Modulation of mice immune responses against Schistosoma mansoni infection with anti-schistosomiasis drugs: Role of interleukin-4 and interferon-gamma.

Objective: Schistosoma mansoni (S. mansoni) is endemic in Africa, the Middle East, South America, and the Caribbean. This study investigated the modulation of immune response against S. mansoni through estimation of interleukin-4 (IL-4) (Th2 cytokine) and interferon-gamma (INF-γ) (Th1 cytokine) under the effect of anti-schistosomal drugs. Methods: Laboratory bred female albino mice (n = 120) were divided into the following groups: untreated mice, S. mansoni infected mice, S. mansoni infected mice treated with artemisinin (ART), arachidonic acid (ARA), nifedipine or praziquantel (PZQ). Levels of IL-4 and INF-γ cytokines in the serum samples of treated and untreated mice were determined by enzyme-linked immunosorbent assay and the results were further validated by measuring the mRNA levels IL-4 and INF-γ using quantitative real-time polymerase chain reaction. Results: Anti-schistosomiasis drugs ART and ARA increased the levels of Th2 cytokine IL-4 (P < 0.05), whereas PZQ drug decreased the response of IL-4 (P < 0.05). However, nifedipine was found to be ineffective in modulating the response of IL-4 (P > 0.05). As far as Th-1 cytokine IFN γ was concerned, only PZQ increased its levels (P < 0.05), whereas other tested anti-schistosomiasis drugs; ART, ARA, and nifedipine were found to be infective (P > 0.05). Conclusions: These findings indicated that anti-schistosomiasis drugs ART, ARA, and PZQ play a role in the modulation of expression of Th2 cytokines. Whereas, only PZQ may play a role in the modulation of Th1 cytokines. These findings provide a scope for the formulation of novel anti-schistosomal drugs as well as in the therapeutic management of patients infected with S. mansoni.

Toxoplasmosis in immunocompetent Saudi women: Correlation with vitamin D.

OBJECTIVE: Toxoplasma gondii (T. gondii) is a life-threatening parasite particularly infecting the immunocompromised women. Deficiency of vitamin D is well reported in several infectious disorders. This study was undertaken to investigate a correlation of vitamin D deficiency with the onset of T. gondii infection in immunocompetent women from the central of Saudi Arabia. METHODS: Blood samples were collected from 304 Saudi women from the Qassim region of Saudi Arabia. Specific immunoassays were used to determine the levels of T. gondii immunoglobulin G and vitamin D. The SPSS and the Prism Graph Pad statistical software were used for the data analysis. RESULTS: Out of 304 women, 18.8% were found to be positive for toxoplasmosis. Interestingly, the serum levels of vitamin D in toxoplasma positive cases were found to be significantly low as compared with the levels of vitamin D in toxoplasma negative cases. Moreover, sociodemographic risk factors such as age, residence location, and consumption of fruits/vegetables were also found to be associated with vitamin D deficiency and with the seroprevalence of toxoplasmosis. CONCLUSION: This study investigated a direct correlation of vitamin D deficiency with the severity of the toxoplasmosis in Saudi women. Therefore, it is predicted that vitamin D supplementation may provide protection against toxoplasma infection.

Basil Essential Oil and Its Nanoemulsion Mitigate Non Alcoholic Steatohepatitis in Rat Model with Special Reference to Gut Microbiota.

The present research evaluated the protective effect of basil essential oil nanoemulsion (BNO) and its parent basil essential oil (BO) towards steatohepatitis. Chemical composition of BO was assessed followed by formulation into different BNOs using the low energy spontaneous emulsification technique. An ideal formula of BNO was selected among the others based on its ultra-fine particle size (15.42 nm) and physical stability at 25-37°C, which was then tested in steatohepatitis rat model along with BO. Rats were divided into four groups, the first was fed on balanced diet (C), and the other groups were maintained on high fructose saturated fat diet deficient in choline to induce steatohepatitis, one of such groups served as control steatohepatitis (SC), the other groups received daily oral dose of BO and BNO, respectively. Microbiota (Firmicutes and Bacteroidetes) were counted in colon content and their ratio (F/B) was calculated. Liver fat, plasma lipid profile, plama interlukin-6, plasma lipopolysaccharides and plasma and colon content of lipocaline were assessed with histopathological examination of liver and colon. Results showed that the major volatile components of BO were linalool (60.9 %), eugenol (5.1 %) and eucalyptol (9.5%). SC group exhibited significant increase in liver lipids, plasma triglycerides, total cholesterol (TC), low density lipoprotein cholesterol and significant reduction in high density lipoprotein-cholesterol (HDL-C) compared to C group. Significant increase in plasma TC/HDL-C, interlukin-6, and lipocaline and F/B ratio and lipocaline in colon content were demonstrated in SC group without changes in plasma lipopolysaccharides compared to C. Histopathology of SC group showed liver fatty degeneration and fibroblasts activation while the colon demonstrated erosion and mucosal epithelium detachment. Treatment with either BNO or BO showed improvement compared to SC group. BNO was superior in reducing F/B ratio, liver lipids and histopathological changes. BO was more efficient in reducing TC, triglycerides and low density lipoprotein cholesterol. It is concluded that BO and BNO reduced the progression of nonalcoholic steatohepatitis in rat model. Gut microbiota in relation to steatohepatitis and related new therapies needs further investigations.

