of age(33.3%).Conclusions:The prevalence of T.gondii infection in dogs and cats in Zhenjiang City was high,which is probably the main source of T.gondii infection in this area.展开更多
Objective The protozoan Toxoplasma gondii expresses large amounts of a 37 kDa Type 2C serine-threonine phosphatase,the so-called TgPP2 C which has been suggested to contribute to parasite growth regulation.Ectopic exp...Objective The protozoan Toxoplasma gondii expresses large amounts of a 37 kDa Type 2C serine-threonine phosphatase,the so-called TgPP2 C which has been suggested to contribute to parasite growth regulation.Ectopic expression in mammalian cells also indicated that the enzyme could regulate growth and survival.In this study,we aimed to investigate the interaction of TgPP2 C with human SSRP1(structure-specific recognition protein 1) and the effects of TgPP2 C on cell viability.Methods The yeast two hybrid system,His-tag pull-down and co-immunoprecipitation assays were used to confirm the interaction of TgPP2 C with SSRP1 and determine the binding domain on SSRP1.The evaluation of cell apoptosis was performed using cleaved caspase-3 antibody and Annexin-V/PI kit combined with flow cytometry.Results We identified human SSRP1 as an interacting partner of TgPP2 C.The C-terminal region of SSRP1 including the amino acids 471 to 538 was specifically mapped as the region responsible for interaction with TgPP2 C.The overexpression of TgPP2 C down-regulated cell apoptosis and negatively regulated apoptosis induced by DRB,casein kinase II(CKII) inhibitor,through enhanced interaction with SSRP1.Conclusion TgPP2 C may be a parasitic factor capable of promoting cell survival through interaction with the host protein SSRP1,thereby creating a favorable environment for parasite growth.展开更多
文摘Human toxoplasmosis is caused by the intracellular protozoan parasite Toxoplasma gondii. Although T. gondii infection is generally asymptomatic for most of the immunocompetent adults, severe complications may occur particularly in pregnant women and immunocompromised individual. Host cell immunity plays a critical role in parasite differentiation and persistence in the host. Therefore, genetic polymorphism in the host immune genes, for instance interferon-γ gene could be linked with possibility of T. gondii infection. The objective of the study was to verify the link between the single nucleotide polymorphisms (SNPs) in the IFN-γ gene of pregnant women and T. gondii infection through correlating with anthropometric and sociodemographic parameters. In this study, ninety-two (N = 92) pregnant women (16 - 40 years) and healthy controls (N = 95) with similar age ranges were included. Among them, 25% (n = 23) pregnant women were seropositive for T. gondii IgG antibodies by Rapid Test Assay. Allelic and genotypic frequencies of IFN-γ +874T/A (rs2430561) SNPs were evaluated by using ARMS-PCR. The distribution of the A and T alleles in the specific position of the IFN-γ gene in the T. gondii-infected pregnant women and the control groups did not differ significantly, according to the data. However, we found a higher frequency (13.04%) of A/A genotype in T. gondii infected pregnant women as compared to non-infected individuals (8.70%), demonstrating that T. gondii infection susceptibility may be increased by homozygosity for the A allele. Further studies are to be needed to find out the link between host gene polymorphism and T. gondii infection in Bangladesh.
文摘Objective:To determine the detection rate of anti-Toxoplasma gondii(T.gondii) IgG and IgM in chronic HCV patients attending the Department of Tropical Medicine Mansoura University hospital in Egypt.Methods:This study included 120 adult chronic HCV patients.81 decompensate cirrhosis(late-stage)and 39 chronic HCV non cirrhotic patients(early-stage) and40 healthy blood donors as controls.Serum samples uere examined for anti-Toxoplasma IgM and anti-Toxoplasma IgG antibodies by ELISA.Real-time RT-polymerase chain reaction assay was done for quantitation of hepatitis C virus.Results:Anti-T.gondii IgG antibodies were detected in 75(92.6%) of 81 late-stage cirrhotic patients.30(76.9%) of the 39 chronic HCV non cirrhotic patients(early-Stage) and in 6(15ft) of 40 controls with statistically significant difference(P<0.001).Anti-T.gondii IgM antibodies were found in 11(13.6%) in late stage patients,5(12.8%)in early stage and in 3(7.5%) of controls with no statistical significant difference(P=0.610).There was no correlation between stage of fibrosis and IgM or IgG antibodies positivity in our studied groups(P=0.526).High IgG levels significantly correlated with high viral load(P=0.026).Conclusions:Our findings suggest that the serious opportunistic T.gondii infection represent a potential significant risk for chronic HCV patients.So.toxoplasmosis should be considered in their investigations and follow-up.
