The HLA-G 14-bp Insertion/ Deletion Polymorphism in Recurrent Spontaneous Abortion among Iranian Women

HLA-G is a non-classical HLA class Ib molecule with limited protein variability generated by  alternative  splicing.  HLA-G   displays  immunotolerant   properties  and  hence  plays important roles in the maintenance of a successful pregnancy and maternal tolerance of the semiallogenic fetus. Polymorphism of the HLA-G gene may potentially affect the biological properties of the protein, and a 14-bp insertion/deletion polymorphism in exon 8 of the 3′ untranslated region (3′ UTR) of the HLA-G gene is thought to influence HLA-G expression. To study the association of the 14-bp insertion/deletion (INDEL)  polymorphism with the risk of recurrent spontaneous abortion (RSA), we used polymerase chain reaction (PCR) amplification, and genotyped 85 women in the case group (women who have had two or more unexplained RSA) and 85 women in the control group (women who have had at least one normal pregnancy). Our results showed that the frequencies of the−14 bp/−14 bp and +14 bp/+14 bp genotypes were reduced in women with RSA, while that of the +14 bp/−14 bp genotype was significantly increased in RSA compared with the control group of normal fertile women; no  significant differences in the  allele frequencies of  the  HLA-G  14-bp polymorphism were observed. These results suggest a possible significance of the HLA-G 14-bp INDEL polymorphism in the outcome of pregnancy. However, further studies on other polymorphic sites in the 3 UTR  and  5′  UTR  regions, as well as monitoring  the  serum  HLA-G  concentration  are necessary in order to determine the potential implications of this marker in our population

Novel multi-layer APPPA microcapsules for oral delivery: preparation condition, stability and permeability

491-497Oral therapy utilizing cell microencapsulation has shown promise in the treatment of many diseases. Current obtainable microcapsule membranes, however, show inadequate stability in the gastrointestinal (GI) environment, thus restricting the general application of live cells for oral therapy. To overcome this limitation, we have previously developed a novel multi-layer alginate/poly-L-lysine/pectin/poly-L-lysine/alginate microcapsule (APPPA) with demonstrated improvement on membrane stability over the frequently reported alginate/poly-L-lysine/alginate (APA) microcapsules. In this study, we further examined the effects of preparation conditions on microcapsule formation, and assessed the membrane strength and GI stability. Results showed that increased membrane strength of the APPPA microcapsules was attained by using pectin with low degree of esterification as the mid-layer material, saline as the solvent for the preparation solutions and washing medium, and 0.1 M CaCl2 as the gelling solution for alginate cores. Resistance of this membrane to the simulated GI fluids was also investigated. Permeability of and release profiles from the APPPA microcapsules were found comparable to the APA microcapsules. These findings suggested that the multi-layer APPPA microcapsule formulation may have potential in oral delivery of proteins, live bacterial cells and other biomedical applications

The Evaluation of Efflux Pump Genes norA, norB and norC Related to Fluoroquinolones Resistance in Staphylococcus aureus Strains Isolated from Blood Infection

Background & Objective: Recently, ciprofloxacin resistance in Staphylococcus aureus strains, due to efflux pumps, has become a significant challenge. Therefore, this study was performed to evaluate the frequency of norA, norB, and norC efflux pump genes and their roles in resistance to ciprofloxacin in clinical isolates of S. aureus. Materials & Methods: A total of one hundred clinical blood samples were collected from patient in Qom hospitals and S. aureus isolates were identified by standard microbiological tests. Antimicrobial susceptibility patterns were determined by the disk diffusion method using CLSI guidelines. Subsequently, the presence of norA, norB, and norC efflux pump genes in ciprofloxacin isolates was detected using the PCR method. Results: Among one hundred clinical samples, 36 S. aureus isolates were recovered and the results of antibiotic susceptibility tests showed that twenty of them were resistant to ciprofloxacin. 15 isolates were resistant to norfloxacin and one isolate was resistant to ofloxacin. Moreover, the norA, norB, and norC genes were found in 58%, 30%, and 41% of ciprofloxacin-resistant isolates, respectively. Conclusion: Based on the results of this study, norA, norB, and norC efflux pumps may play a significant role in the development of resistance to ciprofloxacin in clinical isolates of S. aureus. Detecting these genes may prove useful in suggesting an effective treatment model for infections caused by S. aureus

Diverse genetic causes of amenorrhea in an ethnically homogeneous cohort and an evolving approach to diagnosis

International audienceRESEARCH QUESTION: Premature ovarian insufficiency (POI) is characterised by amenorrhea associated with elevated follicle stimulating hormone (FSH) under the age of 40 years and affects 1-3.7% women. Genetic factors explain 20-30% of POI cases, but most causes remain unknown despite genomic advancements. DESIGN: We used whole exome sequencing (WES) in four Iranian families, validated variants via Sanger sequencing, and conducted the Acyl-cLIP assay to measure HHAT enzyme activity. RESULTS: Despite ethnic homogeneity, WES revealed diverse genetic causes, including a novel homozygous nonsense variant in SYCP2L, impacting synaptonemal complex (SC) assembly, in the first family. Interestingly, the second family had two independent causes for amenorrhea - the mother had POI due to a novel homozygous loss-of-function variant in FANCM (required for chromosomal stability) and her daughter had primary amenorrhea due to a novel homozygous GNRHR (required for gonadotropic signalling) frameshift variant. WES analysis also provided cytogenetic insights. WES revealed one individual was in fact 46, XY and had a novel homozygous missense variant of uncertain significance in HHAT, potentially responsible for complete sex reversal although functional assays did not support impaired HHAT activity. In the remaining individual, WES indicated likely mosaic Turners with the majority of X chromosome variants having an allelic balance of ∼85% or ∼15%. Microarray validated the individual had 90% 45,XO. CONCLUSIONS: This study demonstrates the diverse causes of amenorrhea in a small, isolated ethnic cohort highlighting how a genetic cause in one individual may not clarify familial cases. We propose that, in time, genomic sequencing may become a single universal test required for the diagnosis of infertility conditions such as POI