Diagnosis and Characterization of Plant Viruses Using HTS to Support Virus Management and Tomato Breeding.

Viral diseases pose a significant threat to tomato crops (Solanum lycopersicum L.), one of the world's most economically important vegetable crops. The limited genetic diversity of cultivated tomatoes contributes to their high susceptibility to viral infections. To address this challenge, tomato breeding programs must harness the genetic resources found in native populations and wild relatives. Breeding efforts may aim to develop broad-spectrum resistance against the virome. To identify the viruses naturally infecting 19 advanced lines, derived from native tomatoes, high-throughput sequencing (HTS) of small RNAs and confirmation with PCR and RT-PCR were used. Single and mixed infections with tomato mosaic virus (ToMV), tomato golden mosaic virus (ToGMoV), and pepper huasteco yellow vein virus (PHYVV) were detected. The complete consensus genomes of three variants of Mexican ToMV isolates were reconstructed, potentially forming a new ToMV clade with a distinct 3' UTR. The absence of reported mutations associated with resistance-breaking to ToMV suggests that the Tm-1, Tm-2, and Tm-22 genes could theoretically be used to confer resistance. However, the high mutation rates and a 63 nucleotide insertion in the 3' UTR, as well as amino acid mutations in the ORFs encoding 126 KDa, 183 KDa, and MP of Mexican ToMV isolates, suggest that it is necessary to evaluate the capacity of these variants to overcome Tm-1, Tm-2, and Tm-22 resistance genes. This evaluation, along with the characterization of advanced lines using molecular markers linked to these resistant genes, will be addressed in future studies as part of the breeding strategy. This study emphasizes the importance of using HTS for accurate identification and characterization of plant viruses that naturally infect tomato germplasm based on the consensus genome sequences. This study provides crucial insights to select appropriate disease management strategies and resistance genes and guide breeding efforts toward the development of virus-resistant tomato varieties.

Polyphenolic Characterization and Anti-Inflammatory Effect of In Vitro Digested Extracts of Echinacea purpurea L. Plant Parts in an Inflammatory Model of Human Colon Cells.

Echinacea purpurea L. (EP) preparations are globally popular herbal supplements known for their medicinal benefits, including anti-inflammatory activities, partly related to their phenolic composition. However, regarding their use for the management of inflammation-related intestinal diseases, the knowledge about the fate of orally ingested constituents throughout the human gastrointestinal tract and the exposition of in vitro digested extracts in relevant inflammatory models are unknown. This study investigated for the first time the impact of in vitro gastrointestinal digestion (INFOGEST) on the phenolic composition and anti-inflammatory properties of EP extracts from flowers (EF), leaves (EL), and roots (ER) on IL-1ß-treated human colon-derived CCD-18Co cells. Among the seven hydroxycinnamic acids identified using HPLC-UV-MS/MS, chicoric and caftaric acids showed the highest concentrations in EL, followed by EF and ER, and all extracts exerted significant reductions in IL-6, IL-8, and PGE2 levels. After digestion, despite reducing the bioaccessibility of their phenolics, the anti-inflammatory effects were preserved for digested EL and, to a lesser extent, for EF, but not for digested ER. The lower phenolic content in digested EF and ER could explain these findings. Overall, this study emphasizes the potential of EP in alleviating intestinal inflammatory conditions and related disorders.

Conventional office blood pressure measurements and unattended automated office blood pressure compared with home self-measurement and 24-h ambulatory blood pressure monitoring.

OBJECTIVE: To assess whether automated office blood pressure (BP) (AOBP) measurement is a better method for measuring BP in the office than conventional techniques and an alternative to out-of-office BP measurements: home-self BP (HSBP) or ambulatory BP monitoring (ABPM). METHODS: We conducted a cross-sectional study of 74 patients and compared AOBP with the conventional technique using a mercury sphygmomanometer and with both out-to-office BP measurements: HSBP of 7 days (three measurements in the morning, afternoon, and night) and daytime ABPM. In addition, we compared BP values obtained using HSBP and ABPM to determine their level of agreement. We used ANOVA to compare means, Bland-Altman, and intraclass correlation coefficients (ICC) for concordance. RESULTS: BP values obtained by the two office methods were similar: conventional 147.2/85.0 mmHg and AOBP 146.0/85.5 mmHg ( P > 0.05) with good agreement (ICC 0.85). The mean SBP differences between AOBP and HSBP ( P < 0.001) and between AOBP and ABPM ( P < 0.001) were 8.6/13.0 mmHg with limits of agreement of -21.2 to 38.5 and -18.4 to 44.3 mmHg, respectively. The average SBP values obtained by HSBP were 4.3 mmHg higher than those obtained by ABPM ( P < 0.01). CONCLUSION: Our study showed good agreement and concordance between the two office methods as well between the two out-to-office methods, although there was a significant difference in the mean SBP between the HSBP and ABPM. Moreover, AOBP was not comparable to either HSBP or ABPM; therefore, the estimation of out-to-office BP using AOBP is not supported.

