Soluble Free, Esterified and Insoluble-Bound Phenolic Antioxidants from Chickpeas Prevent Cytotoxicity in Human Hepatoma HuH-7 Cells Induced by Peroxyl Radicals.

Chickpeas are rich sources of bioactive compounds such as phenolic acids, flavonoids, and isoflavonoids. However, the contribution of insoluble-bound phenolics to their antioxidant properties remains unclear. Four varieties of chickpeas were evaluated for the presence of soluble (free and esterified) and insoluble-bound phenolics as well as their antiradical activity, reducing power and inhibition of peroxyl-induced cytotoxicity in human HuH-7 cells. In general, the insoluble-bound fraction showed a higher total phenolic content. Phenolic acids, flavonoids, and isoflavonoids were identified and quantified by UPLC-MS/MS. Taxifolin was identified for the first time in chickpeas. However, m-hydroxybenzoic acid, taxifolin, and biochanin A were the main phenolics found. Biochanin A was mostly found in the free fraction, while m-hydroxybenzoic acid was present mainly in the insoluble-bound form. The insoluble-bound fraction made a significant contribution to the reducing power and antiradical activity towards peroxyl radical. Furthermore, all extracts decreased the oxidative damage of human HuH-7 cells induced by peroxyl radicals, thus indicating their hepatoprotective potential. This study demonstrates that the antioxidant properties and bioactive potential of insoluble-bound phenolics of chickpeas should not be neglected.

Bottom-up analysis of emergent properties of N-acetylcysteine as an adjuvant therapy for COVID-19.

N-acetylcysteine (NAC) is an abundantly available antioxidant with a wide range of antidotal properties currently best studied for its use in treating acetaminophen overdose. It has a robustly established safety profile with easily tolerated side effects and presents the Food and Drug Administration's approval for use in treating acetaminophen overdose patients. It has been proven efficacious in off-label uses, such as in respiratory diseases, heart disease, cancer, human immunodeficiency virus infection, and seasonal influenza. Clinical trials have recently shown that NAC's capacity to replenish glutathione stores may significantly improve coronavirus disease 2019 (COVID-19) outcomes, especially in high risk individuals. Interestingly, individuals with glucose 6-phosphate dehydrogenase deficiency have been shown to experience even greater benefit. The same study has concluded that NAC's ability to mitigate the impact of the cytokine storm and prevent elevation of liver enzymes, C-reactive protein, and ferritin is associated with higher success rates weaning from the ventilator and return to normal function in COVID-19 patients. Considering the background knowledge of biochemistry, current uses of NAC in clinical practice, and newly acquired evidence on its potential efficacy against COVID-19, it is worthwhile to investigate further whether this agent can be used as a treatment or adjuvant for COVID-19.

Frecuencia de hemoglobinopatías en mujeres embarazadas del programa de anemias por hematíes falciformes en Cuba / Frequency of hemoglobinopathies among pregnant women from the sickle cell anemia program in Cuba

Introducción: Las hemoglobinopatías se consideran errores monogénicos hereditarios y están caracterizados por defectos en la molécula de hemoglobina. En Cuba, la detección prenatal de hemoglobinopatías se realiza a través de la electroforesis de hemoglobina para identificar parejas de alto riesgo. El programa brinda: asesoramiento genético, diagnóstico prenatal molecular e interrupciones selectivas de fetos afectados, a solicitud de las parejas. Objetivo: Determinar la frecuencia de hemoglobinopatías en mujeres embarazadas residentes en Cuba. Métodos: Se realizó un estudio descriptivo, retrospectivo y de corte transversal para determinar la frecuencia de hemoglobinopatías en 1 342 917 mujeres embarazadas captadas en el periodo 2009-2019. El método diagnóstico de la pesquisa fue la electroforesis de hemoglobina en geles de agarosa a pH alcalino. La confirmación se realizó por electroforesis de hemoglobina en gel de agarosa a pH ácido; ambos métodos mediante la tecnología HYDRASYS. Resultados: La frecuencia global de embarazadas con hemoglobinopatías fue de 3,5 por ciento. Se detectó hemoglobinopatías en 47 465 mujeres; 38 698 con variante S heterocigoto, 8 706 variantes de hemoglobina C y 158 de otras variantes. Se detectaron 44 283 esposos con hemoglobinopatías, 3 099 parejas de alto riesgo y se realizaron 2 689 diagnósticos prenatales moleculares. Se confirmaron 522 fetos afectados y 382 parejas solicitaron la interrupción del embarazo. El subprograma alcanzó 99,24 por ciento de cobertura en el país. Conclusión: La alta frecuencia de hemoglobinopatías en Cuba justifica la importancia de continuar el subprograma de detección de portadores para prevenir la aparición de las formas graves de la enfermedad(AU)