基金Supported by Dean of Medical Faculty of Universitas Brawijaya and Direktorat Jendral of Higher Education,Ministry of National Education and Culture of Republic Indonesia with grant number of 0636/023-04.2.16/15/2012
文摘Objective: To know the difference between chemerin and adipocyte fatty acid-binding protein(AFABP) levels in obese individuals with positive Toxoplasma gondii(T. gondii)immunoglobulin G(IgG) compared with negative T. gondii IgG.Methods: This study is a cross-sectional study by using consecutive sampling methods conducted from January to April 2013. The subjects were 57 obese individuals who were divided into obese group of positive and negative T. gondii IgG. The level of chemerin,AFABP and T. gondii IgG was done by ELISA. The data were analyzed by independent t test.Results: The results showed that the level of chemerin of positive T. gondii IgG group was significantly higher than the negative T. gondii IgG group [(70.0 ± 16.5) vs.(64.4 ± 16.1) pg/mL; P = 0.003], but there was not significant AFABP difference between seropositive and negative IgG groups [(83.6 ± 41.9) vs.(74.2 ± 36.7) pg/mL; P = 0.598].Conclusions: It can be concluded that the level of chemerin of seropositive T. gondii IgG was higher than that in the negative T. gondii IgG group.
文摘AIM:To detect whether Toxoplasma gondii(T.gondii)infection of mice can induce retinal DNA damage.METHODS:A total of 20 laboratory-bred male Swiss albino mice were used and divided into four groups:control group(non-infected animals);T.gondii infected group;immunosuppressed infected group;and infected group treated with sulfadiazine and pyrimethamine.Mice eyes were collected 6wk post infection and retinas were obtained.Each retina was immediately processed for comet assay and the frequency of tailed nuclei(DNA damage)was calculated.In addition,retinal DNA damage was revealed by various comet assay parameters that were provided by the image analysis software including tail length,percentage of DNA in the tail,percentage of tailed cells and tail moment.RESULTS:The obtained results showed that T.gondii infection induced a statistically significant increase in the frequency of tailed nuclei,tail length,percentage of DNA in the tail,and tail moment in mice retinal cells compared to the control group(which showed some degree of DNA damage).In immunosuppressed infected group,retinal DNA damage was severing and there was significant increase in various comet assay parameters compared to both control and infected groups.After treatment with sulfadiazine and pyrimethamine,retinal DNA damage decreased and all comet assay parameters showed a statistical significant decrease compared to infected groups.CONCLUSION:T.gondii infection can induce DNA damage in mice retinal cells.
基金This research was funded by the project 97-01-01-18897 from Shiraz University of Medical Sciences,Shiraz,Iran.
文摘Background:Few investigations of genotype II of Toxoplasma gondii,the most preva-lent form of the Toxoplasma parasite in humans,have been carried out on due to the rapid conversion of tachyzoites to bradyzoites in its life cycle.The current study aimed to create animal and in vitro models for production of the tachyzoites of the Prugniaud(PRU)genotype II strain.Methods:To develop an immunocompromised model and obtain tachyzoites of the PRU strain,BALB/c mice were orally treated with dexamethasone(10 mg/kg),cyclo-phosphamide(36 mg/kg),and cyclosporine(18 mg/kg)from 5 days prior to inocula-tion.Then,10-15 tissue cysts of PRU strain were inoculated intraperitoneally into the mice.The tachyzoites obtained from mice were then cultivated in a HeLa cell culture.The resulting yield of tachyzoites was cryopreserved in 92%fetal calf serum,8%dimethyl sulfoxide.The infectivity of these tachyzoites was evaluated using in vivo and in vitro examinations.Results:Numerous tachyzoites were observed in the peritoneal fluid of the immuno-suppressed mice within 10-15 days after inoculation,and many tachyzoites were har-vested from the HeLa cell culture.Trypan Blue staining showed 80%viability of the tachyzoites recovered from cryopreservation and this was confirmed by HeLa cell culture.In addition,mice infected intraperitoneally with the recovered tachyzoites presented with cysts in the brain after 2 months.Conclusion:We have developed an animal model for mass production of T.gondii tachyzoites of the PRU strain.This method can provide fresh viable tachyzoites of Toxoplasma gondii for use as and when required in future investigations.
