Colonic mucosal eosinophilia and immunohistochemical expression of COX-2 and NF-kB in patients with irritable bowel syndrome.

BACKGROUND: There is growing evidence that eosinophilic infiltration can release mediators which are harmful to the intestinal epithelium in patients with irritable bowel syndrome (IBS). Although cyclooxygenase 2 (COX-2) and nuclear factor-kappa beta (NF-kB) expression had been previously reported to increase in many inflammatory conditions, there is a paucity in data investigating their expressions in IBS. Our aim was to evaluate colonic mucosal eosinophilia and immunohistochemical expression of COX-2 and NF-kB in patients with irritable bowel syndrome. METHODS: A total of 80 patients who met the inclusion criteria of IBS based on Rome IV symptoms questionnaire were subjected to abdominal ultrasound, laboratory investigations, serum immunoglobulin E (IgE) level assessment and colonoscopic examination. Immunohistochemistry was performed to detect COX-2 and NF-kB expression in colonic biopsies obtained from IBS patients. RESULTS: Histopathological examination showed that 60 colonic biopsy specimens (75%) showed few mixed inflammatory cells ≤3 cells/ HPF, 12 biopsy specimens (15%) showed eosinophilic infiltration ≥25 eosinophils/HPF and 8 biopsy specimens (10%) showed severe lymphocytic infiltration and aggregation. Colonic eosinophilic infiltrate was significantly higher among patients presented with IBS-D subtype. Serum IgE was significantly higher among patients with colonic eosinophilic infiltrate than the others. In IBS-D patients, colonic mucosa showed positive expression of COX-2 and NF-kB in 52.1% and 81.25% of cases, respectively. CONCLUSION: Patients with IBS -particularly IBS-D subtype- should undergo colonoscopy and biopsy to exclude underlying inflammatory pathology. Moreover, patients with positive COX-2 and NF-kB need further evaluation and follow-up.

Effects of iron chelating agent on Schistosoma mansoni infected murine model.

Schistosomiasis is one of the major health problems in many tropical and developing countries. Infection takes place once cerceriae penetrate human skin, then it changed into schistosomules. The schistosomules takes iron in the form of heme from host's haemoglobin, ferritin and transferrin. Iron is a vital element not only for growth and sexual maturity of schistosomules to adults but also for oogenesis. Since the trapped eggs are the pathological causative agent for most of pathogenesis and complications, the current work was designed to study the effects of early deprivation of schistosomules from iron in the host (in vivo) by chelating it with deferoxamine (DFO). The iron chelation has effects on growth, maturity and egg deposition, as well as it has ameliorative effects on liver pathology such as hepatic fibrosis. Mice were classified into four groups, normal control, DFO treated only, Schistosoma mansoni (S. mansoni) infected DFO untreated and S. mansoni infected DFO treated. The infected DFO treated mice showed significant reduction in fecal egg excretion with increased percentage of dead eggs and this was accompanied with a significant reduction of both total worm burden and hepatic egg load and increased dead egg percentage compared to the infected DFO untreated group. There was also a significant reduction in both serum and hepatic tissue ferritin concentrations in the infected DFO treated mice in comparison to the infected DFO untreated group. Additionally, a significant decrease in number and size of granulomas with subsequent improvement of liver fibrosis was recorded in the infected DFO treated group. This immunopathology was also associated with significant up regulation of Interlukine12 (IL12), Interferon gamma (IFN γ) and significant down regulation in interleukin4 (IL4), interleukin10 (IL10) in both serum and hepatic tissue in the infected DFO treated compared to other groups. Entirely, DFO succeeded in diminishing the growth, maturity and fecundity of S. mansoni with a subsequent improvement of hepatic pathology. As a result of the above findings, it can be concluded that DFO could be considered as a useful treatment against schistosomal infection.

Immunopathological assessments of human Blastocystis spp. in experimentally infected immunocompetent and immunosuppresed mice.

