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Recipient kidney damage after leukocyte transfer from inbred mice with renal ischemia-reperfusion injury
Kadkhodaee M, Khastar H, Seifi B, Najafi A, Delavari F,
Volume 70, Issue 2 (5-2012)
Abstract

Background: In a recent study, we were able to demonstrate a role for leukocyte transfer in the induction of liver damage in recipient mice after induction of IR (60 min of bilateral renal artery occlusion and 3 hrs reperfusion) injury in donors. The present study investigates the role of leukocyte transfer in the induction of kidney damage in recipient mice after induction of renal IR injury in donors.

Methods: Mice were divided into two sham and renal IR groups. After anesthesia, leukocytes were isolated from blood and were transferred to the two recipient groups: the intact recipient mice received leukocytes from the sham donor group (Sham recipient) and the intact recipient mice that received leukocytes from IR donor group (IR recipient). After 24 hrs, the recipient mice were anesthetized and blood samples and renal tissues were collected.

Results: Renal malondialdehyde (MDA) increased and glutathione and superoxide dismutase (SOD) decreased significantly in IR recipient group in comparison to sham recipient group. Although renal function tests, including BUN and plasma creatinine were significantly different between IR donor and sham donor groups, but they were not significantly different in two recipient groups. Renal tissues in IR donor group showed extensive damage compared to sham group, but in IR recipients' kidneys, they were different from IR donor tissues despite being different from their respective sham group.

Conclusion: These findings are suggestive of implication of leukocytes in renal tissue damage and oxidative stress after renal IR injury.


Leukocytes and transfusion related adverse events: the effects of leuko-reduction process in the prevention of adverse reactions resulted from the transfusion of blood components: review article
Ehteramolsadat Hosseini , Amin Shahbaz Ghasabeh , Mehran Ghasemzadeh ,
Volume 75, Issue 2 (5-2017)
Abstract

Blood transfusion is commonly implemented to manage life and health-threatening conditions on a rapid and short-term basis. Over the years, ongoing technical advances have dramatically improved transfusion medicine to provide more safety and effectiveness. However, transfusion is still complicated with different adverse events that mainly induced by the presence of allogeneic leukocytes in the blood products. Several lines of evidence have shown that leukocytes in blood components are involved in the induction of febrile nonhemolytic transfusion reactions (FNHTRs), HLA alloimmunization and platelet refractoriness as well as the increased risk of the infectious diseases transmitted by leukotropic viruses including cytomegalovirus (CMV), human T-lymphotropic virus (HTLV)-I/II and Epstein-Barr virus (EBV). During current decades, introducing various leuko-reduction techniques have shown to be associated with less transfusion related adverse events and improved clinical outcomes. The lower incidence and severity of febrile transfusion reactions; reduced risk of transfusion related transmission of CMV or other leukocyte-associated infections, lowered incidence of alloimmune platelet refractoriness in addition to reducing risk of mortality and morbidity in patients are considered as clinical benefits of leuko-reduced products. Currently, by the use of 3rd and 4th generation of filters, the highest levels of leukoreduction in blood components have been achieved. Filtration techniques have also the advantages of being performed shortly after preparation of components (pre-storage) or post-storage even at the patient’s bedside. However, it seems that pre-storage depletion of leukocytes provides better protection than post-storage techniques due to the elimination of leukocyte-derived cytokines effects which are increasingly released during storage. Particularly in platelet products, the earlier depletion of leukocyte also favors less platelet-induced leukocyte activation which may be triggered by the interaction between either activated platelets or their released chemokines and residual leukocytes during storage. Despite the benefits attributed to leukoreduction of blood components, the global use of leukoreduced products is commonly hampered by its high cost especially in developing countries in which leukoreduction of blood components is usually limited to some patients with special conditions. In this review, after briefly introducing of some transfusion adverse events that are attributed to allogeneic leukocytes existed in blood products, the effects of leukoreduction process in the attenuation of these events will be discussed.


