Autosomal dominant deficiency of the interleukin-17F in recurrent aphthous stomatitis: Possible novel mutation in a new entity.

Recurrent Aphthous Stomatitis (RAS) is a common oral inflammatory disease with unknown pathogenesis. Although the immune system alterations could be involved in predisposition of individuals to oral candidiasis, precise etiologies of RAS have not been understood yet. A recent study showed that autosomal dominant IL17F deficiency could cause chronic mucocutaneous candidiasis. Considering the inflammatory nature of interleukin (IL)-17F and RAS, this study was performed to check any disease-associated mutation in a number of patients with RAS. Sixty-two Iranian individuals with RAS were investigated in this study. After DNA extraction using a phenol-chloroform method from the whole blood, amplification was accomplished by polymerase chain reaction and the products were sequenced using a 3730 ABI sequencer. The results of sequencing revealed a missense, heterozygous mutation of IL17F, converting a threonine to proline in a patient with RAS (T79P). The Poly-phen software suggested a damaging probability predicting this substitution to have a harmful effect on IL-17F protein function. This mutation was checked in fifty healthy individuals, and was not detected in any of them. This is the first study showing that a mutation in IL-17F is associated with susceptibility to RAS. However, functional studies and further studies on more patients with RAS are required to confirm such association.

sICAM-1, sVCAM-1 and sE-Selectin Levels in Type 1 Diabetes.

BACKGROUND/OBJECTIVE: This study was performed to compare soluble levels of adhesion molecules between diabetic patients and controls and to assess their possible association with long-term complications of type 1 diabetes (T1D). METHODS: Forty-eight patients with T1D and 39 healthy controls were enrolled in this study. The plasma level of adhesion molecules was measured by sandwich enzyme-linked immunosorbent assay technique. RESULTS: Higher sVCAM 1 (soluble vascular cell adhesion molecule 1) levels correlated with older age of onset of T1D. The plasma level of sICAM 1 (soluble intercellular adhesion molecule 1) was significantly increased, while sE selectin was significantly decreased in patients with T1D, compared to controls. There was no significant relationship between these plasma-level variations and the long-term complications of T1D. CONCLUSION: Although plasma levels of cell adhesion molecules are different in T1D patients and healthy controls, they might not be good candidate markers for prognosis of disease.

Altered Suppressor Function of Regulatory T Cells in Type 1 Diabetes.

BACKGROUND: Type 1 diabetes (T1D) is a T cell mediated autoimmune disease targeting the insulin-producing ß cells within pancreatic islets. Autoimmune diseases may develop as a consequence of altered balance between regulatory (Tregs) and autoreactive T cells. OBJECTIVES: To evaluate Treg cells frequency and suppressive function in the peripheral blood of newly diagnosed T1D patients in comparison with healthy controls. METHODS: Fifteen new cases of T1D patients and 15 age- and sex- matched healthy controls were recruited to this study. Their peripheral blood mononuclear cells (PBMCs) were isolated and CD4+CD25+FoxP3+CD127-/low Treg cells were studied by flowcytometry technique. Thereafter, Tregs were isolated by Magnetic-Activated Cell Separation (MACS) technology and by using CFSE (carboxyfluorescein succinimidyl ester) dilution assay, their suppressive activity was evaluated in the coculture of CD4+CD25- T responder cells with Treg cells. RESULTS: The percentage of CD4+CD25+FoxP3+CD127-/low Tregs did not differ between T1D patients and healthy controls but the MFI (mean fluorescence intensity) of transcription factor FoxP3 (forkhead box protein P3) was significantly decreased in T1D patients (20.03 ± 1.4 vs. 31.33 ± 2.95, p=0.0017). Moreover, the suppressive function of CD4+CD25+CD127-/low Treg cells was significantly diminished in T1D patients in comparison with control group (35.16 ± 4.93% vs. 60.45 ± 5.26%, respectively, p=0.0015). CONCLUSION: Present study indicates an impaired immune regulation among T1D patients, characterized by defects in suppressive function and expression of FoxP3 in Treg cells without any significant decrease in their frequency in peripheral blood.

