PLT antigen discrepancy pattern among couples with recurrent abortion.

Background: Recurrent abortion refers to a condition of two or more consecutive pregnancies without known etiology affected by miscarriage before the completion of the 20th week of gestational age. However, several hypotheses have been proposed, but not much data are available concerning the relationship between human platelet antigens (HPAs) polymorphisms and recurrent abortion. This study was conducted to evaluate the genetic differences between HPA-1, -2, -3, -5, and - 15 in Iranian couples with a history of recurrent abortion. Methods: In this cross-sectional study, a total of 74 couples with at least 2 recurrent abortions without any known specified reasons enrolled in the study. HPA polymorphisms genotyping was performed by single-specific primer PCR. Genotype frequency was calculated using the Hardy-Weinberg equation. Results: A total of 39 couples (52.7%) had HPA genotyping partial mismatches. The most common partial mismatch pairs were found concomitantly on both HPA-15a and HPA-15b in three couples (4%), followed by two (2.7%) on HPA-3a and one (1.3%) in each HPA-2b and HPA-5b. There was a deviation from the Hardy-Weinberg equilibrium in the HPA-2 and -5 systems. Conclusion: The present study declared that partial mismatches of HPA-3 and -15 genotypes were common among Iranian couples due to the history of recurrent abortion and approximately half of the couples carried at least one HPA gene that was absent in their partners. Further studies might be helpful to clarify the association between HPA polymorphisms and recurrent abortion, such as an investigation into the alloantibodies against HPAs.

Unraveling the Impact of Blood Transfusion on Transcription Factors Regulating T Helper 1, 2, 17 and Regulatory T cells.

T helper 1 (TH1) and TH2 lymphocytes are the most important components of the immune system affected by blood transfusion. This study aimed`` to evaluate the effect of blood transfusion on gene expression of transcription factors related to the development of TH1, TH2, TH17 and regulatory T cells (Tregs). In this cross-sectional study, 20 patients diagnosed with abdominal aortic aneurysms requiring surgical repair were studied from January 2018 to August 2020. We utilized real-time PCR to evaluate the expression of transcription factor genes associated with TH1, TH2, TH17, and Treg, namely T-box-expressed-in-T-cells (T-bet), GATA-binding protein 3 (GATA-3), retinoid-related orphan receptor (RORγt), and fork head box protein 3 (Foxp3), respectively. The sampling occurred before anesthesia, 24- and 72 hours post-transfusion, and at the time of discharge. The results showed that the T-bet gene expression, compared to the time before transfusion, was significantly decreased 24 hours after blood transfusion and upon discharge while GATA3 genes exhibited a significant reduction both 24 and 72 hours after the transfusion, as compared to the pre-transfusion levels and the time of patient discharge. The Foxp3 gene demonstrated an increase at all study stages, with a notable surge, particularly 72 hours after red blood cell (RBC) transfusion. Conversely, the expression of RORγt gene, consistently decreased throughout all stages of the study. RBC transfusion in abdominal aortic aneurysm patients altered the balance of transcription gene expression of TH1, TH2, TH17, and Treg cells.

A tight interplay between platelet activation and mitochondrial DNA release promotes platelet storage lesion in platelet concentrates.

BACKGROUND AND OBJECTIVES: Platelet storage lesion (PSL) adversely affects the quality of platelet concentrates (PCs). Platelets are prone to activation during storage. Moreover, elevated free mitochondrial DNA (mtDNA) levels in PCs are associated with a higher risk of adverse transfusion reactions. Therefore, we aimed to evaluate the correlation between platelet activation markers and mtDNA release during PC storage. MATERIALS AND METHODS: Six PCs prepared by the platelet-rich plasma method were assessed for free mtDNA copy number using quantitative real-time PCR and CD62P (P-selectin) expression by flow cytometry on days 0 (PC collection day), 3, 5 and 7 of storage. Lactate dehydrogenase (LDH) activity, pH, platelet count, mean platelet volume (MPV) and platelet distribution width (PDW) were measured as well. The correlation between free mtDNA and other PSL parameters, and the correlation between all parameters, was determined. RESULTS: Significant increases in free mtDNA, MPV and PDW, and a significant decrease in platelet count and pH were observed. CD62P expression and LDH activity elevated significantly, particularly on storage days 5-7 and 0-3, respectively. Moreover, a moderate positive correlation (r = 0.61) was observed between free mtDNA and CD62P expression. The r values between free mtDNA and LDH, pH, platelet count, MPV and PDW were 0.81, -0.72, -0.49, 0.81 and 0.77, respectively. CONCLUSION: The interplay between platelet activation and mtDNA release in promoting PSL in PCs may serve as a promising target for future research on applying additive solutions and evaluating the quality of PCs to improve transfusion and clinical outcomes.

