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Background: Platelet-leukocyte aggregates have been implicated in various infectious and inflammatory diseases. The Interferon-induced transmembrane protein 3 (IFITM3) protein plays a role in eliminating viral infections, but its role in the severity of COVID-19 is not well understood. Objectives: We aimed to investigate the correlation between IFITM3 mRNA expression and platelet-monocyte complex levels with the severity of COVID-19, as well as various inflammatory and coagulation markers. Methods: We conducted a cross-sectional study on 54 COVID-19 patients, classified into severe and mild/moderate subgroups. Demographics and laboratory findings were extracted from patients' medical records. We measured IFITM3 mRNA expression in patients' buffy coats using q-RT-PCR and used flow cytometry with CD61 and CD14 markers to measure platelet-monocyte aggregates. Results: No significant difference was found in IFITM3 mRNA expression levels or platelet-monocyte complexes between severe and mild/moderate groups (P = 0.067 and P = 0.056). Lymphocyte counts were significantly higher in the mild/moderate subgroup (21.7 ± 8.9 vs 16.3 ± 10.9, P = 0.02), while neutrophil counts were significantly higher in severe patients (78.3 ± 12.2 vs 72.3 ± 9.9, P = 0.01). Additionally, levels of CRP and LDH were significantly higher in severe COVID-19 patients (P = 0.01 and P = 0.001, respectively). A strong positive correlation was observed between the hospitalization period and CRP, CRP with neutrophils and LDH, as well as between O2 saturation and lymphocytes (P < 0.001, P = 0.0003, P = 0.002, and P = 0.005, respectively). Conclusions: Our findings suggest that IFITM3 gene expression and platelet-monocyte aggregate levels do not correlate with disease outcomes in COVID-19. However, further investigations with larger sample sizes are needed to better understand the mechanisms involved. Monitoring inflammatory and coagulation markers remains important for managing COVID-19 patients. [ABSTRACT FROM AUTHOR]

Myeloid malignancies arise in bone marrow microenvironments and shape these microenvironments in favor of malignant development. Immune suppression is one of the most important stages in myeloid leukemia progression. Leukemic clone expansion and immune dysregulation occur simultaneously in bone marrow microenvironments. Complex interactions emerge between normal immune system elements and leukemic clones in the bone marrow. In recent years, researchers have identified several of these pathological interactions. For instance, recent works shows that the secretion of inflammatory cytokines such as tumor necrosis factor‐α (TNF‐α), from bone marrow stromal cells contributes to immune dysregulation and the selective proliferation of JAK2V617F+ clones in myeloproliferative neoplasms. Moreover, inflammasome activation and sterile inflammation result in inflamed microenvironments and the development of myelodysplastic syndromes. Additional immune dysregulations, such as exhaustion of T and NK cells, an increase in regulatory T cells, and impairments in antigen presentation are common findings in myeloid malignancies. In this review, we discuss the role of altered bone marrow microenvironments in the induction of immune dysregulations that accompany myeloid malignancies. We also consider both current and novel therapeutic strategies to restore normal immune system function in the context of myeloid malignancies. [ABSTRACT FROM AUTHOR]

Background: Hematological abnormalities in COVID-19 infection included quantitative and qualitative changes and should be further characterized. Evaluation for myelodysplastic syndromes (MDS) is usually prompted by abnormal hematologic findings and the presence of dysplastic morphologies. Viral infections are considered to be the cause of dysplastic morphologies and should be considered by morphologists. There are few reports of dysplastic abnormal morphologies in patients with COVID-19 infection. However, such correlations still have to be clarified. Materials and Methods: In the present study, we examined the granulocyte lineage morphological abnormalities in symptomatic RT-PCR-confirmed COVID patients. Peripheral blood samples were collected from 82 patients with symptomatic COVID-19. Blood smears were prepared according to the standard Wright-Giemsa staining procedure. The morphological examination was carried out by two laboratory experts. Results: Blood smear examination revealed common myelodysplastic syndrome (MDS) type abnormalities including but not limited to pseudo-pelger nuclear lobulation (4.8%), hypogranulation (7.3%), Howell-Jolly-like bodies or detached nuclear segments (6.0%) and elongated and thin nuclear filaments (6.0%). One case of abnormal immature granulocyte and ring form nucleus is also evident. Conclusion: Our results accounted for the possibility of active COVID-19 infection in all subjects with granulocyte dysplasia. These results are of practical importance for patients suspected of having myelodysplastic syndromes or disease processes associated with myeloid malignancies. [ABSTRACT FROM AUTHOR]