Main Determinants Affecting the Antiproliferative Activity of Stilbenes and Their Gut Microbiota Metabolites in Colon Cancer Cells: A Structure-Activity Relationship Study.

trans-Resveratrol can be catabolized by the gut microbiota to dihydroresveratrol, 3,4'-dihydroxy-trans-stilbene, lunularin, and 4-hydroxydibenzyl. These metabolites can reach relevant concentrations in the colon. However, not all individuals metabolize RSV equally, as it depends on their RSV gut microbiota metabotype (i.e., lunularin producers vs. non-producers). However, how this microbial metabolism affects the cancer chemopreventive activity of stilbenes and their microbial metabolites is poorly known. We investigated the structure-antiproliferative activity relationship of dietary stilbenes, their gut microbial metabolites, and various analogs in human cancer (Caco-2 and HT-29) and non-tumorigenic (CCD18-Co) colon cells. The antiproliferative IC50 values of pterostilbene, oxy-resveratrol, piceatannol, resveratrol, dihydroresveratrol, lunularin, 3,4'-dihydroxy-trans-stilbene, pinosylvin, dihydropinosylvin, 4-hydroxy-trans-stilbene, 4-hydroxydibenzyl, 3-hydroxydibenzyl, and 4-trans-stilbenemethanol were calculated. IC50 values were correlated with 34 molecular characteristics by bi- and multivariate analysis. Little or no activity on CCD18-Co was observed, while Caco-2 was more sensitive than HT-29, which was explained by their different capacities to metabolize the compounds. Caco-2 IC50 values ranged from 11.4 ± 10.1 µM (4-hydroxy-trans-stilbene) to 73.9 ± 13.8 µM (dihydropinosylvin). In HT-29, the values ranged from 24.4 ± 11.3 µM (4-hydroxy-trans-stilbene) to 96.7 ± 6.7 µM (4-hydroxydibenzyl). At their IC50, most compounds induced apoptosis and arrested the cell cycle at the S phase, pterostilbene at G2/M, while 4-hydroxy-trans-stilbene and 3,4'-dihydroxy-trans-stilbene arrested at both phases. Higher Connolly values (larger size) hindered the antiproliferative activity, while a lower pKa1 enhanced the activity in Caco-2, and higher LogP values (more hydrophobicity) increased the activity in HT-29. Reducing the styrene double bond in stilbenes was the most critical feature in decreasing the antiproliferative activity. These results (i) suggest that gut microbiota metabolism determines the antiproliferative effects of dietary stilbenes. Therefore, RSV consumption might exert different effects in individuals depending on their gut microbiota metabotypes associated with RSV metabolism, and (ii) could help design customized drugs with a stilbenoid and (or) dibenzyl core against colorectal cancer.

ADAM17/MMP inhibition prevents neutrophilia and lung injury in a mouse model of COVID-19.

Severe coronavirus disease 2019 (COVID-19) is characterized by lung injury, cytokine storm, and increased neutrophil-to-lymphocyte ratio (NLR). Current therapies focus on reducing viral replication and inflammatory responses, but no specific treatment exists to prevent the development of severe COVID-19 in infected individuals. Angiotensin-converting enzyme-2 (ACE2) is the receptor for SARS-CoV-2, the virus causing COVID-19, but it is also critical for maintaining the correct functionality of lung epithelium and endothelium. Coronaviruses induce activation of a disintegrin and metalloprotease 17 (ADAM17) and shedding of ACE2 from the cell surface resulting in exacerbated inflammatory responses. Thus, we hypothesized that ADAM17 inhibition ameliorates COVID-19-related lung inflammation. We employed a preclinical mouse model using intratracheal instillation of a combination of polyinosinic:polycytidylic acid (poly(I:C)) and the receptor-binding domain of the SARS-CoV-2 spike protein (RBD-S) to mimic lung damage associated with COVID-19. Histologic analysis of inflamed mice confirmed the expected signs of lung injury including edema, fibrosis, vascular congestion, and leukocyte infiltration. Moreover, inflamed mice also showed an increased NLR as observed in critically ill COVID-19 patients. Administration of the ADAM17/MMP inhibitors apratastat and TMI-1 significantly improved lung histology and prevented leukocyte infiltration. Reduced leukocyte recruitment could be explained by reduced production of proinflammatory cytokines and lower levels of the endothelial adhesion molecules ICAM-1 and VCAM-1. Additionally, the NLR was significantly reduced by ADAM17/MMP inhibition. Thus, we propose inhibition of ADAM17/MMP as a novel promising treatment strategy in SARS-CoV-2-infected individuals to prevent the progression toward severe COVID-19.