Study on genotyping and coinfection rate of human parvovirus 4 among the HTLV-I/II infected blood donors in Khorasan Razavi, Iran.

Human Parvovirus 4 (PARV4) is an emerging virus infecting individuals with other blood-borne diseases. This study aimed to determine the prevalence of PARV4 in confirmed HTLVI/II positive samples from blood donors, assessing PARV4 viral load (DNA) and genotyping. METHODS: A novel qReal-Time PCR, based on a plasmid construct, was developed to simultaneously detect all three PARV4 genotypes using in-house primers and probes. Positive qPCR samples were subjected to nested PCR amplification and subsequent sequencing. Phylogenetic trees were constructed using the Neighbor-joining (N.J.) method. RESULTS: The coinfection rate of PARV4-DNA in HTLVI/II confirmed infected donors, who were previously deferred, was 14.4 % (13 out of 90), with no observed association with donation status (p = 1.0). Phylogenetic analysis indicated that PARV4-positive samples closely resembled genotype 2 in Iran.qPCR quantification demonstrated significant PARV4 viral loads in positive samples, ranging between 104 and 106 DNA copies/mL of serum. CONCLUSION: This study presents the first evaluation of HTLVI/II and PARV4coinfection rates among blood donors. Notably, elevated PARV4-DNA titers were detected in HTLVI/II-positive donors. Given PARV's resistance to standard plasma refinery inactivation methods and the absence of its targeted inactivation, its potential impact remains a concern.

Resveratrol reduces platelet storage lesion by preventing free mitochondrial DNA (mtDNA) accumulation in platelet concentrates during storage.

Platelet activation and mitochondrial damage are among the crucial events leading to the quality reduction of platelet concentrates (PCs) during preparation and storage, called platelet storage lesion. Platelet activation results in the clearance of transfused platelets. Oxidative stress and platelet activation trigger mitochondrial DNA (mtDNA) release into the extracellular milieu which is associated with adverse transfusion reactions. Therefore, we aimed to investigate the effects of resveratrol, an antioxidant polyphenol, on platelet activation markers and mtDNA release. Ten PCs were divided equally into two bags each, one of them was allocated to the control group (n = 10) and another to the case group (resveratrol-treated, n = 10). Free mtDNA level and CD62P (P-selectin) expression level were measured by absolute quantification Real-Time PCR, and flow cytometry on days 0 (the receiving day), 3, 5, and 7 of storage respectively. Moreover, Lactate dehydrogenase (LDH) enzyme activity, pH, platelet count, mean platelet volume (MPV), and platelet distribution width (PDW) were assessed as well. Treatment of PCs with resveratrol can significantly decrease mtDNA release during storage compared to the control. In addition, platelet activation was significantly mitigated. We also observed significantly lower MPV, PDW, and LDH activity in resveratrol-treated PCs compared to the control group on days 3, 5, and 7. Furthermore, resveratrol maintained the pH of PCs on day 7. Resveratrol diminished free mtDNA and maintained biochemical parameters in PCs, possibly by reducing platelet activation. Therefore, resveratrol might be a possible additive solution for improving the quality of stored PCs.

Polymerase chain reaction test for diagnosis of infectious uveitis.

