IGHV gene mutational status and LPL/ADAM29 gene expression as clinical outcome predictors in CLL patients in remission following treatment with oral fludarabine plus cyclophosphamide

Several prognostic factors can predict the rapid progression in chronic lymphocytic leukaemia (CLL), including IGHV mutational status, cytogenetic abnormalities and, more recently, LPL/ADAM29 expression. In contrast, few studies have been devoted to the influence of these factors on clinical outcome in responding patients after therapy. We here propose to analyse the impact of IGHV gene status, LPL and ADAM29 gene expression on disease-free survival (DFS) and overall survival (OS) in 41 stage B or C CLL patients in remission after oral fludarabine plus cyclophosphamide. The median follow-up was of 64 (16–74) months. Sequencing of IGHV showed mutated (M) VH genes in 16 of 41 cases and unmutated (UM) in 25 cases. Analysis of LPL and ADAM29 expression in 35 of 41 cases showed overexpression of ADAM29 in 17 cases (14 M and three UM) and LPL in 18 cases (all UM). Patients expressing UM IGHV and LPL had shorter DFS and OS when compared to patients expressing M IGHV and/or ADAM29. Furthermore, blood minimal residual disease (MRD) evaluation using four-colour flow cytometry was performed in 33 out the 41 patients. We showed that patients who achieved phenotypic remission displayed longer DFS than those with MRD⁺. Our results support the use of LPL and ADAM29 gene expression associated to IGHV mutational status for predicting the clinical outcome of patients treated by oral fludarabine + cyclophosphamide and could be considered for treatment strategies.     

Case Reports on Emergency Treatment of Cardiovascular Syndromes Through Heparin‐Mediated Low‐Density Lipoprotein/Fibrinogen Precipitation: A New Approach to Augment Cerebral and Myocardial Salvage

Abstract: We report the first experiences with HELP apheresis as an emergency treatment for acute cardiovascular syndromes; two patients who were not eligible for lysis therapy and catheter intervention were treated with HELP apheresis instead. Both patients had a most severe, generalized atherosclerosis and reached the hospital too late for conventional measures. In both cases, the use of the apheresis dramatically improved the clinical situation to such an extent that the possibilities of this apheresis system urge further investigation.     

Early dysfunction and long-term improvement in endothelium-dependent vasodilation in the infarct-related artery after thrombolysis

Endothelial function is thought to be an important factor in the pathogenesis of atherosclerosis, hypertension and heart failure. In the 1990s, high-frequency ultrasonographic imaging of the brachial artery to assess endothelium-dependent flow-mediated vasodilation (FMD) was developed. The technique provokes the release of nitric oxide, resulting in vasodilation that can be quantitated as an index of vasomotor function. The noninvasive nature of the technique allows repeated measurements over time to study the effectiveness of various interventions that may affect vascular health. However, despite its widespread use, there are technical and interpretive limitations of this technique. State-of-the-art information is presented and insights are provided into the strengths and limitations of high-resolution ultrasonography of the brachial artery to evaluate vasomotor function, with guidelines for its research application in the study of endothelial physiology.     

A Highly Specific Holin-Mediated Mechanism Facilitates the Secretion of Lethal Toxin TcsL in Paeniclostridium sordellii

Protein secretion is generally mediated by a series of distinct pathways in bacteria. Recently, evidence of a novel bacterial secretion pathway involving a bacteriophage-related protein has emerged. TcdE, a holin-like protein encoded by toxigenic isolates of Clostridioides difficile, mediates the release of the large clostridial glucosylating toxins (LCGTs), TcdA and TcdB, and TpeL from C. perfringens uses another holin-like protein, TpeE, for its secretion; however, it is not yet known if TcdE or TpeE secretion is specific to these proteins. It is also unknown if other members of the LCGT-producing clostridia, including Paeniclostridium sordellii (previously Clostridium sordellii), use a similar toxin-release mechanism. Here, we confirm that each of the LCGT-producing clostridia encode functional holin-like proteins in close proximity to the toxin genes. To characterise the respective roles of these holin-like proteins in the release of the LCGTs, P. sordellii and its lethal toxin, TcsL, were used as a model. Construction and analysis of mutants of the P. sordellii tcsE (holin-like) gene demonstrated that TcsE plays a significant role in TcsL release. Proteomic analysis of the secretome from the tcsE mutant confirmed that TcsE is required for efficient TcsL secretion. Unexpectedly, comparative sample analysis showed that TcsL was the only protein significantly altered in its release, suggesting that this holin-like protein has specifically evolved to function in the release of this important virulence factor. This specificity has, to our knowledge, not been previously shown and suggests that this protein may function as part of a specific mechanism for the release of all LCGTs.     

