Comparative study of SARS-CoV-2 infection in different cell types: Biophysical-computational approach to the role of potential receptors.

Cellular susceptibility to SARS-CoV-2 infection in the respiratory tract has been associated with the ability of the virus to interact with potential receptors on the host membrane. We have modeled viral dynamics by simulating various cellular systems and artificial conditions, including macromolecular crowding, based on experimental and transcriptomic data to infer parameters associated with viral growth and predict cell susceptibility. We have accomplished this based on the type, number and level of expression of the angiotensin-converting enzyme 2 (ACE2), transmembrane serine 2 (TMPRSS2), basigin2 (CD147), FURIN protease, neuropilin 1 (NRP1) or other less studied candidate receptors such as heat shock protein A5 (HSPA5) and angiotensin II receptor type 2 (AGTR2). In parallel, we studied the effect of simulated artificial environments on the accessibility to said proposed receptors. In addition, viral kinetic behavior dependent on the degree of cellular susceptibility was predicted. The latter was observed to be more influenced by the type of proteins and expression level, than by the number of potential proteins associated with the SARS CoV-2 infection. We predict a greater theoretical propensity to susceptibility in cell lines such as NTERA-2, SCLC-21H, HepG2 and Vero6, and a lower theoretical propensity in lines such as CaLu3, RT4, HEK293, A549 and U-251MG. An important relationship was observed between expression levels, protein diffusivity, and thermodynamically favorable interactions between host proteins and the viral spike, suggesting potential sites of early infection other than the lungs. This research is expected to stimulate future quantitative experiments and promote systematic investigation of the effect of crowding presented here.

Gasometría arterial en residentes a gran altura, El Alto ­ Bolivia 2020 / Arterial blood gas analysis in residents at high altitude, El Alto, Bolivia 2020

INTRODUCCIÓN: la presión barométrica determina la presión parcial de los gases tanto en el medio ambiente como a nivel alveolar pulmonar, por lo que, para una determinada presión barométrica, la presión de oxígeno y dióxido de carbono es distinta. OBJETIVO: el objetivo del presente estudio es caracterizar los valores de gasometría arterial en residentes adultos sanos a muy alta altitud en la ciudad de El Alto. METODOLOGÍA: Estudio serie de casos, realizado durante la gestión 2019 en 22 (73%) mujeres y 8 varones (27%), con una media de edad de 36.07 años. El estudio contó con la aprobación del Comité de Ética Hospitalario. RESULTADOS: a 4150 metros sobre el nivel del mar, el pH tiende hacia la alcalosis (7.43) así como la PaO2 es menor (58.69mmHg), el valor de PaCO2 (26.14mmHg) es menor correspondiente a una ciudad con mayor altitud, el valor de bicarbonato sérico (20.14mmol/L) se encuentra disminuido en compensación a la disminución de la PaCO2 y el valor de la SatO2% (91.7%) es menor al reportado a nivel del mar. Hay que destacar que el índice PaO2/FiO2 no concuerda con el cálculo realizado según la fórmula propuesta dentro de la definición de los Criterios de Berlín. Se evidencia también que, en la muestra, si bien la correlación entre la PaO2 y la SatO2% resulta como alta, no resulta perfectamente lineal. CONCLUSIÓN: resulta imperativo precisar las características propias a la fisiología correspondientes a cada altitud, buscando aplicar parámetros propios como los de la gasometría arterial para el tratamiento de las enfermedades prevalentes en cada región, así como desarrollar investigaciones específicas a gran altura, ensayar posibilidades y documentarlas.

INTRODUCTION: barometric pressure determines the partial pressure of gases both in the environment and at the pulmonary alveolar level, so for a given barometric pressure, the pressure of oxygen and carbon dioxide is different. The objective of this paper is to characterize arterial blood gas values in healthy adult residents at very high altitude in the city of El Alto. METHODOLOGY: case series study, carried out during 2019; 22 volunteers (73%) women and 8 men (27%), mean age 36.07 years are included. The study was approved by the Hospital Bioethics Committee. RESULTS: at 4150 meter above sea level, pH tends towards alkalosis (7.43) as well as PaO2 is lower (58.69mmHg), the value of PaCO2 (26.14mmHg) is lower corresponding to a city with higher altitude, the value of serum bicarbonate (20.14mmol/L) is lower in compensation at the decrease in PaCO2 and the value of SatO2% (91.7%) is lower than that reported at sea level. The PaO2/FiO2 index does not match the calculation made according to the proposed formula within the definition of the Berlin Criteria. It is also evident that, although the correlation between PaO2 and SatO2% is high, it is not perfectly linear. CONCLUSION: it is imperative to precise the characteristics corresponding to the physiology corresponding to each altitude, looking forward to apply these parameters, such as those of arterial blood gas, in the treatment of prevalent diseases for each region, and so as develop specific studies at high altitude, also testing and reporting them.

