Discovery of Ligands for TRIM58, a Novel Tissue-Selective E3 Ligase.

Redirecting E3 ligases to neo-substrates, leading to their proteasomal disassembly, known as targeted protein degradation (TPD), has emerged as a promising alternative to traditional, occupancy-driven pharmacology. Although the field has expanded tremendously over the past years, the choice of E3 ligases remains limited, with an almost exclusive focus on CRBN and VHL. Here, we report the discovery of novel ligands to the PRY-SPRY domain of TRIM58, a RING ligase that is specifically expressed in erythroid precursor cells. A DSF screen, followed by validation using additional biophysical methods, led to the identification of TRIM58 ligand TRIM-473. A basic SAR around the chemotype was established by utilizing a competitive binding assay employing a short FP peptide probe derived from an endogenous TRIM58 substrate. The X-ray co-crystal structure of TRIM58 in complex with TRIM-473 gave insights into the binding mode and potential exit vectors for bifunctional degrader design.

Precisión de la calculadora de riesgo quirúrgico ACS NSQIP para predecir morbilidad y mortalidad en pacientes mexicanos.

BACKGROUND: American College of Surgeons (ACS) developed the ACS NSQIP surgical risk calculator that predicts the results of elective and emergency surgical procedures. This tool has been useful improving the morbidity and mortality in hospitals in the United States and Canada. OBJECTIVE: To evaluate the usefulness of the ACS NSQIP risk calculator for predicting postoperative complications in Mexican population. METHOD: Prospective, observational, analytical study. Patients undergoing abdominal surgery were recorded, 21 preoperative variables were captured and entered into the calculator. They were followed up to 30 days postoperatively, identifying 14 types of postoperative complications. RESULTS: 109 patients were registered. A comparison was made between the calculated and observed complications, obtaining a good correlation in the complications of cardiac arrest, surgical site infection, reoperation, sepsis and mortality (p < 0.05). CONCLUSIONS: ACS NSQIP risk calculator is useful in the Mexican population, since the score obtained predicts most postoperative complications including mortality. The use of this tool offers an opportunity to improve decision-making in the care of the surgical patient.

ANTECEDENTES: El American College of Surgeons (ACS) desarrolló la calculadora de riesgo quirúrgico ACS NSQIP que predice los resultados de las cirugías electivas y de urgencia. Dicha herramienta ha sido útil para mejorar las cifras de morbilidad y mortalidad en hospitales de los Estados Unidos y Canadá. OBJETIVO: Evaluar la utilidad de la calculadora de riesgo ACS NSQIP para predecir complicaciones posquirúrgicas en pacientes mexicanos. MÉTODO: Estudio prospectivo, observacional y analítico. Se registraron los pacientes sometidos a cirugía abdominal, se capturaron 21 variables preoperatorias y se ingresaron en la calculadora. Se vigilaron hasta cumplir 30 días de posoperatorio y se identificaron 14 tipos de complicaciones posoperatorias. RESULTADOS: Se registraron 109 pacientes y se hizo una comparación entre las probabilidades de complicaciones calculadas y observadas, obteniendo una buena correlación en las complicaciones de paro cardiaco, infección de sitio quirúrgico, reintervención quirúrgica, sepsis y mortalidad (p < 0.05). CONCLUSIONES: La calculadora de riesgo ACS NSQIP es útil en la población mexicana, ya que el puntaje obtenido predice la mayoría de las complicaciones posoperatorias, incluida la mortalidad. El uso de esta herramienta ofrece una oportunidad para mejorar la toma de decisiones en la atención del paciente quirúrgico.

A high-throughput drug screen reveals means to differentiate triple-negative breast cancer.

Plasticity delineates cancer subtypes with more or less favourable outcomes. In breast cancer, the subtype triple-negative lacks expression of major differentiation markers, e.g., estrogen receptor α (ERα), and its high cellular plasticity results in greater aggressiveness and poorer prognosis than other subtypes. Whether plasticity itself represents a potential vulnerability of cancer cells is not clear. However, we show here that cancer cell plasticity can be exploited to differentiate triple-negative breast cancer (TNBC). Using a high-throughput imaging-based reporter drug screen with 9 501 compounds, we have identified three polo-like kinase 1 (PLK1) inhibitors as major inducers of ERα protein expression and downstream activity in TNBC cells. PLK1 inhibition upregulates a cell differentiation program characterized by increased DNA damage, mitotic arrest, and ultimately cell death. Furthermore, cells surviving PLK1 inhibition have decreased tumorigenic potential, and targeting PLK1 in already established tumours reduces tumour growth both in cell line- and patient-derived xenograft models. In addition, the upregulation of genes upon PLK1 inhibition correlates with their expression in normal breast tissue and with better overall survival in breast cancer patients. Our results indicate that differentiation therapy based on PLK1 inhibition is a potential alternative strategy to treat TNBC.

