Multiplex serology of Helicobacter pylori antigens in detection of current infection and atrophic gastritis - A simple and cost-efficient method.

INTRODUCTION: Helicobacter pylori express a large array of antigens, each of which is duly responsible for successful colonization and pathogenesis. Here, we have studied host serum antibody responses to four of its immunodominant antigens in association with the infection status and the resulting clinical outcomes. METHODS: For this purpose, four individual H. pylori proteins (UreB, CagA, Tip-α and HP0175) were produced in recombinant forms. Serum antibody responses of 246 (75 GC and 171 NUD) patients, against the above antigens, were evaluated by multiplex immunoblotting. The associations between the resulting data and the infection status, as well as clinical outcomes were evaluated using logistic regression models. RESULTS: Serum antibodies to all four recombinant antigens increased the chances of detecting screening ELISA-positive subjects, in an escalating dose-dependent manner, ranging from 2.6 (1.5-4.7) for HP0175 to 14.3 for UreB (4.3-50.7), exhibiting the lowest and highest odds ratios, respectively (PAdj ≤ 0.001), such that 98.2% of the subjects with antibodies to all four antigens, were also positive by the screening ELISA (P < 0.0001). Among the screening ELISA-positive subjects, the three antigens of CagA, Tip-α, and HP0175 were able to segregate current from past H. pylori infection (P < 0.05). Accordingly, subjects with antibodies to one or more antigen(s) were at 5.4 (95% CI: 1.8-16.4) folds increased chances of having current infection, as compared to triple negatives (PAdj = 0.003). In reference to the clinical outcomes, those with serum antibodies to CagA were more prevalent among gastric cancer, as compared to NUD patients (ORAdj: 5.4, 95% CI: 2.4-12.2, PAdj < 0.0001). When NUD patients were categorized according to their histopathologic status, multiple antigen analysis revealed that subjects with serum antibodies to one or more of the 3 current infection-positive antigens (CagA, Tip-α, and HP0175) were at 9.7 (95% CI: 2.1-44.9, P = 0.004) folds increased risk of atrophic gastritis, in reference to triple negatives. CONCLUSION: The non-invasive multiplex serology assay, presented here, was able to not only detect subjects with current H. pylori infection, it could also screen dyspeptic patients for the presence of gastric atrophy. This simple and cost-efficient method can supplement routine screening ELISAs, to increase the chances of detecting current infections as well as atrophic gastritis.

Comparison the salivary streptococcus mutans levels between caries-active and caries-free children from Birjand, Iran: A case-control study.

Objective: Dental plaque bacteria, including Streptococcus mutans (SM), play a role in the pathogenesis of the dental caries. There are conflicting results regarding the association of salivary SM level and dental caries susceptibility. Our aim was to compare salivary SM levels in colony-forming units (CFU) between children with active caries and caries-free children in Birjand, Iran. Methods: This case-control study included 61 six-year-old children referred to health centers in Birjand city, Iran, in 2022. The children were divided into two groups: case (dmft/DMFT>0 with active caries) (including 31 children) and control (dmft/DMFT = 0 [caries-free]) (including 30 children). Demographic information and dental history were recorded. Oral examinations were also performed by the dentist. Unstimulated saliva samples were collected from children. The number of salivary SM colonies was determined using the microbial culture and confirmed using the polymerase chain reaction (PCR). The data were analyzed using Chi-square and T-tests at a significance level of p < 0.05. Results: The mean number of SM colonies was 126.24 ± 92.78 CFU/ml and 92.38 ± 75.34 CFU/ml in case and control groups, respectively. No significant difference was found in salivary SM levels between case and control groups (P = 0.125). No significant association was observed between caries experience with gender (P = 0.363), type of school (public/private) (P = 0.296), receiving oral health education (P = 0.072) and frequency of tooth brushing (P = 0.935). The mean gingival index (P = 0.001) and plaque index (P = 0.025) in case group were significantly higher than control group. Conclusion: There is no significant difference in salivary SM levels between caries-active and caries-free children in Birjandi children.

The association between high-risk human papillomavirus and oral lichen planus.

