Cultivos celulares primarios de Lutzomyia shannoni (Diptera: psychodidae) y estudio cariologico preliminar de la especie

To obtain a Lutzomyia shannoni (Dyar) cell line for viral susceptibility studies and parasite maintainance, primary cell cultures of this species were initiated. This species is a vector for vesicular stomatitis virus in the United States and a suspected vector of cutaneous leishmaniasis in the Americas. Starting with embryos and phlebotominae neonate larvae, embryonic tissue explants in a MMIVP12 medium were carried out. After being supplemented with 20% fetal bovine serum and an antibiotic and antimycotic mixture, they were incubated at 28°C in a non-CO, environment. Cell growth began 85 to 88 days after the cultures were initiated dueto epithelial cell vesicle presence; these were floating in the media or adhered to small fragments of tissue with divided cells. Following mechanical stimulation of the cultures, cell proliferation increased in the week following the procedure; however, cell mitotic process was slow, similar to that developed with Lu. longipalpis, but different to cell cultures derived from mosquitoes. The formation of individual colonies, dispersed on the surface of the culture flasks, was observed 90 days after incubation; these later evolved to a semiconfluent monolayer. The cellular morphology was heterogeneous with predominant epithelial types. Using the squash technique, this species' karyotype was obtained; the number of diploid chromosomes was 8, derived from fourth-instar larvae brain tissue.Con el propósito de obtener una línea celular de Lutzomyia shannoni (Dyar) para estudios de susceptibilidad viral y mantenimiento de parásitos, se iniciaron cultivos celulares primarios de esta especie, vectora del virus de la estomatitis vesicular en los Estados Unidos y vectora sospechosa de leishmaniasis cutánea en las Américas. A partir de embriones y larvas neonatas del flebotomineo, se realizaron explantes de tejidos embrionarios en el medio MMIVP12, suplementado con 20% de suero fetal bovino y una mezcla de antibiótico y antimicótico, los cuales fueron incubados a una temperatura promedio de 2VC, sin atmósfera de CO,. El crecimiento celular comenzó en un periodo de 85 a 88 días después de efectuadas las siembras, mediante la presencia de vesículas compuestas de células epitelioides, flotando en el medio o adheridas a pequeños fragmentos de tejidos con células en división. Previa estimulación mecánica de los cultivos, se incrementó la proliferación celular a la semana siguiente de efectuado el procedimiento; sin embargo, el proceso mitótico de las células fue lento, similar al desarrollado con Lu. longipalpis, pero diferente a los cultivos celulares derivados de mosquitos. La formación de colonias individuales, dispersas en la superficie del frasco de cultivo, se observó a los 90 días de incubación, las cuales posteriormente evolucionaron a una monocapa semiconfluente. La morfología celular fue heterogénea con predominio de tipos epitelioides. Mediante la técnica de squash, se obtuvo el cariotipo de la especie, cuyo número diploide de cromosomas fue de 8, derivados de tejidos cerebrales de larvas de IV estadio

Relação entre a proteína C ultrarreativa, diabetes e doença periodontal em pacientes com ou sem infarto do miocárdio

Artículo original362-368Objective: The purpose of this study was to evaluate the impact of diabetes and periodontal disease in us-CRP, an inflammatory marker in patients with and without acute myocardial infarction (AMI). Subjects and methods: A case-control study was conducted in 401 subjects aged between 30 and 75 years, living in Bogotá D.C. (Colombia). Patients arriving at the emergency room of the San Ignacio University Hospital with AMI were included into the case group. The control group was defined as those subjects without AMI. The following blood tests were performed: complete blood count (CBC), glycemia, total cholesterol, triglycerides, cHDL, cLDL, and us-CRP. Patients with infections or antibiotic treatment within the last three months, who had received periodontal treatment within the six months prior to the study entry, had oral ulcerations, or less than seven teeth were excluded from the study. Periodontal disease was diagnosed based on the 1999 Armitage’s classification. Results: The mean us-CRP value found in diabetic patients with severe chronic periodontitis was 5.31 mg/L (SD 6.82), and 2.38 mg/L (SD 4.42) in non-diabetic patients, being statistically significant (p = 0.000). Conclusion: Diabetes had an impact in periodontal disease and us-CRP. In patients with AMI, DM and PD considerably increased the us-CRP

Comparison of Values of C-Reactive Protein in Edentulous Patients and Patients with Chronic Moderate-to-Advanced Periodontal Disease at the Pontificia Universidad Javeriana Dental School in Bogotá