BACKGROUND: To study the clinical utility of broad-range real-time Polymerase Chain Reaction (PCR) assay in patients suspected for infectious uveitis and to analyze the clinical relevance. METHODS: Medical records of patients with uveitis were assessed in whom PCR analysis of intraocular fluids was performed between January 2018 and February 2021. Intraocular samples were investigated for cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), herpes simplex viruses type 1 and 2 (HSV1,2), human T-lymphotropic virus type 1 (HTLV-1), Toxoplasma gondii and also for bacterial 16 S and fungal 18 S/28S ribosomal DNA (rDNA). RESULTS: Aqueous paracentesis and vitreous sampling was done for 151 (81.2%) and 35 (18.8%) patients, respectively. Most of the patients had panuveitis (61.3%). PCR results were positive in 69 out of 186 patients (37%) according to the following order: CMV (18 cases), VZV (18 cases), fungal 18s/28s rDNA (17 cases), HSV (9 cases), bacterial 16s rDNA (3 cases), HTLV-1 (2 cases), and Toxoplasma gondii (2 cases). PCR positivity rate was 5.8% in patients with undifferentiated panuveitis. EBV was not detected at all. Initial treatment was changed in 38 patients (20%) based on PCR results. The overall sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of PCR test for aqueous samples was 82%, 91%, 96%, and 87%, respectively. No significant adverse effect related to sampling was reported. CONCLUSION: PCR analysis of intraocular fluids in patients with suspected infectious uveitis plays an important role in confirming diagnosis or changing treatment with good predictive value. However, routine PCR test in patients with undifferentiated panuveitis in order to rule out possible underlying infectious etiology had low benefit.

Frequency of Human Platelet Antigens -1 to -5 and -15 in Turkmen Blood Donors.

Background: Due to the presence of platelet antigen polymorphisms, human platelet membrane glycoproteins can be identified as an alloantigen or autoantigen. The aim of this study was to determine the frequencies of human platelet antigens (HPAs)-1 to-5 and-15 in Turkmen blood donors and establish a panel of accredited HPAs negative donors as well as an HPA-typed platelet donor registry. Materials and Methods: HPA-1 to-5 and-15 typing was performed by the polymerase chain reaction-sequence-specific primer techniques on 80 unrelated Turkmen donors who were referred to Aq-Qala Blood Transfusion Center in Golestan Province from September 2018 to October 2019. Results: The frequencies of HPA phenotypes were determined as follows: HPA-1aa: 92.5%, HPA-1ab: 7.5%, HPA-2aa: 77.5%, HPA-2ab: 20.0%, HPA-2bb: 2.5%, HPA-3aa: 75.3%, HPA-3ab: 50%, HPA-3bb: 11.2%, HPA-4aa: 100%, HPA-5aa: 78.5%, HPA-5ab: 21.5%, HPA-15aa: 41.2%, HPA-15ab: 56.2% and HPA-15bb: 17.5%. Conclusion: Determining the genotype of HPAs that play an important role in platelet refractory can improve the management of alloimmunization due to the incompatibility of HPAs between the recipients and donors. Therefore, the registration process for national platelet donors can help patients accelerate and improve the quality of transfused platelets.

Potential involvement of miR-183/96/182 cluster-gene target interactions in transdifferentiation of human retinal pigment epithelial cells into retinal neurons.

miR-183/96/182 cluster plays a critical role in the developing retina by regulating many target genes involved in signaling pathways. This study aimed to survey the miR-183/96/182 cluster-target interactions that, potentially contribute to human retinal pigmented epithelial (hRPE) cell differentiation into photoreceptors. Target genes of the miR-183/96/182 cluster were obtained from miRNA-target databases and applied to construct miRNA-target networks. Gene ontology and KEGG pathway analysis was performed. miR-183/96/182 cluster sequence was cloned into an eGFP-intron splicing cassette in an AAV2 vector and overexpressed in hRPE cells. The expression level of target genes including HES1, PAX6, SOX2, CCNJ, and RORΒ was evaluated using qPCR. Our results showed that miR-183, miR-96, and miR-182 share 136 target genes that are involved in cell proliferation pathways such as PI3K/AKT and MAPK pathway. qPCR data indicated a 22-, 7-, and 4-fold overexpression of miR-183, miR-96, and miR-182, respectively, in infected hRPE cells. Consequently, the downregulation of several key targets such as PAX6, CCND2, CDK5R1, and CCNJ and upregulation of a few retina-specific neural markers such as Rhodopsin, red opsin, and CRX was detected. Our findings suggest that the miR-183/96/182 cluster may induce hRPE transdifferentiation by targeting key genes that involve in the cell cycle and proliferation pathways.