Association between a PD-1 gene polymorphism and antisperm antibody-related infertility in Iranian men.

OBJECTIVE: Programmed cell death-1 (PD-1, Pdcd1), an immunoreceptor belonging to the CD28/CTLA-4 family negatively regulates antigen receptor signalling by recruiting protein tyrosine phosphatase, SHP-2 upon interacting with either of two ligands, PD-L1 or PD-L2. This study investigates PD-1 gene polymorphism in patients with antisperm antibody-related infertility METHODS: Genotyping was performed by polymerase chain reaction and restriction enzyme digestion (PCR-RFLP), this polymorphism was genotyped in 145 Iranian subjects (61 patients with antisperm antibody-related infertility and 84 healthy controls). RESULTS: Patients frequencies of the G/A genotype in comparison with healthy controls (38.2 % vs. 32.7 %, OR =1.21, P = 0.35) were not significantly different. However, G/G and A/A genotype frequencies between patients and healthy controls were significantly different (P = 0.042, P = 0.00001, respectively). Also, allele frequencies of this polymorphism were significantly different (P = 0.0012) in patients compared to healthy controls. CONCLUSION: According to these results, there is a correlation between PD-1 gene polymorphism and susceptibility to antisperm antibody-related infertility in our study group.

Anti-cyclic citrullinated peptide antibody and rheumatoid factor isotypes in Iranian patients with rheumatoid arthritis: evaluation of clinical value and association with disease activity.

In this study we determined the frequency, sensitivity and specificity of anti cyclic citrullinated peptides (anti-CCP) IgG antibody, total rheumatoid factor (RF-T), and RF isotypes in Iranian patients with rheumatoid arthritis (RA) and their association with age, clinical and serological parameters. Anti-CCP and RF-T and RF isotypes level were measured in 418 patients and 399 healthy controls by enzyme-linked immunosurbant assay (ELISA). Additionally, serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), visual analog scale (VAS) and disease activity score (DAS28) were evaluated in RA patients. The anti-CCP was positive in 53.1% of RA patients and 4.7% of controls. The frequency of RF-T was 61.87% and 17.66% in RA patients and controls respectively. The prevalence of RF isotypes in RA patients was 46.52% for RF-IgM, 23.47% for RF-IgA and 21.74% for RF-IgG. 31.39% of RA patients were RF-IgM positive without RF-IgA and RF-IgG and 21.9% were positive for all three RF classes. The anti-CCP positive patients showed increased number of swollen joints. On the other hand, RF-T positive patients exhibited a longer disease duration, lower age of onset and also higher ESR, CRP level and increased swollen joints. RF-T titer was significantly higher in RA patients with active disease compared to remission, low and moderate active groups. The sensitivity and specificity were 53.1, 95.3 for anti-CCP antibody and 61.8, 82.3 for RF-T. Our results support that anti-CCP and RF titer maybe valuable in estimation of disease activity and other inflammatory parameters in RA patients.

Tegumental histological effects of Mirazid(®) and myrrh volatile oil on adult Fasciola gigantica.

OBJECTIVE: To evaluated the histological changes within the tegument of adult Fasciola gigantica (F. gigantica) that led to the gross changes that were visible externally. METHODS: The effects of oleoresin extract of myrrh (Mirazid(®)), myrrh volatile oil and triclabendazole sulphoxide (reference drug) on the tegumental structure of adult F. gigantica following treatment in vitro had been determined by light microscopy. RESULTS: The internal changes in the tegument observed in this study were compatible with surface changes seen in the previous scanning electron microscopy study, using the same drugs. The swelling of tegumental syncytium was a particular feature of their action, but its level was much greater with myrrh volatile oil, in which vacuolization of the tegument and loss of spines were observed. CONCLUSIONS: The present study demonstrated the fasciocidal properties of Mirazid(®) oleoresin extract, and it might be possible to reinforce its fasciocidal activity by increasing its content of myrrh volatile oil.

GM-CSF differentially regulates eosinophil and neutrophil adhesive interactions with vascular endothelium in vivo.

