Evaluación del perfil de seguridad del medicamento tocilizumab en pacientes sospechosos o confirmados por COVID-19 / Evaluation of the safety profile of the drug tocilizumab in patients suspected or confirmed by COVID-19

Introducción: La enfermedad de COVID-19, es una enfermedad emergente cuya patogénesis se relaciona con la tormenta de citocina, la interleucina 6 juega un papel importante en la tormenta de citocinas. El medicamento tocilizumab, es un anticuerpo monoclonal humanizado, el cual se une al receptor soluble IL-6. En pacientes con COVID-19 se ha observado que el uso de tocilizumab disminuye la inflamación exacerbada. Ante este nuevo uso del medicamento es relevante establecer el balance beneficio-riesgo en estos pacientes con COVID-19, identificando con ello las reacciones adversas a medicamentos que pueden estar relacionadas al uso de tocilizumab. Materiales y métodos: Estudio de farmacovigilancia descriptivo y transversal en una cohorte retrospectiva en pacientes sospechosos o confirmados por COVID-19 en el Instituto Nacional de Cardiología Ignacio Chávez de la Ciudad de México, México en el periodo 05 de mayo del 2020 al 20 de enero del 2021. Resultados: De los 36 pacientes participantes en este estudio, la edad promedio fue 53 años, de los cuales 30 fueron hombres y 6 fueron mujeres. Las comorbilidades identificadas en este estudio fue la hipertensión arterial sistémica, seguida de la diabetes mellitus tipo II. En la evaluación de los estudios de laboratorio se observó que 2 pacientes desarollaron neutropenia moderada, mientras que en 5 pacientes se identificó trombocitopenia leve y 2 pacientes desarrollaron trombocitopenia moderada. Las infecciones bacterianas identificadas en el estudio con el uso del medicamento fueron: 5 aislamientos de Klebsiella oxytoca, 4 a Escherichia coli y 4 a Pseudomonas aeruginosa... (AU)

Introduction: COVID-19 is a new emerging disease which pathogenesis is mediated by a cytokines storm, interleukin 6 plays an important part of this storm. Tocilizumab is a humanized monoclonal antibody that binds to the IL-6 receptor. In patient with COVID-19, exacerbated inflammation has been observed to decrease when given tocilizumab. Due to the new use of this drug is relevant to stablish the risk-benefit ratio in COVID-19 patients, by identifying the drug adverse reactions that may be related to the use of tocilizumab. Material and methods: Descriptive and cross-sectional pharmacovigilance study in a retrospective cohort in patients suspected or confirmed by COVID-19 in the National Institute of Cardiology in Mexico City, Mexico from May 5, 2020 to January 20, 2021. Outcomes: From 36 patients in this study, the average age was 53 years of which 30 were men and 6 were women. The comorbidities identified in this study were systemic arterial hypertension followed by type II diabetes mellitus. Evaluating the laboratory results we observed 2 patients developed moderate neutropenia, 5 patients presented mild thrombocytopenia and 2 patients moderate thrombocytopenia. The bacterial infections identified in the study with the use of the tocilizumab were: 5 isolates Klebsiella oxytoca, 4 isolates Escherichia coli and 4 Pseudomonas aeruginosa. Conclusion: Knowing the possible drug adverse reactions that occurred in patients with COVID-19 who were administered tocilizumab, allow us to the identify the risks associated with the drug, determining the safety profile and be alert of bacterial infections, neutropenia, and thrombocytopenia, throughout a pharmacotherapeutical follow up, thereby identifying possible associated alterations possibly restated with the use of tocilizumab. (AU)

Cirugía periapical y regeneración: revisión sistemática / Periapical surgery and regeneration: systematic review

Introducción: Tras un cirugía periapical, suele estar asociado un defecto óseo que puede ser regenerado o no según la literatura. El objetivo es analizar si la regeneración ósea asociada a la cirugía periapical tiene efectos beneficiosos en la curación de los diferentes defectos. Material y Mètodos: Revisión sistemática realizada en Medline-Pubmed, Scopus y Cochrane. Se consideraron criterios PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses). Ensayos clínicos aleatorizados, estudios de casos y controles, de cohortes y transversales publicados entre enero 2011 y abril 2022, que analizaran el èxito de la cirugía periapical combinada o no con diferentes tècnicas de regeneración ósea, idioma inglès o español, realizados en humanos y adultos (≥16años) fueron incluidos. Resultados: Se incluyeron un total de 8 artículos que analizaban una "n" total de 285 sujetos con 35 abandonos. Se analizan diferentes tipos de lesiones: lesiones transversales, defectos apicomarginales y lesiones periapicales, excepto un estudio que compara las tres. Se utilizan diferentes materiales de regeneración. Se obtuvo un rango de èxito de regeneración ósea entre 81,6% y 93,7%. Discusión: Los avances en equipos de magnificación visual aumento, instrumentos y materiales endodónticos son la razón de un mayor èxito de las cirugías periapicales. Las imágenes en 3D, son valiosas para obtener un mejor diagnóstico del tamaño y tipo de defecto, y planificar así el tratamiento considerando la necesidad de regenerar o no. Aunque los resultados siguen siendo inciertos parece ser que en los defectos transversales y apicomarginales es mejor regenerar, al contrario de los defectos periapicales. (AU)

