Multi-species biofilms of environmental microbiota isolated from fruit packing facilities promoted tolerance of Listeria monocytogenes to benzalkonium chloride.

Listeria monocytogenes may survive and persist in food processing environments due to formation of complex multi-species biofilms of environmental microbiota that co-exists in these environments. This study aimed to determine the effect of selected environmental microbiota on biofilm formation and tolerance of L. monocytogenes to benzalkonium chloride in formed biofilms. The studied microbiota included bacterial families previously shown to co-occur with L. monocytogenes in tree fruit packing facilities, including Pseudomonadaceae, Xanthomonadaceae, Microbacteriaceae, and Flavobacteriaceae. Biofilm formation ability and the effect of formed biofilms on the tolerance of L. monocytogenes to benzalkonium chloride was measured in single- and multi-family assemblages. Biofilms were grown statically on polystyrene pegs submerged in a R2A broth. Biofilm formation was quantified using a crystal violet assay, spread-plating, confocal laser scanning microscopy, and its composition was assessed using amplicon sequencing. The concentration of L. monocytogenes in biofilms was determined using the most probable number method. Biofilms were exposed to the sanitizer benzalkonium chloride, and the death kinetics of L. monocytogenes were quantified using a most probable number method. A total of 8, 8, 6, and 3 strains of Pseudomonadaceae, Xanthomonadaceae, Microbacteriaceae, and Flavobacteriaceae, respectively, were isolated from the environmental microbiota of tree fruit packing facilities and were used in this study. Biofilms formed by Pseudomonadaceae, Xanthomonadaceae, and all multi-family assemblages had significantly higher concentration of bacteria, as well as L. monocytogenes, compared to biofilms formed by L. monocytogenes alone. Furthermore, multi-family assemblage biofilms increased the tolerance of L. monocytogenes to benzalkonium chloride compared to L. monocytogenes mono-species biofilms and planktonic multi-family assemblages. These findings suggest that L. monocytogenes control strategies should focus not only on assessing the efficacy of sanitizers against L. monocytogenes, but also against biofilm-forming microorganisms that reside in the food processing built environment, such as Pseudomonadaceae or Xanthomonadaceae.

Sourdough starter culture microbiomes influence physical and chemical properties of wheat bread.

Sourdough fermentation is an ancient leavening method that uses wild yeasts to produce carbon dioxide, contributing to bread rise, and bacteria which produce organic acids. Sourdough starter cultures are known to be diverse in terms of the microorganisms they comprise and while specific genera and species of microorganisms have been identified from starters and associated with specific attributes, overarching relationships between sourdough starter culture microbiomes and bread quality are not well understood. The objective of this study was to characterize differences in the physical and chemical properties of breads produced with sourdough starter cultures with unique microbiomes. Sourdough starter cultures (n = 20) of known microbial populations were used to produce wheat-based dough and bread, which were analyzed for chemical and physical properties then compared to their microbial populations in order to identify relationships between microbial profiles and dough/bread qualities. All samples were also compared to bread produced only with Saccharomyces cerevisiae (baker's yeast). Significant differences among pH, titratable acidity, loaf volume, crumb firmness, crust color, free amino acids, and organic acids were observed when comparing sourdough breads to the yeast-only control (p ≤ 0.05). Furthermore, bacterial diversity of sourdough starter cultures was correlated with lactic acid and free amino acid in the dough and loaf volume and crumb firmness of baked breads. No significant correlations were found between fungal diversity and measured outcomes. These data demonstrate the importance of considering sourdough starter microbiomes as an ingredient in baked goods and they contribute to quality and safety outcomes in bread production. PRACTICAL APPLICATION: Sourdough starter cultures have diverse and dynamic populations of bacteria and yeasts, which contribute to the production of bread products. These populations can influence the physical and chemical properties of sourdough fermentation and final breads. Understanding of the relationship between sourdough starter microbiomes and bread quality parameters can lead to targeted development of sourdough bread products with specific physical and chemical properties.

Context matters: environmental microbiota from ice cream processing facilities affected the inhibitory performance of two lactic acid bacteria strains against Listeria monocytogenes.

IMPORTANCE: Antilisterial LAB strains have been proposed as biological control agents for application in food processing environments. However, the effect of resident food processing environment microbiota on the performance on antilisterial LAB strains is poorly understood. Our study shows that the presence of microbiota collected from ice cream processing facilities' environmental surfaces can affect the attachment and inhibitory effect of LAB strains against L. monocytogenes. Further studies are therefore needed to assess whether individual microbial taxa affect antilisterial properties of LAB strains and to characterize the underlying mechanisms.

