Alpha-thalassemia is a hereditary hemoglobin disorder characterized by reduced or absent α-globin gene, and its severity is associated with the number of affected alleles. Several methods are available for detecting α-thalassemia, such as multiplex ligation-dependent probe amplification (MLPA) and PCR-based hybridization strip assay. Multiplex PCR offers a faster, more convenient, and cost-effective alternative. In this study, we aimed to optimize a current PCR-based method for α-thalassemia screening and evaluate its utility using clinical samples. We also investigated the prevalence and spectrum of common mutations responsible for α-thalassemia in Thailand and Korea. A total of 1261 samples from Thailand, 560 samples from different ethnic groups residing in Korea, and 300 samples from native Koreans were collected and tested. The concordance rate between the data collected in Thailand and in this study was 99.92%. Further, approximately 5.9% of the non-Korean individuals living in Korea were identified as healthy carriers, whereas no mutations were observed in Koreans. Comparing the data with MLPA or Sanger sequencing data showed 100% agreement rate in both cases. We successfully developed a PCR method for the diagnosis of α-thalassemia that is fast, less labor-intensive, and cost-effective. Given the performance results of this method, it has great potential for application in α-thalassemia diagnosis.
Purpose: This study was designed to investigate the relationship between peripheral blood inflammatory markers and prognosis in MDS patients.Methods: We conducted a study involving 183 MDS patients who were diagnosed at Taizhou Hospital of Zhejiang Province and Enze Hospital between January 2015 and December 2019. The end point of follow-up was September 2022. To minimize the impact of other confounding factors among the 110 included MDS patients, X-tile software was used to determine the optimal cutoff points for peripheral blood inflammation markers. Based on these cutoff points, the cohort of patients was divided into a high-risk group and a low-risk group. The OS in each group was analyzed by the Kaplan–Meier method, and univariate and multivariate Cox regression analyses were employed.Results: The MDS patients included 73 men and 37 women with a median age of 72 years (32–92 years). The median OS was 28 months (1–83 months), 17 patients (15.45%) experienced conversion to AML, and 94 patients (85.45%) died during the follow-up period. The optimal cutoff points were ALC (1.2 × 109/L), AMC (0 × 109/L), CRP (6.1 mg/L), MLR (0.125), NLR (2.25) and PLR (71.4). Patients in the ALC (≤1.2 × 109/L, P = 0.017), MLR (>0.125, P = 0.01), PLR (>71.43, P = 0.044), and CRP (>6.1 mg/L, P < 0.0001) groups had shorter overall survival. The MLR (>0.125, P = 0.011) and CRP (>6.1 mg/L, P = 0.017) levels were related to poor prognosis.Conclusions: Elevated MLR and CRP levels may be independent indicators of poor prognosis in newly diagnosed MDS patients.
Mark A. Sammut, Mohammed E. F. Rahman, Claire Bridge
et al.
Dual antithrombotic therapy (DAT) without aspirin reduces bleeding compared with triple antithrombotic therapy (TAT) in patients with atrial fibrillation who have undergone percutaneous coronary intervention, without apparently increasing ischemic events. A prospective pharmacodynamic study was performed to investigate the impact of aspirin on bleeding time, platelet function and fibrin clot analysis in this population. Patients receiving TAT (n = 16), comprising aspirin, ticagrelor/prasugrel and a direct-acting oral anticoagulant (DOAC), were compared with those receiving DAT without aspirin (n = 18). Bleeding time was reduced with DAT compared with TAT (median 27.8 vs 30.0 minutes, p = .005). Assessed by light transmission aggregometry, median platelet aggregation was significantly increased with DAT compared with TAT in response to arachidonic acid (63 vs 3%, p = .002) and collagen (72 vs 37%, p < .001) but not 5-μmol/L adenosine diphosphate (25 vs 27%, p = .966) or thrombin-receptor-activating peptide (37 vs 24%, p = .086). VerifyNow P2Y12 assay showed > 70% inhibition in all patients. Fibrin clot lysis time and maximum turbidity were similar between groups. Using P2Y12 inhibitors of consistent potency, DAT improves hemostasis through sparing cyclooxygenase-1-mediated platelet activation but has a comparable effect to TAT on other pathways and fibrin clot properties. DAT with ticagrelor/prasugrel and DOAC may provide sufficient antithrombotic effect without excessive anti-hemostatic effect.
