Methylation and Polymorphism in CDH1 Gene Promoter Among Patients with Diffuse Gastric Cancer.

Background: The promoter methylation and single nucleotide polymorphisms (SNPs) affect the transcription activity of cancer-related genes in several cancers including diffuse gastric cancer (DGC). Here we aimed to evaluate the promoter methylation status and the rs16260 at the promoter region of the CDH1 gene in DGC. Methods: This case-control study was performed of 48 formalin-fixed paraffin-embedded (FFPE) blocks of DGC patients and 41 fresh frozen tissue samples of healthy individuals. Methylation status was evaluated using methylation-specific polymerase chain reaction (PCR) and the rs16260 at the promoter region of the CDH1 gene was assessed using PCR and sequencing method. Results: The occurrence of methylation at the promoter region of the CDH1 gene in DGC patients was significantly higher than control samples (P < 0.0001). The methylated status was significantly associated with the poor differentiated histological type of DGC (P = 0.0428). The frequency of AC genotype and the A allele in DGC patients was significantly higher than the control subjects (P = 0.006 and 0.003, respectively). Conclusions: Here we showed that methylation at the CDH1 promoter may contribute to the DGC development, and also the AC genotype was associated with the risk of DGC.

Geo-climatic factors and prevalence of chronic toxoplasmosis in pregnant women: A meta-analysis and meta-regression.

In this study, we evaluated the effects of geo-climatic parameters and other potential risk factors on the prevalence of chronic toxoplasmosis (CT) in pregnant women. We searched PubMed/MEDLINE, Web of Science, EMBASE, Scopus, and SciELO databases for seroepidemiological studies published between January 1988, and February 2021. We performed meta-analysis and meta-regression by using a random effect model to synthesize data. A total of 360 eligible datasets, including 1,289,605 pregnant women from 94 countries, were included in this study. The highest and lowest prevalence rates were estimated for latitudes of 0-10° (49.4%) and ≥50° (26.8%); and for the longitude of 80-90° (44.2%) and 110-120° (7.8%), respectively. Concerning climatic parameters, the highest and lowest prevalence rates were estimated in regions with the mean relative humidities of >80% (46.6%) and <40% (27.0); annual precipitation between 1000 and 1500 mm (39.2%) and 250-500 mm (26.8%); and mean annual temperature of 20-30 °C (36.5%), and <7 °C (24.9%), respectively. Meta-regression analyses indicated significant increasing trends in prevalence of CT in pregnant women with decrease in geographical latitude (coefficient, = -0.0035), and geographical longitudes (C = -0.0017). While it was positively associated (P < 0.01) with the mean environmental temperature (C = 0.0047), annual precipitation (C = 0.000064), and mean relative humidity (C = 0.002). Our results highlighted various effects of environmental parameters on the prevalence of CT. Therefore, different regions in the world may benefit from different types of interventions, and thus, novel preventive measures in a region should be developed according to local climate, agricultural activities and people culture.

Toxocara infection/exposure and the risk of schizophrenia: a systematic review and meta-analysis.

Toxocariasis is an important neglected disease that could have a role in the development of some neurological disorders, such as schizophrenia (SCZ). We performed the first systematic review and meta-analysis on observational studies to evaluate the possible association between Toxocara infection/exposure and SCZ. Systematic searching (PubMed, Scopus, Web of Science and Google Scholar) was performed, up to 15 August 2020, to identify the relevant studies. We used a random effects meta-analysis model to generate the pooled OR and 95% CIs. The research process resulted in the identification of six studies related to the subject of interest entailing 411 SCZ patients and 2047 controls. Our results demonstrated that the SCZ patients (15%; 95% CI 6.8 to 29.9%) had higher pooled seroprevalence of Toxocara infection/exposure than the control group (3.3%; 95% CI 1.1 to 9.4%), showing a significant association (pooled OR, 4.06; 95% CI 1.14 to 14.43). Our findings suggested that Toxocara infection/exposure might have an important but neglected contribution to SCZ development. However, further well-designed and large-scale observational studies are needed to illuminate this association. Because many other environmental and genetic factors may be strong confounders, more mechanistic/experimental studies in animal models are also necessary to further define the mechanism of this association.

