An artificial intelligence-generated model predicts 90-day survival in alcohol-associated hepatitis: A global cohort study.

BACKGROUND AND AIMS: Alcohol-associated hepatitis (AH) poses significant short-term mortality. Existing prognostic models lack precision for 90-day mortality. Utilizing artificial intelligence in a global cohort, we sought to derive and validate an enhanced prognostic model. APPROACH AND RESULTS: The Global AlcHep initiative, a retrospective study across 23 centers in 12 countries, enrolled patients with AH per National Institute for Alcohol Abuse and Alcoholism criteria. Centers were partitioned into derivation (11 centers, 860 patients) and validation cohorts (12 centers, 859 patients). Focusing on 30 and 90-day postadmission mortality, 3 artificial intelligence algorithms (Random Forest, Gradient Boosting Machines, and eXtreme Gradient Boosting) informed an ensemble model, subsequently refined through Bayesian updating, integrating the derivation cohort's average 90-day mortality with each center's approximate mortality rate to produce posttest probabilities. The ALCoholic Hepatitis Artificial INtelligence Ensemble score integrated age, gender, cirrhosis, and 9 laboratory values, with center-specific mortality rates. Mortality was 18.7% (30 d) and 27.9% (90 d) in the derivation cohort versus 21.7% and 32.5% in the validation cohort. Validation cohort 30 and 90-day AUCs were 0.811 (0.779-0.844) and 0.799 (0.769-0.830), significantly surpassing legacy models like Maddrey's Discriminant Function, Model for End-Stage Liver Disease variations, age-serum bilirubin-international normalized ratio-serum Creatinine score, Glasgow, and modified Glasgow Scores ( p < 0.001). ALCoholic Hepatitis Artificial INtelligence Ensemble score also showcased superior calibration against MELD and its variants. Steroid use improved 30-day survival for those with an ALCoholic Hepatitis Artificial INtelligence Ensemble score > 0.20 in both derivation and validation cohorts. CONCLUSIONS: Harnessing artificial intelligence within a global consortium, we pioneered a scoring system excelling over traditional models for 30 and 90-day AH mortality predictions. Beneficial for clinical trials, steroid therapy, and transplant indications, it's accessible at: https://aihepatology.shinyapps.io/ALCHAIN/ .

Is It Safe to Omit the 1-week Post-operative Examination after Uncomplicated Phacoemulsification?

SIGNIFICANCE: The incidence of cataract surgery is increasing, accounting for a large percentage of eye care expenses. Scientific evidence supporting the medical necessity of the traditional post-operative schedule is lacking. Further studies are needed to optimize post-operative care to reduce the burden on patients and medical providers. PURPOSE: This study aimed to study the rate of complication 1 week after uncomplicated phacoemulsification to determine if the 1-week post-operative examination can be safely omitted. METHODS: A retrospective record review was conducted on all consecutive patients who had uncomplicated phacoemulsification between February 1, 2019, and February 1, 2020, at a clinic in an urban setting with a predominantly Black and African American patient population. Subjects were included if they had no complications during the 1-day post-operative examination. Complications at the 1-week and 1-month post-operative examination were recorded and analyzed. RESULTS: Omitting the 1-week post-operative examination would result in missed complications in 4.48 to 15.97% of patients and failure to make unexpected management changes in 1.78 to 13.84% of patients. CONCLUSIONS: The results of this study do not support omitting the 1-week post-operative examination after uncomplicated phacoemulsification. Further studies are needed to determine whether telemedicine can be safely substituted for post-operative examinations.

SARS-CoV-2 self-testing in Peru: a cross-sectional survey of values and attitudes of the general population.