A platform to understand amyotrophic lateral sclerosis (ALS) and extend human motor neurons longevity / Plataforma para empezar a comprender la esclerosis lateral amiotrófica (ELA) y extender la longevidad de neuronas motoras humanas

In vitro modeling of neurodegenerative diseases is now possible by using patient-derived induced pluripotent stem cells (iPS). Through them, it is nowadays conceivable to obtain human neurons and glia, and study diseases cellular and molecular mechanisms, an attribute that was previously unavailable to any human condition. Amyotrophic lateral sclerosis (ALS) is one of the diseases that has gained a rapid advance with iPS technology. By differentiating motor neurons from iPS cells of ALS- patients, we are studying the mechanisms underlying ALS- disease onset and progression. Here, we introduce a cellular platform to help maintain longevity of ALS iPS-motor neurons, a cellular feature relevant for most late-onset human diseases. Long term cultures of patient-derived iPS cells might prove to be critical for the development of personalized-drugs.

Actualmente es posible modelar in vitro enfermedades neurodegenerativas humanas mediante el uso de células madre pluripotentes inducidas (iPS) derivadas del paciente. A través de ellas, es hoy concebible obtener neuronas y glía humanas, y estudiar mecanismos celulares y moleculares de enfermedades, un atributo que anteriormente no era posible para ninguna condición humana. La esclerosis lateral amiotrófica (ELA) es una de las enfermedades que se ha beneficiado con la tecnología de iPS. Al diferenciar neuronas motoras de células iPS obtenidas de pacientes con ELA, hemos iniciado estudios sobre los mecanismos que subyacen a la aparición y progresión de la enfermedad. Aquí, presentamos el desarrollo de una plataforma celular que permite extender la longevidad de las neuronas motoras derivadas de iPS, una característica relevante para la mayoría de las enfermedades humanas de inicio tardío. Los cultivos a largo plazo de células iPS provenientes de pacientes pueden ser determinantes en el desarrollo de terapias asociadas a la medicina de precisión.

Differences between diabetic and non-diabetic patients with community-acquired pneumonia in primary care in Spain.

BACKGROUND: Diabetes is one of the underlying risk factors for developing community-acquired pneumonia (CAP). The high prevalence of diabetes among population and the rising incidence of this illness, converts it as an important disease to better control and manage, to prevent its secondary consequences as CAP. The objective of this research is to describe the characteristics of the patients with diabetes and the differences with the no diabetes who have had an episode of CAP in the context of the primary care field. METHODS: A retrospective, observational study in adult patients (> 18 years-old) who suffer from CAP and attended at primary care in Spain between 2009 and 2013 was developed using the Computerized Database for Pharmacoepidemiological Studies in Primary Care (BIFAP). We carried out a descriptive analysis of the first episodes of CAP, in patients with or without diabetes as comorbidity. Other morbidity (CVA, Anaemia, Arthritis, Asthma, Heart disease, Dementia, Depression, Dysphagia, Multiple sclerosis, Epilepsy, COPD, Liver disease, Arthrosis, Parkinson's disease, Kidney disease, HIV) and life-style factors were also included in the study. RESULTS: A total of 51,185 patients were included in the study as they suffer from the first episode of CAP. Of these, 8012 had diabetes as comorbidity. There were differences between sex and age in patients with diabetes. Patients without diabetes were younger, and had less comorbidities including those related to lifestyles such as smoking, alcoholism, social and dental problems than patients with diabetes. CONCLUSIONS: Patients who developed an episode of CAP with diabetes have more risk factors which could be reduced with an appropriate intervention, including vaccination to prevent successive CAP episodes and hospitalization. The burden of associated factors in these patients can produce an accumulation of risk. Health care professional should know this for treating and control these patients in order to avoid complications. Diabetes and those other risk factors associated could be reduced with an appropriate intervention, including vaccination to prevent the first and successive CAP episodes and the subsequent hospitalization in severe cases.

Experimental determination of the gadolinium dose enhancement in phantom irradiated with low energy X-ray sources by a spectrophotometer -Gafchromic-EBT3 dosimetry system.

