Metahemoglobinemia y hemólisis intravascular aguda después de una intoxicación por naftaleno en un paciente pediátrico / Methemoglobinemia and acute intravascular hemolysis after naphthalene poisoning in a pediatric patient

La intoxicación por naftaleno es poco frecuente en los niños. Es causada por la ingesta, la inhalación o el contacto con la piel de sustancias que contienen naftaleno. Los pacientes suelen tener orina de color marrón oscuro, diarrea acuosa y vómito bilioso. Los signos incluyen fiebre, taquicardia, hipotensión y valores bajos en la oximetría de pulso, incluso con oxigenoterapia. Los análisis de sangre detectan anemia hemolítica, metahemoglobinemia, insuficiencia renal e hiperbilirrubinemia. Además del tratamiento sintomático, se hacen transfusiones de eritrocitos y se les administran ácido ascórbico, azul de metileno y N-acetilcisteína. En este artículo, presentamos el caso de un paciente masculino de 23 meses de edad con metahemoglobinemia y hemólisis intravascular aguda que recibió atención en la unidad de cuidados intensivos durante cinco días por intoxicación por naftaleno. Si bien la intoxicación por naftaleno es muy poco frecuente, tiene consecuencias mortales y se debe ejercer precaución con su uso y venta.

Poisoning by naphthalene is uncommon in children. It is a type of poisoning brought on by ingesting, inhaling, or coming into touch with naphthalene-containing substances on the skin. Patients typically present with an initial onset of dark brown urine, watery diarrhea, and bile vomit. The signs include fever, tachycardia, hypotension, and low pulse oximetry readings even with oxygen support. Hemolytic anemia, methemoglobinemia, renal failure, and hyperbilirubinemia are all detected in blood tests. Erythrocyte transfusion, ascorbic acid, methylene blue, and N-acetylcysteine (NAC) therapies are provided to inpatients in addition to symptomatic treatment. We present a 23-month-old male patient who developed methemoglobinemia and acute intravascular hemolysis, who was followed up in the intensive care unit for five days due to naphthalene intoxication. Although naphthalene poisoning is very rare, it should be known that it has fatal consequences, and more care should be taken in its use and sale.

Long-term outcome of transcatheter repair of paravalvular leak post surgical mitral valve replacement / 中华心血管病杂志

Objective: To explore the long-term clinical efficacy of transcatheter repair of mitral paravalvular leak (PVL) post surgical mitral valve replacement. Methods: This study is a retrospective study. Patients who completed transcatheter repair of paravalvular leak after surgical mitral valve replacement at Shanghai Chest Hospital, Shanghai Jiaotong University School of Medicine from March 2010 to December 2018 were included. Technical success was defined as the occluder being stably implanted in the paravalvular leak site without affecting the function of the mitral valve and surrounding tissues; and there were no intervention-related complications, such as new hemolysis or aggravated hemolysis, and echocardiography confirmed mitral paravalvular regurgitation reduced by more than 1 grade. Patients were followed up at 30 days, 1, and 3 years after the intervention. The main endpoints were all-cause death and re-surgery due to interventional failure or serious complications. The occurrence of occluder-mediated hemolysis and chronic renal insufficiency was recorded, and patients were monitored with echocardiography during follow up. Results: A total of 75 patients were included, aged (54.3±22.9) years old, and 38 patients were males. All patients had decreased cardiac function and/or hemolysis before intervention. Procedural success was achieved in 54 patients (72.0%). Incidence of device-mediated hemolysis was 18.7% (14/75). During the follow-up period, all-cause death occurred in 7 patients (9.3%), and 3 were cardiac deaths.The 3-year event-free survival rate was 81.3% (61/75). The need for cardiac surgery was 9.3% (7/75): 3 cases due to severe device-mediated hemolysis, 2 cases due to prosthetic valve failure and 2 cases due to moderate to severe residual regurgitation. The echocardiography follow-up results showed that the position of the occluder was stable, there was no impact on the artificial valve function and surrounding structures, and the residual regurgitation was stable without progressive increase in event-free patients. Compared with pre-intervention, the left ventricular end systolic diameter ((33.9±7.4)mm vs. (38.3±8.9) mm, P=0.036), end diastolic diameter ((53.7±8.3) mm vs. (58.4±9.1) mm, P=0.045) and left atrial diameter (59.3 (44.5, 90.7) mm vs. 64.3 (44.8, 96.6) mm, P=0.049) were significantly reduced, pulmonary artery systolic pressure was also significantly decreased ((36.5±15.8) mmHg vs. (46.3±14.9) mmHg, P=0.022, 1 mmHg=0.133 kPa). There was no significant difference between 3 years and 1 year after transcatheter repair of mitral paravalvular leak post surgical mitral valve replacement (all P>0.05). Conclusion: Transcatheter repair of mitral paravalvular leak post surgical mitral valve replacement is an effective treatment option in selective patients.