The composition of environmental microbiota in three tree fruit packing facilities changed over seasons and contained taxa indicative of L. monocytogenes contamination.

BACKGROUND: Listeria monocytogenes can survive in cold and wet environments, such as tree fruit packing facilities and it has been implicated in outbreaks and recalls of tree fruit products. However, little is known about microbiota that co-occurs with L. monocytogenes and its stability over seasons in tree fruit packing environments. In this 2-year longitudinal study, we aimed to characterize spatial and seasonal changes in microbiota composition and identify taxa indicative of L. monocytogenes contamination in wet processing areas of three tree fruit packing facilities (F1, F2, F3). METHODS: A total of 189 samples were collected during two apple packing seasons from floors under the washing, drying, and waxing areas. The presence of L. monocytogenes was determined using a standard culturing method, and environmental microbiota was characterized using amplicon sequencing. PERMANOVA was used to compare microbiota composition among facilities over two seasons, and abundance-occupancy analysis was used to identify shared and temporal core microbiota. Differential abundance analysis and random forest were applied to detect taxa indicative of L. monocytogenes contamination. Lastly, three L. monocytogenes-positive samples were sequenced using shotgun metagenomics with Nanopore MinION, as a proof-of-concept for direct detection of L. monocytogenes' DNA in environmental samples. RESULTS: The occurrence of L. monocytogenes significantly increased from 28% in year 1 to 46% in year 2 in F1, and from 41% in year 1 to 92% in year 2 in F3, while all samples collected from F2 were L. monocytogenes-positive in both years. Samples collected from three facilities had a significantly different microbiota composition in both years, but the composition of each facility changed over years. A subset of bacterial taxa including Pseudomonas, Stenotrophomonas, and Microbacterium, and fungal taxa, including Yarrowia, Kurtzmaniella, Cystobasidium, Paraphoma, and Cutaneotrichosporon, were identified as potential indicators of L. monocytogenes within the monitored environments. Lastly, the DNA of L. monocytogenes was detected through direct Nanopore sequencing of metagenomic DNA extracted from environmental samples. CONCLUSIONS: This study demonstrated that a cross-sectional sampling strategy may not accurately reflect the representative microbiota of food processing facilities. Our findings also suggest that specific microorganisms are indicative of L. monocytogenes, warranting further investigation of their role in the survival and persistence of L. monocytogenes. Video Abstract.

Ability of Two Strains of Lactic Acid Bacteria To Inhibit Listeria monocytogenes by Spot Inoculation and in an Environmental Microbiome Context.

We evaluated the ability of two strains of lactic acid bacteria (LAB) to inhibit L. monocytogenes using spot inoculation and environmental microbiome attached-biomass assays. LAB strains (PS01155 and PS01156) were tested for antilisterial activity toward 22 phylogenetically distinct L. monocytogenes strains isolated from three fruit packing environments (F1, F2, and F3). LAB strains were tested by spot inoculation onto L. monocytogenes lawns (108 and 107 CFU/mL) and incubated at 15, 20, 25, or 30°C for 3 days. The same LAB strains were also cocultured at 15°C for 3, 5, and 15 days in polypropylene conical tubes with L. monocytogenes and environmental microbiome suspensions collected from F1, F2, and F3. In the spot inoculation assay, PS01156 was significantly more inhibitory toward less concentrated L. monocytogenes lawns than more concentrated lawns at all the tested temperatures, while PS01155 was significantly more inhibitory toward less concentrated lawns only at 15 and 25°C. Furthermore, inhibition of L. monocytogenes by PS01156 was significantly greater at 15°C than higher temperatures, whereas the temperature did not have an effect on the inhibitory activity of PS01155. In the assay using attached environmental microbiome biomass, L. monocytogenes concentration was significantly reduced by PS01156, but not PS01155, when cocultured with microbiomes from F1 and F3 and incubated for 3 days at 15°C. Attached biomass microbiota composition was significantly affected by incubation time but not by LAB strain. This study demonstrates that LAB strains that may exhibit inhibitory properties toward L. monocytogenes in a spot inoculation assay may not maintain antilisterial activity within a complex microbiome. IMPORTANCE Listeria monocytogenes has previously been associated with outbreaks of foodborne illness linked to consumption of fresh produce. In addition to conventional cleaning and sanitizing, lactic acid bacteria (LAB) have been studied for biocontrol of L. monocytogenes in food processing environments that are challenging to clean and sanitize. We evaluated whether two specific LAB strains, PS01155 and PS01156, can inhibit the growth of L. monocytogenes strains in a spot inoculation and in an attached-biomass assay, in which they were cocultured with environmental microbiomes collected from tree fruit packing facilities. LAB strains PS01155 and PS01156 inhibited L. monocytogenes in a spot inoculation assay, but the antilisterial activity was lower or not detected when they were grown with environmental microbiota. These results highlight the importance of conducting biocontrol challenge tests in the context of the complex environmental microbiomes present in food processing facilities to assess their potential for application in the food industry.