Introduction: Hemoglobinopathies are hereditary monogenic errors characterized by defects in the hemoglobin molecule. In Cuba, prenatal detection of hemoglobinopathies is performed by hemoglobin electrophoresis to identify high-risk couples. The program offers genetic counseling, prenatal molecular diagnosis and selective pregnancy termination in case of affected fetuses at the request of couples. Objective: Determine the frequency of hemoglobinopathies among pregnant women living in Cuba. Methods: A descriptive cross-sectional retrospective study was conducted to determine the frequency of hemoglobinopathies in 1 342 917 pregnant women recruited in the period 2009-2019. Screening was based on the diagnostic method of hemoglobin electrophoresis in alkaline pH agarose gels. Confirmation was performed with hemoglobin electrophoresis in acid pH agarose gel. Both methods used HYDRASYS technology. Results: Overall frequency of pregnant women with hemoglobinopathies was 3.5 percent. Hemoglobinopathies were detected in 47 465 women: 38 698 with variant S heterozygote, 8 706 with variants of hemoglobin C y 158 with other variants. 44 283 husbands with hemoglobinopathies and 3 099 high-risk couples were detected, and 2 689 prenatal molecular diagnostic tests were conducted. A total 522 affected fetuses were confirmed, and 382 couples requested pregnancy termination. The subprogram achieved 99.24 percent coverage in the country. Conclusion: The high frequency of hemoglobinopathies in Cuba justifies the importance of continuing the carrier detection subprogram to prevent the emergence of severe forms of the disease(AU)

Omega 3 Fatty Acids and COVID-19: A Comprehensive Review.

The rapid international spread of severe acute respiratory syndrome coronavirus 2 responsible for coronavirus disease 2019 (COVID-19) has posed a global health emergency in 2020. It has affected over 52 million people and led to over 1.29 million deaths worldwide, as of November 13th, 2020. Patients diagnosed with COVID-19 present with symptoms ranging from none to severe and include fever, shortness of breath, dry cough, anosmia, and gastrointestinal abnormalities. Severe complications are largely due to overdrive of the host immune system leading to "cytokine storm". This results in disseminated intravascular coagulation, acute respiratory distress syndrome, multiple organ dysfunction syndrome, and death. Due to its highly infectious nature and concerning mortality rate, every effort has been focused on prevention and creating new medications or repurposing old treatment options to ameliorate the suffering of COVID-19 patients including the immune dysregulation. Omega-3 fatty acids are known to be incorporated throughout the body into the bi-phospholipid layer of the cell membrane leading to the production of less pro-inflammatory mediators compared to other fatty acids that are more prevalent in the Western diet. In this article, the benefits of omega-3 fatty acids, especially eicosapentaenoic acid and docosahexaenoic acid, including their anti-inflammatory, immunomodulating, and possible antiviral effects have been discussed.

Diagnóstico de aciduria metilmalónica en el periodo 2013-2018 / Diagnosis of Methyl Malonicaciduria from 2013 to 2018

RESUMEN Fundamento: la aciduria metilmalónica es una de las acidurias orgánicas más frecuentes y agrupa un conjunto de defectos genéticos caracterizados por la excreción de niveles elevados de ácido metilmalónico en la orina. La excreción de este metabolito puede ir acompañada o no de niveles elevados de homocisteína en dependencia de la ruta metabólica afectada. Objetivo describir la implementación de una metodología de laboratorio que combina el ácido metilmalónico y la homocisteína en el diagnóstico diferencial y seguimiento de la aciduria metilmalónica en el periodo de 2013 a 2018. Métodos: a los pacientes con incremento de ácido metilmalónico en el perfil de ácidos orgánicos, se les cuantificó homocisteína en plasma y orina. La identificación del ácido metilmalónico se realizó por cromatografía gaseosa/ espectrometría de masas, mientras que la cuantificación de homocisteína por cromatografía líquida de alta resolución. Resultados: los métodos cromatográficos permitieron la identificación y cuantificación del ácido metilmalónico y la homocisteína, respectivamente. La homocisteína se cuantificó en siete pacientes con niveles incrementados de aciduria metilmalónica. Los niveles de homocisteína en cuatro de ellos fueron superiores a los valores normales, sugiriendo una aciduria combinada con homocistinuria. Tres de los pacientes con aciduria metilmalónica combinada bajo tratamiento mostraron una disminución en los niveles de ambos metabolitos, correspondiendo con una satisfactoria evolución. Conclusiones: la metodología implementada con los análisis de la determinación simultánea de ambos marcadores permitió el diagnóstico diferencial y seguimiento bioquímico de la aciduria metilmalónica.