Retrospective Analysis of Irregular Antibodies Causing Hemolytic Disease of the Fetus and Newborn in Jiangxi Province / 中国实验血液学杂志

OBJECTIVE@#To analyze the characteristics of antibody-specific distribution, laboratory detection results of hemolytic disease of the fetus and neonatal(HDFN) caused by irregular blood group antibodies other than ABO, and its correlation with the clinical situation.@*METHODS@#The non-ABO-HDFN cases in our hospital from October 2012 to December 2021 were selected as the research objects, and the cases diagnosed with ABO-HDFN in the same period were randomly selected as the control group, and the data of antibody specific distribution, total bilirubin, direct antibodies, maternal history, age of the children, the presence or absence of combined ABO-HDFN, and whether to exchange/transfuse blood were retrospectively analyzed. The characteristics of non-ABO-HDFN in Jiangxi province were analyzed.@*RESULTS@#The detection rate of non-ABO-HDFN in Jiangxi province increased. Among 187 non ABO-HDFN cases, the highest percentage of Rh-HDFN was detected (94.6%). Compared with the control group of ABO-HDFN, the non-ABO-HDFN had higher mean integral value of direct antibody, higher peak total bilirubin, and longer duration. Anti-M-HDFN may have severe disease but the direct antibody weak positive/negative, it was easy missed in clinical and delayed the treatment. There is no correlation between the specificity of irregular antibodies, the sex of the child, the mother's previous childbirth history, the presence or absence of combined ABO-HDFN and the need for blood exchange/transfusion(P>0.05).@*CONCLUSION@#The irregular antibodies of causing non ABO-HDFN in Jiangxi area are mainly Rh blood group system, followed by MNS blood group system. Understanding the characteristics of HDFN disease, serological features and the correlation with clinical indexes will help to detect and treat non ABO-HDFN in time and reduce the risk of complications.

Clinical and genetic diagnosis for 26 paitents with hereditary spherocytosis / 中南大学学报(医学版)

OBJECTIVES@#Hereditary spherocytosis (HS) is the most common hereditary defect of the red cell membrane, mainly characterized by anemia, jaundice, and splenomegaly. Due to the atypical clinical manifestations and negative family history of some patients, as well as the low sensitivity and specificity of traditional laboratory examinations, it is easy for it to escape diagnosis or be misdiagnosed. At present, it has been confirmed that the mutation of ANK1, SPTB, SPTA1, SLC4A1 and EPB42 genes can cause the deletion of their corresponding coding proteins, and thus lead to the defect of erythrocyte membrane. This study aims to analyze the feasibility and clinical application value of HS gene diagnosis.@*METHODS@#Data of 26 patients from Hunan, China with HS admitted to the Department of Hematology, Second Xiangya Hospital of Central South University from January 2018 to September 2021 were retrospectively collected, and their clinical manifestations and results of laboratory examinations were analyzed. Next-generation sequencing (NGS) combined with Sanger sequencing were applied. The mutation of HS pathogenic gene and the variation of uridine diphosphate-glucuronosyl transferase 1 family polypeptide A1 (UGT1A1), a key enzyme in the regulation of bilirubin metabolism, were detected. The results of pathogenic gene variations were interpreted pathogenic gene variations in accordance with the Standards and guidelines for the interpretation of sequence variants published by the American College of Medical Genetics and Genomics (ACMG). The clinical characteristics of patients with different gene variants were analyzed, and the clinical diagnosis and genetic diagnosis were compared.@*RESULTS@#Among the 26 patients with HS, there were 23 cases of anemia, 25 cases of jaundice, 24 cases of splenomegaly, and 14 cases of cholelithiasis. There were 16 cases with family history and 10 cases without family history. The results of HS mutation test were positive in 25 cases and negative in 1 case. A total of 18 heterozygous mutations of HS pathogenic genes were detected in 19 families, among which 14 were pathogenic, 1 was likely pathogenic and 3 were of unknown significance. SPTB mutations (12) and ANK1 mutations (4) were the most common. The main variation types were nonsense mutation (9). There were no significant differences in peripheral blood cell parameters and hemolysis indicators between the SPTB mutant group and the ANK1 mutant group (all P>0.05). The rate of splenectomy in ANK1 mutation group was higher than that in SPTB mutation group, and the difference was statistically significant (χ2=6.970, P=0.014). There were no significant differences in peripheral blood cell parameters and hemolysis indicators among different mutation types (nonsense mutation, frameshift mutation, splice site mutation and missense mutation) (all P>0.05). Among the 18 clinically confirmedpatients, there were 17 cases whose diagnosis is consistent with the genetic diagnosis. Eight patients were clinically suspected, and all of them were confirmed by detection of HS gene mutation. Twenty-four patients with HS underwent UGT1A1 mutation detection, among which 5 patients carried UGT1A1 mutation resulting in a decrease in enzyme activity, and 19 patients had normal enzyme activity. The level of total bilirubin (TBIL) in the group with reduced enzyme activity was higher than that in the group with normal enzyme activity, and the difference was statistically significant (U=22, P=0.038).@*CONCLUSIONS@#Most patients with HS have anemia, jaundice and splenomegaly, often accompanied by cholelithiasis. SPTB and ANK1 mutations are the most common mutations in HS pathogenic genes among patients in Hunan, China, and there was no significant correlation between genotype and clinical phenotype. Genetic diagnosis is highly consistent with clinical diagnosis. The decrease of UGT1A1 enzyme activity can lead to the aggravation of jaundice in HS patients. Clinical combined gene diagnosis is beneficial for the rapid and precision diagnosis of HS. The detection of UGT1A1 enzyme activity related gene variation plays an important role in evaluation of HS jaundice.

Correlation Analysis of Hemolytic Transfusion Reaction Induced by Low Titer Antibody / 中国实验血液学杂志

OBJECTIVE@#To establish the diagnostic process of low titer blood group antibody in the occurrence of adverse reactions of hemolytic transfusion.@*METHODS@#Acid elusion test, enzyme method and PEG method were used for antibody identification. Combined with the patient's clinical symptoms and relevant inspection indexes, the irregular antibodies leading to hemolysis were detected.@*RESULTS@#The patient's irregular antibody screening was positive, and it was determined that there was anti-Lea antibody in the serum. After the transfusion reaction, the low titer anti-E antibody was detected by enhanced test. The patient's Rh typing was Ccee, while the transfused red blood cells were ccEE. The new and old samples of the patient were matched with the transfused red blood cells by PEG method, and the major were incompatible. The evidence of hemolytic transfusion reaction was found.@*CONCLUSION@#Antibodies with low titer in serum are not easy to be detected, which often lead to severe hemolytic transfusion reaction.