Deep-learning based detection of COVID-19 using lung ultrasound imagery.

BACKGROUND: The COVID-19 pandemic has exposed the vulnerability of healthcare services worldwide, especially in underdeveloped countries. There is a clear need to develop novel computer-assisted diagnosis tools to provide rapid and cost-effective screening in places where massive traditional testing is not feasible. Lung ultrasound is a portable, easy to disinfect, low cost and non-invasive tool that can be used to identify lung diseases. Computer-assisted analysis of lung ultrasound imagery is a relatively recent approach that has shown great potential for diagnosing pulmonary conditions, being a viable alternative for screening and diagnosing COVID-19. OBJECTIVE: To evaluate and compare the performance of deep-learning techniques for detecting COVID-19 infections from lung ultrasound imagery. METHODS: We adapted different pre-trained deep learning architectures, including VGG19, InceptionV3, Xception, and ResNet50. We used the publicly available POCUS dataset comprising 3326 lung ultrasound frames of healthy, COVID-19, and pneumonia patients for training and fine-tuning. We conducted two experiments considering three classes (COVID-19, pneumonia, and healthy) and two classes (COVID-19 versus pneumonia and COVID-19 versus non-COVID-19) of predictive models. The obtained results were also compared with the POCOVID-net model. For performance evaluation, we calculated per-class classification metrics (Precision, Recall, and F1-score) and overall metrics (Accuracy, Balanced Accuracy, and Area Under the Receiver Operating Characteristic Curve). Lastly, we performed a statistical analysis of performance results using ANOVA and Friedman tests followed by post-hoc analysis using the Wilcoxon signed-rank test with the Holm's step-down correction. RESULTS: InceptionV3 network achieved the best average accuracy (89.1%), balanced accuracy (89.3%), and area under the receiver operating curve (97.1%) for COVID-19 detection from bacterial pneumonia and healthy lung ultrasound data. The ANOVA and Friedman tests found statistically significant performance differences between models for accuracy, balanced accuracy and area under the receiver operating curve. Post-hoc analysis showed statistically significant differences between the performance obtained with the InceptionV3-based model and POCOVID-net, VGG19-, and ResNet50-based models. No statistically significant differences were found in the performance obtained with InceptionV3- and Xception-based models. CONCLUSIONS: Deep learning techniques for computer-assisted analysis of lung ultrasound imagery provide a promising avenue for COVID-19 screening and diagnosis. Particularly, we found that the InceptionV3 network provides the most promising predictive results from all AI-based techniques evaluated in this work. InceptionV3- and Xception-based models can be used to further develop a viable computer-assisted screening tool for COVID-19 based on ultrasound imagery.

A Fluorescence Dequenching-based Liposome Leakage Assay to Measure Membrane Permeabilization by Pore-forming Proteins.

Pore-forming toxins (PFTs) have been discovered in a wide range of organisms. Their functions are essential to the survival or virulence of many species. PFTs often interact with lipid membranes. Large unilamellar vesicles (LUV), also known as liposomes, have been commonly used as reliable membrane models for testing PFTs activity. Liposomes have great adaptability in size, lipid composition, and loading cargo. Incorporating the fluorescent dye/quencher pair, 8-Aminonaphthalene-1,3,6-Trisulfonic Acid (ANTS) and p-Xylene-Bis-Pyridinium Bromide (DPX), in liposomes is an effective approach for measuring membrane leakage. When ANTS and DPX are encapsulated in a liposome, the fluorescence of ANTS is quenched by DPX. However, disruption of liposome integrity and subsequent leakage result in measurable fluorescence emitted by ANTS. Here, we report our protocol for optimal liposome preparation for measuring liposome leakage by fluorescence dequenching.

Acute Lymphoblastic Leukemia Cell Lines in Immunology Research.

