Failure to bictegravir and development of resistance mutations in an antiretroviral-experienced patient.

We present here one of the first cases of virological failure during treatment with bictegravir/emtricitabine/tenofovir alafenamide (BIC/FTC/TAF). On March 2019, an antiretroviral-experienced HIV-infected patient was admitted to hospital because of cerebral toxoplasmosis. After undergoing treatment with sulfadiazine-pyrimethamine for two weeks, the patient initiated a BIC/FTC/TAF treatment, with 6.01 HIV RNA Log copies/mL, and 37 CD4 cells/µL. After two months under antiretroviral therapy (ART), acute neurologic deterioration with epilepsy, right hemiparesis and dysphagia occurred, leading to nasogastric nutrition and treatment. After several weeks, virological failure was confirmed with 4.01 HIV RNA Log copies/mL and R263K and M184V resistance mutations were detected.

Treatment outcome in dually HIV-1 and HIV-2 coinfected patients living in Spain.

BACKGROUND: Whereas HIV-1 has spread globally, HIV-2 is mainly found in West Africa where dual HIV-1/HIV-2 coinfection is nowadays uncommon. Herein, we report the rate, main characteristics, and treatment outcomes of all dually infected patients living in Spain. METHODS: We identified retrospectively all persons coinfected with HIV-1 recorded at the Spanish HIV-2 registry. Dual infection had been confirmed using PCR in plasma and/or cells, and/or using discriminatory serological tests. RESULTS: From a total of 373 individuals with HIV-2 recorded at the Spanish registry, 34 (9.1%) were coinfected with HIV-1. Compared with HIV-2 monoinfected persons, dually infected patients were more often male (67.6%), presented with lower median CD4 cell counts (204 cells/µl), and had developed more frequently AIDS events (26.5%). Although 61.7% came from West Africa, 6 (17.6%) were native Spaniards. HIV-1 non-B subtypes were recognized in 75% of coinfected patients, being the most prevalent CRF02_AG. At baseline, 45% of dually infected patients had undetectable plasma HIV-2 RNA. After a median follow-up of 32 (13-48) months on antiretroviral therapy, dually infected patients achieved undetectable viremia in 85% for HIV-1, in 80% for HIV-2; and in 70% for both viruses. Median CD4 cell counts reached up to 418 cells/µl. CONCLUSION: Roughly 9% of individuals with HIV-2 infection living in Spain are coinfected with HIV-1. Overall, 70% of dually infected patients achieved viral suppression for both viruses under antiretroviral therapy. Given the relatively large population of West Africans living in Spain and the continuous migration flow from HIV-2 endemic areas, HIV-1/HIV-2 coinfection should always be excluded at first diagnosis in all HIV-seroreactive persons.

Prevalence of resistance associated substitutions and efficacy of baseline resistance-guided chronic hepatitis C treatment in Spain from the GEHEP-004 cohort.

Treatment guidelines differ in their recommendation to determine baseline resistance associated substitutions (RAS) before starting a first-line treatment with direct-acting antivirals (DAAs). Here we analyze the efficacy of DAA treatment with baseline RAS information. We conducted a prospective study involving 23 centers collaborating in the GEHEP-004 DAA resistance cohort. Baseline NS5A and NS3 RASs were studied by Sanger sequencing. After issuing a comprehensive resistance report, the treating physician decided the therapy, duration and ribavirin use. Sustained virological response (SVR12) data are available in 275 patients. Baseline NS5A RAS prevalence was between 4.3% and 26.8% according to genotype, and NS3 RASs prevalence (GT1a) was 6.3%. Overall, SVR12 was 97.8%. Amongst HCV-GT1a patients, 75.0% had >800,000 IU/ml and most of those that started grazoprevir/elbasvir were treated for 12 weeks. In genotype 3, NS5A Y93H was detected in 9 patients. 42.8% of the HCV-GT3 patients that started sofosbuvir/velpatasvir included ribavirin, although only 14.7% carried Y93H. The efficacy of baseline resistance-guided treatment in our cohort has been high across the most prevalent HCV genotypes in Spain. The duration of the grazoprevir/elbasvir treatment adhered mostly to AASLD/IDSA recommendations. In cirrhotic patients infected with GT-3 there has been a high use of ribavirin.

