Chlamydia pneumonia infection and risk of multiple sclerosis: A meta-analysis.

BACKGROUND: The role of infectious agents, including Chlamydia pneumoniae (Cpn), in the development of multiple sclerosis (MS), is still a matter of major contention. OBJECTIVE: This meta-analysis study aimed to assess the actual involvement of Cpn in MS development. METHODS: We undertook a search of international scientific databases to identify eligible studies. We used a random-effects meta-analysis model (REM) to generate the pooled odds ratio (OR) and 95% confidence intervals (CIs). Heterogeneity was calculated using the I2 statistic. Sensitivity and subgroup analyses were applied to assess the effects of study characteristics and socio-demographic variables on the pooled OR. RESULTS: We identified 37 studies comprising 51 datasets that satisfied the inclusion criteria. Considering diagnostic methods for Cpn, 26 and 25 datasets used PCR- and serological-based methods, respectively. In PCR-based datasets, REM showed a significant positive association between Cpn infection and the development of MS (OR, 5.29; 95% CI, 3.12-8.97), while a non-significant positive association was achieved in serological-based datasets (OR, 1.34; 95% CI, 0.88-2.03). In subgroup analyses on PCR-based datasets, results were significant for both CSF (OR, 5.70) and serum (OR, 4.84) samples; both healthy (OR, 16.11) and hospital-based (OR, 2.88) controls; and both moderate (OR, 5.14) and high (OR, 5.48) quality studies. In serological-based datasets, only those that used CSF samples yielded significant results (OR, 3.41). CONCLUSIONS: Our findings verify the significant positive relationship between Cpn infection and MS. We advocate prospective cohort studies with lifelong follow-ups and also experimental studies to better understand the role of Cpn in MS development.

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.

Cryopreservation of Echinococcus granulosus Protoscoleces.

BACKGROUND: The main objective of the current study was to investigate on the cryopreservation of protoscoleces of Echinococcus granulosus, a causative agent of cystic hydatidosis in man. METHODS: This study was conducted on isolated protoscoleces from hydatid cysts infected livers collected from slaughterhouse of Tehran, Iran in 2016. Viability of protoscoleces was evaluated by dye test. Cryopreservation of isolated protoscoleces in the presence of Dimethylsulfoxide (DMSO) and glycerol using a three-step cooling protocol involving an initial period at -20 °C, -80 °C and liquid nitrogen was performed. RESULTS: The mean viability rate of 10% DMSO and 15% glycerol were 9% and 8% respectively. The protoscoleces of Echinococcus granulosus have been successfully thawed and recovered after 6 months storage in liquid nitrogen. CONCLUSION: Cryopreservation method needs to be improved for each species of helminthes and can be useful for other immunological and laboratorial studies.

Antileishmanial activity of Urtica dioica extract against zoonotic cutaneous leishmaniasis.

BACKGROUND: Neglected parasitic diseases (NTDs) like cutaneous leishmaniasis (CL) have caused high mortality and morbidity rate in developing countries. This disease is considered as one of the six major tropical diseases, and has a great importance in HIV infected individuals as an opportunistic infection in those areas that both infections are endemic. This study evaluated the therapeutic effects of the Urtica dioica L (U. dioica) aqueous extract as an anti-leishmanial herbal drug in-vitro and in-vivo, and in addition to that, evaluated two vital immune system cytokines including gamma interferon (IFN-γ) and interleukin-4 (IL-4) plus nitric oxide (NO) and arginase activity against Leishmania major (L. major) infected mice. METHODOLOGY/PRINCIPAL FINDINGS: In-vitro anti-leishmanial activity of U. dioica aqueous extract was determined using MTT method and also Parasite Rescue Transformation Assay. Also, the footpad lesion size and parasite load in BALB/c mice infected with L. major were quantified for in-vivo assessment. Furthermore, for evaluating the immune responses, the levels of IFN-γ, IL-4, NO and arginase were measured in the BALB/c mice. These results indicated that U. dioica extract significantly reduced the L. major promastigotes viability. According to the in-vitro cytotoxicity assay of the extract on Leishmania parasites (CC50) and infected macrophages (EC50), the extract had no toxicity to the macrophages, however it efficiently killed the L. major amastigotes. In addition, the lesion size, parasite load, IL-4, and ARG were decreased in the treated infected mice, however IFN-γ and NO were significantly increased. CONCLUSIONS/SIGNIFICANCE: This study established satisfactory results in Leishmania parasite clearing both in-vivo and in-vitro. Therefore, U. dioica extract can be considered as an effective and harmless herbal compound for killing the parasite without toxicity to the host macrophages. Furthermore, it also can treat the CL by switching the mouse immune response towards a cell-mediated response (Th1); hence, it may be identified as a perfect therapeutic herbal drug for CL treatment.

