The complete mitochondrial genomes for three Toxocara species of human and animal health significance

<p>Abstract</p> <p>Background</p> <p>Studying mitochondrial (mt) genomics has important implications for various fundamental areas, including mt biochemistry, physiology and molecular biology. In addition, mt genome sequences have provided useful markers for investigating population genetic structures, systematics and phylogenetics of organisms. <it>Toxocara canis, Toxocara cati </it>and <it>Toxocara malaysiensis </it>cause significant health problems in animals and humans. Although they are of importance in human and animal health, no information on the mt genomes for any of <it>Toxocara </it>species is available.</p> <p>Results</p> <p>The sizes of the entire mt genome are 14,322 bp for <it>T. canis</it>, 14029 bp for <it>T. cati </it>and 14266 bp for <it>T. malaysiensis</it>, respectively. These circular genomes are amongst the largest reported to date for all secernentean nematodes. Their relatively large sizes relate mainly to an increased length in the AT-rich region. The mt genomes of the three <it>Toxocara </it>species all encode 12 proteins, two ribosomal RNAs and 22 transfer RNA genes, but lack the ATP synthetase subunit 8 gene, which is consistent with all other species of Nematode studied to date, with the exception of <it>Trichinella spiralis</it>. All genes are transcribed in the same direction and have a nucleotide composition high in A and T, but low in G and C. The contents of A+T of the complete genomes are 68.57% for <it>T. canis</it>, 69.95% for <it>T. cati </it>and 68.86% for <it>T. malaysiensis</it>, among which the A+T for <it>T. canis </it>is the lowest among all nematodes studied to date. The AT bias had a significant effect on both the codon usage pattern and amino acid composition of proteins. The mt genome structures for three <it>Toxocara </it>species, including genes and non-coding regions, are in the same order as for <it>Ascaris suum </it>and <it>Anisakis simplex</it>, but differ from <it>Ancylostoma duodenale</it>, <it>Necator americanus </it>and <it>Caenorhabditis elegans </it>only in the location of the AT-rich region, whereas there are substantial differences when compared with <it>Onchocerca volvulus</it>,<it>Dirofiliria immitis </it>and <it>Strongyloides stercoralis</it>. Phylogenetic analyses based on concatenated amino acid sequences of 12 protein-coding genes revealed that the newly described species <it>T. malaysiensis </it>was more closely related to <it>T. cati </it>than to <it>T. canis</it>, consistent with results of a previous study using sequences of nuclear internal transcribed spacers as genetic markers.</p> <p>Conclusion</p> <p>The present study determined the complete mt genome sequences for three roundworms of human and animal health significance, which provides mtDNA evidence for the validity of <it>T. malaysiensis </it>and also provides a foundation for studying the systematics, population genetics and ecology of these and other nematodes of socio-economic importance.</p

Changes in the proteomic profiles of mouse brain after infection with cyst-forming Toxoplasma gondii

Background Toxoplasma gondii is an opportunistic pathogenic protozoan parasite, which infects approximately one third of the human population worldwide, causing opportunistic zoonotic toxoplasmosis. The predilection of T. gondii for the central nervous system (CNS) causes behavioral disorders and fatal necrotizing encephalitis and thus constitutes a major threat especially to AIDS patients. Methods In the present study, we explored the proteomic profiles of brain tissues of the specific pathogen-free (SPF) Kunming mice at 7 d, 14 d and 21 d after infection with cysts of the Toxoplasma gondii Prugniaud (PRU) strain (Genotype II), by two-dimensional gel electrophoresis (2-DE) combined with MALDI-TOF/TOF tandem mass spectrometry (MS/MS). Results A total of 60 differentially expressed protein spots were selected. Fifty-six spots were successfully identified, which corresponded to 45 proteins of the mouse. Functional analysis using a Gene Ontology database showed that these proteins were mainly involved in metabolism, cell structure, signal transduction and immune responses, and will be beneficial for the understanding of molecular mechanisms of T. gondii pathogenesis. Conclusions This study identified some mouse brain proteins involved in the response with cyst-forming T. gondii PRU strain. These results provided an insight into the responsive relationship between T. gondii and the host brain tissues, which will shed light on our understanding of the mechanisms of pathogenesis in toxoplasmic encephalitis, and facilitate the discovery of new methods of diagnosis, prevention, control and treatment of toxoplasmic encephalopathy

