Genetic and biochemical characterization of BIM-1, a novel acquired subgroup B1 MBL found in a Pseudomonas sp. strain from the Brazilian Amazon region.

OBJECTIVES: To characterize a novel acquired MBL, BIM-1, in a Pseudomonas #2 (subgroup P. guariconensis) strain isolated from the Aurá river located in the Brazilian Amazon hydrographic basin. METHODS: WGS using an Illumina® MiSeq System was used to characterize the genome of Pseudomonas sp. IEC33019 strain. Southern blotting/hybridization assays were performed to confirm the location of the MBL-encoding gene, blaBIM-1 (Belém Imipenemase). Antimicrobial susceptibility testing, cloning, and biochemical and phenotypic characterization were performed to determine BIM-1 kinetics. RESULTS: The IEC33019 strain showed high resistance rates to ß-lactams, ciprofloxacin and aminoglycosides, being susceptible only to polymyxins and susceptible, increased exposure to aztreonam. WGS analysis revealed a novel acquired MBL-encoding gene, blaBIM-1, found as a gene cassette inserted into a class 1 integron (In1326) that also carried qnrVC1 and aadA11e. In1326 was located in a complex transposon, Tn7122, carried by a 52.7 kb conjugative plasmid (pIEC33019) with a toxin/antitoxin system (vapB/vapC). BIM-1 belongs to the molecular subgroup B1 and shares 70.2% and 64.9% similarity with SIM-1 and IMP-1, respectively. Kinetics analysis of BIM-1 showed hydrolytic activity against all ß-lactams tested. CONCLUSIONS: BIM-1 is a novel acquired MBL encoded by a gene carried by mobile genetic elements, which can be transferred to other Gram-negative bacilli (GNB). Because the IEC33019 strain was recovered from a river impacted by a populous metropolitan region with poor basic sanitation and served by limited potable freshwater, it would be important to establish the role of the BIM-1-producing GNB as nosocomial pathogens and/or as colonizers of the riverside population in this geographical region.

Prevalencia y factores asociados a disfunción familiar en pacientes atendidos en el primer nivel de atención

Introducción: La funcionalidad familiar influye en el proceso salud-enfermedad, es por ello que se destaca su importancia en el primer nivel de atención. No existe evidencia concluyente sobre los factores que influyen en la disfunción familiar en pacientes atendidos en el primer nivel de atención. Objetivo: Determinar la prevalencia y factores asociados a disfunción familiar en pacientes atendidos en el primer nivel de atención. Métodos: Estudio transversal de análisis secundario de datos en pacientes atendidos en 7 establecimientos del primer nivel de atención de Lima, Perú, en 2019. Se utilizó el cuestionario Apgar Familiar y se indagó su asociación con factores demográficos-socioeconómicos. Se estimaron razones de prevalencia (RP) a través de modelos de regresión simple y múltiple. Resultados: De 150 pacientes, la mayoría fueron mujeres (81,3 %) y la mediana de edad fue de 32 años. El 14 % presentó disfunción familiar. Los pacientes con acceso a servicio de agua tenían menor prevalencia de disfunción familiar (RP: 0,04; IC95 %: 0,001 - 0,47). Residir entre 1 a 10 años en Lima representó menor prevalencia de disfunción familiar; en comparación con pacientes recién llegados a la capital (menos de 1 año) (RP: 0,15; IC95 %: 0,04 - 0,62). Conclusiones: La prevalencia de disfunción familiar en pacientes atendidos en primer nivel de atención es baja. Tener acceso a servicio de agua y residir entre 1 a 10 años en la capital influyó en una menor prevalencia de disfunción familiar.

Introduction: Family functionality influences the health-disease process, which is why its importance in the first level of care is highlighted. There is no conclusive evidence on the factors that influence family dysfunction in patients treated at the First Level of Care. Objective: To determine the prevalence and factors associated with family dysfunction in patients treated at the First Level of Care. Methods: Cross-sectional study of secondary data analysis in patients treated in 7 primary care establishments in Lima, Peru in 2019. The Family Apgar questionnaire was used, and its association with demographic-socioeconomic factors was investigated. Prevalence ratios (PR) were estimated through simple and multiple regression models. Results: Of 150 patients, the majority were women (81.3%) and the median age was 32 years. 14% presented family dysfunction. Patients with access to water service had a lower prevalence of family dysfunction (PR: 0.04; 95% CI: 0.001-0.47). Living between 1 and 10 years in Lima represented a lower prevalence of family dysfunction; compared with patients recently arrived in the capital (less than 1 year) (PR: 0.15; 95% CI: 0.04- 0.62). Conclusions: The prevalence of family dysfunction in patients treated at the primary care level was low. Having access to water service and residing between 1 and 10 years in the capital influenced a lower prevalence of family dysfunction.