文摘Toxoplasma gondii (T. gondii) an intracellular protozoan parasite, infects mammals including human population world-wide. Upon primary infection, the parasite contributes to mild flu like symptoms in immune competent host, but life threatening complication is seen in immune compromised patients and in pregnant women. Understanding the host-parasite interaction is critical for understanding the pathogenesis and biology parasite reactivation in the host. In this study, we used proteotrasncriptomics analyses by integrating the transcriptomics and proteomics data of T. gondii infected mouse liver to uncover the effector molecules responsible for disease pathogenesis that can be used as candidate markers for diagnosis and drug target. With this aim, we systematically integrated transcriptomicand proteomic data, representing the parasite infected mouse liver. Out of 2758 differentially expressed genes (DEGs) and 301 differentially expressed proteins (DEPs), 159 overlapping genes were identified. Among them, 86 genes were upregulated and 72 were downregulated in their respective mRNA and protein levels in the infected condition. Gene Ontology (GO) analysis revealed that the upregulated genes were mostly associated with immune system processes whereas the downregulated genes were involved in oxidation-reduction process and metabolism of lipid, and fatty acids. Protein-protein interaction (PPI) network analysis uncovered an interaction-hub including, Psmb8, Psmb9 and Tap1 for upregulated proteins and Cyp1A2, Cyp4A10 and Cyp3A11 for down-regulated proteins. Further studies are needed to validating these effector molecules. These molecules are likely to play a vital role in disease pathogenesis, as well as can be used as potential diagnostic marker and drug target candidates.
文摘<i>Toxoplasma gondii</i> is a zoonotic protozoan that can infect any warm-blooded mammal. <i>T. gondii</i> infects about one-third of the human population on the planet. Infection with the parasite in human causes toxoplasmosis that may pose a high risk in immunocompromised individuals under certain clinical conditions. Cats are the ultimate hosts of <i>T. gondii</i> where oocysts are formed through mating of male and female gametes. Infected cats can expel <i>T. gondii</i> oocysts in their feces, and thereby capable of pass on a disease to humans and other animals through consumption of foods, vegetables and water that are polluted with cat feces. The study was conducted to detect the presence of anti-<i>T. gondii</i> IgM and IgG antibodies in the blood of individuals with or without cat contact to determine if there is any relationship between cat contacts and <i>T. gondii</i> infection in humans. To address this, we enrolled subjects who contacted with the cat as target group and individuals with no cat contact as control group. Following register of different demographic data (including age, sex, education, foods habit, income status, etc.), whole blood from each enrolled subject of both the target group and control group was collected for serum preparation. <i>T. gondii</i> infected subjects were detected by Toxo Rapid test kit through identifying anti-<i>T. gondii</i> IgM and IgG antibodies in their serum. We found that only three out of twenty subjects who were in contact with cat showed positive IgG response while IgM antibody response was absent for all subjects. When compared with the data from control group, we did not find any significant association (p = 0.33) of cat contact with the transmission <i>of T. gondii</i> into human. However, with this small number of study subjects, we cannot conclusively say that there is no impact of cat contact on the transmission of <i>T. gondii</i> into human. Whether any association exists or not can be ascertained with a large number of subjects from different areas of Bangladesh in a future study in the population.
基金This study financially supported by Ahvaz Jundishapur University of Medical Sciences with number grant:90111.
文摘distribution.The main source of infection for humans is livestock and meat-producer animals.The relationships between Toxoplasma genotype and biological characteristics of the parasite have already been identified.According to the pathogenicity of the parasite in laboratory animals,Toxoplasma is divided into three genotypes included type I,II and III.Understanding the genotype of the parasite,could help us to predict clinical features and severity of disease.The aim of this study was to identify genotypes of T.gondii in cattle and sheep meat and meat products in Ahvaz city southwest of Iran.One hundred and ninety samples of tongue,heart and muscles of sheep and cattle and meat products,including sausages and burgers,were collected from slaughterhouses and stores.To identify Toxoplasma gondii,DNA were extracted from samples and B1 gene were amplified by specific primers.To determine the genotype of T.gondii,PCR-RFLP was done on positive samples using by amplifying GRA6 gene and endonuclease Msel enzyme.Data analysis showed that the strain of the parasite in all positive samples belonged to genotype I.In this study the predominant Toxoplasma genotype was type I which can cause severe clinical symptoms in immunocompromised patients.Further research is needed to determine the genotype of the parasite in humans and other animals.