A multi-step centrifuge method for extraction of leucocytes and preparation of Tc-HMPAO labeled leucocytes
Nasim Vahidfar, Mahdieh Parvizi, Marzyehsadat Peyman, Hana Safar, Saeed Farzenehfar , Mehrshad Abbasi,
Volume 80, Issue 2 (5-2022)
Abstract
Background: Labeled leucocytes could be used for localization of infection foci after surgeries or in inflammatory diseases including inflammatory bowel diseases. Extraction of leucocytes needs 10% Hetastarch which is not available in Iran. We provide a method employing multiple centrifuges to extract and label leucocytes with Tc-HMPAO.
Methods: The study was conducted from April to June 2018 in the Nuclear Medicine Unit of Valiasr Hospital. Leucocytes were extracted from a 60 ml blood sample anticoagulated with Acid-citrate-dextrose through four-step centrifugation as below: 1-whole blood was centrifuged at 1k cycle per minute (CPM) for eight minutes to precipitate red blood cells (RBC). Supernatant including RBC free plasma, WBC, and platelet was extracted for the next step. 2-WBC was precipitated at 1.8k CPM for five minutes and platelet-rich plasma (PRP) as supernatant. 3- PRP was centrifuged at 3k for five minutes and cell-free plasma (CFP) was extracted as supernatant, and 4- precipitate WBS at step two was washed with saline and centrifuged at 0.5k CPM to achieve washed WBC. Then the leucocytes were labeled with 40 mCi Tc-HMPAO through 15 minute incubation at 37-38 degrees centigrade. The extra free pertechnetate was eliminated using two additional centrifugation steps as follows: 1-0.5k CPM for five minutes to dispense free pertechnetate, and 2-0.5 for five minutes to achieve high radiochemical purity labeled WBC. Finally, the labeled WBC was re-suspended in CFP and reinjected to the patient. Imaging at 2-4 hours was done. The pathology and imaging of labeled WBC distribution are reported
Results: No RBC was detected in microscopy. The majority of the leucocytes were lymphocytes with rare accompanying platelets. The radiolabeling efficiency of the procedure was higher than 40%. The viability test indicated more than 80% of viable cells. The radiochemical purity of the final product was more than 95%. Two to four hours after injection, low background images were acquired. The liver and spleen were target organs with low-grade urinary, thyroid, and GI activity.
Conclusion: Employing multi-stage centrifugation, Tc-HMPAO labeled leucocyte scan could be efficiently performed.

Assessment of mitochondrial DNA copy number in peripheral blood leukocyte of opiate abusers and healthy individuals
Kouros Divsalar, Sara Hesami, Majid Mahmoodi, Navidreza Giahi, Fatemeh Divsalar , Mohammad Pour-Ranjbar , Amin Honarmand,
Volume 80, Issue 7 (10-2022)
Abstract
Background: Based on the studies, variation in the mitochondrial DNA (mtDNA) copy number in peripheral blood leukocytes is associated with increased susceptibility to diseases including cancer. Opiate abusers are at high risk for diseases. In this study, we measured the mtDNA copy number in peripheral blood leukocytes in a group of opiate abusers compared with those in healthy individuals.
Methods: In a case/control study, three groups were selected consisting of 32 opium abusers, 24 heroin addicts and 25 healthy individuals. The amount of 5 ml of whole blood was collected from each individual who participated in the study and stored at -20 centigrade. The sample collection was performed from November 2018 to February 2020. Case groups were recruited from the Methadone maintenance therapy center. Contro group had no history of drug use and cigarette smoking. DNA was extracted from the whole blood samples using the salting out method. The DNA from a mitochondrial gene, dehydrogenase subunit1 (-ND1 gene) and a nuclear gene, human globulin (HGB gene), were quantified by a real-time PCR-based method to measure the relative mtDNA copy number of each group number.
Results: There was no significant difference in demographic characterization between the three study groups, opium abusers, heroin addicts and healthy individuals. We found that opium users had a higher mean of mtDNA copy number than those in the healthy control group (P=0.11). Heroin addicts had also higher mean of mtDNA copy number than those in healthy group (P=0.21). The mean mtDNA copy number in opium abusers was higher than that in heroin addicts (P=0.22), although the difference was not statistically significant.
Conclusion: The results of this study indicated that mtDNA copy number increased in a group of opiate abusers. Considering that alteration of mtDNA copy number is associated with increased susceptibility to several diseases including cancer, further research on mtDNA copy number with a high number of volunteers of opiate addicts may clear the effect of opiate abuse on the human genome.

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