Assessment of Standard Operating Procedures (SOPs) Preparing Hygienic Condition in the Blood Donation Centers during the Outbreak of COVID-19.

Background: The coronavirus disease 2019 (COVID-19) outbreak has led to an alteration in hygienic conditions. In this situation, improving standard operating procedures (SOPs) in blood donation centers is critical. The purpose of this study was the assessment of SOPs in the blood donation centers during the outbreak of COVID-19 by regular blood donors as external audits. Materials and Methods: Regular donors were selected as external inspectors in 31 provinces of Iran. The questionnaire containing 10 closed questions was provided to assess the hygienic SOPs of blood transfusion centers in the prevention of COVID-19 transmission. Comparison and evaluation of questionnaires were conducted by assigning an importance coefficient (IC) score to each question. Results: Assessment of SOPs in blood donation departments by regular donors in 31 provinces of Iran showed that 18 centers (58.1%) received IC scores >10(Strong performance), seven centers (22.6%) received the range of IC scores between7-10(acceptable performance), and six centers (19.4%) received IC scores <7(poor performance). The difference in IC scores between provinces was not statistically significant. Conclusion: This study confirms that the assessment of blood donation centers through regular blood donor inspection is a reliable method to identify the strengths and weaknesses of blood transfusion center services and ultimately leads to corrective intervention and improvement of hygienic SOPs to prevent COVID-19 transmission.

The impact of pre-donation viral markers screening of new blood donors on blood safety.

BACKGROUND: The risk of transfusion-transmissible infections (TTIs) remains a concern in transfusion medicine. Since the rate of infection among first-time blood donors is higher than repeated donors, strategies to enhance blood safety can focus on new donors. The aim of the study was to investigate the effect of pre-donation viral screening of new donors on blood safety. METHODS AND MATERIALS: The pre-donation screening of new donors was implemented in the Kurdistan blood center. In this program, new donors who met the blood donation criteria were informed about the program and only a blood sample was donated for HBs Ag, HCV Ab, and HIV Ab testing. New donors with negative results were invited to donate blood after 12 weeks. A unit of blood was collected from eligible returned donors. Laboratory tests were performed again using the same methods. Finally, the prevalence of confirmed positive TTI results among donated blood in Kurdistan blood center was compared before and after the establishment of program. RESULTS: During the study, 4,434 new donors were screened for viral markers. A total of 41 new donors (0.92%, 95% CI, 0.007-0.13) had repeatedly reactive results and infection was confirmed in blood sample of 24 donors (0.54%, 95% CI, 0.003-0.008). Overall, 56% of new donors returned for blood donation. Prevalence of confirmed TTIs markers in collected blood units was 0.27% and 0 before and after implementing program, respectively. CONCLUSIONS: This study indicated that Pre-donation screening can reduce the risk of TTI transmission by identifying infected donors at the pre-donation phase.

Therapeutic effects of mesenchymal stem cells-conditioned medium derived from suspension cultivation or silymarin on liver failure mice.

BACKGROUND: Common treatments of liver disease failed to meet all the needs in this important medical field. It results in an urgent need for proper some new adjuvant therapies. Mesenchymal stem cells (MSCs) and their derivatives are promising tools in this regard. We aimed to compare the Silymarin, as traditional treatment with mesenchymal stem cell conditioned medium (MSC-CM), as a novel strategy, both with therapeutic potentialities in term of liver failure (LF) treatment. METHODS AND RESULTS: Mice models with liver failure were induced with CCl4 and were treated in the groups as follows: normal mice receiving DMEM-LG medium as control, LF-mice receiving DMEM-LG medium as sham, LF-mice receiving Silymarin as LF-SM, and LF-mice receiving MSC sphere CM as LF-MSC-CM. Biochemical, histopathological, molecular and protein level parameters were evaluated using blood and liver samples. Liver enzymes, MicroRNA-122 values as well as necrotic score were significantly lower in the LF-SM and LF-MSC-CM groups compared to sham. LF-SM showed significantly higher level of total antioxidant capacity and malondialdehyde than that of LF-MSC-CM groups. Sph-MSC-CM not only induced more down-regulated expression of fibrinogen-like protein 1 and receptor interacting protein kinases1 but also led to higher expression level of keratinocyte growth factor. LF-MSC-CM showed less mortality rate compared to other groups. CONCLUSIONS: Hepato-protective potentialities of Sph-MSC-CM are comparable to those of Silymarin. More inhibition of necroptosis/ necrosis and inflammation might result in rapid liver repair in case of MSC-CM administration.