Analysis of frequency and molecular genetics of Jk (a-b-) phenotype among blood donors from Jining area / 中华医学遗传学杂志

OBJECTIVE@#To screen for Jk(a-b-) phenotype among blood donors from Jining area and explore its molecular basis to enrich the rare blood group bank for the region.@*METHODS@#The population who donated blood gratuitously at Jining Blood Center from July 2019 to January 2021 were selected as the study subjects. The Jk(a-b-) phenotype was screened with the 2 mol/L urea lysis method, and the result was confirmed by using classical serological methods. Exons 3 to 10 of the SLC14A1 gene and its flanking regions were subjected to Sanger sequencing.@*RESULTS@#Among 95 500 donors, urea hemolysis test has identified three without hemolysis, which was verified by serological method as the Jk(a-b-) phenotype and demonstrated no anti-Jk3 antibody. The frequency of the Jk(a-b-) phenotype in Jining area is therefore 0.0031%. Gene sequencing and haplotype analysis showed that the genotypes of the three samples were JK*02N.01/JK*02N.01, JK*02N.01/JK-02-230A and JK*02N.20/JK-02-230A, respectively.@*CONCLUSION@#The splicing variant of c.342-1G>A in intron 4, missense variants of c.230G>A in exon 4, and c.647_ 648delAC in exon 6 probably underlay the Jk(a-b-) phenotype in the local population, which is different from other regions in China. The c.230G>A variant was unreported previously.

Validation of blood components transport through a pneumatic tube system

ABSTRACT Introduction: Urgent blood component transfusions may be life-saving for patients in hemorrhagic shock. Measures to reduce the time taken to provide these transfusions, such as uncrossmatched transfusion or abbreviated testing, are available. However, transport time is still an additional delay and the use of a pneumatic tube system (PTS) may be an alternative to shorten the transport time of blood components. Objectives: To assess pneumatic tube system transportation of blood components based on a validation protocol. Methods: Pre- and post-transport quality control laboratory parameters, visual appearance, transport time and temperature of the packed red blood cells (RBCs), thawed fresh plasma (TFP), cryoprecipitate (CR), and platelet concentrate (PC) were evaluated. Parameters were compared between transport via pneumatic tube and courier. Results: A total of 23 units of RBCs, 50 units of TFP, 30 units of CR and ten units of PC were evaluated. No statistically significant differences were found between pre- and post-transport laboratory results. There was also no difference in laboratory parameters between transport modalities (PTS versus courier). All blood components transported matched regulatory requirements for quality criteria. The temperature during transport remained stable and the transport time via PTS was significantly shorter than the courier's transport time (p < 0.05). Conclusion: The PTS was considered a fast, safe and reliable means of transportation for blood components, also securing quality prerequisites.

Validación de un método analítico para cuantificación de ferritina en líquido cefalorraquídeo / Validation of an analytical method for the quantification of ferritin in cerebrospinal fluid

Resumen La ferritina es una proteína de gran tamaño que se encuentra fisiológicamente en el líquido cefalorraquídeo (LCR) en concentraciones de 2-10 ng/mL. Su elevación puede utilizarse como biomarcador en distintas condiciones patológicas. El procedimiento de validación tradicional para la medición en LCR no puede ser utilizado debido a la ausencia de controles y calibradores comerciales para esta matriz. El objetivo de este trabajo fue llevar a cabo una validación analítica de ferritina en LCR. Se realizaron ensayos de estimación de precisión y veracidad mediante el protocolo EP15-A3, linealidad por el protocolo EP6-A (ambos de la guía de la CLSI), recuperación, estabilidad, contaminación por arrastre, interferencia por hemólisis y bilirrubina y límite de detección (LoD). La ferritina en LCR en el autoanalizador DxI 800 de Beckman Coulter tuvo una performance intra e interensayo <3,7%, el ensayo denota linealidad en el intervalo de 2,1-547 ng/mL; se estableció estabilidad por un período de 5 días y la recuperación resultó ser aceptable. No se evidenció efecto de contaminación por arrastre ni interferencia por hemólisis hasta un rango entre 300-500 mg/dL de hemoglobina, ni interferencia por bilirrubina hasta una concentración de 16,0 mg/dL de bilirrubina total. El LoD fue de 0,4 ng/mL. Por medio de los ensayos realizados se logró validar la ferritina en LCR a partir de la utilización de pools de muestras, lo que pudo garantizar la confiabilidad y exactitud del método analítico.

Abstract Ferritin is a large protein physiologically present in the cerebrospinal fluid (CSF) in concentrations of 2-10 ng/mL. Its elevation can be used as a biomarker in several pathological conditions. The traditional validation procedure for measurement in CSF cannot be used due to the absence of commercial controls and calibrators for this matrix. The objective of the present study was to perform CSF ferritin analytical validation. Assays such as precision and accuracy estimation through the EP15-A3 protocol, linearity according to the EP6-A protocol (both from the CLSI guidelines), recovery, stability, carry-over, hemolysis and bilirubin interference and limit of detection (LoD) were conducted. Serum samples with different concentrations of ferritin were added to aliquots of a normal CSF pool. CSF ferritin in the Beckman Coulter DxI 800 had a <3.7% intra and inter-assay performance, the assay shows linearity in the 2.1 -547 ng/mL interval, stability was established for a 5-day period and the recovery was acceptable. There was neither carry-over effect or hemolysis interference up to a range of 300-500 mg/dL of hemoglobin, nor interference by bilirubin up to 16.0 mg/dL of total bilirubin. The LoD was 0.4 ng/mL. By means of the performed assays, CSF ferritin was validated by using sample pools, thereby ensuring the reliability and accuracy of the analytical method.

