Identificación del alelo HLA B*57: 01 en una población militar, de Lima-Perú / Identification of the allele HLA B*57: 01 in a military population, from Lima-Peru

El alelo HLA B*57:01 es un marcador genético asociado con la hipersensibilidad al fármaco anti-retroviral abacavir (ABC) y su frecuencia en la población peruana todavía es desconocida. El objetivo fue identificar el alelo HLA B*57:01 en una población militar de Lima, Perú. Se reclutaron 43 personas viviendo con VIH (PVV) quienes aceptaron participar a través de un consentimiento informado. La detección del alelo HLA B*57:01 se realizó mediante RPC en tiempo real (RT-PCR). Asimismo, se determinó la carga viral (CV), el recuento de linfocitos CD4 y la genotipificación del VIH. Se identificaron dos casos positivos al alelo HLA B*57:01 (4,7%). Además, uno de ellos presentó múltiples mutaciones de resistencia a los anti-retrovirales (ARV), incluyendo ABC. Se demostró por primera vez en el Perú la presencia del alelo HLA B*57:01.

The HLA B*57:01 allele is a genetic marker associated with hypersensitivity to the antiretroviral Abacavir (ABC) and its frequency in the Peruvian population is still unknown. The objective was to identify the HLA B*57:01 allele in a military population from Lima, Peru. Forty three people living with HIV (PLWH) were recruited, who agreed to participate through informed consent. Detection of the HLA B*57:01 allele was performed by real-time PCR (RT-PCR). Likewise, viral load (VL), CD4 lymphocyte count and HIV genotyping were determined. Two cases positive for the HLA B*57:01 allele (4.7%) were identified. In addition, one of them had multiple resistance mutations to antiretrovirals (ARVs), including ABC. The presence of the HLA B*57:01 allele was demonstrated for the first time in Peru.

Antibacterial and Antiviral Properties of Chenopodin-Derived Synthetic Peptides.

Antimicrobial peptides have been developed based on plant-derived molecular scaffolds for the treatment of infectious diseases. Chenopodin is an abundant seed storage protein in quinoa, an Andean plant with high nutritional and therapeutic properties. Here, we used computer- and physicochemical-based strategies and designed four peptides derived from the primary structure of Chenopodin. Two peptides reproduce natural fragments of 14 amino acids from Chenopodin, named Chen1 and Chen2, and two engineered peptides of the same length were designed based on the Chen1 sequence. The two amino acids of Chen1 containing amide side chains were replaced by arginine (ChenR) or tryptophan (ChenW) to generate engineered cationic and hydrophobic peptides. The evaluation of these 14-mer peptides on Staphylococcus aureus and Escherichia coli showed that Chen1 does not have antibacterial activity up to 512 µM against these strains, while other peptides exhibited antibacterial effects at lower concentrations. The chemical substitutions of glutamine and asparagine by amino acids with cationic or aromatic side chains significantly favoured their antibacterial effects. These peptides did not show significant hemolytic activity. The fluorescence microscopy analysis highlighted the membranolytic nature of Chenopodin-derived peptides. Using molecular dynamic simulations, we found that a pore is formed when multiple peptides are assembled in the membrane. Whereas, some of them form secondary structures when interacting with the membrane, allowing water translocations during the simulations. Finally, Chen2 and ChenR significantly reduced SARS-CoV-2 infection. These findings demonstrate that Chenopodin is a highly useful template for the design, engineering, and manufacturing of non-toxic, antibacterial, and antiviral peptides.

Consentimiento informado y directivas anticipadas: análisis comparado de la legislación en América Latina / Consentiment informat i directives anticipades: anàlisi comparada de la legislació a Amèrica Llatina / Informed consent and living wills: comparative analysis of the legislation in Latin America

Objetivos: Describir la forma en que se establecen normativamente el consentimiento informado (CI) y las directivas anticipadas (DA) en países latinoamericanos en relación a la asistencia en salud y la investigación, y su análisis desde la bioética. Métodos: Se realizó un análisis comparativo de las normativas vigentes de los 21 países latinoamericanos respecto al CI y las DA. Se confeccionaron reportes de extracción de datos que fueron diseñados antes de iniciar el proceso de investigación para cada país. Se analizaron cualitativamente similitudes y diferencias respecto a variables predefinidas. Se realizaron tablas para sintetizar la información donde fue posible.Resultados:Dieciocho países latinoamericanos prevén una norma específica para el uso del CI en el ámbito clínico. Seis países han incorporado normativas respecto al uso del CI en la virtualidad. Respecto al uso del CI en investigación, 20 países latinoamericanos prevén una norma específica para este ámbito. En el caso de la exposición académica, son 6 países los que nombran esta situación en la normativa. Siete países latinoamericanos cuentan con normativa para establecer DA.Conclusiones:América Latina ha presentado una notable evolución en torno a normar dos de los más importantes instrumentos del ejercicio de la autonomía, como los son el CI y las DA. La legislación sobre ambos instrumentos no garantiza mayor autonomía en salud, pero su existencia como reguladora de acciones para la toma de decisiones constituye el primer paso para su ejercicio.(AU)