OBJECTIVES: To assess the values of and attitudes towards the use of rapid SARS-CoV-2 antigen-detection tests for self-testing in a rural and an urban area in Peru. DESIGN: Cross-sectional, street-based population survey. SETTING: A series of over 400 randomly selected street points in Valle del Mantaro and in Lima. PARTICIPANTS: 438 respondents (203 female) participated. They were all older than 17 years and provided informed consent for participation. INTERVENTION: All respondents answered on the spot, a 35-item questionnaire developed in KoboToolbox. PRIMARY AND SECONDARY OUTCOME MEASURES: Primary outcomes of interest were: likelihood to use a SARS-CoV-2 self-test; willingness to pay for a SARS-CoV-2 self-test and likelihood to comply with recommended actions following a positive SARS-CoV-2 self-test result. Bivariate analyses and Poisson regression (PR) analyses were performed to identify significant associations between dependent variables and independent variables pertaining to respondents' characteristics, risk perception and previous experiences with conventional COVID-19 testing. RESULTS: Of the 438 respondents, 51.49% had previous experience with conventional COVID-19 testing; 20.37% had COVID-19 disease; 86.96% accepted the idea of SARS-CoV-2 self-testing; and, 78.95% would be likely to use it if needed. Almost all (94.75%) would pay for a self-testing device (mean acceptable payment: US$10.4) if it was not provided free of charge by health authorities. Overall, 93.12%, 86.93% and 85.32% would self-isolate, report the results and warn their contacts, respectively. Being a female (adjusted PR 1.05, 95% CI 1.00 to 1.09, p<0.018), having completed secondary education (adjusted PR 1.18, 95% CI 1.02 to 1.37, p<0.024) and expressing likelihood to use self-testing (adjusted PR 1.08, 95% CI 1.01 to 1.16, p<0.0.24) could be predictors of willingness to pay for a self-test. CONCLUSIONS: Self-testing is perceived as an acceptable approach. Health authorities in Peru should facilitate access to this approach to complement healthcare facilities-led testing efforts for COVID-19. Future research is necessary to understand the impact of self-testing in case detection and pandemic control.

MELD 3.0 adequately predicts mortality and renal replacement therapy requirements in patients with alcohol-associated hepatitis.

Background & Aims: Model for End-Stage Liver Disease (MELD) score better predicts mortality in alcohol-associated hepatitis (AH) but could underestimate severity in women and malnourished patients. Using a global cohort, we assessed the ability of the MELD 3.0 score to predict short-term mortality in AH. Methods: This was a retrospective cohort study of patients admitted to hospital with AH from 2009 to 2019. The main outcome was all-cause 30-day mortality. We compared the AUC using DeLong's method and also performed a time-dependent AUC with competing risks analysis. Results: A total of 2,124 patients were included from 28 centres from 10 countries on three continents (median age 47.2 ± 11.2 years, 29.9% women, 71.3% with underlying cirrhosis). The median MELD 3.0 score at admission was 25 (20-33), with an estimated survival of 73.7% at 30 days. The MELD 3.0 score had a better performance in predicting 30-day mortality (AUC:0.761, 95%CI:0.732-0.791) compared with MELD sodium (MELD-Na; AUC: 0.744, 95% CI: 0.713-0.775; p = 0.042) and Maddrey's discriminant function (mDF) (AUC: 0.724, 95% CI: 0.691-0.757; p = 0.013). However, MELD 3.0 did not perform better than traditional MELD (AUC: 0.753, 95% CI: 0.723-0.783; p = 0.300) and Age-Bilirubin-International Normalised Ratio-Creatinine (ABIC) (AUC:0.757, 95% CI: 0.727-0.788; p = 0.765). These results were consistent in competing-risk analysis, where MELD 3.0 (AUC: 0.757, 95% CI: 0.724-0.790) predicted better 30-day mortality compared with MELD-Na (AUC: 0.739, 95% CI: 0.708-0.770; p = 0.028) and mDF (AUC:0.717, 95% CI: 0.687-0.748; p = 0.042). The MELD 3.0 score was significantly better in predicting renal replacement therapy requirements during admission compared with the other scores (AUC: 0.844, 95% CI: 0.805-0.883). Conclusions: MELD 3.0 demonstrated better performance compared with MELD-Na and mDF in predicting 30-day and 90-day mortality, and was the best predictor of renal replacement therapy requirements during admission for AH. However, further prospective studies are needed to validate its extensive use in AH. Impact and implications: Severe AH has high short-term mortality. The establishment of treatments and liver transplantation depends on mortality prediction. We evaluated the performance of the new MELD 3.0 score to predict short-term mortality in AH in a large global cohort. MELD 3.0 performed better in predicting 30- and 90-day mortality compared with MELD-Na and mDF, but was similar to MELD and ABIC scores. MELD 3.0 was the best predictor of renal replacement therapy requirements. Thus, further prospective studies are needed to support the wide use of MELD 3.0 in AH.

Ruxolitinib as a treatment strategy for SARS-CoV-2 pneumonia: clinical experience in a real-world setting.