Background: Polycystic ovary syndrome (PCOS) is a common endocrinological disorder associated with abdominal obesity (AO) and some reproductive complications including low pregnancy rate. Embryo-endometrium cross-talk has a key role in successful embryo implantation and subsequent normal pregnancy rate. The primary objective of this study is to evaluate the decidualization potential of endometrial stromal cells (ESCs) using the embryo condition media (ECM) collected from PCOS patients with AO, compared to ECM of those patients without AO. Materials and Methods: In this experimental study, we measured the capacity of ECM collected from PCOS patients with or without AO for decidualization induction in healthy ESCs after coculture. A total number of 53 embryos from 40 couples belonging to PCOS with AO, PCOS without AO, nonPCOS with AO, and nonPCOS without AO patients, were included in our study. The embryosof four groups were single-cultured up to the blastocyst stage. Their ECM (45λ/well) were pooled and added to healthy ESCs monolayer culture media to investigate their effects on decidualization potential via gene (PRL, IGFBP1, IL1-β, HOXA10, IL-6 and TNF-α) and protein (PRL, IGFBP1, IL1-β) expression analysis and ESCs migration assay. Results: The morphological analysis, migration assay (P≤0.0321), protein (P≤0.0139) and gene expression analysis showed PCOS with AO accounted for the highest gene (PRL, IGFBP1, IL1-β, HOXA10, IL-6, TNF-α) and protein markers (PRL, IGFBP1, IL1-β) (P≤0.05). NonPCOS individuals without AO had the lowest level of both gene and protein decidualization markers (P≤0.05). Conclusion: Considering decidualization as an inflammatory process, a higher level of decidualization markers was associated with a higher inflammatory status created by AO and PCOS, separately. Inflammation may disrupt the process of inflammatory to anti-inflammatory phase required for prevention of pregnancy loss, this could explain the high rate of abortion in these cases. [ABSTRACT FROM AUTHOR]

Hydroxyurea (HU) is an anti-cancer drug that is used for the treatment of hemoglobinopathies as a γ-globin inducer. However, its dose-dependent effects have hampered its clinical reliability. Resveratrol (RSV) is an antioxidant and γ-globin inducer. The present study aimed to assess their combined effects on the γ-globin gene expression and reactive oxygen species (ROS) level of K562 cells. The results indicated that the γ-globin gene expression was approximately two folds higher in the group treated with RSV 50 µM + HU 25 µM in comparison to HU 100 μM alone (***p < 0.001). However, there was an inverse relationship between the expression of γ-globin gene and HU concentration in the combined groups. Furthermore, the combinations of RSV and HU significantly reduced ROS levels compared to single drugs. Overall, the combination of these compounds was an appropriate strategy for increasing γ-globin expression, reducing oxidant levels, and alleviating the adverse effects of HU. [ABSTRACT FROM AUTHOR]

Background: The objective of this study was to describe the association between luteinizing hormone (LH)/follicle-stimulating hormone (FSH) ratio and demographic variables and maturation stage of oocytes in insulin- resistant and insulin-sensitive patients with polycystic ovary syndrome (PCOS) in comparison with control group. Materials and Methods: In this case-control study, 60 patients with in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) indication were subdivided into 3 groups as follow: 20 subjects were assigned to control (fertile women with male infertility history) group, 20 subjects with PCOS were insulin resistant (IR) and 20 subjects with PCOS were insulin sensitive (IS). After puncture, retrieved oocytes were classified into metaphase II (MII) as mature and in metaphase I (MI) or germinal vesicle stage (GV) as immature. Regression analyses were used to explore the association between MII oocyte number and demographic and clinical variables. Results: LH/FSH ratio was significantly higher in PCOS-IR women compared to controls but not significantly different from that of PCOS-IS group. PCOS-IR women had lower MII oocyte number compared with that of controls. According to multiple regression analysis, the number of previous assisted reproductive technology (ART) cycles was negatively associated with the number of MII oocytes. Conclusion: Insulin resistance can be associated with reductions in MII oocyte number in patients with PCOS. [ABSTRACT FROM AUTHOR]

