Effects of metformin on autoimmune immunoglobins and interferon-γ in patients with early diagnosed pemphigus vulgaris: a prospective clinical trial.

The management of pemphigus vulgaris (PV) is challenging. This study aimed to evaluate the immunomodulating effects of metformin on PV. The study was conducted in two phases: in the first phase, patients received routine first-line treatment (prednisolone plus azathioprine) for 2 months, then in the second phase, metformin was added to this regimen for another 2 months. After addition of metformin to the first-line medications, significant reductions were seen in serum IgG1 (reduced from 534.92 ± 134.83 mg/dL to 481.58 ± 130.46 mg/dL, P < 0.001), IgG4 (51.83 ± 27.26 mg/dL to 44.50 ± 26.05 mg/dL, P < 0.001) and interferon-γ (277.99 ± 108.71 pg/mL to 45.05 ± 17.080 pg/mL, P = 0.03) concentrations. The suppressant effect of metformin was greatest on IgG4 (coefficient of variation 1.28), the dominant subclass of IgG involved in PV. Metformin could have immunomodulating effects on PV with controlling effects on steroid complications.

Protective effect of L-carnitine on platelet apoptosis during storage of platelet concentrate.

BACKGROUND: Platelet apoptosis is considered as one of the important factors involved in platelet storage lesion (PSL) and affect the quality of platelets during storage. The beneficial effect of L-carnitine (LC) on platelet apoptosis during platelet concentrates (PCs) storage has not been fully investigated. The aim of this study was to evaluate the effects of LC on platelets of PC regarding their apoptosis markers during storage. METHODS: Ten PCs from healthy donors were investigated in this study. PCs were prepared by platelet rich plasma (PRP) method and stored at 22±2°C with gentle agitation during storage. The effects of LC (15mM) on the platelet apoptosis were assessed by analyzing different indicative presence or absence of LC. Sampling was performed to evaluate apoptosis markers during platelet storage. RESULTS: The results indicated significantly higher mitochondrial membrane potential for LC-treated platelets than the untreated on the days 2 and 5 of storage (Pday2=0.001, Pday5=0.001). Phosphatidylserine (PS) exposure significantly increased on the untreated compared with LC-treated platelets on the second and third days of storage (Pday2=0.014, Pday3=0.012). Also, active caspase 3 was lower in the LC- treated platelets than the control group on the day 5 of storage (Pday5=0.004). Cytosolic cytochrome C was so significantly lower in LC-treated compared to the untreated platelets during storage time (Pday2=0.002, Pday3=0.001, Pday5=0.001). CONCLUSION: The results of this study indicate that the use of LC as an additive solution in platelets may be useful to reduce PSL by decreasing platelet apoptosis via mitochondrial pathway and increase platelet quality during storage.

HLA-DRB1*01:01, but not HLA-DRB1:1503 or HLA-DRB1*11, is associated with decreased inhibitor risk in Iranian hemophilia A patients.

BACKGROUND/OBJECTIVE: Hemophilia A is a genetic disorder through which patients suffer from recurrent bleeding. This can be caused by a defect in human plasma coagulation factor VIII. High incidence of FVIII inhibitors in some severe hemophilia A patients after FVIII therapy is a considerable complication. Determination of good predictive factors can improve the safety of this treatment. HLA-II have been shown as a predictive element for inhibitor development. The goal of this study is to determine the association between HLA-DRB1*15:03, HLA-DRB1*11 and HLA-DRB1*01:01 alleles and FVIII inhibitors in severe hemophilia A patients in Iran. MATERIALS/METHODS: HLA-DRB1 genotyping was performed using Multiplex sequences Specific Primers (PCR-SSP) in two groups of severe hemophilia A patients comprising 51 and 50 individuals with and without FVIII inhibitors respectively. The levels of inhibitor were determined through Nijmegen-modified Bethesda assay. HLA-DRB1 allele frequencies were compared between groups by using multiple logistic regression models. RESULTS: HLA-DRB1*01:01 allele frequency was significantly higher in patients without inhibitor ORadj: 2.7 (95%CI: 1.08, 6.97; P = 0.034). There wasn't any statistically significant difference in HLA-DRB1*11 allele frequency between groups ORadj: 0.7 (95%CI: 0.27, 1.82; P = 0.47). There was no connection between HLA-DRB1*15:03 and inhibitor development ORadj: 0.94 (95%CI: 0.38, 2.35; P = 0.94). CONCLUSION: An association between HLA-DRB1*01:01 and paucity of FVIII inhibitor showed that this allele has probably a protective effect in severe hemophilia A patients in Iran. Determination of the predictive and protective alleles are beneficial in pre-treatment activities and decrease the risk of unsuccessful therapy with FVIII in each population.