Antecedentes: La enfermedad periodontal (EP) es un factor de riesgo para desarrollar enfermedadescardiovasculares. Puede influir e iniciar una reacción autoinmune, aumentandola inflamación sistémica y acelerando la progresión de placas ateroescleróticas prexistentes.Ante inflamación aumenta la concentración de proteína C-reactiva (medida por PCR-us),que está relacionada con ateroesclerosis y riesgo cardiovascular. Se ha encontrado queel valor de PCR-us es significativamente mayor en pacientes con periodontitis. Objetivo:Determinar si existen diferencias significativas en los valores de PCR-us de pacientes conEP crónica entre moderada y avanzada no tratada y pacientes edéntulos totales comomarcador de ausencia de EP. Métodos: El diseño fue de casos y controles con una muestrade 60 pacientes mayores de 30 años de edad (30 casos con periodontitis crónica moderadaa avanzada y 30 controles edéntulos totales). Se tomó una muestra de sangre a todos lospacientes (cuadro hemático, colesterol, triglicéridos, glucemia, PCR-us) y se analizaron loshallazgos. Resultados: El promedio de PCR-us en los pacientes con periodontitis fue 2,19mg/L, y en los pacientes edéntulos, de 4,12 mg/L. Existe una tendencia a hallar valoresde PCR-us más elevados en pacientes edéntulos, al considerar que se encontró mayorexposición al riesgo en los pacientes con periodontitis. Los resultados no fueron estadísticamentesignificativos. Conclusión: La PCR-us se presentó más aumentada en los pacientesedéntulos totales y los valores de PCR-us en pacientes con periodontitis no se observaroncomo un factor de riesgo elevado para enfermedad cardiovascular. Background: Periodontal disease (PD) is a risk factor for cardiovascular disease. It caninitiate an autoimmune reaction, increase systemic inflammation, and accelerate the progressionof preexisting atherosclerotic plaques. In presence of inflammation, PD increasesthe concentration of C-reactive protein (measured through hs-CRP) that is associated withatherosclerosis and cardiovascular risk. It has been found that the value of hs-CRP is significantlyhigher in patients with periodontitis. Objective: Determine if there are significantdifferences in the values of hs-CRP among patients with untreated moderate-to-advancedchronic PD and edentulous patients (marker of absence of PE). Methods: A case-controlstudy was carried out with a sample of 60 patients older than 30 years of age (30 cases withmoderate-to-advanced chronic periodontitis diagnosed and 30 edentulous controls). Bloodsamples were taken from all patients (complete blood count, cholesterol, triglycerides, glucose,hs-CRP) and the results were compared. Results: The average hs-CRP in patients withperiodontitis was 2.19 mg/L and 4.12 mg/L in edentulous patients. There is a tendency tofind values higher of hs-CRP in edentulous patients, given that there was higher exposure inpatients with periodontitis. The results were not statistically significant. Conclusion: hs-CRPwas higher in edentulous patients and hs-CRP values in patients with periodontitis were notseen as a high risk factor for cardiovascular disease.95-10

Association between periodontal disease and endothelial dysfunction in smoking patients

Q329-35Over the past two decades, there has been increasing interest in the impact of oral health on cardiovascular disease, particularly regarding the effects of chronic infections such as periodontitis on the endothelium. The aim of this study was to evaluate in healthy smokers whether there are any significant differences in the frequency of endothelial dysfunction between subjects with chronic moderate to severe periodontal disease and periodontally healthy subjects. An observational cross-sectional study was conducted. The target population was adults older than 40 years of age. Blood tests were performed to determine values of CBC, glycaemia, total cholesterol, HDL-C, and LDLC. Periodontal examinations and probing were conducted with a Florida Probe®, and standardized procedures were used to measure flow-mediated dilation. Out of 150 subjects [69 male (46%) and 81 female (54%)], 75 (50%) had chronic periodontitis. The mean value for baseline flow-mediated dilation was 4.04% and the mean value for final flow-mediated dilation was 4.66%, with a 0.62% mean difference showing a statistically significant increase (p<0.001).This study found no significant difference in the flow-mediated dilation values between periodontally healthy subjects and those with periodontitis, in contrast to the literature, which suggests a negative impact of periodontal disease on endothelial function

Perfusion-weighted software written in Python for DSC-MRI analysis

IntroductionDynamic susceptibility-weighted contrast-enhanced (DSC) perfusion studies in magnetic resonance imaging (MRI) provide valuable data for studying vascular cerebral pathophysiology in different rodent models of brain diseases (stroke, tumor grading, and neurodegenerative models). The extraction of these hemodynamic parameters via DSC-MRI is based on tracer kinetic modeling, which can be solved using deconvolution-based methods, among others. Most of the post-processing software used in preclinical studies is home-built and custom-designed. Its use being, in most cases, limited to the institution responsible for the development. In this study, we designed a tool that performs the hemodynamic quantification process quickly and in a reliable way for research purposes.MethodsThe DSC-MRI quantification tool, developed as a Python project, performs the basic mathematical steps to generate the parametric maps: cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT), signal recovery (SR), and percentage signal recovery (PSR). For the validation process, a data set composed of MRI rat brain scans was evaluated: i) healthy animals, ii) temporal blood–brain barrier (BBB) dysfunction, iii) cerebral chronic hypoperfusion (CCH), iv) ischemic stroke, and v) glioblastoma multiforme (GBM) models. The resulting perfusion parameters were then compared with data retrieved from the literature.ResultsA total of 30 animals were evaluated with our DSC-MRI quantification tool. In all the models, the hemodynamic parameters reported from the literature are reproduced and they are in the same range as our results. The Bland–Altman plot used to describe the agreement between our perfusion quantitative analyses and literature data regarding healthy rats, stroke, and GBM models, determined that the agreement for CBV and MTT is higher than for CBF.ConclusionAn open-source, Python-based DSC post-processing software package that performs key quantitative perfusion parameters has been developed. Regarding the different animal models used, the results obtained are consistent and in good agreement with the physiological patterns and values reported in the literature. Our development has been built in a modular framework to allow code customization or the addition of alternative algorithms not yet implemented