A considerable portion of our knowledge on T and B cell biology is acquired from research using acute lymphoblastic leukemia (ALL) cell lines, which are invaluable tools used in many immunology and leukemia studies. Here, we discuss the advantages and limitations of ALL cell lines and provide guidelines on their proper usage.

Development of an In Vitro Membrane Model to Study the Function of EsxAB Heterodimer and Establish the Role of EsxB in Membrane Permeabilizing Activity of Mycobacterium tuberculosis.

EsxA and EsxB are secreted as a heterodimer and have been shown to play critical roles in phagosome rupture and translocation of Mycobacterium tuberculosis into the cytosol. Recent in vitro studies have suggested that the EsxAB heterodimer is dissociated upon acidification, which might allow EsxA insertion into lipid membranes. While the membrane permeabilizing activity (MPA) of EsxA has been well characterized in liposomes composed of di-oleoyl-phosphatidylcholine (DOPC), the MPA of EsxAB heterodimer has not been detected through in vitro assays due to its negligible activity with DOPC liposomes. In this study, we established a new in vitro membrane assay to test the MPA activity of N-terminal acetylated EsxA (N-EsxA). We established that a dose-dependent increase in anionic charged lipids enhances the MPA of N-EsxA. The MPA of both N-EsxA and EsxAB were significantly increased with this new liposome system and made it possible to characterize the MPA of EsxAB in more physiologically-relevant conditions. We tested, for the first time, the effect of temperature on the MPA of N-EsxA and EsxAB in this new system. Interestingly, the MPA of N-EsxA was lower at 37 °C than at RT, and on the contrary, the MPA of EsxAB was higher at 37 °C than at RT. Surprisingly, after incubation at 37 °C, the MPA of N-EsxA continuously decreased over time, while MPA of EsxAB remained stable, suggesting EsxB plays a key role in stabilizing N-EsxA to preserve its MPA at 37 °C. In summary, this study established a new in vitro model system that characterizes the MPA of EsxAB and the role of EsxB at physiological-relevant conditions.

Kv1.3 Current Voltage Dependence in Lymphocytes is Modulated by Co-Culture with Bone Marrow-Derived Stromal Cells: B and T Cells Respond Differentially.

BACKGROUND/AIMS: Kv1.3 channel is the only voltage-dependent potassium channel in plasma membrane of human lymphocytes. Bearing in mind a rather steep voltage-dependence of Kv1.3 activation and inactivation, its modulation by B and T cells activation and by co-culture with stromal bone-marrow cells was addressed. METHODS: Patch-clamp technique in the whole cell mode was applied to human resting and activated human B and T cells, in monoculture and co-culture with stromal OP9 cells. RESULTS: Polyclonal activation of B and T cells in monoculture caused Kv1.3 current in B cells to activate at more negative and in T cells at more positive potentials, whereas the inactivation of Kv1.3 current in resting T cells occurred at more negative voltages. Co-culture with OP9 cells abolished the shift of voltage dependence upon the polyclonal activation but fixed the substantial difference between B and T cells, resting or activated, with both activation and inactivation negatively shifted by 15 mV for T lymphocytes. However, activated B cells displayed an incomplete inactivation, which was augmented by the co-culture. Neither activation nor co-culture caused substantial changes in the Kv1.3 current density. CONCLUSION: The combination of activation and inactivation processes yields the fraction of steady-state Kv1.3 current (window current), which was higher in activated B cells, partly due to an incomplete inactivation. A relatively smaller window current in resting B cells and resting T cells in co-culture correlated with a more depolarized resting membrane potential. Rather than insignificant changes in the Kv1.3 channels functional expression, the modulation of their voltage dependence by activation and co-culture with bone-marrow stromal cells was essential for the control of membrane potential.

Nα-Acetylation of the virulence factor EsxA is required for mycobacterial cytosolic translocation and virulence.

The Mycobacterium tuberculosis virulence factor EsxA and its chaperone EsxB are secreted as a heterodimer (EsxA:B) and are crucial for mycobacterial escape from phagosomes and cytosolic translocation. Current findings support the idea that for EsxA to interact with host membranes, EsxA must dissociate from EsxB at low pH. However, the molecular mechanism by which the EsxA:B heterodimer separates is not clear. In the present study, using liposome-leakage and cytotoxicity assays, LC-MS/MS-based proteomics, and CCF-4 FRET analysis, we obtained evidence that the Nα-acetylation of the Thr-2 residue on EsxA, a post-translational modification that is present in mycobacteria but absent in Escherichia coli, is required for the EsxA:B separation. Substitutions at Thr-2 that precluded Nα-acetylation inhibited the heterodimer separation and hence prevented EsxA from interacting with the host membrane, resulting in attenuated mycobacterial cytosolic translocation and virulence. Molecular dynamics simulations revealed that at low pH, the Nα-acetylated Thr-2 makes direct and frequent "bind-and-release" contacts with EsxB, which generates a force that pulls EsxB away from EsxA. In summary, our findings provide evidence that the Nα-acetylation at Thr-2 of EsxA facilitates dissociation of the EsxA:B heterodimer required for EsxA membrane permeabilization and mycobacterial cytosolic translocation and virulence.