Introduction: According to the literature, after a periapical surgery there usually remains a bone defect that may be regenerated or not. The aim of this review is to analyse whether bone regeneration associated with periapical surgery is favourable on the healing of different defects. Material and Methods: A systematic review was performed in Medline-Pubmed, Scopus and Cochrane. PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) criteria were considered. Randomized clinical trials, case-control, cohort and cross-sectional studies published between January 2011 and April 2022, that analyses the success of periapical surgery associated or not with different bone regeneration techniques, English or Spanish, humans and adults (≥ 16years) studies were included. Results: A total of eightreports were included. They evaluated 285 subjects, with 35 subjects that dropped out. Different types of lesions were analysed in each report: through-and-through lesions, apico-marginal lesions, and periapical lesions, except for one that compared all of them. Different kinds of bone regenerative materials were used. A range of success of between 81,6% and 93,7% of bone regeneration was achieved. Discussion: Improvements in augmentation equipment, instruments, and materials are the reason for the large success of periapical surgery. 3D images are useful to make a better diagnosis of the size and kind of the defect and, in this way, to plan the treatment considering the need to regenerate or not. Although the results remain uncertain, it seems that for through-and-thought and apico-marginal lesions, it is better to regenerate in contrast with periapical lesions. (AU)

Corticosteroid safety in COVID-19 patients: a pharmacovigilance study / Seguridad de corticoesteroides en pacientes con COVID-19: un estudio de farmacovigilancia

The use of oral and intravenous corticosteroids as a treatment for SARS-CoV-2 infection has been shown to inhibit the exaggerated inflammatory response, reducing symptoms and days of hospitalization of patients. However, its use is controversial because not enough clinical studies have been made to verify the safety of the drugs. Objective: To assess the safety profile of corticosteroids treatment, at high and low doses, in suspected or confirmed patients with COVID-19, determining the most frequent side effects in patients, and assessing whether the administration of the drugs represents a greater benefit than the risk of presenting these effects. Methods: Ambispective study of active pharmacovigilance at a cohort of confirmed or suspected COVID-19 patients, treated with intravenous and oral corticosteroids. 366 patients were evaluated and divided into 3 groups: use of methylprednisolone (155 mg average) every 24 hours for 3 days, dexamethasone (6 mg) every 24 hours for 10 days, and a control group. Results: The distribution of the cases with hyperglycemia was 33 in high doses and 82 with low doses of corticosteroids and both high and low doses have a similar distribution in cases of infections. When evaluating the harshness and severity of hyperglycemia in the two groups with corticosteroids, it is observed that patients with high doses present more harsh (48%). In case of harshness and severity of infections it is observed that patients with high doses present more harsh (62%) and more severe (79%) cases than those who were administered low doses. (AU)

El uso de corticoides orales e intravenosos como tratamiento para la infección por SARS-CoV-2 ha demostrado inhibir la respuesta inflamatoria exagerada, reduciendo los síntomas y los días de hospitalización de los pacientes. Sin embargo, su uso es controvertido porque no se han realizado suficientes estudios clínicos para verificar la seguridad de los medicamentos. Objetivo: Evaluar el perfil de seguridad del tratamiento con corticoides, a dosis altas y bajas, en pacientes con sospecha o confirmación de COVID-19, determinando los efectos secundarios más frecuentes en los pacientes, y valorando si la administración de los fármacos representa un mayor beneficio que el riesgo de presentar estos efectos. Métodos: Estudio ambispectivo de farmacovigilancia activa en una cohorte de pacientes confirmados o sospechosos de COVID-19, tratados con corticoides intravenosos y orales. Se evaluaron 366 pacientes y se dividieron en 3 grupos: uso de metilprednisolona (155 mg promedio) cada 24 horas por 3 días, dexametasona (6 mg) cada 24 horas por 10 días y un grupo control. Resultados: La distribución de los casos con hiperglucemia fue de 33 casos usando dosis altas y 82 con dosis bajas de corticoides, tanto las dosis altas como las bajas tienen la misma distribución en los casos de infecciones. Al evaluar la severidad y gravedad de la hiperglucemia en los dos grupos con corticoides, se observa que los pacientes con dosis altas presentan mayor gravedad (48%). En caso de severidad y gravedad de las infecciones se observa que los pacientes con dosis altas presentan casos más graves (62%) y más severos (79%) que los que recibieron dosis bajas. (AU)

