Deep analysis of HIV-1 natural variability across HIV-1 variants at residues associated with integrase inhibitor (INI) resistance in INI-naive individuals.

OBJECTIVES: We evaluated variant-associated variability at positions related to resistance to the integrase (IN) inhibitors (INIs) raltegravir, elvitegravir and dolutegravir using HIV-1 IN sequences from naive individuals retrieved from GenBank. METHODS: We evaluated the frequency of major, secondary and rare amino acid changes associated with INI resistance (INI-R) in 6706 sequences from 3791 INI-naive individuals carrying a large panel of different HIV-1 variants retrieved from GenBank, including four groups: M (6663), O (24), N (15) and P (4). HIV-1 group M sequences included 4599 sequences from the nine group M subtypes and 2064 recombinants ascribed to 54 circulating recombinant forms (CRFs). RESULTS: Primary INI-R mutations were rare in INI-naive participants and only present at a low rate in subtypes B, C and D and recombinants CRF01_AE and CRF14_BG, ranging from one to five per variant. Three secondary INI-R changes appeared with variable frequency in INI-naive individuals carrying specific HIV-1 variants: L74M in CRF43_02G (33.3%); T97A in group P (50%), J (33.3%), CRF18_cpx (20%) and F2 (11.5%); and G163RK in CRF44_BF (100%), CRF46_BF (66.7%), CRF17_BF (28.6%), F1 (21.7%), CRF12_BF (16.7%) and CRF29_BF (12.5%). Rare mutations were absent. CONCLUSIONS: Natural variability in INI-R positions across HIV-1 variants should be studied as they may facilitate or delay the emergence of INI-R viruses.

Time-dependent changes in the expression of lymphocyte and monocyte cell adhesion molecules after meals of different composition.

The objective of the present study was to compare the acute effect of meals of different composition on the expression of adhesion molecules that play a key role in leucocyte trafficking. A total of twenty apparently healthy subjects randomly consumed three isoenergetic meals 1 week apart: enriched in carbohydrates (CHO), enriched in monounsaturated fat and enriched in saturated fat. Blood samples were obtained before the meals and at 2, 4, 6, 8 and 10 h after meal ingestion. Samples were analysed for LDL resistance to Cu-mediated oxidation and for the surface expression on peripheral blood mononuclear cells (PBMC) of CD62L, CD162, CD11a, CD11b, CD49d and CD54 by flow cytometry. The present results showed that there were no changes in LDL susceptibility to oxidation within and among the meals. After the CHO-enriched meal, there was a time-dependent increased expression of CD162, CD49d, CD11a and CD54 on PBMC that returned to basal values after 8-10 h. These changes were significantly greater than the ones observed after the consumption of the monounsaturated fat- and the saturated fat-enriched meals and were more evident in lymphocytes than in monocytes. In conclusion, acute ingestion of a CHO-enriched meal induces higher increases of lymphocyte activation markers than fat-enriched meals. These results suggest that long-term consumption of CHO-enriched diets may be associated with a sustained pro-inflammatory state.

[Clinical factors associated with inappropriate prescription of statins]. / Prescripción inadecuada de estatinas y factores clínicos asociados.

BACKGROUND AND OBJECTIVE: The aim of this study was to determine the percentage of subjects receiving statin treatment that was inappropriate and to evaluate the associated clinical factors. PATIENTS AND METHOD: A cross-sectional multi-centred study conducted across the Autonomous Regions of Spain. The appropriateness of prescription was based on the guidelines of the National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP III) and the European Guidelines on Cardiovascular Disease Prevention. RESULTS: Of the 1,817 patients studied, 52.9% were male, the overall mean age was 59.4 years, 36% had cardiovascular disease and 32.7% had diabetes. In 90.8% of the population the prescription was correct according to at least one of the two guides being used, with a low concordance between them (kappa = 0.279). Almost 100% of subjects with cardiovascular disease or diabetes had an appropriate prescription. In primary prevention, 82.7% had an appropriate prescription according to at least one of the two guides used, but this percentage decreased to 20.7% when based on the European Guidelines. In multivariate analysis, there was agreement with both guides. The inappropriate prescription was directly, and significantly, associated with primary prevention and female gender, while being inversely related to age, smoking habit and LDL-cholesterol concentration. CONCLUSIONS: Most of the patients receiving statin treatment have an appropriate prescription, but the percentage of inappropriate prescription increases significantly in patients in primary prevention following European Guidelines.

New findings in cleavage sites variability across groups, subtypes and recombinants of human immunodeficiency virus type 1.