INTRODUCTION: Severe acute respiratory syndrome-related coronavirus (SARS-CoV-2) infection is characterised by a viral phase and a severe pro-inflammatory phase. The inhibition of the JAK/STAT pathway limits the pro-inflammatory state in moderate to severe COVID-19. METHODOLOGY: We analysed the data obtained by an observational cohort of patients with SARS-CoV-2 pneumonia treated with ruxolitinib in 22 hospitals of Mexico. The applied dose was determined based on physician's criteria. The benefit of ruxolitinib was evaluated using the 8-points ordinal scale developed by the NIH in the ACTT1 trial. Duration of hospital stay, changes in pro-inflammatory laboratory values, mortality, and toxicity were also measured. RESULTS: A total of 287 patients were reported at 22 sites in Mexico from March to June 2020; 80.8% received ruxolitinib 5 mg BID and 19.16% received ruxolitinib 10 mg BID plus standard of care. At beginning of treatment, 223 patients were on oxygen support and 59 on invasive ventilation. The percentage of patients on invasive ventilation was 53% in the 10 mg and 13% in the 5 mg cohort. A statistically significant improvement measured as a reduction by 2 points on the 8-point ordinal scale was described (baseline 5.39 ± 0.93, final 3.67± 2.98, p = 0.0001). There were 74 deaths. Serious adverse events were presented in 6.9% of the patients. CONCLUSIONS: Ruxolitinib appears to be safe in COVID-19 patients, with clinical benefits observed in terms of decrease in the 8-point ordinal scale and pro-inflammatory state. Further studies must be done to ensure efficacy against mortality.

Feasibility of Noninvasive Positive Pressure Ventilation in the Treatment of Oxygen-Dependent COVID-19 Patients in Peru.

Intensive care is expensive, and availability is limited. Low- and middle-income countries in particular have struggled to cope with the large influx of critically ill patients during the COVID-19 pandemic. Noninvasive respiratory support devices delivering continuous positive airways pressure (CPAP) require less resource and staff expertise compared with invasive mechanical ventilators and can be routinely used outside of intensive care units. This study assessed the use of the UCL-Ventura Wayrachi CPAP device in hospitalized patients with COVID-19 in Peru. A secondary analysis of data collected for a feasibility study commissioned by the Peruvian Ministry of Health was conducted. Data were collected from three hospitals, including patient demographics, clinical data, and outcomes. Forty-five patients were enrolled from July 16 to September 1, 2020. Eight patients (18%) were intolerant of the CPAP mask. Of the remainder, 18 (48.7%) improved and were discharged from hospital after 6 days. Eight (21.6%) died while on CPAP and 11 (29.7%) were eventually intubated, of whom two died. In total, 27 (60%) survived to hospital discharge. Participating physicians noted the device was easy to use and provided patient benefit, though voiced concerns about the strain on hospital oxygen supplies. In conclusion, the UCL Ventura Wayrachi CPAP device proved feasible in COVID-19 patients in Peru, and offered a bridging therapy for patients who required a ventilator when none were available.

Identification of optimal therapeutic window for steroid use in severe alcohol-associated hepatitis: A worldwide study.

BACKGROUND & AIMS: Corticosteroids are the only effective therapy for severe alcohol-associated hepatitis (AH), defined by a model for end-stage liver disease (MELD) score >20. However, there are patients who may be too sick to benefit from therapy. Herein, we aimed to identify the range of MELD scores within which steroids are effective for AH. METHODS: We performed a retrospective, international multicenter cohort study across 4 continents, including 3,380 adults with a clinical and/or histological diagnosis of AH. The main outcome was mortality at 30 days. We used a discrete-time survival analysis model, and MELD cut-offs were established using the transform-the-endpoints method. RESULTS: In our cohort, median age was 49 (40-56) years, 76.5% were male, and 79% had underlying cirrhosis. Median MELD at admission was 24 (19-29). Survival was 88% (87-89) at 30 days, 77% (76-78) at 90 days, and 72% (72-74) at 180 days. A total of 1,225 patients received corticosteroids. In an adjusted-survival-model, corticosteroid use decreased 30-day mortality by 41% (hazard ratio [HR] 0.59; 0.47-0.74; p <0.001). Steroids only improved survival in patients with MELD scores between 21 (HR 0.61; 0.39-0.95; p = 0.027) and 51 (HR 0.72; 0.52-0.99; p = 0.041). The maximum effect of corticosteroid treatment (21-30% survival benefit) was observed with MELD scores between 25 (HR 0.58; 0.42-0.77; p <0.001) and 39 (HR 0.57; 0.41-0.79; p <0.001). No corticosteroid benefit was seen in patients with MELD >51. The type of corticosteroids used (prednisone, prednisolone, or methylprednisolone) was not associated with survival benefit (p = 0.247). CONCLUSION: Corticosteroids improve 30-day survival only among patients with severe AH, especially with MELD scores between 25 and 39. LAY SUMMARY: Alcohol-associated hepatitis is a condition where the liver is severely inflamed as a result of excess alcohol use. It is associated with high mortality and it is not clear whether the most commonly used treatments (corticosteroids) are effective, particularly in patients with very severe liver disease. In this worldwide study, the use of corticosteroids was associated with increased 30-day, but not 90- or 180-day, survival. The maximal benefit was observed in patients with an MELD score (a marker of severity of liver disease; higher scores signify worse disease) between 25-39. However, this benefit was lost in patients with the most severe liver disease (MELD score higher than 51).