Phase III Trial of Adjuvant Capecitabine After Standard Neo-/Adjuvant Chemotherapy in Patients With Early Triple-Negative Breast Cancer (GEICAM/2003-11_CIBOMA/2004-01).

PURPOSE: Operable triple-negative breast cancers (TNBCs) have a higher risk of relapse than non-TNBCs with standard therapy. The GEICAM/2003-11_CIBOMA/2004-01 trial explored extended adjuvant capecitabine after completion of standard chemotherapy in patients with early TNBC. PATIENTS AND METHODS: Eligible patients were those with operable, node-positive-or node negative with tumor 1 cm or greater-TNBC, with prior anthracycline- and/or taxane-containing chemotherapy. After central confirmation of TNBC status by immunohistochemistry, patients were randomly assigned to either capecitabine or observation. Stratification factors included institution, prior taxane-based therapy, involved axillary lymph nodes, and centrally determined phenotype (basal v nonbasal, according to cytokeratins 5/6 and/or epidermal growth factor receptor positivity by immunohistochemistry). The primary objective was to compare disease-free survival (DFS) between both arms. RESULTS: Eight hundred seventy-six patients were randomly assigned to capecitabine (n = 448) or observation (n = 428). Median age was 49 years, 55.9% were lymph node negative, 73.9% had a basal phenotype, and 67.5% received previous anthracyclines plus taxanes. Median length of follow-up was 7.3 years. DFS was not significantly prolonged with capecitabine versus observation [hazard ratio (HR), 0.82; 95% CI, 0.63 to 1.06; P = .136]. In a preplanned subgroup analysis, nonbasal patients seemed to derive benefit from the addition of capecitabine with a DFS HR of 0.53 versus 0.94 in those with basal phenotype (interaction test P = .0694) and an HR for overall survival of 0.42 versus 1.23 in basal phenotype (interaction test P = .0052). Tolerance of capecitabine was as expected, with 75.2% of patients completing the planned 8 cycles. CONCLUSION: This study failed to show a statistically significant increase in DFS by adding extended capecitabine to standard chemotherapy in patients with early TNBC. In a preplanned subset analysis, patients with nonbasal phenotype seemed to obtain benefit with capecitabine, although this will require additional validation.

Effects of membrane lipid composition on Mycobacterium tuberculosis EsxA membrane insertion: A dual play of fluidity and charge.

As a key virulence factor of Mycobacterium tuberculosis, EsxA or 6-kDa early secreted antigenic target (ESAT-6) has been implicated in phagosome rupture and mycobacterial translocation from the phagosome to the cytosol within macrophages. Our previous studies have shown that EsxA permeabilizes liposomal membrane at acidic pH and a membrane-permeabilization defective mutant Q5K attenuates mycobacterial cytosolic translocation and virulence in macrophages. To further probe the mechanism of EsxA membrane permeabilization, here we characterized the effects of various lipid compositions, including biologically relevant phagosome-mimicking lipids and lipid rafts, on the structural stability and membrane insertion of EsxA WT and Q5K. We have found a complex dual play of membrane fluidity and charge in regulating EsxA membrane insertion. Moreover, Q5K affects the membrane insertion through a structure- and lipid composition-independent mechanism. The results of this study provide a novel insights into the mechanism of EsxA membrane interaction.