High prevalence and diversity of HIV-1 non-B genetic forms due to immigration in southern Spain: A phylogeographic approach.

Phylogenetic studies are a valuable tool to understand viral transmission patterns and the role of immigration in HIV-1 spread. We analyzed the spatio-temporal relationship of different HIV-1 non-B subtype variants over time using phylogenetic analysis techniques. We collected 693 pol (PR+RT) sequences that were sampled from 2005 to 2012 from naïve patients in different hospitals in southern Spain. We used REGA v3.0 to classify them into subtypes and recombinant forms, which were confirmed by phylogenetic analysis through maximum likelihood (ML) using RAxML. For the main HIV-1 non-B variants, publicly available, genetically similar sequences were sought using HIV-BLAST. The presence of HIV-1 lineages circulating in our study population was established using ML and Bayesian inference (BEAST v1.7.5) and transmission networks were identified. We detected 165 (23.4%) patients infected with HIV-1 non-B variants: 104 (63%) with recombinant viruses in pol: CRF02_AG (71, 43%), CRF14_BG (8, 4.8%), CRF06_cpx (5, 3%) and nine other recombinant forms (11, 6.7%) and unique recombinants (9, 5.5%). The rest (61, 37%) were infected with non-recombinant subtypes: A1 (30, 18.2%), C (7, [4.2%]), D (3, [1.8%]), F1 (9, 5.5%) and G (12, 7.3%). Most patients infected with HIV-1 non-B variants were men (63%, p < 0.001) aged over 35 (73.5%, p < 0.001), heterosexuals (92.2%, p < 0.001), from Africa (59.5%, p < 0.001) and living in the El Ejido area (62.4%, p<0.001). We found lineages of epidemiological relevance (mainly within Subtype A1), imported primarily through female sex workers from East Europe. We detected 11 transmission clusters of HIV-1 non-B Subtypes, which included patients born in Spain in half of them. We present the phylogenetic profiles of the HIV-1 non-B variants detected in southern Spain, and explore their putative geographical origins. Our data reveals a high HIV-1 genetic diversity likely due to the import of viral lineages that circulate in other countries. The highly immigrated El Ejido area acts as a gateway through which different subtypes are introduced into other regions, hence the importance of setting up epidemiological control measures to prevent future outbreaks.

Drug resistance mutations in HIV-2 patients failing raltegravir and influence on dolutegravir response.

Background: A broader extent of amino acid substitutions in the integrase of HIV-2 compared with HIV-1 might enable greater cross-resistance between raltegravir and dolutegravir in HIV-2 infection. Few studies have examined the virological response to dolutegravir in HIV-2 patients that failed raltegravir. Methods: All patients recorded in the HIV-2 Spanish cohort were examined. The integrase coding region was sequenced in viraemic patients. Changes associated with resistance to raltegravir and dolutegravir in HIV-1 were recorded. Results: From 319 HIV-2-infected patients recorded in the HIV-2 Spanish cohort, 53 integrase sequences from 30 individuals were obtained (20 raltegravir naive and 10 raltegravir experienced). Only one secondary mutation (E138A) was found in one of the 20 raltegravir-naive HIV-2 patients. For raltegravir-experienced individuals, the resistance mutation profile in 9 of 10 viraemic patients was as follows: N155H + A153G/S (four); Y143G + A153S (two); Q148R + G140A/S (two); and Y143C + Q91R (one). Of note, all patients with Y143G and N155H developed a rare non-polymorphic mutation at codon 153. Rescue therapy with dolutegravir was given to 5 of these 10 patients. After >6 months on dolutegravir therapy, three patients with baseline N155H experienced viral rebound. In two of them N155H was replaced by Q148K/R and in another by G118R. Conclusions: A wide repertoire of resistance mutations in the integrase gene occur in HIV-2-infected patients failing on raltegravir. Although dolutegravir may allow successful rescue in most HIV-2 raltegravir failures, we report and characterize three cases of dolutegravir resistance in HIV-2 patients, emerging variants Q148K and Q148R and a novel change G118R.

