Molecular testing opportunities on cytology effusion specimens: the pre-analytic effects of various body fluid cytology preparation methods on RNA extraction quality and targeted sequencing.

INTRODUCTION: RNA sequencing (RNAseq) analysis is emerging as a clinical research or diagnostic approach for cytologic samples, but there is need for formal comparison of different sample preparation methods in the cytology laboratory to identify which pre-analytic methods could provide alternatives to formalin-fixed paraffin-embedded (FFPE) sections. MATERIALS AND METHODS: We prepared 13 malignant effusions (metastatic estrogen receptor-positive breast cancer) in the cytology laboratory using 6 routine cytologic methods: FFPE cell block, Carnoy's solution, 95% ethanol (EtOH), air-dried and Diff-Quik, ThinPrep, and SurePath preparations. Measurements of RNA quality, expression of 2 multigene expression signatures, molecular subtype, and 4 common activating mutation sites in each preparation were compared with fresh frozen (FF) cell pellet in RNA preservative using distribution of fragment length and concordance correlation coefficient (CCC). RESULTS: The fraction of RNA fragments measuring 200 bases or more (DV200) were 24% higher from cytospins fixed in Carnoy's solution or 95% EtOH than DV200 from FFPE cell blocks. SurePath samples failed RNAseq quality control. There was high concordance of gene expression measurements with FF samples using cytospins fixed in Carnoy's solution, 95% EtOH, Diff-Quik (CCC = 0.829, 0.812, 0.760, respectively), or ThinPrep (CCC = 0.736), but lower using FFPE cell block (CCC = 0.564). The proportion of mutant transcripts was concordant between FF and any cytologic preparation methods. CONCLUSIONS: Cytospin preparations fixed with Carnoy's or 95% ETOH then Papanicolaou stained produced RNAseq results that were equivalent to FF samples and superior to FFPE cell block sections.

Current Evidence on the Protective Effects of Recombinant Human Erythropoietin and Its Molecular Variants against Pathological Hallmarks of Alzheimer's Disease.

Substantial evidence in the literature demonstrates the pleiotropic effects of the administration of recombinant human erythropoietin (rhEPO) and its molecular variants in different tissues and organs, including the brain. Some of these reports suggest that the chemical properties of this molecule by itself or in combination with other agents (e.g., growth factors) could provide the necessary pharmacological characteristics to be considered a potential protective agent in neurological disorders such as Alzheimer's disease (AD). AD is a degenerative disorder of the brain, characterized by an aberrant accumulation of amyloid ß (Aß) and hyperphosphorylated tau (tau-p) proteins in the extracellular and intracellular space, respectively, leading to inflammation, oxidative stress, excitotoxicity, and other neuronal alterations that compromise cell viability, causing neurodegeneration in the hippocampus and the cerebral cortex. Unfortunately, to date, it lacks an effective therapeutic strategy for its treatment. Therefore, in this review, we analyze the evidence regarding the effects of exogenous EPOs (rhEPO and its molecular variants) in several in vivo and in vitro Aß and tau-p models of AD-type neurodegeneration, to be considered as an alternative protective treatment to this condition. Particularly, we focus on analyzing the differential effect of molecular variants of rhEPO when changes in doses, route of administration, duration of treatment or application times, are evaluated for the improved cellular alterations generated in this disease. This narrative review shows the evidence of the effectiveness of the exogenous EPOs as potential therapeutic molecules, focused on the mechanisms that establish cellular damage and clinical manifestation in the AD.

Instrumento pronóstico de neumonía comunitaria complicada en niños / Prognostic instrument for complicated community pneumonia in children

Introducción: La neumonía comunitaria complicada es una entidad con gran impacto sobre la morbimortalidad pediátrica. Objetivo: Desarrollar un instrumento que permita pronosticar a los niños con alto riesgo de desarrollar neumonía comunitaria complicada. Métodos: Se realizó un estudio cuali-cuantitativo en el Hospital General Comandante Pinares de San Cristóbal, en el período 2013-2016 en dos etapas: una etapa diagnóstica inicial, en la que se efectuó un estudio de cohorte prospectivo observacional que permitió estimar la incidencia de neumonía complicada, describir sus características relevantes e identificar factores de riesgo. En una segunda etapa, se elaboró y validó un instrumento pronóstico de esta entidad. Resultados: La incidencia de neumonía complicada fue del 39,5 por ciento, identificándose seis factores de riesgo: hacinamiento [OR 1,9], fumador pasivo [OR 1,76], no lactancia materna exclusiva hasta el sexto mes [OR 1,85], anemia [OR 4,5], morbilidad infecciosa recurrente [OR 1,69] y uso reciente de antimicrobianos [OR 2,38]. Al aplicar el instrumento pronóstico se obtuvo una sensibilidad del 85,47 y una especificidad del 92,74. Conclusiones: La neumonía complicada es una entidad de presentación frecuente, identificándose seis factores de riesgo. El instrumento desarrollado pronostica la posibilidad de desarrollar neumonía complicada en niños con sensibilidad aceptable y alta especificidad(AU)