This work reports the experimental determination of dose enhancement produced in phantoms containing target volumes doped with Gadolinium and irradiated with low-energy X-ray beams. EBT3 Gafchromic films were immersed into 5 ml target volumes to simulate tumor filling with 100% ultra-pure water (blank sample) and ultra-pure water infused with Gd solution (Omniscan®) in different concentration (9-24 mg/ml). The dose enhancement due to excitation of Gd K-edge (50.2 keV) was evaluated in terms of the increment in optical density, obtained by a Spectrophotometer-Gafchromic-EBT3 dosimetry system calibrated in terms of the dose-response for 1-8 Gy range. The dose enhancement was evaluated in two condition: a beam quality with spectrum above the absorption edge in a medium with and without Gd; and two beam qualities (above and below the absorption edge) in a medium with a fixed Gd concentration (18 mg/ml). The obtained results confirmed increments in relative dose enhancement according to Gd concentrations up to 18 mg/ml, with a dose enhancement of 1.1 Gy and an average percentage enhancement of 28.4%. For higher values of concentration, the attenuation interaction of the primary beam is more relevant instead of dose enhancement process. On the other hand, the dose enhancement obtained to comparison the spectra above and below the absorption edge, shown results up to 3.3 Gy of enhancement and average percentage enhancement of 88%.

Insuficiencia ovárica prematura / Premature ovarian insufficiency

RESUMEN La insuficiencia ovárica prematura es la pérdida de la función ovárica antes de los 40 años de edad. Se caracteriza por hipogonadismo hipergonadotrófico y amenorrea u oligomenorrea. Su etiología es multifactorial, pudiendo deberse a causas iatrogénicas, genéticas, metabólicas, autoinmunes y ambientales; siendo de origen idiopático en el 90 % de los casos. Su incidencia es de 1 cada 100 mujeres menores de 40 años y 1 cada 1000 mujeres menores de 30 años. En la actualidad no existe un único marcador que se pueda utilizar para calcular la reserva ovárica; sin embargo, en los últimos años la hormona antimülleriana ha demostrado presentar algunas ventajas respecto a los biomarcadores clásicamente utilizados. Además, diversos estudios indican que existe una correlación positiva entre los niveles de esta hormona y el recuento de folículos antrales, que es, por el momento, el método más confiable para evaluar reserva ovárica debido a las actuales dificultades técnicas para la determinación de hormona antimülleriana.

ABSTRACT Premature ovarian insufficiency, the loss of ovarian function before the age of 40 years, is characterized by hipergonadotrofic hipogonadism and amenorrhea or oligomenorrhea. The etiology is multifactorial, and can be due to genetic, metabolic, autoimmune, environmental or iatrogenic causes, being idiopathic 90% of cases. Currently there is not a single marker that can be used for estimate ovarian reserve in this patients; however, in recent years antimüllerian hormone has proved to have some advantages over other classical biomarkers. Moreover, several studies indicate a positive correlation between antimüllerian hormone concentration and antral follicle count, considered nowadays the most reliable method for ovarian reserve estimation.

Efecto de un dispositivo binivel autoajustable sobre los eventos respiratorios en pacientes con Síndrome de Apnea Obstructiva del Sueño / Effect of an auto-bilevel device on respiratory events in patients with obstructive sleep Apnea syndrome

RESUMEN Objetivo: Evaluar el efecto de un dispositivo binivel autoajustable sobre los eventos respiratorios en pacientes adultos con síndrome de apnea obstructiva del sueño (SAOS) grave. Se revisaron las polisomnografías (PSG) realizadas con el uso de un dispositivo de binivel autoajustable. La arquitectura de sueño, eventos respiratorios, saturación de O2 (SpO2) y dióxido de carbono exhalado (EtCO2) se compararon entre la PSG basal y la PSG terapéutica. Resultados: Se incluyeron 10 PSG. El dispositivo binivel autoajustable corrigió la arquitectura de sueño; disminuyó el índice de apnea hipopnea (IAH) de 76 (39-137) a 14 (6-13) a expensas de apneas obstructivas y mixtas (p < 0,05), no se observó descenso significativo en las hipopneas. Las apneas centrales incrementaron de 0,5 (0-12,4) a 8,2 (0-20) h−1. La SpO2 y EtCO2 mejoraron. Conclusiones: En pacientes con SAOS grave el dispositivo binivel autoajustable corrige la arquitectura de sueño, mejora la SpO2 y EtCO2 y disminuye el IAH a expensas de apneas obstructivas y mixtas, pero podría no eliminar las hipopneas e incrementar las apneas centrales.

Objective: To evaluate the effect of an auto-bilevel device on respiratory events in adults with severe obstructive sleep apnea syndrome (OSAS). Polysomnographies (PSG) with the use of auto-bilevel device were reviewed. Sleep architecture, respiratory events, O2 saturation (SpO2) and exhaled carbon dioxide (EtCO2) were compared among baseline and therapeutic PSG. Results: We included 10 PSG. Auto-bilevel device corrected the sleep architecture; the apnea hypopnea index (AHI) decreased from 76 (39-137) to 14 (6-13) at the expense of obstructive and mixed apneas (p < 0.05), there was no significant decrease in hypopneas. Central apneas increased from 0.5 (0-12.4) to 8.2 (0-20) h−1. SpO2 and EtCO2 improved. Conclusions: In severe OSAS auto-bilevel device corrects sleep architecture, improves SpO2 and EtCO2 and decreases AHI at the expense of obstructive and mixed apneas, but could not eliminate hypopneas and even could increase central apneas.