Los estudios de resistencias a antirretrovirales como herramienta para el análisis de los clusters de transmisión del virus de la inmunodeficiencia humana / Study of human immunodeficiency virus transmission chains in Andalusia: Analysis from baseline antiretroviral resistance sequences

INTRODUCCIÓN Y OBJETIVO: Las secuencias de proteasa y transcriptasa reversa del VIH-1 aportan una información muy valiosa para el manejo de la infección por VIH, más allá de la información de resistencias a los antirretrovirales. En nuestro estudio la hemos utilizado para evaluar las cadenas de transmisión, la transmisión de resistencias entre ellos, y para conocer la distribución espacial de los diferentes subtipos utilizando técnicas de georreferenciación. MÉTODOS: Hemos estudiado 693 pacientes diagnosticados de VIH-1 durante el periodo 2005-2012, todos ellos residentes en Andalucía Oriental. La secuencia del gen pol (transcriptasa reversa y proteasa) se generó utilizando Trugene® HIV Genotyping Kit (Siemens, NAD). La historia evolutiva fue inferida a través de MEGA 5.2 mediante el método de Neighbor-Joining. Para la filogeografía y el estudio de resistencias utilizamos ArcGIS y REGA. RESULTADOS: Doscientos noventa y ocho pacientes se asociaron en 77 clusters diferentes. La mayoría de los cluster estaban formados por parejas (n = 49), de hombres que practican sexo con hombres (n = 26), de nacionalidad española (n = 37), con una edad menor a 45 años (73,5%). Las áreas de mayor heterogeneidad de subtipos fueron el área metropolitana de Granada y las zonas de costa de Almería y Granada. Hemos encontrado 5 cluster con más de 10 individuos. En 15 cluster detectamos mutaciones de resistencia. CONCLUSIONES: Presentamos datos que demuestran que el estudio epidemiológico de los diferentes clusters de transmisión de VIH mediante análisis filogenético se presenta como una herramienta potente y de gran utilidad para la vigilancia y control epidemiológico de la propagación del VIH, que puede ayudar a diseñar actuaciones eficaces para prevenir la diseminación del VIH

INTRODUCTION AND OBJECTIVE: Protease and reverse transcriptase HIV-1 sequences provide useful information for patient clinical management, as well as information on resistance to antiretrovirals. The aim of this study is to evaluate transmission events, transmitted drug resistance, and to georeference subtypes among newly diagnosed patients referred to our center. METHODS: A study was conducted on 693 patients diagnosed between 2005 and 2012 in Southern Spain. Protease and reverse transcriptase sequences were obtained for resistance to cART analysis with Trugene® HIV Genotyping Kit (Siemens, NAD). MEGA 5.2, Neighbor-Joining, ArcGIS and REGA were used for subsequent analysis. RESULTS: The results showed 298 patients clustered into 77 different transmission events. Most of the clusters were formed by pairs (n = 49), of men having sex with men (n = 26), Spanish (n = 37), and below 45 years of age (73.5%). Urban areas from Granada, and the coastal areas of Almeria and Granada showed the greatest subtype heterogeneity. Five clusters were formed by more than 10 patients, and 15 clusters had transmitted drug resistance. CONCLUSIONS: The study data demonstrate how the phylogenetic characterization of transmission clusters is a powerful tool to monitor the spread of HIV, and may contribute to design correct preventive measures to minimize it

Lipid profile response to weight loss program in overweight and obese patient is related with gender and age.