Estresores académicos y manifestaciones biopsicosociales en estudiantes universitarios de fisioterapia / Academic stressors and biopsychosocial manifestations in physiotherapy students

Introducción y objetivos: Los estudiantes universitarios se ven constantemente expuestos a situaciones estresantes como parte de las exigencias y demandas de la formación universitaria. El objetivo del presente estudio fue analizar la relación entre factores asociados al estrés académico y variables conductuales, psíquicas y físicas nocivas para el bienestar biopsicosocial en estudiantes universitarios de primer y cuarto año académico de la carrera de fisioterapia. Material y métodos: Se incluyeron 68 estudiantes de ambos géneros (43 de primero y 25 de cuarto año). Las manifestaciones físicas, psíquicas y conductuales se determinaron mediante un cuestionario auto-informado y el estrés académico se midió con el Cuestionario de Estrés Académico-ECEA. Resultados: La sobrecarga académica tuvo una asociación significativa con manifestaciones psíquicas, conductuales y físicas (p<0,005) en estudiantes de primer año, mientras que las deficiencias metodológicas y los exámenes tuvieron un efecto significativo sobre los 3 aspectos biopsicosociales evaluados en los estudiantes de cuarto año (p<0,050). Finalmente, los aspectos vinculados a la metodología docente impactaron negativamente en el bienestar de ambos grupos (p<0,050). Conclusiones: Este estudio aportó información sobre los factores estresores y sus potenciales implicaciones para la salud de los estudiantes, antecedentes que serán de utilidad para abordar el estrés académico de manera sistémica, así como para futuras investigaciones relacionadas con el bienestar biopsicosocial de la comunidad universitaria.

Introduction and objectives: University students are constantly exposed to stressful situations as part of the demands and requirements of university education. The aim of the present study was to analyse the relationship between factors associated with academic stress and behavioural, psychological, and physical variables harmful to biopsychosocial well-being in first and fourth year physiotherapy undergraduates. Material and methods: Sixty-eight students of both genders (43 from the first year and 25 from the fourth year) were included. Physical, psychological, and behavioural manifestations were determined through a self-report questionnaire and academic stress was measured with the Academic Stress Questionnaire-ECEA. Results: Academic overload had a significant association with mental, behavioural and physical manifestations (P<.005) in the first-year students, while methodological deficiencies and exams had a significant effect on the three biopsychosocial aspects evaluated in the fourth-year students (P<.050). Finally, aspects linked to teaching methodology had a negative impact on the well-being of both groups (P<.050). Conclusions: This study provided information on stressors and their potential implications for students’ health, background information that will be useful to address academic stress in a systemic way, as well as for future research related to the biopsychosocial well-being of the university community.

Simuladores en odontología y la formación de habilidades clínicas: un diálogo permanente / Simulators in dentistry and clinical skill training: a permanent dialogue

El avance tecnológico evidencia el gran desarrollo de los simuladores en la educación odontológica. Con el transcurrir de los años, el uso de simuladores como herramienta educativa para el logro de competencias en estudiantes de preclínica ha tenido una creciente incorporación en los planes de estudio. Por ello, el presente artículo tiene como objetivo revisar la historia de los simuladores en la educación odontológica e identificar las habilidades desarrolladas con el uso de simuladores en estudiantes universitarios de pregrado a nivel mundial. Al respecto, se realizó una búsqueda de artículos científicos indexados en bases de datos como Medline, Scopus, LILACS, SciELO, Google académico y Redalyc; se desarrolló la búsqueda bibliográfica hasta enero 2021 y se consideró las siguientes palabras clave: simulación, educación en odontología, estudiantes de odontología, educación basada en competencias. En los resultados se identificó cuatro etapas relevantes como una manera interactiva para explicar la historia de la simulación en odontología, cada una de estas etapas estuvo determinada por el impacto tecnológico de cada época, la necesidad de entrenamiento de los estudiantes y la integración en los planes de estudio. Con esta revisión, se concluyó que existe evidencia que el uso de simuladores durante la formación preclínica universitaria desarrolla habilidades en el estudiante de odontología, y que estas habilidades clínicas están relacionadas con el tipo de simulador usado durante su formación de pregrado.