Impacto del índice de masa corporal en la presión arterial medida con esfigmomanómetro de mercurio en niños y adolescentes con diabetes mellitus tipo 1 / The impact of body mass index on blood pressure measured with mercury sphygmomanometer in children and adolescents with type 1 diabetes mellitus

Resumen Introducción Los pacientes con diabetes mellitus tipo 1 (DM1) y sobrepeso tienen más riesgo de desarrollar cambios en la presión arterial (PA), y esto incrementa su morbilidad y mortalidad cardiovascular. En este estudio se determinó la relación entre la PA y el índice de masa corporal (IMC) y el promedio de las tres últimas mediciones de hemoglobina glucosilada (HbA1c) de pacientes con DM1. Métodos Estudio transversal analítico en niños y adolescentes con DM1 con más de un año de evolución. Las variables dependientes fueron la PA sistólica y diastólica medidas con esfigmomanómetro y las variables independientes, IMC y promedio de las últimas tres mediciones de la HbA1c. Se utilizó regresión lineal múltiple con intervalo de confianza del 95%. Resultados Se estudiaron 75 pacientes con DM1. La mediana del tiempo de evolución de la DM1 fue de 3.5 años (mínimo 1 año-máximo 14.8 años), el IMC 19.5 ± 3.1 kg/cm2 y la HbA1c 8.3 ± 2.4%. De los 75 pacientes, 66 presentaron PA < percentil 90 y 9 PA ≥ percentil 90 (12%). Se construyeron dos modelos de regresión lineal múltiple, con PA sistólica y diastólica como variables dependientes. Las posibles variables predictoras fueron sugeridas por el contexto teórico y el análisis estadístico. El IMC expresado en puntuación zeta (zIMC) fue predictor para PA sistólica/diastólica. Los modelos sugirieron que a cada incremento de unidad del zIMC corresponde un aumento de 5.1 y 3.6 mmHg de PA sistólica y diastólica, respectivamente. Conclusiones Se observó una correlación positiva de la PA sistólica y la diastólica con el zIMC.

Abstract Background Patients with type 1 diabetes mellitus (T1DM) and overweight have more risk to develop changes in blood pressure that increase cardiovascular morbidity and mortality. In this study, the relationship between blood pressure (BP) with the body mass index (BMI) and the average of the last three measurements of glycated hemoglobin (HbA1c) in patients with T1DM was determined. Methods A cross-sectional analytical study was conducted in children and adolescents with T1DM with over a year since diagnosis. The dependent variables were systolic and diastolic BP, measured with a mercury sphygmomanometer. The independent variables were BMI and average of the last three measurements of HbA1. A linear regression with a 95% confidence interval was used. Results Seventy-five patients with T1DM were studied. The median of disease duration was 3.5 years (min 1-max 14.8 years), BMI 19.5 ± 3.1 kg/cm2 and HbA1c 8.3 ± 2.4%. Sixty-six patients showed BP < percentile 90 and 9 BP ≥ percentile 90 (12%). Two models of linear regression were constructed, with systolic and diastolic BP as dependent variables. The possible predictor variables were suggested by theoretical context and statistical analysis. The predictive variable of high BP was zBMI (body mass index expressed in z-score) for systolic and diastolic BP. Also, the models suggested that for an increase of one unit of zBMI, corresponded a rise of 5.1 and 3.6 mmHg in systolic and diastolic BP, respectively. Conclusions A positive correlation between systolic and diastolic BP with zBMI was observed.

The impact of body mass index on blood pressure measured with mercury sphygmomanometer in children and adolescents with type 1 diabetes mellitus.

Background: Patients with type 1 diabetes mellitus (T1DM) and overweight have more risk to develop changes in blood pressure that increase cardiovascular morbidity and mortality. In this study, the relationship between blood pressure (BP) with the body mass index (BMI) and the average of the last three measurements of glycated hemoglobin (HbA1c) in patients with T1DM was determined. Methods: A cross-sectional analytical study was conducted in children and adolescents with T1DM with over a year since diagnosis. The dependent variables were systolic and diastolic BP, measured with a mercury sphygmomanometer. The independent variables were BMI and average of the last three measurements of HbA1. A linear regression with a 95% confidence interval was used. Results: Seventy-five patients with T1DM were studied. The median of disease duration was 3.5 years (min 1-max 14.8 years), BMI 19.5 ± 3.1 kg/cm2 and HbA1c 8.3 ± 2.4%. Sixty-six patients showed BP < percentile 90 and 9 BP ≥ percentile 90 (12%). Two models of linear regression were constructed, with systolic and diastolic BP as dependent variables. The possible predictor variables were suggested by theoretical context and statistical analysis. The predictive variable of high BP was zBMI (body mass index expressed in z-score) for systolic and diastolic BP. Also, the models suggested that for an increase of one unit of zBMI, corresponded a rise of 5.1 and 3.6 mmHg in systolic and diastolic BP, respectively. Conclusions: A positive correlation between systolic and diastolic BP with zBMI was observed.