BACKGROUND: Polymorphisms at cleavage sites (CS) can influence Gag and Pol proteins processing by the viral protease (PR), restore viral fitness and influence the virological outcome of specific antiretroviral drugs. However, data of HIV-1 variant-associated CS variability is scarce. METHODS: In this descriptive research, we examine the effect of HIV-1 variants on CS conservation using all 9,028 gag and 3,906 pol HIV-1 sequences deposited in GenBank, focusing on the 110 residues (10 per site) involved at 11 CS: P17/P24, P24/P2, P2/P7, P7/P1, P1/P6 (gag) , NC/TFP, TFP/P6 (pol), P6 (pol) /PR, PR/RT(p51), RT(p51)/RT(p66) and RT(p66)/IN. CS consensus amino acid sequences across HIV-1 groups (M, O, N, P), group M 9 subtypes and 51 circulating recombinant forms (CRF) were inferred from our alignments and compared to the HIV-1 consensus-of-consensuses sequence provided by GenBank. RESULTS: In all HIV-1 variants, the most conserved CS were PR/RT(p51), RT(p51)/RT(p66), P24/P2 and RT(p66)/IN and the least P2/P7 and P6 (pol) /PR. Conservation was significantly lower in subtypes vs. recombinants in P2/P7 and TFP/P6 (pol) and higher in P17/P24. We found a significantly higher conservation rate among Group M vs. non-M Groups HIV-1. The late processing sites at Gag (P7/P1) and GagPol precursors (PR/RT(p51)) presented a significantly higher conservation vs. the first CS (P2/P7) in the 4 HIV-1 groups. Here we show 52 highly conserved residues across HIV-1 variants in 11 CS and the amino acid consensus sequence in each HIV-1 group and HIV-1 group M variant for each 11 CS. CONCLUSIONS: This is the first study to describe the CS conservation level across all HIV-1 variants and 11 sites in one of the largest available sequence HIV-1 dataset. These results could help other researchers for the future design of both novel antiretroviral agents acting as maturation inhibitors as well as for vaccine targeting CS.

Liver upregulation of genes involved in cortisol production and action is associated with metabolic syndrome in morbidly obese patients.

BACKGROUND: Hepatic 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) activity, which converts cortisone (inactive) to cortisol, is downregulated in obesity. However, this compensation fails in obese with metabolic abnormalities, such as diabetes. To further characterize the tissue-specific cortisol regeneration in obesity, we have investigated the mRNA expression of genes related to local cortisol production, i.e., 11ß-HSD1, hexose-6-phosphate dehydrogenase (H6PDH) and cortisol action, glucocorticoid receptor (GR) and a cortisol target gene, phosphoenolpyruvate carboxykinase (PEPCK) in the liver, and visceral (VAT) and subcutaneous (SAT) adipose tissues from morbidly obese patients with and without metabolic syndrome (MS). METHODS: Fifty morbidly obese patients undergoing bariatric surgery, 14 men (mean age, 41.3 ± 3.5 years; BMI, 48.0 ± 3.6 kg/m(2)) and 36 women (mean age, 44.6 ± 1.9 years; BMI, 44.9 ± 1.2 kg/m(2)), were classified as having MS (MS+, n = 20) or not (MS-, n = 30). Tissue mRNA levels were measured by real-time polymerase chain reaction. RESULTS: Hepatic mRNA levels of these genes were higher in obese patients with MS (11ß-HSD1, P = 0.002; H6PDH, P = 0.043; GR, P = 0.033; PEPCK, P = 0.032) and positively correlated with the number of clinical characteristics that define the MS. The expression of the four genes positively correlated among them. In contrast to the liver, these genes were not differently expressed in VAT or SAT, when MS+ and MS- obese patients were compared. CONCLUSIONS: Coordinated liver-specific upregulation of genes involved in local cortisol regeneration and action support the concept that local hepatic hypercortisolism contributes to development of MS in morbidly obese patients.

Modificaciones en la activación y la expresión de moléculas de adhesión y receptores celulares monocitarios en pacientes hipercolesterolémicos tratados con pravastatina / Changes in the activation and expression of monocyte adhesion molecules and receptors in hypercholesterolemic patients treated with pravastatin

Introducción. Numerosos estudios han demostrado que las estatinas reducen la mortalidad cardiovascular. Si bien la mayor parte del efecto es probablemente debido a su acción hipolipemiante, se han descrito otras acciones de estos fármacos, independientes de la reducción del colesterol, que podrían contribuir a este efecto beneficioso. El objetivo del presente estudio fue evaluar el efecto del tratamiento con estatinas sobre la expresión de moléculas de adhesión, citocinas y receptores celulares de monocitos de sangre periférica, células estrechamente ligadas al desarrollo de la arteriosclerosis. Pacientes y métodos. Se seleccionó a 22 pacientes con hipercolesterolemia moderada (59% mujeres, edad media 51 años). Tras seguir durante 6 semanas una dieta baja en grasas, se aleatorizaron a recibir 40 mg/día de pravastatina o placebo durante 8 semanas, tras lo cual recibieron durante otras 8 semanas el tratamiento contrario. Resultados. El tratamiento con pravastatina redujo de manera significativa, respecto al período placebo, la concentración de colesterol total (22%; p < 0,01) y el colesterol unido a lipoproteínas de baja densidad (30%; p < 0,01). La expresión intracitoplasmática monocitaria del factor de necrosis tumoral alfa (TNFα) se redujo en un 17,3% (p = 0,03). No se observaron modificaciones en la expresión de CD62L, CD162, CD11a, CD11b, CD49d, CD54, MCP-1 y CCR2. Conclusión. El tratamiento con pravastatina durante 8 semanas en pacientes con hipercolesterolemia moderada induce un descenso en la intensidad media de fluorescencia con la que los monocitos de sangre periférica expresan TNFα, sin modificar la expresión de otras moléculas de adhesión (AU)

