Although the use of cryoprotectant dimethyl sulfoxide (DMSO) is the gold standard in cryopreservation of hematopoietic stem cells, it is well known that it has a negative effect on cell viability. ...The aim of this prospective study was to examine how the length of post-thaw exposure to DMSO affects the cell viability and stability of peripheral blood stem cell (PBSC) samples. Additionally, the effects of donor type and pre-cryopreservation storage time on post-thaw viability during the stability study were evaluated. In 30 autologous and 30 allogeneic PBSC samples viable CD34+, CD14+, CD19+, CD16+/56+, and CD3+ cells were determined immediately after thawing, and one-and three-hours post-thaw.
Analysis of the absolute count of viable cells in thawed samples showed a significant difference between all measurement points for CD34+ (
< 0.001), CD14+ (
< 0.001), and CD19+ cells (
< 0.001). No significant differences were observed for post-thaw stability of allogeneic samples analysed between products stored before cryopreservation ≥ 24 hours (N = 20), and those stored < 24 hours (N = 10), except for viable CD3+/CD4+ cells after three hours post-thaw (
= 0.028). In conclusion, DMSO had different effects on leukocyte subpopulations in cryopre-served PBSC samples. The type of donors and the length of storage before cryopreservation did not affect the post-thaw stability of cryopreserved PBSC samples.
Introduction
CD34+ hematopoietic stem cell (HSC) enumeration by cell flow cytometry is routinely used in clinical laboratories for monitoring of HSC mobilization into peripheral blood and assessment ...of the quality of HSC products. The modified ISHAGE protocol is the most often used procedure for determination of CD34+ cells using flow cytometry. The aim of this study was to evaluate BD Enumeration stem cell kit on flow cytometer BD facscanto II, using facscanto clinical and facsdiva softwares.
Methods
Validation study included determination of within‐run and between‐run precision, trueness (bias), comparison of the test results analyzed on facscanto clinical and facsdiva softwares, assessment of linearity, specimen stability, and carryover.
Results
For between‐run precision, coefficients of variation (CVs) were all <10%, except for low control level on facsdiva software. CVs for within‐run precision were <10%, except for high absolute count of CD34+ cells on facsdiva software. Comparison of data showed no statistically significant differences between facscanto clinical and facsdiva software (Spearman's rank correlation coefficients were .993 for % of CD34+ cells and 0.983 for absolute count of CD34+ cells). In linearity study, bias for all dilutions was < 20%, and carryover assessment cannot be considered significant on both softwares. There was a statistically significant difference (P = .044) in absolute count of CD34+ cells after 24 hours of storage, when using facscanto clinical software.
Conclusion
BD Stem Cell Enumeration Kit can be used in routine laboratory work on BD FACSCanto II instrument, whereas facscanto clinical and facsdiva software were used for acquisition and data analysis.
