Article biodiversity inside bottles: animals, fungi, and plants in traditional alcoholic drinks.

The use of animals, fungi, and plants as a source of bioactive compounds has been widely practiced in diverse cultures throughout the world, particularly in alcoholic drinks. The nature of the biological material, method of preparation and alcohol concentration play a predominant role in the extraction of bioactive compounds and the achievement of desired results. However, certain aspects must be considered to guarantee the innocuity of these drinks and reduce the risk of intoxication, infections and allergic reactions, aspects which are sometimes overlooked. In addition, the implications of using threatened or protected species must be considered to reduce the negative impact on their populations. The authors recommend the establishment of production systems which guarantee products with adequate quality controls and ensure the benefits to the consumer.

Phenotypic Expression and Outcomes in Patients with the p.Arg301Gln GLA Variant in Anderson-Fabry Disease.

The p.Arg301Gln variant in the α -galactosidase A gene (GLA) has been poorly described in the literature. The few reports show controversial information, with both classical and nonclassical Anderson-Fabry Disease (AFD) presentation patterns. The aim of this study was to analyze the penetrance, clinical phenotype, and biochemical profile of an international cohort of patients carrying the p.Arg301Gln genetic variant in the GLA gene. This was an observational, international, and retrospective cohort case series study of patients carrying the p.Arg301Gln variant in the GLA gene associated with AFD disease. Forty-nine p.Arg301Gln GLA carriers, 41% male, were analyzed. The penetrance was 63% in the entire cohort and 1.5 times higher in men. The mean age of symptoms onset was 41 years; compared to women, men presented symptoms earlier and with a shorter delay to diagnosis. The typical clinical triad-cornea verticillate, neuropathic pain, and angiokeratomas-affected only 20% of the cohort, with no differences between genders. During follow-up, almost 20% of the patients presented some type of nonfatal cardiovascular and renal event (stroke, need for dialysis, heart failure, and arrhythmias requiring intracardiac devices), predominantly affecting men. Residual levels were the most common finding of α-GAL A enzyme activity, only a few women had a normal level; a small proportion of men had undetectable levels. The incidence of combined outcomes including all causes of death was 33%, and the cumulative incidence of all-cause mortality was 9% at the follow-up. Patients carrying the p.Arg301Gln GLA variant have a high penetrance, with predominantly cardiorenal involvement and clinical onset of the disease in middle age. Only a small proportion showed the classic clinical presentation of AFD. As in other X-linked diseases, males were more affected by severe cardiovascular and renal events. This genotype-phenotype correlation could be useful from a practical clinical point of view and for future decision making.

Piquin chili, a wild spice: natural variation in nutraceutical contents.

The piquin chili is a wild spice widely consumed from the South United States to Central America and stands out as a source of flavonoids, essential metabolites with antioxidant properties. The concentrations of flavonoids, carotenoids, and capsaicinoids vary according to regions, maturity stages, and ripening processes. These compounds, which are known for their health benefits and industrial applications, highlight the importance of identifying ideal environmental conditions for collecting fruits with the highest contents. Comprehensive studies of the piquin chili are essential for understanding its properties for the benefit of consumers. This approach fortifies trade, contributes to resource conservation, and advances cultivated chili production.

Largest comparison between onset and relapses of acquired thrombotic thrombocytopenic purpura reveals severe neurological involvement and worse analytic parameters at debut.

It has been proposed that the onset of Acquired Thrombotic Thrombocytopenic Purpura (iTTP) is more severe than subsequent relapses; however, existing studies have limitations. We conducted a retrospective observational study to compare analytical and clinical severity of onset and relapse aTTP cases between 2012 and 2023. A total of 370 episodes of aTTP were analyzed, comprising 272 at initial diagnosis and 98 relapses. At onset, analytical parameters indicative of severity (low hemoglobin, low platelet count, and increased LDH) were significantly worse; patients had severe neurological symptoms (p<0.001) and ≥ 3 points in the TMA mortality score (p<0.001). In conclusion, the onset of aTTP is associated with worse analytical parameters and severe neurological involvement.

A New Optical Interferometric Biosensing System Enhanced with Nanoparticles for Alzheimer's Disease in Serum.