Recommendations for Early and Comprehensive Management of Type 2 Diabetes and Its Related Cardio-Renal Complications.

Type 2 diabetes (T2D) is a global health problem accompanied by an elevated risk of complications, the most common being cardiac and renal diseases. In Lebanon, the prevalence of T2D is estimated at 8-13%. Local medical practice generally suffers from clinical inertia, with gaps in the yearly assessment of clinical manifestations and suboptimal screening for major complications. The joint statement presented here, endorsed by five Lebanese scientific medical societies, aims at providing physicians in Lebanon with a tool for early, effective, and comprehensive care of patients with T2D. Findings from major randomized clinical trials of antidiabetic medications with cardio-renal benefits are presented, together with recommendations from international medical societies. Optimal care should be multidisciplinary and should include a multifactorial risk assessment, lifestyle modifications, and a regular evaluation of risks, including the risks for cardiovascular (CV) and renal complications. With international guidelines supporting a shift in T2D management from glucose-lowering agents to disease-modifying drugs, the present statement recommends treatment initiation with metformin, followed by the addition of sodium-glucose cotransporter 2 inhibitors or glucagon-like peptide-1 receptor agonists due to their CV and renal protection properties, whenever possible. In addition to the selection of the most appropriate pharmacological therapy, efforts should be made to provide continuous education to patients about their disease, with the aim to achieve a patient-centered approach and to foster self-management and adherence to the medical plan. Increasing the level of patient engagement is expected to be associated with favorable health outcomes. Finally, this statement recommends setting an achievable individualized management plan and conducting regular follow-ups to monitor the patients' glycemic status and assess their risks every 3-6 months.

Toward New Epidemiological Landscapes of Trypanosoma cruzi (Kinetoplastida, Trypanosomatidae) Transmission under Future Human-Modified Land Cover and Climatic Change in Mexico.

Chagas disease, caused by the protozoa Trypanosoma cruzi, is an important yet neglected disease that represents a severe public health problem in the Americas. Although the alteration of natural habitats and climate change can favor the establishment of new transmission cycles for T. cruzi, the compound effect of human-modified landscapes and current climate change on the transmission dynamics of T. cruzi has until now received little attention. A better understanding of the relationship between these factors and T. cruzi presence is an important step towards finding ways to mitigate the future impact of this disease on human communities. Here, we assess how wild and domestic cycles of T. cruzi transmission are related to human-modified landscapes and climate conditions (LUCC-CC). Using a Bayesian datamining framework, we measured the correlations among the presence of T. cruzi transmission cycles (sylvatic, rural, and urban) and historical land use, land cover, and climate for the period 1985 to 2012. We then estimated the potential range changes of T. cruzi transmission cycles under future land-use and -cover change and climate change scenarios for 2050 and 2070 time-horizons, with respect to "green" (RCP 2.6), "business-as-usual" (RCP 4.5), and "worst-case" (RCP 8.5) scenarios, and four general circulation models. Our results show how sylvatic and domestic transmission cycles could have historically interacted through the potential exchange of wild triatomines (insect vectors of T. cruzi) and mammals carrying T. cruzi, due to the proximity of human settlements (urban and rural) to natural habitats. However, T. cruzi transmission cycles in recent times (i.e., 2011) have undergone a domiciliation process where several triatomines have colonized and adapted to human dwellings and domestic species (e.g., dogs and cats) that can be the main blood sources for these triatomines. Accordingly, Chagas disease could become an emerging health problem in urban areas. Projecting potential future range shifts of T. cruzi transmission cycles under LUCC-CC scenarios we found for RCP 2.6 no expansion of favourable conditions for the presence of T. cruzi transmission cycles. However, for RCP 4.5 and 8.5, a significant range expansion of T. cruzi could be expected. We conclude that if sustainable goals are reached by appropriate changes in socio-economic and development policies we can expect no increase in suitable habitats for T. cruzi transmission cycles.