Tracking Demographic Movements and Immunization Status to Improve Children’s Access to Immunization: Field-Based Randomized Controlled Trial

BackgroundCountries’ Expanded Program on Immunization (EPI) contribute to the reduction of mortality and morbidity, but access to these vaccines remains limited in most low-income countries.ObjectiveWe aim to assess whether involving community volunteers (CVs) to track children’s vaccination status and demographic movements and using recorded data to plan catch-up immunization sessions can improve children’s vaccination timeliness, completeness, and coverage.MethodsThis was a field-based randomized controlled trial and communities of the Foumban health district in West Cameroon were allocated to intervention or control groups. In the intervention group, a CV per community was trained to visit households monthly for a year to assess and record in a register, details of EPI-targeted children, their demographic movements and immunization status. The scanned recorded pages were sent to the health center immunization team through WhatsApp and used to organize monthly community catch-up immunization sessions. In the control group, EPI vaccination sessions were routinely conducted. Surveys were conducted at 6 and 12 months from the beginning of the intervention in both study groups to assess and compare immunization timeliness, coverage, and completeness.ResultsOverall, 30 buildings per cluster were surveyed at midline and endline. Of the 633 and 729 visited households in the intervention group at midline and endline, 630 (99.5%) and 718 (98.4%), respectively, consented to participate. In the control group, 507 and 651 households were visited and 505 (99.6%) and 636 (97.7%), respectively, consented to participate. At 12 months intervention, the month one timeliness of bacille Calmette–Guerin (BCG) vaccine did not increase in the intervention group compared with the control group for the age groups 0-11 months (adjusted odds ratio [aOR] 1.1, 95% CI 0.7-1.8) and 0-59 months (aOR 1.1, 95% CI 0.9-1.4), and significantly increased for the first-year BCG vaccine administration for the age group 0-23 months (aOR 1.5, 95% CI 1.1-2.2). The coverage of diphtheria-pertussis-tetanus and hepatitis B+Hemophilus influenzae type B (DPT-Hi +Hb) dose 3 (aOR 2.0, 95% CI 1.5-2.7) and of DPT-Hi+Hb dose 1 (aOR 1.8, 95% CI 1.4-2.4) vaccines increased significantly in the intervention group compared with the control group in the age groups 12-59 months and 12-23 months, respectively. Specific (DPT-Hi+Hb dose 1 to DPT-Hi+Hb dose 3: aOR 1.9, 95% CI 1.4-2.6) and general (BCG to measles: aOR 1.5, 95% CI 1.1-2.1) vaccine completeness increased significantly in the intervention group compared with the control group.ConclusionsFindings support that involving CVs to track children’s vaccination status and demographic movements and using recorded data to plan catch-up immunization sessions improve children’s vaccination timeliness, completeness, and coverage. This strategy should be adopted to improve access to vaccination for EPI target populations and the consistency verified in other contexts.Trial RegistrationPan African Clinical Trials Registry PACTR201808527428720; https://pactr.samrc.ac.za/TrialDisplay.aspx?TrialID=3548     

Risk of Seven-Day Worsening and Death: A New Clinically Derived COVID-19 Score

This monocentric, retrospective, two-stage observational study aimed to recognize the risk factors for a poor outcome in patients hospitalized with SARS-CoV-2 infection, and to develop and validate a risk score that identifies subjects at risk of worsening, death, or both. The data of patients with SARS-CoV-2 infection during the first wave of the pandemic were collected and analyzed as a derivation cohort. Variables with predictive properties were used to construct a prognostic score, which was tried out on a validation cohort enrolled during the second wave. The derivation cohort included 494 patients; the median age was 62 and the overall fatality rate was 22.3%. In a multivariable analysis, age, oxygen saturation, neutrophil-to-lymphocyte ratio, C-reactive protein and lactate dehydrogenase were independent predictors of death and composed the score. A cutoff value of 3 demonstrated a sensitivity (Se), specificity (Sp), positive predictive value (PPV) and negative predictive value (NPV) of 93.5%, 68.5%, 47.4% and 97.2% for death, and 84.9%, 84.5%, 79.6% and 87.9% for worsening, respectively. The validation cohort included 415 subjects. The score application showed a Se, Sp, PPV and NPV of 93.4%, 61.6%, 29.5% and 98.1% for death, and 81%, 76.3%, 72.1% and 84.1% for worsening, respectively. We propose a new clinical, easy and reliable score to predict the outcome in hospitalized SARS-CoV-2 patients.