In this scientific work, we demonstrate, for the first time, a new biosensing system and procedure to measure specifically the total Tau (T-Tau) protein in serum, one of the most relevant biomarkers of Alzheimer's disease (AD). AD is a progressive brain disorder that produces neuronal and cognitive dysfunction and affects a high percentage of people worldwide. For this reason, diagnosing AD at the earliest possible stage involves improving diagnostic systems. We report on the use of interferometric bio-transducers integrated with 65 microwells forming diagnostic KITs read-out by using the Interferometric Optical Detection Method (IODM). Moreover, biofunctionalized silicon dioxide (SiO2) nanoparticles (NPs) acting as interferometric enhancers of the bio-transducers signal allow for the improvement of both the optical read-out signal and its ability to work with less-invasive biological samples such as serum instead of cerebrospinal fluid (CSF). As a result, in this paper, we describe for the first time a relevant diagnostic alternative to detect Tau protein at demanding concentrations of 10 pg/mL or even better, opening the opportunity to be used for detecting other relevant AD-related biomarkers in serum, such as ß-amyloid and phosphorylated Tau (P-Tau), neurofilaments, among others that can be considered relevant for AD.

Trade-off between thermal preference and sperm maturation in a montane lizard.

Temperature is a key abiotic factor that influences performance of several physiological traits in ectotherms. Organisms regulate their body temperature within a range of temperatures to enhance physiological function. The capacity of ectotherms, such as lizards, to maintain their body temperature within their preferred range influences physiological traits such as speed, various reproductive patterns, and critical fitness components, such as growth rates or survival. Here, we evaluate the influence of temperature on locomotor performance, sperm morphology and viability in a high elevation lizard species (Sceloporus aeneus). Whereas maximal values for sprint speed coincides with field active and preferred body temperature, short-term exposure at the same range of temperatures produces abnormalities in sperm morphology, lower sperm concentration and diminishes sperm motility and viability. In conclusion, we confirmed that although locomotor performance is maximized at preferred temperatures, there is a trade-off with male reproductive attributes, which may cause infertility. As a consequence, prolonged exposure to preferred temperatures could threaten the persistence of the species through reduced fertility. Persistence of the species is favored in environments with access to cooler, thermal microhabitats that enhance reproductive parameters.

Integration of Multiple Interferometers in Highly Multiplexed Diagnostic KITs to Evaluate Several Biomarkers of COVID-19 in Serum.

In the present work, highly multiplexed diagnostic KITs based on an Interferometric Optical Detection Method (IODM) were developed to evaluate six Coronavirus Disease 2019 (COVID-19)-related biomarkers. These biomarkers of COVID-19 were evaluated in 74 serum samples from severe, moderate, and mild patients with positive polymerase chain reaction (PCR), collected at the end of March 2020 in the Hospital Clínico San Carlos, in Madrid (Spain). The developed multiplexed diagnostic KITs were biofunctionalized to simultaneously measure different types of specific biomarkers involved in COVID-19. Thus, the serum samples were investigated by measuring the total specific Immunoglobulins (sIgT), specific Immunoglobulins G (sIgG), specific Immunoglobulins M (sIgM), specific Immunoglobulins A (sIgA), all of them against SARS-CoV-2, together with two biomarkers involved in inflammatory disorders, Ferritin (FER) and C Reactive Protein (CRP). To assess the results, a Multiple Linear Regression Model (MLRM) was carried out to study the influence of IgGs, IgMs, IgAs, FER, and CRP against the total sIgTs in these serum samples with a goodness of fit of 73.01% (Adjusted R-Squared).

Evaluation of Resistance of Eleven Maize Races (Zea mays L.) to the Red Spider Mite (Tetranychus merganser, Boudreaux).