Source Plasma Donation: The Experience of the Iranian Blood Transfusion Organization.

Background: A declining need for red blood cells coupled with strengthening demand for plasma-derived medicines has led to a strong focus on moving whole blood donors to plasmapheresis. The purpose of this study was to evaluate the four-year policies of the Iranian Blood Transfusion Organization (IBTO) in terms of plasmapheresis recruitment of first-time donors and its effect on plasmapheresis outcome.   Materials and Methods: Plasmapheresis data related to 16 centers from 2016 to 2019 was obtained from IBTO software. This information includes; (1) blood donation number, (2) plasmapheresis donation number, (3) number of plasmapheresis donors, (4) plasmapheresis donor demographic data, (5) plasmapheresis donor status, (6) frequency of plasma donation for each donor, (7) volume of plasma and (8) the prevalence of transfusion-transmissible infections (TTIs) in plasmapheresis donors. Results: The result of this study demonstrated that plasmapheresis collection centers have recruited 85,515 (91%) first-time and 8,595(9%) regular and repeated donors from 2016 to 2019 years. Plasmapheresis to blood donation index was increased from 0.2% in 2016 to 4.9% in 2019. The mean donation number was 2 times per year. The trend of the yearly Whole Blood Donation (WBD) Index decreased from 26.69 to 24.11/1000 for the general population. The total volume of collected source plasma was 49,203 liters during this period. However, 46,000 liters of recovered plasma were decreased due to less WBD. Furthermore, the results indicated that the prevalence of HCV was significantly higher in first-time donors compared to repeated and regular donors (P = 0.000). Conclusion: It is concluded that during four years, the net volume of plasma did not increase and plasmapheresis led to reducing WBD in our country. Moreover, first-time plasmapheresis donors can be associated with challenges such as increasing screening costs and compromising the safety of plasma resources. Therefore IBTO decided to stop the project and focus on its main role to prepare safe and sufficient blood components through WB collection and also single donor platelet and concurrent plasma by plateletpheresis.

Leukocyte reduction filters as an alternative source of peripheral blood leukocytes for research

ABSTRACT Introduction: Peripheral blood leukocytes are a suitable cell model for science research. However, blood samples from healthy volunteers are limited in volume and difficult to obtain due to the complexity of volunteer recruitment. Objective: Therefore, it is urgent to find an alternative source of peripheral blood leukocytes. Method: One of the possibilities is the use of leukocyte reduction filters (LRFs) in blood banks that is used for preparation of leukoreduced blood products. More than 90% of the leukocytes are trapped in the leukofilters allowing the desired blood product to pass through. Results: It has been reported that the biological function of leukocytes collected from the filters are no different from those isolated from buffy coats, leukapheresis products and whole blood (WB) cells. Moreover, LRFs are waste products that are discarded after leukoreduction. Conclusion: Thus, leukofilters represent an economic source of human cell populations that can be used for a variety of investigative purposes, with no cost. In the present study, we reviewed the different usage of LRFs in the research, clinical and commercial applications.

Investigation of the Osteopontin isoforms expression in patients with acute myeloid leukemia.

Acute myeloid leukemia (AML) is one of the major hematological malignancies. Advances in molecular research have greatly improved our understanding of the process of leukemia formation in AML. Osteopontin (OPN) is a novel molecule that mediates critical processes for cancer progression. The aim of this study was to investigate the relative expression of OPN gene isoforms in AML patients on days 0, 14, and 28 after chemotherapy. The bone marrow samples were collected from 40 newly diagnosed AML patients (24 male and 16 female with a mean age of 30 years) at the initial time of diagnosis, 14 and 28 days after treatment. The peripheral blood samples of 10 healthy individuals were also collected as the control group. The expression of OPN isoforms was investigated by Real-Time Quantitative PCR. The expression of VEGFc/STAT3/CXCR4 was also investigated by Real-Time PCR. Findings indicated that OPNb and OPNc isoforms had significantly overexpression in AML patients on 14 and 28 days after treatment compared to normal samples (P < 0.05). The level of OPNb and OPNc isoforms was increased significantly in M0, M1, and M2 subgroups with overexpression of VEGFc/STAT3/CXCR4, 28 days after starting chemotherapy (P < 0.05). Our results suggested that OPNb and OPNc isoforms play a major role in cancer relapse. Therefore, they can be used as a valuable prognostic and diagnostic biomarker for relapse of the AML disease. However, these findings need confirmation with further studies.

