Molecular epidemiology and associated risk factors of oral cavity parasites in hemodialysis patients in western Iran.

Renal disorders are able to make patients more susceptible to infections, including those caused oral cavity infections. Today, it has been proven that during end stage renal failure, hemodialysis results in acquired immune response defects through failings of humoral and cell-mediated immune. This survey was intended to assess the molecular epidemiology and associated risk factors of Entamoeba gingivalis and Trichomonas tenax as the main oral cavity protozoa in hemodialysis patients in Lorestan Province, western Iran. The investigation was performed on 73 hemodialysis patients referring to hemodialysis centers of Lorestan Province, Iran during May 2021 to February 2022. The frequency of oral cavity protozoa was investigated using microscopic and conventional polymerase chain reaction (PCR). A provided questionnaire with a number of demographical information and related risk factors was completed for each patient. The results showed that E. gingivalis and T. tenax parasites were found in 18 (24.6%) and 20 (27.4%) of the hemodialysis participants by microscopic and PCR test, respectively. Among samples, 13 (17.8%) of the hemodialysis participants were infected with E. gingivalis; whereas 7 (9.6%) of the participants were positive for T. tenax. No significant association was reported between gender, age, education, flossing, use of mouthwash, and prevalence of oral protozoa in hemodialysis participants. However, a significant correlation was observed among between living in rural regions (OR = 13.16; 95% CI = 2.64-56.81; p = 0.002), brushing teeth (OR = 8.51; 95% CI = 1.71-42.3; p = 0.009) and prevalence of oral protozoa in hemodialysis participants. The findings of these epidemiological study clearly showed the high frequency of oral cavity parasites in hemodialysis patients in Lorestan Province, Western Iran. Awareness of the main risk factors for oral cavity parasites particularly teeth brushing is necessary in refining public and oral health policies in hemodialysis patients. Consequently, dental practitioners, nephrologist, and urologist must be alert of these risk factors to carefully identify and achieve oral health concerns in hemodialysis patients to prevent the oral diseases and infections.

Anti-helminthic effects and cellular mechanisms of Astragalus ecbatanus extract against Echinococcus granulosus protoscoleces.

We aimed to investigate the in vitro and ex vivo anti-helminthic effects of Astragalus ecbatanus chloroform extract (AECE) as well as its cellular mechanisms against Echinococcus granulosus protoscoleces. In vitro and ex vivo anti-helminthic effects of AECE on E. granulosus protoscoleces were evaluated through eosin exclusion test. Effects of AECE on induction of the caspase-3 like activity, the permeability of plasma membrane were also determined. The effects of AECE against the human hepatocyte carcinoma (HepG2) and human embryonic kidney (HEK293T) cells were determined by MTT ([3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide)]) test. The findings revealed that AECE especially at 45 mg/mL displayed potent in vitro and ex vivo anti-helminthic effects against E. granulosus protoscoleces (p < 0.001). After treatment of protoscoleces with AECE, the caspase-3 enzyme activity and the rate of permeability of plasma membrane was dose dependently (P < 0.001) increased. The 50% cytotoxic concentration (CC50) value of AECE for normal and cancer cells lines 4.62 mg/mL and 1.89 mg/mL, respectively. The findings of the current study revealed the favorable in vitro and ex vivo anti-helminthic effects of AECE against protoscoleces of E. granulosus. However, further surveys in animal model are required to elucidate different aspects of this extract before use in clinical phases.

Potential effects of alpha-pinene, a monoterpene commonly found in essential oils against Toxoplasma gondii infection; an in vitro and in vivo study.