Allergic airway inflammation is characterized by elaboration of cytokines and chemokines leading to recruitment of inflammatory leukocytes, predominantly eosinophils, to the airways. Granulocyte macrophage colony stimulating factor (GM-CSF) is generated in the lungs of human subjects with asthma in response to allergen challenge and is necessary for the development of allergen-induced bronchial eosinophilia in mice. The effect of GM-CSF on human eosinophil and neutrophil interactions with the vascular endothelium under conditions of blood flow was investigated in post-capillary venules of the rabbit mesentery by intravital microscopy.While GM-CSF significantly reduced the rolling fraction of neutrophils in vivo and induced consistent shedding of neutrophil L-selectin in vitro, its effect on eosinophil rolling was variable. Eosinophils from 57% of the donors demonstrated inhibition of rolling, while eosinophils from the remaining 43% of donors demonstrated no inhibition or increased rolling. The variable effect of GM-CSF on inhibition of eosinophil rolling was associated with variable shedding of L-selectin in vitro. In contrast to the differential effect of GM-CSF on neutrophils versus eosinophils, stimulation with phorbol myristate acetate demonstrated a similar degree of inhibition of rolling and L-selectin shedding by neutrophils and eosinophils suggesting that there was no defect in L-selectin shedding in the eosinophil donors who did not respond to GM-CSF. Overall, these studies demonstrate that GM-CSF consistently inhibits interaction of neutrophils with endothelium in vivo, whereas its effect on eosinophil-endothelial interactions is variable. GM-CSF may thus be one factor accounting for the varying percentage of eosinophils and neutrophils recruited to sites of allergic inflammation in different individuals.

Mirazid in treatment of three zoonotic trematodes in Beni-Sweif and Dakhalia Governorates.

A total of 60 patients with schistosomiasis (40), fascialiosis (15) and heterophyiasis (5) were selected Beni-Sweif and Mansoura Districts and subjected to history taking, clinical examination, Kato thick smear, sedimentation and hatching test (for schistosomiasis cases) at the beginning of the study, 2 & 3 months after treatment with Oleo-resin of Myrrh (Mirazid) in a dose of 10 mg/kg/day for 6 consecutive days an hour before breakfast. The results showed a significant improvement in symptoms with minimal negligible or no side effects. The cure rates, 2 & 3 months after treatment were 80.7%% & 11.8%% for schistosomiasis, 93.3% & 6.6% for fascioliasis, and 100% for heterophyiasis. The clinical picture of schistosomiasis before treatments were easy fatigability, anorexia, nausea, vomiting, epigastria pain, abdominal distention, right upper guardant pain, colicky abdominal pain, left upper and/or lower guardant pain, abdominal rumbling, dysentery, diarrhea, rectal bleeding, constipation, and alternating bowel habit. Those of fascioliasis were abdominal distention, dripping of saliva, right upper guardant, colicky abdominal pain, weight loss, easy fatigability, intermittent jaundice, anorexia, nausea, vomiting, epigastria, left upper and/or lower quadrant pain, right layer quadrant pain, loin pain, abdominal rumbling, diarrhea, constipation, and alternating bowel habit. The safety and efficacy of C. molmol extract in treating heterophyiasis (100%), fascioliasis (100%) and schistosomiasis (92.5%) were documented.

Association of CTLA4 gene polymorphism in Iranian patients with ankylosing spondylitis.

INTRODUCTION: Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is a costimulatory molecule expressed by activated T cells. This study was performed to investigate the allele and genotype frequencies of CTLA4 gene polymorphisms in Iranian patients with ankylosing spondylitis (AS). METHODS: One hundred and fifty-seven patients with AS and 103 controls were included in this study. Polymorphisms of CTLA4 gene at positions +49 (in exon 1), -318, and -1,147 (in the promoter region) were studied on the genomic DNA using PCR restriction fragment-length polymorphism method. RESULTS: The frequencies of the T allele at position -1147 in the patients with AS was significantly increased in comparison with the control group (11% vs. 5%, P = 0.004); whereas the frequencies of C allele at the same position were significantly decreased in the patient group (89% vs. 95%, P = 0.004). Comparison of genotype frequencies at this position showed that the frequency of CT genotype in comparison with other genotypes was overrepresented in the patient group (20% vs. 8%, P = 0.012), while the CC genotype in comparison with other genotypes was decreased (79% vs. 91%, P = 0.012). There was no significant difference on frequencies of genotypes at the positions -318 and +49. CONCLUSION: This study could suggest an association between specific allele in the promoter region of CTLA4 gene and AS disease.