Seroprevalence of Toxoplasma gondii infection in household and stray cats in Lanzhou, northwest China

<p>Abstract</p> <p>Background</p> <p><it>Toxoplasma gondii </it>is an important protozoan parasite infecting humans and almost all warm-blooded animals. As the only definitive host, cats play a crucial role in the transmission of <it>T. gondii </it>infection by shedding parasite oocysts in their feces. However, little information on <it>T. gondii </it>infection in cats was available in Lanzhou, northwest China. This study was performed to determine the seroprevalence of <it>T. gondii </it>infection in household and stray cats in Lanzhou, northwest China.</p> <p>Results</p> <p>A total of 221 (179 households and 42 strays) blood samples were collected from clinically healthy cats admitted to several pet hospitals located in Lanzhou City, between November 2010 and July 2011 for the serological detection of <it>T. gondii </it>infection. The majority (207) of these cats represented Chinese Lihua cats. 47 of 221 (21.3%) examined cats were seropositive for <it>T. gondii </it>infection using the modified agglutination test (MAT) at the cut-off of 1:25. The seroprevalence in household and stray cats was assessed to be 15.6% and 45.2%, respectively, and the difference was statistically significant (<it>P <</it>0.05). The seroprevalence ranged from 15.1% to 25.8% among different age groups, but the differences were not statistically significant (<it>P ></it>0.05). Studies showed that there was no relationship between seroprevalence and the gender (<it>P ></it>0.05).</p> <p>Conclusions</p> <p>The present survey indicated the high seroprevalence of <it>T. gondii </it>in cats in Lanzhou, northwest China, which poses a threat to animal and human health. Therefore, measures should be taken to control and prevent toxoplasmosis of cats in this area.</p

Withaferin A induces apoptosis in rat C6 glioma cells through regulating NF-ΚB nuclear translocation and activation of caspase cascade

Background: The demand for the chemopreventive drug from the plant source is increasing in recent times, owing to its various biological activities without any adverse effect. The intention of this current study was to examine the antiglioma effect of Withaferin A (WFA) on C6 glioma cell line model.Materials and Methods: C6 glioma cells were administrated with different concentration of WFA (50, 100, 200 and 500 μg/mL) and DMSO (control) group to examine its anti-proliferative, anti-inflammatory and pro-apoptotic activities.Results: Treatment with WFA showed a significant decline in the glioma cell count in a dose-dependent manner and thus proving its anti-proliferative effect. Similarly, inflammatory markers were also substantially lowered upon treatment with different concentration of WFA. However, DNA fragmentation and apoptotic markers like Caspase-3 and 9 were concomitantly enhanced after co-cultured with different concentration of WFA and thus exhibiting its cytotoxicity efficacy. Furthermore, the protein expression of Bcl2 and Bax were markedly downregulated and upregulated respectively; upon treatment with WFA on C6 glioma cells.Conclusion: The outcome of this study evidently demonstrates that C6 glioma cells co-cultured with increased concentration of WFA, showed an anti-proliferative, anti-inflammatory and pro-apoptotic effect in a dose-dependent fashion.Keywords: Withaferin A, C6 glioma cells, anti-proliferative, anti-inflammatory, caspase, apoptosi

Seroprevalence and Genetic Characterization of Toxoplasma Gondii in Three Species of Pet Birds in China

Background Toxoplasmosis, caused by the protozoan parasite Toxoplasma gondii, is one of the most common zoonosis worldwide, affecting a wide range of warm-blooded mammals and birds worldwide. However, no information on T. gondii infection in pet birds in China is available. Therefore, this study was performed to determine the prevalence of T. gondii infection in pet birds in Gansu province, China. Methods A total of 687 blood samples were collected from pet birds (Carduelis spinus, Alauda gulgula, Cocothraustes migratorlus) in three representative administrative regions in Gansu province, northwest China between August 2011 and September 2012 T. gondii antibodies were determined using the modified agglutination test (MAT). Genomic DNA was extracted from the brain tissues of seropositive pet birds and T. gondii B1 gene was amplified using a semi-nested PCR.DNA samples giving positive B1 amplification were then genetically characterized using multi-locus PCR-RFLP. Results The overall T. gondii seroprevalence was 11.21% (77/687). C. spinus had the highest T. gondii seroprevalence (11.65%), followed by A. arvensis (11.39%) and C. migratorlus (5.26%), these differences were not statistically significant (P \u3e 0.05). Of 77 DNA samples, 8 were positive for the T. gondii B1 gene, four showed complete genotyping results. Only one genotype (the Type II variant: ToxoDB genotype #3) was identified. Conclusions The results of the present survey indicated the presence of T. gondii infection in pet birds in Gansu province, China. These data provide base-line information for the execution of control strategies against T. gondii infection in pet birds. To our knowledge, this is the first report documenting the occurrence of T. gondii prevalence and genotype in pet birds in China