Objectius: Descriure la manera com s'estableixen normativament el consentiment informat (CI) i les directives anticipades (DA) en països llatinoamericans en relació amb l'assistència en salut i la investigació, i la seva anàlisi des de la bioètica. Mètodes: Es va realitzar una anàlisi comparativa de les normatives vigents dels 21 països llatinoamericans respecte al CI i les DA. Es van confeccionar reports d’extracció de dades que van ser dissenyats abans d’iniciar el procés de recerca per a cada país. Es van analitzar qualitativament similituds i diferències respecte de variables predefinides. Es van fer taules per sintetitzar la informació on va ser possible.Resultats:Divuit països llatinoamericans preveuen una norma específica per a l'ús del CI a l'àmbit clínic. Sis països han incorporat normatives respecte a l'ús del CI a la virtualitat. Pel que fa a l'ús del CI eninvestigació, 20 països llatinoamericans preveuen una norma específica per a aquest àmbit. En el cas de l'exposició acadèmica, són 6 països els que nomenen aquesta situació a la normativa. Set països llatinoamericans tenen normativa per establir DA.Conclusions:Amèrica Llatina ha presentat una notable evolució al voltant de normar dos dels instruments més importants de l'exercici de l'autonomia, com els són el CI i les DA. La legislació sobre tots dos instruments no garanteix més autonomia en salut, però la seva existència com a reguladora d'accions per a la presa de decisions constitueix el primer pas per al seu exercici.(AU)

Objectives: To describe how informed consent (IC) and living wills (LW) are legally established in Latin American countries in relation to health care and research, and to analyse that from bioethics. Methods: A comparative analysis of the current regulations of the 21 Latin American countries regarding CI and LW was carried out. Data extraction reports were prepared before starting the research process for each country. Similarities and differences were qualitatively analyzed according to predefined variables. Tables were made to synthesize the information where possible.Results:Eighteen Latin American countries provide a specific criteria for IC use in clinical setting. Six countries have incorporated regulations regarding the use of ICin virtually environment. Regarding the use of IC in research, 20 Latin American countries provide a specific regulation. In the case of the academic exposure, 6 countries name this situation in their regulations. Seven Latin American countries have regulations to establish LW.Conclusions:Latin America has presented a notable evolution around regulating two of the most important instruments of the exercise of autonomy, such as the CI and the DA. The legislation on both instruments does not guarantee that there is greater autonomy in health, but its existence as a regulator of actions for decision-making constitutes the first step for its exercise.(AU)

New Drug Design Avenues Targeting Alzheimer's Disease by Pharmacoinformatics-Aided Tools.

Neurodegenerative diseases (NDD) have been of great interest to scientists for a long time due to their multifactorial character. Among these pathologies, Alzheimer's disease (AD) is of special relevance, and despite the existence of approved drugs for its treatment, there is still no efficient pharmacological therapy to stop, slow, or repair neurodegeneration. Existing drugs have certain disadvantages, such as lack of efficacy and side effects. Therefore, there is a real need to discover new drugs that can deal with this problem. However, as AD is multifactorial in nature with so many physiological pathways involved, the most effective approach to modulate more than one of them in a relevant manner and without undesirable consequences is through polypharmacology. In this field, there has been significant progress in recent years in terms of pharmacoinformatics tools that allow the discovery of bioactive molecules with polypharmacological profiles without the need to spend a long time and excessive resources on complex experimental designs, making the drug design and development pipeline more efficient. In this review, we present from different perspectives how pharmacoinformatics tools can be useful when drug design programs are designed to tackle complex diseases such as AD, highlighting essential concepts, showing the relevance of artificial intelligence and new trends, as well as different databases and software with their main results, emphasizing the importance of coupling wet and dry approaches in drug design and development processes.