Blastocystis spp., one of the most common parasites colonizing the human intestine, is an extracellular, luminal protozoan with controversial pathogenesis. The host's immune response against Blastocystis spp. infection has also not been defined yet. Therefore, this research aimed to assess the potential pathogenicity of this parasite and its ability to modulate the immune response in experimental infected immunocompetent and immunosuppresed mice. These results demonstrated that the infected immunosuppressed mice were more affected than infected immunocompetent mice. Histopathological examination of the small intestine in the infected immunosuppressed mice showed that Blastocystis spp. infiltrated all the layers. Moreover, the epithelia showed exfoliation and inflammatory cell infiltration in submucosa compared to that of the infected immunocompetent mice. As well, examination of the large intestine of the infected immunosuppressed group showed severe goblet cell hyperplasia. Blastocystis spp. infiltrated all the large intestine layers compared to that of the infected immunocompetent group. Furthermore, there was a significant upregulation of the expression of proinflammatory cytokines: interleukin 12 (IL-12) and tumor necrosis factor alpha (TNF-α) in the infected immunosuppressed mice compared to that of the infected immunocompetent ones (p ≤ 0.004 and p ≤ 0.002, respectively). However, the expression of anti-inflammatory cytokines (IL-4 and IL-10) was significantly downregulated in the infected immunosuppressed group compared to that of the infected immunocompetent group one at 10 days postinfection (p ≤ 0.002 and p ≤ 0.001, respectively). The results of this study revealed that Blastocystis spp. affected the production of pro- and anti-inflammatory cytokines in both groups of mice compared to healthy normal (naive) group. Additionally, these data showed that there was a significant upregulation (p ≤ 0.005) of the locally synthesized antibody: secretary IgA (sIgA) in the gut of the infected immunocompetent mice when compared to that of the infected immunosuppressed ones.

Human wound myiasis caused by Phormia regina and Sarcophaga haemorrhoidalis in Minia Governorate, Egypt.

Myiasis is the parasitic infestation of human by the larvae (maggots) of dipterous fly that grow within the host while feeding on its tissue. Cutaneous myiasis is the most considerably encountered clinical form. Moreover, wound (traumatic) myiasis is the main clinical manifestation of cutaneous myiasis. In this research, we aimed to study the type of infesting larvae that are responsible for wound myiasis in the patients in Minia city, Egypt. Three cases of wound myiasis have been noticed among 280 patients with wounds at different parts of bodies. Two of them were diabetic patients. The third one had a history of hypertension with right side hemiplegia 2 years ago. All of them were elderly. The larvae removed from cases 1 and 3 were identified macroscopically and microscopically as the third-stage larvae of Sarcophaga haemorrhoidalis. The larvae removed from case 2 were the third-stage larvae of Phormia regina, which is very rare worldwide. In addition to the open and obsolete wound, diabetes mellitus and low socio-economic circumstances were shown to be attributed as important predisposing risk factors that led to the occurrence of myiasis in these patients.

In vivo antiprotozoan effects of garlic (Allium sativum) and ginger (Zingiber officinale) extracts on experimentally infected mice with Blastocystis spp.

Controversy surrounding the pathogenic role of Blastocystis spp. in humans and lack of well-established diagnostic criteria led to debates concerning the treatment for that organism. Furthermore, some strains develop resistance against the recommended drugs. Thus, using natural medicine has many positive aspects to address these points. In an earlier study, we addressed in vitro effect of garlic and ginger on Blastocystis spp. isolates as an alternative treatment. Accordingly, this study was conducted to evaluate in vivo activities of these two herbs on mice infected with Blastocystis spp. Antiprotozoan activities were determined by monitoring Blastocystis shedding in stools and histopathological changes of the intestine of infected mice. Additionally, assessment of the antioxidant effect (via measuring the level of malondialdehyde (MDA) production) of these herbs on the treated groups of mice was done. Also, their effects on nitric oxide (NO) production were assessed. In this work, treatment of infected mice with garlic, ginger, and nitazoxanide (NTZ) reduced the shedding of cysts significantly compared to the infected untreated group, P value ≤0.001, 0.0001, and 0.0003, respectively. As well, histopathological examination revealed that Blastocystis was frequently observed within the lumen, at the tip of the epithelium, and/ or infiltrated in an enterocyte in the infected group without treatment compared to that of the infected treated ones. Furthermore, mice infected with Blastocystis exhibited increased levels of NO (440.09 ± 3.7 vs. 276.66 ± 0.8, P ≤ 0.001) and MDA production (106.19 ± 0.43 vs. 63.06 ± 0.45, P ≤ 0.0004) compared to that of the uninfected controls. Treatment of infected mice with garlic, ginger, and NTZ reduced NO levels to 54.41 ± 1.2, 47.70 ± 1.2, and 37.43 ± 0.98 and MDA levels to 22.38 ± 0.17, 63.34 ± 3.89, and 66.76 ± 9.1, respectively. We conclude that using ginger and garlic for treatment of blastocystosis is beneficial.