Effect of Ramadan fasting on glycemic control in patients with Type 2 diabetes.

BACKGROUND: Although Muslim patients with Type 2 diabetes may be exempt from fasting during Ramadan for medical reasons, a high proportion of them fast. AIM: To investigate the association between Ramadan fasting and glycemic control in patients with Type 2 diabetes. SUBJECTS AND METHODS: A prospective cohort clinical trial was designed. Eighty-eight patients with Type 2 diabetes (45 male, 43 female, age 51±10 yr) who opted to fast for at least 10 days during the month of Ramadan were recruited. Fasting blood samples were taken at the beginning and end of Ramadan, and 1 month after Ramadan, to assess fasting blood glucose (FBG), fasting insulin, full blood count, glycated hemoglobin (HbA(1c)) and fasting lipid profile. Insulin resistance was estimated using the homeostatic model assessment. Anthropometrics and blood pressure were also measured. RESULTS: There was a significant deterioration in FBG and HbA(1c) (p=0.002 and p≤0.001, respectively) and significant improvements in HDL and LDL cholesterol and body mass index after Ramadan (p<0.001). Interestingly, HbA(1c) showed a reduction 1 month after Ramadan (9.4±2% at the end of Ramadan vs 8.4±2.5% 1 month after Ramadan; p<0.001). CONCLUSION: Results from this study showed that fasting during Ramadan deteriorated the glycemic control in Type 2 diabetes patients. This was more evident in patients using oral hypoglycemic medication than diet- controlled patients. However, Ramadan fasting had small positive effects on lipid profile and body weight.

Purification of modified mycobacterial A60 antigen by affinity chromatography and its use for rapid diagnostic tuberculosis infection.

Tuberculosis has been declared a global emergency. The mainstay for its control is the rapid and accurate identification of infected individual. Antibodies to A60, one of the macromolecular antigen complexes of mycobacteria were commonly used in the rapid detection of Mycobacterium tuberculosis. The aim of this study was to prepare specific antibodies against A60 for detection of tuberculosis infection. Specific polyclonal antibodies against A60, (A60-Ab) were prepared in rabbits using 2 boosted injections of the antigen (A60). The antibodies were purified and treated with normal oral flora to remove any non-specific and cross-reactive antibodies. These antibodies were conjugated to CNBr-activated Sepharose 4B and used to isolate subunits of A60 with more specificity for M. tuberculosis. A new affinity column was designed to prepare modified (purified) A60 antigen. Purified A60 antigen (PA60-Ag) was used to develop antibody production by Immunoaffinity chromatography. 113 patients with a confirmed diagnosis of pulmonary TB at Pasteur Institute were selected for the study. The specificity of the results was analyzed with TB-rapid test by using PA60-antibodies. TB-rapid test revealed that normal oral flora-absorbed antibodies could lead to more specific results than that of the non-absorbed antibodies. The developed, modified A60 antibodies, (PA60-Ab)-rapid test showed higher sensitivity, specificity, Positive Predictive Value (PPV), Negative Predictive Value (NPV) and overall efficiency (93.0%, 86.0%, 90.0%, 91.0%, and 90.0% respectively) for the detection of the Mycobacterium antigen. Moreover, PA60-Ag showed only two protein bands of molecular weight 45 and 66kDa in SDS-PAGE while untreated A60 showed multiple bands. Thus, our study helped in the purification of a novel and well characterized A60 antigen and good diagnostic potential for detecting tuberculosis infection.

Production and characterization of liquid-stored and lyophilized reconstituted human infusible platelet membranes.

INTRODUCTION: During recent years, the need for platelet concentrate (PC) has increased. Infusible platelet membranes (IPM) have been developed as an alternative to standard PCs, with the additional advantage of long shelf-life and increased viral safety. In this study, IPM construction and the morphological and biological features of these microvesicles were surveyed to determine their binding capacities in vitro. METHODS: Thirty-five PC units prepared by the Iranian Blood Transfusion Organization were used to produce IPM. Platelets were lysed by repeated cycles of freezing and thawing, virally inactivated with wet heat in the presence of different concentrations of sodium octanoate as a heat stabilizer, and then sonicated. IPM were separated, kept at 4°C or lyophilized, and examined for binding to collagen and von Willebrand factor (VWF). RESULTS: IPM retained the binding capacity for collagen and VWF, and the extent of VWF binding was dependent on the concentration of the heat stabilizer. Additionally, a higher binding capacity was demonstrated for liquid-stored compared with lyophilized IPM. CONCLUSION: This study revealed the potential of IPM microvesicles to mimic the binding features of platelets in vitro.