At least 59 maize races (Zea mays L.) have been registered in Mexico. The feeding damage caused by insects and mites to maize crops generates up to ~30% of maize yield losses. Spider-mite-resistant plants are needed. The red spider mite, Tetranychus merganser Boudreaux (Acari: Tetranychidae), is distributed in the United States, China, Mexico, and Thailand. It is considered a potential pest in Mexican agriculture. The aim of this study was to evaluate the resistance mechanisms (antixenosis and antibiosis) of 11 native maize populations, representative of each race of maize grown in Tamaulipas, Mexico, to T. merganser under laboratory conditions. The aim was also to obtain information on the chemical composition and some morphological characteristics of these maize races and to identify resistant maize races for incorporation into a breeding program. Antixenosis was assessed by non-preference for oviposition and feeding. Antibiosis was measured by growth rate (ri). The presence of secondary metabolites in the 11 maize races were different. In the 11 maize races, quantitative analysis of total phenol concentration, total flavonoid concentration, and antioxidant capacity were significantly different. The multivariate analysis of variance showed that there is evidence of antixenosis noted by maize race differences in egg laying and percentage feeding damage but not of antibiosis noted by growth rate. Red spider mites laid significantly more eggs on the Celaya (24 h: 25.67 ± 17.04, 48 h: 42.67 ± 26.86, 72 h: 49.33 ± 28.54) race than on Raton (24 h: 7.00 ± 5.00, 48 h: 12.67 ± 8.02, 72 h: 14.67 ± 9.29) and Elotes Occidentales × Tuxpeño (24 h: 9.67 ± 5.85, 48 h: 15.33 ± 10.69, 72 h: 17.67 ± 10.97) races. However, the growth rate and mortality of T. merganser in the 11 corn races were similar. The Vandeño (24 h: 11.67 ± 2.89, 48 h: 27.67 ± 7.64, 72 h: 30.00 ± 18.03) and Tabloncillo × Tuxpeño (24 h: 18.33 ± 7.64, 48 h: 25.00 ± 8.66, 72 h: 25.00 ± 8.66) races were the most resistant to red spider mite damage, whereas the most susceptible race was Celaya (24 h: 26.67 ± 15.28, 48 h: 48.33 ± 29.30, 72 h: 65.00 ± 30.00). Further analysis by PCA at 24, 48, and 72 h found the Celaya race positively correlated to growth rate and oviposition of T. merganser and to a lesser extent with the percentage of feeding damage, suggesting that the Celaya race was most susceptible to T. merganser. At 24 h, the Vandeño race was most resistant, given a negative correlation to growth rate and oviposition by T. merganser. The PCA at 48 and 72 h noted the Elotes Occidentales × Tuxpeño race was most resistant to red spider mite, with negative relationships to growth rate and oviposition and, to a lesser extent, to feeding damage. This resistance is due to the differences in both its morphological characteristics and the secondary metabolites present in their leaves.

New Label-Free Biosensing for the Evaluation of the AX-024 Inhibitor: Case Study for the Development of New Drugs in Autoimmune Diseases.

We developed a new label-free assay to evaluate the inhibition capacity of AX-024 by means of a new Point-of-Care (PoC) device for application in the development of new drugs in autoimmune diseases. The technology of PoC is based on interferometric optical detection method (IODM). For this purpose, we have optimized and developed an assay protocol whereby a Glutathione S-Transferase modified protein (GST-SH3.1), which contains a functional domain of a protein involved in T-cell activation, together with the AX-024 inhibitor has been studied. The chips used are a sensing surface based on nitrocellulose. We used streptavidin and a biotinylated peptide as links for the immobilization process on the sensing surface. The biotinylated peptide and AX-024 inhibitor compete for the same functional group of the GST-SH3.1 modified protein. When the inhibitor binds its binding site on GST-SH3.1, the biotinylated peptide cannot bind to its pocket on the protein. This competition reduces the total molecular mass of protein fixed onto the biosensor. In order to quantify the inhibition capacity of AX-024, several Ax-024:GST-SH3.1 ratios have been studied. We have compared the read-out signal for GST-SH3.1 protein not interfered by the drug, which served as a positive blank, and the response of the GST-SH3.1 modified protein blocked by the inhibitor. The technology has been correlated with confocal fluorescence microscopy.