Klebsiella Discitis in a 15-Year-Old Male Diagnosed With Plasma Microbial Cell-Free DNA Next-Generation Sequencing Test: A Case Report.

Spinal discitis (SD) is a rare condition, particularly in the pediatric population. The course of SD may be acute or chronic, and the non-specificity of symptoms leads to great delays in diagnosis. The most commonly isolated causative organism is Staphylococcus aureus whereas gram-negative infections are hardly ever reported in the literature. Comorbidities that increase the risk of bacteremia such as diabetes, chronic kidney disease, HIV, and cancer are major risk factors for SD. Hereby, we present an atypical case of SD in a previously healthy 15-year-old male with an unusual organism, Klebsiella aerogenes, diagnosed by plasma microbial cell-free DNA with negative blood cultures. The clinical course was complicated by antibiotic resistance and subsequent development of a ventral epidural abscess requiring readmission followed by surgical drainage of the abscess with a prolonged course of antibiotics.

Thermosonicated whey protein concentrate blends on quality attributes of reduced fat Panela cheese.

Aiming at producing a reduced fat cheese (RFC) as an alternative to full-fat Panela cheese, a highly consumed fresh Mexican dairy product, thermosonication (TS) processes (24 kHz, 400 W nominal power, 2, 4 and 6 min; 50, 55 and 60 °C) were evaluated to treat WPC (80% protein) blended with reduced-fat milk (1 and 2% fat), which were later LTLT pasteurized. TS blends were compared in terms of their technological properties (water holding capacity-WPC, gel firmness- GF, color, pH and titratable acidity) with those of a regular full fat (3%) LTLT pasteurized milk used as a control. Afterwards, a regression analysis was carried out with the obtained data in order to select the most appropriate conditions for cheesemaking purposes (similar GF, higher WHC with respect to the control), minimize both fat content and TS treatment duration to minimize energy expenses. According to these restrictions, the selected conditions were 1.5% fat milk-WPC blend, TS treated at 60 °C for 120 s; 1% fat milk-WPC blend, TS treated at 50 °C for 120 s and 1% fat milk-WPC blend, 50 °C for 144 s, which allowed preparing low fat cheeses (LFCs). These TS treatments were applied in a larger scale to elaborate Panela-type LFCs comparing different technological properties (cheese yield, syneresis, water content, texture profile analysis, color and titratable acidity) with those of a full fat variety, at day 1 and during 14 days of refrigerated storage. Results showed similar texture profiles of LFC cheeses and full fat milk cheeses throughout their storage period with significant changes in composition parameters (higher moisture, protein and salt contents, with low fat percentages), syneresis, selected color parameters (hue, b*), with no observed changes in cheese yield, TA and pH during cheese storage. These promising results are encouraging to develop LFCs with no physicochemical or technological defects using novel processing techniques that may help reducing calorie consumption without compromising sensory acceptability.

Biomonitoring of the hematological, biochemical and genotoxic effects of the mixture cypermethrin plus chlorpyrifos applications in bovines.

External antiparasitic agents applied in bovine production represent a risk to consumers of meat products, especially if the conditions of their use are not strictly respected. The post-mortem control of residues in meat is an activity that must be updated and reinforced by the biomonitoring of live animals and the use of analytical tools to help identifying signs of early warning risks. The objective of the present study was to carry out a pre-slaughter biomonitoring approach in Aberdeen Angus cattle and crosses (n = 12) with the application of a commercial formulation of cypermethrin plus chlorpyrifos. This was performed with a single therapeutic dose applied on the backs of the cattle, through hematological, enzymatic, as well as hepatic and renal function analysis in plasma, and then quantifying the genotoxic effect on lymphocytes. Analytical measurements of plasma concentrations of cypermethrin plus chlorpyrifos at 24 h were negative and therefore a low absorption of the compounds was assumed. Measurement of acetyl cholinesterase showed no inhibition after exposure. The concentration of urea increased between 24 h and 168 h post application of the formulation, without showing any kidney damage. The rest of the parameters analyzed did not show any variations. This evaluation of hematological and biochemical effects and of cytokinesis-block micronucleus cytome assay in bovines is proposed as a pre-slaughter control of biomonitoring of the health status of animals, with a focus on food safety for meat consumers.