Evaluation and characterization of some protective culture metabolites in free and nano-chitosan-loaded forms against common contaminants of Egyptian cheese.

This research attempted to inspect the contribution of lactic acid bacteria (LAB) with nanoparticle application in antimicrobial enhancement. Seven lactic acid cultures-free supernatants (CFSs) in both free and nanoparticles-loaded states were examined against seven foodborne microorganisms. Lactobacillus helveticus followed by Lactobacillus Plantarum possessed considerable antimicrobial activity. Headspace GC-MS characterization of Lactobacillus helveticus CFS identified a mixture of antimicrobial and health-promoting compounds. Minimal inhibitory concentration (MIC) values for tested Gram-positive bacteria represented 50% of that for Gram-negative bacteria, 20% and 7.35% of those for fungus and yeast respectively. Nanoparticles were prepared through chitosan-tripolyphosphate nanoparticle formation giving nanospheres from in the range from 5 to 10 nm, and narrow size distribution. CFS-loaded chitosan nanoparticles (CS-NPs) significantly enhanced the overall inhibition zone diameter, as well as, the decline in MIC values for Salmonella enterica (50%) and Penicillium chrysogenum (12.5%) was observed. Lactobacillus helveticus CFS, however, displayed lower antimicrobial activity vs. nisin and natamycin, it has both antibacterial and antifungal promising activities.

The protective potential of selected lactic acid bacteria against the most common contaminants in various types of cheese in Egypt.

Dairy products, especially cheeses have a great nutritional value and a high consumption level around the world. Considering a widespread consumption of cheeses, there is a growing concern regarding safety and microbiological quality. The current study was designed to conduct a recent evaluation of cheeses microbiological quality. Sixty cheese samples from retailing Egyptian markets were analyzed on different selective microbiological media and 64 bacteria, 35 yeasts and 8 molds were isolated. Out of 60 samples; 26.6% were contaminated with Escherichia coli, 73.3% with Staphylococcus scuiri, 3.33% with Bacillus cereus, 1.66% with Salmonella enterica, and 1.66% with Pseudomonas aeruginosa. The presence of such microorganisms in cheeses referred to the wrong management in cheese manufacturing. These organisms are significant from public health view as they have been associated with the base of human food poisoning. Promising antagonistic behavior was observed using the tested lactic acid bacteria (LAB) either single or in combinations toward the undesired isolates. Lactobacillus helveticus CNRZ 32 (Lb. helveticus) was the most potent culture; recording ≥95% reduction in undesired microbial counts.

LYMPHO-PROLIFERATIVE RESPONSES TO VARIOUS FASCIOLA HEPATICA WORM'S ANTIGENS: AN IN VITRO STUDY.

Fascioliasis is an important zoonotic disease with approximately 2-4 million people infected worldwide and a further 180 million at risk of infection. F. hepatica can survive within the bile ducts for many years through its ability to suppress the host immunity with Fasciola cathepsin L1 cysteine protease and Glutathione S transferase playing an important role. The aim of the present study is to investigate the in vitro lympho-proliferative responses of hepatic hilar lymphocytes (HLN) of infected sheep in response to different F. hepatica antigens. The suppressive effects of Fasciola excretory/secretory (ES) and tegument (TEG) and their fractions were also investigated. Our results showed that both ES and TEG had significant suppressive effects on lympho-proliferation, up to 74% and 92%, respectively. When these antigens were fractionated, fraction 3 (MW of >10000-30000) of both ES (64%) and TEG (59%) in addition to fraction 4 (MW of ≤ 10000) of TEG (38%) inherited the suppressive effects. Identification of the potential molecule(s) with such suppressive effects on lymphocytes in TEG fraction 4 could reveal vaccine candidates.