ABSTRACT Foundation: methylmalonic aciduria is one of the most frequent organic acidurias and groups together a set of genetic defects, characterized by the excretion of elevated levels of urinemethyl malonic acid. The excretion of this metabolite may or may not be accompanied by elevated homocysteine ​​levels depending on the affected metabolic pathway. Objective: to describe the implementation of a laboratory methodology that combines methylmalonic acid and homocysteine ​​in the differential diagnosis and monitoring of methylmalonic aciduria in the period from 2013 to 2018. Methods: for patients with an increase in methylmalonic acid in the organic acid profile, homocysteine ​​was quantified in plasma and urine. The identification of methylmalonic acid was performed by gas chromatography / mass spectrometry, while the homocysteine ​​quantification by high performance liquid chromatography. Results: chromatographic methods allowed the identification and quantification of methylmalonic acid and homocysteine, respectively. Homocysteine ​​was quantified in seven patients with increased levels of methylmalonic aciduria. Homocysteine ​​levels in four of them were higher than normal values, suggesting aciduria combined with homocystinuria. Three of the patients with combined methylmalonic aciduria under treatment showed a decrease in the levels of both metabolites, corresponding to a satisfactory evolution. Conclusions: simultaneous determination of both markers allowed differential diagnosis and biochemical monitoring of this disease.

Material de apoyo para la docencia "Cuidados postoperatorios cardiovasculares en adultos y niños" / Teaching aid for teaching "Cardiovascular postoperative cares in adults and children"

RESUMEN Fundamento: el desarrollo vertiginoso de la ciencia y la técnica incorpora constantemente nuevos adelantos al quehacer diario del ejercicio médico, lo cual reclama una continua capacitación. Esta necesidad se acentúa cuando se trata de la atención a pacientes graves en el contexto del postoperatorio de cirugía cardiovascular, donde la mayoría son atendidos en unidades de terapia intensiva. Objetivos: diseñar un material docente de apoyo bibliográfico para los temas incluidos en el Módulo 25: Cuidados postoperatorios cardiovasculares en el adulto y el niño, del programa de estudio de tercer año de la residencia de Cardiología. Métodos: se realizó una investigación de desarrollo en el Cardiocentro "Ernesto Che Guevara", de Villa Clara, durante diciembre 2017-mayo 2018. Se emplearon métodos teóricos: análisis-síntesis, inducción-deducción y sistémico-estructural; empíricos: análisis documental y cuestionario a docentes médicos especialistas; y matemáticos estadísticos para los valores absolutos y relativos. Resultados: la literatura sobre los temas del módulo se encuentra dispersa, es extensa y posee acceso parcialmente limitado, el cuestionario de satisfacción sobre el material diseñado, obtuvo en todas las variables analizadas la categoría 4, lo que avala su actualización, pertinencia, integración, estructura y organización de los contenidos. Conclusiones: el material de apoyo resultó adecuado, con buena aceptación, y se tuvieron en cuenta las recomendaciones emitidas por los especialistas en su elaboración final.

ABSTRACT Background: the vertiginous development of science and technology constantly incorporates new advances to the daily task of medical practice, which calls for continuous training. This need is accentuated when dealing with the care of serious patients in the context of postoperative cardiovascular surgery, where the most of them are treated in intensive care units. Objectives: to design a teaching aid for bibliographic support for the subjects included in Module 25: Cardiovascular post-operative care in adults and children, of the third year study program of the cardiology residency. Methods: a development research was carried out in the "Ernesto Che Guevara" Cardiologic center, in Villa Clara, from December 2017 to 2018. Theoretical methods were used: analysis-synthesis, induction-deduction and systemic-structural; empirical ones: documentary analysis and questionnaire to specialized medical teachers; and statistical mathematics for absolute and relative values. Results: the literature on the subjects of the module is scattered, it is extensive and has partially limited access, the satisfaction questionnaire on the teaching aid designed, obtained in the entire variables analyzed category 4, which supports its updating, relevance, integration, structure and organization of the contents. Conclusions: the teaching aid was adequate, with good acceptance, and the recommendations issued by the specialists in its final elaboration were taken into account.

Diagnóstico bioquímico de acidurias orgánicas en Cuba: periodo 2008-2013 / Biochemical diagnosis of organic acidurias in Cuba: 2008-2013 period

Las acidurias orgánicas son las enfermedades metabólicas hereditarias más frecuentes en pacientes pediátricos con estado crítico de enfermedad. Este trabajo describe la experiencia cubana en el diagnóstico de estas enfermedades.Desde julio de 2008 hasta diciembre de 2013 se procesaron las orinas de pacientes cubanos con sospecha de aciduria orgánica. El perfil de ácidos orgánicos se obtuvo por cromatografía gaseosa acoplada a espectrometría de masas. Se analizaron muestras de orinas de 488 pacientes cubanos provenientes de todo el país. Del total de muestras procesadas, 29 mostraron niveles elevados de ácido láctico, pero no se realizó el diagnóstico confirmatorio de acidurias lácticas primarias en estos casos. Otras acidurias frecuentes entre los casos analizados fueron: acidurias metilmalónicas (5),deficiencia de acil-CoA deshidrogenasa de cadena media (3), alcaptonuria(3) y aciduria propiónica (3). Los hallazgos clínicos más frecuentes en los pacientes diagnosticados fueron la acidosis metabólica y los signos neurológicos. Los resultados muestran la importancia del análisis del perfil de ácidos orgánicos en niños con estado de gravedad; el diagnóstico acertado de estas enfermedades es decisivo para definir el tratamiento más efectivo y ofrecer a sus familias asesoramiento genético y la posibilidad de diagnóstico prenatal...