Symptomatology and quality of life between two populations of climacteric women.

PURPOSE: The aim of this study is to compare climacteric symptomatology and sociodemographic conditions and their effect on quality of life in two populations: Monterrey (Mexico) and Madrid (Spain). METHODS: 469 women from Monterrey (mean age 50.5 + 4.3 years) and 452 (mean age 51.7 + 3.7 years) from Madrid participated in the study. Descriptive analyses of sociodemographic and clinics characteristics of the sample were performed. A cross-sectional design and a regression analysis were performed to establish the sociodemographic and clinical variables that would be used as predictors of quality of life. Data was collected using the Menopause-Specific Quality of Life, MENQOL, the Menopause Rating Scale (MRS), the Hospital Anxiety and Depression Scale (HADS), the Quality of Life Scale for Women Aged From 45 to 64 (QLS), and a sociodemographic and clinical interview designed ad hoc. RESULTS: Approximately 60% of both Spanish and Mexican women present symptoms during climacteric that impairs their quality of life. Spanish women suffer more intense symptoms and for a longer period of time than Mexican women, with the exception of anxiety. Mexican women report better quality of life than Spanish women and it is moderated by educational, socioeconomical, and marital status. Women's knowledge about menopause is also related to a better quality of life. CONCLUSIONS: Our study confirms the differences in climacteric symptomatology between populations and the impact of educational level and knowledge about menopause as predictors of a better quality of life in climacteric women.

Evaluation of the serum procalcitonin level as an indicator of severity and mortality in abdominal sepsis due to secondary peritonitis.

BACKGROUND: Procalcitonin is a biomarker of sepsis, whose concentrations increase when some endotoxin enters the bloodstream. It is used, among other things, to discriminate the etiology of infections, increase or decrease the antibiotic spectrum, and predict mortality. OBJECTIVE: To determine the utility of the serum level of procalcitonin as a predictor of severity and mortality. METHOD: Retrospective, descriptive, cross-sectional study of patients diagnosed with abdominal sepsis during the period from April 2016 to February 2017. In all cases the severity was determined by APACHE II, SOFA, Mannheim and CONUT, and mortality. The sample was divided into those with procalcitonin > 10.1 and < 10. RESULTS: We included 99 cases (41 female and 58 male). The main organ causing abdominal sepsis was the appendix 56%. The mean of procalcitonin for the sample was 7.94 (standard deviation: ± 13.76). The findings, subjected to statistical verification by means of the Mann-Whitney U test, showed statistical significance among the cases with procalcitonin 10.1, with the Mannheim scores > 26 points (p = 0.003), CONUT > 6 points (p = 0.027) and presence of organic faults (p = 0.001), but not with APACHE, SOFA and mortality. CONCLUSIONS: Procalcitonin is related to the severity determined by the Mannheim index, CONUT and the development of organic faults.

INTRODUCCIÓN: La procalcitonina es un biomarcador de sepsis, cuyas concentraciones aumentan cuando alguna endotoxina entra en el torrente sanguíneo. Se emplea, entre otras cosas, para discriminar la etiología de las infecciones, escalar/desescalar antibióticos, aumentar o disminuir el espectro antibiotico, y predecir la mortalidad. OBJETIVO: Determinar si los valores de procalcitonina se relacionan con la gravedad y la mortalidad de los pacientes con sepsis abdominal. MÉTODO: Estudio retrospectivo, descriptivo y transversal de pacientes con diagnóstico de sepsis abdominal, de abril de 2016 a febrero de 2017. En todos los casos se determinó la gravedad mediante las escalas APACHE II, SOFA, Mannheim y CONUT, así como la mortalidad. Se dividió la muestra en pacientes con procalcitonina > 10.1 y < 10. RESULTADOS: Se incluyeron 99 casos (41 mujeres y 58 hombres). El principal órgano causante de sepsis abdominal fue el apéndice (56%). La media de procalcitonina para la muestra se situó en 7.94 (desviación estándar: ± 13.76). Los hallazgos, sometidos a verificación estadística mediante la prueba U de Mann-Whitney, mostraron una relación estadísticamente significativa en los casos con procalcitonina 10.1, puntajes de Mannheim > 26 puntos (p = 0.003), CONUT > 6 puntos (p = 0.027) y presencia de fallas orgánicas (p = 0.001); no hubo relación significativa entre los valores de procalcitonina y los puntajes de las escalas APACHE II y SOFA, ni con la mortalidad. CONCLUSIÓN: La gravedad medida por la procalcitonina se relaciona con la gravedad determinada mediante los índices de Mannheim y CONUT, y con el desarrollo de fallas orgánicas.