This survey designed to assess the in vitro and in vivo activity of α-pinene, a monoterpene commonly originated in essential oils on Toxoplasma gondii. The in vitro effect of various concentration of α-pinene against tachyzoites of T. gondii Rh strain was assessed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The activity of α-pinene on the stimulation of apoptosis in tachyzoites of T. gondii was also examined using the caspase 3 colorimetric activity assay. In vivo assay, mice were orally received α-pinene at 2 and 4 mg/kg/day for 14 days, then, pre-treated mice were daily tested and the rate of death was recorded. α-pinene meaningfully declined (p < 0.001) the tachyzoites viability with the IC50 value of 23.3 µg/mL. α-pinene induced the apoptosis through increasing the caspase-3 activity by 35.6%. Oral treatment with α-pinene significantly (p < 0.01) improved the survival rate infected mice with by 9th day. α-pinene + atovauone (50 mg/kg) significantly (p < 0.01) improved the survival rate infected mice up to 11 days compared with the control groups. α-pinene especially in combined atovaquone at 50 mg/kg for 2 weeks meaningfully (p < 0.05) declined oxidative stress. We found the promising in vitro anti-Toxoplasma effects of α-pinene on T. gondii RH strain. In addition, we found that α-pinene therapy particularly along with the reference drug declined the mortality rate of infected mice. Although, we just confirmed the stimulation of apoptosis and anti-inflammatory effects as the main anti-Toxoplasma mechanisms of α-pinene; however, more surveys concerning the accurate mechanisms, toxicity, and efficacy on other T. gondii strains are required to confirm these results.

Chemical Composition, Apoptotic Activity, and Antiparasitic Effects of Ferula macrecolea Essential Oil against Echinococcus granulosus Protoscoleces.

BACKGROUND: Today, the present protoscolicidals used to minimize the serious risks during hydatid cyst surgery are not completely safe and have various adverse side effects. The present study aimed to evaluate the chemical composition and apoptotic activity of Ferula macrecolea essential oil (FMEO) as well as its in vitro and ex vivo protoscolicidal effects against hydatid cyst protoscoleces. METHODS: Gas chromatography/mass spectrometry (GC/MS) analysis was performed to determine the chemical composition of FMEO. Protoscoleces of hydatid cysts were collected from liver fertile hydatid cysts of infected sheep and were then treated with various concentrations of the essential oil (75, 150, and 300 µL/mL) for 5-60 min in vitro and ex vivo. Then, by using the eosin exclusion test, the viability of the protoscoleces was studied. The caspase-3-like activity of the FMEO-treated protoscoleces was also evaluated through the colorimetric protease assay Sigma Kit based on the manufacturer's instructions. RESULTS: According to GC/MS, the main constituents of the essential oil were terpinolene (77.72%), n-nonanal (4.47%), and linalool (4.35%), respectively. In vitro, the maximum protoscolicidal activity of FMEO was observed at the concentrations of 150 and 300 µL/mL, such that 100% of the protoscoleces were killed after 30 and 20 min of exposure, respectively. Based on the obtained findings, the results demonstrate that FMEO required a longer time to kill protoscoleces ex vivo; after 12 min of exposure to FMEO, only 13.4% of the protoscoleces remained alive. After 48 h of the treatment of protoscoleces, FMEO, in a dose-dependent manner and at doses of 75, 150, and 300 µL/mL, induced the activation of the caspase enzyme by 24.3, 35.3, and 48.3%, respectively. CONCLUSIONS: Our findings demonstrate the potent protoscolicidal effects of FMEO in vitro and ex vivo; however, further studies are required to assess the safety and the efficiency of FMEO as a promising scolicidal agent in a preclinical model and clinical setting.

Myrtus communis Essential Oil; Anti-Parasitic Effects and Induction of the Innate Immune System in Mice with Toxoplasma gondii Infection.