INTRODUCTION: The risk of developing cardiovascular disease (CVD) increases with age, however CVD is markedly higher in men than in no-menopausal women. There are few interventions where compare the different effects to lose weight on lipid profile between men and women. OBJECTIVE: The aim of this study was to compare the response on the lipid profile by gender after a weight loss program, and determine whether there are differences by age group. METHODS: One hundred eighty (96 women and 84 men) overweight and obese participants (BMI 25-34.9 kg/m2) aged 18-50 years were randomised into treatment groups. The intervention period was 22 weeks (in all cases 3 times/wk of training for 22 weeks and 2 weeks for pre and post evaluation). All subjects followed a hypocaloric diet (25-30% less energy intake). Energy intake, body composition) and blood lipid profile were recorded at baseline and after of treatment. RESULTS: The response of HDL varied between men and women (p = 0.001). While in women it decreased (HDL: -2.94%, p = 0.02), HDL was elevated in men (HDL: 5% p = 0.02). After intervention men achieved decrease significantly LDL values a 6.65% more than women (p = 0.01). For TG concentrations there were significant differences between men and women in baseline however, only men had a significant chance in post-training measured (p = 0.001). TC showed significant differences between men and women in baseline (p = 0.013). After intervention, men and women showed a significant decreased to TC (p = 0.01). CONCLUSION: Men achieve a positive greater change on lipid profile than women. In addition, the favorable lipid profile response decreases with increasing age.

Introducción: el riesgo de desarrollar una enfermedad cardiovascular (ECV) se incrementa con la edad; sin embargo, el riesgo de ECV en edad fértil es mayor en hombres que en mujeres. Son pocas las intervenciones en las que se comparan las diferencias entre hombres y mujeres que un programa de pérdida de peso tiene sobre el perfil lipídico. Objetivo: comprobar el cambio en el perfil lipídico entre hombres y mujeres tras un programa de pérdida de peso, comparando las diferencias por género y categoría de edad. Métodos: ciento ochenta participantes (96 mujeres y 84 hombres) con sobrepeso y obesidad (IMC 25­34.9 kg/m2) con edades comprendidas entre los 18-50 años fueron repartidos de forma aleatoria en los diferentes grupos de intervención. El período de intervención fue de 22 semanas y 2 semanas para la evaluación pre y post. Todos los sujetos siguieron una dieta equilibrada hipocalórica (25- 30% de restricción calórica) y un programa de ejercicio 3 veces/semana. Antes y después de la intervención todos los grupos fueron evaluados de los cambios en el perfil lipídico, la composición corporal y la ingesta diaria. Resultados: hubo diferencias significativas en el cambio de HDL entre hombres y mujeres (p = 0,001). Mientras que en las mujeres disminuyó (HDL: -2,94%, p = 0,02), en los hombres hubo un aumento de la concentración de HDL (HDL: 5% p = 0,02). Después de la intervención los hombres lograron disminuir significativamente el LDL un 6,65% más que las mujeres (p = 0,01). Para concentraciones de TG hubo diferencias significativas entre hombres y mujeres al inicio de la intervención; sin embargo, solo los hombres tuvieron una mejoría significativa tras la intervención (p = 0,001). El TC mostró diferencias significativas entre hombres y mujeres preintervención (p = 0,013). Tras la intervención, los hombres y las mujeres mostraron una significativa disminución de TC (p = 0,01). Conclusión: los hombres obtuvieron un cambio más favorable en las variables del perfil lipídico con respecto a la mujeres. Además, la respuesta al perfil lipídico favorable disminuye con el aumento de la edad.