Technological advance shows the great development of simulators in dental education. The use of simulators as an educational tool to achieve competencies in preclinical students has been increasingly incorporated into the study plans. Therefore, this article aims to review the history of simulators in dental education and identify the skills developed with the use of simulators in undergraduate university students worldwide. In this regard, a search was carried out for scientific articles indexed in databases such as Medline, Scopus, LILACS, SciELO, Academic Google and Redalyc; the bibliographic search was carried out until January 2021 and the following keywords were considered: simulation, dental education, dentistry students, competency-based education. In the results, four relevant stages were identified as an interactive way to explain the history of simulation in dentistry, each of these stages was determined due to the technological impact of each era, the need for training of the students and the integration into the study plans. It was concluded that there is evidence that the use of simulators during preclinical university training develops skills in dental students, these are related to the type of simulator used during their undergraduate training.

Extinction of herbivorous dinosaurs linked to Early Jurassic global warming event.

Sauropods, the giant long-necked dinosaurs, became the dominant group of large herbivores in terrestrial ecosystems after multiple related lineages became extinct towards the end of the Early Jurassic (190-174 Ma). The causes and precise timing of this key faunal change, as well as the origin of eusauropods (true sauropods), have remained ambiguous mainly due to the scarce dinosaurian fossil record of this time. The terrestrial sedimentary successions of the Cañadón Asfalto Basin in central Patagonia (Argentina) document this critical interval of dinosaur evolution. Here, we report a new dinosaur with a nearly complete skull that is the oldest eusauropod known to date and provide high-precision U-Pb geochronology that constrains in time the rise of eusauropods in Patagonia. We show that eusauropod dominance was established after a massive magmatic event impacting southern Gondwana (180-184 Ma) and coincided with severe perturbations to the climate and a drastic decrease in the floral diversity characterized by the rise of conifers with small scaly leaves. Floral and faunal records from other regions suggest these were global changes that impacted the terrestrial ecosystems during the Toarcian warming event and formed part of a second-order mass extinction event.

Evaluation of the biocompatibility of a PVA/SA scaffold with a human gingival fibroblast (HGF) by using electrochemical impedance spectroscopy.

The biocompatibility of human gingival fibroblasts (HGF) was evaluated in different concentrations of poly(vinyl alcohol) and sodium alginate (PVA/SA) nanofibres (3.5 wt% 4 wt% and 5 wt%). The PVA/SA nanofibres were deposited on the surface of an electrode microchip by using the electrospinning technique. Electrochemical impedance spectroscopy (EIS) was applied to measure the dielectric properties of each system. In order to provide a detailed analysis as well as a right physical interpretation of the EIS results, the data was fitted with an electric equivalent circuit based on the EIS and the microscopic assessments. The results registered three different time constants (TCs) of the PVA/SA scaffold which indicated different layers at different depths of the scaffold. The TCs changed their dielectric properties depending on the PVA/SA concentration. The 4 wt% system showed the highest biocompatibility properties, given that its resistance and electrochemical capacitance show the formation of a mature-stage cell interaction of HGF. The EIS data offers an exhaustive analysis of the biological activity of the cell response in real time to determine its biocompatibility features. Fluorescence analysis demonstrated a heterogeneous growth of the HGF on the PVA/SA scaffold surface.

Factores asociados a la malignización del liquen plano oral. Revisión de la literatura / Factors associated with the malignization of oral lichen planus. Review of the literatura

Introducción: El liquen plano (LP) es una enfermedad sistémica autoinmune de etiología desconocida y curso crónico que puede resultar en el desarrollo de carcinoma oral de células escamosas (COCE). Objetivo: Evaluar la evidencia relacionada con la malignización del LPO, así como las características clínicas y la incidencia de las lesiones que pueden desarrollan un COCE. Material y métodos: Se revisa la base de datos MEDLINE PubMed de los últimos 5 años (2013-2018) utilizando las siguientes palabras clave: "malignant transformation of oral lichen planus" and "squamous cell oral carcinoma". Resultados: Se obtuvieron 48 artículos de los cuales tras la lectura resúmenes y la aplicación de nuestros criterios de inclusión y exclusión obtuvimos un total de 19 artículos, además de los 4 artículos añadidos mediante búsqueda manual. La revisión se basa en la comparación de 9 estudios de cohortes para analizar la incidencia de la progresión de LPO a COCE. Conclusiones: Debemos considerar el LPO como una enfermedad potencialmente maligna (0.5-1%) siendo el sexo femenino, el tipo erosivo y la localización en la lengua factores de riesgo para la transformación a COCE. Teniendo en cuenta el riesgo de malignización es esencial el control de las lesiones para poder realizar un diagnóstico precoz

