A scalable insect cell-based production process of the human recombinant BMX for in-vitro covalent ligand high-throughput screening.

Bone Marrow Tyrosine kinase in the chromosome X (BMX) is a TEC family kinase associated with numerous pathological pathways in cancer cells. Covalent inhibition of BMX activity holds promise as a therapeutic approach against cancer. To screen for potent and selective covalent BMX inhibitors, large quantities of highly pure BMX are normally required which is challenging with the currently available production and purification processes. Here, we developed a scalable production process for the human recombinant BMX (hrBMX) using the insect cell-baculovirus expression vector system. Comparable expression levels were obtained in small-scale shake flasks (13 mL) and in stirred-tank bioreactors (STB, 5 L). A two-step chromatographic-based process was implemented, reducing purification times by 75% when compared to traditional processes, while maintaining hrBMX stability. The final production yield was 24 mg of purified hrBMX per litter of cell culture, with a purity of > 99%. Product quality was assessed and confirmed through a series of biochemical and biophysical assays, including circular dichroism and dynamic light scattering. Overall, the platform herein developed was capable of generating 100 mg purified hrBMX from 5 L STB in just 34 days, thus having the potential to assist in-vitro covalent ligand high-throughput screening for BMX activity inhibition

Lavandula luisieri and Lavandula viridis essential oils as upcoming anti-protozoal agents: A key focus on leishmaniasis

Background and objectives: Leishmania species is the causative agent of leishmaniasis, a broad-spectrum clinical condition that can even be life-threatening when neglected. Current therapeutic strategies, despite beings highly cost-effective, have been increasingly associated with the appearance of drug-resistant microorganisms. Thus, an increasing number of thorough studies are needed towards upcoming drug discovery. This study aims to reveal the anti-protozoa activity of Lavandula luisieri and Lavandula viridis essential oils (EO) and their main components (1,8-cineole, linalool, and borneol). Materials and Methods: L. luisieri and L. viridis EO and their main components' leishmanicidal effects were tested in vitro against Leishmania infantum, Leishmania major, and Leishmania tropica strains. Cell viability effects were estimated by using the tetrazolium-dye (MTT) colorimetric method, morphological changes were assessed by scanning electron microscopy (SEM) and ultrastructural investigation by transmission electronic microscopy (TEM). Phosphatidylserine externalization, mitochondrial membrane potential (MMP), and cathepsin D activity assessment were also carried out. Finally, cytotoxic activity of the studied matrices was also determined in mammalian cells. Results: Plant-studied EO exhibited prominent anti-Leishmania effects (IC50 = 31-263 µg/mL), with L. luisieri being the most active one. At concentrations corresponding to IC50 values, EO-exposed L. infantum promastigotes suffered marked ultrastructural modifications. The presence of aberrant-shaped cells, mitochondrial and kinetoplast swelling, and autophagosomal structures were the most common evidenced changes. L. luisieri EO exerted its leishmanicidal activity through different mechanisms, but mainly through unleashing apoptosis. Phosphatidylserine externalization, mitochondrial membrane potential loss, and cell-cycle arrest at G(0)/G(1) phase were the most remarkable apoptosis-mediated aspects. Inhibition of cathepsin D activity was also observed. No toxic effects were found on macrophage cells. Conclusions: L. luisieri seems to be an upcoming source of bioactive molecules for leishmaniasis control and to find leading molecules for new drugs formulation against Leishmania infections.This work was supported by the European Regional Development Fund (ERDF), through the Centro 2020 Regional Operational Programme under project CENTRO-01-0145-FEDER-000008:BrainHealth 2020, and through the COMPETE 2020—Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT—Fundação para a Ciência e a Tecnologia, I.P., under strategic project POCI-01-0145-FEDER-007440 (UID/NEU/04539/2013). Acknowledgments: The authors are grateful to Jorge Paiva for helping in plant taxonomy; to José Correia da Costa from Centro de Imunologia e Biologia Parasitária, Instituto Nacional Ricardo Jorge, Porto for supplying L. infantum (zymodeme MON-1); to António Osuna, Departamento de Parasitología, Facultad de Ciencias, Instituto de Biotecnología, Universidad de Granada, for supplying L. major. FCT for Strategic project ref. UID/BIM/04293/2013, and “NORTE2020—Programa Operacional Regional do Norte” (NORTE-01-0145-FEDER-000012)

Seroprevalence of Bordetella pertussis in healthcare workers in a Portuguese tertiary referral hospital

info:eu-repo/semantics/publishedVersio

Determinants of tuberculosis transmission and treatment abandonment in Fortaleza, Brazil