Evaluation of an Online Educational Program for Parents and Caregivers of Children With Food Allergies.

BACKGROUND AND OBJECTIVE: The increasing prevalence of food allergy affects both patients and their families. Objective: The aim of this study was to evaluate the impact of an online educational program designed for parents and caregivers of children with food allergies. The program was developed by a multidisciplinary group comprising health care professionals, researchers, and expert patients under the participatory medicine model. MATERIAL AND METHODS: Participants took a 2-week online educational program covering major topics in food allergy management. General knowledge about the disease, symptoms, treatment, and topics relevant to families' daily lives were evaluated. The contents included educational videos, online forums, and live video chats. A pretest/posttest questionnaire survey was used to evaluate the impact of the program. RESULTS: A total of 207 participants enrolled in the educational program, which was completed by 130 (62.8%). Knowledge acquisition improved significantly following participation in the program in 15 out of 30 items (50%), reaching P<.001 for 8 items (26.7%). Of the 207 participants who started the program, 139 (67.1%) visited online forums, and 27.5% attended video chats. Average overall satisfaction with the educational program was 8.78 (on a scale of 0 to 10). CONCLUSIONS: The results obtained show that parents improved their knowledge in all areas of food allergy. The high level of satisfaction among participants suggests that digital learning tools are effective and motivational, enabling patients to acquire appropriate knowledge and thus increasing their quality of life.

Lower tier toxicity risk assessment of agriculture pesticides detected on the Río Madre de Dios watershed, Costa Rica.

Costa Rica is a tropical country with one of the highest biodiversity on Earth. It also has an intensive agriculture, and pesticide runoff from banana and pineapple plantations may cause a high toxicity risk to non-target species in rivers downstream the plantations. We performed a first tier risk assessment of the maximum measured concentrations of 32 pesticides detected over 4 years in the River Madre de Dios (RMD) and its coastal lagoon on the Caribbean coast of Costa Rica. Species sensitivity distributions (SSDs) were plotted in order to derive HC5 values for each pesticide, i.e., hazard concentrations for 5 % of the species, often used as environmental criteria values in other countries. We also carried out toxicity tests for selected pesticides with native Costa Rican species in order to calculate risk coefficients according to national guidelines in Costa Rica. The concentrations of herbicides diuron and ametryn and insecticides carbofuran, diazinon, and ethoprophos exceeded either the HC5 value or the lower limit of its 90 % confidence interval suggesting toxic risks above accepted levels. Risk coefficients of diuron and carbofuran derived using local guidelines indicate toxicity risks as well. The assessed fungicides did not present acute toxic risks according to our analysis. Overall, these results show a possible toxicity of detected pesticides to aquatic organisms and provide a comparison of Costa Rican national guidelines with more refined methods for risk assessment based on SSDs. Further higher tier risk assessments of pesticides in this watershed are also necessary in order to consider pesticide water concentrations over time, toxicity from pesticide mixtures, and eventual effects on ecosystem functions.

Purity of islet preparations and 5-year metabolic outcome of allogenic islet transplantation.

In allogenic islet transplantation (IT), high purity of islet preparations and low contamination by nonislet cells are generally favored. The aim of the present study was to analyze the relation between the purity of transplanted preparations and graft function during 5 years post-IT. Twenty-four patients with type 1 diabetes, followed for 5 years after IT, were enrolled. Metabolic parameters and daily insulin requirements were compared between patients who received islet preparations with a mean purity <50% (LOW purity) or ≥50% (HIGH purity). We also analyzed blood levels of carbohydrate antigen 19-9 (CA 19-9)-a biomarker of pancreatic ductal cells-and glucagon, before and after IT. At 5 years, mean hemoglobin A1c (HbA1c levels) (P = .01) and daily insulin requirements (P = .03) were lower in the LOW purity group. Insulin independence was more frequent in the LOW purity group (P < .05). CA19-9 and glucagon levels increased post-IT (P < .0001) and were inversely correlated with the degree of purity. Overall, our results suggest that nonislet cells have a beneficial effect on long-term islet graft function, possibly through ductal-to-endocrine cell differentiation. ClinicalTrial.gov NCT00446264 and NCT01123187.

Salivary epidermal growth factor correlates with hospitalization length in rotavirus infection.