Introducción: Los pacientes con diabetes mellitus tipo 1 (DM1) y sobrepeso tienen más riesgo de desarrollar cambios en la presión arterial (PA), y esto incrementa su morbilidad y mortalidad cardiovascular. En este estudio se determinó la relación entre la PA y el índice de masa corporal (IMC) y el promedio de las tres últimas mediciones de hemoglobina glucosilada (HbA1c) de pacientes con DM1. Métodos: Estudio transversal analítico en niños y adolescentes con DM1 con más de un año de evolución. Las variables dependientes fueron la PA sistólica y diastólica medidas con esfigmomanómetro y las variables independientes, IMC y promedio de las últimas tres mediciones de la HbA1c. Se utilizó regresión lineal múltiple con intervalo de confianza del 95%. Resultados: Se estudiaron 75 pacientes con DM1. La mediana del tiempo de evolución de la DM1 fue de 3.5 años (mínimo 1 año-máximo 14.8 años), el IMC 19.5 ± 3.1 kg/cm2 y la HbA1c 8.3 ± 2.4%. De los 75 pacientes, 66 presentaron PA < percentil 90 y 9 PA ≥ percentil 90 (12%). Se construyeron dos modelos de regresión lineal múltiple, con PA sistólica y diastólica como variables dependientes. Las posibles variables predictoras fueron sugeridas por el contexto teórico y el análisis estadístico. El IMC expresado en puntuación zeta (zIMC) fue predictor para PA sistólica/diastólica. Los modelos sugirieron que a cada incremento de unidad del zIMC corresponde un aumento de 5.1 y 3.6 mmHg de PA sistólica y diastólica, respectivamente. Conclusiones: Se observó una correlación positiva de la PA sistólica y la diastólica con el zIMC.

Differential Activity of Voltage- and Ca2+-Dependent Potassium Channels in Leukemic T Cell Lines: Jurkat Cells Represent an Exceptional Case.

Activation of resting T cells relies on sustained Ca2+ influx across the plasma membrane, which in turn depends on the functional expression of potassium channels, whose activity repolarizes the membrane potential. Depending on the T-cells subset, upon activation the expression of Ca2+- or voltage-activated K+ channels, KCa or Kv, is up-regulated. In this study, by means of patch-clamp technique in the whole cell mode, we have studied in detail the characteristics of Kv and KCa currents in resting and activated human T cells, the only well explored human T-leukemic cell line Jurkat, and two additional human leukemic T cell lines, CEM and MOLT-3. Voltage dependence of activation and inactivation of Kv1.3 current were shifted up to by 15 mV to more negative potentials upon a prolonged incubation in the whole cell mode and displayed little difference at a stable state in all cell lines but CEM, where the activation curve was biphasic, with a high and low potential components. In Jurkat, KCa currents were dominated by apamine-sensitive KCa2.2 channels, whereas only KCa3.1 current was detected in healthy T and leukemic CEM and MOLT-3 cells. Despite a high proliferation potential of Jurkat cells, Kv and KCa currents were unexpectedly small, more than 10-fold lesser as compared to activated healthy human T cells, CEM and MOLT-3, which displayed characteristic Kv1.3high:KCa3.1high phenotype. Our results suggest that Jurkat cells represent perhaps a singular case and call for more extensive studies on primary leukemic T cell lines as well as a verification of the therapeutic potential of specific KCa3.1 blockers to combat acute lymphoblastic T leukemias.

Comparison of auscultatory and oscillometric BP measurements in children with obesity and their effect on the diagnosis of arterial hypertension / Comparación de la medición de PA auscultatoria y oscilométrica en niños con obesidad y su efecto en el diagnóstico de hipertensión arterial