Matrix Effects on the Delivery Efficacy of Bifidobacterium animalis subsp. lactis BB-12 on Fecal Microbiota, Gut Transit Time, and Short-Chain Fatty Acids in Healthy Young Adults.

Probiotics are consumed in fermented dairy products or as capsules for their putative health benefits. However, little research has been done to evaluate the effects of the delivery matrix on the health benefits of probiotics in humans. To examine the effects of delivering Bifidobacterium animalis subsp. lactis BB-12 (BB-12) (log10 10 ± 0.5 CFU/day) via a yogurt smoothie versus a capsule, we monitored the fecal microbiota, gut transit times (GTTs), and fecal excretion of short-chain fatty acids (SCFAs) in healthy adults. In a randomized, four-period, crossover study performed in a partially blind manner, 36 adults were recruited and randomly assigned to four treatments: control yogurt smoothie (YS), yogurt smoothie with BB-12 added prefermentation (PRE), yogurt smoothie with BB-12 added postfermentation (POST), and capsule containing BB-12 (CAP). Participants' fecal microbiota was assessed using 16S rRNA sequencing, GTTs via SmartPill, and fecal SCFAs by gas chromatography (GC) before (baseline) and after each intervention. Participants had significantly higher percentage of Streptococcus after consuming YS versus CAP (P = 0.01). Bifidobacterium-specific terminal restriction fragment length polymorphism analysis revealed a significantly higher percentage of B. animalis after consuming PRE and POST compared to baseline, YS, CAP, and final washout (P < 0.0001). The predominant SCFAs were negatively correlated with GTTs. Consumption of BB-12 delivered in a yogurt smoothie or capsule did not significantly alter the composition of the gut microbiota, GTTs, or fecal SCFA concentration of the study cohort. However, daily consumption of BB-12 in yogurt smoothie may result in higher relative abundance of B. animalis in healthy adults. (This trial has been registered at ClinicalTrials.gov under identifier NCT01399996.) IMPORTANCE Bifidobacterium animalis subsp. lactis BB-12 is a probiotic strain that has been used worldwide since 1985. It has commonly been delivered in fermented dairy products for perceived benefits associated with gut health and enhanced immune function. In addition to fermented dairy products, many new probiotic-containing alternatives such as probiotic-containing juice, probiotic-containing chocolate, and capsules have been developed. While these products provide more options for people to access probiotics, little research has been done on the effect of delivery matrix (dairy versus nondairy) on their efficacy in humans. In addition, it was unclear how yogurt fermentation may influence the survival of BB-12 in the product or on its performance in vivo. The significance of our study is in simultaneously assessing the effect of BB-12, alone and in different delivery vehicles, on the gut transit time, fecal short-chain fatty acids, and the composition of the gut microbiota of the study cohort.

Review of the prevalence of foodborne pathogens in milk and dairy products in Ethiopia.

Food safety is a significant barrier to social and economic development throughout the world, particularly in developing countries. Here, we reviewed the prevalence of major bacterial foodborne pathogens (Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7 and Campylobacter spp.) in the rapidly growing Ethiopian dairy supply-chain. We identified 15, 9, 5 and 0 studies that had reported the prevalence of Salmonella spp., L. monocytogenes, E. coli O157:H7, and Campylobacter spp. in dairy foods, respectively. The studies reviewed reported a median prevalence of Salmonella, L. monocytogenes, and E. coli O157:H7 of 6, 9 and 10%, respectively, in raw cow milk in Ethiopia, indicating a concerning occurrence of bacterial foodborne pathogens in raw milk. Implementation of good hygiene and production practices and assessment of interventions targeting the reduction of contamination in the dairy supply chain is needed to inform coordinated efforts focused on improvement of dairy food safety in Ethiopia.