Anacardic Acid Reduces Acetylation of H4K12 in Mouse Oocytes during Vitrification.

OBJECTIVE: Over the last years, vitrification has been widely used for oocyte cryopreservation, in animals and humans; however, it frequently causes minor and major epigenetic modifications. The effect of oocyte vitrification on levels of acetylation of histone H4 at lysine 12 (AcH4K12), and histone acetyltransferase (Hat) expression, was previously assessed; however, little is known about the inhibition of Hat expression during oocyte vitrification. This study evaluated the effect of anacardic acid (AA) as a Hat inhibitor on vitrified mouse oocytes. MATERIALS AND METHODS: In this experimental study, 248 mouse oocytes at metaphase II (MII) stage were divided in three experimental groups namely, fresh control oocytes (which were not affected by vitrification), frozen/thawed oocytes (vitrified) and frozen/thawed oocytes pre-treated with AA (treatment). Out of 248 oocytes, 173 oocytes were selected and from them, 84 oocytes were vitrified without AA (vitrified group) and 89 oocytes were pretreated with AA, and then vitrified (treatment group). Fresh MII mouse oocytes were used as control group. Hat expression and AcH4K12 levels were assessed by using real-time quantitative polymerase chain reaction (PCR) and immunofluoresce staining, respectively. In addition, survival rate was determined in vitrified and treatment oocytes. RESULTS: Hat expression and AcH4K12 modification significantly increased [4.17 ± 1.27 (P≤0.001) and 97.57 ± 6.30 (P<0.001), respectively] in oocytes that were vitrified, compared to the fresh oocytes. After treatment with AA, the Hat mRNA expression and subsequently H4K12 acetylation levels were significantly reduced [0.12 ± 0.03 (P≤0.001) and 89.79 ± 3.20 (P≤0.05), respectively] in comparison to the vitrified group. However, the survival rate was not significantly different between the vitrified (90.47%) and treatment (91.01%) groups (P>0.05). CONCLUSION: The present study suggests that AA reduces vitrification risks caused by epigenetic modifications, but does not affect the quality of vitrification. In fact, AA as a Hat inhibitor was effective in reducing the acetylation levels of H4K12.

Sambucus ebulus extract stimulates cellular responses in cutaneous leishmaniasis.

This is the first study aiming to determine the therapeutic effects of the Sambucus ebulus aquatic extract as an antileishmanial herbal drug and evaluate the immune responses in Leishmania major major infected BALB/c mice. The antileishmanial activity of S ebulus aquatic extract was evaluated using MTT test as well as parasite rescue and transformation assay. Footpad swelling and parasite load of infected mice were measured by several techniques. The immune responses were evaluated by measuring the levels of IFN-γ, IL-4, nitric oxide and arginase. The results indicated that S. ebulus can significantly decrease L. major promastigotes and amastigotes viability, but it was not toxic to macrophages. The lesion size, parasite burden and the level of ARG decreased in the treated infected mice, while the IFN-γ-to-IL-4 ratio and the level of NO increased significantly. Altogether, the S. ebulus extract is an effective compound for killing Leishmania parasite without excessive toxicity to the host cells and can cure the CL by switching the host immune responses towards Th1 response. Thus, it may be a perfect therapeutic option for CL treatment.

Comparison of the antibacterial effects of a short cationic peptide and 1% silver bioactive glass against extensively drug-resistant bacteria, Pseudomonas aeruginosa and Acinetobacter baumannii, isolated from burn patients.