Lipid profile response to weight loss program in overweight and obese patient is related with gender and age / Influencia del género y la edad en la respuesta del perfil lipídico tras un programa de pérdida de peso en personas con sobrepeso y obesidad

Introduction: the risk of developing cardiovascular disease (CVD) increases with age, however CVD is markedly higher in men than in no-menopausal women. There are few interventions where compare the different effects to lose weight on lipid profile between men and women. Objective: the aim of this study was to compare the response on the lipid profile by gender after a weight loss program, and determine whether there are differences by age group. Methods: one hundred eighty (96 women and 84 men) overweight and obese participants (BMI 25–34.9 kg/ m2 ) aged 18–50 years were randomised into treatment groups. The intervention period was 22 weeks (in all cases 3 times/wk of training for 22 weeks and 2 weeks for pre and post evaluation). All subjects followed a hypocaloric diet (25-30% less energy intake). Energy intake, body composition) and blood lipid profile were recorded at baseline and after of treatment. Results: the response of HDL varied between men and women (p = 0.001). While in women it decreased (HDL: -2.94%, p = 0.02), HDL was elevated in men (HDL: 5% p = 0.02). After intervention men achieved decrease significantly LDL values a 6.65% more than women (p = 0.01). For TG concentrations there were significant differences between men and women in baseline however, only men had a significant chance in post-training measured (p = 0.001). TC showed significant differences between men and women in baseline (p = 0.013). After intervention, men and women showed a significant decreased to TC (p = 0.01). Conclusion: men achieve a positive greater change on lipid profile than women. In addition, the favorable lipid profile response decreases with increasing age (AU)

Introducción: el riesgo de desarrollar una enfermedad cardiovascular (ECV) se incrementa con la edad; sin embargo, el riesgo de ECV en edad fértil es mayor en hombres que en mujeres. Son pocas las intervenciones en las que se comparan las diferencias entre hombres y mujeres que un programa de pérdida de peso tiene sobre el perfil lipídico. Objetivo: comprobar el cambio en el perfil lipídico entre hombres y mujeres tras un programa de pérdida de peso, comparando las diferencias por género y categoría de edad. Métodos: ciento ochenta participantes (96 mujeres y 84 hombres) con sobrepeso y obesidad (IMC 25–34.9 kg/m2) con edades comprendidas entre los 18-50 años fueron repartidos de forma aleatoria en los diferentes grupos de intervención. El período de intervención fue de 22 semanas y 2 semanas para la evaluación pre y post. Todos los sujetos siguieron una dieta equilibrada hipocalórica (25- 30% de restricción calórica) y un programa de ejercicio 3 veces/semana. Antes y después de la intervención todos los grupos fueron evaluados de los cambios en el perfil lipídico, la composición corporal y la ingesta diaria. Resultados: hubo diferencias significativas en el cambio de HDL entre hombres y mujeres (p = 0,001). Mientras que en las mujeres disminuyó (HDL: -2,94%, p = 0,02), en los hombres hubo un aumento de la concentración de HDL (HDL: 5% p = 0,02). Después de la intervención los hombres lograron disminuir significativamente el LDL un 6,65% más que las mujeres (p = 0,01). Para concentraciones de TG hubo diferencias significativas entre hombres y mujeres al inicio de la intervención; sin embargo, solo los hombres tuvieron una mejoría significativa tras la intervención (p = 0,001). El TC mostró diferencias significativas entre hombres y mujeres preintervención (p = 0,013). Tras la intervención, los hombres y las mujeres mostraron una significativa disminución de TC (p = 0,01). Conclusión: los hombres obtuvieron un cambio más favorable en las variables del perfil lipídico con respecto a la mujeres. Además, la respuesta al perfil lipídico favorable disminuye con el aumento de la edad (AU)

Lipid profile response to weight loss program in overweight and obese patient is related with gender and age / Influencia del género y la edad en la respuesta del perfil lipídico tras un programa de pérdida de peso en personas con sobrepeso y obesidad