Prevalence of Sarcoptes scabiei

Little is known about the prevalence of Sarcoptes scabiei infection in pet dogs in China. In the present study, the prevalence of S. scabiei infection in pet dogs in Guangzhou, southern China, was investigated between January and December, 2009. A total of 3,977 pet dogs admitted to animal hospitals were examined for the presence of S. scabiei using a parasitological approach. The average prevalence of S. scabiei infection in pet dogs is 1.18% (95% confidence interval (CI): 0.85–1.52%). The prevalence of S. scabiei was higher in winter (1.42%; 95% CI: 0.29–2.55%), summer (1.39%; 95% CI: 0.83–1.96%), and autumn (1.1%; 95% CI: 0.53–1.68%) than in spring (0.63%; 95% CI: 0.02–1.25%). Furthermore, the prevalence of S. scabiei was the highest in Pekingese (21.88%; 95% CI: 7.55–36.2%), followed by Papillon (5.26%; 95% CI: 0–11.06%) and Bichon Frise (3.19%; 95% CI: 0–6.75%). The results of the present investigation indicate that S. scabiei infection is prevalent in pet dogs in Guangzhou, China, which provides relevant “baseline” data for conducting control strategies and measures against scabies in this region and elsewhere in China. To our knowledge, this is the first comprehensive report of S. scabiei prevalence in pet dogs in China

Identification and characterization of microRNAs in Clonorchis sinensis of human health significance

<p>Abstract</p> <p>Background</p> <p><it>Clonorchis sinensis </it>is a zoonotic parasite causing clonorchiasis-associated human disease such as biliary calculi, cholecystitis, liver cirrhosis, and it is currently classified as carcinogenic to humans for cholangiocarcinoma. MicroRNAs (miRNAs) are non-coding, regulating small RNA molecules which are essential for the complex life cycles of parasites and are involved in parasitic infections. To identify and characterize miRNAs expressed in adult <it>C. sinensis </it>residing chronically in the biliary tract, we developed an integrative approach combining deep sequencing and bioinformatic predictions with stem-loop real-time PCR analysis.</p> <p>Results</p> <p>Here we report the use of this approach to identify and clone 6 new and 62,512 conserved <it>C. sinensis </it>miRNAs which belonged to 284 families. There was strong bias on families, family members and sequence nucleotides in <it>C. sinensis</it>. Uracil was the dominant nucleotide, particularly at positions 1, 14 and 22, which were located approximately at the beginning, middle and end of conserved miRNAs. There was no significant "seed region" at the first and ninth positions which were commonly found in human, animals and plants. Categorization of conserved miRNAs indicated that miRNAs of <it>C. sinensis </it>were still innovated and concentrated along three branches of the phylogenetic tree leading to bilaterians, insects and coelomates. There were two miRNA strategies in <it>C. sinensis </it>for its parasitic life: keeping a large category of miRNA families of different animals and keeping stringent conserved seed regions with high active innovation in other places of miRNAs mainly in the middle and the end, which were perfect for the parasite to perform its complex life style and for host changes.</p> <p>Conclusions</p> <p>The present study represented the first large scale characterization of <it>C. sinensis </it>miRNAs, which have implications for understanding the complex biology of this zoonotic parasite, as well as miRNA studies of other related species such as <it>Opisthorchis viverrini </it>and <it>Opisthorchis felineus </it>of human and animal health significance.</p

Prevalence of Clonorchis sinensis infection in dogs and cats in subtropical southern China