In vitro effect of some Egyptian herbal extracts against Blastocystis hominis.

Blastocystis hominis is an enteric parasite that inhabits the gastrointestinal tract of humans and many animals. This emerging parasite has a worldwide distribution. It is often identified as the most common eukaryotic organism reported in human fecal samples that showed a dramatic increase in recent years. Metronidazole is the main therapy for blastocystosis. However, frequent reports of treatment failure suggesting isolates resistance to metronidazole. This study determined the growth pattern and in vitro susceptibility of B. hominis to nitazoxanide (NTZ), garlic, ginger, onion and turmeric. Fecal samples positive for Blastocystis were collected from patients with irritable bowel syndrome (IBS), and processed for culture. Cultured samples were subjected to examination by light microscopy. Herbs' extracts was freshly prepared. Drug susceptibility assays was done using 0.1 mg/ml of NTZ, garlic, ginger, onion and turmeric. Effects assessed on parasite culture after 24 hr. and 48 hr. Cultured fecal samples of B. hominis have identified several forms of the organism; vacuolar, granular, amoeboid and cyst forms within 24 hr. Nitazoxanide treatment significantly (P < 0.001) lowered the parasite number after 48 hr. (mean, 337.5 ± 17.67) /ml. The reduction rate after 48 hr. compared to PBS was 93.33%. Ginger treatment significantly (P < 0.002) lowered the number of the parasite after 48 hr. (mean, 335 ± 7.07)/ml. Moreover, garlic treatment also significantly (P < 0.002) lowered the number of the parasite after 48 hr. (mean, 382.5 ± 10.60)/ml. The reduction rates after 48 hr. in these treated samples compared to PBS were 92.98% and 92.44% respectively. However, onion, and turmeric treatments insignificantly lowered the number of the parasite after 48 hr. (P < 0.15 & < 0.22 respectively).

Some studies on spontaneous Hymenolepis diminuta infection in laboratory rats.

Hymenolepis diminuta is a tapeworm that occurs worldwide. It is known to be found commonly in areas where large amounts of food grains or other dry feed products, which are the favorite foods for rats. Transmission of disease to human is uncommon; however, it may be a serious threat for population who are living in rural areas which are suffering from excessive rodents. Here, this study had done on spontaneous H. diminuta infection in laboratory rats as a model. Out of thirty five adult laboratory rats investigated for parasitic diseases only nine (25.71%) were diagnosed positive for spontaneous H. diminuta infection. Four of them (44.44%) were found losing of weight and lacking of motility, while the others were normal. On microscopic examination, H. diminuta eggs had been found in their stool. On autopsy, small intestines were found to contain from 5-6 multi-segmented tapeworms in each rat. Histopathologically, intestinal lumen showed varying sections of H. diminuta segments with serrated borders. H. diminuta infection caused multiple mucosal ulcers with absence of intestinal villi from the surface epithelium and excessive mucin. Moreover, inflammatory cells infiltration in the connective tissue core of the villi. Furthermore, the Toluidine blue stain showed that there are Mastiocytosis. Additionally, there were goblet cells hyperplasia on using PAS. Moreover, there were high expression of cyclooxygenase 2 (COX-2), tumor necrosis factor-α (TNF-α) and inducible Nitric-Oxide Synthase (iNOs). This implicate, strong correlation between COX-2, TNF-α and iNOs expression and inflammation induced by H. diminuta.

Human Cystic Echinococcosis in the Nalut District of Western Libya: A Clinico-epidemiological Study.

Human cystic echinococcosis (CE) is an endemic disease in the Mediterranean area that has not yet been fully documented in western Libya. The present study describes the clinico-epidemiologic profile of CE in western Libya's Nalut district. From April 2008 to July 2011, 36 cases of CE were confirmed following surgical removal of cysts. The cysts were most frequently found in the liver (61.1%), followed by the lungs (19.4%), kidneys (11.1%), peritoneal cavity (11.1%), and spleen (5.6%). Among the 36 patients, 6 possessed plural cysts and 3 had cysts in 2 organs. Blood samples from this group were examined for the presence of serum anti-hydatid IgG antibodies, which revealed positivity in 25 patients (69.4%). An additional 300 blood samples were collected randomly from the inpatient and outpatient clinics at Nalut Hospital. Twenty-seven samples (9%) were found to be positive for the anti-hydatid IgG antibody among which the prevalence of infection tended to be higher in men (12%) than in women (6%). This study demonstrates that CE is a major parasitic infectious disease of public health significance in Libya, notably in the western part of the country, and that disease awareness needs to be raised nationwide.