Introduction. Numerous studies have shown that statins reduce cardiovascular mortality. While most of the effect is probably due to their lipid-lowering action, other actions of these drugs, independent of cholesterol reduction, have been described that could influence these beneficial action. The aim of this study was to evaluate the effect of statins on the expression of adhesion molecules, cytokines and receptors of peripheral blood monocytes, cells closely linked to development of atherosclerosis. Patients and methods. We selected 22 patients with moderate hypercholesterolemia (59% female, mean age 51 years). After 6 weeks of a low fat they were randomized to receive 40 mg of pravastatin or placebo for 8 weeks. After that period they received for another 8 weeks the other treatment. Results. Treatment with pravastatin significantly reduced the concentration of total cholesterol (22%; p < 0.01) and LDL-cholesterol (30%; p < 0.01). Monocytic intracytoplasmic expression of TNFα was reduced by 17.3% (p = 0,03). Other cellular markers, CD62L, CD162, CD11a, CD11b, CD49d, CD54, MCP-1 and CCR2, did not change their expression Conclusion. Treatment with pravastatin for 8 weeks in patients with moderate hypercholesterolemia induces a decline in the average intensity of fluorescence with which peripheral blood monocytes express TNFα without changing the expression of other adhesion molecules (AU)

Prescripción inadecuada de estatinas y factores clínicos asociados / Clinical factors associated with inappropriate prescription of statins

Fundamento y objetivo: El objetivo del estudio fue evaluar el porcentaje de sujetos en tratamiento con estatinas con una inadecuada prescripción, así como estudiar los factores asociados a ésta. Pacientes y método: Estudio transversal y multicéntrico en el que participaron 1.817 sujetos en tratamiento con estatinas procedentes de todas las comunidades autónomas. Se evaluó la indicación del tratamiento hipolipemiante según el National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP III) y las Guías Europeas para la Prevención de la Enfermedad Cardiovascular. Resultados: El 52,9% eran varones y la edad media fue de 59,4 años. Un 36,0% presentaba enfermedad cardiovascular y un 32,7% eran diabéticos. En un 90,8% del total de la población estaba correctamente indicado el tratamiento según al menos una de las 2 guías, con una baja concordancia entre ellas (* = 0,279). Aproximadamente en el 100% de los sujetos con enfermedad cardiovascular o diabetes la prescripción era adecuada. En prevención primaria, el tratamiento era adecuado en el 82,7% según al menos una de las 2 guías, si bien este porcentaje descendía hasta el 20,7% al aplicar las Guías Europeas. En el análisis multivariante ambas guías coincidían en que la prescripción inadecuada se asociaba directa y significativamente con estar en prevención primaria y el sexo femenino, e inversamente con la edad, el consumo de tabaco y la concentración de colesterol unido a lipoproteínas de baja densidad (cLDL). Conclusiones: En la mayoría de los pacientes tratados con estatinas la prescripción es adecuada, si bien el porcentaje de tratamientos inadecuados se incrementa sustancialmente en los pacientes en prevención primaria, especialmente con las Guías Europeas

Background and objective: The aim of this study was to determine the percentage of subjects receiving statin treatment that was inappropriate and to evaluate the associated clinical factors. Patients and method: A cross-sectional multi-centred study conducted across the Autonomous Regions of Spain. The appropriateness of prescription was based on the guidelines of the National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATP III) and the European Guidelines on Cardiovascular Disease Prevention. Results: Of the 1,817 patients studied, 52.9% were male, the overall mean age was 59.4 years, 36% had cardiovascular disease and 32.7% had diabetes. In 90.8% of the population the prescription was correct according to at least one of the two guides being used, with a low concordance between them (* = 0.279). Almost 100% of subjects with cardiovascular disease or diabetes had an appropriate prescription. In primary prevention, 82.7% had an appropriate prescription according to at least one of the two guides used, but this percentage decreased to 20.7% when based on the European Guidelines. In multivariate analysis, there was agreement with both guides. The inappropriate prescription was directly, and significantly, associated with primary prevention and female gender, while being inversely related to age, smoking habit and LDL-cholesterol concentration. Conclusions: Most of the patients receiving statin treatment have an appropriate prescription, but the percentage of inappropriate prescription increases significantly in patients in primary prevention following European Guidelines