Resumo A ferritina é uma grande proteína fisiologicamente encontrada no líquido cefalorraquidiano (LCR) em concentrações de 2 a 10 ng/mL. Sua elevação pode ser usada como biomarcador em diferentes condições patológicas. O procedimento de validação tradicional para medição no LCR não pode ser usado devido à ausência de controles e calibradores comerciais para esta matriz. O objetivo deste estudo foi realizar uma validação analítica da ferritina no LCR. Foram realizados estudos de precisão e veracidade utilizando o protocolo EP15-A3, linearidade pelo protocolo EP6-A (ambos das diretrizes do CLSI), recuperação, estabilidade, contaminação transportada, interferência de hemólise e bilirrubina e limite de detecção (LoD). A ferritina no LCR no DxI 800 da Beckman Coulter teve um desempenho intra e inter-ensaio <3,7%, o ensaio denota linearidade na faixa de 2,1-547 ng/mL, a estabilidade foi estabelecida em um período de 5 dias e a recuperação foi considerado aceitável. Nenhum efeito de transporte ou interferência por hemólise foi evidenciado até um intervalo entre 300-500 mg/dL de hemoglobina, nem interferência pela bilirrubina até uma concentração de 16,0 mg/dL de bilirrubina total. O LoD foi de 0,4 ng/mL. Através dos testes realizados, a ferritina no LCR foi validada, com base no uso de pool de amostras, o que poderia garantir a confiabilidade e a acurácia do método analítico.

A review of the role of infections in the aetiology of haemolysis in patients with sickle cell diseases: pathogenesis, management, and prevention

Background: Sickle cell disease (SCD) is associated with chronic haemolysis, immuno-suppression and susceptibility to infections, which may trigger infection-associated haemolysis (IAH). SCD patients are vulnerable to anaemic effect of IAH due to vicious interaction between pre-existing 'inherited' chronic haemolysis and 'acquired' IAH. IAH in SCD manifests as febrile haemolytic crisis with clinical and laboratory features of severe anaemia or pancytopenia. Clinico-pathological perspectives of IAH in SCD are fragmented. This review presents a comprehensive but concise overview of pathogenesis, management and prevention of IAH in SCD. Methodology and results: Online literature search using search terms such as 'sickle cell disease, viral, bacterial, parasitic, fungal, infections, hyperhaemolytic crisis, haemophagocytic syndrome, severe anaemia, pancytopenia' in various combinations was done on PubMed/Medline, Google, Google-Scholar and Bing. Overall, 112 relevant publications were retrieved, which included 109 peer reviewed journal articles, 2 World Health Organization (WHO) technical reports, and 1 edited text book. A range of bacterial (Bartonella spp, Mycoplasma spp., Mycobacterium avium complex), viral (Dengue, SARS-CoV-2, Parvovirus-B19, Cytomegalovirus, Epstein-Barr virus), parasitic (Plasmodium spp., Babesia spp.), and fungal (Histoplasma spp.) infections were associated with IAH in SCD. There are two broad types of IAH in patients with SCD; infection associated extra-medullary haemolysis (IAEMH) and infection associated intra-medullary haemolysis (IAIMH). While IAEMH is associated with severe anaemia due to intravascular haemolysis caused by red cell invasion, oxidative injury, auto-antibodies, and/or pathogen-haem interaction, IAIMH is associated with haemophagocytic tri-lineage destruction of haematopoietic precursors in the bone marrow. Conclusion: Various microbial pathogens have been associated with IAH in SCD. SCD patients with fever, severe anaemia or pancytopenia should be investigated for early diagnosis and prompt treatment of IAH, which is a lifethreatening haematological emergency for which transfusion therapy alone may not suffice. Prompt and sustainable termination of IAH may require therapeutic combination of transfusion, anti-microbial chemotherapy, and immune modulation therapy. SCD patients should also receive counselling on hygiene, barrier protection against vectors, routine chemoprophylaxis for locally endemic diseases, and immunization for vaccine-preventable infections as a long-term preventive strategy against IAH.

Effects of Postmortem Hemolysis and Ultrafiltration on Creatinine Detection Results / 法医学杂志

OBJECTIVES@#To investigate the interference of postmortem hemolysis on the detection of creatinine and whether ultrafiltration can reduce the interference.@*METHODS@#A total of 33 non-hemolyzed whole blood samples from the left heart were collected. Hemolyzed samples with 4 hemoglobin mass concentration gradients H1-H4 were artificially prepared. Ultrafiltration was performed on each hemolyzed sample. Creatinine concentrations in non-hemolyzed serum (baseline serum), hemolyzed samples and ultrafiltrate were detected. Bias (B), Pearson correlation and receiver operator characteristic (ROC) of baseline creatinine concentration between before and after ultrafiltration were analyzed.@*RESULTS@#As the hemoglobin mass concentration increased, B of the hemolyzed samples in the H1-H4 groups gradually increased, the |B| was 2.41(0.82, 8.25)-51.31(41.79, 188.25), reaching a maximum of 589.06%, and there was no statistically significant between the creatinine concentration and the baseline creatinine concentration (P=0.472 7, r=0.129 5). After ultrafiltration of hemolyzed samples, the interference of creatinine concentration in ultrafiltrate was significantly reduced, the |B| was 5.32(2.26, 9.22)-21.74(20.06, 25.58), reaching a maximum of 32.14%, and there was a positive correlation with baseline creatinine concentration (P<0.05, r=0.918 2). In the hemolyzed samples of H3 and H4 groups, there were 7 false-positive samples and 1 false-negative sample; in the ultrafiltrate samples, there were no false-positive sample and 1 false-negative sample. ROC analysis results showed the hemolyzed samples were lack of diagnostic value (P=0.117 5).@*CONCLUSIONS@#The postmortem hemolysis significantly interferes creatinine detection results of blood samples, ultrafiltration can reduce hemolysis-induced interference in postmortem creatinine detection.