The evaluation of reactive oxygen species generation and free mitochondrial DNA in platelet concentrates during 5 days of storage.

Oxidative stress and mitochondrial damage are causes of platelet storage lesions (PSLs). Mitochondrial damage causes mitochondrial DNA (mtDNA) to be released into the extracellular space. MtDNA in platelet concentrates is considered damage-associated molecular patterns (DAMPs) and is one of the major causes of PSLs. The mechanism of mtDNA release in platelet concentrates has not been thoroughly investigated. This study aimed to determine the effect of reactive oxygen species (ROS) on mtDNA release in platelet concentrates during storage. Ten platelet concentrates from healthy donors were obtained in this investigation. Platelet concentrates were prepared by platelet-rich plasma (PRP) and stored at 22 ±â€Š2 C° with gentle agitation. Platelet concentrates were subjected to flow cytometry and real-time PCR to evaluate total ROS and free mtDNA on days 0, 3, and 5 of platelet concentrate storage. Total ROS detected significantly increased from day 0 to day 5 of platelet concentrate storage (P = 0.0079). The mean of copy numbers of free mtDNA on day 0 increased from 3.43 × 106 ±â€Š1.57 × 106 to 2.85 × 107 ±â€Š1.51 × 107 (molecules/µl) on the fifth day of platelet concentrate storage, and it was statistically significant (P = 0.0039). In addition, LDH enzyme activity significantly increased during platelet concentrate storage (P < 0.0001). Also, releasing mtDNA in platelet concentrates was directly correlated with total ROS generation (P = 0.021, r = 0.61) and LDH activity (P = 0.04, r = 0.44). The evidence from this study confirmed the increasing level mtDNA copy numbers in platelet concentrates during storage, and the amount of free mtDNA is directly correlated with ROS generation and platelet lysis during 5 days of platelet concentrate storage. Finally, these changes may be related to DAMPs in the platelet concentrates.

L-carnitine modulates free mitochondrial DNA DAMPs and platelet storage lesions during storage of platelet concentrates.

Platelet storage lesions may occur in Platelet concentrates (PCs) storage time, reducing PCs' quality. Mitochondrial damage causes mitochondrial DNA (mtDNA) to be released into the extracellular space. In this study, we evaluated the effect of L-carnitine (LC) as an antioxidant on free mtDNA DAMPs release in PCs during storage. Ten PCs prepared by the PRP method were studied. The copy numbers of free mtDNA, total reactive oxygen species (ROS), lactate dehydrogenase (LDH) enzyme activity, pH, and platelet counts were measured on days 0, 3, 5, and 7 of PCs storage in LC-treated and untreated platelets. LDH activity was significantly lower than the control group during 7 days of PCs storage (p = 0.041). Also, ROS production decreased in LC-treated PCs compared to the control group during storage (p = 0.026), and the difference mean of ROS between the two groups was significant on day 3, 5, and 7 (Pday3 = 0.02, Pday5 = 0.0001, Pday7 = 0.031). Moreover, LC decreased the copy numbers of free mtDNA during 7 days of storage (p = 0.021), and the difference mean of the copy numbers of free mtDNA in LC-treated PCs compared to the control group was significant on day 5 and 7 (Pday5 = 0.041، Pday7 = 0.022). It seems that LC can maintain the metabolism and antioxidant capacity of PCs and thus can reduce mitochondrial damage and mtDNA release; consequently, it can decrease DAMPs in PCs. Therefore, it may be possible to use this substance as a platelet additive solution in the future.

Comparison mitochondrial DNA DAMPs changes in platelet prepared by apheresis and PRP.