Hiperprocalcitonemia como pronóstico clínico en pacientes sépticos de los centros hospitalarios privados de la ciudad de Cuenca ­ Ecuador / Hyperprocalcitonemia as a clinical prognosis in septic patients of private hospitals in the city of Cuenca­Ecuador

La procalcitonina, reactante de fase aguda, permite establecer el estado de los pacientes con diagnóstico de sepsis, brindando la posibilidad de asociarlo con su pronóstico. El objetivo de este estudio fue el determinar el pronóstico clínico de la hiperprocalcitonemia en pacientes sépticos de los centros hospitalarios privados de Cuenca­Ecuador. El estudio fue analítico de corte transversal, la muestra fueron 207 pacientes. Los datos se analizaron con el programa estadístico SPSS (25,0); el análisis se realizó mediante estadística descriptiva; la asociación mediante Odds Ratio (OR), intervalo de confianza (95%), considerando valores estadísticamente significativos con p <0,05. La prevalencia de hiperprocalcitonemia severa fue 63,29%, la media de edad 67,25±19,07 años; predominó el sexo masculino 57% y la etnia mestiza. Se evidenció asociación estadística entre hiperprocalcitonemia severa y mayor estancia hospitalaria (>15 días) OR: 2,41 (IC 95% 1,11-5,19 p: 0,015); de igual manera con la mortalidad intrahospitalaria OR: 9,37 (IC 95% 4,31-20,37 p: <0,000). Se determinó asociación, mas no significancia estadística con la presencia de comorbilidades OR: 1,35 (IC 95% 0,69-2,64 p: 0,243). Se evidenció hiperprocalcitonemia severa en casi 2/3 de los pacientes, y existió asociación con aumento de mortalidad y estancia hospitalaria.

Procalcitonin, an acute phase's reactant, enables to establish sepsis-diagnosis'-patients status, bringing the possibility of associate it with its prognosis. The aim of this study was to determine the clinical prognosis of hyperprocalcitonemia in septical patients of private hospital centers in Cuenca­Ecuador. The study was cross-sectional, the sample were 207 patients. Data was analyzed with SPSS statistical program (25.0); analysis was done through descriptive statistic; association through Odds Ratio (OR), confidence interval (95%), considering statiscally significant values with p <0.05. Severe hyperprocalcitonemia prevalence was 63.29%, average age 67.25±19.07 years old; male sex prevailed 57% and half-blood ethnic group. A statistical association between severe hyperprocalcitonemia and longer hospital stay (≥15 days) was shown OR: 2.41 (CI 95% 1.11­5.19 p: 0.015); likewise, with in-hospital mortality OR: 9.37 (CI 95% 4.31­20.37 p: <0.000). Association was determined, but statistical significance with presence of comorbidities was not OR: 1.35 (CI 95% 0.69­2.64 p:0.243). Severe hyperprocalcitonemia was shown in almost 2/3 of patients, and there was an association with mortality increase and hospital stay.

Physiognomic responses of páramo tussock grass to time since fire in northern Ecuador / Respuestas fisionómicas de las macollas de gramínea del páramo posteriores a incendios en el norte del Ecuador

Abstract Ecologically-sound management plans for high-altitude grasslands of the Andes depend on an understanding the responses of plants to fire, especially the dominant tussock grasses. This study considers physiognomic responses of tussock grass in 13 sites in northern Ecuador with a known fire history, with time since fire 0.5-10 y, and a control site which had not been burned for at least 40 y. At each site, we assessed vegetation height, basal cover of the tussocks, and the ratio of dead:live leaves in tussocks. We also measured light at ground level. As recovery time increased, tussock cover and number decreased, while tussock height increased. Light levels fell sharply below the tussock canopies as recovery took place, and dead leaves accumulated quickly, reaching 60 - 70% by just two years after fire. The modification of physical tussock structure is likely to influence a much wider ecosystem response to fire, and determines directly the fuel load for future fires. Despite these clear changes in tussock characteristics, they were too variable to be used as a reliable bioindicator of time since fire. However, a better understanding of the responses of tussock grasses to fire and particularly its impact on other species should become the focus of further attention in future.