Performance of an in-house human immunodeficiency virus type 1 genotyping system for assessment of drug resistance in Cuba.

As commercial human immunodeficiency virus type 1 drug resistance assays are expensive, they are not commonly used in resource-limited settings. Hence, a more affordable in-house procedure was set up taking into account the specific epidemiological and economic circumstances of Cuba. The performance characteristics of the in-house assay were evaluated using clinical samples with various subtypes and resistance patterns. The lower limit of amplification was determined on dilutions series of 20 clinical isolates and ranged from 84 to 529 RNA copies/mL. For the assessment of trueness, 14 clinical samples were analyzed and the ViroSeq HIV-1 Genotyping System v2.0 was used as the reference standard. The mean nucleotide sequence identity between the two assays was 98.7% ± 1.0. Additionally, 99.0% of the amino acids at drug resistance positions were identical. The sensitivity and specificity in detecting drug resistance mutations was respectively 94.1% and 99.5%. Only few discordances in drug resistance interpretation patterns were observed. The repeatability and reproducibility were evaluated using 10 clinical samples with 3 replicates per sample. The in-house test was very precise as nucleotide sequence identity among paired nucleotide sequences ranged from 98.7% to 99.9%. The acceptance criteria were met by the in-house test for all performance characteristics, demonstrating a high degree of accuracy. Subsequently, the applicability in routine clinical practice was evaluated on 380 plasma samples. The amplification success rate was 91% and good quality consensus sequences encoding the entire protease and the first 335 codons in reverse transcriptase could be obtained for 99% of the successful amplicons. The reagent cost per sample using the in-house procedure was around € 80 per genotyping attempt. Overall, the in-house assay provided good results, was feasible with equipment and reagents available in Cuba and was half as expensive as commercial assays.

Safety and Immunogenicity of Cuban Antipneumococcal Conjugate Vaccine PCV7-TT in Healthy Adults.

INTRODUCTION Pneumococcal infections are a major cause of morbidity and mortality and are associated with considerable economic burden on health systems. To prevent pneumococcal infections, 7-valent conjugate vaccines have been available for over a decade; more recently, 10- and 13-valent conjugate vaccines have been formulated, which are more immunogenic than vaccines with capsular polysaccharides only. In Cuba, a new vaccine candidate has been developed, PCV7-TT, a conjugate of tetanus toxoid with antigens of seven of the serotypes of Streptococcus pneumoniae with highest circulation in Cuba and in the world: 1, 5, 6B, 14, 18C, 19F and 23F. OBJECTIVE Assess the safety of the vaccine candidate PCV7-TT in healthy adults and conduct a preliminary assessment of its immunogenicity. METHODS A phase I, double-blind clinical trial was performed at the National Toxicology Center in Havana, Cuba. Healthy male volunteers aged 18-35 years were randomly assigned to two groups: 20 received the vaccine candidate PCV7-TT and 20 the polyvalent antipneumococcal vaccine PNEUMO-23 used as control, each in a single intramuscular dose. To assess safety, the occurrence of adverse events was monitored for 30 days following inoculation. To explore immunogenicity, concentrations of serotype-specific antibodies was quantified before and 30 days after inoculation, as well titers of opsonophagocytic antibodies. (National Clinical Trial Registry RPCEC00000133) RESULTS Local adverse events were pain, redness, induration, increased sensitivity to touch, and warmth in the injection area. Pain was registered in 70% of individuals who received PCV7-TT and in 75% of those vaccinated with PNEUMO-23. Reported systemic adverse events were general malaise, headache and drowsiness. All adverse events appeared in the first 72 hours post inoculation and lasted no longer than 3 days. One event was reported that was classified as severe in intensity and serious in consequences, but it was unrelated to vaccination appendicitis in one individual inoculated with the control vaccine. Before vaccination, all participants but one had antibody concentrations =0.20 µg/ml against the vaccine strains; after vaccination 100% of individuals were positive and the concentrations of antibodies increased in previously positive volunteers. Some individuals had opsonophagocytic antibodies against serotypes 1, 14, 19F and 23F before vaccination, with highest concentrations against serotypes 14 and 19F. After vaccination, the percent of individuals with opsonophagocytic titers >1:8 for all serotypes in the vaccine was >50% in both groups. CONCLUSIONS A single dose of candidate vaccine PCV7-TT was safe when used in healthy adults. Preliminary results showed that it was able to activate an immune response against the serotypes of Streptococcus pneumoniae used. KEYWORDS Invasive pneumococcal diseases, pneumococcal vaccines, conjugate vaccines, immunization, randomized clinical trial, safety, Cuba.