Introduction: the risk of developing cardiovascular disease (CVD) increases with age, however CVD is markedly higher in men than in no-menopausal women. There are few interventions where compare the different effects to lose weight on lipid profile between men and women. Objective: the aim of this study was to compare the response on the lipid profile by gender after a weight loss program, and determine whether there are differences by age group. Methods: one hundred eighty (96 women and 84 men) overweight and obese participants (BMI 25–34.9 kg/ m2 ) aged 18–50 years were randomised into treatment groups. The intervention period was 22 weeks (in all cases 3 times/wk of training for 22 weeks and 2 weeks for pre and post evaluation). All subjects followed a hypocaloric diet (25-30% less energy intake). Energy intake, body composition) and blood lipid profile were recorded at baseline and after of treatment. Results: the response of HDL varied between men and women (p = 0.001). While in women it decreased (HDL: -2.94%, p = 0.02), HDL was elevated in men (HDL: 5% p = 0.02). After intervention men achieved decrease significantly LDL values a 6.65% more than women (p = 0.01). For TG concentrations there were significant differences between men and women in baseline however, only men had a significant chance in post-training measured (p = 0.001). TC showed significant differences between men and women in baseline (p = 0.013). After intervention, men and women showed a significant decreased to TC (p = 0.01). Conclusion: men achieve a positive greater change on lipid profile than women. In addition, the favorable lipid profile response decreases with increasing age (AU)

Introducción: el riesgo de desarrollar una enfermedad cardiovascular (ECV) se incrementa con la edad; sin embargo, el riesgo de ECV en edad fértil es mayor en hombres que en mujeres. Son pocas las intervenciones en las que se comparan las diferencias entre hombres y mujeres que un programa de pérdida de peso tiene sobre el perfil lipídico. Objetivo: comprobar el cambio en el perfil lipídico entre hombres y mujeres tras un programa de pérdida de peso, comparando las diferencias por género y categoría de edad. Métodos: ciento ochenta participantes (96 mujeres y 84 hombres) con sobrepeso y obesidad (IMC 25–34.9 kg/m2) con edades comprendidas entre los 18-50 años fueron repartidos de forma aleatoria en los diferentes grupos de intervención. El período de intervención fue de 22 semanas y 2 semanas para la evaluación pre y post. Todos los sujetos siguieron una dieta equilibrada hipocalórica (25- 30% de restricción calórica) y un programa de ejercicio 3 veces/semana. Antes y despueìs de la intervencioìn todos los grupos fueron evaluados de los cambios en el perfil lipiìdico, la composicioìn corporal y la ingesta diaria. Resultados: hubo diferencias significativas en el cambio de HDL entre hombres y mujeres (p = 0,001). Mientras que en las mujeres disminuyó (HDL: -2,94%, p = 0,02), en los hombres hubo un aumento de la concentración de HDL (HDL: 5% p = 0,02). Después de la intervención los hombres lograron disminuir significativamente el LDL un 6,65% más que las mujeres (p = 0,01). Para concentraciones de TG hubo diferencias significativas entre hombres y mujeres al inicio de la intervención; sin embargo, solo los hombres tuvieron una mejoría significativa tras la intervención (p = 0,001). El TC mostró diferencias significativas entre hombres y mujeres preintervención (p = 0,013). Tras la intervención, los hombres y las mujeres mostraron una significativa disminución de TC (p = 0,01). Conclusión: los hombres obtuvieron un cambio más favorable en las variables del perfil lipídico con respecto a la mujeres. Además, la respuesta al perfil lipídico favorable disminuye con el aumento de la edad (AU)

Dolutegravir for the treatment of HIV-2 infection.