Evaluation of C-reactive protein, procalcitonin and the PCR/PCT index as indicators of mortality in abdominal sepsis.

BACKGROUND: Biomarker combinations can improve timely diagnosis and survival. OBJECTIVE: To determine the usefulness of serum procalcitonin concentration (PCT), C-reactive protein (PCR) and the PCR / PCT index as predictors of mortality. METHOD: Retrospective study of patients diagnosed with abdominal sepsis during the period from April 2017 to February 2018. RESULTS: We included 182 cases. In the survivors, the mean PCR was 170 and procalcitonin (PCT) 10.5. In the deceased, the mean of C-reactive protein (CRP) was 328 and that of PCT was 17.6. When applying the student's t-test for independent samples, it was found that these differences were significant for PCR (p = 0.001); however, for PCT it was not significant (p = 0.460). Afterwards, the PCR/PCT index was studied, as a predictor of mortality, in the deceased cases a PCR/PCT score of 7534 (standard deviation [SD]: 19,303) and for survivors of 538 (SD:805) (p = 0.001) was obtained. CONCLUSION: CRP is associated with mortality, serum PCT does not correlate with mortality. The PCR/PCT index seems to be a better indicator to predict mortality in patients with abdominal sepsis due to secondary peritonitis.

ANTECEDENTES: Las combinaciones de biomarcadores pueden mejorar el diagnóstico oportuno y la supervivencia. OBJETIVO: Determinar la utilidad de la concentración sérica de procalcitonina (PCT), la proteína C reactiva (PCR) y el índice PCR/PCT como predictores de mortalidad. MÉTODO: Estudio retrospectivo de pacientes con diagnóstico de sepsis abdominal durante el periodo de abril de 2017 a febrero de 2018. RESULTADOS: Se incluyeron 182 casos. En los sobrevivientes, la media de los valores de PCR fue de 170 y la de PCT fue de 10.5. En los fallecidos, la media de los valores de PCR fue de 328 y la de PCT fue de 17.6. Al aplicar el estadístico t de Student para muestras independientes se obtuvo que estas diferencias resultaron significativas para la PCR (p = 0.001), pero no para la PCT (p = 0.460). Posteriormente se estudió el índice PCR/PCT como predictor de mortalidad: en los fallecidos se obtuvo un valor de 7534 (desviación estándar [DE]:± 19,303) y en los sobrevivientes de 538 (DE± 805) (p = 0.001). CONCLUSIÓN: La PCR se asocia con la mortalidad, mientras que la PCT no guarda relación con la mortalidad. El índice PCR/PCT parece ser un mejor indicador para predecir la mortalidad en los pacientes con sepsis abdominal por peritonitis secundaria.

Comparison between the body mass index and the Controlling Nutritional status to determine the severity in patients with abdominal sepsis.

BACKGROUND: The systemic response of the organism, in defense against the aggressor agent, generates acute catabolic response, which leads to deterioration of the nutritional status. OBJECTIVE: Compare the usefulness of the body mass index (BMI) and the CONUT scale to determine the severity in abdominal sepsis. METHODS: Retrospective, descriptive, cross-sectional study in patients with diagnosis of abdominal sepsis, from April 2016 to February 2017. RESULTS: We included 153 cases (61 female and 92 male); mean age of 47.44 years, the main organ causing abdominal sepsis was the appendix 43%. Mortality of 15%. An average BMI of 27.31, CONUT score of 5.5, was obtained. The findings, subjected to the Mann-Whitney u test, showed statistical significance when evaluating BMI against SOFA (p = 0.025); no significance was found when evaluating the BMI against APACHE II (p = 0.322), nor against mortality (p = 0.646). Regarding CONUT, significance was found when comparing against APACHE II, SOFA and mortality (p = 0.002, p = 0.001 and p = 0.007, respectively). CONCLUSIONS: The level of malnutrition determined by CONUT is related to the severity determined by APACHE II, SOFA and mortality. BMI is not related to severity by APACHE II or mortality; although it does seem to relate to the severity evaluated by the SOFA scale.

