Natural killer cell biology and its effect on graft versus host disease

Natural killer (NK) cells were originally described in terms of their function. NK cells are of lymphoid origin and are found in the peripheral blood, spleen, and bone marrow, as well as other tissues. These cells are large, radio-resistant and granular lymphocytes that represent an important arm of innate immunity and are thought to play a critical role in the immune surveillance against tumors and virally infected cells. Allogeneic bone marrow transplantation (BMT) has proven to be an effective treatment for hematologic malignancies and some solid tumors. One of the major challenges of allo-stem cell transplantation (SCT) is to reduce the incidence and severity of GVHD while boosting the graft-versus-leukemia (GVL) effect. In the setting of allo-SCT, the reconstitution of NK cells is of notable interest due to their known capability to induce GVL without GVHD. Clinical applications of NK cells have been inspired by recognition of their potent anticancer activity. These studies discussed a solid basis for development of future NK cell trials for cancer therapy by minimizing risks and toxicities

microRNA-15b target Sall4 and diminish in vitro UCB-derived HSCs expansion

Hematopoietic Stem Cells (HSCs) are cells that have the ability to self-renewal and differentiate into all of hematopoietic lineages. The lack of donors and unavailable efficient protocols for ex vivo expansion of HSCs, are obstacles in successful cell therapies. MicroRNAs (also refer as miRNAs or miRs) have significant roles in hematopoiesis; they can effect on HSCs expansion, maintaining undifferentiated state, self-renewal and differentiation. Recently attentions have been given to these small regulatory molecules to utilize them in order to expand HSCs. Using bioinformatics analysis we identified Sall4 as putative target of miR-15b and miR-219-5p. Relative expression levels of miRNAs and Sall4 were evaluated by qRT-PCR. Here we show 247-fold and 4.2-fold increasing Sall4 expression level compared to control group in CD34+ cells nucleofected by anti-miR-15b and anti-miR-219-5p, respectively. These data showed that anti-miR-15b can promote clonogenic capacity of HSCs and also we found that miR-15b alone was able to increase the number of CD34+HSCs in vitro by more than 2 fold by targeting Sall4. Moreover, level of CD34 marker in HSCs nucleofected by anti-miR-15b increased more than 50 %. Our analysis showed no statistically difference in mRNA level of Sall4 after nucleofection of anti-miR-219-5p. Sall4 is a factor capable of enhancing HSC expansion significantly. We demonstrated that inhibition of miR-15b can enhance ex vivo expansion of UCB-derived HSCs and also expression of Sall4 allowed expansion and preserve self- renewal of CD34+ HSCs

Genetic study of 45 big hearing loss pedigrees and GJB2 gene mutations frequency in Chaharmahal va Bakhtiari province, Iran, 2008

Background and aim: Hearing loss is the most common sensorineural disorder in human. Despite the contribution of several different genes in causing deafness, mutations in the GJB2 gene have been involved in deafness in many populations. This study aims to investigate genetic epidemiology and frequency of GJB2 gene mutations in 45 big deaf pedigrees. Methods: In this genetic epidemiology study we have investigated 45 big deaf pedigrees concerning inheritance patterns, consanguinity and diversity of deafness severity among siblings using data collected by questionnaires and audiograms. We examined also the frequency and profile of GJB2 gene mutations in 45 probands using direct sequencing strategy. Results: Our study revealed 73% of consanguinity in the deaf pedigrees studied. The most common type of first cousins marriage was found between first cousins who were the children of two brothers. We found autosomal recessive and X-linked recessive pattern in 94-97% and 3-6% of the pedigrees studied respectively. Regarding molecular analysis, GJB2 mutations were found in 11% of population studied including 35delG, 167delT, 299-300delAT and 363delC. Conclusion: A high rate of consanguineous marriage determined in this study could raise the rate of autosomal recessive patterns up to 94-97% of the overall pedigree and would be the main cause to prepare the way for congenital deafness. This study revealed a low contribution of GJB2 gene mutations in causing deafness in Chaharmahal va Bakhtiari province