Toxoplasma oocysts in the soil of public places worldwide: a systematic review and meta-analysis.

Toxoplasmosis is a zoonotic and cosmopolitan infection. Although a few studies have evaluated the prevalence of Toxoplasma oocysts in the soil of public places, the present study was conducted to provide insights into environmental contamination levels and its potential transmission to humans on a global scale. A systematic search was conducted using bibliographic databases through 30 August 2020. A random effects model was utilized to estimate pooled prevalence with 95% confidence intervals (CIs). Subgroup analysis and meta-regressions were also performed on the geographical and environmental parameters. Finally, 22 articles, wherein 15 420 soil samples were examined, met the systematic review and meta-analysis requirements. The mean pooled prevalence of Toxoplasma oocysts was estimated at 16% (95% CI 10 to 26) in public places. The estimated prevalences in Europe, South America, Asia and North America were 23% (95% CI 4 to 65), 22% (95% CI 18 to 26), 15% (95% CI 0.06 to 33) and 8% (95% CI 0.00 to 97), respectively. An increasing trend was observed in the prevalence of Toxoplasma oocysts with increasing latitude (41-56°), decreasing longitude (0-40°) and increasing relative humidity (≥76%). Loop-mediated isothermal amplification and polymerase chain reaction methods revealed the highest and lowest prevalence rates, respectively, in the detection of Toxoplasma oocysts. Awareness of the health authorities and people about Toxoplasma prevalence in the soil of public places and its risk factors is of great importance to developing effective strategies to prevent infection.

The prevalence of latent and acute toxoplasmosis in HIV-infected pregnant women: A systematic review and meta-analysis.

OBJECTIVE: HIV in pregnancy is not only important for mother-to-child HIV transmission, but also it assumes additional importance because HIV increases susceptibility to opportunistic infections, leading to increased morbidity and mortality in mothers and neonates. Toxoplasmosis is one of the most important opportunistic infections in HIV-infected pregnant women. The present study was undertaken to assess the prevalence of latent toxoplasmosis (LT) and acute toxoplasmosis (AT) infection in HIV-infected pregnant women. METHODS: PubMed/MEDLINE, Scopus, Web of Science, EMBASE and SciELO were searched to identify relevant studies. A random-effects model was used to estimate the overall and subgroup-pooled prevalences across studies. Heterogeneity between studies was assessed via the I2 test. RESULTS: A total of 14 articles that included 3256 subjects in nine countries met the inclusion criteria. The overall prevalence rates of LT and AT in HIV-infected pregnant women were 45.7% (95% CI, 32.3-59.7%) and 1.1% (95% CI, 0.4-3.2%), respectively. The findings indicate that, worldwide, approximately 559,000 and 13,450 HIV-infected pregnant women are affected by LT and AT, respectively. From this review, it is estimated that approximately 3432 babies annually could be born with congenital toxoplasmosis (CT) from HIV-infected pregnant mothers. CONCLUSIONS: The present study indicates that a large number of HIV-infected mothers are affected by LT and AT. This can lead to adverse complications such toxoplasmic encephalitis in mothers and CT in neonates. Our results suggest a need for screening programs using well-validated diagnostic platforms for both LT and AT for all HIV-infected pregnant women.

Designing a Recombinant Multi-Epitope Antigen of Echinococcus granulosus to Diagnose Human Cystic Echinococcosis.