Abstract: Objective: The level of agreement between two blood pressure (BP) reading methods, auscultatory vs oscillometric, was examined using a mercury sphygmomanometer and an electronic device in children and adolescents with different levels of obesity. The readings were compared to determine their impact on the diagnosis of pre-hypertension/hypertension. Methods: Blood pressure readings were taken in children with obesity (body mass index ≥ 95th percentile) and severe obesity (≥120% 95th percentile). Bland-Altman analysis and Intraclass Correlation Coefficient were used to determine the agreement between measurements. Results: The mercury sphygmomanometer readings were lower than those obtained with the electronic device for both systolic and diastolic BP (P = .01 and P = .001, respectively). The mean systolic and diastolic BP differences between the oscillometric vs first mercury reading were 4.2/10.2 mm Hg, respectively. A large difference was observed between the BP measurement methods. The ICC showed regular to moderate reliability for the systolic BP (.595), but poor for the diastolic BP (.330). Screening using the first of three mercury measurements showed that 10.4% of the children and adolescents had BPs within the pre-hypertension/hypertension range. This was reduced to 5.2% when the mean of three mercury readings was used. Conclusions: Large discrepancies were observed in both the systolic and diastolic BP. These differences are not clinically acceptable as to consider the two instruments interchangeable. The electronic device readings were higher, and they overestimated the diagnosis of hypertension. © 2017 Instituto Nacional de Cardiología Ignacio Chávez. Published by Masson Doyma Mèxico S.A. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Resumen: Objetivo: Para conocer el grado de concordancia entre 2 métodos de medición de presión arterial (PA), auscultatorio vs oscilométrico se utilizó un esfigmomanómetro de mercurio y un dispositivo electrónico en niños y adolescentes con diferentes grados de obesidad. Las lecturas fueron comparadas para conocer su impacto en el diagnóstico de prehipertensión/hipertensión. Método: Se midió la PA a niños con obesidad (percentil 95 del índice masa corporal) y obesidad severa (120% del percentil 95). Utilizamos análisis de Bland-Altman y Coeficiente de Correlación Intraclase (CCI) para conocer acuerdo entre mediciones. Resultados: Las lecturas con esfigmomanómetro de mercurio fueron más bajas que con el electrónico para la PA sistólica y diastólica (p = 0.01 y 0.001, respectivamente). El promedio de las diferencias en sistólica y diastólica entre oscilométrico vs. primera medición con mercurio fue de 4.2/10.2 mm Hg respectivamente. Se observó una gran diferencia de las mediciones entre los métodos de medición de PA. El CCI mostró una confiabilidad regular a moderada para la sistólica (0.595) pero pobre para la diastólica (0.330). El tamizaje con una medición mediante mercurio mostró que el 10.4% de los niños y adolescentes tenían PA en el rango de prehipertensión/hipertensión, pero se redujo a un 5.2% con el promedio de 3 mediciones. Conclusiones: Se observaron grandes discrepancias en la PA sistólica y diastólica. Tales diferencias no son clínicamente aceptables como para considerar equivalentes los 2 instrumentos. Las mediciones realizadas en este estudio con dispositivo electrónico fueron más altas y sobre estimaron el diagnóstico de hipertensión. © 2017 Instituto Nacional de Cardiología Ignacio Chávez. Publicado por Masson Doyma México S.A. Este es un artículo Open Access bajo la licencia CC BY-NC-ND (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Wavefronts and caustics associated with Mathieu beams.

In this work we compute the wavefronts and the caustics associated with the solutions to the scalar wave equation introduced by Durnin in elliptical cylindrical coordinates generated by the function A(ϕ)=ceν(ϕ,q)+iseν(ϕ,q), with ν being an integral or nonintegral number. We show that the wavefronts and the caustic are invariant under translations along the direction of evolution of the beam. We remark that the wavefronts of the separable Mathieu beams generated by A(ϕ)=ceν(ϕ,q) and A(ϕ)=seν(ϕ,q) are cones and their caustic is the z axis; thus, they are not structurally stable. However, in general, the Mathieu beam generated by A(ϕ)=ceν(ϕ,q)+iseν(ϕ,q) is stable because locally its caustic has singularities of the fold and cusp types. To show this property, we present the wavefronts and the caustics for the Mathieu beams with characteristic value aν=0 and q=0,0.2,0.3,0.5. For q=0, we obtain the Bessel beam of order zero; in this case, the wavefronts are cones and the caustic coincides with the z axis. For q≠0, the wavefronts are deformations of conical ones, and the caustic surface, for some values of q, has singularities of the cusp ridge type. Furthermore, we remark that the set of Mathieu beams with characteristic value aν=0 and 0≤q<1 has associated a caustic with singularities of the swallowtail type, which is structurally stable. Therefore, we conclude that this type of Mathieu beam is more stable than plane waves, Bessel beams, parabolic beams, and those generated by A(ϕ)=ceν(ϕ,q) and A(ϕ)=seν(ϕ,q). To support this conclusion, we present experimental results showing the pattern obtained after obstructing a plane wave, the Bessel beam of order m=5, and the Mathieu beam of order m=5 and q=50 with complex transversal amplitude given by Ce5(ξ,50)ce5(η,50)+iSe5(ξ,50)se5(η,50), where (ξ, η) are the elliptical coordinates on the plane.