Evaluación del tratamiento de linfangiomas con bleomicina en población pediátrica de 0 a 18 años / Evaluation of the treatment of lymphangiomas with bleomycin in a pediatric population aged 0 to 18 years

INTRODUCCIÓN. Los linfangiomas son una malformación infrecuente a escala mundial y constituye una preocupación para los padres del infante; está asociado a problemas estéticos y a posibles efectos deletéreos debido a la obstrucción o compresión de órganos vitales. Se ha descrito a la escleroterapia como la mejor opción de tratamiento. OBJETIVO. Determinar la eficiencia del tratamiento con bleomicina en linfangiomas en la población pediátrica de 0 a 18 años. MATERIALES Y MÉTODOS. Estudio transversal analítico retrospectivo. Población y muestra conocida de 20 datos de Historias Clínicas electrónicas de pacientes diagnosticados con linfangiomas y tratados con bleomicina en el Hospital de Especialidades Carlos Andrade Marín, desde enero 2015 a enero 2018. Criterios de inclusión: pacientes de 0 a 18 años de edad con diagnóstico de linfangioma mediante ecografía y angiotomografía computarizada. Criterios de exclusión: pacientes mayores de 18 años de edad o sospecha diagnóstica de linfangioma sin estudios de imagen, y/o que no acudieron a la cita de control, pacientes diagnosticados de linfangioma que no recibieron bleomicina para su tratamiento, niños con otras malformaciones vasculares. El análisis de datos se realizó en el programa estadístico International Business Machines Statistical Package for the Social Sciences. RESULTADOS. La mediana de edad en mujeres fue de 6,25 años y 3,8 años en hombres. 10 pacientes fueron hombres. El promedio de seguimiento fue de 26,86 +/- 16,78 meses. El tamaño promedio de los linfangiomas fue de 5,77 +/- 3,73 cm. La localización más frecuente fue cervical con un 52,38%. La mayoría fueron macronodulares con un 85,71%. La respuesta fue buena o excelente en el 81,00% de los casos con la primera infiltración y subió al 95,00% con la segunda y tercera, según requerimiento. CONCLUSIÓN. El tratamiento de los linfangiomas con bleomicina fue muy efectivo en la población estudiada.

INTRODUCTION. Lymphatic malformation is a rare malformation worldwide and is a concern for the parents of the infant; it is associated with aesthetic problems and possible deleterious effects due to obstruction or compression of vital organs. Sclerotherapy has been described as the best treatment option. OBJECTIVE. To determine the efficiency of bleomycin treatment in lymphangiomas in the pediatric population aged 0 to 18 years. MATERIALS AND METHODS. Retrospective analytical cross-sectional study. Population and known sample of 20 data from Electronic Medical Records of patients diagnosed with lymphangiomas and treated with bleomycin at the Carlos Andrade Marín Specialties Hospital, from January 2015 to January 2018. Inclusion criteria: patients aged 0 to 18 years with diagnosis of lymphangioma by ultrasound and computed angiotomography. Exclusion criteria: patients older than 18 years of age or diagnostic suspicion of lymphangioma without imaging studies, and/or who did not attend the control appointment, patients diagnosed with lymphangioma who did not receive bleomycin for treatment, children with other vascular malformations. Data analysis was performed in the statistical program International Business Machines Statistical Package for the Social Sciences. RESULTS. The median age in women was 6,25 years and 3,8 years in men. Ten patients were men. The average follow-up was 26,86 +/- 16,78 months. The average size of the lymphatic malformations was 5,77 +/- 3,73 cm. The most frequent location was cervical with 52,38%. Most were macronodular with 85,71%. The response was good or excellent in 81,00% of cases with the first infiltration and rose to 95,00% with the second and third, as required. CONCLUSION. The treatment of lymphangiomas with bleomycin was very effective in the population studied.

Enriquecimiento del aceite comestible por compuestos fenólicos y antioxidantes de chile piquín (Capsicum annuum var. glabriusculum) / Enrichment of edible oil by phenolic and antioxidant compounds of piquín chili (Capsicum annuum var. glabriusculum)