BACKGROUND: The IFI27 interferon gene expression has been found to be largely increased in rotavirus (RV)-infected patients. IFI27 gene encodes for a protein of unknown function, very recently linked to epidermal proliferation and related to the epidermal growth factor (EGF) protein. The EGF is a low-molecular-weight polypeptide that is mainly produced by submandibular and parotid glands, and it plays an important physiological role in the maintenance of oro-esophageal and gastric tissue integrity. Our aim was to determine salivary EGF levels in RV-infected patients in order to establish its potential relationship with IFI27 increased expression and EGF-mediated mucosal protection in RV infection. METHODS: We conducted a prospective comparative study using saliva samples from 27 infants infected with RV (sampled at recruitment during hospital admission and at convalescence, i.e. at least 3 months after recovery) and from 36 healthy control children. RESULTS: Median (SD) EGF salivary concentration was 777 (529) pg/ml in RV-infected group at acute phase and 356 (242) pg/m at convalescence, while it was 337 (119) pg/ml in the healthy control group. A significant association was found between EGF levels and hospitalization length of stay (P-value = 0.022; r2 = -0.63). CONCLUSIONS: The salivary levels of EGF are significantly increased during the acute phase of natural RV infection, and relate to length of hospitalization. Further assessment of this non-invasive biomarker in RV disease is warranted.

Discovering key residues of dengue virus NS2b-NS3-protease: New binding sites for antiviral inhibitors design.

The NS2B-NS3 protease is essential for the Dengue Virus (DENV) replication process. This complex constitutes a target for efficient antiviral discovery because a drug could inhibit the viral polyprotein processing. Furthermore, since the protease is highly conserved between the four Dengue virus serotypes, it is probable that a drug would be equally effective against all of them. In this article, a strategy is reported that allowed us to identify influential residues on the function of the Dengue NS2b-NS3 Protease. Moreover, this is a strategy that could be applied to virtually any protein for the search of alternative influential residues, and for non-competitive inhibitor development. First, we incorporated several features derived from computational alanine scanning mutagenesis, sequence, structure conservation, and other structure-based characteristics. Second, these features were used as variables to obtain a multilayer perceptron model to identify defined groups (clusters) of key residues as possible candidate pockets for binding sites of new leads on the DENV protease. The identified residues included: i) amino acids close to the beta sheet-loop-beta sheet known to be important in its closed conformation for NS2b ii) residues close to the active site, iii) several residues evenly spread on the NS2b-NS3 contact surface, and iv) some inner residues most likely related to the overall stability of the protease. In addition, we found concordance on our list of residues with previously identified amino acids part of a highly conserved peptide studied for vaccine development.

Inducing ovulation with oestradiol cypionate allows flexibility in the timing of insemination and removes the need for gonadotrophin-releasing hormone in timed AI protocols for dairy cows.

The effects of addition of gonadotrophin-releasing hormone (GnRH) to a progesterone plus oestradiol-based protocol and timing of insemination in Holstein cows treated for timed AI (TAI) were evaluated. Cows (n=481) received a progesterone device and 2mg oestradiol benzoate. After 8 days, the device was removed and 25mg dinoprost was administered. Cows were allocated to one of three (Study 1; n=57) or four (Study 2; n=424) groups, accordingly to ovulation inducer alone (Study 1; oestradiol cypionate (EC), GnRH or both) or ovulation inducer (EC alone or combined with GnRH) and timing of insemination (48 or 54h after device removal; Study 2). In Study 1, the diameter of the ovulatory follicle was greater for GnRH than EC. Oestrus and ovulation rates were similar regardless of ovulatory stimuli. However, time to ovulation was delayed when GnRH only was used. In Study 2, cows treated with GnRH or not had similar pregnancy per AI (P/AI) 30 days (41.5% vs 37.3%; P=0.28) and 60 days (35.9% vs 33.0%; P=0.61) after TAI. TAI 48 and 54h after device removal resulted similar P/AI at 30 days (40.3% vs 38.5%; P=0.63) and 60 days (33.8% vs 35.1%; P=0.72). Thus, adding GnRH at TAI does not improve pregnancy rates in dairy cows receiving EC. The flexibility of time to insemination enables TAI of a large number of cows using the same protocol and splitting the time of AI.