We have already established that a short cationic peptide (CM11) has high antimicrobial activity against a number of bacterial pathogens. Considering the untreatable problem of burn infections caused by Pseudomonas aeruginosa and Acinetobacter baumannii, this study evaluated and compared antibacterial effects of the CM11 peptide and 1% silver-doped bioactive glass (AgBG) against extensively drug-resistant strains of these bacteria which were isolated from burn patients. Accordingly, the bacteria were isolated from burn patients and their antibiotic resistance patterns and mechanisms were fully determined. The isolated bacterial from patients were resistant to almost all commonly used antibiotics and silver treatment. The isolates acquired their resistance through inactivation of their porin, the overexpression of efflux pump, and beta-lactamase. CM11 peptide and 1% AgBG had minimum inhibitory concentration (MIC) of ≥ 16 µg ml-1 and ≥ 4 mg ml-1 for clinical isolates, respectively. The minimum bactericidal concentration (MBC) of peptide and 1% AgBG for resistant bacteria was ≥ 32 µg ml-1 and ≥ 4 mg ml-1, respectively. Among the clinical isolates, two P. aeruginosa isolates and one A. baumannii isolate were resistant to 1% AgBG disk. The CM11 peptide also showed high biocompatibility in vivo and no cytotoxicity against fibroblasts and adipose-derived mesenchymal stem cells in concentrations ≤ 64 µg ml-1 and ≤ 32 µg ml-1, respectively, while the safe concentration of 1% AgBG for these cells was ≤ 16 µg ml-1. In conclusion, these findings indicated that the 1% silver is not safe and effective for treatment of such infections. The data suggest that CM11 peptide therapy is a reliable and safe strategy that can be used for the treatment of burn infections caused by antimicrobial-resistant isolates. The next stage of the study will be a multicenter clinical trial.

Melatonin Modifies Histone Acetylation During In Vitro Maturation of Mouse Oocytes.

OBJECTIVES: We evaluated the effect of melatonin, as a potent antioxidant agent, on glutathione (GSH) and reactive oxygen species (ROS) levels, as well as histone H3 lysine 9 (H3K9), and H4 lysine 12 (H4K12) acetylation when added to oocytes culture medium. MATERIALS AND METHODS: In this experimental study, two in vitro and in vivo groups were used. In the in vitro group, cumulus oocyte complexes (COCs) from the ovaries of B6D2F1 mice were cultured in maturation medium containing two doses of melatonin (10-9 and 10-6 M) and without melatonin [control group treated with dimethyl sulfoxide (DMSO)] for 22-24 hour. The cumulus expansion and nuclear status were monitored by an inverted microscope. Next, COCs were isolated from the oviducts of superovulated mice and studied as the in vivo group. In in vitro and in vivo matured oocytes, GSH and ROS levels were assessed by monochlorobimane (MCB) and 2-7-dichlorodihydrofluorescein diacetate (H2DCFDA) staining, respectively. Changes in histone acetylation were examined by immunofluorescent staining with specific antibodies against acetylated H3K9 and H4K12. RESULTS: The H4K12 acetylation and ROS levels were significantly higher in the oocytes matured in the in vitro group compared to the in vivo group (P<0.05). Furthermore, glutathione levels in the in vitro group were considerably lower than that of the in vivo group (P<0.05). Melatonin at the concentration of 10-6 M had the most substantial effect on nuclear maturation and histone acetylation as well as glutathione and ROS levels in the in vitro group (P<0.05). CONCLUSIONS: Exogenous melatonin improves the competence of mouse oocytes during in vitro maturation (IVM).

Seroprevalence of IgG Antibodies against Echinococcus granulosus by ELISA Method Using Recombinant Agb in Lorestan Province, Western Iran.