Prospective, comprehensive, and effective viral monitoring in Cuban children undergoing solid organ transplantation.

PURPOSE: In Cuba, viral monitoring in the post-transplant period was not routinely performed. The aim of this research is to identify the most frequent viruses that affect transplanted Cuban children, by implementing a viral follow-up during the post-transplant period. METHODS: The study population included all Cuban pediatric patients who underwent solid organ transplantation (SOT) between November 2009 and December 2012. A total of 34 transplanted pediatric patients of kidney (n = 11) and liver (n = 23) were prospectively monitored during a 34-week period for viral DNAemia and DNAuria by simultaneous detection of cytomegalovirus (CMV), Epstein-Barr virus, herpes simplex virus type 1 and 2, varicella zoster virus, human herpesvirus 6, human adenovirus, and polyomaviruses (BKV and JCV) using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: Viral genome of at least one virus was detected in 21 of 34 recipients, 18 patients excreted virus in urine while 12 presented DNAemia. CMV (41.2%) and BKV (35.3%) were the most frequent viruses detected during the follow-up. CMV was the virus mainly associated with clinical symptoms and DNAemia. Its excretion in urine (with cut off value of 219 copies/mL) was associated with detection in plasma (p < 0.001); furthermore, CMV viruria was predictive of CMV viremia (OR:8.4, CI:2.4-29.1, p = 0.001). There was no association between high viral load and clinical complications, due to the prompt initiation of preemptive ganciclovir. CONCLUSION: This comprehensive viral monitoring program effectively prevents the development of critical viral disease, thus urge the implementation of qRT-PCR as routine for viral monitoring of transplanted Cuban organ recipients.

Molecular epidemiology of human papillomavirus infections in cervical samples from cuban women older than 30 years.

OBJECTIVE: This study aimed to provide information about the molecular epidemiology of human papillomavirus (HPV) in a group of Cuban women. MATERIALS AND METHODS: DNA from cervical samples was analyzed using a quantitative real-time polymerase chain reaction (PCR), which detects 6 of the clinically most relevant high-risk HPV types. Furthermore, end point PCR and sequencing were performed. Three hundred twenty-two women (211 with positive and 111 with negative cytologic results) aged between 30 and 69 years were enrolled. Risk factors associated with HPV infections and premalignant lesions were also investigated. RESULTS: HPV DNA was detected in 76.1% (245/322) of the studied population, and 34 different genotypes were found. There was an association between HPV infection and low educational level, history of oral contraceptives, menopausal stage, as well as cigarette and/or alcohol consumption. Besides, in a multivariate analysis, previous positive Pap test result and positive colposcopy finding were both predictor variables for HPV infections and for premalignant lesions. Human papillomavirus infection was found in 94.3% of women (199/211) with positive cytologic result and in 41.4% (46/111) of those with negative results, being more likely that the first group was infected with any HPV (odds ratio = 23.43; 95% CI = 11.70-46.92; p = .000). The most common genotypes were HPV types 16, 18, 31, 58, 33, and 45. All the cases with HPV positive findings had at least 1 high-risk HPV genotype. CONCLUSIONS: This is the first report of the molecular epidemiology of HPV in Cuban women, based on results from a DNA sequence and quantitative PCR. Most individuals were infected with high-risk HPV types. These findings support the inclusion of HPV vaccine in Cuba.

Antiretroviral drug resistance in HIV-1 therapy-naive patients in Cuba.

In Cuba, antiretroviral therapy rollout started in 2001 and antiretroviral therapy coverage has reached almost 40% since then. The objectives of this study were therefore to analyze subtype distribution, and level and patterns of drug resistance in therapy-naive HIV-1 patients. Four hundred and one plasma samples were collected from HIV-1 therapy-naive patients in 2003 and in 2007-2011. HIV-1 drug resistance genotyping was performed in the pol gene and drug resistance was interpreted according to the WHO surveillance drug-resistance mutations list, version 2009. Potential impact on first-line therapy response was estimated using genotypic drug resistance interpretation systems HIVdb version 6.2.0 and Rega version 8.0.2. Phylogenetic analysis was performed using Neighbor-Joining. The majority of patients were male (84.5%), men who have sex with men (78.1%) and from Havana City (73.6%). Subtype B was the most prevalent subtype (39.3%), followed by CRF20-23-24_BG (19.5%), CRF19_cpx (18.0%) and CRF18_cpx (10.3%). Overall, 29 patients (7.2%) had evidence of drug resistance, with 4.0% (CI 1.6%-4.8%) in 2003 versus 12.5% (CI 7.2%-14.5%) in 2007-2011. A significant increase in drug resistance was observed in recently HIV-1 diagnosed patients, i.e. 14.8% (CI 8.0%-17.0%) in 2007-2011 versus 3.8% (CI 0.9%-4.7%) in 2003 (OR 3.9, CI 1.5-17.0, p=0.02). The majority of drug resistance was restricted to a single drug class (75.8%), with 55.2% patients displaying nucleoside reverse transcriptase inhibitor (NRTI), 10.3% non-NRTI (NNRTI) and 10.3% protease inhibitor (PI) resistance mutations. Respectively, 20.7% and 3.4% patients carried viruses containing drug resistance mutations against NRTI+NNRTI and NRTI+NNRTI+PI. The first cases of resistance towards other drug classes than NRTI were only detected from 2008 onwards. The most frequent resistance mutations were T215Y/rev (44.8%), M41L (31.0%), M184V (17.2%) and K103N (13.8%). The median genotypic susceptibility score for the commonly prescribed first-line therapies was 2.5. This analysis emphasizes the need to perform additional surveillance studies to accurately assess the level of transmitted drug resistance in Cuba, as the extent of drug resistance might jeopardize effectiveness of first-line regimens prescribed in Cuba and might necessitate the implementation of baseline drug resistance testing.