Present status of protozoan pathogens causing water-borne disease in northern part of El-Minia Governorate, Egypt.

Little is known about the role of different water supplies in the diversity and public health significance of pathogenic protozoan parasites. Most of these organisms have been ubiquitous in waters worldwide. The numbers of waterborne infections indicate a significant risk for their transmission even by drinking water. Hence, a total of 336 water samples were collected during 2009-2013 from different water sources from different areas of northern part of El-Minia Govemnorate, Egypt and were investigated for pathogenic protozoa. They were examined by direct microscopy followed by Modified Ziehl- Neelsen and Giemsa stains. 140 samples (41.7 %) were positive (statistically significant; P value P<0.0003). Prevalence rates were in Summer (66.7%/), Spring (51.1%), Fall (26.2 %) and Winter (22.6%). These data were statistically significant (P<0.0001). The commonest protozoa detected as a single infection was Cryptosporidium sp. (53.17%) found in all water supplies, followed by Blastocystis hominis (15.87%), Cyclospora caytenensis (11.9%), Entamoeba hiseolytica/dispar (8.73%) Giardia lamblia (6.35%() and Naegleria sp., (3.97%). Moreover, there were 14 samples with mixed parsitic infection: they were Cryptosporidium sp. and B. hominis in six samples, Cryptosporidum sp. and C. caytenenisin five samples and Cryptosporidium sp. and E histolytica/dispar in three samples. The most common contamninated water source was ponds where 32 samples (66.7%) were-positive followed by canal water 30 samples (62.5%). The results were discussed and the recommendations were offered.

Therapeutic effect of alpha lipoic acid combined with praziquantel on liver fibrosis induced by Schistosoma mansoni challenged mice.

Schistosomiasis is an endemic disease in 74 countries causing more than 250,000 deaths every year. Accordingly, the development of an effective drug for eradication of schistosomiasis is an open research field. The current chemotherapy for control is praziquantel (PZQ). However, PZQ does not improve liver fibrosis. Therefore, the aim of this study is to evaluate the combined effect of alpha lipoic acid (ALA) with PZQ on the liver fibrosis induced by Schistosoma mansoni challenged mice. Evaluation was based on the worm burden count, ova load, granuloma size, and histopathology of the liver. Reduced glutathione (GSH) was measured in the tissue as a biomarker for impaired antioxidant function. Malondialdehyde (MDA) was also measured in the tissue as a biomarker for oxidative stress. The serum level of matrix metalloproteinase 1 was measured as a biomarker for fibrotic status of the liver. Liver function enzymes such as ALT, AST, and GGT were also measured. Four groups of ten mice each were used in this study. The first group was infected with 50 ± 10 S. mansoni cercariae. The second group was also infected and was treated with PZQ 9 weeks post-infection (PI). The third group was treated with PZQ and ALA 9 weeks PI. The fourth group was used a healthy control. The present study revealed remarkable improvement in all parameters measured (parasitological and biochemical) as well as significant improvement of hepatic pathology in the third group which was treated with PZQ and ALA. The treatment of mice with PZQ and ALA results in reduction in the worm burden, egg count, and granuloma size. Furthermore, this combined treatment increased the tissue level of the antioxidant (GSH) and decreased the tissue level of MDA in this group.

Opportunistic parasites among immunosuppressed children in Minia District, Egypt.

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children (P<0.0001). The most common protozoan infection in immunosuppressed group was Cryptosporidium parvum (60.2%), followed by Blastocystis hominis (12.1%), Isospora belli (9.7%), and Cyclospora caytenensis (7.8%). On the other hand, Entamoeba histolytica (24.6%) and Giardia lamblia (17.6%) were more common than other protozoans in immunocompetent children.

Gastrointestinal myiasis by larvae of Sarcophaga sp. and Oestrus sp. in Egypt: report of cases, and endoscopical and morphological studies.

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and us-ing a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.