SAŽETAK
Autoimunosna hemolitička anemija je rijetka bolest imunosnog sustava koju karakterizira hemoliza vlastitih eritrocita uzrokovana autoprotutijelima i/ili aktiviranim komponentama komplementa. ...Ovisno o osnovnoj bolesti, autoimunosna hemolitička anemija može biti primarna ili sekundarna, a prema serološkim karakteristikama autoprotutijela dijeli se na toplu, hladnu i miješanu. Dijagnoza bolesti postavlja se na temelju pozitivnog rezultata direktnoga antiglobulinskog testa, anemije i prisutnih laboratorijskih pokazatelja hemolize. U vrlo rijetkim slučajevima mogući su oblici autoimunosnih hemolitičkih anemija s negativnim direktnim antiglobulinskim testom. Uz preciznu imunohematološku dijagnostiku potrebnu za razlikovanje pojedinih vrsta autoimunosnih anemija, za pravilno liječenje nužna je i dijagnostika osnovne bolesti. U liječenju toplih autoimunosnih hemolitičkih anemija najčešće se primjenjuju kortikosteroidi, a u težim slučajevima i rituksimab. Kod hladnih autoimunosnih hemolitičkih anemija važno je utopliti bolesnika i izbjegavanje hladnoće, a medikamentozno liječenje rituksimabom s bendamustinom ili bez njega potrebno je u težim slučajevima. U refraktornim slučajevima paroksizmalne hladne hemoglobinurije uz rituksimab se primjenjuju imunosupresivni lijekovi. Za miješanu autoimunosnu hemolitičku anemiju uz kortikosteroide se preporučuje rano primijeniti rituksimab. U liječenju simptomatske anemije primjenjuje se transfuzijsko liječenje, a za liječenje životno ugroženog bolesnika moguće je primijeniti intravenske imunoglobuline, plazmaferezu, a u rijetkim slučajevima retikulocitopenije i eritropoetin. Novi lijekovi, koji su trenutno u kliničkim istraživanjima, obećavajući su za liječenje teških oblika autoimunosnih hemolitičkih anemija, kao što su komplementom posredovane autoimunosne hemolitičke anemije ili refraktorni oblici hladnih autoimunosnih hemolitičkih anemija. Za praćenje terapijskog učinka autoimunosnih hemolitičkih anemija važno je poznavati kriterije kojima se definira odgovor na terapiju i ishod bolesti. Cilj ovoga preglednog rada jest prikazati aktualna saznanja o dijagnostici i liječenju te praćenju učinka terapije autoimunosnih hemolitičkih anemija.
SAŽETAK
UvodPandemija COVID-19 dovela je do izazova u liječenju bolesnika kojima je potrebna transplantacija krvotvornih matičnih stanica (KMS). Porastom broja oboljelih širom svijeta smanjila se ...dostupnost KMS-a zbog infekcije darivatelja i ograničenja u međunarodnom prometu. Sukladno tomu, uveden je niz mjera u cilju zaštite bolesnika i darivatelja te osiguravanja dostupnosti transplantata tijekom pandemije. Cilj rada bio je prikazati utjecaj pandemije COVID-19 na prikupljanje alogeničnih KMS-a u KBC-u Zagreb.
MetodeProvedena je retrospektivna analiza podataka za razdoblje od 1. ožujka 2020. do 30. lipnja 2021. Podatci su prikupljeni iz bolničkoga informacijskog sustava te zapisnika sa sastanaka Povjerenstva za liječenje transplantacijom KMS-a KBC-a Zagreb.
RezultatiU navedenom razdoblju prvenstveno su korištene KMS iz periferne krvi, osim u slučaju snažne indikacije za primjenu koštane srži (KS). Pripravci alogeničnih KMS-a su prije početka kondicioniranja krioprezervirani kako bi se osigurala raspoloživost prema planu liječenja. Kod 13 bolesnika se odustalo od prikupljanja KMS-a jer su dobili COVID-19. Alogenične KMS su prikupljene od srodnih i nesrodnih darivatelja iz Hrvatskog registra dobrovoljnih darivatelja KMS-a i Svjetskog udruženja darivatelja koštane srži za ukupno 135 bolesnika. Koštana srž je prikupljena za 12,5% bolesnika i svi pripravci su transplantirani. Šesnaest bolesnika je transplantirano s perifernim KMS umjesto KS. Transplantirano je 94,1% pripravaka perifernih KMS-a, od kojih je 17 transplantirano s odgodom zbog infekcije darivatelja i/ili bolesnika bolešću COVID-19 prije ili nakon prikupljanja KMS-a. Od ukupno 118 transplantata perifernih KMS-a, 100 (84,7%) ih je krioprezervirano u 540 vrećica. Sedam (5,9%) krioprezerviranih transplantata perifernih KMS-a nisu infundirani zbog progresije bolesti (5), infekcije primatelja bolešću COVID-19 (1) i loše vijabilnosti stanica (1).