BACKGROUND: Cystic echinococcosis can cause severe disease and probable death in humans. Epitopes of its antigens play a key role in the sensitivity and specificity of immunodiagnostic tests. METHODS: Epitope prediction software programs predict the most antigenic linear B-cell epitopes of AgB (8 kD), Ag5, and Ag95. Six such epitopes were predicted and connected by "Gly-Ser" linker and synthesized. The purity of the concentrated recombinant multi-epitope protein was assessed by 15% SDS-PAGE. Overall, 186 serum samples were collected from the Loghman Hakim Hospital and different laboratories, Tehran, Iran, from July 2016 to February 2017. Patients infected with hepatic hydatid cysts, patients infected by other parasites and viruses, and healthy individuals were used to detect the anti-CE IgG using recombinant multi-epitope protein. RESULTS: Forty-one samples out of 43 cases of hydatidosis were diagnosed correctly as positive, and two were negative. In addition, six negative cases of healthy individual group were diagnosed as positive and negative with rMEP-ELISA and the commercial kit, respectively. Therefore, these six samples were considered as false positive using our method. In addition, a diagnostic sensitivity of 95.3% (95% CI, 84.19% to 99.43%) and a specificity of 95.0% (95% CI, 89.43% to 98.14%) were obtained using optimum cutoff value (0.20). The sensitivity and specificity of the commercial kit was 100%. CONCLUSION: Our findings showed high diagnostic accuracy of the ELISA test using the developed recombinant protein, which encourages the use of this recombinant multi-epitope protein for rapid serological diagnosis of hydatidosis.

Global Prevalence Estimates of Toxascaris leonina Infection in Dogs and Cats.

Toxascaris leonina is an ascaridoid nematode of dogs and cats; this parasite affects the health of these animals. This study estimated the global prevalence of Ta. leonina infection in dogs and cats using random effects meta-analysis as well as subgroup, meta-regression and heterogeneity analyses. The data were stratified according to geographical region, the type of dogs and cats and environmental variables. A quantitative analysis of 135 published studies, involving 119,317 dogs and 25,364 cats, estimated prevalence rates of Ta. leonina in dogs and cats at 2.9% and 3.4%, respectively. Prevalence was highest in the Eastern Mediterranean region (7.2% for dogs and 10.0% for cats) and was significantly higher in stray dogs (7.0% vs. 1.5%) and stray cats (7.5% vs. 1.8%) than in pets. The findings indicate that, worldwide, ~26 million dogs and ~23 million cats are infected with Ta. leonina; these animals would shed substantial numbers of Ta. leonina eggs into the environment each year and might represent reservoirs of infection to other accidental or paratenic hosts. It is important that populations of dogs and cats as well as other canids and felids be monitored and dewormed for Ta. leonina and (other) zoonotic helminths.

Intra-Species Diversity of Leishmania major and L. tropica from Clinical Isolates of Cutaneous Leishmaniasis in Southwest Iran Inferred by ITS1-rDNA.

BACKGROUND: Cutaneous leishmaniasis (CL) as a public health concern is increasingly circulating by causative agents of Leishmania tropica and L. major in Iran. As regard to recent treatment failure and controlling problems, the accurate elucidation of heterogeneity traits and taxonomic status of Leishmania spp. should be broadly addressed by policymakers. This study was designed to determine the genetic variability and molecular characterization of L. major and L. tropica from Iranian CL patients. METHODS: One hundred positive Giemsa-stained slides were taken from clinical isolates of CL at Pol-e-Dokhtar County, Southwest Iran, from May 2014 to Sep 2016. DNAs were directly extracted and amplified by targeting ribosomal internal transcribed spacer (ITS) gene following microscopic observation. To identify Leishmania spp. amplicons were digested by restriction enzyme HaeIII subsequent PCR-RFLP technique. To reconfirm, the isolates were directly sequenced to conduct diversity indices and phylogenetic analysis. RESULTS: Based upon the RFLP patterns, 84 and 16 isolates were explicitly identified to L. tropica and L. major respectively. No co-infection was found in clinical isolates. The high genetic diversity of L. tropica (Haplotype diversity 0.9) was characterized compared to L. major isolates (Hd 0.476). The intra-species diversity for L. tropica and L. major isolates corresponded to 3%-3.9% and 0%-0.4%, respectively. CONCLUSION: Findings indicate the L. tropica isolates with remarkable heterogeneity than L. major are predominantly circulating at Pol-e-Dokhtar County. Occurrence of high genetic variability of L. tropica may be noticed in probable treatment failure and/or emerging of new haplotypes; however, more studies are warranted from various geographic regions of Southwest Iran, using large sample size.