Reference values for acetyl and butyrylcholinesterases in cattle under actual management conditions, hepatic and renal function by application of chlorpyrifos.

Chlorpyrifos is an anticholinesterase organophosphate insecticide widely used in Argentina in the production of food derived from animal, fruit and horticultural origin and is reported as a residue within these products. Local reference values for acetyl and butyrylcholinesterase were determined in Aberdeen Angus bovine and cross bred cattle (n = 25), a requirement to be able to evaluate toxicity of commercial organophosphate and carbamate formulations. The activity of cholinesterase enzymes presented an overall mean of 2,183.00 ± 485.6 IU L-1 for erythrocyte acetylcholinesterase and 203.1 ± 42.06 IU L-1 for plasma butyrylcholinesterase, which are used as reference values for meat steers within a system of intensive production in a semi-arid region. The toxic potential of chlorpyrifos in steers of the same breeds (n = 12) was assessed applying chlorpyrifos 15.00% Tipertox® in a single therapeutic dose of 7.50 mg kg-1 by topical route. Prior to application and then on day 1 and day 21 post-application, both blood cholinesterases, serum chlorpyrifos concentration by ultra-high resolution liquid chromatography with mass detector, analysis of blood counts, total proteins, liver enzymes, urea and creatinine were evaluated. The mean plasma concentration of chlorpyrifos was 27.90 ug L-1 at 24 h. The findings indicate that the therapeutic treatment of castrated male bovines treated with chlorpyrifos, applied by pour-on according to the manufacturer's instructions, does not cause changes in the variables evaluated.

Evaluation of a microfluidic flow assay to screen for von Willebrand disease and low von Willebrand factor levels.

Essentials von Willebrand factor (VWF) function is shear stress dependent. Platelet accumulation in a microfluidic assay correlates with VWF levels. The microfluidic assay discriminates type 1 von Willebrand disease from healthy controls. The microfluidic flow assay detects responses to therapeutic intervention (DDAVP). SUMMARY: Background von Willebrand disease (VWD) is a mucocutaneous bleeding disorder with a reported prevalence of 1 in 10 000. von Willebrand factor (VWF) function and platelet adhesion are regulated by hemodynamic forces that are not integrated into most current clinical assays. Objective We evaluated whether a custom microfluidic flow assay (MFA) can screen for deficiencies in VWF in patients presenting with mucocutaneous bleeding. Methods Whole blood from individuals with mucocutaneous bleeding was assayed in a custom MFA. Results Thirty-two patients with type 1 VWD (10/32) or reported mucocutaneous bleeding were enrolled. The platelet adhesion velocity (r = 0.5978 for 750 s-1 and 0.6895 for 1500 s-1 ) and the maximum platelet surface area coverage (r = 0.5719 for 750 s-1 and 0.6633 for 1500 s-1 ) in the MFA correlated with VWF levels. Furthermore, the platelet adhesion velocity at 750 s-1 (type 1 VWD, mean 0.0009761, 95% confidence interval [CI] 0.0003404-0.001612; control, mean 0.003587, 95% CI 0.002455-0.004719) and at 1500 s-1 (type 1 VWD, mean 0.0003585, 95% CI 0.00003914-0.0006778; control, mean 0.003132, 95% CI 0.001565-0.004699) differentiated type 1 VWD from controls. Maximum platelet surface area coverage at 750 s-1 (type 1 VWD, mean 0.1831, 95% CI 0.03816-0.3281; control, mean 0.6755, 95% CI 0.471-0.88) and at 1500 s-1 (type 1 VWD, mean 0.07873, 95% CI 0.01689-0.1406; control, mean 0.6432, 95% CI 0.3607-0.9257) also differentiated type 1 VWD from controls. We also observed an improvement in platelet accumulation after 1-desamino-8-d-arginine vasopressin (DDAVP) treatment at 1500 s-1 (pre-DDAVP, mean 0.4784, 95% CI 0.1777-0.7791; post-DDAVP, mean 0.8444, 95% CI 0.7162-0.9726). Conclusions These data suggest that this approach can be used as a screening tool for VWD.