The Correlation Analysis between the Titer of IgG Anti-A/B Erythrocyte Antibody In Vivo of the Neonate and the Severity of Hemolytic Disease of Newborn / 中国实验血液学杂志

OBJECTIVE@#To investigate the titer of IgG anti-A/B erythrocyte antibody in vivo of the neonate with hemolytic disease of newborn(HDN), and explore its clinical valua in evaluating the severity of HDN.@*METHODS@#300 neonates with HDN, 50 neonates with neonatal hyperbilirubinemiain and 50 healthy neonates were selected as research object and Microtubes Gel Test was used to detect the titer of IgG anti-A/B erythrocyte antibody in vivo. Their clinical data and their mothers' prenatal examination data were retrospectively analyzed. Three hemolysis tests (direct antiglobulin test, free antibody test and release test), irregular antibody screening, and the titer of IgG anti-A/B blood group antibody was determined by serological method. Red blood cells(RBC), hemoglobin(Hb), reticulocytes(Ret) and nucleated red cells were detected by hematology analyzer. Indirect bilirubin and albumin(Alb) were detected by biochemical analyzer. The relationship between the titer of IgG anti-A/B erythrocyte antibody in vivo and the severity of HDN was analyzed.@*RESULTS@#There were six serological diagnosis modes in the HDN group,the difference between modes was statistically significant (P<0.05). The antibody titer relationship between HDN neonates and pregnant women was positive correlation(r=0.8302). The highest antibody titer of release test and free antibody test were 1∶32 and 1∶2, and the difference was statistically significant（P<0.05）. RBC, Hb and Alb in HDN patients were lower than those in neonatal hyperbilirubinemia patients and healthy neonates (P<0.05), and were negatively relevant with antibody titer in vivo (r=-0.8016). Bilirubin content in HDN patients were higher than those in neonatal hyperbiliru binemia patients and healthy neonates group(P<0.05), and was positively relevant with antibody titer in vivo (r=0.8731). The hospital day in HDN patients was significantly relevant with the antibody titer in vivo (r=0.8547), but not with the age, sex, weight and ABO blood types (P>0.05).@*CONCLUSION@#The detection of antibody titer in HDN patients can be used to evaluate the antibody concentration in vivo, predict the ability of antibody to induce erythrocyte hemolysis, and help to judge the serenrity and prognosis of HDN.

Effect of Anti-Oxidative of Ethyl Pyruvate and Taurine on the Red Blood Cell Storage at 4 ℃ / 中国实验血液学杂志

OBJECTIVE@#To investigate the anti-oxidative effect of ethyl pyruvate (EP) and taurine (TAU) on the quality of red blood cells stored at 4±2 ℃, hemolysis, energy metabolism and lipid peroxidation of the red blood cells in the preservation solution were studied at different intervals.@*METHODS@#At 4±2 ℃, the deleukocyte red blood cells were stored in the citrate-phosphate-dextrosesaline-adenine-1 (CPDA-1) preservation (control group), preservation solution with EP (EP-AS), and TAU (TAU-AS) for long-term preservation. The enzyme-linked immunoassay and automatic blood cell analyzer were used to detect hemolysis and erythrocyte parameters. Adenine nucleoside triphosphate (ATP), glycerol 2,3-diphosphate (2,3-DPG) and malondialdehyde (MDA) kits were used to test the ATP, 2,3-DPG and MDA concentration.@*RESULTS@#During the preservation, the rate of red blood cell hemolysis in EP-AS and TAU-AS groups were significantly lower than that in CPDA-1 group (P<0.01). The MCV of EP-AS group was increased with the preservation time (r=0.71), while the MCV of the TAU-AS group was significantly lower than that in the other two groups (P<0.05). The concentration of ATP and MDA in EP-AS and TAU-AS groups were significantly higher than that in CPDA-1 group at the 14th day (P<0.01). The concentrations of 2,3-DPG in the EP-AS and TAU-AS groups were significantly higher than that in the CPDA-1 group from the 7th day (P<0.01).@*CONCLUSION@#EP and TAU can significantly reduce the red blood cell hemolysis rate, inhibit the lipid peroxidation level of red blood cells, and improve the energy metabolism of red blood cells during storage. The mechanism of EP and TAU may be related to their antioxidation and membrane protection effect, so as to improve the red blood cell quality and extend the preservation time.