Resumen Capsicum annuum var. glabriusculum presenta adaptaciones locales a distintas dinámicas antropocéntricas y ecológicas, por lo que ampliar sus usos alimenticios a partir de su potencial antioxidante permitirá contribuir al conocimiento para fortalecer cadenas de valor, robustecer su aprovechamiento y el consumo de plantas comestibles silvestres locales. El objetivo de este trabajo fue suplementar con chile piquín un aceite comestible comercial de cártamo y evaluar su capacidad antioxidante, a través de la determinación del contenido de compuestos fenólicos y mediante ensayos de captación de radicales libres ABTS+ y DPPH·, evaluados en: 1) la muestra de chile piquín a través de dos solventes de extracción y 2) en el aceite suplementado y sin suplementar a los 0 d, 7 d, 14 d, 21 d y 28 d posteriores a la preparación (DPP). El chile piquín presentó altos valores antioxidantes, el análisis de varianza (ANOVA) indicó que el extracto acuoso superó al hidroalcohólico. El ANOVA factorial mostró diferencias significativas en los tres parámetros antioxidantes evaluados. Estas disimilitudes se asociaron a la suplementación, tiempo (DPP) y la combinación de la suplementación y DPP. La suplementación del aceite de cártamo con C. annuum var. glabriusculum enriqueció 66 % su valor antioxidante. La comparación de medias mostró diferencias significativas en la combinación de tratamientos y DPP. La combinación de aceite suplementado y los DPP presentó variabilidad en los datos de polifenoles totales y habilidad contra ABTS+. Se observó una relación inversa entre los DPP y compuestos fenólicos totales y capacidad contra ABTS+, contrario a la prueba para DPPH·. Los resultados obtenidos validan que la adición de chile piquín incrementa la biofuncionalidad del aceite de cártamo y puede ser alternativa de fuente de antioxidantes naturales.

Abstract Capsicum annuum var. glabriusculum presents local adaptations to different anthropocentric and ecological dynamics. Therefore, expanding its food uses based on its antioxidant potential will contribute to knowledge about ways to strengthen value chains, enhance their use and encourage the consumption of local wild edible plants. The aim of this work was to evaluate the antioxidant capacity of commercial edible safflower oil supplemented with piquín chili by determining the content of phenolic compounds and by assays of free radical scavenging in ABTS+ and DPPH·. The evaluation included: 1) the piquín chili sample through two extraction solvents and 2) the supplemented and unsupplemented oil at 0 d,7 d, 14 d, 21 d and 28 d after preparation (DPP). The piquín chili presented high antioxidant values. The analysis of variance (ANOVA) indicated that the aqueous extract surpassed the hydroalcoholic extract. The factorial ANOVA showed significant differences in the three antioxidant parameters evaluated. These dissimilarities were associated with supplementation, time (DPP) and the combination of supplementation and DPP. Safflower oil supplementation with C. annuum var. glabriusculum enriched 66 % more its antioxidant value. The comparison of means showed significant differences in the combination of treatments and DPP. The combination of supplemented oil and DPP showed variability in total polyphenol data and ability against ABTS +. An inverse relationship was observed between DPP and total phenolic compounds, and capacity against ABTS +, contrary to the test for DPPH·. The results obtained validate the argument that the addition of piquín chili increases the biofunctionality of safflower oil and can be an alternative source of natural antioxidants.

A Point-of-Care Based on Label-Free Interferometric Optical Detection Method to Evaluate Interferon Gamma (IFN-γ): A Correlation with the ELISA Technique.

Interferon-gamma (IFN-γ) is a cytokine associated with inflammatory diseases, virus, infection, etc. The quantification of interferon-gamma concentration levels is studied to relate the immune system response to the progression of disease. In this work, we used a label-free point-of-care device based on the increase relative optical power (IROP) and a biosensor based on photonic transducers called BICELLs (Biophotonic Sensing Cells) to evaluate interferon-gamma concentrations. The BICELLs' sensing surface size used is 100 µm in diameter. The bioreceptor is attached to the surface by streptavidin-biotin affinity. This label-free IROP-based device can work with a low concentration of reagents and a low sample volume for measurements. Furthermore, this new device was compared with an ELISA technique in the same conditions. A good correlation was achieved between both techniques. This device is easy to use, and it is a cost-effective tool for monitoring an analyte in a disease.

Variability in the Phytochemical Contents and Free Radical-Scavenging Capacity of Capsicum annuum var. glabriusculum (Wild Piquin Chili).