INTRODUCCIÓN: La respuesta sistémica del organismo, en defensa ante el agente agresor, genera una respuesta catabólica aguda que conduce a deterioro del estado nutricional. OBJETIVO: Comparar la utilidad del índice de masa corporal (IMC) y del índice de Control Nutricional (CONUT) para determinar la gravedad en pacientes con sepsis abdominal. MÉTODO: Estudio retrospectivo, descriptivo, transversal, en pacientes con diagnóstico de sepsis abdominal, de abril de 2016 a febrero de 2017. RESULTADOS: Se incluyeron 153 casos (61 mujeres y 92 hombres). El principal órgano causante de sepsis abdominal fue el apéndice (43%). La mortalidad fue del 15%. El IMC promedio fue de 27.31. El CONUT promedio fue de 5.5, Los hallazgos, sometidos a la prueba U de Mann-Whitney, mostraron al evaluar el IMC contra la escala SOFA (Sequential Organ Failure Assessment Score) una p = 0.025; no se encontró significancia al evaluar el IMC contra APACHE II (Acute Physiology and Chronic Health Evaluation) (p = 0.322) ni contra la mortalidad (p = 0.646). En cuanto a CONUT, se encontró significancia al compararla contra APACHE II, SOFA y la mortalidad (p = 0.002, p = 0.001 y p = 0.007, respectivamente). CONCLUSIÓN: El grado de malnutrición determinado por CONUT se relaciona con la gravedad determinada mediante APACHE II y SOFA, y con la mortalidad. El IMC no se relaciona con la gravedad por APACHE II ni con la mortalidad, aunque sí parece relacionarse con la gravedad evaluada mediante SOFA.

Is albumin a predictor of severity and mortality in patients with abdominal sepsis?

BACKGROUND: Patients with a lower level of albumin have a more severe infection, the level of said biomarker is a strong predictor of mortality. OBJECTIVE: To determine the usefulness of the serum albumin level as a predictor of severity and mortality. METHODS: Retrospective, descriptive, cross-sectional study of patients diagnosed with abdominal sepsis. During the period from April 2016-February 2017. The severity was determined by Acute Physiology and Chronic Health Evaluation (APACHE) II, Sequential Organ Failure Assessment (SOFA), Mannheim and mortality. The sample was divided into those with albumin > 2.9 mg/dl and < 2.8 mg/dl. RESULTS: We included 155 cases, 62 female and 93 male; the main organ causing abdominal sepsis was the appendix 42%. The average albumin for the sample was 3.2 mg/dl (DE ± 0.9). The findings, subjected to statistical verification by means of the Mann-Whitney test, showed statistical significance among the cases with albumin < 2.8 mg/dl with those have ranged Mannheim > 26 points (p = 0.001), APACHE > 15 (p = 0.015) and SOFA > 6 (p = 0.001), No statistical significance was obtained between albumin level < 2.8, and mortality (p = 0.052). CONCLUSION: Albumin can be considered as a predictor of severity, although not as a predictor of mortality.

INTRODUCCIÓN: Los pacientes con unos valores más bajos de albúmina presentan una infección más grave; el nivel de dicho biomarcador es un fuerte predictor de la mortalidad. OBJETIVO: Determinar la utilidad de la concentración sérica de albúmina como predictor de gravedad y mortalidad. MÉTODO: Estudio retrospectivo, descriptivo, transversal, de pacientes con diagnóstico de sepsis abdominal, atendidos durante el periodo de abril de 2016 a febrero de 2017. Se determinaron la gravedad mediante APACHE II, SOFA y Mannheim, y la mortalidad. Se dividió la muestra en pacientes con albúmina > 2.9 y < 2.8 mg/dl. RESULTADOS: Se incluyeron 155 casos (62 mujeres y 93 hombres); el principal órgano causante de sepsis abdominal fue el apéndice (42%). El valor medio de la albúmina para la muestra se situó en 3.2 mg/dl (desviación estándar: ± 0.9). Los hallazgos, sometidos a verificación estadística mediante la prueba U de Mann-Whitney, mostraron una relación con significancia estadística entre los casos con albúmina < 2.8 mg/dl y los casos con puntaje de Mannheim > 26 puntos (p = 0.001), APACHE > 15 (p = 0.015) y SOFA > 6 (p = 0.001). No se obtuvo significancia estadística entre el valor de la albúmina < 2.8 y la mortalidad (p = 0.052). CONCLUSIÓN: La albúmina puede ser considerada como un predictor de gravedad, pero no de mortalidad.