Background: Acute myeloid leukemia (AML) is a heterogeneous clonal disorder characterized by immature myeloid cell proliferation and bone marrow failure. Various genetic and epigenetic factors have been found to be influential in such patients. Methylation silencing of APAF-1, a putative tumor suppressor gene (TSG), has been found in several human malignancies. In this study, we explored the association of APAF-1 methylation status with AML patients. Materials and Methods: We studied the methylation status of APAF-1 gene in 101 AML patients and 50 healthy subjects as controls. Genomic DNA was extracted from leukocytes in peripheral blood or bone marrow and the methylation status of APAF-1 gene promoter was detectedusing methylation-specific PCR (MSP) method with specific methylated and unmethylated primers. Gene expression was analyzed using real time RT-PCR. Results: The prevalence of methylated (MM) and hemi-methylated (MU) CpG dinucleotides within the APAF-1 gene promoter of AML patients was 12 (11.9%) and 45 (44.6%), respectively, while no methylation was detected in the control samples (p < 0.001). Our results showed a higher frequency of methylated APAF1 in FLT3-ITD mutated cases (p=0.04). APAF1 mRNA expression was significantly lower in methylated cases compared with normal cases. Conclusion: The present study indicated the increased frequency of hypermethylation of APAF-1 gene promoter in AML patients. APAF-1 aberrant CpG island methylation was associated with transcriptional downregulation in AML patients. Therefore, promoter methylation of APAF-1 gene could be considered as an epigenetic factor that contributes to the development of AML. [ABSTRACT FROM AUTHOR]

Background: Myelodysplastic syndromes (MDSs) include a diverse group of clonal bone marrow disorders characterized by ineffective hematopoiesis and pancytopenia. It was found that down regulation of APAF1, a putative tumor suppressor gene (TSG),leads to resistance to chemotherapy and disease development in some cancers. In this study, we investigated the relation of APAF1 methylation status with its expression and clinicopathological factors in myelodysplastic syndrome (MDS) patients. Materials and Methods: Methylation Sensitive-High Resolution Melting Curve Analysis (MS-HRM) was employed in studying the methylation of CpG islands in the APAF1promoter region in MDS. Gene expression was analyzed by using real time RT-PCR. Results: 42.6% of patient samples were methylated in promoter region of APAF1 analyzed, while methylation of the genewas not seen in controls (P<0.05). Methylation of APAF1 was significantly associated with the suppression of its mRNA expression (P=0.00). The methylation status of APAF1in advanced-stage MDS patients (80%) was significantly higher than that of the early-stage MDS patients (28.2%) (P=0.001). The difference in frequency of hypermethylated APAF1 gene was significant between good (37.5%) and poor (85.71%) cytogenetic risk groups (P=0.043). In addition, a higher frequency of APAF1 hypermethylation was observed in higher-risk MDS group (69.2%) compared to lower-risk MDS group (34.14%) (P=0.026). Conclusion: Our study indicated that APAF1hypermethylation in MDS was associated to high-risk disease classified according to the IPSS, WHO and cytogenetic risk. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to evaluate the levels of two oxidative stress (OS) markers including lipid peroxide (LPO) and total antioxidant capacity (TAC) in both serum and follicular fluid (FF) of women with endometriosis after puncture. Materials and Methods: In this cross-sectional study, a total number of sixty-three women younger than 40 years old with laparoscopy (gold standard for endometriosis diagnosis) indication underwent in vitro fertilization (IVF) program in the Royan Institute, Tehran, Iran from September 2013 to October 2014. About forty-three patients were diagnosed with endometriosis after laparoscopy. Blood and FF from the leading follicle in each stimulated ovary were obtained at the time of egg retrieval; samples were centrifuged and frozen until assessment. At the time of sample assessment, serum and FF samples were evaluated for the levels of LPO and TAC on spectrophotometery. Results: We observed that women with endometriosis had significantly higher LPO and lower TAC levels in the serum and FF as compared with the control group (P<0.05). Conclusion: It has observed that FF of women with endometriosis, regardless of disease stage, increases the proliferation power of endometrial cells in vitro, we presume that inflammatory reactions-induced OS in ovary may be responsible for proliferation induction ability in FF obtained from women with endometriosis. [ABSTRACT FROM AUTHOR]