BACKGROUND: Tuberculosis (TB) remains a public health problem, despite recent achievements in reducing incidence and mortality rates. In Brazil, these achievements were above the worldwide average, but marked by large regional heterogeneities. In Fortaleza (5th largest city in Brazil), the tuberculosis cure rate has been declining and treatment abandonment has been increasing in the past decade, despite a reduction in incidence and an increase in directly observed therapy (DOT). These trends put efforts to eliminate tuberculosis at risk. We therefore sought to determine social and programmatic determinants of tuberculosis incidence and treatment abandonment in Fortaleza. METHODS: We analyzed sociodemographic and clinical data for all new tuberculosis cases notified in the Notifiable Diseases Information System (SINAN) from Fortaleza between 2007 and 2014. We calculated incidence rates for 117 neighborhoods in Fortaleza, assessed their spatial clustering, and used spatial regression models to quantify associations between neighborhood-level covariates and incidence rates. We used hierarchical logistic regression models to evaluate how individual- and neighborhood-level covariates predicted tuberculosis treatment abandonment. RESULTS: There were 12,338 new cases reported during the study period. Case rates across neighborhoods were significantly positively clustered in two low-income areas close to the city center. In an adjusted model, tuberculosis rates were significantly higher in neighborhoods with lower literacy, higher sewerage access and homicide rates, and a greater proportion of self-reported black residents. Treatment was abandoned in 1901 cases (15.4%), a rate that rose by 71% between 2007 and 2014. Abandonment was significantly associated with many individual sociodemographic and clinical factors. Notably, being recommended for DOT was protective for those who completed DOT, but associated with abandonment for those who did not. CONCLUSION: Low socioeconomic status areas have higher tuberculosis rates, and low socioeconomic individuals have higher risk of treatment abandonment, in Fortaleza. Treatment abandonment rates are growing despite the advent of universal DOT recommendations in Brazil. Proactive social policies, and active contact tracing to find missed cases, may help reduce the tuberculosis burden in this setting

Effects of Low-Moderate Load High-Velocity Resistance Training on Physical Performance of Under-20 Futsal Players

Resistance training (RT) is an effective methodology to improve physical performance of athletes. However, up to now, no studies have addressed the RT benefits in under-20 futsal players. The purpose of this study was to evaluate the effects of six weeks of RT with high-velocity movements, low-to-moderate loads, and low volume on physical performance of under-20 futsal players. A total of 21 players were divided into two groups: A control group (CG, n = 10) and a RT group (RTG, n = 11). The RTG performed two weekly training sessions constituted by leg-press, jumps, and sprints, along with three futsal training sessions, while the CG only performed the futsal training. Before and after the intervention, the sprint time in 0⁻10 m (T10), 10⁻20 m (T10⁻20), and 0⁻20 m (T20), the countermovement jump (CMJ) height, the T-Test time, the kicking ball speed (KBS), and the maximum dynamic strength in the leg-press, were assessed. In post-test, significant improvements in CMJ, T-Test, KBS, and leg-press were found for the RTG, whilst a significant decrease in T10⁻20 was evidenced in the CG. The present results suggested that RT based on high velocity movements, low-to-moderate loads, and low volume produce positive effects on physical performance of under-20 futsal players.info:eu-repo/semantics/publishedVersio

The effect of warm-up on sprinting kinematics

The purpose of this study was to verify the effects of warm-up on kinematic variables during short distance repeated sprints. Twenty-two college students randomly performed 2 x 30-m running time-trials after warm-up or with no warm-up, in different days. Performance (time-trial) and biomechanical (step length and step frequency) were assessed during both repeated trials. Performance was 0.5% faster after warm-up in the first 30-m time-trial (p = 0.03, d = 0.44), but without differences on step length and frequency. The second sprint was not different between conditions, but it was better than the first sprint in the no warm-up condition. This condition also led to higher changes between the first and second sprint. Thus, the warm-up is suggested to improve maximal running performances and maintaining kinematics more similar throughout the sprints.N/

Small bowel pseudomelanosis and oral iron therapy

Small bowel pseudomelanosis is a rarely reported clinical entity characterized by brown pigmentation of small bowel mucosa. The authors describe two cases, both with iron deficiency anemia, one of an 81-year-old female patient submitted for capsule endoscopy that revealed a brown pigmentation of all small bowel mucosa and another of an 81-year-old male whose retrograde double-balloon enteroscopy revealed a diffuse brown pattern of small bowel mucosa. Ileal biopsies confirmed intense iron deposition in the macrophages of the lamina propria. Both patients were on oral iron therapy and the second one had a previous double-balloon enteroscopy, 2 years earlier, which revealed only ileal angiodysplasias. These two cases demonstrate the importance of two new endoscopic methods for diagnosis of small bowel pseudomelanosis, the rarity of such an entity and its close relation with oral iron therapy

Office Hysteroscopy After Ultrasonographic Diagnosis of Thickened Endometrium in Postmenopausal Patients

The aim of our study was to access office hysteroscopy results in postmenopausal patients with thickened endometrium. A retrospective descriptive study was carried out on 245 postmenopausal patients submitted to office hysteroscopy after sonographic diagnosis of thickened endometriumin 20 consecutive months.Women were evaluated for age, hormonal therapy, hysteroscopic findings, procedure duration, complications and associated pain, and histological diagnosis. Patients with and without uterine bleeding were considered separately. Symptomatic patients were older and had longer procedure duration. The most frequent hysteroscopic finding was endometrial polyp in both groups. Pain was subjectively assessed in a numeric scale from 0 to 10 and median value was 4. There were no complications reported. Global neoplasia rate was 2.9% for asymptomatic patients and 16.4% for symptomatic ones (p<0.05). Thickened endometrium with postmenopausal metrorrhagia gave patients a significantly higher risk for neoplasia and hyperplasia

Centrality Dependence of Hadron Multiplicities in Nuclear Collisions in the Dual Parton Model

We show that, even in purely soft processes, the hadronic multiplicity in nucleus-nucleus interactions contains a term that scales with the number of binary collisions. In the absence of shadowing corrections, this term dominates at mid rapidities and high energies. Shadowing corrections are calculated as a function of impact parameter and the centrality dependence of mid-rapidity multiplicities is determined. The multiplicity per participant increases with centrality with a rate that increases between SPS and RHIC energies, in agreement with experiment.Comment: 15 pages, 4 postscript figure