<p>Abstract</p> <p>Background</p> <p>Clonorchiasis, caused by <it>Clonorchis sinensis</it>, is one of the major parasitic zoonoses in China, particularly in China's southern Guangdong province where the prevalence of <it>C. sinensis </it>infection in humans is high. However, little is known of the prevalence of <it>C. sinensis </it>infection in its reservoir hosts dogs and cats. Hence, the prevalence of <it>C. sinensis </it>infection in dogs and cats was investigated in Guangdong province, China between October 2006 and March 2008.</p> <p>Results</p> <p>A total of 503 dogs and 194 cats from 13 administrative regions in Guangdong province were examined by post-mortem examination. The worms were examined, counted, and identified to species according to existing keys and descriptions. The average prevalences of <it>C. sinensis </it>infection in dogs and cats were 20.5% and 41.8%, respectively. The infection intensities in dogs were usually light, but in cats the infection intensities were more serious. The prevalences were higher in some of the cities located in the Pearl River Delta region which is the most important endemic area in Guangdong province, but the prevalences were relatively lower in seaside cities.</p> <p>Conclusions</p> <p>The present investigation revealed a high prevalence of <it>C. sinensis </it>infection in its reservoir hosts dogs and cats in China's subtropical Guangdong province, which provides relevant "base-line" data for conducting control strategies and measures against clonorchiasis in this region.</p

WITHAFERIN A INDUCES APOPTOSIS IN RAT C6 GLIOMA CELLS THROUGH REGULATING NF- ΚB NUCLEAR TRANSLOCATION AND ACTIVATION OF CASPASE CASCADE

Background: The demand for the chemopreventive drug from the plant source is increasing in recent times, owing to its various biological activities without any adverse effect. The intention of this current study was to examine the antiglioma effect of Withaferin A (WFA) on C6 glioma cell line model. Materials and Methods: C6 glioma cells were administrated with different concentration of WFA (50, 100, 200 and 500 μg/mL) and DMSO (control) group to examine its anti-proliferative, anti-inflammatory and pro-apoptotic activities. Results: Treatment with WFA showed a significant decline in the glioma cell count in a dose-dependent manner and thus proving its anti-proliferative effect. Similarly, inflammatory markers were also substantially lowered upon treatment with different concentration of WFA. However, DNA fragmentation and apoptotic markers like Caspase-3 and 9 were concomitantly enhanced after co-cultured with different concentration of WFA and thus exhibiting its cytotoxicity efficacy. Furthermore, the protein expression of Bcl2 and Bax were markedly downregulated and upregulated respectively; upon treatment with WFA on C6 glioma cells. Conclusion: The outcome of this study evidently demonstrates that C6 glioma cells co-cultured with increased concentration of WFA, showed an anti-proliferative, anti-inflammatory and pro-apoptotic effect in a dose-dependent fashion

Both low and high levels of low-density lipoprotein cholesterol are risk factors for diabetes diagnosis in Chinese adults

Aims: This study aimed to investigate whether both high and low levels of low-density lipoprotein cholesterol (LDL-C), i.e., hypercholesterolemia and hypocholesterolemia, were associated with diabetes in Chinese adults. Methods: This cross-sectional study included 22,557 Chinese adults. The LDL-C reference interval was determined from a healthy sub-cohort. Associations between hypocholesterolemia or hypercholesterolemia with diabetes were analyzed using binary logistic regression. Results: The LDL-C reference interval was 1.48–3.77 mmol/L (57.23–145.78 mg/dL). Therefore, hypocholesterolemia, normocholesterolemia, and hypercholesterolemia were defined as an LDL-C concentration of 3.77 mmol/L, respectively. Prevalence of diabetes was higher in people with hypocholesterolemia or hypercholesterolemia than that in people with normocholesterolemia. Hypocholesterolemia was associated with an increased multivariable-adjusted risk for diabetes diagnosis (odds ratio, 1.57; 95% confidence interval, 1.18–2.08), and so was hypercholesterolemia (odds ratio, 1.29; 95% confidence interval, 1.10–1.51). The results remained significant after exclusion of those who took lipid-lowering drugs from the analysis. Conclusions: This study demonstrated that both low and high levels of LDL-C were associated with a higher risk of diabetes diagnosis. Patients with either high or low LDL-C may need to be closely monitored for the risk of diabetes