Intervención temprana tras los terremotos de Puerto Rico en enero de 2020: resiliencia, prevención y promoción de la salud mental infantojuvenil / Early Intervention After the Earthquakes in Puerto Rico in January 2020: Resilience, Prevention and Promotion of Child and Adolescent Mental Health

INTRODUCCIÓN: El artículo describe un modelo de intervención temprana para la prevención de patología, y promoción de la resiliencia, tras los terremotos de la isla de Puerto Rico en enero de 2020. El 7 de enero de 2020 un terremoto de magnitud 6,4 en la escala Richter, sacudió la isla de Puerto Rico. Como consecuencia, gran parte de la población sufrió desplazamientos de sus domicilios y cierres de muchas escuelas. Todo ello sobre lo ya anteriormente vivido, como el Huracán María en 2017, aumentando así su carga de estrés alostática. Tras la ayuda inicial de los cuerpos de rescate, se detecta la necesidad de apoyo psicológico a la población y a profesionales proveedores de dicho apoyo. MÉTODOS: Contando con una precaria infraestructura, se decide realizar una intervención con el objetivo de proporcionar información del impacto del estrés traumático en el organismo, así como ejercicios prácticos de mindfulness dirigidos a dichos profesionales. El programa integra lecciones de la Terapia de Claves Traumáticas y ejercicios del currículo Pure Power. RESULTADOS: Se confirma, a través de un cuestionario de satisfacción, la necesidad de este tipo de programas, y se valora extrapolar el modelo a otras poblaciones de riesgo similares, en términos de evitar las consecuencias del trauma, disminuir la carga alostática, y prevenir y promocionar la salud mental. Se sugiere realizar próximas formaciones de ampliación de este programa, para confirmar su efectividad y mantenimiento en el tiempo

INTRODUCTION: This article describes an early intervention model for the prevention of pathology, and for the promotion of resilience, after the earthquakes on the island of Puerto Rico in January 2020. On January 7, 2020, an earthquake of magnitude 6.4 on the Richter scale struck the island of Puerto Rico. As a consequence of the seismic movement, an important part of the population, suffered displacements from their homes and closing of schools. In addition to what they previously lived through, such as Hurricane Maria in 2017, increasing their burden of allostatic stress. After initial help from the rescue forces, a need to provide psychological support to the population and to the professionals involved in giving that psychological support, was detected. METHODS: Starting from a precarious infrastructure, it was decided to implement an intervention with the objective of providing information on the impact of traumatic stress on the individual, along with practical mindfulness exercises. The program integrated lessons from Cue-Centered Therapy with exercises from the Pure Power curriculum. RESULTS: Through a satisfaction questionnaire, the need of these type of programs was confirmed. The implementation of these prevention programs should be considered when planning to intervene with other similar populations at risk. Thus, avoiding the consequences of trauma, reducing the allostatic load in the individual, and preventing and promoting mental health and resilience. The discussion introduces the need to develop follow-up procedures, to confirm the effectiveness and maintenance over time

Psoriasis linear y liquen estriado: una asociación rara entre dos dermatosis lineares / Linear psoriasis and lichen striatus: a rare association between two linear dermatoses

Resumen: La psoriasis linear y el liquen estriado son dos dermatosis de distribución linear por su localización sobre las líneas de Blaschko. Aunque estas dos enfermedades comparten algunas características, su asociación no es común y su diagnóstico en algunas ocasiones puede ser un reto. Presentamos el caso de una paciente pediátrica que desarrolló las dos patologías, con adecuada respuesta al tratamiento con corticoides tópicos.

Abstract: Linear Psoriasis and Lichen Striatus are known as linear dermatoses for their distribution along the Blaschko lines. Although they share some characteristics, their association is not common and accurate diagnosis can sometimes be a challenge. We present the case of a girl who develops two linear dermatoses, with adequate response to topical corticosteroids.

Effect of fat content on the physical properties and consumer acceptability of vanilla ice cream.