文摘Background: Toxoplasma gondii (T. gondii) is an intracellular parasite mainly found in the central nervous system CNS, however, it can persist in multiple tissues in the body. Moreover, T. gondii is the commonest protozoans causing infections among individuals with acquired immunodeficiency syndrome (AIDS). Thus, the aim of this study was to investigate the frequency of T. gondii infection among AIDS patients in Makkah at Saudi Arabia. Methods: Fifty patients with AIDS proved to be positive by ULTRA HIV Ag-Ab Enzyme Immunoassay, and thirty healthy volunteers negative for AIDS by ULTRA HIV Ag-Ab Enzyme Immunoassay were subjected to determination of anti T. gondii immunoglobulin M (IgM) antibody seropositivity and anti T. gondii immunoglobulin G (IgG) antibody seropositivity using commercially available enzyme-linked immunosorbent assay kits. Results: The results showed that the seropositivity rate of anti T. gondii IgM antibodies among AIDS patients (18%) was significantly higher than in the healthy volunteers group (3.33%). Regarding the serum level of anti T. gondii IgG antibodies among AIDS patients, it was 30% significantly higher compared with those of the seropositive healthy volunteers (6.67%). Conclusions: These statistically significant results support the association between T. gondii infection and AIDS and suggest the usefulness of providing data for an educational program that will be designed to prevent T. gondii infection among AIDS patients.
基金supported financially by grant of Lorestan University of Medical Sciences,Khorramabad,Iran
文摘Objective:To compare analytical sensitivity and specificity of a newly described DNA amplification technique.LAMP and nested PCR assay targeting the RE and Bl genes for the detection of Toxoplasma gondii(T.gondii) DNA.Methods:The analytical sensitivity of LAMP and ncstcd-PCR was obtained against 10-fold serial dilutions of T.gondii DNA ranging from 1 ng to 0.01 fg.DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays.Results:After testing LAMP and nesled-PCR in duplicate,the detection limit of RE-LAMP.B1-LAMP,RE-nested PCR and B1-nested PCR assays was one fg.100 fg,1 pg and 10 pg of T.gondii DNA respectively.All the LAMP assays and nested PCRs were 100% specific.The RE-LAMP assay revealed the most sensitivity for the detection of T.gondii DNA.Conclusions:The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T.gondii.Furthermore,these findings indicate that primers based on the RE are more suitable than those based on the B1 gene.However,the B1-LAMP assay has potential as a diagnostic tool for detection of T.gondii.
基金supported by Infectious and Tropical Disease Research Center,Tabriz University of Medical Sciences,Tabriz,Iran(Grant No.94/2-5/17)
文摘Objective: To identify serodiagnosis and quantification of Toxoplasma gondii(T. gondii) infection among pregnant women in Salmas, northwest of Iran. Methods: In this crosssectional study, 276 blood samples were collected from pregnant women referred to the health care centers in Salmas city. The demographic variables were also recorded. Titers of antiToxoplasma IgM and IgG antibodies(Ab) were determined using the chemiluminescence immunoassay. Quantitative real-time PCR targeting the T. gondii repeated element gene was also performed on the blood sample. Results: Out of all, 19.92%(55/276) and 2.17%(6/276) patients were seropositive for anti-Toxoplasma IgG and IgM Ab, respectively. Moreover, the presence of T. gondii DNA was observed in 12.31%(34/276) blood samples. A significant relationship was observed between the IgG Ab seropositivity and contact with the cat as a risk factor(P=0.022). Conclusions: The seroprevalence rate of T. gondii infection in pregnant women is relatively low. Consequently, the seronegative pregnant women are at risk, and a considerable rate of positive blood samples for the presence of parasite's DNA should not be ignored. Besides, quantitative real-time PCR could be considered as an accurate method for diagnosis of acute toxoplasmosis especially when the precise results are of the most importance in pregnancy. Limiting contact with cats is also suggested for pregnant women.