Assessment of embryo quality in order to choose the embryos that most likely result in pregnancy is the critical goal in assisted reproductive technologies (ART). The current trend in human in vitro fertilization/embryo transfer (IVF/ET) protocols is to decrease the rate of multiple pregnancies after multiple embryo transfer with maintaining the pregnancy rate at admissible levels (according to laboratory standards). Assessment of morphological feathers as a reliable non-invasive method that provides valuable information in prediction of IVF/intra cytoplasmic sperm injection (ICSI) outcome has been frequently proposed in recent years. This article describes the current status of morphological embryo evaluation at different pre-implantation stages. [ABSTRACT FROM AUTHOR]

Background: During the last three decades hematopoietic stem cells (HSC) have become a standard protocol for the treatment of many hematologic malignancies and non-malignant disorders. Umbilical cord blood (UCB), as a source of HSCs, has many advantages compared with other sources. One major drawback in using this source in treatment of adult patients is the low HSC dose available. Ex vivo expansion of HSCs is a solution to overcome this limitation. In this study we used TEPA, as a Cu chelator, and human bone marrow (BM) mesenchymal stem cells (MSCs) to investigate expansion rate of UCB-HSCs. Materials and methods: CB-HSCs were isolated using miniMACS magnetic separation system. We cultured the enriched CD34+cells in various conditions: culture condition A, supplemented only with recombinant cytokines; culture condition B, supplemented with BM-MSCs as a cell feeder layer and recombinant cytokines; culture condition C, supplemented with recombinant cytokines and TEPA; culture condition D, supplemented with recombinant cytokines, BM-MSCs as a cell feeder layer and TEPA. In order to evaluate the HSC expansion, we performed cell count, analysis of CD34+ expression by flow cytometry, and colony-forming cell assay on Day 10 after culture. Results: The most fold increase in CD34+ cell, total cell, and total colony numbers was observed in culture condition D (110.11 ± 15.3, 118.5 ± 21, and 172.9 ± 44.7, respectively) compared to other conditions. Conclusion: The results showed that co-culture of HSCs with BM-MSCs in the presence of copper chelating agent (TEPA) could dramatically increase expansion rate of UCB-HSCs. Therefore, this strategy could be useful for HSC expansion. [ABSTRACT FROM AUTHOR]

Background: There is a lack of studies regarding the effects of ultrasound (US) and replication of its exposure on pre-implantation events in mammals. Thus, this study assesses the reproductive performance of mouse oocytes that have been obtained from ovaries irradiated with US waves versus non-irradiated ovaries. Also comparision of their parthenogenesis, ovulation, fertilization, and pre-implantation development rates. Materials and Methods: In this experimental study, we divided extracted ovaries into three experimental groups that received the same dosage, but different replicates of radiation for each group. Results were compared with the control and sham groups. Continuous wave (CW) US, at a spatial average intensity of 355 mW/cm2 and a frequency of 3.28 MHz, was administered for 5 minutes to the ovaries at an interval between pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) injections. Statistical analysis was performed using the ANOVA test and the level of significance was determined to be 0.05. Results: Data collection was based on microscopic visualization. According to the obtained results, metaphase II (MII) oocyte numbers and the percentage of blastocysts significantly reduced in the USexposed groups versus the unexposed groups. Fertilization rate was comparable between groups while parthenogenesis was significantly higher in the US-exposed groups compared to the unexposed groups. Conclusion: Structural damage to cells, intracellular organelles and proteins, as well as changes in signaling pathways induced by US may be reasons for some of the observed adverse effects in groups that have received more US exposure. [ABSTRACT FROM AUTHOR]