Resumen Los planes de manejo ecológicamente sólidos para los pastizales de gran altura en los Andes dependen del entendimiento de las respuestas de las plantas al fuego, en especial las respuestas de los pastos dominantes. Este estudio considera las respuestas fisionómicas de pastos en 13 sitios en el norte del Ecuador con un historial de incendios conocido, con tiempo entre 0.5 y 10 años después del incendio, más un sitio control donde al menos durante 40 años no se había producido incendio. En cada sitio, evaluamos la altura de la vegetación, la cobertura basal de las macollas y la proporción de hojas muertas:vivas en las macollas. También, medimos la luz a nivel del suelo. A medida que aumentó el tiempo de recuperación, la cobertura y el número de macollas disminuyeron, mientras que la altura de las macollas aumentó. Los niveles de luz cayeron fuertemente debajo de las copas de las macollas durante la recuperación, y las hojas muertas se acumularon rápidamente, alcanzando 60 - 70% solo dos años después del incendio. Es probable que las modificaciones en la estructura física de las macollas influyen en una respuesta mucho más amplia del ecosistema al fuego, y determinen directamente la carga de combustible para futuros incendios. A pesar de que se observaron cambios claros en las características de las macollas, estas eran demasiado variables para ser consideradas como un bioindicador confiable del tiempo transcurrido después del incendio. Sin embargo, una mejor comprensión de las respuestas de las macollas al fuego y, en particular, el impacto de esas respuestas en otras especies debería ser el enfoque de mayor atención en el futuro.

Current and future suitable habitat areas for Nasuella olivacea (Gray, 1865) in Colombia and Ecuador and analysis of its distribution across different land uses.

Nasuella olivacea is an endemic mammal from the Andes of Ecuador and Colombia. Due to its rarity, aspects about its natural history, ecology and distribution patterns are not well known, therefore, research is needed to generate knowledge about this carnivore and a first step is studying suitable habitat areas. We performed Ecological Niche Models and applied future climate change scenarios (2.6 and 8.5 RCP) to determine the potential distribution of this mammal in Colombia and Ecuador, with current and future climate change conditions; furthermore, we analysed its distribution along several land covers. We found that N. olivacea is likely to be found in areas where no records have been reported previously; likewise, climate change conditions would increase suitable distribution areas. Concerning land cover, 73.4% of N. olivacea potential distribution was located outside Protected Areas (PA), 46.1% in Forests and 40.3% in Agricultural Lands. These findings highlight the need to further research understudied species, furthering our understanding about distribution trends and responses to changing climatic conditions, as well as informig future PA designing. These are essential tools for supporting wildlife conservation plans, being applicable for rare species whose biology and ecology remain unknown.

GABA promotes gastrin-releasing peptide secretion in NE/NE-like cells: Contribution to prostate cancer progression.

In prostate cancer (PCa), neuroendocrine cells (NE) have been associated with the progression of the disease due to the secretion of neuropeptides that are capable of diffusing and influence surrounding cells. The GABAergic system is enriched in NE-like cells, and contributes to PCa progression. Additionally, γ-aminobutyric acid (GABA) stimulates the secretion of gastrin-releasing peptide (GRP) in peripheral organs. For the first time, in this study we show the role of GABA and GABAB receptor 1 (GABBR1) expression in GRP secretion in NE-like prostate cancer cells. We demonstrated an increase in GRP levels in NE-like cell medium treated with GABAB receptor agonist. Moreover, the blocking of this receptor inhibited GABA-induced GRP secretion. The invasive potential of PC3 cells was enhanced by either GRP or conditioned medium of NE-like cells treated with GABA. Additionally, we confirmed a positive correlation between GABA and GRP levels in the serum of PCa patients with NE markers. Finally, using public available data sets, we found a negative correlation between GABBR1 and androgen receptor (AR) expression, as well as a strong positive correlation between GABBR1 and enolase 2. These results suggest that GABA via GABBR1 induces GRP secretion in NE like cells involved in PCa progression.