The variability in the phytochemical concentrations of Capsicum annuum var. glabriusculum has not been extensively analyzed among wild populations and ecologic niches in its phylogeographic area. This study aimed to determine the variations in the phytochemical and antioxidant contents of the wild Piquin chili. The total flavonoid content of hydroalcoholic extracts (0.06 to 0.70 mg equivalent of quercetin per gram of dry weight (mg QE/g DW)), free radical-scavenging capacity for 2,2-diphenyl-1-picrylhydrazyl (DPPH. ) radicals (0.55 to 8.55 mm TE/g DW), amount of 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+ ) in aqueous extracts (18.13 to 107.6 mm TE/g DW) and pungency (21,760 to 88,476 Scoville heat units) were highly variable. By analyzing the spatial distribution using the first three principal components, correlations between the phytochemical content and the free radical-scavenging capacity (in both extracts) and flavonoid and phenolic contents (in the hydroalcoholic extract) were observed. Consistent with the statistical analysis, the spatial analysis showed intraregional differences in composition patterns, with an emphasis on central and coastal areas. Flavonoid contents, polyphenol contents and free radical-scavenging activity were the phytochemical components that mainly contributed to the diversity of the population.

Drying Effects on Phenolics and Free Radical-Scavenging Capacity of Rhus pachyrrhachis and Rhus virens Used in Traditional Medicine.

Rhus pachyrrhachis and Rhus virens are medicinal plant species with important uses in northeastern Mexico. They belong to a complex of Rhus species called "lantriscos", which are used for medicinal applications. The medicinal effects of these species are based on traditional use, however, they require phytochemical research to validate their medicinal properties, as well as structural characterization for their correct identification during the collecting practice and uses. The phytochemical potential of aqueous extracts from R. pachyrrhachis and R. virens was analyzed by the quantification of total phenolic content (TPC), free radical-scavenging potential, and total flavonoids, with a comparison of four drying methods, and some phenolic compounds were identified. Furthermore, the stems and leaves of both species were anatomically characterized to establish a differentiation. R. pachyrrhachis and R. virens showed similar values of phytochemical contents, although the TPC content (0.17 mg of gallic acid equivalent per gram of dry weight, GAE/g DW) was higher in R. virens. The drying method used affected the metabolite contents, and this behavior was related to the species. Regarding the phenolic compounds, shikimic acid, galloylquinic acid, and gallic acid were identified in both species, however, quinic acid was only found in Rhus pachyrrhachis, while vanillic acid O-hexoside was identified only in Rhus virens. At the anatomical level, the pubescence associated with trichomes on the leaves of Rhus pachyrrhachis was highlighted as the main differential characteristic.

Social and environmental factors in the diversity of tomato landraces from the South-Central region of Mexico / Influência de fatores sociais e ambientais na diversidade de variedades crioulas de tomateiro da região Centro-Sul do México

ABSTRACT: In the present study, we investigated the influence of social and environmental factors in the genetic diversity of tomato landraces in the South-Central region of Mexico. A total of 30 tomato landraces, collected in 18 villages with different ethnolinguistic affiliations, were analyzed. We reported that the genetic diversity of tomato landraces is associated with the ethnolinguistic group, weather, and soil-type present in the region studied. Our results showed that there are morphological differences between landraces grown by different ethnolinguistic groups; however, there was also evidence of morphological similarities between landraces from groups with different ethnolinguistic affiliations. Finally, different selection criteria, mainly fruit color, size and shape, plays an important role in the phenotypic divergence among landraces grown in different traditional farming systems.

RESUMO: No presente estudo investigamos a relação da afiliação etnolinguística com a diversidade genética de variedades locais de tomate na região Centro-Sul do México. Um total de 30 variedades de tomate, coletadas em 18 aldeias com diferentes afiliações etnolinguísticas, foram analisadas. Verificamos que a diversidade genética das variedades crioulas de tomate está associada ao grupo etnolinguístico presente na região estudada. Nossos resultados mostram que existem diferenças morfológicas entre raças cultivadas por diferentes grupos etnolinguísticos. No entanto, também houve evidências de semelhanças morfológicas entre raças de grupos com diferentes afiliações etnolinguísticas. Finalmente, diferentes critérios de seleção, como, principalmente, cor, tamanho e forma dos frutos, desempenham um papel importante na divergência fenotípica entre as variedades crioulas cultivadas em diferentes sistemas agrícolas tradicionais.