Análise qualitativa do teste de fragilidade osmótica para amostras processadas a fresco ou após 24 horas de incubação a 37ºC / Qualitative analysis of the osmotic fragility test for samples processed fresh or after 24 hours of incubation at 37ºC

Objetivo: Analisar qualitativamente o teste de fragilidade osmótica (F.O.) para amostras a fresco ou após incubação a 37°C. Métodos: Foram processadas 20 amostras de sangue periférico, coletadas em duplicata com 5mL em cada tubo com heparina, de pacientes com solicitação de F.O. como exame de rotina para processamento a fresco e após incubação por 24 horas em banho-maria a 37°C, em 13 tubos com concentrações variáveis de 0,1% a 0,9% de NaCl. Resultados: Foram analisadas 20 amostras de pacientes em sua maioria do gênero feminino 17/20 (85%), com idades entre 3 meses a 75 anos, para realização do teste de F.O. A análise qualitativa dos resultados mostrou que 9/20 (45%) amostras tiveram resultado concordante entre os testes de F.O. para amostras a fresco e após incubação a 37°C. Dos resultados discordantes, 8/11 (72,7%) resultados mostram fragilidade dos eritrócitos à hemólise nas amostras a fresco e curva normal (sem hemólise) após incubação da amostra a 37°C. Outros 3/11 (22,3%) resultados apresentaram curva normal (sem hemólise) no teste com amostra à fresco e resistência à hemólise no teste com a amostra após incubação a 37°C. Com o teste de Extato de Fisher não mostrou diferença estatística (p=0,5743) para as amostras processadas a fresco ou após incubação a 37°C. Conclusão: O teste de F.O. se mostrou mais eficiente quando a amostra testada foi analisada após incubação por 24 horas a 37°C em banho-maria, contudo não houve diferença estatística para resultados processados a fresco ou após incubação a 37°C.

Objective: Qualitatively analyze the osmotic fragility test (O.F.) for samples fresh or after incubation at 37°C. Methods: Twenty peripheral blood samples were processed, collected in duplicate with 5 ml in each tube with heparin, from patients with O.F. request. as a routine examination for fresh processing and after incubation for 24 hours in a water bath at 37°C, in 13 tubes with varying concentrations of 0.1% to 0.9% NaCl. Results: Twenty samples of patients were analyzed, mostly female, 17/20 (85%), aged between 3 months to 75 years, for the O.F. test. The qualitative analysis of the results showed that only 9/20 (45%) samples had a consistent result between the F.O. tests for fresh samples and after incubation at 37°C. From the discordant results, 8/11 (72.7%) results show fragility of erythrocytes to hemolysis in fresh samples and normal curve (without hemolysis) after sample incubation at 37o C. Other 3/11 (22.3%) results showed normal curve (without hemolysis) in the test with fresh sample and resistance to hemolysis in the test with the sample after incubation at 37o C. With the Fisher Extact test showing no statistical difference (p=0.5743) for samples processed fresh or after incubation at 37°C. Conclusion: The O.F. proved to be more efficient when the tested sample was analyzed after incubation for 24h at 37°C in a water bath, however, there was no statistical difference for results processed fresh or after incubation at 37°C.

Cord blood bilirubin and prediction of neonatal hyperbilirubinemia and perinatal infection in newborns at risk of hemolysis

Abstract Objective To assess the accuracy of umbilical cord bilirubin values to predict jaundice in the first 48 h of life and neonatal infection. Method Newborn infants treated at a regional well-baby nursery born at ≥36 weeks of gestation were included in this retrospective cohort study. All infants born in a 3-year period from mothers with O blood type and/or Rh-negative were included and had the umbilical cord bilirubin levels measured. Hyperbilirubinemia in the first 48 h was defined as bilirubin levels above the phototherapy threshold. Neonatal infection was defined as any antibiotic treatment before discharge. Results A total of 1360 newborn infants were included. Two hundred and three (14.9%) newborn infants developed hyperbilirubinemia in the first 48 h of life. Hyperbilirubinemic infants had smaller birth weight, higher levels of umbilical cord bilirubin, a higher rate of infection and were more often direct antiglobulin test positive. Umbilical cord bilirubin had a sensitivity of 76.85% and a specificity of 69.58% in detecting hyperbilirubinemia in the first 48 h, with the cut-off value at 34 µmol/L. The area under the receiver operating characteristic curve was 0.80 (95% CI: 0.78-0.82). Umbilical cord bilirubin had a sensitivity of 27.03% and specificity of 91.31% in detecting perinatal infection. The area under the receiver operating characteristic (ROC) curve was 0.59 (95% CI: 0.57-0.63). Conclusions A positive correlation was found between umbilical cord bilirubin and hyperbilirubinemia in the first 48 h of life. Umbilical cord bilirubin is a poor marker for predicting neonatal infection.

Prevalence and effect of intramammary infection due to coagulase-negative staphylococcal (CNS) species in somatic cell counts in milk from Holstein dairy cows in Boyaca, Colombia / Prevalencia y efecto de la infección intramamaria debida a especies de estafilococos coagulasa negativo (ECN) en el conteo de células somáticas en leche de vacas Holstein en Boyacá, Colombia

ABSTRACT Mastitis is one of the most important illnesses in specialized dairy herds worldwide due to the effects on production and animal health. The types caused by CNS has a special importance in a production where the main pathogens are controlled. The objective of the present work is to determine the prevalence of CNS in a dairy herd in Boyaca and also quantify the effects of every species of CNS in SCC. 40 cows were selected and sampled during 6 months, CMT was performed, and results from 1 to trace were sampled. The routine bacteriological test was also performed for CNS identification, and the isolating of CNS was performed through rpoB gene identification and through the type of strain using the pulse gel electrophoresis procedure. Out of 960 samples, 619 were positive for CNS growth. The most prevalent species were Staphylococcus epidermidis, S. chromogenes, S. sciuri, S. simulaans, S. haemolyticus and S. capitis. The results that were found here are similar to the results observed in different parts of the world, which confirms that they are pathogens that must be constantly evaluated because they can go unnoticed in routine controls, especially in those farms where major pathogens are not a serious problem. The results determined in this study demonstrate that CNS generates a slight increase in somatic cells.