The small-molecule MERTK inhibitor UNC2025 decreases platelet activation and prevents thrombosis.

Essentials Signaling by Gas6 through Tyro3/Axl/Mer receptors is essential for stable platelet aggregation. UNC2025 is a small molecule inhibitor of the Mer tyrosine kinase. UNC2025 decreases platelet activation in vitro and thrombus formation in vivo. UNC2025's anti-platelet effect is synergistic with inhibition of the ADP receptor, P2Y12 . SUMMARY: Background Growth arrest-specific protein 6 signals through the TAM (TYRO-3-AXL-MERTK) receptor family, mediating platelet activation and thrombus formation via activation of the aggregate-stabilizing αIIb ß3 integrin. Objective To describe the antithrombotic effects mediated by UNC2025, a small-molecule MERTK tyrosine kinase inhibitor. Methods MERTK phosphorylation and downstream signaling were assessed by immunoblotting. Light transmission aggregometry, flow cytometry and microfluidic analysis were used to evaluate the impact of MERTK inhibition on platelet activation and stability of aggregates in vitro. The effects of MERTK inhibition on arterial and venous thrombosis, platelet accumulation at microvascular injury sites and tail bleeding times were determined with murine models. The effects of combined treatment with ADP-P2Y1&12 pathway antagonists and UNC2025 were also evaluated. Results and Conclusions Treatment with UNC2025 inhibited MERTK phosphorylation and downstream activation of AKT and SRC, decreased platelet activation, and protected animals from pulmonary embolism and arterial thrombosis without increasing bleeding times. The antiplatelet effect of UNC2025 was enhanced in combination with ADP-P2Y1&12 pathway antagonists, and a greater than additive effect was observed when these two agents with different mechanisms of inhibition were coadministered. TAM kinase signaling represents a potential therapeutic target, as inhibition of this axis, especially in combination with ADP-P2Y pathway antagonism, mediates decreased platelet activation, aggregate stability, and thrombus formation, with less hemorrhagic potential than current treatment strategies. The data presented here also demonstrate antithrombotic activity mediated by UNC2025, a novel translational agent, and support the development of TAM kinase inhibitors for clinical applications.

A microfluidic model of hemostasis sensitive to platelet function and coagulation.

Hemostasis is the process of sealing a vascular injury with a thrombus to arrest bleeding. The type of thrombus that forms depends on the nature of the injury and hemodynamics. There are many models of intravascular thrombus formation whereby blood is exposed to prothrombotic molecules on a solid substrate. However, there are few models of extravascular thrombus formation whereby blood escapes into the extravascular space through a hole in the vessel wall. Here, we describe a microfluidic model of hemostasis that includes vascular, vessel wall, and extravascular compartments. Type I collagen and tissue factor, which support platelet adhesion and initiate coagulation, respectively, were adsorbed to the wall of the injury channel and act synergistically to yield a stable thrombus that stops blood loss into the extravascular compartment in ~7.5 min. Inhibiting factor VIII to mimic hemophilia A results in an unstable thrombus that was unable to close the injury. Treatment with a P2Y12 antagonist to reduce platelet activation prolonged the closure time two-fold compared to controls. Taken together, these data demonstrate a hemostatic model that is sensitive to both coagulation and platelet function and can be used to study coagulopathies and platelet dysfunction that result in excessive blood loss.

Comparison of foam swabs and toothbrushes as oral hygiene interventions in mechanically ventilated patients: a randomised split mouth study.