A Survey on the Adjuvant Role of Naloxone Alone or Combined with Alum in Vaccination Against Fasciolosis in BALB/c Mice.

BACKGROUND: Fasciolosis is a zoonotic parasitic disease imposing a heavy load of livestock losses worldwide. PURPOSE: We aimed to evaluate immune-stimulatory effects of naloxone (NLX), an opioid receptor antagonist, in combination with alum in mice vaccinated with excretory-secretory antigens (E/S) of Fasciola hepatica. METHODS: 8-week-old female BALB/c mice were subcutaneously vaccinated using E/S antigens of F. hepatica. Experimental groups (14 mice per group) included: vaccine (E/S antigen), alum vaccine (E/S antigen plus alum), NLX vaccine (E/S antigen plus NLX), and alum-NLX vaccine (E/S antigen plus a mixture of alum-NLX). The control group was infused with PBS. Lymphocyte proliferation and the levels of IFN-γ, IL-4, IgG2a, IgG1, and total IgG were measured. RESULTS: Mice vaccinated with NLX or alum-NLX adjuvants showed significantly higher rates of lymphocyte proliferation, IFN-γ, total IgG, and IgG2a levels. The mice that were injected with alum showed a significantly higher concentration of IL-4. Ratios of IFN-γ/Il-4 and IgG2a/IgG1 were significantly higher in the NLX and alum-NLX groups in comparison with the groups vaccinated either with alum or without any adjuvant. A significantly higher protection rate (62.5%) was seen in mice vaccinated with the alum-NLX adjuvant compared to the other groups. CONCLUSION: NLX can be effective in conferring cellular immunity and protection against F. hepatica. It is recommended to consider this agent as a potential adjuvant in vaccines against fasciolosis.

Super Infection of Cutaneous Leishmaniasis Caused by Leishmania major and L. tropica to Crithidia fasciculata in Shiraz, Iran.

BACKGROUND: The aims of the current study were to determined present status of CL in Shiraz City, identify the causative species of Leishmania and conduct phylogenetic evaluations in detected parasites. METHODS: This study was conducted on 70 individuals with suspected CL that referred to the major health centers of Shiraz (Valfajr), Fars province, Iran, from Sep 2016 to Jul 2017. DNA was extracted from cultured Leishmania promastigotes and PCR-RFLP were performed using ITS1-rDNA gene. RESULTS: Overall, 39 male (55.70%) and 31 (44.30%) female were found to be positive microscopically. All of direct examined positive samples were confirmed to be positive for Leishmania spp. DNA. Based upon the PCRRFLP patterns and phylogenetic analysis, 46 (65.72%), 17 (24.28%) and 7 (10%) isolates were clearly identified as L. major, L. tropica and C. fasciculata, respectively. CONCLUSION: The dominat detected species in Shiraz City was L. major and L. tropica, respectively. CL has high prevalence in Shiraz City; therefore, more studies on leishmaniasis in the natural vectors and also reservoirs infection in this region is exceedingly recommended. Skin leisons due to C. fasciculata, was described for the first time in Iran (Shiraz City).

Facile green synthesis of silver nanoparticles using Berberis vulgaris leaf and root aqueous extract and its antibacterial activity.