High frequency of antiviral drug resistance and non-B subtypes in HIV-1 patients failing antiviral therapy in Cuba.

BACKGROUND: Emergence of HIV-1 drug resistance may limit the sustained benefits of antiretroviral therapy (ART) in settings with limited laboratory monitoring and drug options. OBJECTIVES: Surveillance of drug resistance and subtypes in HIV-1 patients failing ART in Cuba. STUDY DESIGN: This study compiled data of ART-experienced HIV-1 patients attending a clinical center in Havana in 2003 and 2009-2011. The first period included results of a cross-sectional study, whereas in the second period genotyping was performed as part of routine care. Drug resistance mutations and levels were determined using HIVdb version 6.0.9. RESULTS: Seventy-six percent received solely ART containing at least 3 drugs, of which 79.1% ever receiving unboosted protease inhibitors (PI). Patients from 2009 to 2011 were longer treated and exposed to more ART regimens. Subtype B (39%) and CRF19_cpx (18%) were the most prevalent genetic forms. Subtype distribution did not change significantly between both periods, except for BG recombinants that increased from 6% to 14%. Nucleoside reverse transcriptase inhibitor (NRTI), non-nucleoside RTI (NNRTI) and PI mutations were present in 69.5%, 54.8% and 44.4%. Full-class resistance (FCR) to NRTI, NNRTI, PI and multidrug resistance (MDR) were detected in 31.8%, 37.9%, 18.5% and 15.4%. FCR to NRTI, NNRTI, PI and MDR were present in 9.8%, 14.1%, 0%, 0% after first-line failure and in 19.8%, 20.8%, 2.9% and 2.9% after second-line failure. CONCLUSIONS: Our study found a high prevalence of drug resistance and supports the need for appropriate laboratory monitoring in clinical practice and access to drug options in case of virological failure.

Kaposi's Sarcoma and Human Herpesvirus 8 in Cuba: evidence of subtype B expansion.

OBJECTIVE: To evaluate the temporal distribution (1991-2009) and associated variation of KSHV subtypes in Cuba. METHOD: Phylogenetic characterization based on the KSHV K1 gene was performed using 90 KSHV positive samples. RESULTS: Molecular characterization confirmed the prevalence of a wide range of KSHV subtypes (A: n=48 [A5=12]; C: n=15; B: n=22; and E: n=5). In the current study, we observed a significant increase in HHV-8 subtype B after 2004 (p=0.0063). This Subtype B in Cuba was associated with: heterosexual behaviour (OR: 3.63, CI: 1,2-10,98; p=0.03), with the antecedent of acquiring HIV/KSHV in Africa (p=0.0003), with nodular stage of KS lesions (OR 4.2, CI: 1.1 to 15.7; p=0.04). CONCLUSION: Our study is the first to report KSHV Subtype B expansion in Cuba, that might be reflective of a change in human behavioural pattern.

Single-center clinical comparison of two reinforced ureteral access sheaths for retrograde ureteroscopic treatment of urinary lithiasis.