Objective: Artificial stimulation of mouse oocyte, in the absence of sperm contribution, can induce its parthenogenic activation of oocyte. Ultrasound is one of the newest methods for artificial activation of mammal oocytes, and its successful utilization in pig oocyte activation has been recently reported. Our objective was to assess the effect of ultrasound on mouse oocyte activation. Materials and Methods: Our groups included control group, 3 experimental groups consisting of 1, 2 and 3 repetitions of ultrasound exposure, and 3 sham groups handled similar to experimental groups but ultrasound system was off during treatments. In experimental groups, adult female NMRI mice at the interval between pregnant mare serum gonadotropin (PMSG) and human corionic gonadotropin (hCG) injections, were exposed to continuous ultrasound with 3.28 MHz frequency and peak intensity (Ipk) = mW/cm2. Sixteen hours after injection of hCG, the mice were euthanized and their oocytes were collected; thereafter, parthenogenic oocytes were counted. Results: Data analysis using the ANOVA test shows a significant increase in the number of parthenogenic oocytes in mice with 3 overall exposures to ovarian ultrasound (p<0.05). A significant decrease in the number of metaphase II (MII) oocytes numbers was also seen in mice treated with ultrasound (p<0.05). Conclusion: Ultrasound is thought to induce pores generation in oocyte membranes and provides an easier inward transport of Ca++ into oocytes. This phenomenon can induce meiosis resumption in immature oocytes. With increased exposure repetitions from I to 3 times and greater Ca++ arrival, oocytes can be parthenogenetically activated. [ABSTRACT FROM AUTHOR]

Objective: The extracellular matrix (ECM) of the cumulus oocyte complex (COC) is composed of several molecules that have different roles during follicle development. This study aims to explore gene expression profiles for ECM and cell adhesion molecules in the cumulus cells of polycystic ovary syndrome (PCOS) patients based on their insulin sensitivity following controlled ovarian stimulation (COS). Materials and Methods: In this prospective case-control study enrolled 23 women less than 36 years of age who participated in an intracytoplasmic sperm injection (ICSI) program. Patients were subdivided into 3 groups: control (n=8, fertile women with male infertility history), insulin resistant (IR) PCOS (n=7), and insulin sensitive (IS) PCOS (n=8). We compared 84 ECM component and adhesion molecule gene expressions by quantitative real-time polymerase chain reaction array (qPCR-array) among the groups. Results: We noted that 21 of the 84 studied genes differentially expressed among the groups, from which 18 of these genes downregulated. Overall, comparison of PCOS cases with controls showed downregulation of extracellular matrix protein 1 (ECM1); catenin (cadherin-associated protein), alpha 1 (CTNNA1); integrin, alpha 5 (ITGA5); laminin, alpha 3 (LAMA3); laminin, beta 1 (LAMB1); fibronectin 1 (FN1); and integrin, alpha 7 (ITGA7). In the IS group, there was upregulation of ADAM metallopeptidase with thrombospondin type 1 motif, 8 (ADAMTS8) and neural cell adhesion molecule 1 (NCAM1) compared with the controls (P<0.05). Conclusion: Downregulation of ECM and cell adhesion molecules seem to be related to PCOS. Gene expression profile alterations in cumulus cells from both the IS and IR groups of PCOS patients seems to be involved in the composition and regulation of ECM during the ovulation process. This study highlights the association of ECM gene alteration as a viewpoint for additional understanding of the etiology of PCOS. [ABSTRACT FROM AUTHOR]