Leukocyte reduction filters as an alternative source of peripheral blood leukocytes for research.

INTRODUCTION: Peripheral blood leukocytes are a suitable cell model for science research. However, blood samples from healthy volunteers are limited in volume and difficult to obtain due to the complexity of volunteer recruitment. OBJECTIVE: Therefore, it is urgent to find an alternative source of peripheral blood leukocytes. METHOD: One of the possibilities is the use of leukocyte reduction filters (LRFs) in blood banks that is used for preparation of leukoreduced blood products. More than 90% of the leukocytes are trapped in the leukofilters allowing the desired blood product to pass through. RESULTS: It has been reported that the biological function of leukocytes collected from the filters are no different from those isolated from buffy coats, leukapheresis products and whole blood (WB) cells. Moreover, LRFs are waste products that are discarded after leukoreduction. CONCLUSION: Thus, leukofilters represent an economic source of human cell populations that can be used for a variety of investigative purposes, with no cost. In the present study, we reviewed the different usage of LRFs in the research, clinical and commercial applications.

Tissue engineering applications in breast cancer.

In Iran, breast cancer (BC) is the most prevalent cancer among women. The standard treatment for this cancer is partial or total removal of breast tissue, followed by chemotherapy and radiation. Tissue engineering (TE) has made new treatments for tissue loss in these patients by creating functional substitutes in the laboratory. In addition, cancer biology combined with TE provides a new strategy for evaluation of anti-BC therapy. Several innovations in TE have led to the design of scaffold or matrix based culture systems that more closely mimic the native extracellular matrix (ECM). Currently, engineered three-dimensional (3D) cultures are being developed for modelling of the tumour microenvironment. These 3D cultures fulfil the need for in vitro approaches that allow an accurate study of the molecular mechanisms and a better analysis of the drugs effect. In the present study, we review recent developments in utilising of TE in BC. Moreover, this review describes achievements of Iranian researchers in the field of breast TE.

Evaluation of anticancer activity of α-defensins purified from neutrophils trapped in leukoreduction filters.

AIMS: The α-defensins or human neutrophil peptides (HNP 1-3) that exist in azurophilic granules are found to have anticancer activity. The pattern of disulfide bonds in α-defensins is crucial for the functional properties. Therefore, synthesis using the chemical and recombinant approaches is a challenging. A safe source for the production of α-defensins can be the use of leukoreduction filters in blood banks that contain large quantities of neutrophils and are discarded after use. The aim of this study was to purify α-defensins from neutrophils trapped in leukofilters and to investigate its anticancer activity. MATERIALS AND METHODS: Immunoprecipitation was performed to purify α-defensins and the presence of protein was confirmed by Western Blot. The Jurkat T-cell line was incubated with different concentrations (5, 10 and 15 µg/ml) of purified HNP1-3 for 16 h. Cell viability was measured using a WST-1 assay and apoptosis was analyzed for Annexin V/PI markers. Caspase-3/7 activity was determined using fluorescence assay. The effects of purified α-defensins were compared to commercial HNP 1-3. KEY FINDINGS: Purified HNP 1-3 decreased the viability at 10 and 15 µg/ml and commercial HNP 1-3 at 15 µg/ml concentrations. Following to the purified HNP1-3 treatment, the percentage of Annexin V positive population and caspase-3 activity were significantly increased compared to control (p = 0.000 and p = 0.001, respectively) and commercial HNP1-3 (p = 0.034 and p = 0.018, respectively). SIGNIFICANCE: Results indicated the anticancer activity of HNP1-3 which can be used as future chemotherapeutic drugs. Furthermore, leukofilters can be considered as economic source for purifying these peptides.

Prevalence of Aminoglycoside Resistance Genes in Enterococcus Strains in Kermanshah, Iran.