BACKGROUND: Ureteral access sheaths (UAS) are used to facilitate ureteroscopic procedures. Difficulties in use, including sheath distortion, buckling, and difficulty in placement, have been reported. However, few clinical comparisons have been performed. We present the first large-scale comparison of the Applied ForteXE(®) and ACMI-Gyrus Uropass(®) UAS. METHODS: We retrospectively compared patients who underwent ureteroscopy for urolithiasis with one of two types of UAS: Applied Forte XE(®) or ACMI-Gyrus Uropass(®). Demographics, operative parameters, and outcomes were assessed. Statistical analysis was performed. RESULTS: In 125 (64.4%) male and 69 (35.6%) female patients, 194 UAS were used. One hundred and thirteen (58.2%) Applied Forte XE(®) and 81 (41.8%) ACMI-Gyrus Uropass(®) were utilized. Success rates for sheath deployment were as follows: overall = 186/194 (95.8%); Applied Forte XE(®) = 107/113 (94.7%); and ACMI-Gyrus Uropass(®) = 79/81 (97.5%) (P = 0.472). Of the 194 patients 131 (67.5%) had a pre-existing stent. Sheath deployment failures occurred in 7 men and 1 woman, of which 4/8 (50%) had no pre-existing stent. Limitations of deployed sheaths occurred at low frequency in both Applied Forte XE 17/107 (15.9%) and ACMI-Gyrus 6/79 (7.6%), with no significant difference observed (P = 0.120). Limitations in use was high in men (P = 0.019). At a mean follow-up of 41 months, no ureteral strictures were noted. CONCLUSIONS: No significant differences were seen in overall success rates for both sheaths. Both sheaths had high deployment success rates and a similar low frequency of sheath-related limitations. We noted increased limitations in the use of deployed sheaths in men. Successful sheath use may depend on both the sheath itself and patient/operative parameters.

Assessment of infection with polyomaviruses BKV, JCV and SV40 in different groups of Cuban individuals.

We investigated the frequency of BKV, JCV and SV40 reactivation in three groups of Cuban patients by multiplex nested PCR assay of 40 paraffin-embedded colorectal neoplasm tissues, 113 urine samples, and 125 plasma samples from 27 transplant recipients, and cerebrospinal fluid (CSF) from 67 HIV-1-infected individuals with central nervous system (CNS) disorders. None of these polyomaviruses were detected in colorectal neoplasms. JCV DNA was detected in 2 of 67 patients (2.9%) with CNS disorders, but neither BKV nor SV40 was identified. BKV was found in urine from 38.5% and 28.6% of adult and pediatric transplant recipients, respectively. In adult renal transplant recipients, excretion of BKV in urine was significantly associated with episodes of acute rejection (p=0.012) and with excretion of HCMV in urine (p= 0.008). In Cuba, the polyomaviruses studied here could not be related to colorectal neoplasms, and JCV was rarely detected in CSFs of HIV-1-infected individuals, whilst BKV reactivation was found to occur frequently in organ transplant recipients.

Role of herpesvirus as a cause of mononucleosis-like and febrile syndromes in Cuba, 2006-2009.

INTRODUCTION: Herpesvirus infections are prevalent worldwide, but most run their course asymptomatically. Clinical presentations in symptomatic cases vary widely and include febrile and mononucleosis-like syndromes. In immunocompromised patients, herpetic infection can be lethal and routine laboratory tests are of little use. Use of novel techniques may provide important improvements in diagnosis and treatment of these patients. OBJECTIVE: Investigate association between different herpesviruses and the etiology of mononucleosis and febrile syndromes in Cuban immunocompetent and immunocompromised patients. METHODS: The study used multiplex nested polymerase chain reaction, enabling simultaneous detection of six herpesviruses--cytomegalovirus, herpes simplex (1 and 2), Epstein-Barr, varicella-zoster and human herpesvirus 6--to study 1157 samples (770 urine and 387 serum samples) from 1140 patients with mononucleosis-like syndrome or febrile syndrome, classified according to history of immunosuppressive disease. Samples were analyzed at the Laboratory for Sexually Transmitted Diseases (Virology) of the Pedro Kourí Tropical Medicine Institute from January 2006 through December 2009. SPSS statistical package was used and incidence rates calculated. RESULTS: Of samples studied, 20.1% were positive for some herpesvirus. Higher risk of developing active herpesvirus infections was detected in samples from immunocompromised patients with febrile syndrome compared to those of immunocompetent ones (OR 2.02, CI 1.20-3.42, p=0.007). Cytomegalovirus was the most frequently found herpesvirus in both mononucleosis-like syndrome (60.4%) and febrile syndrome (63.6%) and in both children (69.2%) and adults (55.2%), followed by Epstein-Barr virus. Cytomegalovirus was detected in 68.9% of positive urine samples and in just 47.2% of serum samples. CONCLUSIONS: This is the first Cuban study demonstrating the pathogenic role of herpesviruses, particularly cytomegalovirus, in patients with febrile or mononucleosis-like syndrome, in both immunocompetent and immunocompromised patients. Results highlight the importance of including molecular diagnosis of the herpesvirus family in investigating mononucleosis and febrile syndromes of unknown etiology and demonstrate that etiologic diagnosis would not have been feasible in many cases without the use of this diagnostic tool.

Mangifera indica L. extract (Vimang) and mangiferin reduce the airway inflammation and Th2 cytokines in murine model of allergic asthma.