Introduction: Lichen planus (LP) is a systemic autoimmune disease of unknown etiology and chronic course that can result in the development of oral squamous cell carcinoma (OECD). Objective: To evaluate the evidence related to the malignization of the LPO as well as the clinical characteristics and the incidence of the lesions that can develop a COCE. Material and methods: The MEDLINE PubMed database of the last 5 years (2013-2018) was reviewed using the following keywords: "malignant transformation of oral lichen planus" and "squamous cell oral carcinoma". Results: 48 articles were obtained, after reading the summaries and applying our inclusion and exclusion criteria, we obtained a total of 19 articles, in addition to the 4 articles added by manual search. The review is based on the comparison of 9 cohort studies to analyze the incidence of progression from LPO to COCE. Conclusions: We should consider the LPO as a potentially malignant disease (0.5-1%), being the female sex, the erosive type and the localization in the tongue as a risk factors for the transformation to COCE. Taking into account the risk of malignancy, it is essential to control the lesions in order to make an early diagnosis

Inequalities in health? An update on the effect of social deprivation for patients with breast cancer in South East Wales.

BACKGROUND: Breast cancer, historically a disease of more affluent women, has increased in incidence for women from areas of greater social deprivation, yet prognosis is worse for these women. This study identifies differences in presentation, treatment and prognostic factors between the socioeconomic groups. METHODS: Patient data obtained from the prospectively maintained Welsh national Cancer Network Information System Cymru, for an 11-year period, were categorised according to Welsh Index of Multiple Deprivation quintiles. Quintiles were compared for differences in variables relating to patient characteristics, detection of cancer, tumour biology and treatment. RESULTS: 1570 patients were included. Analysis showed that in the more socially deprived quintiles, there are proportionally fewer women being diagnosed through the NHS breast cancer screening programme and as a consequence greater numbers of women from poorer areas being diagnosed outwith the screening age parameters. Screen detection is strongly associated with better prognosis in terms of Nottingham Prognostic Index. Similarly, increasing levels of social deprivation are associated with higher incidence of oestrogen receptor negative and triple negative tumours, both features associated with a shorter disease free and overall survival. Other variables of tumour biology, rates and type of surgical and adjuvant treatment were similar across social deprivation quintiles. CONCLUSION: There is a trend of reduced early detection of breast cancer in South East Wales in those patients living in areas of higher social deprivation. Given that there is equity in access to treatment within NHS, which is free for patients at the point of care, further study is warranted to address this existing disparity. Population cancer surveillance will need to inform both public health and NHS service responses, to continue to achieve improvements. Health trends may yet alter depending on current and future shifts in governmental health policy.

Comparison of liposomal drug formulations for transdermal iontophoretic drug delivery.

This study was aimed to evaluate the in vitro transdermal direct/pulsed current iontophoretic delivery of an amphiphilic model compound from various lipid vesicle-encapsulated formulations compared to free-drug formulation. Conventional, pegylated, ultradeformable liposomes (transfersomes) and ethosomes loaded with a negatively charged drug diclofenac sodium (DS) were prepared and characterized. All the liposomes possessed an average size of ≈100-150nm and negative zeta potential. No changes in colloidal stability were detected after 8h incubation of any vesicle formulation under constant or pulsed iontophoretic current. DS was released from all the liposome formulations with a similar, limited rate (≈50% in 24h), leading therefore to significantly lower transdermal fluxes across full-thickness porcine skin compared to the respective free drug formulation. From the tested lipid vesicle formulations, the transfersomes resulted in the highest passive flux and the ethosomes in the highest iontophoretic flux under direct constant current treatment. Higher negative surface charge of the vesicle led to better transport efficiency due to the higher mobility of the drug carrier under electric field. Pulsed current iontophoresis had no advantage over constant current treatment in combination with any type of lipid vesicular nanocarriers, in contrast to what has been described earlier with drug-loaded polymeric nanocarriers.

Predicting patterns of long-term adaptation and extinction with population genetics.

Population genetics struggles to model extinction; standard models track the relative rather than absolute fitness of genotypes, while the exceptions describe only the short-term transition from imminent doom to evolutionary rescue. But extinction can result from failure to adapt not only to catastrophes, but also to a backlog of environmental challenges. We model long-term adaptation to long series of small challenges, where fitter populations reach higher population sizes. The population's long-term fitness dynamic is well approximated by a simple stochastic Markov chain model. Long-term persistence occurs when the rate of adaptation exceeds the rate of environmental deterioration for some genotypes. Long-term persistence times are consistent with typical fossil species persistence times of several million years. Immediately preceding extinction, fitness declines rapidly, appearing as though a catastrophe disrupted a stably established population, even though gradual evolutionary processes are responsible. New populations go through an establishment phase where, despite being demographically viable, their extinction risk is elevated. Should the population survive long enough, extinction risk later becomes constant over time.