RESUMEN La mastitis es una de las enfermedades más importantes en los rebaños lecheros especializados alrededor de todo el mundo debido a los efectos sobre la producción y la salud animal. Los tipos ocasionados por estafilococos coagualasa negativo (ECN) tienen una importancia especial en una producción en la que los principales patógenos están controlados. El objetivo del presente trabajo es determinar la prevalência del ECN en un hato lechero en Boyacá y cuantificar los efectos de cada especie de ECN en el conteo de células somáticas (CCS). Se seleccionaron 40 vacas y se tomaron muestras durante 6 meses, se realizó california mastitis test (CMT) y se tomaron muestras de los resultados desde 1 hasta donde hubo trazas. También se realizó la prueba bacteriológica de rutina para la identificación del ECN y el aislamiento del ECN se realizó mediante la identificación del gen rpoB y del tipo de cepa, usando el procedimiento de electroforesis en gel de pulso. De 960 muestras, 619 fueron positivas para el crecimiento del ECN. Las especies más prevalentes fueron Staphylococcus epidermidis, S. chromogenes, S. sciuri, S. simulans, S. haemolyticus y S. capitis. Los resultados encontrados aquí son similares a resultados en diferentes partes del mundo, lo que confirma que son patógenos que deben ser evaluados constantemente porque pueden pasar desapercibidos en los controles de rutina, especialmente en aquellas fincas donde los patógenos mayores no son un problema grave. Los resultados determinados en este estudio demuestran que el SNC genera un ligero aumento de células somáticas.

Evaluación de la sangre ovina como suplemento nutritivo en medios de cultivo: citratada y desfibrinada / Evaluation of sheep blood as nutritive suplement in culture media: citratated and defibrinated

Introducción: La sangre ovina constituye un suplemento esencial en la elaboración de medios de cultivo, dado que aporta factores nutricionales indispensables para el crecimiento y la recuperación de diversos microorganismos. Objetivo: Evaluar comparativamente el efecto de la sangre ovina, tanto citratada como desfibrinada, en medios de cultivo de base agar en cuanto al crecimiento bacteriano y la producción de hemólisis de cepas de referencia de diferentes bacterias patógenas, así como la recuperación o el aislamiento de microorganismos de muestras clínicas. Métodos: Se realizó un estudio observacional, descriptivo y transversal en 6 laboratorios de microbiología de la ciudad de Santiago de Cuba durante el año 2017, en los cuales se empleó sangre de ovinos de la raza pelibuey, para la elaboración de medios de cultivo. A cada laboratorio se le suministró tanto sangre citratada como desfibrinada y se le entregó una encuesta para valorar los resultados. Resultados: Existió un mejor crecimiento y aislamiento bacteriano en el medio suplementado con sangre desfibrinada, a pesar de que el rendimiento o los resultados en el caso de la sangre citratada resultaron satisfactorios. Conclusiones: Se confirmó la pertinencia del uso de la sangre desfibrinada como suplemento de enriquecimiento nutritivo en medios de cultivo; no obstante, quedó demostrada la utilidad de la sangre citratada en la labor de rutina del laboratorio de microbiología clínica.

Introduction: Sheep blood is an essential supplement in the elaboration of culture media, as it provides important nutritional factors for the growth and recovery of different organisms. Objective: To evaluate comparatively the effect of sheep citrated and defibrinated sheep blood in culture media with agar base as for the bacterial growth and the production of hemolysis in reference strains from different pathogen bacteria, as well as the recovery or isolation of microorganisms from clinical samples. Method: An observational, descriptive and cross-sectional was carried out in 6 microbiology laboratories in Santiago de Cuba city during 2017, in which male sheeps blood from the pelibuey breed for elaborating culture media. Each laboratory received either citrated blood or defibrinated and a survey was delivered to evaluate the results. Results: There was a better growing and bacterial isolation in the media supplemented with defibrinated blood, although yielding or results were favorable with citrated blood. Conclusions: The pertinence of the use of defibrinated blood as a supplement of nutritive enrichment in culture media was confirmed; however, the use of citrated blood was demonstrated in the routine work of the clinical microbiology laboratory.

Hemolytic anemia as the initial presentation of Wilson's disease. Regarding a rare disease with an infrequent presentation / Anemia hemolítica como presentación inicial de la enfermedad de Wilson. A propósito de una enfermedad rara con una presentación infrecuente

Abstract Wilson's disease is a rare genetic disorder that affects the excretion capacity of copper. Its distribution is worldwide, with an estimated prevalence in 30 cases per million habitants. Although the most frequent symptoms are those of hepatic and neuropsychiatric origin, hemolytic anemia with negative Coombs may be the only manifestation of the disease and its presentation usually precedes for months to clinically evident liver disease or neurological manifestations. The case of a young patient with negative Coombs hemolytic anemia and an alkaline phosphatase / total bilirubin ratio <4 and AST / ALT> 2.2 is presented, establishing Wilson's disease as a diagnosis. (Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1459).

Resumen La enfermedad de Wilson es un raro trastorno genético que afecta la capacidad de excreción del cobre. Su distribución es mundial, con una prevalencia estimada en 30 casos por millón de habitantes. Aunque los síntomas más frecuentes son los de origen hepático y neuropsiquiatricos, la anemia hemolítica con Coombs negativo puede ser la única manifestación de la enfermedad y su presentación suele preceder por meses a la enfermedad hepática clínicamente evidente o las manifestaciones neurológicas. Se presenta el caso de una paciente joven con anemia hemolítica Coombs negativo y relación fosfatasa alcalina/bilirrubina total <4 y AST/ALT >2,2, en quien terminó por establecerse como diagnóstico una enfermedad de Wilson.(Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1459).