Increased expression of TRAIL and its receptors on peripheral T-cells in type 1 diabetic patients.

BACKGROUND: Type-I diabetes is an autoimmune inflammatory disease in which pancreatic beta-cells are selectively destroyed by infiltrating cells. TNF-related apoptosis-inducing ligand (TRAIL) is a type-II membrane protein of the TNF superfamily which is expressed in different tissues, including pancreas and lymphocytes. In humans, TRAIL interacts with four membrane receptors. TRAIL-R1 and TRAIL-R2 have cytoplasmic death domains, and can activate both caspases and NFkappaB pathways. The other two receptors, TRAIL-R3 and TRAIL-R4, are decoy receptors not capable of activating caspase cascade but may activate NF-kappaB and block apoptosis. As human beta cells are sensitive to TRAIL induced apoptosis, signaling via these molecules is considered to be a probable way of beta cell destruction. These molecules also are important in suppression of autorective T cells and immunoregulation. OBJECTIVE: To explore the importance of TRAIL and its receptors at pathogenesis of type-I diabetes, we compared expression of these molecules on T-cells of diabetic patients and healthy controls. METHODS: In this study, expression of TRAIL and its receptors at protein and mRNA levels were studied in freshly isolated peripheral T cells of 55 type I diabetic patients and 50 healthy individuals by flowcytometry, western blot and RT-PCR. RESULTS: We found that expression of TRAIL and its receptors in peripheral T-cells at both protein and mRNA levels are significantly increased in patients (except for TRAIL-R2 mRNA which was slightly higher in controls) but increase in TRAIL, TRAIL-R3 (2.7% vs. >0.5%) and TRAIL-R4 (2.6% vs. >0.5%) is more considerable. sTRAIL in sera of patients was significantly lower than in controls (P=0.01). CONCLUSION: Our results explain resistance of autoreactive T-cells to immunoregulatory mechanisms. Besides, increased expression of TRAIL in autoreactive T-cells may play an important role in beta-cell destruction. Lower level of sTRAIL in diabetic patients may be a reason for hyperactivation of autoreactive T-cells.

Mirazid (Commiphora molmol) in treatment of human heterophyiasis.

A total of 3278 patients attended Mansoura University Hospitals' Clinics with gastro-intestinal troubles suggesting parasitosis were examined by direct smear and by Kato-Katz methods for parasites especially Heterophyes heterophyes. Fifty clinically and parasitologically proved pure heterophyiasis patients were given Mirazid as two capsules for 9 successive days on an empty stomach an hour before breakfast. All the cases were subjected to history taking and clinical examination before treatment and were followed-up for four weeks post-treatment. There was an overt clinical and parasitological improvement. A total of 47 out of 50 (94%) were cured. Another course was given to the three patients who were still positive, but only two of them were cured (66.7%). The overall cure rate was 49/50 (98%) and none had any side effect. The history and treatment of this zoonotic parasite was critically discussed.

Commiphora molmol extracts as plant molluscicide against Lymnaea natalensis.

Two extracts from the herbal plant, Commiphora molmol showed a high molluscicidal effect against Lymnaea natalensis. The oil extract was more potent than the oleo-resin. A concentration of 10 ppm of the oil extract killed 100% of L. natalensis after 5th day, but the oleo-resin extract killed 100% of them at a concentration 20 ppm after 5th day.

Association of the Phe206Leu allele of the L-selectin gene with coronary artery disease.