[IgG4-related disease: a case report]. / Enfermedad asociada a IgG4: reporte de un caso.

IgG4-related disease is an inflammatory condition characterized by high levels of IgG4. It affects salivary and lacrimal glands, pancreas, lymph nodes, lungs or kidney. The diagnosis is based on identifying a histological pattern with a dense lymphocyte and plasmacyte infiltration, focal fibrosis or phlebitis, finding more than 10 IgG4 positive cells per high power field and/or IgG4/IgG ratio in plasma higher than 40%. We present a patient with Mikulicz's disease who meets histological findings required for the diagnosis of IgG4 related disease.

La enfermedad relacionada con IgG4 es una condición fibroinflamatoria en la que existe elevación de IgG4, afección a nivel de glándulas salivares, lacrimales, páncreas, ganglios linfáticos y pulmón. Para su diagnóstico se requiere la identificación de un patrón histológico sugestivo que muestre infiltrado linfoplasmocitario denso, fibrosis focal o flebitis a nivel de una glándula, más de 10 células positivas para IgG4 por campo de gran aumento y relación de IgG4/IgG arriba de 40% en plasma. Describimos el caso de una paciente que presentó enfermedad de Mikulicz y cumplió con los datos histológicos para diagnóstico de enfermedad relacionada con IgG4.

A two-step synthetic strategy to obtain a water-soluble derivative of curcumin with improved antioxidant capacity and in vitro cytotoxicity in C6 glioma cells.

A novel water-soluble derivative of curcumin (Cur-[G-2]-OH) was designed and synthesized from accessible raw materials in only two steps with an overall yield of 80%. The modification of curcumin phenol groups with second-generation polyester dendrons (dendronization) as a strategy to achieve an optimal hydrophilic/hydrophobic balance allows the complete water solubilization of the new curcumin derivative (5mg/ml) at room temperature. The therapeutic potential of Cur-[G-2]-OH was investigated in terms of antioxidant capacity, intracellular uptake and cytotoxicity in both rat glioblastoma cells and normal human dermal fibroblasts. Although the phenolic groups of curcumin were locked by dendronization, Cur-[G-2]-OH exhibited antioxidant capacity in water that was even higher than curcumin in dimethylsulfoxide (DMSO). This compound showed a steady cellular uptake contrasted with curcumin, which has a saturation capture at high concentrations. Combined with improved stability, this property seems to allow the intracellular accumulation of Cur-[G-2]-OH. Furthermore, the new compound exhibited increased cytotoxicity in rat C6 glioma cells in a time- and concentration-dependent manner, whereas in normal human fibroblasts, its IC50 value was >600µM versus the IC50 of curcumin found between 100 and 200µM. Surprisingly, Cur-[G-2]-OH drives cell death of C6 cells by a different mechanism of apoptosis triggered by curcumin. Together, these results suggest that curcumin dendronization could promote molecular and cellular mechanisms that are different from those induced by curcumin, presumably due to structural factors and not only for improved water solubility.

Semaphorin 3C Released from a Biocompatible Hydrogel Guides and Promotes Axonal Growth of Rodent and Human Dopaminergic Neurons.

Cell therapy in experimental models of Parkinson's disease replaces the lost dopamine neurons (DAN), but we still need improved methods to guide dopaminergic axons (DAx) of grafted neurons to make proper connections. The protein Semaphorin 3C (Sema3C) attracts DAN axons and enhances their growth. In this work, we show that the hydrogel PuraMatrix, a self-assembling peptide-based matrix, incorporates Sema3C and releases it steadily during 4 weeks. We also tested if hydrogel-delivered Sema3C attracts DAx using a system of rat midbrain explants embedded in collagen gels. We show that Sema3C released by this hydrogel attracts DAx, in a similar way to pretectum, which is known to attract growing DAN axons. We assessed the effect of Sema3C on the growth of DAx using microfluidic devices. DAN from rat midbrain or those differentiated from human embryonic stem cells showed enhanced axonal extension when exposed to hydrogel-released Sema3C, similar to soluble Sema3C. Notably, DAN of human origin express the cognate Sema3C receptors, Neuropilin1 and Neuropilin2. These results show that PuraMatrix is able to incorporate and release Sema3C, and such delivery guides and promotes the axonal growth of DAN. This biocompatible hydrogel might be useful as a Sema3C carrier for in vivo studies in parkinsonian animal models.