BACKGROUND: Myrtus communis (M. communis) is a wild aromatic plant used for traditional herbal medicine that can be demonstrated in insecticidal, antioxidant, anti-inflammatory, and antimicrobial activity of its essential oils (MCEO). AIM: The present study aimed to evaluate the prophylactic effects of M. communis essential oil (MCEO) against chronic toxoplasmosis induced by the Tehran strain of Toxoplasma gondii in mice. METHODS: Gas chromatography/mass spectrometry (GC/MS) analysis was performed to determine the chemical composition of MCEO. Mice were then orally administrated with MCEO at the doses of 100, 200, and 300 mg/kg/day and also atovaquone 100 mg/kg for 21 days. On the 15th day, the mice were infected with the intraperitoneal inoculation of 20-25 tissue cysts from the Tehran strain of T. gondii. The mean numbers of brain tissue cysts and the mRNA levels of IL-12 and IFN-γ in mice of each tested group were measured. RESULTS: By GC/MS, the major constituents were α-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%), respectively. The results demonstrated that the mean number of T. gondii tissue cysts in experimental groups Ex1 (p < 0.05), Ex2 (p < 0.001) and Ex3 (p < 0.001) was meaningfully reduced in a dose-dependent manner compared with the control group (C2). The mean diameter of tissue cyst was significantly reduced in mice of the experimental groups Ex2 (p < 0.01) and Ex3 (p < 0.001). The results demonstrated that although the mRNA levels of IFN-γ and IL-12 were elevated in all mice of experimental groups, a significant increase (p < 0.001) was observed in tested groups of Ex2 and Ex3 when compared with control groups. CONCLUSION: The findings of the present study demonstrated the potent prophylactic effects of MCEO especially in the doses 200 and 300 mg/kg in mice infected with T. gondii. Although the exceptional anti-Toxoplasma effects of MCEO and other possessions, such as improved innate immunity and low toxicity are positive topics, there is, however, a need for more proof from investigations in this field.

Resistance rate and novel virulence factor determinants of Arcobacter spp., from cattle fresh meat products from Iraq.

Arcobacter spp colonize in human and animals intestine and cause food-associated infections. Hence, characterization of their virulence potential and health impacts is required. Our subject was isolation and characterization of Arcobacter spp, from meat marketplaces. A total of 1297 fresh raw cattle meat samples were purchased randomly from various marketplaces in Baghdad, Iraq. One-hundred and twenty isolates were identified, including Arcobacter butzleri (A. butzleri n = 100) and Arcobacter cryaerophilus (A. cryaerophilus n = 20). Susceptibility to antimicrobials was examined using Kirby-Bauer disc diffusion method. Molecular investigation of antibiotic resistance and virulence factors was also conducted using polymerase chain reaction (PCR) technique. Most of A. butzleri were resistant to tetracycline (72%), amoxicillin (69%), erythromycin (67%) and cefoxitin (66%), while 33% and 6% of them were resistant to ceftazidime and carbapenems, respectively. All were susceptible to gentamicin, colistin and fosfomycin. Fifty-five and nine isolates of A. butzleri and A. cryaerophilus were respectively multidrug-resistant (MDR). The existence of tetA, tetB, dfrA, sul1, blaCTX-M1 and blaIMP included 61%, 58%, 57%, 34%, 46% and 3%, respectively. The virulence genes cadF, irgA, tylA, cdtC and cdtA genes were detected in all the A. butzleri and A. cryaerophilus isolates. While, ciaB mviN and pldA genes were respectively detected in 91%, 88% and 84% of A. butzleri and 97%, 93% and 87% of A. cryaerophilus isolates. There was a significant relation between MDR and existence of virulence genes. Existence of pathogenic and drug-resistant- Arcobacter spp in raw meat is a threat for human health, necessitating confirmation of quality and safety of meat products.

Zataria Multiflora bois as an auspicious therapeutic approach against Echinococcus granulosus: Current status and future perspectives.