BACKGROUND: Cystic echinococcosis (CE) is a zoonotic disease with global prevalence, which causes considerable health problems and economic losses throughout the world. The aim of this study was to assess the seroepidemiology of CE in Doroud City, Lorestan Province, Iran, considered a neglected endemic location. METHODS: An ELISA was performed using recombinant AgB from Apr to Jul 2015 in Lorestan Province, Western Iran. The commercial Hydatidosis IgG ELISA kit (Vircell SL, Granada, Spain) was used to confirm the obtained results. RESULTS: In the present study, out of 927 collected sera, 25 samples (2.6%) were found as seropositive for E. granulosus IgG antibodies. The prevalence of IgG antibodies against E. granulosus was significantly higher in rural areas (3.24%) than in urban area (1.20%) (P<0.001). Moreover, there was no significant relationship between age, occupation, sex, and literacy with seropositivity (P>0.05). Moreover, there was no statistically significant difference between the prevalence of CE in males (13/349, 3.72%) and females (12/553, 2.12%). With regard to occupation, farmers and ranchmen had the highest rate of infection (5.5%). There was a significant association between eating unwashed vegetables and seropositivity (P<0.001). Seropositive cases in rural areas were more than in urban areas. CONCLUSION: Since all the seropositive cases used unwashed local vegetables, the contamination may occur through the consumption of such vegetables.

Contamination of commonly consumed raw vegetables with soil transmitted helminth eggs in Mazandaran province, northern Iran.

Soil-transmitted helminth (STH) infections are responsible for significant burden of morbidity and mortality worldwide. Consumption of raw vegetables without proper washing is one of the major routes of such infections. We evaluate the prevalence of STH contamination in commonly used vegetables in Mazandaran province, northern Iran. A total of 772 fresh raw vegetables were obtained from retail markets. Each sample was divided into two groups. One group was used as the unwashed sample and the second group was washed with standard washing procedures. Then, samples were examined for helminth eggs by using standard methods. Data analysis was performed using SPSS20. The overall prevalence of STHs was 14.89% (115/772). The rate of STH contamination was significantly higher in warm seasons (20.5%, 79/386) than in cold seasons (9.32%, 36/386) among the unwashed vegetables (OR=2.50; CI 95%=1.64-3.8; P<0.001). No parasites were observed in standard washed samples (OR=271.40; CI 95%=16.84-4373.64; P<0.001). Prevalence of STH contamination was significantly higher in leafy vegetables than root vegetables (OR=1.67; CI 95%=1.09-2.55; P<0.05). The prevalence of STHs species in all the vegetables were as follows: Ascaris lumbricoides (3.36%), Trichuris trichiura (2.2%), hookworms (2.9%), Toxocara spp. (1.68%), Trichostrongylus spp. (1.55), Taenia sp. (0.9%) and Hymenolepis nana (2.2%). The results of the present study emphasized that vegetables are potential risk factor for transmission of helminth infection to human in northern Iran. It is necessary that health authorities trained the consumers to proper and standard washing of vegetables before consumption.

High-Level Expression of Immunogenic Recombinant Plasmodium vivax Merozoite Surface Protein (Pvmsp-142 kDa) in pGEX 6P1 Vector.

BACKGROUND: Detection of Plasmodium vivax specific antibodies with serological tests could be a valuable tool for epidemiological researches. Whereas P. vivax cannot be simply obtained in vitro, serological tests using total or semi-purified antigens are infrequently used. Given this restriction, the present study investigated whether recombinant P. vivax merozoite surface protein 1 (PvMSP-1 42 kDa) could be useful in detection of antibodies from the serums of a P. vivax infected person using serological tests. METHODS: Parasite DNA was extracted from blood sample of an Iranian P. vivax-infected patient. The region of PvMSP-142 kDa was amplified by PCR then cloned into pTZ57R/T vector and sequenced. The insert was sub cloned into pGEX 6P1 expression vector. Afterwards, it was transformed into E. coli BL21 and cultured massively. Sub cloning of gene was confirmed by PCR and enzyme digestion and sequencing finally. Production of recombinant protein was confirmed by SDS-PAGE. Western blot was performed by human sera to appraisal binding ability to the IgG antibodies of P. vivax infected patients. Recombinant protein was purified and estimated by Bradford assay. RESULTS: The specialty values of the Western blot determined with 10 sera from naturally infected individuals, 10 sera from healthy individuals and 7 sera from individuals with other infectious diseases. CONCLUSION: For the Iranian population, using a Western blot assay for MSP-142 recombinant protein can be used as the foundation for promotion of serological assay for the detection of P. vivax malaria such as ELISA.