OBJECTIVES: Oral lichen planus (OLP) is a cell-mediated inflammatory mucosal disorder and is classified as an oral potentially malignant disorder. Some research has shown that apoptosis in OLP cells is similar to a viral infection such as human papillomavirus (HPV). So, the aim of this case-control study was to investigate the association of high-risk HPV with OLP. MATERIAL AND METHODS: DNA was extracted from 25 formalin-fixed, paraffin-embedded (FFPE) OLP tissues and 25 FFPE normal oral tissues as case and control groups, respectively. The presence of high-risk HPV16 and HPV18 DNA was investigated by polymerase chain reaction (PCR). p-value<.05 was considered significant. RESULTS: Twelve samples (48%) of OLPs were positive for HPV16, compared with six samples (24%) of controls; although the difference was not significant, it was borderline (p = .07). Three samples (12%) of OLPs were positive for HPV18 compared with one sample (4%) of controls; the difference was not significant (p = .3). The total frequency of both high-risk HPV were 14 samples (56%) of OLPs and 7 samples (28%) of controls; there was a significant association between the high-risk HPV and OLP (p = .04). High-risk HPVs was more prevalent in erosive-atrophic (EA) form of OLP as compared to non-EA form, although the difference was not significant (p = .13). CONCLUSIONS: The results suggest a significant association between high-risk HPVs and OLP.

Morphological and Molecular Descriptions of Macracanthorhynchus ingens (Acanthocephala: Oligacanthorhynchidae) Collected from Hedgehogs in Iran.

Aim: Limited data exist on acanthocephalan infections of hedgehogs in the world. Our objective was to investigate the prevalence and distribution of Macracanthorhynchus ingens infection in hedgehogs between August 2021 and March 2022 (n = 30) in the east of Iran. Methods: At first, infection with M. ingens was diagnosed based on morphologic features of the adults such as body length, proboscis, and hooks. Spindle-shaped eggs (mean length, 99.1 microns; mean width, 60.1 microns) were obtained from the body cavity of gravid female specimens. Results: The molecular analysis based on 18S rDNA and COX 1 genes confirmed the morphological identification of isolated M. ingens. The prevalence of M. ingens in our sample was 13.3% with 1-10 worms per infected host. Conclusion: In this study, we identify M. ingens as zoonotic species in hedgehog carcasses for the first time that passed eggs and adult worms, indicating parasite maturation and reproduction. There are a few studies on acanthocephalans in Iran. Therefore, more comparative studies are needed to determine the status of these species.

The Efficacy of Hydroalcoholic Extracts of Prosopis farcta Against Leishmania major

Objective: Cutaneous leishmaniasis caused by Leishmania major (L. major) is an endemic disease in Iran. The current reference drugs, including Glucantime, possess high toxicity in addition to some side-effects. Therefore, there is a growing interest in exploring biomedical plants. The goal of the present study was to evaluate the anti-leishmanial activity and cytotoxicity of hydroalcoholic extracts from Prosopis farcta (P. farcta) over promastigote and amastigote forms. Methods: This study was performed at the Iran Birjand University of Medical Sciences, during the year 2019. In this study, the hydroalcoholic extracts of the stems, leaves (LE) and fruits (FE) of P. farcta were obtained. The anti-leishmanial activity was assessed against leptomonad promastigotes and intracellular amastigotes of L. major. The cytotoxicity of these extracts was determined in murine macrophages. Results: The FE and LE of P. farcta demonstrated a significant leishmanicidal effect against L. major promastigotes with an IC50 of 0.9 mg/mL and 1.1 mg/mL, respectively. The FE showed the most anti-leishmanial activity and presented with the highest index of selectivity (SI=14.6) as an anti-leishmanial product. Infected macrophages treated using the FE showed a reduction in parasite burden by 97.3%. Conclusion: The results of the present study demonstrated the leishmanicidal activity of P. farcta on both promastigotes and intracellular amastigotes. There is a need for performing comprehensive studies on relevant animal models and to access the effects of active components of P. farcta extract on the growth of L. major.

The Inhibitory Effect of Mouse Gastric DNA on Amplification of Helicobacter pylori Genomic DNA in Quantitative PCR

Background: Quantitation of Helicobacter pylori (Hp) in the gastric tissue is essential for assessment of vaccination/therapeutic regimens. Materials & Results: Here, the inhibitory effect of mouse gastric DNA (MgDNA) on amplification of Hp genomic DNA (HpDNA) was evaluated by spiking HpDNA with serial dilutions of MgDNA, which yielded concentrations of >10 ng/µl and >0.63-10 ng/µl of MgDNA, as inhibition and interference zones, respectively. Mice were then inoculated with varying doses of Hp and assessed at the inhibition-free concentration of 0.63 ng/µl. The average Hp copy numbers per microgram of gastric tissue discriminated mice having received high vs. low dose inoculums (p < 0.001). Secondly, Hp copy numbers were quantitated in immunized mice, which demonstrated significantly lower numbers, in reference to controls (p = 0.006). Conclusion: Our method, bypassing the inhibition and/or interference imposed by MgDNA, was able to quantitate gastric tissue-colonizing Hp, segregating mice inoculated with low vs. high doses of Hp, as well as those immunized from controls.