INTRODUCTION: Mt DNA (DAMPs) plays a key modulatory role in immune cells and may also mediate a variety of adverse transfusion reactions. MATERIAL AND METHOD: The cross-sectional study was performed on 22 (PRP) and 14 (SDP) between February 2019 and 2020. mtDNA DAMPs by quantitative real-time PCR was assessed on days 1, 3, and 5 of platelets storage. The data was entered in REST 2009 software, and the amount of fold change was calculated. Multiple t tests were also used. RESULTS: The mtDNA DAMPs fold change in SDP on days 1-3, 1.3 times, on days 3-5, 1.5 times, and on days 1-5, 2.1 times increased. The fold change on days 1-3, 0.8 times, on days 3-5, 0.6 times, and on days 1-5, 0.49 times decreased in PRP products. CONCLUSION: The method of preparation and processing can affect mtDNA DAMPs fold changes.

Validating the HPA-1 to -5 and -15 Detection by Homemade PCR-SSP, Real-Time PCR, and PCR-RFLP Methods.

OBJECTIVE: Human platelet antigens (HPAs) are antigenic determinants on platelet membrane glycoproteins that stimulate the host's immune system and cause platelet destruction. In this study, we share our experience with implementing sequence-specific primer-polymerase chain reaction (PCR-SSP), real-time PCR, and PCR-RFLP (restriction fragment length polymorphism) and the validation process used to evaluate the results. METHODS: At the Ardabil Blood Transfusion Center, 10 samples were obtained from blood donors. Validation using PCR-SSP, real-time PCR, and PCR-RFLP methods for genotyping HPAs was done by sequencing. A commercial DNA sample and a commercial kit were also used for validation. RESULTS: The results of PCR-SSP, TaqMan Real-Time PCR, melting curve analysis (HPA-15), and PCR-RFLP (HPA-3) were 100% consistent with sequencing (gold standard) and commercial kit results. CONCLUSIONS: There was a 100% correlation between repeating the methods and the expected results for repeatability, and no false positives and negatives were observed.

Investigation of human parvovirus B19 prevalence in a large healthy umbilical cord blood donors.

Background and Objectives: Umbilical cord blood (UCB) was used to source hematopoietic stem cells in the past. Despite the apparent advantages of UCB transplantation, virus reactivation poses a considerable danger in allogeneic hematopoietic stem cell transplantation (HSCT). Human Parvovirus B19 is regarded as a potential threat to UCB contamination. This study aimed to evaluate the prevalence of parvovirus B19 in cord blood donors by Semi-Nested PCR. This study is the first largescale report of the B19 DNA in cord blood donors in Iran. Materials and Methods: A total of 691 umbilical cord blood were collected under standard procedure. Then, DNA from buffy coat and plasma were extracted, and semi-nested PCR was performed for all samples. Results: Two out of 691 samples (0.29%) indicated viremia in plasma and buffy coat. Conclusion: In this line, designing and validating a quantitative PCR assay for detection, quantification, and discrimination of Human B19 DNA genotypes of cord blood donors is necessary to enhance the safety of this source of stem cells.

The effect of platelet apheresis collection on some immunological factors in donors using two different apheresis devices.

INTRODUCTION: The aim of this study was to evaluate and compare two different apheresis and changes in some immunological factors in donors. MATERIAL AND METHODS: The cross-sectional study was performed from January 2017 to September 2018. Fifty six male blood donors were randomly divided into two groups. CD4, CD8, and CD25 markers by flow cytometry, and TGFBeta by real-time polymerase chain reaction (RT-PCR) method were done before and 7 days after the apheresis procedure. Independent Sample t-test, Mann-Whitney U Test, Wilcoxon signed ranked test, and Fisher exact test were used. RESULTS: WBC in MCS+ group after donation is significantly higher than before donation (P < 0.05) but no significant difference was seen between MCS+ and Trima groups in these two indicators. But in CD4, CD25, and TGFBeta, there was no significant difference between the two groups. CONCLUSION: There was no significant difference on CD4, CD25, and TGFBeta gene 7 days after donation.

Neural differentiation of human retinal pigment epithelial cells on alginate/gelatin substrate.