Introduction: Community complicated pneumonia is an entity with great impact on morbidity and mortality in children. Objective: To develop an instrument that allows prognosticating children at high risk of developing community complicated pneumonia. Methods: A qualitative-quantitative study was carried out at Comandante Pinares General Hospital, San Cristobal municipality, in the period from 2013 to 2016 in two stages: an initial diagnostic stage, in which it was performed a prospective observational cohort study that allowed to estimate the incidence of complicated pneumonia, to describe its relevant characteristics and identify risk factors. In a second stage, it was developed and validated a prognosis instrument for this entity. Results: The incidence of complicated pneumonia was 39.5 por ciento identifying six risk factors: overcrowding (OR 1.9), passive smoking (OR 1.76), not exclusive breastfeeding until the sixth month (OR 1.85), anemia (OR 4.5), recurrent infectious morbidity (OR 1.69), and recent use of antimicrobials (OR 2.38). When the prognosis instrument was applied, it was obtained a sensitivity of 85.47 and a specificity of 92.74. Conclusions: Complicated pneumonia is an entity of frequent presentation. Six risk factors were identified. The instrument predicts the possibility of developing complicated pneumonia in children with acceptable sensitivity and high specificity(AU)

The Recombinant Human Erythropoietin Administered in Neonatal Rats After Excitotoxic Damage Induces Molecular Changes in the Hippocampus.

In vitro and in vivo experimental evidence has contributed important knowledge regarding the antiapoptotic effect mediated by EPO signaling in the damaged brain, particularly through different models with a hypoxic component. However, little emphasis has been placed on the effectiveness of rhEPO administration against cellular alterations caused by in vivo excitotoxicity or on the molecular mechanism that regulates this effect. In this study, we investigated the effects of a single dose of rhEPO on hippocampal damage induced by subcutaneous application of monosodium glutamate (MSG) on postnatal days 1, 3, 5 and 7 in neonatal rats. We found that a dose of 1000 IU/kg of b.w. administered 24 h after MSG had the greatest protective effect. In addition, we analyzed changes in gene expression, particularly in 3 key molecules involved in EPO-mediated signaling (EPO, EPOR and ßcR). We observed that the expression of EPO and EPOR was differentially modified at both the mRNA and protein levels under the evaluated conditions, while the expression of the ßcR gene was substantially increased. Our data suggest that a low dose of rhEPO is sufficient to induce cellular protection under these experimental conditions and that the molecular changes could be a positive feedback mechanism, mediated by reactive astrocytes in association with in vivo neuroprotective mechanisms.

Metal-Binding Pharmacophore Library Yields the Discovery of a Glyoxalase 1 Inhibitor.

Anxiety and depression are common, highly comorbid psychiatric diseases that account for a large proportion of worldwide medical disability. Glyoxalase 1 (GLO1) has been identified as a possible target for the treatment of anxiety and depression. GLO1 is a Zn2+-dependent enzyme that isomerizes a hemithioacetal, formed from glutathione and methylglyoxal, to a lactic acid thioester. To develop active inhibitors of GLO1, fragment-based drug discovery was used to identify fragments that could serve as core scaffolds for lead development. After screening a focused library of metal-binding pharmacophores, 8-(methylsulfonylamino)quinoline (8-MSQ) was identified as a hit. Through computational modeling and synthetic elaboration, a potent GLO1 inhibitor was developed with a novel sulfonamide core pharmacophore. A lead compound was demonstrated to penetrate the blood-brain barrier, elevate levels of methylglyoxal in the brain, and reduce depression-like behavior in mice. These findings provide the basis for GLO1 inhibitors to treat depression and related psychiatric illnesses.

Interactions Between Epilepsy and Plasticity.