Abstract Background Bone marrow (BM) evaluation is the de facto standard for diagnosis, molecular analysis, risk stratification, and therapy response assessment in acute myeloid leukemia (AML), but in patients with a high number of circulating blast cells, the peripheral blood (PB) sample could provide similar information as BM. However, there is no large‐scale molecular study comparing the two specimens in terms of their gene expression profiles, cellular heterogeneities, and ex‐vivo drug sensitivity. Methodology We used (i) the BEAT‐AML cohort each with detailed molecular data; (ii) cell‐type deconvolution to estimate leukemic and immune cell proportions between specimen types; (iii) differential expression (DE) and drug‐cell type association analysis; and (iv) logistic regression models to assess the association between induction therapy response, cell‐type composition and first‐line drug treatment. Results Results: We identified 207 patients having BM and 116 patients having PB samples. There was a total of 1271 DE genes (false discovery rate < 0.05) between BM and PB; the top enriched pathways in terms of DE genes belong to the immune system pathways. Aggregated ex‐vivo drug response profiles from the two specimens were largely similar, as were the cellular components, except for the GMP‐like cell type (17% in BM vs. 5% in PB, p‐value = 2 × 10−7). Among the specimen‐specific results, the GMP‐like subtype was associated with multiple drug resistance in BM and the ProMono‐like subtype in PB. Several cell types were associated with the response to induction therapy, but the impact of specimen type on the interaction of cell type and cytarabine‐associated induction therapy was not statistically significant for most cell types. Results Conclusions: Even though there are molecular and cellular differences between BM and PB samples, they show many similarities in ex‐vivo drug response profiles, indicating the clinical utility of the substantially less‐invasive PB samples.
Objetivos: Hemofilia A e B são caracterizadas como a deficiência dos fatores de coagulação VIII e IX, respectivamente. O objetivo do presente trabalho é realizar uma revisão da literatura sobre a artropatia hemofílica. Material e métodos: Revisão de literatura através das bases de dados Google Scholar e Pubmed em agosto de 2021. Foram utilizadas as seguintes palavras-chave: hemophilia, hemophilia AND arthropathy, hemophilia AND hematology. Resultados: Hemorragia intra-articular espontânea é a manifestação mais frequente na hemofilia grave (pacientes com níveis de fator de coagulação menor que 1 IU/dL). Recorrências na mesma articulação causam sinovite, gerando artropatia crônica, com perda de movimento articular em 80% dos pacientes. A ocorrência aumenta conforme a progressão da idade, sendo 33% e 47% em crianças com hemofilia A e B, respectivamente, enquanto ocorre em 60% e 42% dos adultos com hemofilia A e B, respectivamente. Os locais mais acometidos são tornozelo, cotovelo e joelho. A sinovite ocorre devido aos produtos da degradação do sangue na articulação ultrapassarem a capacidade da membrana sinovial de absorvê-las e eliminá-las, principalmente o ferro e hemossiderina, os quais induzem a produção de citocinas inflamatórias, além de contribuir para a reabsorção e remodelamento do osso e da cartilagem. A sinovite hemofílica é caracterizada pela proliferação do tecido sinovial e de vasos sanguíneos. Essa afeta consideravelmente a qualidade de vida, prejudicando principalmente a mobilidade, a prática de esportes e o lazer. Discussão: A deficiência dos fatores VIII e IX causa redução e atraso da geração de trombina, levando a discrasia sanguínea que ocasiona sangramentos os quais afetam as articulações. Quando o fator de coagulação é menor que 1 IU/dL, o paciente apresenta hemofilia grave, a qual constitui cerca de 50% dos casos diagnosticados, sendo que o risco de hemorragias aumenta proporcionalmente ao aumento da deficiência do fator de coagulação. Nesse caso, os sangramentos articulares ocorrem frequentemente e deve-se reconhecer os sintomas da artropatia hemofílica que, inicialmente, são o edema da articulação e dor não traumática que desaparecem quando o episódio agudo acaba. A profilaxia da artropatia hemofílica consiste na terapia de reposição do fator coagulante. Quando a artropatia já está estabelecida, o tratamento inclui fisioterapia, uso de órteses, injeções intra-articulares e tratamento cirúrgico. Conclusão: As hemofilias, principalmente as graves, são fator precipitante ao desenvolvimento de artropatia nos pacientes portadores, logo, o diagnóstico precoce é essencial para possibilitar o manejo adequado e evitar o surgimento da artropatia. Ademais, o conhecimento da alta prevalência da artropatia em hemofílicos é fundamental para a consideração de profilaxia e proteção articular. Como observado, o acometimento articular crônico prejudica a qualidade de vida do paciente, impossibilitando a mobilidade adequada e realização de atividades físicas. Nas crianças, esse cenário é danoso ao desenvolvimento físico e social, sendo necessário estar atento às crianças portadoras de hemofilia que passem a apresentar algum distúrbio locomotor e rapidamente agir em prol da resolução da condição.