Ice cream is a complex food matrix that contains multiple physical phases. Removal of 1 ingredient may affect not only its physical properties but also multiple sensory characteristics that may or may not be important to consumers. Fat not only contributes to texture, mouth feel, and flavor, but also serves as a structural element. We evaluated the effect of replacing fat with maltodextrin (MD) on select physical properties of ice cream and on consumer acceptability. Vanilla ice creams were formulated to contain 6, 8, 10, 12, and 14% fat, and the difference was made up with 8, 6, 4, 2, and 0% maltodextrin, respectively, to balance the mix. Physical characterization included measurements of overrun, apparent viscosity, fat particle size, fat destabilization, hardness, and melting rate. A series of sensory tests were conducted to measure liking and the intensity of various attributes. Tests were also conducted after 19 weeks of storage at -18°C to assess changes in acceptance due to prolonged storage at unfavorable temperatures. Then, discrimination tests were performed to determine which differences in fat content were detectable by consumers. Mix viscosity decreased with increasing fat content and decreasing maltodextrin content. Fat particle size and fat destabilization significantly increased with increasing fat content. However, acceptability did not differ significantly across the samples for fresh or stored ice cream. Following storage, ice creams with 6, 12, and 14% fat did not differ in acceptability compared with fresh ice cream. However, the 8% fat, 6% MD and 10% fat, 4% MD ice creams showed a significant drop in acceptance after storage relative to fresh ice cream at the same fat content. Consumers were unable to detect a difference of 2 percentage points in fat level between 6 and 12% fat. They were able to detect a difference of 4 percentage points for ice creams with 6% versus 10%, but not for those with 8% versus 12% fat. Removing fat and replacing it with maltodextrin caused minimal changes in physical properties in ice cream and mix and did not change consumer acceptability for either fresh or stored ice cream.

Infección por virus del Zika en el embarazo, impacto fetal y neonatal / Zika virus infection in pregnancy, fetal and neonatal impact

Zika virus (ZV) is an arbovirus transmitted by Aedes aegypti and A. albopictus. The neurotropic profile of this virus is known since 1952. The main finding related to ZV in America is microcephaly. Two hypotheses are tested on its involvement in the central nervous system: its neurotropic feature and the direct effect of ZV on the placenta. Malformations and clinical findings on fetal development comprise congenital Zika syndrome. RT-PCR and serology (IgM) are useful for definitive diagnosis. However, we should keep in mind first that the viremia in pregnant women can stay for a longer period of time, and second, a positive IgM for Zika should be properly interpreted in an endemic area to other flavivirus. It is suggested to be part of TORCHS-Z complex the ZV infection in endemic areas.

El virus del Zika (VZ), arbovirus, es transmitido por Aedes aegypti y A. albopictus. Desde 1952 se conoce su perfil neurotrópico. El principal hallazgo relacionado con la infección en las Américas, es la microcefalia. Dos hipótesis se plantean sobre su afectación en el sistema nervioso central: su característica neurotrópica per se, y el efecto directo del virus sobre la placenta. Las malformaciones y hallazgos clínicos sobre el desarrollo fetal conforman el síndrome de Zika congénito. La reacción de polimerasa en cadena-transcriptasa reversa (RPC-TR) y serología (IgM) son útiles para el diagnóstico definitivo; sin embargo, debe tenerse en cuenta, primero, que la viremia en las mujeres embarazadas puede permanecer por un período más prolongado y segundo, que una IgM positiva para Zika, debe ser adecuadamente interpretada en un medio endémico para otros flavivirus. Se propone a la infección por el VZ, en zonas endémicas, como parte del complejo TORCHS-Z.

[Zika virus infection in pregnancy, fetal and neonatal impact]. / Infección por virus del Zika en el embarazo, impacto fetal y neonatal.

Zika virus (ZV) is an arbovirus transmitted by Aedes aegypti and A. albopictus. The neurotropic profile of this virus is known since 1952. The main finding related to ZV in America is microcephaly. Two hypotheses are tested on its involvement in the central nervous system: its neurotropic feature and the direct effect of ZV on the placenta. Malformations and clinical findings on fetal development comprise congenital Zika syndrome. RT-PCR and serology (IgM) are useful for definitive diagnosis. However, we should keep in mind first that the viremia in pregnant women can stay for a longer period of time, and second, a positive IgM for Zika should be properly interpreted in an endemic area to other flavivirus. It is suggested to be part of TORCHS-Z complex the ZV infection in endemic areas.