Epilepsy and hippocampal neurodegeneration induced by glutamate decarboxylase inhibitors in awake rats.

Glutamic acid decarboxylase (GAD), the enzyme responsible for GABA synthesis, requires pyridoxal phosphate (PLP) as a cofactor. Thiosemicarbazide (TSC) and γ-glutamyl-hydrazone (PLPGH) inhibit the free PLP-dependent isoform (GAD65) activity after systemic administration, leading to epilepsy in mice and in young, but not in adult rats. However, the competitive GAD inhibitor 3-mercaptopropionic acid (MPA) induces convulsions in both immature and adult rats. In the present study we tested comparatively the epileptogenic and neurotoxic effects of PLPGH, TSC and MPA, administered by microdialysis in the hippocampus of adult awake rats. Cortical EEG and motor behavior were analyzed during the next 2h, and aspartate, glutamate and GABA were measured by HPLC in the microdialysis-collected fractions. Twenty-four hours after drug administration rats were fixed for histological analysis of the hippocampus. PLPGH or TSC did not affect the motor behavior, EEG or cellular morphology, although the extracellular concentration of GABA was decreased. In contrast, MPA produced intense wet-dog shakes, EEG epileptiform discharges, a >75% reduction of extracellular GABA levels and remarkable neurodegeneration of the CA1 region, with >80% neuronal loss. The systemic administration of the NMDA glutamate receptor antagonist MK-801 30 min before MPA did not prevent the MPA-induced epilepsy but significantly protected against its neurotoxic effect, reducing neuronal loss to <30%. We conclude that in adult awake rats, drugs acting on PLP availability have only a weak effect on GABA neurotransmission, whereas direct GAD inhibition produced by MPA induces hyperexcitation leading to epilepsy and hippocampal neurodegeneration. Because this degeneration was prevented by the blockade of NMDA receptors, we conclude that it is due to glutamate-mediated excitotoxicity consequent to disinhibition of the hippocampal excitatory circuits.

Association between AT1 and AT2 angiotensin II receptor expression with cell proliferation and angiogenesis in operable breast cancer.

Angiotensin II (ANGII) has been associated with vascular proliferation in tumor and non-tumor models through its receptors AT1 and AT2. Our objective was to determine AT1 and AT2 receptor expression in operable breast cancer and its association with tumor grade, vascular density, and cellular proliferation. Seventy-seven surgically malignant breast tumors with no distant metastasis were included, and 7 benign lesions were used as controls. AT1 and AT2 receptor expression was determined by RT-PCR and immunohistochemistry (IHC) in 68 out of the 77 malignant lesions and in the 7 benign lesions. AT1 and AT2 receptor expression was detected in 35.3 and 25 % of cases, in both RT-PCR and IHC. Tumors that express AT1 showed an increase in T3 stage (92.3 vs. 7.7 % p < 0.001), mitotic index (4 ± 1 vs 2 ± 1, p = 0.05), vascular density (15 ± 3 vs 8 ± 5, p = 0.05), and cellular proliferation (85 ± 18 vs 55 ± 10, p = 0.01) versus AT1-negative lesions. Non-differences between clinical-pathologic variables and AT2 expression were found. AT1 receptor expression was associated to enhance angiogenesis and cellular proliferation rate, but no relationship with AT2 was found. ANGII and its peptides might play a role in the development and pathophysiology of breast cancer, and this could be valuable in the in the development of targeted therapies.

Concise review: Generation of neurons from somatic cells of healthy individuals and neurological patients through induced pluripotency or direct conversion.

Access to healthy or diseased human neural tissue is a daunting task and represents a barrier for advancing our understanding about the cellular, genetic, and molecular mechanisms underlying neurogenesis and neurodegeneration. Reprogramming of somatic cells to pluripotency by transient expression of transcription factors was achieved a few years ago. Induced pluripotent stem cells (iPSC) from both healthy individuals and patients suffering from debilitating, life-threatening neurological diseases have been differentiated into several specific neuronal subtypes. An alternative emerging approach is the direct conversion of somatic cells (i.e., fibroblasts, blood cells, or glial cells) into neuron-like cells. However, to what extent neuronal direct conversion of diseased somatic cells can be achieved remains an open question. Optimization of current expansion and differentiation approaches is highly demanded to increase the differentiation efficiency of specific phenotypes of functional neurons from iPSCs or through somatic cell direct conversion. The realization of the full potential of iPSCs relies on the ability to precisely modify specific genome sequences. Genome editing technologies including zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeat/CAS9 RNA-guided nucleases have progressed very fast over the last years. The combination of genome-editing strategies and patient-specific iPSC biology will offer a unique platform for in vitro generation of diseased and corrected neural derivatives for personalized therapies, disease modeling and drug screening.