INTRODUCTION: During critical illness, dental plaque may serve as a reservoir of respiratory pathogens. This study compared the effectiveness of toothbrushing with a small-headed toothbrush or a foam-headed swab in mechanically ventilated patients. METHODS: This was a randomised, assessor-blinded, split-mouth trial, performed at a single critical care unit. Adult, orally intubated patients with >20 teeth, where >24 hours of mechanical ventilation was expected were included. Teeth were cleaned 12-hourly using a foam swab or toothbrush (each randomly assigned to one side of the mouth). Cleaning efficacy was based on plaque scores, gingival index and microbial plaque counts. RESULTS: High initial plaque (mean=2.1 (SD 0.45)) and gingival (mean=2.0 (SD 0.54)) scores were recorded for 21 patients. A significant reduction compared with initial plaque index occurred using both toothbrushes (mean change=-1.26, 95% CI -1.57 to -0.95; p<0.001) and foam swabs (mean change=-1.28, 95% CI -1.54 to -1.01; p<0.001). There was significant reduction in gingival index over time using toothbrushes (mean change=-0.92; 95% CI -1.19 to -0.64; p<0.001) and foam swabs (mean change=-0.85; 95% CI -1.10 to -0.61; p<0.001). Differences between cleaning methods were not statistically significant (p=0.12 for change in gingival index; p=0.24 for change in plaque index). There was no significant change in bacterial dental plaque counts between toothbrushing (mean change 3.7×104 colony-forming units (CFUs); minimum to maximum (-2.5×1010 CFUs, 8.7×107 CFUs)) and foam swabs (mean change 9×104 CFUs; minimum to maximum (-3.1×1010 CFUs, 3.0×107 CFUs)). CONCLUSIONS: Patients admitted to adult intensive care had poor oral health, which improved after brushing with a toothbrush or foam swab. Both interventions were equally effective at removing plaque and reducing gingival inflammation. TRIAL REGISTRATION NUMBER: NCT01154257; Pre-results.

Microangiopathic hemolytic anemia due to ADAMTS-13 loss in idiopathic systemic capillary leak syndrome.

Essentials Idiopathic systemic capillary leak syndrome (SCLS) is characterized by episodes of vascular leakage. We present the case of a patient with SCLS who developed microangiopathic hemolytic anemia (MAHA). We propose that this anemia is the result of ADAMTS-13 loss in the third-space fluid. This suggests that MAHA can occur in patients with significant extravasation of proteins. SUMMARY: Idiopathic systemic capillary leak syndrome (SCLS) is a rare process characterized by acute and recurrent episodes of vascular leakage with severe hypotension, hypoalbuminemia, hemoconcentration and edema. Anemia and thrombocytopenia are not part of this syndrome, but here we present the case of a pediatric patient with a clinical presentation consistent with SCLS who subsequently developed microangiopathic hemolytic anemia at a time when she had significant fluid loss and anasarca. Based on serial ADAMTS-13 levels, we propose that the anemia in this patient developed as a result of ADAMTS-13 loss in the third-space fluid, a novel mechanism for acquired microangiopathic hemolytic anemia.

Analysis of the Enzymatic Activity of an NS3 Helicase Genotype 3a Variant Sequence Obtained from a Relapse Patient.

The hepatitis C virus (HCV) is a species of diverse genotypes that infect over 170 million people worldwide, causing chronic inflammation, cirrhosis and hepatocellular carcinoma. HCV genotype 3a is common in Brazil, and it is associated with a relatively poor response to current direct-acting antiviral therapies. The HCV NS3 protein cleaves part of the HCV polyprotein, and cellular antiviral proteins. It is therefore the target of several HCV drugs. In addition to its protease activity, NS3 is also an RNA helicase. Previously, HCV present in a relapse patient was found to harbor a mutation known to be lethal to HCV genotype 1b. The point mutation encodes the amino acid substitution W501R in the helicase RNA binding site. To examine how the W501R substitution affects NS3 helicase activity in a genotype 3a background, wild type and W501R genotype 3a NS3 alleles were sub-cloned, expressed in E. coli, and the recombinant proteins were purified and characterized. The impact of the W501R allele on genotype 2a and 3a subgenomic replicons was also analyzed. Assays monitoring helicase-catalyzed DNA and RNA unwinding revealed that the catalytic efficiency of wild type genotype 3a NS3 helicase was more than 600 times greater than the W501R protein. Other assays revealed that the W501R protein bound DNA less than 2 times weaker than wild type, and both proteins hydrolyzed ATP at similar rates. In Huh7.5 cells, both genotype 2a and 3a subgenomic HCV replicons harboring the W501R allele showed a severe defect in replication. Since the W501R allele is carried as a minor variant, its replication would therefore need to be attributed to the trans-complementation by other wild type quasispecies.