Insertion of the LINE-1 element in the C-MYC gene and immunoreactivity of C-MYC, p53, p21 and p27 proteins in different morphological patterns of the canine TVT / Inserção do elemento LINE - 1 no gene C-MYC e imunorreatividade das proteinas C-MYC, p53, p21 e p27 nos diferentes padrões morfológicos do tumor TVT

The canine transmissible venereal tumor (TVT) affects the external genitalia of dogs by the natural transplant of viable tumor cells. Thus, this research aimed to diagnose and characterize TVT morphological patterns, identify the insertion of the LINE-1 element in C-MYC gene, by means of the polymerase chain reaction (PCR), and evaluate the immunohistochemical expression of C-MYC, p53, p21 and p27 proteins. The relationship between C-MYC and p53 proteins and their interference on the expression of p21 and p27 were also studied. For that, 20 samples of naturally occurring TVT were used, subjected to cytopathological, histopathological and immunohistochemical analysis, and to molecular diagnosis of neoplasia. The increased tissue expression and the correlation among C-MYC, p53, p21 and p27 proteins indicate reduction and/or loss of their functionality in the TVT microenvironment, with consequent apoptotic suppression, maintenance of cell growth and progression of neoplasia.(AU)

O tumor venéreo transmissível canino (TVT) afeta a genitália externa de cães pelo transplante natural de células tumorais viáveis. Assim, esta pesquisa teve como objetivo diagnosticar e caracterizar TVT em padrões morfológicos, identificar a inserção do elemento LINE-1 em gene C-MYC, por meio da reação em cadeia da polimerase (PCR), e avaliar a expressão imuno-histoquímica do C-MYC, p53, p21 e p27. A relação entre C-MYC e as proteínas p53 e a sua interferência na expressão de p21 e p27 foram também estudadas. Para isso, foram utilizadas 20 amostras de ocorrência natural de TVT, submetido a exame citopatológico, histopatológica e imuno-histoquímica e ao diagnóstico molecular de neoplasia. A expressão aumentada do tecido e a correlação entre a C-MYC e as proteínas p53, p21 e p27 indicam redução e/ou perda de funcionalidade na TVT em seu microambiente, com consequente supressão apoptótica, manutenção do crescimento celular e progressão da neoplasia.(AU)

Evaluation of an educational programme with workshops for families of children with food allergies.

BACKGROUND: When a child is diagnosed with a food allergy, prevention and patient education are the key interventions for maintaining the child's health and quality of life and that of his or her whole family. However, health education activities for the families of children with food allergies are very limited, and most of these activities have not been evaluated. Therefore, the objectives of the present study were to develop an educational programme, to empower its participants through workshops, and to evaluate its results. METHODS: Several types of educational materials were created specifically for the programme, called "Proyecto CESA" ("STOP-FAR Project: Stop Food-Induced Allergic Reactions"). The programme was carried out in seven Spanish cities and was attended by parents and caregivers. The workshops were led by physicians specialising in allergies and by expert patients. Afterwards, participant learning and satisfaction were evaluated based on questionnaires that were completed both before and after the workshops. RESULTS: A significant improvement was observed in 29 items out of 40 (McNemar's test). Participant satisfaction with the programme was also very high: 90% rated the course between 8 and 10 points out of a possible 10 (41% rated it as a 10). CONCLUSION: The face-to-face workshops, which included utilisation of educational materials, had positive results in terms of learning as well as in levels of satisfaction in participating families.

Left Ventricular Dysfunction After Lung Transplantation for Pulmonary Arterial Hypertension.

BACKGROUND: Lung transplantation (LT) is the final treatment option for patients with pulmonary arterial hypertension (PAH). Perioperative challenges after LT are unique and commonly include excessive bleeding, arrhythmias, and primary graft dysfunction. Transient left ventricular dysfunction (LVD) is a known postoperative complication, but not fully explored. We describe our experiences at a single institution. METHODS: We reviewed our database for patients with PAH who underwent LT from July 2008 to July 2012. The data were analyzed for preoperative inotrope use, intravenous prostacyclin, cardiac catheterization, and imaging. Also measured were perioperative ischemic time, bypass time, primary graft dysfunction, ventilator days, length of stay, and mortality. LVD is defined as acute cardiopulmonary compromise (acute worsening of hypoxia with new bilateral infiltrates on imaging) with a drop in LV systolic function of 15% from baseline. We compared data between patients with LVD and without LVD. RESULTS: Sixteen patients met the criteria, the majority of patients (10) with World Health Organization (WHO) group 1 PAH. Thirteen received intravenous prostacyclin therapy, and 6 required inotropes before surgery. Five patients (31%) developed LVD after transplantation. Average time to onset of LVD was 4.2 days. Preoperative vasopressors were required in 60% of those developing LVD. Patients with LVD had lower right and left ventricular ejection fraction with higher left ventricular end diastolic volume before surgery. All patients recovered from LVD within 4 months after LT. CONCLUSIONS: LVD is a phenomenon observed mostly in patients with WHO group 1 PAH receiving LT. Prompt recognition and treatment of this condition reduced morbidity.