Transdermal iontophoresis of flufenamic acid loaded PLGA nanoparticles.

The objective of this study was to test in vitro a drug delivery system that combines nanoencapsulation and iontophoresis for the transdermal delivery of lipophilic model drug using poly(lactic-co-glycolic acid) (PLGA) as the carrier polymer. Negatively charged fluorescent nanoparticles loaded with negatively charged flufenamic acid were prepared. The colloidal properties of the particles were stable under iontophoretic current (constant, pulsed and alternating) profiles and in contact with skin barrier. The release of the drug from the particles was not affected by iontophoresis and remained always limited (≈50%), leading to significantly lower transdermal fluxes across human epidermis and full thickness porcine skin compared to respective free drug formulation. From nanoparticles, pulsed current profile resulted in comparable or higher fluxes compared to constant current profile although fluorescence imaging was not able to confirm deeper distribution of nanoparticles in skin. Based on our results, there is no clear advantage with respect to drug permeation from nanoencapsulating flufenamic acid into PLGA nanoparticles compared to free drug formulation, either in passive or iontophoretic delivery regimens. However, pulsed current iontophoresis could be an effective alternative instead of traditional constant current iontophoresis to enhance transdermal permeation of drugs from nanoencapsulated formulations.

Carboxylated nanodiamonds inhibit γ-irradiation damage of human red blood cells.

Nanodiamonds when carboxylated (cNDs) act as reducing agents and hence could limit oxidative damage in biological systems. Gamma (γ)-irradiation of whole blood or its components is required in immunocompetent patients to prevent transfusion-associated graft versus host disease (TA-GVHD). However, γ-irradiation of blood also deoxygenates red blood cells (RBCs) and induces oxidative damage, including abnormalities in cellular membranes and hemolysis. Using atomic force microscopy (AFM) and Raman spectroscopy, we examined the effect of cNDs on γ-irradiation mediated deoxygenation and morphological damage of RBCs. γ-Radiation induced several morphological phenotypes, including stomatocytes, codocytes and echinocytes. While stomatocytes and codocytes are reversibly damaged RBCs, echinocytes are irreversibly damaged. AFM images show significantly fewer echinocytes among cND-treated γ-irradiated RBCs. The Raman spectra of γ-irradiated RBCs had more oxygenated hemoglobin patterns when cND-treated, resembling those of normal, non-irradiated RBCs, compared to the non-cND-treated RBCs. cND inhibited hemoglobin deoxygenation and morphological damage, possibly by neutralizing the free radicals generated during γ-irradiation. Thus cNDs have the therapeutic potential to preserve the quality of stored blood following γ-irradiation.

Super-resolution microscopy as a potential approach to diagnosis of platelet granule disorders.

BACKGROUND: Many platelet functions are dependent on bioactive molecules released from their granules. Deficiencies of these granules in number, shape or content are associated with bleeding. The small size of these granules is such that imaging them for diagnosis has traditionally required electron microscopy. However, recently developed super-resolution microscopes provide sufficient spatial resolution to effectively image platelet granules. When combined with automated image analysis, these methods provide a quantitative, unbiased, rapidly acquired dataset that can readily and reliably reveal differences in platelet granules between individuals. OBJECTIVE: To demonstrate the ability of structured illumination microscopy (SIM) to efficiently differentiate between healthy volunteers and three patients with Hermansky-Pudlak syndrome. METHODS: Blood samples were taken from three patients with Hermansky-Pudlak syndrome and seven controls. Patients 1-3 have gene defects in HPS1, HPS6 and HPS5, respectively; all controls were healthy volunteers. Platelet-rich plasma was isolated from blood and the platelets fixed, stained for CD63 and processed for analysis by immunofluorescence microscopy, using a custom-built SIM microscope. RESULTS: SIM can successfully resolve CD63-positive structures in fixed platelets. A determination of the number of CD63-positive structures per platelet allowed us to conclude that each patient was significantly different from all of the controls with 99% confidence. CONCLUSIONS: A super-resolution imaging approach is effective and rapid in objectively differentiating between patients with a platelet bleeding disorder and healthy volunteers. CD63 is a useful marker for predicting Hermansky-Pudlak syndrome and could be used in the diagnosis of patients suspected of other platelet granule disorders.

Safety and efficacy of factor XI (FXI) concentrate use in patients with FXI deficiency: a single-centre experience of 19 years.