Gene Expression Analysis of SOX2, NANOG, KLF4, OCT4, and REX1 Genes in Cord Blood Mononuclear Cells Treated with External Stresses

Background: Induced pluripotent stem cells (iPSCS) can be obtained from autologous cells for therapeutic purposes. So, far, many studies have been done to produce induced pluripotent cells by transferring specific pluripotency genes using different methods. In this study, pluripotency gene expression induced by external stresses was assessed in cord blood mononuclear cells.Methods: In this experimental study, mononuclear cells were isolated from umbilical cord blood. Isolated cells were divided into three groups. The first group had been exposed to HCL (pH 5.7) for 25 minutes and then transferred to the medium with normal pH. The second group was triturated with hamilton syringe for 15 min (external pressure), and the last group was considered the control group and did not receive treatment. Then, total RNA was extracted on Day 7. Gene expression of OCT4, SOX2, NANOG, REX1, and KLF4 was evaluated using qRT-PCR.Results: Gene expression of OCT4, NANOG, REX1, and KLF4 was increased after exposure to acidic pH and external pressures in comparison with control cells (P < 0.05). SOX2 gene expression was decreased in cells exposed to acidic pH but increased by external pressure.Conclusions: Exposure of umbilical cord blood mononuclear cells to acidic pH and external pressure lead to re-activation of pluripotency genes in mature cells. These findings indicate that mature cells may be reprogrammed with manipulation of environmental conditions

Gene Expression Analysis of SOX2, NANOG, KLF4, OCT4, and REX1 Genes in Cord Blood Mononuclear Cells Treated with External Stresses

Background: Induced pluripotent stem cells (iPSCS) can be obtained from autologous cells for therapeutic purposes. So, far, many studies have been done to produce induced pluripotent cells by transferring specific pluripotency genes using different methods. In this study, pluripotency gene expression induced by external stresses was assessed in cord blood mononuclear cells.Methods: In this experimental study, mononuclear cells were isolated from umbilical cord blood. Isolated cells were divided into three groups. The first group had been exposed to HCL (pH 5.7) for 25 minutes and then transferred to the medium with normal pH. The second group was triturated with hamilton syringe for 15 min (external pressure), and the last group was considered the control group and did not receive treatment. Then, total RNA was extracted on Day 7. Gene expression of OCT4, SOX2, NANOG, REX1, and KLF4 was evaluated using qRT-PCR.Results: Gene expression of OCT4, NANOG, REX1, and KLF4 was increased after exposure to acidic pH and external pressures in comparison with control cells (P < 0.05). SOX2 gene expression was decreased in cells exposed to acidic pH but increased by external pressure.Conclusions: Exposure of umbilical cord blood mononuclear cells to acidic pH and external pressure lead to re-activation of pluripotency genes in mature cells. These findings indicate that mature cells may be reprogrammed with manipulation of environmental conditions

Evaluation of hematopoietic stem cell expansion in the presence of garcinol

Objective: The application of human cord blood (hCB) is limited to children by using relatively small volume of cord blood that does not contain enough hematopoietic stem cells (HSCs). So, efforts for applying cord blood stem cells in transplantation have led to establishment of some approaches for ex vivo expansion of HSCs such as garcinol. Materials and Methods: CD133+ HSCs were separated by a magnetic-activated cell sorting (MACS) system. Isolated cells were cultured with different doses of garcinol, SCF, TPO and FLT-3L. The optimal dose of garcinol for ex vivo expansion of HSCs was determined by direct counting. Flow cytometry was used to evaluate the expression of CD133 marker to check the ability of garcinol in maintenance of HSCs. Colony forming cell (CFC) assay was performed to evaluate clonogenic capability of treated cells. The level of expression of CXCR4 gene was evaluated by RT-PCR. Data were analyzed using Student’s t test. Results: Our results showed that CD133+ HSCs in the presence of garcinol (5-10 µM) had high expansion activity and cell counting showed that the number of cells in treated group was higher than control group (1.9 –fold) and CFC assay showed that the number of colonies following treatment with garcinol had 1.3-fold increase. Treatment of HSCs with garcinol resulted in 9.6-fold increase in terms of CXCR4 expression in comparison to control group. Conclusion: The present study showed that garcinol can improve ex vivo expansion of HSCs and enhance their potential for homing to bone marrow