ZaključakPandemija COVID-19 nepovoljno je utjecala na prikupljanje i transplantaciju KMS-a uz brojne organizacijske i logističke izazove. Krioprezervacija alogeničnih KMS-a omogućila je uspješno odvijanje transplantacijskog liječenja, iako je nažalost praćena rizikom da neki od tih pripravaka kasnije ne budu infundirani, što izlaže darivatelje riziku nepotrebnog prikupljanja i povećava troškove liječenja.
Donated human milk is the best possible alternative when mother's own milk is not available. The aim of this study is to investigate whether there are differences in the milk donation volumes and ...microbiological quality of donated milk depending on human milk donors (HMDs) characteristics.
We analyzed data on the HMDs who donated milk in the first three years of work of human milk bank (HMB) - November 2019 to January 2023. The data on the volume of donated milk in L and suitable microbiological quality assessed by the number and isolated species of bacteria were collected from questionnaires filled out by HMDs and documentation administered by HMB employees and are presented using descriptive and comparative statistics.
Two hundred HMDs were included in this study. The majority of them are between 26 and 35 years of age, reside in capital city or the surrounding county, have given birth to a full-term child vaginally, and express surplus milk through a breast pump. Donor mothers of preterm born infants (14.5 %) donated greater quantities, there is statistically significant difference in the median of volume of milk donated (9.6 vs. 6.4, p=0.026). Milk expressed manually shows better results in microbiological quality (median percentage 100 vs. 82 vs. 100, p=0.040), while by comparing other characteristics of the donors, no difference was found between the groups.
It is important to be aware of the characteristics of previous HMDs in order to direct the HMB future promotional and educational activities.
Fetal/neonatal alloimmune thrombocytopenia (FNAIT) is a rare life‐threatening disorder, leading to severe thrombocytopenia and potentially bleeding, with intracranial haemorrhage (ICH) being the most ...serious complication. We report on a FNAIT case with fourth‐degree ICH that arose due to antibodies against human platelet antigen (HPA)‐1b. The male infant, born to an otherwise healthy mother, presented with severe signs of ICH soon after delivery. Since only moderate thrombocytopenia was noted and there were no active signs of bleeding, the infant did not receive intravenous immunoglobulins (IVIg) or platelet transfusion. Spontaneous recovery of platelets was noted on the eighth day of life, but permanent neurological impairment remained as a consequence of ICH. We report the results of HPA and human leukocyte antigen (HLA) antibodies in the mother's and the infant's sera, the family's HPA genotype and the mother's HLA genotype, and summarise previously described cases of FNAIT due to anti‐HPA‐1b antibodies in the literature. FNAIT with severe ICH due to anti‐HPA‐1b antibodies is rarely diagnosed. An association between HLA genes and sensitization to HPA‐1b antibodies was not demonstrated. The severity of FNAIT and the occurrence of ICH is often difficult to predict. In this case, the infant presented with moderate thrombocytopenia and ICH, with subsequent permanent consequences.
Background
Patients with hematological diseases are polytransfused and often immunocompromised, therefore susceptible to transfusion reactions (TR). This study aims to document the incidence of TRs ...in adult hematological patients and assess the effect of changes in the production of blood components and transfusion practice on their occurrence.
Study design and methods
Retrospective observational analysis of TRs reported from 1993 to 2019 was performed. For the analysis of the effect of changes on the incidence of TRs, the evaluated time was divided into two periods: the 1st period before the introduction of changes in production, when leukoreduced blood components were used only selectively, and the 2nd period, when semi‐automated method of production and universal leukoreduction was introduced.
Results
The decrease in the incidence of TRs was observed for both red blood cell (RBC) and platelet concentrate (PC) transfusions in the 2nd period. Since platelet additive solution has been used, a further decrease in the incidence was reported. The decrease in incidence was also observed for delayed hemolytic/serological transfusion reactions and for transfusion‐transmitted bacterial infections. Four cases of incorrect blood transfusions were uniquely related to the hematological patients, caused by antigen loss and transfusion ordering after ABO‐incompatible hematopoietic stem cell transplantation.