Comportamientos que favorecen la dinámica de reinfestación de Triatomainfestans del Chaco paraguayo / Behaviors that favor reinfestation by Triatoma infestans in the Paraguayan Chaco

El Chaco central paraguayo es unaregión de alto nivel de reinfestación porTriatoma infestans. La población indígena que la habita tiene alta vulnerabilidad por factores culturales y medio ambientales que dificultan acceso y trabajo en la zona. Se propusoconocer factores psicosociales asociadosal proceso de reinfestación para desarrollar tareas de vigilanciacomunitaria. Estudio transversal, con enfoques cualitativo y cuantitativo.Treinta y seis punto siete por ciento (96)de la población de estudio realizómejoras en viviendas; 41,6 % (40)mejoró revoque en paredes. Poblaciónubica al vector en el monte, entre leñas,hojas secas, agujeros de árboles, pozosde topos o tatú; alrededor de animales domésticos, techos de viviendas, gallineros, chiquero de cabras y cerdos.Comprometiendo el traslado pasivo de vinchucas se encontró la recolección deleña 98,5 % (266), del monte, 97,7 %(261) el cambio de lugar de ropas, cajas y comida en las viviendas, 54,7 % (146). Se asoció (p< 0,0005) vivienda mejorada con revoque en paredes yno infestación; viviendas con animales(p< 0,03) e infestación; actitud positivapara eliminar el vector (p<0,04) y no infestación. Comportamientos que comprometen traslado y permanencia de vinchucas fueron acarreo de leña, almacenamiento de comidas yacumulación de ropa y cajas. Paredes revocadas y presencia de animales domésticos se correlacionaron a infestación y actitudes positivas paraeliminación de la vinchuca con viviendas sin reinfestación. Todos ellos sonfactores estratégicos para tareas deprevención y vigilancia con participación comunitaria.

No reduction of HCV viral load in HIV patients co-infected with HCV genotype 1 during a 30 days course of nitazoxanide monotherapy.

There are two new drugs approved and several in development for treatment of chronic HCV; among them nitazoxanide (NTZ). Twelve HIV/HCV genotype 1 co-infected patients were enrolled prospectively to receive a 30 days course of oral NTZ 500 mg bid. This therapy was well tolerated in this group of HIV patients co-infected with HCV genotype 1. Nevertheless no changes in HCV viral load were observed during treatment in none of the patients evaluated. This data suggests that despite the promising results reported for HCV genotype 4 mono-infected patients, NTZ exhibit poor activity as monotherapy in HIV/HCV co-infected patients with genotype 1.

HCV RNA decline in the first 24 h exhibits high negative predictive value of sustained virologic response in HIV/HCV genotype 1 co-infected patients treated with peginterferon and ribavirin.

BACKGROUND: Treatment with Peg-interferon and ribavirin (PEG-IFN/RBV) for HIV patients co-infected with hepatitis C virus (HCV) genotype 1 has suboptimal rates of response. Viral kinetics has emerged as one of the best prognostic factors of treatment outcome. METHODS: Twenty HIV/HCV genotype 1 co-infected patients in treatment with PEG-IFN/RBV, had blood drawn at baseline, 24 h, 4, 12, 24, 48, and 72 weeks. HCV-RNA levels were evaluated at each time point. ROC curves were used to evaluate the log10 HCV-RNA decay at 24 h that exhibits the best predictive value of achieving response. Genomic characterization of HCV NS5A at both interferon sensitivity-determining region (ISDR) and protein-kinase binding (PKRBD) domains were performed in order to evaluate its heterogeneity and association with 24 h HCV-RNA decay and SVR. RESULTS: Non-responder patients exhibited a mean of 0.7 log10 (SD 0.74 log10) HCV-RNA decay at 24 h, whereas responder-patients presented 1.6 log10 (SD 0.28 log10), p = 0.04. A reduction in HCV viral load from baseline to 24 h of < 1.4 had a negative predictive value for achieving SVR of 100% and a positive predictive value of 50%. HCV genotype 1 isolates from patients with a decrease of HCV-RNA at 24 h > 1.4 log10, exhibited 3.1(SD 1.5) amino acids substitutions in ISDR and 4.8(SD 2.3) in PKRBD regions and 1.6(SD 0.7) and 2.4(SD 1.3), respectively, in those patients presenting lower reduction in HCV-RNA. CONCLUSIONS: HIV/HCV genotype 1 co-infected patients with a decrease in HCV-VL at 24 h > 1.4 log10 are more likely to achieve SVR when treated with PEG-IFN/RBV than those with lower levels of HCV-RNA decay. Along with other host-related and viral-related prognostic factors in HIV/HCV co-infected patients, this very early time point of evaluation could be of relevance in the management of HCV-specific treatment.