基金prepared from Yousef Dadimoghaddam's MScthesis and supported by grant(no.90-31) from Deputy of Research,Mazandaran University of Medical Sciences.Sari,IranThe spousor or Junding organization had norole in the design or conduct of this research
文摘Objective:To evaluate parasite distribution and tissue tropism of Toxoplasma gondii tachyzoites in experimentally infected mice using real time QPCR.Methods:In this survey 16 Balb/c mice were inoculated with 1×10~4 alive tachyzoites of Toxoplasma gondii RH strain.After 1,2,3 days post infection and the last day(before death),different tissues of mice including blood,brain,eye,liver,spleen,kidney,heart and muscle were harvested.Following tissues DNA extraction,the parasite burden was quantified using real time QPCR targeting the B1 gene(451 bp).Results:It showed that Toxoplasma after intraperitoneal injection was able to movement to various tissues in24 hours.Parasite burden was high in all tissues but the most number of parasites were observed in kidney,heart and liver,respectively.Conclusions:These data provide significant baseline information about Toxoplasma pathogenesis,vaccine monitoring and drug efficiency.
基金supported by the Special Fund for Public Welfare Industry of Ministry of Agriculture of China (20090303604)the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (PAPD)
文摘The objective of the present investigation was to estimate the prevalence of Toxoplasma gondii infection and co-infection with porcine reproductive and respiratory syndrome virus(PRRSV), classical swine fever virus(CSFV) and porcine circovirus type 2(PCV-2) in pigs in China. A total of 372 tissues or serum samples collected from pigs distributed in 9 provinces/municipalities of China during the period from February 2011 to November 2012 were assayed for T. gondii antigens and antibodies using enzyme linked immunosorbent assay(ELISA) technique, while the PCR was designed for the detection of the PRRSV, CSFV and PCV-2, respectively. The total positive rate of T. gondii, PRSSV, CSFV and PCV-2 was 9.14%(34/372), 50.00%(186/372), 37.10%(138/372) and 3.23%(12/372), respectively. Among the 34 T. gondii positive samples, 26 samples were simultaneously infected with T. gondii and viruses, while the remaining eight samples were infected with T. gondii alone. In addition, the co-infection rate of T. gondii with PRSSV, T. gondii with PRSSV and CSFV, T. gondii with PRSSV and PCV-2, T. gondii with CSFV and PCV-2, T. gondii with PRSSV, CSFV and PCV-2 was 1.61%(6/372), 4.03%(15/372), 0.27%(1/372), 0.27%(1/372) and 0.81%(3/372), respectively. The results of the present survey revealed that PRRSV and CSFV were the common pathogens co-existing with porcine toxoplasmosis in China, and both of them could increase the chances of T. gondii infection in pig. This is the first report of T. gondii co-infections with viruses in pigs. It is very important to understand the interactions of parasite and virus, and can be used as reference data for the control and prevention of co-infections of T. gondii and viruses in pigs.
基金supported by the Special Fund for Public Welfare Industry of Chinese Ministry of Agriculture(200903036-04)
文摘One strain of Toxoplasma gondii was successfully isolated from chickens in China by bioassay in mice. Antibodies and circulating antigens of T. gondii were assayed by the ELISA kits in 100 free range chickens from a rural area surrounding Funing, China. Fifty-three chickens were antibody-positive and 21 chickens were antigen positive. Hearts, brains, spleens, lungs, livers, and kidneys of 21 antibody or antigen-positive chickens were bioassayed in mice. One strain of T. gondii was isolated from 1 of 21 (4.76%) chickens. The isolated T. gondii killed all of the inoculated mice. Genotyping of this isolate using polymorphisms at the loci 5′-SAG2, 3′-SAG2, SAG3, cB21-4, L358, BTUB, and GRA6 revealed that it was Type I. These indicated that it was virulent for mice. This is the first report of isolation of T. gondii from chickens in China.