Background: Polycystic ovary syndrome (PCOS) is a common endocrinological disorder associated with abdominal obesity (AO) and some reproductive complications including low pregnancy rate. Embryo-endometrium cross-talk has a key role in successful embryo implantation and subsequent normal pregnancy rate. The primary objective of this study is to evaluate the decidualization potential of endometrial stromal cells (ESCs) using the embryo condition media (ECM) collected from PCOS patients with AO, compared to ECM of those patients without AO., Materials and Methods: In this experimental study, we measured the capacity of ECM collected from PCOS patients with or without AO for decidualization induction in healthy ESCs after coculture. A total number of 53 embryos from 40 couples belonging to PCOS with AO, PCOS without AO, nonPCOS with AO, and nonPCOS without AO patients, were included in our study. The embryosof four groups were single-cultured up to the blastocyst stage. Their ECM (45λ/well) were pooled and added to healthy ESCs monolayer culture media to investigate their effects on decidualization potential via gene ( PRL, IGFBP1, IL1-β, HOXA10, IL-6 and TNF-α ) and protein (PRL, IGFBP1, IL1-β) expression analysis and ESCs migration assay., Results: The morphological analysis, migration assay (P≤0.0321), protein (P≤0.0139) and gene expression analysis showed PCOS with AO accounted for the highest gene (PRL, IGFBP1, IL1-β, HOXA10, IL-6, TNF-α) and protein markers (PRL, IGFBP1, IL1-β) (P≤0.05). NonPCOS individuals without AO had the lowest level of both gene and protein decidualization markers (P≤0.05)., Conclusion: Considering decidualization as an inflammatory process, a higher level of decidualization markers was associated with a higher inflammatory status created by AO and PCOS, separately. Inflammation may disrupt the process of inflammatory to anti-inflammatory phase required for prevention of pregnancy loss, this could explain the high rate of abortion in these cases.

Objective: The presence of a sex related metabolic difference in glucose utilization and, on the other hand, different developmental kinetic rates in human preimplantation embryos, has been previously observed, however, the correlation between these two events is unknown. Oxidative stress (OS) induced by higher glucose consumption appears to be a possible cause for the delayed development rate in female embryos. We examined the correlation between glucose consumption and total antioxidant capacity (TAC) concentration in individual embryo culture media for both male and female embryos., Materials and Methods: In this cross-sectional study, we evaluated high quality embryos from 51 patients that underwent intracytoplasmic sperm injection (ICSI) and preimplantation genetic diagnosis (PGD) at the Royan Institute between December 2014 and September 2017. The embryos were individually cultured in G-2TM medium droplets at days 3-5 or 48 hours post PGD. We analysed the spent culture media following embryo transfer for total antioxidant capacity (TAC) and any remaining glucose concentrations through fluorometric measurement by chemiluminecence system which indirectly was used for measurement of glucose consumed by embryos., Results: The results showed that female embryos consumed more glucose which was associated with decreased TAC concentration in their culture medium compared to male embryos. The mean of glucose concentration consumed by the female embryos (30.7 ± 4.7 pmol/embryo/hour) was significantly higher than that of the male embryos (25.3 ± 3.3 pmol/embryo/hour) (P<0.001). There were significantly lower levels of TAC in the surrounding culture medium of female embryos (22.60 ± 0.19 nmol/μl) compared with male embryos (24.74 ± 0.27 nmol/μl, P<0.01)., Conclusion: This finding highlighted the utilization of sex dependent metabolic diversity between preimplantation embryos for non-invasive sex diagnosis and suggests the TAC concentration as a potential noninvasive biomarker for prediction of sex., Competing Interests: There is no conflict of interest in this study., (Copyright© by Royan Institute. All rights reserved.)