Discussion
Our results provided evidence that the introduction of tools offered by modern transfusion medicine: universal leukodepletion, plasma replacement with additive solutions, sensitive laboratory techniques, prophylactic antigen matching policy, informatization, and automatization, decreased the incidence of TRs and improved transfusion safety.
Alloimmunization is a known risk of transfusion therapy caused by exposure to foreign RBC antigens. However, alloimmunization is not observed in all transfused patients. Human leukocyte antigen (HLA) ...molecules may contribute to the recognition and presentation of foreign antigens and to the potency of immune responses that result in the production of antibodies. The aim of this study was to determine the association of HLA-DR and HLA-DQ polymorphisms with alloimunization to Fy
antigen in Croatian patients.
The study was conducted on 70 alloimmunized patients to Fy
antigen and two control groups: 165 healthy Croatian individuals (Control 1) and 45 Fy
antigen-negative nonimmunized patients exposed to Fy
antigen (Control 2). Phenotype frequencies for HLA-DRB1 and HLA-DQB1 alleles were compared between the cases and control groups.
Statistically significant differences in phenotype frequencies between cases and controls were found for DRB1*04 (odds ratios ORs, 10.5 and 18.7 for Control 1 and Control 2, respectively), DRB1*15 (ORs, 8.0 and 6.9), and DQB1*02 alleles (ORs, 0.2 and 0.03); and DRB1*04-DQB1*03:01 (ORs, 7.9 and 17.6), DRB1*04-DQB1*03:02 (ORs, 5.5 and 7.6), DRB1*15-DQB1*06:02 (ORs, 7.3 and 5.5), DRB1*03-DQB1*02:01 (OR, 0.1), and DRB1*07-DQB1*02:02 (OR, 0.3) haplotypes.
Several HLA-DRB1 and HLA-DQB1 alleles and haplotypes were proved to contribute to and protect from alloimmunization to Fy
antigens. Alleles DRB1*04 and DRB1*15, as well as haplotypes DRB1*04-DQB1*03:02 and DRB1*15-DQB1*06:02 can be considered as risk factors, while allele DQB1*02 and haplotype DRB1*03-DQB1*02:01 have a protective role in Fy
alloimmunization.
•In clinical practice, a substantial proportion of platelet transfusions are routinely administered outside the guidelines.•Unnecessary platelet transfusions are an unjustified extra burden on a ...scarce healthcare resource and may also be detrimental to the patients.•One-third of all PLT transfusions were inappropriate mainly because of transfusions above the recommended threshold and unjustified double units transfusions.
Hematology patients are intensive platelet users. In clinical practice, a substantial proportion of platelet (PLT) transfusions are routinely administered outside the guidelines despite compelling evidence for recommendations. Those unnecessary PLT transfusions are an unjustified extra burden on a scarce healthcare resource and may also be detrimental to the patients. This study aims to evaluate indications and assess the appropriateness of PLT transfusion, as well as to identify common discrepancies and propose modalities for better compliance with guidelines.
The audit of all PLT orders for adult hematological inpatients was conducted over 2 months. The assessment was performed using guidelines for PLT transfusion. Patient demographic, clinical, and transfusion data were collected from hospital electronic medical records.
Based on 286 PLT orders, 344 PCs were transfused to 67 patients: 235 (82.2%) prophylactical due to low PLT count, 34 (11.9%) preprocedural and 17 (5.9%) therapeutic. Overall, 105 (36.77%) PLT transfusions were inappropriate: 78 (33.2%) of all prophylactic PLT transfusions due to low PLT count, 17 (50%) off all preprocedural and 10 (58.8%) of all therapeutical transfusion. The major reason for PLT transfusion inappropriateness was transfusion above the recommended threshold. Double units of PCs were transfused in 36.7% of all PLT transfusions and 32.4% of them were considered inappropriate.
Our audit of PLT transfusion practice found a large proportion of inappropriate PLT transfusions. Based on the most common deviations from the guidelines a variety of targeted measures for improvement are proposed.
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