BACKGROUND: This study aimed to investigate the occurrence of aminoglycoside resistance and the prevalence of the resistance-modifying enzyme genes, ant(3")-III, ant(6')-Ia, aac(6')-Ie-aph(2")-Ia, and aph(2')-Id, in Enterococcus strains isolated in Kermanshah Province, west of Iran. METHODS: In this cross-sectional study, 108 enterococcal isolates from urine, wound, blood, and cerebrospinal fluid samples were collected. The Enterococcus species were recognized by standard phenotypic/biochemical tests. The antimicrobial resistance forms were detected using a disc diffusion method. Polymerase chain reaction was designed to identify aminoglycoside resistance genes, including ant(3")-III, ant(6')-Ia, aac(6')-Ie-aph(2")-Ia, and aph(2')-Id. RESULTS: Totally, 108 strains with a final diagnosis of Enterococcus were extracted from 84 (77.8%) urine, 14 (13%) wound, 6 (5.6%) blood, and 4 (3.7%) cerebrospinal fluid samples. Among the 108 Enterococcus specimens, 94 (87%) cases were Enterococcus faecalis and 14 (13%) were Enterococcus faecium. The highest frequency of resistance was observed for erythromycin (88.9%), while the lowest was found for streptomycin (44.4%). The frequency of high-level gentamicin resistance was 42.2%. Among the identified specimens, 42.6% contained the aac(6')-Ie-aph(2")-I gene, 20.4% contained the ant(6')-Ia gene, and 15.7% contained the ant(3")-III gene. A significant correlation was found between phenotypic gentamicin resistance and the presence of the aminoglycoside resistance genes (P<0.05). CONCLUSION: This study showed the high resistance of Enterococcus strains isolated from hospital samples. Compared with the previous studies, the strains isolated in our study showed a higher percentage of resistance to aminoglycosides.

Evaluation of the Sensitivity and Specificity of Use of Glucose and pH for Bacterial Screening of Platelet Concentrates Compared to the Bact/Alert.

Bacterial contamination of blood components is the major infectious risk in transfusion medicine. Since platelets should be stored at room temperature that makes them an excellent growth medium for bacteria; it is mentioned as a major problem in transfusion medicine. Transfusion risk of a bacterial contaminated platelet concentrate is higher than viral pathogen such as HIV, HBV, HCV and HTLV. The objective of this study was to evaluation of the sensitivity and specificity of use of glucose and pH for bacterial screening of platelet concentrates compared to the Bact/Alert. 1332 platelet concentrates were screened by the Bact/Alert system for aerobic and anaerobic bacterial contamination. Bacterial contamination was also evaluated by using urine reagent strips (Multistix10 SG Bayer) and culture methods. Moreover PH screening with a pH meter (Metrohm 744 Swiss) and glucose was also used for detection of bacterial contamination. The rate of bacterial contamination detected by the Bact/Alert system in platelet concentrates was 25 in 1332 (1.9 %). It contained 15 (1.1 %) for aerobic bacteria and 10 (.8 %) for anaerobic bacteria. 226 of 1332 were considered as containing bacteria by using urine reagent strips. Six of the 226 units were also positive by the Bact/Alert system. Three of those units were culture positive for aerobic bacteria and three for anaerobic. The result of platelet concentrates that underwent pH screening by use of pH meter and a pH portion of urine reagent strips was the same. The sensitivity and specificity of considering glucose alone for detection of bacterial contamination were 12 and 98 % respectively. For pH alone, these were 24 and 83 %. For glucose and/or pH, these were 24 and 83 %; and for combination of glucose and pH, these were 12 and 98 %. Our results showed use of glucose/pH strips would improve the safety of blood products and should be encouraged.

ASXL1 and JAK2V617F gene mutation screening in Iranian patients with chronic myeloid leukemia.