Purpose: The development of biomaterials provides potent promise for the regeneration of neuroretinal cells in degenerative eye diseases and retinal tissue engineering. Biomimetic three-dimensional (3D) microenvironments and specific growth factors motivate the differentiation of human retinal pigment epithelial (hRPE) cells toward a retinal neural lineage. In this study, we evaluated alginate/gelatin (A/G) as a substrate for the culture of hRPE cells. Methods: hRPE cells were isolated from neonatal human cadaver globes and cultivated on A/G substrate under different culture conditions, including 30% human amniotic fluid (HAF), 10% fetal bovine serum (FBS), and serum-free Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM/F12). The proliferation of cells in different culture conditions was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and a cell proliferation assay. Immunocytochemistry and real-time PCR were performed to evaluate the effect of the substrate on hRPE cell differentiation. Results: A significant increase in the cell proliferation rate was observed in hRPE cells cultivated on an A/G substrate. Continuous observations demonstrated that hRPE cells formed densely packed, suspended spheroids in DMEM/F12 culture conditions, with dominant transdifferentiation into amacrine cells. Small adherent clusters of hRPE cells in HAF- and FBS-treated cultures represented dedifferentiation toward retinal progenitor cells. These cultures generated amacrine, rod photoreceptors, and bipolar cells. Conclusions: These findings indicated that A/G substrate induced neural retinal cell propagation in cultures and would therefore be promising for RPE-based tissue engineering studies.

Genotyping of Human Platelet Antigen-1 to -5 and -15 by Polymerase Chain Reaction with Sequence-specific Primers (PCR-SSP) and Real-time PCR in Azeri Blood Donors.

Human platelet antigens (HPAs) are glycoproteins on the platelet surface that a single nucleotide mutation in the coding region gene could lead to the variation of different HPA polymorphisms. These antigens have shown variation among different races and may trigger immune responses during blood transfusion and pregnancy. Genotyping of HPAs is useful for managing these reactions and establishing a platelet registry to decrease platelet transfusion reactions. This study aimed to compare allelic and genotype frequencies of human platelet antigens in the Azeri ethnicity by TaqMan Real-time and polymerase chain reaction with sequence-specific primers (PCR-SSP) methods. DNA was extracted from the whole blood of 100 Azeri blood donors in the Ardabil Blood Transfusion Center. Genotyping of HPA-1 to -5 and -15 was performed by TaqMan Real-time PCR, and PCR-SSP and consistency of results were evaluated. The results of PCR-SSP and TaqMan Real-time PCR showed complete consistency. The allele frequencies were 91.5% and 8.5% for HPA-1a and -1b; 88% and 12% for HPA-2a and -2b; 58% and 42 % for HPA-3a and -3b; 100% for HPA-4a; 91% and 9% for HPA-5a and -5b; 56.5% and 43.5% for HPA-15a and -15b alleles. Not incompatibility was detected in HPAs genotyping by PCR-SSP and TaqMan Real-time PCR so that real-time PCR can be used as a robust and quick method for HPA genotyping. We found differences between Azeri blood donors and previously reported HPAs alleles' frequency in other ethnicities in the country. This fact highlights the need for a platelet registry to recruit platelet donors from different ethnicities and increase the number of donors by using faster methods.

Optogenetic control of neural differentiation in Opto-mGluR6 engineered retinal pigment epithelial cell line and mesenchymal stem cells.

In retinal degenerative disorders, when neural retinal cells are damaged, cell transplantation is one of the most promising therapeutic approaches. Optogenetic technology plays an essential role in the neural differentiation of stem cells via membrane depolarization. This study explored the efficacy of blue light stimulation in neuroretinal differentiation of Opto-mGluR6-engineered mouse retinal pigment epithelium (mRPE) and bone marrow mesenchymal stem cells (BMSCs). mRPE and BMSCs were selected for optogenetic study due to their capability to differentiate into retinal-specific neurons. BMSCs were isolated and phenotypically characterized by the expression of mesenchymal stem cell-specific markers, CD44 (99%) and CD105 (98.8%). mRPE culture identity was confirmed by expression of RPE-specific marker, RPE65, and epithelial cell marker, ZO-1. mRPE cells and BMSCs were transduced with AAV-MCS-IRES-EGFP-Opto-mGluR6 viral vector and stimulated for 5 days with blue light (470 nm). RNA and protein expression of Opto-mGluR6 were verified. Optogenetic stimulation-induced elevated intracellular Ca2+ levels in mRPE- and BMS-treated cells. Significant increase in cell growth rate and G1/S phase transition were detected in mRPE- and BMSCs-treated cultures. Pou4f1, Dlx2, Eomes, Barlh2, Neurod2, Neurod6, Rorb, Rxrg, Nr2f2, Ascl1, Hes5, and Sox8 were overexpressed in treated BMSCs and Barlh2, Rorb, and Sox8 were overexpressed in treated mRPE cells. Expression of Rho, Thy1, OPN1MW, Recoverin, and CRABP, as retinal-specific neuron markers, in mRPE and BMS cell cultures were demonstrated. Differentiation of ganglion, amacrine, photoreceptor cells, and bipolar and Muller precursors were determined in BMSCs-treated culture and were compared with mRPE. mRPE cells represented more abundant terminal Muller glial differentiation compared with BMSCs. Our results also demonstrated that optical stimulation increased the intracellular Ca2+ level and proliferation and differentiation of Opto-mGluR6-engineered BMSCs. It seems that optogenetic stimulation of mRPE- and BMSCs-engineered cells would be a potential therapeutic approach for retinal degenerative disorders.