OBJECTIVES: The aim was to study the effects of Mangifera indica extract and its major component mangiferin on lung inflammation response and Th2 cytokine production using a murine experimental model of allergic asthma. METHODS: BALB/c mice were intraperitoneally sensitized with 10 µg of ovoalbumin (OVA) adsorbed on aluminium hydroxide on days 0, 7 and 14. Seven days after the last injection, the mice were challenged with 2% aerosolized OVA inhalation for 30 min beginning on day 21 and continuing until day 24. To evaluate the protective effect, mice were orally treated with M. indica extract (50, 100 or 250 mg/kg) or mangiferin (50 mg/kg) from days 0 to 24. Anti-OVA immunoglobulin E, interleukin (IL)-4 and IL-5 were determined by ELISA and lungs were analysed by histology. KEY FINDINGS: M. indica extract and mangiferin produced a marked reduction of airway inflammation around vessels and bronchi, inhibition of IL-4 and IL-5 cytokines in bronchoalveolar lavage fluid and lymphocyte culture supernatant, IgE levels and lymphocyte proliferation. CONCLUSION: This is the first pre-clinical report of the anti-inflammatory properties of M. indica extract and mangiferin in experimental asthma and it could be an important part of pre-clinical requirement necessary for its use to complement the treatment of this complex disease.

Drug-resistant HIV-1 in Cuban children and their seropositive mothers.

INTRODUCTION The use of highly active antiretroviral therapy has reduced progression to AIDS and increased survival among seropositive persons; yet, appearance of resistant viruses may jeopardize these benefits. In Cuba, HIV mainly affects adults; at the end of 2009 of the 41 children infected, 25 were still alive; of these, 22 were under antiretroviral treatment. Until now, nothing was known about HIV-1 antiviral resistance and viral subtypes in the pediatric population in Cuba. OBJECTIVE This study aims to identify presence of antiretroviral-resistant HIV-1 strains in Cuban children and their mothers, and to provide a phylogenetic characterization and comparison of pol gene sequences in the same. METHODS Plasma samples were collected from 22 children and their mothers, all HIV-1-infected, from 2004 through 2009. Reverse transcription polymerase chain reaction was used to amplify the pol gene fragment coding for HIV protease and reverse transcriptase enzymes; this was then sequenced and subjected to phylogenetic analysis of HIV subtypes and recombinant forms to compare sequences between mothers and children. HIV mutations conferring antiretroviral resistance were determined. RESULTS Viral amplification was achieved in samples from 11 children and 8 mothers. Subtypes detected were: CRF19_cpx in five children, subtype B in three, CRF18_cpx in two, and subtype C in one child. In all mother-child pairs, samples were grouped within the same viral subtype in the phylogenetic tree. One mother was under treatment and five children had been treated before the sample was collected. In viruses amplified from samples of children under treatment, resistance was most frequently found to lamivudine (3 cases) and nevirapine (4 cases). Two untreated children carried resistant viruses possibly acquired from their mothers. CONCLUSIONS This is the first study to describe HIV-1 antiviral resistance in the pediatric population in Cuba; it also identified viral subtypes infecting the mother-child pairs studied. We recommend antiretroviral resistance assays before initiating treatment in pregnant seropositive women and their newborns.

Fibrilación auricular en el post quirúrgico dependiente de la hipertensión arterial como factor de riesgo pre operatorio / Fibrilação atrial no pós-cirúrgico relacionada à hipertensão arterial como fator de risco pré-operatório / Atrial fibrillation in the postoperative depending on the hypertension as a risk factor

Introducción: Las personas hipertensas tienen más probabilidades de presentar fibrilaciónauricular que los normotensos; de los pacientes con fibrilación auricular, el 57% son hipertensos. La fisiopatología de la fibrilación auricular post operatoria no está completamente clara, considerándose multifactorial. Objetivo: determinar los factores de riesgo prequirúrgicos que se asocian a la fibrilaciónauricular en el postoperatorio de la cirugía de revascularización miocárdica sin circulación extracorpórea.Material y Métodos: Se realizó un estudio retrospectivo en pacientes sometidos a cirugía revascularizadorasin circulación extracorpórea en el Cardiocentro de Santa Clara. Se estudiaron 110 pacientes entre eneroy diciembre del 2007. Los datos tales como edad, sexo, factores de riesgo, antecedentes patológicospersonales y uso de medicamentos antes de la cirugía, fueron recogidos. Resultados: En análisismultivariado de regresión logística, la hipertensión arterial es el único, predictor independiente para desarrollarfibrilación auricular (OR 4.56; IC 0.96-21.69; p 0.050). Discusión: Hay evidencia que indica que la disfuncióndiastólica, así como el grado de hipertrofia ventricular izquierda, del paciente hipertenso aumenta el riesgode fibrilación auricular. La fibrilación auricular es una arritmia eléctricamente benigna pero su aparición puedeconducir a complicaciones neurológicas y deterioro hemodinámico. Conclusiones: Los únicos factoresinfluyentes en el desarrollo de la fibrilación auricular en el postquirúrgico no son los factores de riesgoprequirúrgicos, esta es multifactorial, pero el control de estos que se asocian a la fibrilación auricular enel postoperatorio de la cirugía de revascularización miocárdica, disminuiría el riesgo de arritmia.