AIM: In recent years, a few cases of chronic myeloid leukemia (CML) have been reported with both BCR-ABL and JAK2V617F mutations. Moreover, mutations in the additional sex comb-like 1 (ASXL1) gene were recently shown in various myeloid malignancies.There were no previous studies investigating the incidence of the ASXL1 and JAK2V617F mutations in Iranian patients with CML. Consequently, this study focuses on the analysis of these mutations in patients with CML. METHODS: In total, 66 patients with a clinical diagnosis of CML were examined at the time of diagnosis. Thirty healthy subjects were checked as controls. Exon 12 of ASXL1 was amplified from genomic DNA and bidirectionally sequenced. We also performed JAK2V617F screening by amplification refractory mutation system-polymerase chain reaction and sequencing. RESULTS: Mutations in the ASXL1 gene were found in five out of 66 CML patients (7.6%). We identified a novel variant (c.1968G > A, p.Asp656Asn) in one of the patients that has not been reported before. We also identified BCR-ABL and JAK2V617F mutations simultaneously in four patients (6%). CONCLUSION: Our demonstration of ASXL1 mutation, a putative tumor suppressor gene, represents an important molecular abnormality in CML. We also showed that concomitant detection of BCR-ABL and JAK2V617F mutations has a relatively high incidence in Iranian patients.

JAK2V617F Allele Burden Measurement in Peripheral Blood of Iranian Patients with Myeloproliferative Neoplasms and Effect of Hydroxyurea on JAK2V617F Allele Burden.

BACKGROUND: Myeloproliferative neoplasms (MPNs) are clonal malignant diseases that represent a group of conditions including polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). The aim of this study was to evaluate possible correlations between JAK2V617F allele burden and clinicohematologic characteristics in Iranian patients with MPNs. We also aimed at determining the correlation between JAK2V617F allele burden and use of cyto reductive treatment (hydroxyurea). MATERIALS AND METHODS: We performed ARMS-PCR for all MPNs samples and subsequently performed real-time quantitative polymerase chain reaction (qRT-PCR) for JAK2V617F allele burden measurement using DNA from peripheral blood leukocytes. RESULTS: Two distinct groups of patients were examined at a single time point: group A (n=40; 20 PV, 20 ET) was examined at the time of diagnosis; group B (n=85; 40 PV, 30 ET and 15 PMF) while under treatment with hydroxyurea (HU). The median allele burden of the JAK2 V617F was 72% for PV and 49% for ET patients at the time of diagnosis (p=0.01). For patients with HU treatment, we determined the median JAK2V617F allele burden to be 43%, 40%, and 46.5 % in PV, ET and PMF patients; respectively. HU-treated PV patients had a significant lower %JAK2V617F than PV patients at the time of diagnosis (43% vs. 72%, p=0.005). In ET group, the relationship between the JAK2 V617F allele burden and leukocyte count was significant (p=0.02 and p=0.01 in untreated and treated patients, respectively). CONCLUSIONS: Our results showed that patients with PV have a higher JAK2V617F allele burden. Moreover, our study demonstrated that the JAK2V617F allele burden correlates with clinical features in ET group. We also showed hydroxyurea can affect the JAK2V617F allele burden in PV patients.

RBC alloimmunization and double alloantibodies in thalassemic patients.

PURPOSE: Alloimmunization is a common consequence of chronic blood transfusion. Double alloantibody production may complicate the condition of such patients especially for finding matched blood. In this study, we evaluated the frequency of alloantibodies in thalassemic patients with previous history of transfusion reactions. SAMPLES AND METHODS: This study was performed on 441 multiply transfused thalassemia patients Antibody screening test was carried out using three cell-panel by gel method. Positive patients were followed up for antibody identification using 11-cell panel. Direct combs' test was performed to detect auto antibodies. RESULTS: In a total of 441 cases (362 thalassemia major and 79 intermedia), 234 were males (53.1%) and 207 females (46.9%); mean age 22 years, range 3-61 years. Alloimmunization was detected in 50(11.3%) patients, including 37(74%) patients with one alloantibody, 8(16%) with two antibodies, 4(8%) patients with unknown antibodies and one patient (2%) with autoantibody. The most common alloantibodies were anti-Rh antibodies (-E/e/C/c/Cw) (26%), anti-K (28%), anti-D (16%), and anti-Colton (4%). Double antibodies were detected in eight out of 50 patients, including: Anti-D+anti-C (8%), anti-D+anti-E (2%), anti-Kell+anti-D (2%), and anti-Kell+KPa (2%). A significant association was observed between the transfusion reaction history and the alloantibody detection results (p < 0.05). CONCLUSION: Antibody production against RBC antigens makes hard condition in regular blood transfusion. Double antibodies production may more complicate this situation. Thus, it is advisable to phenotype patients and matches the red cells in multiply transfused thalassemia patients.