DFNB59 Gene Mutation Screening Using PCR-SSCP/HA Technique in Non-syndromic Genetic Hearing Loss in Bushehr Province

Background: Hearing impairment (HI) is the most prevalent Neurosensory disorder which is heterogenous and can also occur due to environmental causes. The majority of hearing deficiencies are of genetic origin affecting about 60% of the HI cases. A novel gene DFNB59 encodes pejvakin has been recently shown to cause deafness. This study aims to determine the frequency of DFNB59 gene mutations in coding region the gene in Bushehr province. Methods: In this descriptive experimental study, we investigated the presence of DFNB59

Relationship among five‐factor personality traits and psychological distress with acoustic analysis

Abstract Objectives The relationship between personality traits and psychological distress with acoustic characteristics was investigated in the present study, regarding the existence of dysphonia, abnormal overall voice quality (AOVQ), and dysphonia type. Methods Fifty‐five participants with dysphonia and 64 participants without dysphonia completed NEO Five‐Factor Inventory and Depression, Anxiety, and Stress Scale‐21. Jitter, shimmer, noise‐to‐harmonic ratio (NHR), cepstral peak prominence (CPP), and cepstral peak prominence‐smoothed (CPPS) were calculated in sustained vowel /a/ by Praat. Three expert speech and language pathologists divided participants with dysphonia into mild, moderate, and severe, based on the AOVQ. Pearson and Spearman correlation tests were performed by IBM SPSS Statistics. Results The findings were indicative of large correlations between agreeableness with CPP, conscientiousness with shimmer, depression with jitter and shimmer, and anxiety with shimmer in patients with functional dysphonia (p  0.05). Conclusion In participants with functional dysphonia, personality traits and psychological distress can provide some information about acoustic characteristics and vice versa. Level of Evidence 3

بررسی ارتباط میان تیپ شخصیتی دانشجویان دندانپزشکی و ترس دندانپزشکی کودکان 7-9 ساله در دانشکده ی دندانپزشکی شهید بهشتی