文摘Objective:To discuss the influence of tachyzoite of Toxoplasma gondii（T.gondii） RH strain on proliferation and apoptosis of hepatoma carcinoma（HCC） H7402 cell.Methods:The HCC H7402 cell in logarithmic phase and tachyzoite of T.gondii RH strain in different concentrations（1×107/mL,2×107/mL.4×107/mL,8×107mL and 16×107/mL） were co-cultured.CCK-8was utilized to determine the inhibition rate of T.gondii tachyzoite on H7402 cell growth.Flow cytometry was used to detect the change of cell cycle.RT-PCR method was used to detect the expression of cyclinB1 and cdc2-two genes related to cell cycle.Western blot method was used to detect the expression of apoptosis-related proteins Caspase-3 and Bcl-2.Results:The tachyzoite of T.gondii RH strain can inhibit the proliferation of HCC H7402 cells.The inhibition rate of tumor cell growth increased with the increase of concentration of T.gondii tachyzoite.With the increase of concentration of T.gondii tachyzoite,the proportion of G0/G1 phase of H7402 cell increased,the proportion of S phase decreased,and PI value decreased accordingly.The expression of cyclinB1 and cdc2 genes decreased with the increase of the concentration of T.gondii tachyzoite.With the increase of the concentration of tachyzoite of T.gondii RH strain,the expression quantity of Caspase-3 in H7402 cell increased,but the expression quantity of Bcl-2protein decreased.Conclusions:T.gondii can inhibit the in vitro proliferation of HCC H7402 cell,and induce its apoptosis.This effect shows a trend of concentration-dependent increase.Moreover,it is related to the down-regulation of cyclinB1 and cdc2（cell cycle-related genes）,the increase of apoptosis-related protein Caspase-3.and the decreasc of Bcl-2 expression.
基金Supported by the Project of Access Engineers of Higher Vocational Institutions in Jiangsu Province(Project No.2013FG042)Jiangsu Polytechnic College of Agriculture and Forestry
文摘Objective:To determine the seroprevalence of Toxoplasma gondii(T.gondii) infection in dogs and cats in Zhenjiang City,Jiangsu Province.Eastern China,and to evaluate the main associated risk factors relating to exposure to 71 gondii in this region.Methods:Sera from 160 clogs and 116 cats from Zhenjiang City were tested for anti-T.gondii antibodies using EUSA.The seropositivity by area of activity,sex and age was analyzed.Results:Overall.21 dogs(13.l%) and 24 cats(20.7%) had antibodies to T.gondii.The infection rate in stray dogs(38.7%) and cats(28.6%! was significantly higher(P<0.05) than in household dogs(6.9%) and cats(18.2%).The seroprevalence in male clogs(14.8%) and cats(21.05%) were slighlly higher than their female counterparts(11.4%in dogs and 20.0%in cats),but were not significantly differenent(P>0.05).A high proportion of dogs at 3 to 6 years of age were positive to T.gondii(20.0%)while cats with relatively high seropositivity rates were at 0 to 1 year of age(33.3%).Conclusions:The prevalence of T.gondii infection in dogs and cats in Zhenjiang City was high,which is probably the main source of T.gondii infection in this area.
基金supported by the Natural Science Foundation of Guangdong Province(Grant No.9151065004000005)National Program on Key Basic Research Project(973 Program)(Grant No.2011CB910700)+6 种基金High-Level Talents Project of the Universities of Guangdong(No.[2011]431)National Natural Science Foundation of China(Grant No.3100062881071790)Natural Science Foundation of Guangdong Province(Grant No.S2013030013315)Fundamental Research Funds for the Central Universities(Grant No.216114302161010121609317)
文摘Objective The protozoan Toxoplasma gondii expresses large amounts of a 37 kDa Type 2C serine-threonine phosphatase,the so-called TgPP2 C which has been suggested to contribute to parasite growth regulation.Ectopic expression in mammalian cells also indicated that the enzyme could regulate growth and survival.In this study,we aimed to investigate the interaction of TgPP2 C with human SSRP1(structure-specific recognition protein 1) and the effects of TgPP2 C on cell viability.Methods The yeast two hybrid system,His-tag pull-down and co-immunoprecipitation assays were used to confirm the interaction of TgPP2 C with SSRP1 and determine the binding domain on SSRP1.The evaluation of cell apoptosis was performed using cleaved caspase-3 antibody and Annexin-V/PI kit combined with flow cytometry.Results We identified human SSRP1 as an interacting partner of TgPP2 C.The C-terminal region of SSRP1 including the amino acids 471 to 538 was specifically mapped as the region responsible for interaction with TgPP2 C.The overexpression of TgPP2 C down-regulated cell apoptosis and negatively regulated apoptosis induced by DRB,casein kinase II(CKII) inhibitor,through enhanced interaction with SSRP1.Conclusion TgPP2 C may be a parasitic factor capable of promoting cell survival through interaction with the host protein SSRP1,thereby creating a favorable environment for parasite growth.