The polymorphism of human leucocyte antigen-G gene in a healthy population of Iran.

Human leucocyte antigen-G (HLA-G) is a non-classical HLA class I molecule that unlike the classical HLA, has low polymorphism. This molecule, initially, found on invasive trophoblast cells and is postulated to have mediatory role in maternal-fetal interface. So far 43 alleles of HLA-G gene have been found. Studies on alleles of HLA-G gene could be useful in understanding the genetic variants of HLA-G alleles in Iranian population. The goal of this research was to determine the polymorphism of HLA-G gene in a healthy population of Iran. Genomic DNA was isolated from the whole blood of 102 randomly selected, healthy, unrelated Iranian individuals using salting-out technique followed by polymerase chain reaction (PCR) amplification of the exons 2 and 3 of HLA-G gene. For the performance of PCR-restriction fragment length polymorphism method, the PCR products were digested with several restriction enzymes and the resulted fragments were analysed using gel electrophoresis. The obtained results indicated nine alleles of HLA-G in Iranian individuals including G*01011 (4%), G*01012 (29.86%), G*01013 (10.8%), G*01015 (1.47%), G*01017 (1.96%), G*01018 (2.45%), G*01041 (29.4%), G*01043 (1.96%) and the null allele G* 0105N (18.1%). According to this study, in the Iranian subjects the most incident alleles were G*01012 and G*01041. The results for the frequency of G*01012 showed some similarity with Caucasians (36.3%).

Association of aplastic anaemia and Fanconi's disease with HLA-DRB1 alleles.

One of the most fascinating areas of research within the field of histocompatibility at present time concerns an observation that a major human histocompatibility system, human leucocyte antigen (HLA), is deeply involved in the development of a great number of diseases. Major histocompatibility complex is the most polymorphic system in the genome of different species. Recognition of HLA alleles could be useful in transplantation and disease studies. Genetic construct of HLA DRB1 was studied in Iranian normal populations and patients with aplastic anaemia and Fanconi's disease. DNA was extracted from the whole blood of 466 normal, 35 aplastic anaemia and 10 Fanconi's individuals. Then DRB1 gene polymorphism was studied by polymerase chain reaction-sequence-specific primer method. The HLA DRB1 gene analysis showed increase of DRB1*07 in aplastic anaemia patients compared to normal population (P = 0.02). According to this study, the frequency of DRB1*07 in normal individuals was 8.3, and in aplastic anaemia patients, 15.7%. Additionally, the frequency of DRB1*04 in normal, aplastic anaemia and Fanconi's individuals was 10, 5.7 and 20%, respectively. Our results of investigation showed correlation between some HLA alleles with the studied diseases. We reported the frequency of various DR types in aplastic and Fanconi's patients. This study could imply the possible role of HLA-DRB1*07 in the incidence of aplastic anaemia. Moreover, the frequency of DRB1*04, DRB1*03 and DRB1*15 alleles showed intermediate correlation with Fanconi's anaemia.

Human antibodies from phage libraries: neutralizing activity against human immunodeficiency virus type 1 equally improved after expression as Fab and IgG in mammalian cells.

Human antibodies against HIV-1 have been sought to study neutralization events on the molecular level, and for possible use in passive immune intervention. The development of phage display techniques has opened the possibility of rapidly generating human monoclonal antibodies with desired specificities. We and others have isolated human HIV-1 neutralizing antibody fragments using this technique. Bacterial expression of isolated clones does, however, differ broadly both in expression levels and functional activity. In addition, intact IgG cannot be expressed in bacteria. By transferring the genes of isolated Fab clones to a mammalian expression system we could perform a comparison of functional activity between Fab expressed in bacterial and mammalian cells, as well as Fab and whole IgG. Fab fragments expressed in mammalian cells showed increased virus neutralizing activity compared to the same Fab clones expressed in Escherichia coli, underlining the inefficiency of procaryotic expression. No difference in HIV-1 neutralizing capacity was detected between monovalent (Fab) and divalent (whole antibody) reagents expressed in CHO cells. Thus, bivalency does not always confer improved neutralization efficacy.