A high-throughput screening for inhibitors of riboflavin synthase identifies novel antimicrobial compounds to treat brucellosis.

Brucella spp. are pathogenic intracellular Gram-negative bacteria adapted to life within cells of several mammals, including humans. These bacteria are the causative agent of brucellosis, one of the zoonotic infections with the highest incidence in the world and for which a human vaccine is still unavailable. Current therapeutic treatments against brucellosis are based on the combination of two or more antibiotics for prolonged periods, which may lead to antibiotic resistance in the population. Riboflavin (vitamin B2) is biosynthesized by microorganisms and plants but mammals, including humans, must obtain it from dietary sources. Owing to the absence of the riboflavin biosynthetic enzymes in animals, this pathway is nowadays regarded as a rich resource of targets for the development of new antimicrobial agents. In this work, we describe a high-throughput screening approach to identify inhibitors of the enzymatic activity of riboflavin synthase, the last enzyme in this pathway. We also provide evidence for their subsequent validation as potential drug candidates in an in vitro brucellosis infection model. From an initial set of 44 000 highly diverse low molecular weight compounds with drug-like properties, we were able to identify ten molecules with 50% inhibitory concentrations in the low micromolar range. Further Brucella culture and intramacrophagic replication experiments showed that the most effective bactericidal compounds share a 2-Phenylamidazo[2,1-b][1,3]benzothiazole chemical scaffold. Altogether, these findings set up the basis for the subsequent lead optimization process and represent a promising advancement in the pursuit of novel and effective antimicrobial compounds against brucellosis.

Influencia de la inmunonutrición sobre el estado general, nutricional y estancia hospitalaria en pacientes postoperados de sepsis abdominal / Influence of immunonutrition on the general and nutritional state, and on the in-hospital stay, of patients operated secondary to abdominal sepsis

Objetivo: Valorar el efecto del uso de glutamina con alimentación temprana, en pacientes con sepsis abdominal resuelta quirúrgicamente, sobre el estado general de salud (Apache II), el catabolismo proteico (nitrógeno ureico urinario), las complicaciones y la duración de la estancia hospitalaria. Sede: Hospital General de México. Diseño: Ensayo clínico controlado. Analisis estadístico: Prueba t de Student. Método: Se seleccionaron 24 pacientes con sepsis abdominal que requirieron resolución quirúrgica en la Unidad de Urgencias en el Hospital General de México. Luego de 24 a 72 h del evento quirúrgico, se inició la alimentación enteral en dos grupos: el primero con glutamina más alimentación estándar y el segundo con alimentación estándar. El día 1 y en el 3ro del postoperatorio se midieron niveles de laboratorio y nitrógeno ureico urinario, para determinar la clasificación Apache II, las complicaciones infecciosas y las no infecciosas, el balance nitrogenado y la duración de la estancia hospitalaria. Resultados: No se encontró diferencia significativa en ninguno de los parámetros medidos entre el grupo de glutamina y el control; sin embargo, sí lo hubo en el valor de Apache II en el grupo de glutamina comparando intragrupo entre el primer día y el tercero (P < 0.05). Conclusiones: La alimentación enteral temprana suplementada con glutamina no presenta diferencia temprana en cuanto a la respuesta metabólica al trauma, estado nutricional ni utilización proteica.

Objective: To assess the effect of glutamine, together with early enteral nutrition in patients with surgically resolved abdominal sepsis, on the general health status (Apache II), protein catabolism (urinary urea nitrogen test), complications, and duration of in-hospital stay. Setting: General Hospital of Mexico. Design: Controlled clinical trial. Statistical analysis: Student's t test. Method: We chose 24 patients with abdominal sepsis from the Emergency Ward in the General Hospital of Mexico, who required surgical resolution. After 24 to 72 h of the surgery, enteral nutrition was started in two groups: the first with glutamine plus standard nutrition and the second with standard nutrition. On days 1 and 3 of the postoperative period, laboratory tests were performed and urinary urea nitrogen was measured to determine Apache II classification, infectious and non-infectious complications, the nitrogen balance, and duration of in-hospital stay. Results: No statistically significant difference was found in any of the measured parameters between the two groups (glutamine and control); however, a significant difference was found in the value of the Apache II in the glutamine group when compared intragroup on the first and third days (p < 0.05). Conclusions: Early enteral nutrition supplemented with glutamine does not reveal an early difference in terms of the metabolic response to trauma, nutritional state, or protein utilization.