Impacto en la salud mental en residentes de tres programas de especialización médica de la Universidad de Valparaíso durante la pandemia por COVID-19 / Impact of the COVID-19 pandemic on the mental health of residents of three specialty medical programs

BACKGROUND: COVID-19 pandemic disturbed mental health of healthcare personnel. Residents of the specialization programs could be at risk, since they were reassigned in their functions. Aim: To describe the impact of COVID-19 pandemic on symptoms of depression, stress, anxiety and resilient coping in residents of Anesthesiology, Internal Medicine and Emergency Medicine MATERIAL AND METHODS: Residents were invited to answer an online survey containing the DASS-21 scale for anxiety, stress and depression symptoms and the Brief Resilient Coping Scale (BRCS) for resilience skills. RESULTS: Fifty four out of 90 residents answered the survey. Eighteen to 24% of respondents had symptoms of depression, anxiety and stress at severe and extremely severe levels. Those with severe and extremely severe symptoms had also the lowest score on the BRCS resilience scale. We did not find an association between severity of symptoms and gender. Discussion: A proportion of respondent residents had severe psychological symptoms and lower resilience scores during the COVID-19 pandemic.

Structural Insights into the Substrate Transport Mechanisms in GTR Transporters through Ensemble Docking.

Glucosinolate transporters (GTRs) are part of the nitrate/peptide transporter (NPF) family, members of which also transport specialized secondary metabolites as substrates. Glucosinolates are defense compounds derived from amino acids. We selected 4-methylthiobutyl (4MTB) and indol-3-ylmethyl (I3M) glucosinolates to study how GTR1 from Arabidopsis thaliana transports these substrates in computational simulation approaches. The designed pipeline reported here includes massive docking of 4MTB and I3M in an ensemble of GTR1 conformations (in both inward and outward conformations) extracted from molecular dynamics simulations, followed by clustered and substrate-protein interactions profiling. The identified key residues were mutated, and their role in substrate transport was tested. We were able to identify key residues that integrate a major binding site of these substrates, which is critical for transport activity. In silico approaches employed here represent a breakthrough in the plant transportomics field, as the identification of key residues usually takes a long time if performed from a purely wet-lab experimental perspective. The inclusion of structural bioinformatics in the analyses of plant transporters significantly speeds up the knowledge-gaining process and optimizes valuable time and resources.

[Impact of the COVID-19 pandemic on the mental health of residents of three specialty medical programs]. / Impacto en la salud mental en residentes de tres programas de especialización médica de la Universidad de Valparaíso durante la pandemia por COVID-19.

BACKGROUND: COVID-19 pandemic disturbed mental health of healthcare personnel. Residents of the specialization programs could be at risk, since they were reassigned in their functions. AIM: To describe the impact of COVID-19 pandemic on symptoms of depression, stress, anxiety and resilient coping in residents of Anesthesiology, Internal Medicine and Emergency Medicine Material and Methods: Residents were invited to answer an online survey containing the DASS-21 scale for anxiety, stress and depression symptoms and the Brief Resilient Coping Scale (BRCS) for resilience skills. RESULTS: Fifty four out of 90 residents answered the survey. Eighteen to 24% of respondents had symptoms of depression, anxiety and stress at severe and extremely severe levels. Those with severe and extremely severe symptoms had also the lowest score on the BRCS resilience scale. We did not find an association between severity of symptoms and gender. DISCUSSION: A proportion of respondent residents had severe psychological symptoms and lower resilience scores during the COVID-19 pandemic.

Application of ensemble pharmacophore-based virtual screening to the discovery of novel antimitotic tubulin inhibitors.

Tubulin is a well-validated target for herbicides, fungicides, anti-parasitic, and anti-tumor drugs. Many of the non-cancer tubulin drugs bind to its colchicine site but no colchicine-site anticancer drug is available. The colchicine site is composed of three interconnected sub-pockets that fit their ligands and modify others' preference, making the design of molecular hybrids (that bind to more than one sub-pocket) a difficult task. Taking advantage of the more than eighty published X-ray structures of tubulin in complex with ligands bound to the colchicine site, we generated an ensemble of pharmacophore representations that flexibly sample the interactional space between the ligands and target. We searched the ZINC database for scaffolds able to fit several of the subpockets, such as tetrazoles, sulfonamides and diarylmethanes, selected roughly ~8000 compounds with favorable predicted properties. A Flexi-pharma virtual screening, based on ensemble pharmacophore, was performed by two different methodologies. Combining the scaffolds that best fit the ensemble pharmacophore-representation, we designed a new family of ligands, resulting in a novel tubulin modulator. We synthesized tetrazole 5 and tested it as a tubulin inhibitor in vitro. In good agreement with the design principles, it demonstrated micromolar activity against in vitro tubulin polymerization and nanomolar anti-proliferative effect against human epithelioid carcinoma HeLa cells through microtubule disruption, as shown by immunofluorescence confocal microscopy. The integrative methodology succedes in the design of new scaffolds for flexible proteins with structural coupling between pockets, thus expanding the way in which computational methods can be used as significant tools in the drug design process.