Hydatidosis caused by Echinococcus granulosus is a major zoonotic diseases. In addition to imposing heavy economic losses, the disease is a public health problem worldwide. The larval stage of the parasite (hydatid cyst) is formed in a wide range of domestic, wild and human beings as intermediate hosts. On the other, its recurrence has been reported anywhere as a reemerging disease. Although the cysts have some evading mechanisms, both human TH1 and TH2 cells subsets are stimulated. Because of increasing resistance and adverse effects of medications such as abnormalities of liver and other organs functions and abdominal pain, seeking alternative therapeutic approaches to be inexpensive, easy available, with low side effects and toxicity seems essential. However, the lack of information on the social and economic welfares of herbal medicines for the industrial scale application is a limitation. Zataria Multiflora bois (ZMB) has exhibited huge advantages and tremendous protoscolicidal effects as demonstrated by numerous studies and its combination therapies with anti-parasitic drugs have exerted desirable outcomes in vitro and in vivo. Noticeably, the compound confers negligible side effects or toxicity even at high concentrations. ZMB has exhibited promising inhibitory effects against hydatid cyst, particularly when combined with chemical drugs and in formulations of nanoemulsions. Its immunomodulatory effects include increase of nitric oxide production (NO) and protection of hepatic cells (Kupffer cells, fat-storing cells, and endothelial cells), enhancement of macrophages and T cells and increase of cytokines production. This study aimed at assessment of ZMB traits for application against hydatid cyst protoscolices.

Use of immunological methods to the detection of toxoplasmosis and heat shock protein HSP70 in men.

Toxoplasmosis is a contagious infection caused by the parasite Toxoplasma gondii, affecting many different body organs, and its effect varies depending on the location of the infection and the condition of the host's immune system, this study was the first experiment in Al-Qadisiyah governorate to detect toxoplasmosis and thermal shock protein in males. 200 male students between the ages of 18-23 years at University of Al-Qadisiyah were persuaded to perform the examination, the immunological method ELISA showed to presence of T. gondii in 39/200 (19.5%), (10%) positive IgG antibodies and (6%) positive for IgM, statistical analysis indicates that there are significant differences between antibody presence rates between control and infection group, due to the inaccuracy of serological tests in some cases due to the reliance on the presence of antibodies, the present study used molecular methods to examine the samples again as a step to confirmation and to reach the most accurate results, depending on the gene B1 special for the parasite by using real-time PCR technique, and the results indicated a positive proportion was 15/200 (7.5%) as modern infection, the study also assessed the level of the heat shock protein HPS70 in the positive samples of infection and showed a high concentration of HPS70 in the blood samples for infected males (28.21) ng/ml compared with control group (6.96) ng/ml, statistical analysis showed significant differences between them.

Bacterial l-asparaginases for cancer therapy: Current knowledge and future perspectives.

l-Asparaginases hydrolyzing plasma l-asparagine and l-glutamine has attracted tremendous attention in recent years owing to remarkable anticancer properties. This enzyme is efficiently used for acute lymphoblastic leukemia (ALL) and lymphosarcoma and emerged against ALL in children, neoplasia, and some other malignancies. Cancer cells reduce the expression of l-asparaginase leading to their elimination. The l-asparaginase anticancerous application approach has made incredible breakthrough in the field of modern oncology through depletion of plasma l-asparagine to inhibit the cancer cells growth; particularly among children. High level of l-asparaginase enzyme production by Escherichia coli, Erwinia species, Streptomyces, and Bacillus subtilis species is highly desirable as bacterial alternative enzyme sources for anticancer therapy. Thermal or harsh conditions stability of those from the two latter bacterial species is considerable. Some enzymes from marine bacteria have conferred stability in adverse conditions being more advantageous in cancer therapy. Several side effects exerted by l-asparaginases such as hypersensitivity should be hindered or decreased through alternative therapies or use of immune-suppressor drugs. The l-asparaginase from Erwinia species has displayed remarkable traits in children with this regard. Noticeably, Erwinia chrysanthemi l-asparaginase exhibited negligible glutaminase activity representing a promising efficiency mitigating related side effects. Application of software such as RSM would optimize conditions for higher levels of enzyme production. Additionally, genetic recombination of the encoding gene would indisputably help improving enzyme traits. Furthermore, the possibility of anticancer combination therapy using two or more l-asparaginases from various sources is plausible in future studies to achieve better therapeutic outcomes with lower side effects.