Proposta de escala colorimétrica para inspeção visual do grau de hemólise em bolsas de concentrado de hemácias / Colorimetric scale proposal for visual inspection of the hemolysis degree in red cells concentrate bags

Objetivo: Os concentrados de hemácias não estão livres de lesões de estoque, sendo a mais comum a lise das hemácias. Criar uma escala colorimétrica para inspeção visual do grau de hemólise de concentrados de hemácias. Métodos: Foram utilizadas 102 bolsas de concentrados de hemácias (n=36), concentrados de hemácias pobres em leucócitos (n=36) ou de concentrados de hemácias desleucotizadas (n=36), com até 28 dias da produção, em condições padrões de armazenamento e da rotina da Fundação Hemopa. Os valores de hemoglobina e hematócrito foram lidos em contador automatizado. O grau de hemólise foi realizado em amostras de 5 mL do segmento das bolsas, com leitura em espectrofotômetro. O teste de hemólise foi realizado com três gotas de amostras do segmento das bolsas, em tubo seco, adição de 4 mL de solução fisiológica, centrifugação e inspeção visual do sobrenadante (estudo duplo-cego). Para a confecção da escala colorimétrica foram utilizados os resultados de grau de hemólise e teste de hemólise (n=36) e registro fotográfico dos vários níveis de grau de hemólise das bolsas. Para a validação da escala colorimétrica foram realizados grau de hemólise e teste de hemólise para as bolsas restantes (n=66), e com os resultados comparados com a escala proposta (estudo duplo-cego). Resultados: Apenas 7,5% (5/66) das bolsas testadas foram negativas para o teste de hemólise clássico (visual), e positivas quando reavaliadas na escala colorimétrica proposta. Conclusão: A escala colorimétrica proposta mostrou-se simples e reproduzível para definição de teste de hemólise e o grau de hemólise para concentrados de hemácias.

Objective: Red blood cell (RBC) concentrates are not free of stock lesions, the most common of which is lysis of RBC. Create a colorimetric scale for visual inspection of the degree of hemolysis of bag red blood cells. Methods: Eighty-two bags of RBC concentrates (n = 36), RBC concentrates poor in leukocytes (n = 36) or RBC concentrates desleucotizadas (n = 36) were used with up to 28 days of production, under standard storage conditions of the Hemopa Foundation routine. Hemoglobin and hematocrit values were read in automated counter. The degree of hemolysis performed in 5ml samples from the bag segment, with spectrophotometer reading. The hemolysis test was performed with three drops of bag segment samples, in a dry tube, addition of 4 mL of physiological solution, centrifugation and visual inspection of the supernatant (double-blind study). To prepare the colorimetric scale used was the degree of hemolysis and hemolysis test results (n = 36) and photographic records of multiple levels of degree of hemolysis of bags. For the validation of the Colorimetric scale, hemolysis degree and hemolysis test were performed for the remaining bags (n = 66), and the results were compared with the proposed scale (double-blind study). Results: Only 7.5% (5/66) of the tested bags were negative for the classical (visual) hemolysis test, and positive when reevaluated on the proposed colorimetric scale. Conclusion: The proposed colorimetric scale was simple and reproducible for the definition of hemolysis test and the degree of hemolysis for RBC concentrates.

Relationship between antifungal susceptibility profile and virulence factors in Candida albicans isolated from nail specimens

Abstract INTRODUCTION: The aim of this study was to evaluate some virulence factors in Candida albicans isolates from patients with onychomycosis and determine the correlation between these factors and the antifungal resistance profile. METHODS: Seventy species of C. albicans were confirmed using polymerase chain reaction amplification of the HWP1 gene. According to the Clinical & Laboratory Standards Institute guidelines, the susceptibility profile of four antifungal agents was investigated, and the production of aspartyl protease, phospholipase, haemolysin, and biofilm was determined. The correlation between these profiles was also investigated. RESULTS: The isolates indicated different levels of resistance and production of virulence factors. Significant correlations were observed between the minimum inhibitory concentration (MIC) of fluconazole/itraconazole and biofilm production, between phospholipase production and fluconazole/itraconazole MIC, and between fluconazole MIC and hemolytic activity in C. albicans isolates. The results also showed significant correlations between phospholipase activity and biofilm production. CONCLUSIONS: Our findings will contribute to a better understanding of the pathogenesis of C. albicans and characterize the relationship between virulence factors and antifungal resistance, which may suggest new therapeutic strategies considering the possible involvement of the virulence mechanism in the effectiveness of treatment.

Sudden death caused by Clostridium perfringens sepsis presenting as massive intravascular hemolysis

An 80-year-old Japanese woman with diabetes mellitus was admitted with gastrointestinal symptoms and pyrexia. At presentation, liver abscesses and severe hemolytic anemia were noted. Before detailed diagnostic evaluation and adequate treatment, she suddenly died 2.5 hours after admission. The autopsy and bacteriological examinations revealed liver abscesses and massive intravascular hemolysis caused by Clostridium perfringens as well as other miscellaneous critical pathological findings, including acute renal tubular necrosis, lung edema, and pulmonary fat embolism. In this article, the detailed autopsy results are described and clinicopathologic characteristics on Clostridium perfringens-related sudden death are discussed with a review of the literature.