Sources of Pre-Analytical Variations in Yield of DNA Extracted from Blood Samples: Analysis of 50,000 DNA Samples in EPIC

The European Prospective Investigation into Cancer and nutrition (EPIC) is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS) and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88%) performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies

Non-alcoholic beverages and risk of bladder cancer in Uruguay

BACKGROUND: Bladder cancer is the fourth most frequent malignancy among Uruguayan men. A previous study from Uruguay suggested a high risk of bladder cancer associated with maté drinking. We conducted an additional case-control study in order to further explore the role of non-alcoholic beverages in bladder carcinogenesis. METHODS: In the time period 1996–2000, 255 incident cases with transitional cell carcinoma of the bladder and 501 patients treated in the same hospitals and in the same time period were frequency matched on age, sex, and residence. Both cases and controls were face-to-face interviewed on occupation, tobacco smoking, alcohol drinking and intake of maté, coffee, tea, and soft drinks. Statistical analysis was carried out by unconditional multiple logistic regression. RESULTS: Ever maté drinking was positively associated with bladder cancer (odds ratio [OR] 2.2, 95% confidence interval [CI] 1.2–3.9) and the risk increased for increasing duration and amount of maté drinking. Both coffee and tea were strongly associated with bladder cancer risk (OR for coffee drinking 1.6, 95% CI 1.2–2.3; OR for tea drinking 2.3, 95% CI 1.5–3.4). These results were confirmed in a separate analysis of never-smokers. CONCLUSION: Our results suggest that drinking of maté, coffee and tea may be risk factors for bladder carcinoma in Uruguay

Reducing the global burden of cerebral venous thrombosis:An international research agenda

Background:Due to the rarity of cerebral venous thrombosis (CVT), performing high-quality scientific research in this field is challenging. Providing answers to unresolved research questions will improve prevention, diagnosis, and treatment, and ultimately translate to a better outcome of patients with CVT. We present an international research agenda, in which the most important research questions in the field of CVT are prioritized.Aims:This research agenda has three distinct goals: (1) to provide inspiration and focus to research on CVT for the coming years, (2) to reinforce international collaboration, and (3) to facilitate the acquisition of research funding.Summary of review:This international research agenda is the result of a research summit organized by the International Cerebral Venous Thrombosis Consortium in Amsterdam, the Netherlands, in June 2023. The summit brought together 45 participants from 15 countries including clinical researchers from various disciplines, patients who previously suffered from CVT, and delegates from industry and non-profit funding organizations. The research agenda is categorized into six pre-specified themes: (1) epidemiology and clinical features, (2) life after CVT, (3) neuroimaging and diagnosis, (4) pathophysiology, (5) medical treatment, and (6) endovascular treatment. For each theme, we present two to four research questions, followed by a brief substantiation per question. The research questions were prioritized by the participants of the summit through consensus discussion.Conclusions:This international research agenda provides an overview of the most burning research questions on CVT. Answering these questions will advance our understanding and management of CVT, which will ultimately lead to improved outcomes for CVT patients worldwide

Influenza A Viruses from Wild Birds in Guatemala Belong to the North American Lineage

The role wild bird species play in the transmission and ecology of avian influenza virus (AIV) is well established; however, there are significant gaps in our understanding of the worldwide distribution of these viruses, specifically about the prevalence and/or significance of AIV in Central and South America. As part of an assessment of the ecology of AIV in Guatemala, we conducted active surveillance in wild birds on the Pacific and Atlantic coasts. Cloacal and tracheal swab samples taken from resident and migratory wild birds were collected from February 2007 to January 2010.1913 samples were collected and virus was detected by real time RT-PCR (rRT-PCR) in 28 swab samples from ducks (Anas discors). Virus isolation was attempted for these positive samples, and 15 isolates were obtained from the migratory duck species Blue-winged teal. The subtypes identified included H7N9, H11N2, H3N8, H5N3, H8N4, and H5N4. Phylogenetic analysis of the viral sequences revealed that AIV isolates are highly similar to viruses from the North American lineage suggesting that bird migration dictates the ecology of these viruses in the Guatemalan bird population