BACKGROUND: Therapeutic options are limited for HIV-2 infected persons, largely in part due to the lack of susceptibility to HIV-1 non-nucleoside reverse transcriptase inhibitors and poor susceptibility to some HIV-1 protease inhibitors. This is particularly worrisome for HIV-2 patients with prior antiretroviral failure. OBJECTIVES: Report the virological response to dolutegravir in HIV-2-infected individuals. STUDY DESIGN: Retrospective observational assessment of all HIV-2 individuals treated with dolutegravir in Spain. RESULTS: From 297 HIV-2-infected individuals recorded at the Spanish national registry, 26% received antiretroviral therapy. Six out of 8 failing on raltegravir selected for integrase resistance mutations N155H (4), Y143G (1) and Q148R (1). Two patients bearing N155H subsequently received dolutegravir. Both experienced initially more than 1.5 log drop in plasma HIV-2 RNA and significant CD4 gains. Whereas one kept on undetectable viremia 6 months later, the other experienced viral rebound. CONCLUSION: Dolutegravir may be a good therapeutic option for patients with HIV-2 infection, including those that previously failed other integrase inhibitors.

[Study of human immunodeficiency virus transmission chains in Andalusia: analysis from baseline antiretroviral resistance sequences]. / Los estudios de resistencias a antirretrovirales como herramienta para el análisis de los clusters de transmisión del virus de la inmunodeficiencia humana.

INTRODUCTION AND OBJECTIVE: Protease and reverse transcriptase HIV-1 sequences provide useful information for patient clinical management, as well as information on resistance to antiretrovirals. The aim of this study is to evaluate transmission events, transmitted drug resistance, and to georeference subtypes among newly diagnosed patients referred to our center. METHODS: A study was conducted on 693 patients diagnosed between 2005 and 2012 in Southern Spain. Protease and reverse transcriptase sequences were obtained for resistance to cART analysis with Trugene(®) HIV Genotyping Kit (Siemens, NAD). MEGA 5.2, Neighbor-Joining, ArcGIS and REGA were used for subsequent analysis. RESULTS: The results showed 298 patients clustered into 77 different transmission events. Most of the clusters were formed by pairs (n=49), of men having sex with men (n=26), Spanish (n=37), and below 45 years of age (73.5%). Urban areas from Granada, and the coastal areas of Almeria and Granada showed the greatest subtype heterogeneity. Five clusters were formed by more than 10 patients, and 15 clusters had transmitted drug resistance. CONCLUSIONS: The study data demonstrate how the phylogenetic characterization of transmission clusters is a powerful tool to monitor the spread of HIV, and may contribute to design correct preventive measures to minimize it.

Systematic production of inactivating and non-inactivating suppressor mutations at the relA locus that compensate the detrimental effects of complete spot loss and affect glycogen content in Escherichia coli.

In Escherichia coli, ppGpp is a major determinant of growth and glycogen accumulation. Levels of this signaling nucleotide are controlled by the balanced activities of the ppGpp RelA synthetase and the dual-function hydrolase/synthetase SpoT. Here we report the construction of spoT null (ΔspoT) mutants obtained by transducing a ΔspoT allele from ΔrelAΔspoT double mutants into relA+ cells. Iodine staining of randomly selected transductants cultured on a rich complex medium revealed differences in glycogen content among them. Sequence and biochemical analyses of 8 ΔspoT clones displaying glycogen-deficient phenotypes revealed different inactivating mutations in relA and no detectable ppGpp when cells were cultured on a rich complex medium. Remarkably, although the co-existence of ΔspoT with relA proficient alleles has generally been considered synthetically lethal, we found that 11 ΔspoT clones displaying high glycogen phenotypes possessed relA mutant alleles with non-inactivating mutations that encoded stable RelA proteins and ppGpp contents reaching 45-85% of those of wild type cells. None of the ΔspoT clones, however, could grow on M9-glucose minimal medium. Both Sanger sequencing of specific genes and high-throughput genome sequencing of the ΔspoT clones revealed that suppressor mutations were restricted to the relA locus. The overall results (a) defined in around 4 nmoles ppGpp/g dry weight the threshold cellular levels that suffice to trigger net glycogen accumulation, (b) showed that mutations in relA, but not necessarily inactivating mutations, can be selected to compensate total SpoT function(s) loss, and (c) provided useful tools for studies of the in vivo regulation of E. coli RelA ppGpp synthetase.

EasyLCMS: an asynchronous web application for the automated quantification of LC-MS data.