Real Time Polymerase Chain Reaction for Hepatitis B Screening in Donor Corneas in the Central Eye Bank of Iran.

PURPOSE: The aim of this study was to report the results of the use of real-time polymerase chain reaction (PCR) for the diagnosis of hepatitis B virus (HBV) infection in cornea donors at the Central Eye Bank of Iran. METHODS: Between 2014 and 2016, all cornea donors that had negative screening serologic results for hepatitis B (HB) surface antigen, HB surface antibody (Ab), hepatitis C virus Ab, human immune deficiency virus Ab, human T-cell leukemia virus Ab, and syphilis, and positive serology for HB core Ab were subjected to real-time PCR with a detection limit of 400 IU/mL to identify HBV DNA. Positive results for HBV DNA were considered occult HBV infections in these donors. RESULTS: Over the 3-year period, 122 out of 10448 cornea donors had negative screening serologic tests outside of HB core Ab. Of which, 90 cases were subjected to real-time PCR. Occult HBV was detected in 11 cases (12.2%), resulting in the rejection of the corresponding corneas. The remaining 79 cases (87.8%) had negative results for HBV DNA and the corresponding corneas were used for transplantation. CONCLUSION: Implementation of PCR for the detection of occult HBV in cornea donors is necessary to not only increase the security level of cornea donation but also minimize the rejection rate of donors that have isolated HB core Ab reactivity.

The effect of hypoxia preconditioning on the neural and stemness genes expression profiling in human umbilical cord blood mesenchymal stem cells.

In recent years, human umbilical cord blood-derived mesenchymal stem cell (hUB-MSCs) has been regarded as an alternative source for stem cell therapy. In this study, we evaluated the effect of hypoxia preconditioning (HPC) on the expression of Nt-3, GFAP, Nestin, Oct-4 and Nanog genes and proliferative capacity of hUB-MSCs in comparison with normoxic conditions. HPC+Hypoxia protocol includes cultured hUB-MSCs for 15min at 2.5% O2 and after that reoxygenation for 30min at 21% O2 (HPC), and then hypoxia preconditioned hUB-MSCs subjected to 2.5% O2 for 72h (Hypoxia). Conclusively, the results showed that hypoxic preconditioning is an effective strategy for enhancing proliferation capacity of hUB-MSCs, and also can trigger expression of some of the neural genes. In addition, the concept of involvement of oxygen tension in the expression of some of the neural genes of hUB-MSCs would be a good sign of enhanced neural differentiation potential in vitro.

Quantification of viral genome in cord blood donors by real time PCR to investigate human herpesvirus type 8 active infection.

Umbilical cord blood (UCB) is one of the most important sources of hematopoietic stem cells which can be used for transplantation. The transplanted CB stem cells might cause infections in recipients. The aim of this study is to evaluate Human Herpes Virus8 (HHV8) as a Rhadinovirus among CB samples in order to assess safety of cord blood stem cells transplantation. To assess this aim, we surveyed 800 cord blood specimens by Real Time PCR.The overall HHV8 incidence in cord blood mononuclear cells was 1.38% and none of them was in lytic phase of HHV8. The authors suggest further HHV8 study on CB samples for transplantation.