THE EFFECTS OF MIRAZID FRACTIONS ON THE VIABILITY OF SCHISTOSOMULA OF:SCHISTOSOMA MANSONI: AN IN VITRO STUDY.

Schistosma infection is an endemic disease in many developing countries including Egypt. Despite the effort to control and treat the infection with praziquantel, the development of resistance is a major health problem. Myrrh extract (Mirazid*) has been used as an alternative with conflicting results. In this study the effects of Mirazide fractions on schistosomules of S. mansoni have been tested. Using column chromatography we have fractionated the molecule to 10 fractions. Two of these fractions were more potent and cause damage to the schistosomules detected by toluidine blue dyes and confirmed by electron microscopic examination. In conclusion, identification and purification of the, active molecules in these fractions may potentiate the: antischistosomal effects of Mirazid®.

Prevalence of Intestinal Protozoa among Saudi Patients with Chronic Renal Failure: A Case-Control Study.

It has been hypothesized that chronic renal failure (CRF) predisposes patients to infection with intestinal protozoa. We tested this hypothesis with a matched case-control study to determine the prevalence of these protozoa and their diarrhea associated symptoms among 50 patients with CRF (cases) from Taif, western Saudi Arabia. Fifty diarrheal patients without CRF were recruited in the study as controls. Participants were interviewed by a structured questionnaire and stool samples were collected. Samples were thoroughly examined with microscopy and three coproantigens detection kits. Enteric protozoa were detected in 21 cases and 14 controls. Blastocystis spp. were the most predominant parasite (16% in cases versus 8% in controls), followed by Giardia duodenalis (10% in cases versus 12% in controls) and Cryptosporidium spp. (10% in cases versus 6% in controls). Cyclospora cayetanensis was identified in two cases, while Entamoeba histolytica was described in one case and one control. Intestinal parasitism was positively associated with the male gender, urban residence, and travel history. Clinical symptoms of nausea/vomiting and abdominal pain were significantly varied between the parasitized cases and controls (P value ≤ 0.05). Given the results, we recommend screening all diarrheal feces for intestinal protozoa in the study's population, particularly those with CRF.

Hypoxia-induced epigenetic modifications are associated with cardiac tissue fibrosis and the development of a myofibroblast-like phenotype.

Ischemia caused by coronary artery disease and myocardial infarction leads to aberrant ventricular remodeling and cardiac fibrosis. This occurs partly through accumulation of gene expression changes in resident fibroblasts, resulting in an overactive fibrotic phenotype. Long-term adaptation to a hypoxic insult is likely to require significant modification of chromatin structure in order to maintain the fibrotic phenotype. Epigenetic changes may play an important role in modulating hypoxia-induced fibrosis within the heart. Therefore, the aim of the study was to investigate the potential pro-fibrotic impact of hypoxia on cardiac fibroblasts and determine whether alterations in DNA methylation could play a role in this process. This study found that within human cardiac tissue, the degree of hypoxia was associated with increased expression of collagen 1 and alpha-smooth muscle actin (ASMA). In addition, human cardiac fibroblast cells exposed to prolonged 1% hypoxia resulted in a pro-fibrotic state. These hypoxia-induced pro-fibrotic changes were associated with global DNA hypermethylation and increased expression of the DNA methyltransferase (DNMT) enzymes DNMT1 and DNMT3B. Expression of these methylating enzymes was shown to be regulated by hypoxia-inducible factor (HIF)-1α. Using siRNA to block DNMT3B expression significantly reduced collagen 1 and ASMA expression. In addition, application of the DNMT inhibitor 5-aza-2'-deoxycytidine suppressed the pro-fibrotic effects of TGFß. Epigenetic modifications and changes in the epigenetic machinery identified in cardiac fibroblasts during prolonged hypoxia may contribute to the pro-fibrotic nature of the ischemic milieu. Targeting up-regulated expression of DNMTs in ischemic heart disease may prove to be a valuable therapeutic approach.

An 18FDG-PET/CT finding of a mediastinal abscess.