Development of an IP-Free Biotechnology Platform for Constitutive Production of HPV16 L1 Capsid Protein Using the Pichia pastoris PGK1 Promoter.

The human papillomavirus (HPV) L1 major capsid protein, which forms the basis of the currently available vaccines against cervical cancer, self-assembles into virus-like particles (VLPs) when expressed heterologously. We report the development of a biotechnology platform for HPV16 L1 protein expression based on the constitutive PGK1 promoter (PPGK1) from the methylotrophic yeast Pichia pastoris. The L1 gene was cloned under regulation of PPGK1 into pPGKΔ3 expression vector to achieve intracellular expression. In parallel, secretion of the L1 protein was obtained through the use of an alternative vector called pPGKΔ3α, in which a codon optimized α-factor signal sequence was inserted. We devised a work-flow based on the detection of the L1 protein by dot blot, colony blot, and western blot to classify the positive clones. Finally, intracellular HPV VLPs assembly was demonstrated for the first time in yeast cells. This study opens up perspectives for the establishment of an innovative platform for the production of HPV VLPs or other viral antigens for vaccination purposes, based on constitutive expression in P. pastoris.

Carcinoma neuroendocrino de vejiga en mujer con infección por VIH: presentación del primer caso mundial / Neuroendocrine carcinome of the bladder in a female with HIV infection: presentation of the first case worldwide

El carcinoma neuroendocrino primario de vejiga es una neoplasia infrecuente que representa el 0,5por ciento de todos los tumores vesicales. La asociación de carcinoma neuroendocrino de vejiga en un paciente con infección por VIH nunca hasta hoy había sido descrita. Presentamos el primer caso clínico español y mundial de esta desconocida y nunca descrita asociación. MATERIAL Y MÉTODOS: Se presenta el caso clínico de una paciente de 46 años con infección por VIH que desarrolló un carcinoma neuroendocrino de vejiga urinaria de evolución fatal. Se describe su clínica de presentación, métodos de diagnóstico utilizados y su tratamiento. La paciente debutó con retención urinaria aguda que rápidamente progresó a la instauración de una uropatía obstructiva alta con deterioro de la función renal. El diagnóstico se efectuó mediante TAC, resección transuretral y estudio histopatológico donde la clave del diagnóstico fue el estudio inmunohistoquímico intensamente positivo para la cromogranina A. El tratamiento adyuvante con quimioterapia le ocasionó una aplasia medular severa, falleciendo por fallo multiórganico a los 26 días de su diagnóstico. A propósito de este caso, se revisa la literatura inglesa en PubMed sobre carcinoma neuroendocrino de vejiga y sobre tumores vesicales en pacientes con infección VIH, no existiendo ningún caso publicado de carcinoma neuroendocrino de vejiga en un paciente con infección por VIH. CONCLUSIONES: El carcinoma neuroendocrino de vejiga es un tumor infrecuente y muy agresivo. Es un tumor que suele presentarse clínicamente en estadios avanzados o metastásicos donde ninguna terapia es eficaz. El tratamiento incluye resección trans-uretral (RTU), cistectomía parcial, cistectomía radical y quimioterapia. El estudio inmunohistoquímico (IHQ) y la tinción con cromogranina A dan la clave para su diagnóstico. Su presentación en pacientes VIH implica muy mal pronóstico. Éste caso es el primer caso mundial publicado de carcinoma neuroendocrino...

The primary neuroendocrine carcinome of the bladder is an infrequent neoplasm which represents 0.5 percent of all vesical tumors. The association of neuroendocrine carcinome of the bladder in a patient with HIV infection has never been described before today. We present the first clinical case in the Spanish-speaking world and worldwide, of this unknown and never written about association. MATERIAL AND METHODS: The clinical case of a 46-yearoldpatient with HIV infection who developed a neuroendocrine carcinoma of the urinary bladder with a fatal evolution, its clinical presentation, the diagnosis methods used and its treatment, are described. The patient started with a severe urinary retention which rapidly progressed to the establishment of a high obstructive uropathy with deterioration in the renal function. The diagnosis was done using TAC, transurethral resection and histopathological study where the key to diagnosis was the intensely positive immunohistochemical study for the chromogranin A. The adjuvant treatment with chemotherapy led to a severe medular aplasia, with the patient dying due to a multi-organ failure, 26 days after her diagnosis. As a result of this case, English literature on the matter in PubMed about neuroendocrine carcinome of the bladder and about vesical tumors in patients with HIV infection was revised, with no published case existing about neuroendocrine carcinome in a patient with HIV. CONCLUSIONS: The neuroendocrine carcinome of the bladder is an infrequent and very aggressive tumor. It is a tumor that tends to be clinically present in advanced or metastasic states, where no therapy is efficient. The treatment includes transurethral resection (TUR), partial cystectomy, radical cystectomy and chemotherapy. The immunohistochemical study (IHC), and the stain with chromogranin A are key for its diagnosis. Its presentation in HIV patients implies a very bad prognosis. This case is the first published case worldwide of neuroendocrine...