Detection of Epstein-Barr virus and genotyping based on EBNA2 protein in Mexican patients with hodgkin lymphoma: a comparative study in children and adults.

BACKGROUND: Epstein-Barr virus (EBV) is a member of the Herpesviridae family and is associated with Hodgkin lymphoma (HL). Isolates of EBV are classified according to sequence variation in the latency genes such as Epstein-Barr virus nuclear antigen (EBNA). EBNA2 contains the most divergent locus and is classified into type 1 and type 2 or EBNA2A and EBNA2B, respectively. We compared the frequency of EBV and the distribution of EBNA genotypes in Mexican children and adults with HL. PATIENTS AND METHODS: Lymph node biopsy specimens from children and adults with HL were embedded in paraffin. EBV was identified by LMP1 amplification and Epstein-Barr-encoded RNA EBER by in situ hybridization (ISH) and genotyped as EBNA2A or EBNA2B using nested polymerase chain reaction (PCR) and specific primers for the detection of subtype. RESULTS: Sixty-six samples were obtained from 3 hospitals-42 (63%) from children and 24 (37%) from adults with HL. Thirty-two of the 42 samples (76.1%) were positive for EBV in children and 16 of 24 (66.6%) samples were positive in adults (P = .41). In both children and adults, EBV was found more frequently in male patients. Thirty-four of 48 cases could be typed (70.8%). EBNA2A was found in 7/21 (33.3%) children and in 4/13 (30.8%) adults (P = 1.0), and EBNA2B was found in 10/21 (47.6%) children and in 9/13 (69.2%) adults (P = .22). A mix of subtypes was found in 4/21 (19%) children. CONCLUSION: EBV was found frequently in both children and adults with HL. EBNA2B was the most frequent subtype, and a high frequency of mixed subtypes was found in children.

[National guidelines of diagnosis and treatment of the non-Hodgkin lymphoma]. / Guías nacionales de diagnóstico y tratamiento de linfoma no Hodgkin.

Non-Hodgkin lymphoma comprises a heterogeneous group of haematological malignancies, classified according to their clinic, anatomic-pathological features and, lately, to their molecular biomarkers. Despite the therapeutic advances, nearly half of the patients will die because of this disease. The new diagnostic tools have been the cornerstone to design recent therapy targets, which must be included in the current treatment guidelines of this sort of neoplasms by means of clinical trials and evidence-based medicine. In the face of poor diagnoses devices in most of the Mexican hospitals, we recommend the present diagnose stratification, and treatment guidelines for non-Hodgkin lymphoma, based on evidence. They include the latest and most innovative therapeutic approaches, as well as specific recommendations for hospitals with limited framework and therapy resources.

Adjuvant immunotherapy of C6 glioma in rats with pertussis toxin.

PURPOSE: In spite of the recent advances in surgery and antitumor drugs, the brain tumors, like glioblastoma, have shown a poor prognosis. The aim of this study was to determine the effect of pertussis toxin (PTx) as immunomodulatory molecule on glial tumors induced by C6 glioma cells. METHODS: Given the pleiotropic effect of PTx on the immune system, we analyzed the effect of PTx on CD4+/CD25+/FoxP3+ (Treg) cells like as immunotherapeutic adjuvant. Thirty rats with a glial tumor of 1.5 cm in diameter were separated in two groups: the first group was treated with PTx and the second group was non-treated (controls). Tumoral volume was measured weekly; tumor, blood and spleen were taken for analysis of subpopulations of T cells, apoptotic index and cytokine contents, in both groups. RESULTS: We observed a significant decrease in tumor volume in the PTx group; this was associated with a decreased in the number of Treg cells, in both spleen and tumor. The percentage of apoptotic cells was increased as compared with that of controls. The production of proinflammatory cytokines was increased in mRNA for IL-6 as well as a small increase in the mRNA expression of perforin and granzime in tumors from rats treated with PTx. No changes were found in the mRNA expression of MCP-1 and MIP-1α. CONCLUSION: These results suggest that PTx could be an immunotherapeutic adjuvant in the integral therapy against glial tumors.