A 66-year-old man with left pyriform fossa squamous cell carcinoma (T2N0) was treated with chemoradiation. Nine months later, an emergency tracheostomy was performed for respiratory distress. Contrast-enhanced neck and thorax CT demonstrated a right pyriform mass. FDG-PET/CT showed linear increased uptake extending superiorly from the tracheostomy to the right Eustachian tube and inferiorly to the posterior mediastinum. Postmortem examination confirmed a 7 × 2.2 cm abscess extending from the right parapharyngeal, peritracheal, and prevertebral structures to the mediastinum.

Novel in vitro assays for the characterization of EMT in tumourigenesis.

BACKGROUND: Two novel assays quantifying Epithelial to Mesenchymal Transition (EMT) were compared to traditional motility and migration assays. TGF-beta1 treatment of AY-27 rat bladder cancer cells acted as a model of EMT in tumourigenesis. METHODS: AY-27 rat bladder cancer cells incubated with 3 ng/ml TGF-beta1 or control media for 24 or 48 h were assessed using novel and traditional assays. The Spindle Index, a novel measure of spindle phenotype, was derived from the ratio of maximum length to maximum width of cells. The area covered by cells which migrated from a fixed coverslip towards supplemented agarose was measured in a novel chemoattractant assay. Motility, migration and immunoreactivity for E-cadherin, Vimentin and cytokeratin were assessed. RESULTS: TGF-beta1 treated cells had increased "spindle" phenotype together with decreased E-cadherin, decreased Cytokeratin-18 and increased Vimentin immunoreactivity. After 48 h, the mean Spindle Index of TGF-beta1 treated cells was significantly higher than Mock (p=0.02, Bonferroni test) and there were significant differences in migration across treatment groups measured using the novel chemoattractant assay (p=0.02, Chi-square). TGF-beta1 significantly increased matrigel invasion. CONCLUSION: The Spindle Index and the novel chemoattractant assay are valuable adjunctive assays for objective characterization of EMT changes during tumourigenesis.

Epigenetic events, remodelling enzymes and their relationship to chromatin organization in prostatic intraepithelial neoplasia and prostatic adenocarcinoma.

OBJECTIVE: To explore the nuclear chromatin phenotype, overall epigenetic mechanisms, chromatin remodelling enzymes and their role as diagnostic biomarkers in prostate lesions, using high-resolution computerized quantitative digital image analysis (DIA). MATERIALS AND METHODS: A tissue microarray (TMA) was constructed using paraffin wax-embedded prostatic tissues from 78 patients, containing 30 cores of benign prostatic hyperplasia (BPH), 10 of low-grade prostatic intraepithelial neoplasia (LGPIN), 38 of prostate adenocarcinoma, 20 of BPH tissue excised at 0.6-1 mm from LGPIN lesions, and 10 of BPH prostatic tissues obtained 0.6-1 mm from high-grade PIN (HGPIN) lesions. Chromatin phenotype was assessed on haematoxylin-stained sections using high-resolution texture analysis. For quantitative immunohistochemistry, antibodies raised against acetylated histone H3 lysine 9 (AcH3K9), 5'methylcytidine (5MeC) and the chromatin remodelling ATPase ISWI (SNF2H and SNF2L) were used. The immunodensity was measured using DIA to determine the epigenetic profile of the cases. At least 60 nuclei were measured from each case. RESULTS: There were many statistically significant differences in staining intensity and nuclear distribution patterns in chromatin phenotype and immunostaining (p

Schistosomiasis vaccines.

While there are many challenges in vaccine development, none is greater than that of developing vaccines against large metazoan parasites such as schistosomes, the parasitic worms that are responsible for schistosomiasis. Initial optimism stemming from the identification of the first vaccine candidate antigens that gave protection in animals has been dashed by the failure, as yet, of any of the vaccine candidate antigens to enter Phase III clinical trials. Now, despite an improved understanding of the biology of the parasites and of the immune responses they stimulate in naturally exposed populations, the vaccine effort is stalled. The control effort has switched heavily in favour of the wider use of conventional chemotherapy with praziquantel, which is now affordable by all but the poorest countries. Disagreements among researchers in the schistosome field as to whether or not a vaccine is needed have not helped convince funding agencies that schistosomiasis vaccines, rather than drugs, should be a priority. With the schistosome genome projects at an advanced stage plus the power of the proteomics, perhaps it is still too early to call time on schistosome vaccine development.