Objectives Children’s dental fear is a major issue in avoidance of dental visits, which exacerbates dental health problems and is associated with multiple factors. The dentist’s behavior is one of the influential factors in dental fear. This study aimed to evaluate the relationship between dentistry students’ personality type and children’s dental fear. Methods This descriptive, analytical, cross-sectional study was performed on 30 dentistry students and 34 children in the pediatric ward of Shahid Beheshti Dental School, Tehran, Iran. The children’s age was in the range of 7-9 years. To evaluate the students’ personality type, Bortner questionnaire was used, and to investigate children’s dental fear, the Dental Subscale of the Children's Fear Survey Schedule (CFSS-DS) was employed. The children completed the questionnaire twice, once before and once after treatment with the assistance of their parents. Fisher’s exact test (P<0.05), independent t-test (P<0.05), and ANCOVA (P<0.05) tests were performed for statistical analysis. This study was approved by the Ethics Committee of Shahid Beheshti School of Dentistry (ethical ID: IR.SBMU.DRC.REC.1398.176). Results According to the students’ responses to the Bortner questionnaire, they were divided into two groups: 18 students with type A personality and 12 students with type B personality. The children were also divided into two groups. Twenty-two children were treated by dentists with type A personality (group A), and twelve children were treated by dentists with type B personality (group B). Changes in the mean CFSS-DS score following dental treatment were +2.09±7.7 and -2.17±4.95 in group A and group B, respectively. The mean change in the CFSS score of children in group A was higher than that of children in group B, although the difference was not significant (P=0.096). Conclusion The personality type of dentistry students does not significantly affect children’s dental fear.چکیدهسابقه و هدف : ترس دندانپزشکی در کودکان عامل بسیار مهمی در عدم مراجعه به دندانپزشکو در نتیجه مشکلات سلامت دهان و دندان کودکان است که متاثر از چندین فاکتور می باشد . یکیاز این موارد تاثیررفتار دندانپزشک به عنوان یک عامل مستقیم می باشد . بنابراین هدف از اینمطالعه بررسی تاثیر تیپ شخصیتی دانشجوی دندانپزشکی بر روی ترس دندانپزشکی بیماراطفال است .مواد و روش : این مطالعه توصیفی-تحلیلی, مقطعی بر روی 30دانشجوی رشته دندانپزشکیترم 11و 34کودک مراجعه کننده به بخش اطفال دانشکده دندانپزشکی شهید بهشتی انجام شد .محدوده سنی همه ی کودکان 9-7سال بود . جهت سنجش تیپ شخصیتی دانشجویان ازپرسشنامه ی بورتنر و جهت سنجش نمره ی ترس دندانپرشکی کودکان از پرسشنامه یCFSS-DSاستفاده شد . پرسشنامه ی CFSS-DSدر دو نوبت , یک بار قبل از درمان و یکبار بلافاصله بعد از درمان توسط کودک و به کمک والدین وی پر شد . به منظور مقایسه ی بینتفاوت نمره ی پرسشنامه ی CFSS-DSکودکان و نمره ی تیپ شخصیتی دانشجویان و سن وجنس کودکان از مدل سازی رگرسیونی استفاده شد .یافته ها : دانشجویانی که در مطالعه شرکت کردند به دو تیپ شخصیتی 18( Aنفر) و 12( Bنفر) و همچنین 34نفر بیمار که 22نفر توسط تیپ شخصیتی Aو 12نفر توسط تیپ Bدرمانشدند , تقسیم شدند. در این مطالعه تغییرات میانگین نمرات CFSS-DSکودکان در بعد از درمان نسبت به قبل ازدرمان در گروه تیپ شخصیتی +2.09 , Aو در گروه تیپ شخصیتی -2.16 , Bبه دست آمد .نتیجه گیری : این مطالعه نشان داد تیپ شخصیتی دانشجویان دندانپزشکی بر روی ترسدندانپزشکی کودکان تاثیر دارد .واژگان کلیدی : ترس دندانپزشکی کودکان , تیپ شخصیتی , دانشجویان دندانپزشک

Possible involvement of miRNAs in tropism of Parvovirus B19

Human Parvovirus B19 (PVB19) is one of the most important pathogens that targets erythroid lineage. Many factors were mentioned for restriction to erythroid progenitor cells (EPCs). Previous studies showed that in non-permissive cells VP1 and VP2 (structural proteins) mRNAs were detected but could not translate to proteins. A bioinformatics study showed that this inhibition might be due to specific microRNAs (miRNAs) present in non-permissive cells but not in permissive EPCs. To confirm the hypothesis, we evaluated the effect of miRNAs on VP expression. CD34+ HSCs were separated from cord blood. Then, CD34+ cells were treated with differentiation medium to obtain CD36+ EPCs. To evaluate the effect of miRNAs on VP expression in MCF7 and HEK-293 cell lines (non-permissive cells) and CD36+ EPCs, dual luciferase assay was performed in presence of shRNAs against Dicer and Drosha to disrupt miRNA biogenesis. QRT-PCR was performed to check down-regulation of Dicer and Drosha after transfection. All measurements were done in triplicate. Data means were compared using one-way ANOVAs. MicroRNA prediction was done by the online microRNA prediction tools. No significant difference was shown in luciferase activity of CD36+ EPCs after co-transfection with shRNAs, while it was significant in non-permissive cells. Our study revealed that miRNAs may be involved in inhibition of VP expression in non-permissive cells, although further studies are required to demonstrate which miRNAs exactly are involved in regulation of PVB19 replication