Theoretical indications on the relationship between pyrogallol[4]arenes dynamics of assembling and geometry / Indicios teóricos entre la relación dinámica de ensamblaje y geometría de pirogalol[4]arenos / Indícios teóricos sobre a relação entre dinâmica de montagem e geometria de pirogalol[4]arenos

Los pirogalol[4]arenos son macrociclos con gran potencial como bloques de construcción de nanocápsulas. Los dímeros precursores del 2,8,14,20-tetrametilpirogalol[4]areno y del 2,8,14,20-tetrafenilpirogalol[4]areno se estudiaron teóricamente para obtener entendimiento acerca de la dinámica de ensamble de estos compuestos. Las curvas de energía potencial a lo largo del ángulo de torsión del enlace R-pirogalol)CH-(R-pirogalol) de los dímeros han sido calculadas al nivel de teoría B3LYP/6-311G(d,p). Se encontró que las barreras de energía para rotación libre en torno al enlace seleccionado son 0.00133 Hartrees para el dímero con sustituyentes alquílicos y 0.77879 Hartrees para aquél con sustituyentes arílicos. Estos valores implican que la rotación libre en torno al enlace seleccionado es permitida para el primer dímero pero es prohibida para el segundo. Puesto que las orientaciones del sustituyente y el anillo de pirogalol en torno a este enlace posiblemente determinan la geometría de la estructura final, parece razonable proponer que el pirogalol[4]areno con sustituyentes alquílicos más probablemente adoptará una geometría de corona, mientras que el pirogalol[4]areno con sustituyentes arílicos mas probablemente adoptará una geometría de silla. Estas previsiones están en acuerdo con evidencia experimental la cual demuestra que la geometría de los pirogalol[4]arenos es dependiente del tipo de sustituyentes presentes en ellos.

O pyrogalol [4]arenes são macrocycles com grande potencial como blocos de construção da nanocapsules. Os dímeros precursores de 2,8,14,20-tetrametilpirogalol[4]arene e 2,8,14,20-tetrafenilpirogalol[4]arene foram estudados teoricamente para obter uma compreensão sobre a dinâmica do monte destes compostos. As curvas de energia potencial ao longo do ângulo de torção da link R-pkogdol)CH-(R-pirogalol) dos dímeros foram calculados ao nível da teoria B3LYP/ 6-311G(d,p). Verificou que as barreiras de energia da rotação livre em torno do link selecionado é 0,00133 Hartrees para ao dímero com substituintes alquil e 0,77879 Hartrees para que com substituintes aril. Esses valores implicam que a livre rotação em torno da link seleccionado é permitido para o primeiro dímero mas é proibida para o segundo. Uma vez que a orientação dos substituintes e o anel de pyrogalol em torno deste link possivelmente determinar a geometria da estrutura final, parece razoável sugerir que o pyrogalol[4]arene com substituintes alquil mais provavelmente vai ter uma geometria da copa, enquanto o pyrogalol[4]arene com substituintes resíduo mais provavelmente irá ter uma geometría de cadeira. Estas previsões estão de acordo com evidências experimentais que demonstram que a geometria do pyrogalol[4]arenes é dependente do tipo de substituintes presentes nos mesmos.

Pyrogallol[4]arenes are macrocycles with high potential as building blocks for nanocapsules. We theoretically studied the dimeric precursors of 2,8,14,20-tetramethylpyrogaUol[4]arene and 2,8,10,14-tetraphenylpyrogallol[4] arene to understand the dynamics of assembly of these compounds, and calculated the potential energy curves along the torsion angle of the (R-pyrogallol)CH-(R-pyrogallol) dimeric bond at the B3LYP/6-311G(d,p) level of theory. We found that the energy barriers for free rotation around the selected bond are 0.00133 Hartrees for the alkyl-substituted dimer and 0.77879 Hartrees for the aryl-substituted dimer. These values imply that the free rotation around the selected bond exists for the first dimer but not for the second one. Because the orientation of the substituent and the pyrogallol ring around this bond are likely to determine the geometry of the final structure, we propose that the alkyl-substituted compound will most likely adopt a crown-shaped geometry whereas the aryl-substituted compound will adopt a chair-shaped geometry. These predictions concur with experimental evidence, which shows that the geometry of pyrogallol[4]arenes depends on the substituents attached to them.