Antimicrobial properties of novel ionic liquids derived from imidazolium cation with phenolic functional groups.

Bacterial infections are nowadays among the major threats to public health worldwide. Thus, there is an urgent and increased need for new antimicrobial agents. As a result, the exploration of the antimicrobial properties of different substances including ionic liquids (ILs) has recently attracted great attention. The present work is aimed at evaluating how the addition of halogens and hydrophobic substituents on alkylimidazolium units of ILs as well as the increase in their chain lengths affects the antimicrobial properties of such ILs. After their synthesis, the antibacterial activities of these compounds against Pseudomona aeruginosa, Escherichia coli, and Staphylococcus aureus are determined by measuring their minimal inhibitory concentrations (MICs). Key features in ILs-membrane interactions are also studied using long-term all-atom molecular dynamics simulations (MDs). The results show that these ILs have good antibacterial activity against S. aureus, E. coli, and P. aeruginosa, with MIC values range from <7.81 to 62.50 µM. The antimicrobial property of tert-butyl N-methylphenolimidazolium salts (denoted as 8b and 8c) is particularly better with MIC values of < 7.81 µM. The antibacterial efficacy is also found to depend on the alkyl chain length and substituents on the phenolic ring. Finally, MDs done for ILs in a phosphatidylcholine (POPC) bilayer show key features in the mechanism of IL-induced membrane disruption, where the ILs are inserted as clusters into one side of the bilayer until saturation is reached. This insertion increases "leaflet strain" up to critical threshold, likely triggering the morphological disruption of the membranes in the microbes.

PSC-db: A Structured and Searchable 3D-Database for Plant Secondary Compounds.

Plants synthesize a large number of natural products, many of which are bioactive and have practical values as well as commercial potential. To explore this vast structural diversity, we present PSC-db, a unique plant metabolite database aimed to categorize the diverse phytochemical space by providing 3D-structural information along with physicochemical and pharmaceutical properties of the most relevant natural products. PSC-db may be utilized, for example, in qualitative estimation of biological activities (Quantitative Structure-Activity Relationship, QSAR) or massive docking campaigns to identify new bioactive compounds, as well as potential binding sites in target proteins. PSC-db has been implemented using the open-source PostgreSQL database platform where all compounds with their complementary and calculated information (classification, redundant names, unique IDs, physicochemical properties, etc.) were hierarchically organized. The source organism for each compound, as well as its biological activities against protein targets, cell lines and different organism were also included. PSC-db is freely available for public use and is hosted at the Universidad de Talca.

Analysis of the Zonula occludens Toxin Found in the Genome of the Chilean Non-toxigenic Vibrio parahaemolyticus Strain PMC53.7.

Vibrio parahaemolyticus non-toxigenic strains are responsible for about 10% of acute gastroenteritis associated with this species, suggesting they harbor unique virulence factors. Zonula occludens toxin (Zot), firstly described in Vibrio cholerae, is a secreted toxin that increases intestinal permeability. Recently, we identified Zot-encoding genes in the genomes of highly cytotoxic Chilean V. parahaemolyticus strains, including the non-toxigenic clinical strain PMC53.7. To gain insights into a possible role of Zot in V. parahaemolyticus, we analyzed whether it could be responsible for cytotoxicity. However, we observed a barely positive correlation between Caco-2 cell membrane damage and Zot mRNA expression during PMC53.7 infection and non-cytotoxicity induction in response to purified PMC53.7-Zot. Unusually, we observed a particular actin disturbance on cells infected with PMC53.7. Based on this observation, we decided to compare the sequence of PMC53.7-Zot with Zot of human pathogenic species such as V. cholerae, Campylobacter concisus, Neisseria meningitidis, and other V. parahaemolyticus strains, using computational tools. The PMC53.7-Zot was compared with other toxins and identified as an endotoxin with conserved motifs in the N-terminus and a variable C-terminal region and without FCIGRL peptide. Notably, the C-terminal diversity among Zots meant that not all of them could be identified as toxins. Structurally, PMC53.7-Zot was modeled as a transmembrane protein. Our results suggested that it has partial 3D structure similarity with V. cholerae-Zot. Probably, the PMC53.7-Zot would affect the actin cytoskeletal, but, in the absence of FCIGRL, the mechanisms of actions must be elucidated.