AIM: Factor XI (FXI) concentrate is a pooled human plasma-derived factor concentrate used as replacement therapy for patients with FXI deficiency, which provides a predictable response and consistent haemostatic cover in emergency or elective situations. It has previously been implicated in adverse events such as thrombosis and inhibitor formation, with rare case reports of fatal incidents. We sought to establish the incidence of such complications in a retrospective case series between 1994 and 2012 at the Haemophilia Comprehensive Care Centre at Royal Free Hospital, London, UK. METHODS: Patients who received FXI concentrate had their medical records reviewed to extract information and specific adverse events recorded such as failure of treatment with further bleeding, suspected viral transfusion transmitted infection (TTI), thrombosis or inhibitor formation. RESULTS: Eighty-six patients received 242 treatment episodes of FXI concentrate. Ninety percent of treatment episodes were covered with BPL FXI concentrate and 10% with LFB Hemoleven. Twelve (5%) adverse events were recorded, with eight (3.3%) of all treatment episodes were related to persistent bleeding postconcentrate infusion and there were 4 (1.7%) non-bleeding adverse events. No viral TTIs were identified. There were two recorded inhibitors, one thrombotic event (central retinal artery occlusion) and one transfusion reaction. No patient suffering an adverse event resulted in long-term morbidity. CONCLUSION: Our experience of FXI concentrate use demonstrates infrequent minor adverse events related to its administration and is a safe product to use.

Presentation and management of acute coronary syndromes among adult persons with haemophilia: results of an international, retrospective, 10-year survey.

Sparse data are available on presentation and management of acute coronary syndromes (ACS), including unstable angina and non-ST- and ST-elevation myocardial infarction, among persons with haemophilia (PWH). The aim of this study was to determine demographics, bleeding disorder characteristics, cardiovascular risk factors (CRFs), interventions, haemostatic protocol, revascularization outcomes and complications among PWH with ACS. Members of an international consortium comprising >2000 adult PWH retrospectively completed case report forms for episodes of ACS in a >10-year follow-up period (2003-2013). Twenty ACS episodes occurred among 19 patients [rate, 0.8% (95% CI 0.4, 1.2)]. Seven patients (37%) were aged <50 years; 10 (53%) had ≥3 CRFs. In 5/20 episodes (25%), the initial ACS management protocol was altered because of the bleeding disorder. None of the eight patients with severe haemophilia underwent coronary artery bypass grafting (CABG), compared with 54.5% of patients with non-severe disease (P = 0.02). Revascularization with percutaneous coronary intervention (PCI) or CABG was rated successful in 13/13 cases, with no excessive bleeding during initial management. During chronic exposure to antiplatelet agents, secondary haemophilia prophylaxis was more prevalent in patients with severe haemophilia compared with non-severe haemophilia (85.7% vs. 30%, P = 0.05). No ACS-related deaths occurred during initial management, but one patient with severe haemophilia A died of undetermined cause 36 months after the ACS event while on aspirin therapy. ACS occurs even among relatively younger PWH, typically in association with multiple CRFs. Revascularization with PCI/CABG is feasible, and antiplatelet agents plus secondary prophylaxis appears to be well tolerated in selected PWH with ACS.

Evaluación de la presencia de marcadores serológicos asociados al grado de avance de la cardiomiopatía chagásica crónica: estudio transversal / Evaluation of the presence of serological markers associated to progress of chronic chagasic cardiomyopathy: cross-sectional study

INTRODUCCIÓN El seguimiento de la progresión de la enfermedad cardíaca en la cardiomiopatía chagásica crónica podría mejorarse si se utilizaran marcadores serológicos con capacidad para detectar precozmente los primeros signos de afectación cardíaca. OBJETIVOS Estudiar los marcadores PAP-1, GAL3, sST2, endotelina 1, IL-6, IL-10, TNFα, IFNγ y los anticuerpos contra JL7, así como la asociación entre sus niveles y el grado de avance de la cardiomiopatía. MÉTODOS Se analizaron muestras de pacientes que asistían al Hospital Ramos Mejía, clasificados por su grado de afectación cardíaca y por el aspecto funcional. Para cada marcador se determinó su nivel por ELISA, y luego se compararon los grupos aplicando el test de Kruskal-Wallis. Para el estudio de los anticuerpos contra JL7, se analizaron las variantes de la proteína en los diferentes linajes del parásito y se ensayaron por ELISA los péptidos que representan estas variaciones. RESULTADOS La mediana para PAP-1 fue: 2,7 en asintomáticos, 7,3 en CARD1/2 y 26,95 ng/ ml en CARD 3/4, con diferencias significativas entre todos los grupos (p<0,001 entre CARD 3/4 y los otros dos grupos y p<0,05 entre CARD 1/2 y asintomáticos). En el caso de IL-6, el valor de la mediana fue de 8,2 pg/ml para los pacientes sin cardiomiopatía demostrable, mientras que para los asintomáticos/CARD 1 fue de 13,50 pg/ml y para los CARD 2/3, de 22,05 pg/ml. Sólo cuando se agruparon a los pacientes con sintomatología cardíaca y se efectuó la comparación con el grupo sin sintomatología, se vieron diferencias significativas entre ambos grupos (p<0,05, test de Mann Whitney). Para JL7 se encontraron péptidos con reactividad diferencial según la manifestación clínica del paciente JL7- 6/10/12 en asintomáticos, JL7-8 en cardiópatas y JL7-11 en pacientes con sintomatología gastrointestinal (p<0,01, test de Tukey). DISCUSIÓN Sólo se encontró una asociación entre los valores de los marcadores y el grado de compromiso cardíaco para PAP-1 e IL-6. También se hallaron variantes de JL7 asociadas a cierta manifestación clínica.