Most recently, silver nanoparticles due to antibacterial properties have been considered in medical science. So the aim of the study was green synthesis of silver nanoparticles using Berberis vulgaris leaf and root aqueous extract and its antibacterial activity. After collection, identification and extraction of Berberis vulgaris was performed production of silver nanoparticles. In the study effect of parameters such as AgNO3 concentration (0.5, 1, 3, 10 mM), aqueous extract (3, 5, 10, 15, 30 mL) and contact time (1, 2, 6, 12, 24 h) were investigated in the synthesis of nanoparticles and also the antibacterial effect of these nanoparticles was studied on Escherichia coli and Staphylococcus aureus bacteria by Disk diffusion test and Minimum Inhibitory Concentration test (MIC). According to XRD results and analysis of TEM, nanoparticles have spherical shapes and size of 30 to 70 nm. On the other hand antibacterial tests showed these nanoparticles have more antibacterial activity more than other extracts. Result showed the biosynthesis of silver nanoparticles using aqueous extract of Berberis vulgaris is a clean, inexpensive and safe method that has not been used any toxic substance and consequently does not side effects and this nanoparticles has a high antibacterial activity.

Naltrexone; as an efficient adjuvant in induction of Th1 immunity and protection against Fasciola hepatica infection.

Toxic effects of available therapeutics are major drawbacks for conventional management approaches in parasitic infections. Vaccines have provided a promising opportunity to obviate such unwanted complications. In present study, we examined immune augmenting capacities of an emerging adjuvant, Naltrexone, against Fasciola hepatica infection in BALB/c mice. Seventy BALB/c mice were divided into five experimental groups (14 mice per group) including 1- control (received PBS), 2- vaccine (immunized with F. hepatica E/S antigens), 3- Alum-vaccine (immunized with Alum adjuvant and E/S antigens), 4- NLT-vaccine (immunized with NLT adjuvant and E/S antigens), and 5- Alum-NLT-vaccine (immunized with mixed Alum-NLT adjuvant and E/S antigens). Lymphocyte stimulation index was assessed by MTT assay. Production of IFN-γ, IL-4, IgG2a and IgG1 was assessed by ELISA method. Results showed that NLT, either alone or in combination with alum, can induce immune response toward production of IFN-γ and IgG2a as representatives of Th1 immune response. Also, using this adjuvant in immunization experiment was associated with significantly high proliferative response of splenocytes/lymphocytes. Utilization of mixed Alum-NLT adjuvant revealed the highest protection rate (73.8%) in challenge test of mice infected with F. hepatica. These findings suggest the potential role of NLT as an effective adjuvant in induction of protective cellular and Th1 immune responses against fasciolosis.

Frequency of Intestinal Parasites among Zoo Animal by Morphometric Criteria and First Report of the Bivitellobilharzia nairi from Elephant (Elephasmaximus maximus) in Iran.

BACKGROUND: Intestinal parasitic infections are major causative agents of wildlife health complications among different parts of the world. This study aimed to investigate the gastro-intestinal parasites in feces of the zoo animals based on parasitological and morphometric criteria. METHODS: One hundred fresh fecal samples were collected from 35 species of animal lived in Eram park zoo, Tehran, Central Iran during Oct 2015 to Jun 2015. All collected samples were examined by microscopic observation following direct wet mount preparation (normal saline and Lugol's iodine), formalin-ether concentration, and permanent staining. The morphometric aspects of the recovered eggs were surveyed with the aid of Camera Lucida (×400). RESULTS: 65.7% (23/35) of zoo animal species were infected with intestinal parasites. The superfamily Trichostrongyloidea (6/16) and Strongylus sp. (16/4) were the most prevalent helminthic infections, while Blastocystis sp. (6/14), Entamoeba cyst (3/14) and Eimeria sp. (3/14) were the common protozoan parasites. For the first time, Bivitellobilharzia nairi egg was identified an elephant at Iran. Intestinal parasitic infections were apparently circulating among animals of the Eram park zoo. CONCLUSION: Identified parasitic infections can consider as a threatening source to visitors and workers' health that have contact with animals or their feces. Therefore, the effectual preventive strategies should be addressed to determine the risk factors, mechanisms of cross-transmission of parasite, the importance of applying the hygienic practices and well adjusting deworming programs for the animals, zoo workers and visitors.