WadD, a New Brucella Lipopolysaccharide Core Glycosyltransferase Identified by Genomic Search and Phenotypic Characterization.

Brucellosis, an infectious disease caused by Brucella, is one of the most extended bacterial zoonosis in the world and an important cause of economic losses and human suffering. The lipopolysaccharide (LPS) of Brucella plays a major role in virulence as it impairs normal recognition by the innate immune system and delays the immune response. The LPS core is a branched structure involved in resistance to complement and polycationic peptides, and mutants in glycosyltransferases required for the synthesis of the lateral branch not linked to the O-polysaccharide (O-PS) are attenuated and have been proposed as vaccine candidates. For this reason, the complete understanding of the genes involved in the synthesis of this LPS section is of particular interest. The chemical structure of the Brucella LPS core suggests that, in addition to the already identified WadB and WadC glycosyltransferases, others could be implicated in the synthesis of this lateral branch. To clarify this point, we identified and constructed mutants in 11 ORFs encoding putative glycosyltransferases in B. abortus. Four of these ORFs, regulated by the virulence regulator MucR (involved in LPS synthesis) or the BvrR/BvrS system (implicated in the synthesis of surface components), were not required for the synthesis of a complete LPS neither for virulence or interaction with polycationic peptides and/or complement. Among the other seven ORFs, six seemed not to be required for the synthesis of the core LPS since the corresponding mutants kept the O-PS and reacted as the wild type with polyclonal sera. Interestingly, mutant in ORF BAB1_0953 (renamed wadD) lost reactivity against antibodies that recognize the core section while kept the O-PS. This suggests that WadD is a new glycosyltransferase adding one or more sugars to the core lateral branch. WadD mutants were more sensitive than the parental strain to components of the innate immune system and played a role in chronic stages of infection. These results corroborate and extend previous work indicating that the Brucella LPS core is a branched structure that constitutes a steric impairment preventing the elements of the innate immune system to fight against Brucella.

Free Radical-Scavenging Capacities, Phenolics and Capsaicinoids in Wild Piquin Chili (Capsicum annuum var. Glabriusculum).

The total phenolic compounds content, free radical-scavenging capacity and capsaicinoid content in populations of wild Piquin chili (C. annuum) were studied. Aqueous and hydroalcoholic extracts from nine ecotypes were evaluated. High contents of phenolic compounds and free radical-scavenging capacities were observed for both extracts; however, the values that were found for the hydroalcoholic phase were substantially higher. LC-MS analysis allowed for the detection of 32 compounds, where apigenin-8-C-glucoside followed by vanillic acid 1-O-ß-o-glucopyranosylester (Isomer I or II) and 7-ethoxy-4-methylcoumarin were the most widely distributed; they were found in more than 89% of the ecotypes. The diversity of identified phenolic compounds was different among ecotypes, allowing them to be distinguished by chemical diversity, free radical-scavenging capacities and heat Scoville units. The total capsaicinoid content was higher in Population I (23.5 mg/g DW) than in Populations II and III, which had contents of 15.3 and 10.7 mg/g DW, respectively. This variability could lead to phytochemical exploitation and the conservation of the natural populations of wild chili.

Genomic Insertion of a Heterologous Acetyltransferase Generates a New Lipopolysaccharide Antigenic Structure in Brucella abortus and Brucella melitensis.