An interactive mutation database for human coagulation factor IX provides novel insights into the phenotypes and genetics of hemophilia B.

BACKGROUND: Factor IX (FIX) is important in the coagulation cascade, being activated to FIXa on cleavage. Defects in the human F9 gene frequently lead to hemophilia B. OBJECTIVE: To assess 1113 unique F9 mutations corresponding to 3721 patient entries in a new and up-to-date interactive web database alongside the FIXa protein structure. METHODS: The mutations database was built using MySQL and structural analyses were based on a homology model for the human FIXa structure based on closely-related crystal structures. RESULTS: Mutations have been found in 336 (73%) out of 461 residues in FIX. There were 812 unique point mutations, 182 deletions, 54 polymorphisms, 39 insertions and 26 others that together comprise a total of 1113 unique variants. The 64 unique mild severity mutations in the mature protein with known circulating protein phenotypes include 15 (23%) quantitative type I mutations and 41 (64%) predominantly qualitative type II mutations. Inhibitors were described in 59 reports (1.6%) corresponding to 25 unique mutations. CONCLUSION: The interactive database provides insights into mechanisms of hemophilia B. Type II mutations are deduced to disrupt predominantly those structural regions involved with functional interactions. The interactive features of the database will assist in making judgments about patient management.

Above- and below-ground methane fluxes and methanotrophic activity in a landfill-cover soil.

Landfills are a major anthropogenic source of the greenhouse gas methane (CH(4)). However, much of the CH(4) produced during the anaerobic degradation of organic waste is consumed by methanotrophic microorganisms during passage through the landfill-cover soil. On a section of a closed landfill near Liestal, Switzerland, we performed experiments to compare CH(4) fluxes obtained by different methods at or above the cover-soil surface with below-ground fluxes, and to link methanotrophic activity to estimates of CH(4) ingress (loading) from the waste body at selected locations. Fluxes of CH(4) into or out of the cover soil were quantified by eddy-covariance and static flux-chamber measurements. In addition, CH(4) concentrations at the soil surface were monitored using a field-portable FID detector. Near-surface CH(4) fluxes and CH(4) loading were estimated from soil-gas concentration profiles in conjunction with radon measurements, and gas push-pull tests (GPPTs) were performed to quantify rates of microbial CH(4) oxidation. Eddy-covariance measurements yielded by far the largest and probably most representative estimates of overall CH(4) emissions from the test section (daily mean up to ∼91,500µmolm(-2)d(-1)), whereas flux-chamber measurements and CH(4) concentration profiles indicated that at the majority of locations the cover soil was a net sink for atmospheric CH(4) (uptake up to -380µmolm(-2)d(-1)) during the experimental period. Methane concentration profiles also indicated strong variability in CH(4) loading over short distances in the cover soil, while potential methanotrophic activity derived from GPPTs was high (v(max)∼13mmolL(-1)(soil air)h(-1)) at a location with substantial CH(4) loading. Our results provide a basis to assess spatial and temporal variability of CH(4) dynamics in the complex terrain of a landfill-cover soil.

The role of recombinant factor VIIa for obstetric block in women with severe factor XI deficiency.

Regional blocks like spinal, epidural and combined spinal epidural (CSE) are relatively contraindicated in individuals with bleeding disorders. Consequently pregnant women with severe factor XI (FXI) deficiency are often denied this option during labour and caesarean section. We describe three women with severe FXI deficiency in whom regional block was performed with low-dose recombinant factor VIIa (rFVIIa) for their operative procedures during delivery. All women achieved haemostasis and had uncomplicated regional block, delivery and surgical procedures. The point of care device--rotational thromboelastometry (ROTEM) was used to monitor the patients' coagulation and determine the dose of rFVIIa to achieve in vitro haemostasis in these women that was then subsequently used in vivo. Low-dose rFVIIa seems to be effective and safe in the management of delivery and enables provision of regional blocks in women with severe FXI deficiency.