BACKGROUND: Downstream applications in metabolomics, as well as mathematical modelling, require data in a quantitative format, which may also necessitate the automated and simultaneous quantification of numerous metabolites. Although numerous applications have been previously developed for metabolomics data handling, automated calibration and calculation of the concentrations in terms of µmol have not been carried out. Moreover, most of the metabolomics applications are designed for GC-MS, and would not be suitable for LC-MS, since in LC, the deviation in the retention time is not linear, which is not taken into account in these applications. Moreover, only a few are web-based applications, which could improve stand-alone software in terms of compatibility, sharing capabilities and hardware requirements, even though a strong bandwidth is required. Furthermore, none of these incorporate asynchronous communication to allow real-time interaction with pre-processed results. FINDINGS: Here, we present EasyLCMS (http://www.easylcms.es/), a new application for automated quantification which was validated using more than 1000 concentration comparisons in real samples with manual operation. The results showed that only 1% of the quantifications presented a relative error higher than 15%. Using clustering analysis, the metabolites with the highest relative error distributions were identified and studied to solve recurrent mistakes. CONCLUSIONS: EasyLCMS is a new web application designed to quantify numerous metabolites, simultaneously integrating LC distortions and asynchronous web technology to present a visual interface with dynamic interaction which allows checking and correction of LC-MS raw data pre-processing results. Moreover, quantified data obtained with EasyLCMS are fully compatible with numerous downstream applications, as well as for mathematical modelling in the systems biology field.

[Hepatitis B virus (HBV) infection in Sub-Saharan immigrants in Almería (Spain)]. / Infección por virus de hepatitis B (VHB) en inmigrantes subsaharianos en Almería.

INTRODUCTION: There is scant information available in Spain regarding virological markers and clinical status in Sub-Saharan patients infected with HVB. METHODS: A cross-sectional and retrospective study of virological markers and clinical status of HBV infection in 510 adult patients from Sub-Saharan Africa, not co-infected with HIV, most of them from West Africa countries. RESULTS: A total of 90.8% of patients had markers of HBV infection and 137 (26.9%) were HBsAg positive. Among patients with HBsAg positive, 55.9% were chronic inactive carriers. The predominant genotype was E. CONCLUSIONS: The study shows a high prevalence of both markers of HBV infection and of chronic hepatitis B in immigrants from Sub-Saharan Africa.

Infección por virus de hepatitis B (VHB) en inmigrantes subsaharianos en Almería / Hepatitis B virus (HBV) infection in Sub-Saharan immigrants in Almería (Spain)

Introducción Existen escasos estudios en España sobre la situación virológica y clínica de los pacientes subsaharianos infectados por VHB. Métodos Estudio transversal y retrospectivo de marcadores virológicos y clínico de infección por VHB en 510 pacientes subsaharianos adultos, no infectados por VIH, procedentes en su mayoría del Oeste de África. Resultados El 90,8% presentaba marcadores de infección por VHB. Ciento treinta y siete pacientes (26,9%) presentaban AgHBs+, siendo el 55,9% portadores crónicos inactivos. El genotipo E fue el predominante. Conclusiones El estudio demuestra una elevada prevalencia de marcadores serológicos y sobre todo de infección crónica por VHB en la población inmigrante subsahariana (AU)

Introduction There is scant information available in Spain regarding virological markers and clinical status in Sub-Saharan patients infected with HVB. Methods A cross-sectional and retrospective study of virological markers and clinical status of HBV infection in 510 adult patients from Sub-Saharan Africa, not co-infected with HIV, most of them from West Africa countries .Results A total of 90.8% of patients had markers of HBV infection and 137 (26.9%) were HBsAg positive. Among patients with HBsAg positive, 55.9% were chronic inactive carriers. The predominant genotype was E. Conclusions The study shows a high prevalence of both markers of HBV infection and of chronic hepatitis B in immigrants from Sub-Saharan Africa (AU)