Brucellosis is a bacterial zoonosis of worldwide distribution caused by bacteria of the genus Brucella. In Brucella abortus and Brucella melitensis, the major species infecting domestic ruminants, the smooth lipopolysaccharide (S-LPS) is a virulence factor. This S-LPS carries a N-formyl-perosamine homopolymer O-polysaccharide that is the major antigen in serodiagnostic tests and is required for virulence. We report that the Brucella O-PS can be structurally and antigenically modified using wbdR, the acetyl-transferase gene involved in N-acetyl-perosamine synthesis in Escherichia coli O157:H7. Brucella constructs carrying plasmidic wbdR expressed a modified O-polysaccharide but were unstable, a problem circumvented by inserting wbdR into a neutral site of chromosome II. As compared to wild-type bacteria, both kinds of wbdR constructs expressed shorter O-polysaccharides and NMR analyses showed that they contained both N-formyl and N-acetyl-perosamine. Moreover, deletion of the Brucella formyltransferase gene wbkC in wbdR constructs generated bacteria producing only N-acetyl-perosamine homopolymers, proving that wbdR can replace for wbkC. Absorption experiments with immune sera revealed that the wbdR constructs triggered antibodies to new immunogenic epitope(s) and the use of monoclonal antibodies proved that B. abortus and B. melitensis wbdR constructs respectively lacked the A or M epitopes, and the absence of the C epitope in both backgrounds. The wbdR constructs showed resistance to polycations similar to that of the wild-type strains but displayed increased sensitivity to normal serum similar to that of a per R mutant. In mice, the wbdR constructs produced chronic infections and triggered antibody responses that can be differentiated from those evoked by the wild-type strain in S-LPS ELISAs. These results open the possibilities of developing brucellosis vaccines that are both antigenically tagged and lack the diagnostic epitopes of virulent field strains, thereby solving the diagnostic interference created by current vaccines against Brucella.

On the Determination of Uncertainty and Limit of Detection in Label-Free Biosensors.

A significant amount of noteworthy articles reviewing different label-free biosensors are being published in the last years. Most of the times, the comparison among the different biosensors is limited by the procedure used of calculating the limit of detection and the measurement uncertainty. This article clarifies and establishes a simple procedure to determine the calibration function and the uncertainty of the concentration measured at any point of the measuring interval of a generic label-free biosensor. The value of the limit of detection arises naturally from this model as the limit at which uncertainty tends when the concentration tends to zero. The need to provide additional information, such as the measurement interval and its linearity, among others, on the analytical systems and biosensor in addition to the detection limit is pointed out. Finally, the model is applied to curves that are typically obtained in immunoassays and a discussion is made on the application validity of the model and its limitations.

Identification of lptA, lpxE, and lpxO, Three Genes Involved in the Remodeling of Brucella Cell Envelope.

The brucellae are facultative intracellular bacteria that cause a worldwide extended zoonosis. One of the pathogenicity mechanisms of these bacteria is their ability to avoid rapid recognition by innate immunity because of a reduction of the pathogen-associated molecular pattern (PAMP) of the lipopolysaccharide (LPS), free-lipids, and other envelope molecules. We investigated the Brucella homologs of lptA, lpxE, and lpxO, three genes that in some pathogens encode enzymes that mask the LPS PAMP by upsetting the core-lipid A charge/hydrophobic balance. Brucella lptA, which encodes a putative ethanolamine transferase, carries a frame-shift in B. abortus but not in other Brucella spp. and phylogenetic neighbors like the opportunistic pathogen Ochrobactrum anthropi. Consistent with the genomic evidence, a B. melitensis lptA mutant lacked lipid A-linked ethanolamine and displayed increased sensitivity to polymyxin B (a surrogate of innate immunity bactericidal peptides), while B. abortus carrying B. melitensis lptA displayed increased resistance. Brucella lpxE encodes a putative phosphatase acting on lipid A or on a free-lipid that is highly conserved in all brucellae and O. anthropi. Although we found no evidence of lipid A dephosphorylation, a B. abortus lpxE mutant showed increased polymyxin B sensitivity, suggesting the existence of a hitherto unidentified free-lipid involved in bactericidal peptide resistance. Gene lpxO putatively encoding an acyl hydroxylase carries a frame-shift in all brucellae except B. microti and is intact in O. anthropi. Free-lipid analysis revealed that lpxO corresponded to olsC, the gene coding for the ornithine lipid (OL) acyl hydroxylase active in O. anthropi and B. microti, while B. abortus carrying the olsC of O. anthropi and B. microti synthesized hydroxylated OLs. Interestingly, mutants in lptA, lpxE, or olsC were not attenuated in dendritic cells or mice. This lack of an obvious effect on virulence together with the presence of the intact homolog genes in O. anthropi and B. microti but not in other brucellae suggests that LptA, LpxE, or OL ß-hydroxylase do not significantly alter the PAMP properties of Brucella LPS and free-lipids and are therefore not positively selected during the adaptation to intracellular life.