BACKGROUND AND AIMS: The aim of this study was to assess the association between the L-selectin Phe206Leu polymorphism and coronary artery disease. METHODS: A total of 322 patients (221 men and 101 women) with coronary artery disease in one or more vessels documented by angiography were studied; 157 subjects (85 men and 72 women) without atherosclerosis were included as controls. All subjects were genotyped for the L-selectin Phe206Leu gene polymorphism using polymerase chain reaction with sequence-specific primer (PCR-SSP). To assess disease severity, all patients were classified by numbers of coronary arteries with 50% stenosis. RESULTS: A significantly increased frequency of the 206Leu mutant allele was observed in patients with coronary artery disease compared to the controls. The 206Leu allele frequency occurred in 42% of the patients with coronary artery disease compared to 30% of the controls (p<0.009). No association was found between the severity of coronary artery disease and the L-selectin Phe206Leu polymorphism. CONCLUSION: Our findings suggest that carriage of L-selectin 206Leu mutant allele could contribute to susceptibility of Iranian individuals to contracting coronary artery disease.

Effect of Mirazid (Commiphora molmol) on experimental heterophyidiasis.

Mirazid (MZ), an oleo-resin extract derived from Myrrh was reported in several experimental and clinical trails to be safe and effective against other trematodes like schistosomiasis and fascioliasis. This experimental work aimed at investigating the possible efficacy of MZ against heterophyids (Pygidiopsis genata), using praziquantel as a therapeutic control. Results showed that MZ in emulsion form is a promising drug for the treatment of heterophyidiasis, as proved by significant reduction of worm count, overt surface tegumental changes like deformity and erosion of tegumental spines observed by scanning electron microscope (SEM). The effective dose regimen was 500 mg/kg/d for 3 successive days, produced 100% reduction in worm load. The proved efficacy of the drug, together with reported low toxicity, relative to praziquantel, favours its use as a natural new alternative therapy for the treatment of human heterophyidiasis.

Compatibility of Biomphalaria alexandrina snails to infection with Schistosoma mansoni after exposure to sublethal concentrations of Myrrh.

The effect of exposing B. alexandrina to sub-lethal dose (LC10 & LC20) of Myrrh, on its susceptibility to infection with S. mansoni miracidia were determined. Starting three weeks post miracidial exposure, cercarial shedding was monitored. No shedding of cercariae were observed from snails treated with LC20. In snails treated with LC10, longer prepatent cercarial and shorter cercarial production periods than those of control group were recorded. The number of infected snails and of shedding cercariae were decreased. The study revealed that sublethal values of myrrh decreased the compatibility of B. alexandrina to S. mansoni infection thus playing an important role in the control of schistosomiasis.

Circulating ICAM-1, VCAM-1, E-selectin, P-selectin, and TNFRII in patients with coronary artery disease.

OBJECTIVE: To evaluate the relationship between the serum concentration of tumor necrosis factor receptor 2 (TNFRII) and some adhesion molecules [including intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), P-Selectin, and E-Selectin] and coronary artery stenosis. DESIGN AND SETTING: Observational (cross-sectional) study in a University Heart Hospital in Tehran, Iran. PATIENTS: 75 patients with angiographically proven coronary artery disease were compared with 81 individuals who had undergone coronary angiography with no significant evidence of stenosis (control subjects). METHODS: Soluble adhesion molecules and TNFRII were determined by enzyme-linked immunosorbent assay technique. sICAM-1 and sP-selectin values were significantly higher in patients with coronary artery disease than in control subjects [146(38) vs. 132(48) p < 0.04 and 275(107) vs. 241(104) ng/ml p < 0.04 respectively]. Multiple logistic regression analysis showed sICAM-1 an independent discriminating risk factor for coronary artery disease (p < 0.03). Prediction models that incorporated sICAM-1 in addition to other established coronary risk factors were significantly better at predicting risk than the models based on the other risk factors alone. Multiple regression analysis indicated that sP-selectin levels were greater in patients with single-vessel disease than in the respective normal (p < 0.01). CONCLUSIONS: Our findings suggest that sICAM-1 has an association with s1 coronary artery disease as such; the evaluation of this marker may improve the coronary risk assessment in Iranian patients.