INTRODUCTION Secondary Acute Myeloid Leukemia (s-AML) evolves from a previous hematopoietic clonal disease such as Myelodysplastic Syndromes (MDS/AML), myeloproliferative neoplasia (NPM/AML), medullary insufficiencies - aplastic anemia - (AA/AML) or exposure to chemo or radiotherapy (t-AML). The objective of this work is to describe and highlight the demographic, pathophysiologic and clinical-therapeutic characteristics of s-AML patients compared with p-AML. METHODS This is a retrospective cohort study based on the casuistry from 34 reference centers in Latin America during a period of 10 years (JAN10'-MAY19'). Patients ≥18 years old with primary AML, excluding the promyelocytic subtype (p-AML), and s-AML were admitted. Age, gender, performance status, comorbidity, cytogenetics, mutations, AML subtypes, extramedullary compromise, treatments and overall survival (OS) were analyzed. Statistically, Graph Pad Prism version 5.00 and, SPSS version 17 were used. RESULTS One thousand eleven patients with newly diagnosed AML were recruited, 693 (68.5%) corresponded to p-AML and 318 (31.5%) to s-AML. The demographic differences between p-AML and s-AML are shown on Table 1. Subtypes of s-AML: t-AML (18.5%), MDS/AML (58.2%), NMP/AML (13.5%), AA/AML (5.7%) and others s-AML (4.1%). Global median age was 58 years (R 18-93) and male 52%. Extramedullary compromise in CNS (3.2%) and other organs (5.5%). Seven hundred ninety-three cytogenetic studies were evaluable (based on MRC classification): High (22.3%), Intermediate (68.3%) and Low Risk (9.3%). FLT3 mutation was more frequently found in p-AML. In s-AML, the multivariate study showed short overall survival associated with ECOG ≥2 (HR:2.0), white blood count ≥ 50x109/L at diagnosis (HR:1.9), poor risk karyotype (HR:1.6) and age over 60 years (HR:1.5). At least, 883 patients received treatment (Table 2). During this study period, 211 patients (21%) were transplanted; 49 (23%) were s-AML; histoidentical related donor (46%), haploidentical (39%), non-related (8%) and autologous (7%). The median survival for all AML was 11.0 months with a statistically significant difference in favor of the p-AML (Figure 1). CONCLUSION Performance status (by ECOG ≥2), age ≥60, level of leukocytes a ≥50x109/L, poor risk karyotype and s-AML subtype at diagnosis had a significant worse impact on overall survival. Most patients with s-AML came from MDS, they were older and their incidence increased as the population aged. They presented more comorbidities and worse performance status. Undoubtedly, our findings showed that s-AML is a distinct high risk subset of myeloid disorder with adverse prognosis and represents a therapeutic challenge. No relevant conflicts of interest to declare.
Federica D’asta, Fenella Halstead, Paul Harrison
et al.
The infection of a wound is one of the major contributors to delays in healing and tissue regeneration. As multi-drug resistance to antibiotics is becoming a serious threat, research in this field has focused on finding new agents and strategies to fight infection and additionally to reduce healing times. The topical use of autologous Platelet Rich Plasma (PRP) as a biological accelerator of the healing process, has been safely used as a form of treatment for wounds since the 1990s. Although the presence or absence of leucocytes in PRP preparation was previously neglected, in the last decade more attention has been paid to their role and several studies have been conducted to explore both their immuno-metabolic effects and their antimicrobial properties. In this review, we aim to summarise the literature on the contribution of leucocytes included in PRP preparations in terms of their antimicrobial properties. This should help to inform clinical practice and additional research in this promising field.