A new pharmacological screening assay with Trypanosoma cruzi epimastigotes expressing beta-galactosidase.

We have developed a new pharmacological screening assay for epimastigotes of Trypanosoma cruzi (clone CL-B5) that express the Escherichia coli LacZ gene. The assay is based on determining the activity of the cytoplasmic beta-galactosidase released into the culture on membrane lysis in the presence of the substrate chlorophenol red beta-D-galactopyranoside (CPRG). The experimental conditions were adjusted to find those in which the relationship between epimastigote number and CPRG absorbance was linear over the widest possible range. Absorbance was significantly correlated with the number epimastigote from 5x10(3) to 1.2x10(6) parasites/ml (r=0.98, P<0.01). The optimal final concentration of CPRG was 200 microM and the optimal incubation period was 6 h when parasites were incubated for 3 days. Once the assay was standardized, the trypanocidal activities of nifurtimox and benznidazole were determined both by CPRG assay and microscopic counting, demonstrating the methods utility for drug-screening. The efficacy obtained was comparable to that obtained with the manual method.

Cystic fibrosis-related diabetes mellitus: clinical impact of prediabetes and effects of insulin therapy.

UNLABELLED: In patients with cystic fibrosis (CF), glucose intolerance preceding diabetes (prediabetes) may have adverse effects on nutritional status and respiratory function, which are reversible after the start of insulin therapy. Respiratory function (forced vital capacity and forced expiratory volume in one second) and body mass index (BMI) were compared retrospectively in a French cohort of 14 patients during the 5 y preceding insulin therapy for diabetes and in 14 age- and sex-matched controls with normal oral glucose tolerance tests. In the diabetic group, all three parameters deviated increasingly from the values in the controls; the differences became statistically different during the 6 mo before insulin therapy. The effect was more important in patients for whom diabetes mellitus was diagnosed on the basis of symptoms of hyperglycaemia than in patients for whom it was diagnosed by systematic screening, but still present in the latter. After insulin was started, respiratory function improved and the BMI returned to normal within 1 y. The annual insulin requirement increased from 0.62 during the first year to 1.25 during the fifth year. Glycosylated haemoglobin (HbAIc) values ranged from 6.6 to 7.8%. Only 2 episodes of severe hypoglycaemia were recorded over 42 patient-years of follow-up. The insulin regimen most often used was two daily injections of a mixture of short- and intermediate-acting insulin (n = 10) given with an insulin pen. CONCLUSION: The clinical status of CF patients who will need insulin therapy deteriorates before the start of insulin. In patients with CF-related diabetes, with or without fasting hyperglycaemia, insulin therapy improves anabolism and provides good glycaemic control with few severe hypoglycaemic episodes.

Theophylline intoxication mimicking diabetic ketoacidosis in a child.

A 5-year-old boy presented with abdominal pain, nausea and vomiting of blood. Twelve hours after admission, "diabetic ketoacidosis" was diagnosed on the basis of elevated glycaemia, glycosuria, ketonuria and a low bicarbonate blood level, which led to treatment with fluids and regular insulin infusion. Over a 36-hour period, insulin was progressively decreased and finally stopped because of the rapid fall and normalization of blood glucose concentration. Drug poisoning was suspected on the basis of persistent tachycardia in the absence of other signs of dehydration. Salicylate intoxication was excluded, and theophylline was finally incriminated. This compound, used by adults in the child's home, had caused accidental theophylline poisoning, mimicking diabetic ketoacidosis. Pre-diabetic immune markers were repeatedly negative, and no diabetes has developed after four years of follow-up. Thus, the transient increase in blood glucose was not related to a pre-diabetic status. A diagnosis of masked theophylline poisoning should be considered in similar situations involving a rapid decrease of insulin requirements.

Xantogranuloma juvenil / Juvenile xanthogranuloma

Presentamos un paciente de 3 y medio años de edad portador de un xantogranuloma juvenil extendido. Las lesiones fueron atipicas desde el punto de vista clinico e histopatologico y uno de los estudios sugirio el diagnostico de histiocitos x (Granuloma de Celulas de Langerhans).El diagnostico final de Xantogranuloma juvenil se establecio sobre la base del curso benigno de la enfermedad,la normalidad de los hallasgos fisicos y de laboratorio y del estudio de la ultraestructura de la biopsia de piel.AU