The subjective experience of psychotherapists during moments of rupture in psychotherapy with adolescents.

The study of the ruptures of the therapeutic alliance has impacted research in psychotherapy by highlighting the relational nature of this phenomenon. Despite ruptures are frequent and relevant during adolescent psychotherapy, most of the empirical evidence in this field has been carried out with adults. Understanding the subjective experience of the therapist during ruptures while working with adolescent is proposed as a starting point for the study of this type of interactional scenarios. The study examined the meanings that emerge from the therapists' experience in terms of their explanations about the causes and effects of ruptures with adolescents. Eight psychotherapists were interviewed about their experiences during ruptures with young patients. The data was qualitatively analyzed through the Interpretive Phenomenological Analysis method. Four categories emerged: the failure to recognize the adolescent's experience, the intensity of the affective experience of adolescents in psychotherapy, therapeutic boundaries as an articulator of the therapeutic purpose and, the obstacles that family generates during the therapeutic process. This study concurs with the literature on the need to make explicit with the family about the meaning, roles and limits of the therapy, and to prevent the exercise of control from an adultcentered position. It is concluded that in order to avoid and repair ruptures with adolescents in psychotherapy, an approach that integrates a sensitive attitude, an ecological point of view and mentalizing about the origin of the rupture is needed.

Manual de procedimientos de electroforesis para proteínas y ADN / Procedures Manual for protein and DNA electrophoresis

El presente manual de procedimientos es la recopilación de protocolos actualizados por los investigadores de la División de Biología Molecular del Instituto Nacional de Salud. En este material se incluye, además de los procedimientos técnicos de la electroforesis, un anexo indicando el modo de preparación de soluciones de electroforesis, unidades y fórmulas básicas a ser aplicadas y algunas importantes medidas de bioseguridad que deben considerarse durante la manipulación de los reactivos. Es importante señalar que la electroforesis en asociación con otras técnicas tales como PCR, e hibridación, brinda una gran ayuda en el campo de la salud. Gracias a esta técnica es posible visualizar las bandas de ARN o ADN de patógenos, detectar cambios o mutaciones a nivel genético, visualizar una nueva proteína, entre otros

Manual de procedimientos de electroforesis para proteínas y ADN / Procedures Manual for protein and DNA electrophoresis

El presente manual de procedimientos es la recopilación de protocolos actualizados por los investigadores de la División de Biología Molecular del Instituto Nacional de Salud. En este material se incluye, además de los procedimientos técnicos de la electroforesis, un anexo indicando el modo de preparación de soluciones de electroforesis, unidades y fórmulas básicas a ser aplicadas y algunas importantes medidas de bioseguridad que deben considerarse durante la manipulación de los reactivos. Es importante señalar que la electroforesis en asociación con otras técnicas tales como PCR, e hibridación, brinda una gran ayuda en el campo de la salud. Gracias a esta técnica es posible visualizar las bandas de ARN o ADN de patógenos, detectar cambios o mutaciones a nivel genético, visualizar una nueva proteína, entre otros(AU)

Caracterización Molecular de la Secuencia parcialdel Gen de la Glicoproteína NS1 del Virus Dengue 1 proveniente de Máncora, Perú

Se caracterizó una región genética que codifica la glicoproteína NS1 del virus dengue 1 proveniente de Máncora, Piura. Comparaciones de secuencias de nucleótidos revelaron un 93,32% de identidad entre el aislamiento peruano y una cepa de Hawai. A nivel de aminoácidos, se observaron cambios de tipo no conservativos en dominios epitópicos de reconocimiento humoral. De otro lado, el perfil hidropático de la región estudiada fue similar al de otros aislamientos referenciales. Los resultados sugieren realizar mayores análisis de identidad genética y mutaciones en dominios epitópicos en el virus dengue 1 peruano.

A 419bp-NS1 genetic region corresponding to Dengue virus 1 was characterised from an outbreak in Máncora Piura. The comparison of nucleotide sequences revealed that Peruvian isolates showed high correlation (93.32%) with a Hawaii strain. Amino acids comparisons revealed non-conservative changes into humoral response epitope domains. On the other hand, the hydropathy profile of NS1 was similar to other referential strains. The results suggest that more comparisons are needed regarding genetic identity and mutations into the epitope domain of Peruvian dengue 1 virus.