Myelodysplastic syndromes (MDSs) are a clonal bone marrow (BM) disease characterized by ineffective hematopoiesis, dysplastic maturation and progression to acute myeloid leukemia (AML). Methylation silencing of HRK has been found in several human malignancies. In this study, we explored the association of HRK methylation status with its expression, clinical parameters and MDS subtypes in MDS patients. To study the methylation status of HRK gene, we applied Methylation Sensitive-High Resolution Melting Curve Analysis (MS-HRM) in MDS patients, as well as healthy controls and EpiTect®PCR Control DNA. Real time RT-PCR was used for gene expression analysis. Methylation frequency in promoter region of HRK in patient samples was 20.37%. Methylation of HRK was significantly related to transcriptional downregulation (P=0.023). The difference in frequency of hypermethylated HRK gene was significant between good (10%) and poor (71.42%) cytogenetic risk groups (P= 0.001), advanced stage MDS patients (66.66%) in comparison with early stage MDS patients (2.56%) (P= 0.00), higher- risk MDS group (61.53%) and lower- risk MDS group (7.31%) (P= 0.00). HRK hypermethylation was associated with advanced- stage MDS and downregulation of HRK gene may play a role in the progression of MDS.

Objective: The abdominal form of obesity is prevalent in women with polycystic ovary syndrome (PCOS). Visceral fat accumulation seems to play an important role in etiology of PCOS. In this cross-sectional study we evaluated the association of oxidative stress (OS) induced with PCOS and abdominal obesity in serum and follicular fluid (FF) of infertile women., Study Design: A total of 80 women younger than 37 years old undergoing an IVF program were studied in the same period of time from September 2012 to October 2013. Blood serum and FF obtained from 40 women with PCOS (diagnosed by the Rotterdam 2004 criteria) and 40 women without PCOS undergoing IVF were evaluated for two OS markers: lipid peroxide (LPO) and total antioxidant capacity (TAC), after puncture. The patients were divided into 4 groups on the basis of presence of PCOS and waist-to-hip ratio (WHR) or abdominal obesity (OA)., Results: Healthy and PCOS women with abdominal obesity had significantly higher amounts of LPO in the serum and FF as compared with women without abdominal obesity. LPO concentration in FF was significantly lower than in serum and corroborates the hypothesis that the germinal cells have a potent antioxidant mechanism. We also found that LPO concentration in the PCOS group associated with AO had an increasing trend vs. those AO patients without PCOS but this difference was not significant, so the increase in LPO level was approximately independent of PCOS. Based on our results, the association and interaction between PCOS and AO can lead to TAC concentration reduction in patients., Conclusions: Abdominal obesity can induce local and systemic oxidative stress in PCOS and non-PCOS patients. We suggest that PCOS-induced disorders are likely to be exacerbated in the presence of abdominal obesity., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Background: During the last three decades hematopoietic stem cells (HSC) have become a standard protocol for the treatment of many hematologic malignancies and non-malignant disorders. Umbilical cord blood (UCB), as a source of HSCs, has many advantages compared with other sources. One major drawback in using this source in treatment of adult patients is the low HSC dose available. Ex vivo expansion of HSCs is a solution to overcome this limitation. In this study we used TEPA, as a Cu chelator, and human bone marrow (BM) mesenchymal stem cells (MSCs) to investigate expansion rate of UCB-HSCs., Materials and Methods: CB-HSCs were isolated using miniMACS magnetic separation system. We cultured the enriched CD34(+)cells in various conditions: culture condition A, supplemented only with recombinant cytokines; culture condition B, supplemented with BM-MSCs as a cell feeder layer and recombinant cytokines; culture condition C, supplemented with recombinant cytokines and TEPA; culture condition D, supplemented with recombinant cytokines, BM-MSCs as a cell feeder layer and TEPA. In order to evaluate the HSC expansion, we performed cell count, analysis of CD34(+) expression by flow cytometry, and colony-forming cell assay on Day 10 after culture., Results: The most fold increase in CD34(+) cell, total cell, and total colony numbers was observed in culture condition D (110.11 ± 15.3, 118.5 ± 21, and 172.9 ± 44.7, respectively) compared to other conditions., Conclusion: The results showed that co-culture of HSCs with BM-MSCs in the presence of copper chelating agent (TEPA) could dramatically increase expansion rate of UCB-HSCs. Therefore, this strategy could be useful for HSC expansion.