Undoubtedly, one of the most interesting topics in the field of neuroscience is the ability of the central nervous system to respond to different stimuli (normal or pathological) by modifying its structure and function, either transiently or permanently, by generating neural cells and new connections in a process known as neuroplasticity. According to the large amount of evidence reported in the literature, many stimuli, such as environmental pressures, changes in the internal dynamic steady state of the organism and even injuries or illnesses (e.g., epilepsy) may induce neuroplasticity. Epilepsy and neuroplasticity seem to be closely related, as the two processes could positively affect one another. Thus, in this review, we analysed some neuroplastic changes triggered in the hippocampus in response to seizure-induced neuronal damage and how these changes could lead to the establishment of temporal lobe epilepsy, the most common type of focal human epilepsy.

Pseudosindactilia y amputación como principales signos del síndrome de bandas amnióticas / Pseudosyndactyly and amputation as the main features of the amniotic band syndrome

Introducción. Las bandas amnióticas causan un espectro muy amplio de malformaciones fetales, desde labio-paladar hendido e hidrocefalia hasta estrangulación y amputación de extremidades; presenta una prevalencia que va de 1:1,200 a 1:15,000 nacimientos. El siguiente caso clínico se reporta con el objetivo de describir los principales signos del síndrome de bandas amnióticas. Caso clínico. Madre de 21 años, primigesta, con antecedente de vulvovaginitis en el primer trimestre, sin tratamiento; control prenatal irregular sin ultrasonidos obstétricos. El embarazo culminó en cesárea a las 30 semanas por presentar oligohidramnios severo. Se obtuvo producto femenino de 1,200 g y Apgar 7-8. A la exploración física se encontró pseudosindactilia con zona de anillo uniendo a las falanges por el vértice, pie equino varo izquierdo y pierna derecha amputada desde la región tibial media terminando en punta, con escara en porción distal. Además, en estudio radiológico se observaron las falanges del segundo, tercer y cuarto dedos unidas al centro en vértice y peroné hipoplásico. Conclusiones. Los anillos de constricción, la amputación de una o más extremidades y la pseudosindactilia son signos importantes y consistentes que apoyan el diagnóstico del síndrome de bandas amnióticas, con un mal pronóstico para la función aunque bueno para la vida.

Background. Amniotic band syndrome causes a wide spectrum of congenital defects such as cleft lip, hydrocephalus, and growth restriction of limbs, with or without amputation. The condition occurs in every 1:1,200 to 1:15,000 deliveries. The objective of this report is to define the most important features of amniotic band syndrome. Case report. We report the case of a 21 -year-old female with a first pregnancy. No prenatal care or ultrasound studies were carried out, but the patient reported experiencing vaginal discharge during the first trimester. The pregnancy resulted in cesarean section at 30 gestational weeks because of severe oligohydramnios. A 1200-g female was delivered with Apgar 7-8. The newborn presented congenital amputation of the right leg and pseudosyndactyly of the left hand with a ring constriction of the third phalange of the second, third and fourth finger, a left equinovarus foot and amputation of the right leg from the middle third of the tibial region. These findings were confirmed by x-ray. Conclusions. Constriction rings, limb amputation and pseudosyndactyly are important and consistent features with the amniotic band syndrome. Functional prognosis is nonfavorable, but prognosis for life productivity is good.

Condiciones asociadas a la mortalidad neonatal específica en < 1000 g, en el Hospital de Gineco-Obstetricia N° 23, periodo 2005-2006 / Conditions associated with mortality in neonates weighing < 1000 g at the Obstetric-Gynecology Hospital, Unit N° 23, Monterrey, Mexico (2005-2006)

Introducción. La mortalidad en pacientes <1000 g de peso ha disminuido de 90% a 55%, desde 1970 al 2000, pero se han incrementado las secuelas y la estancia intrahospitalaria. Métodos. Para conocer las causas de mortalidad en neonatos de <1000 g, se revisaron certificados y registros hospitalarios y para cuantificar se usaron tasas, medidas de dispersión y proporciones. Resultados. En el periodo de agosto-2005 a julio-2006 nacieron 18, 952 pacientes; 152 (0.8%) eran de <1000 g de peso. De estos pacientes fallecieron 98 (64.4%), 54 masculinos y 44 femeninos (1.2:1). El peso promedio fue de 760 g y la edad gestacional promedio fue de 26 semanas (22 a 35). El Apgar al minuto fue de 4-5 en promedio y 6-7 a los 5 minutos. Las causas de defunción fueron inmadurez en 28/98 casos (28%), hemorragia intracraneana (HIC) 22/98 (22%), sepsis 17/98 (17%), hemorragia pulmonar 9/98 (9%) y falla orgánica múltiple 3/98 (3%). Conclusiones. Se observó 64% de mortalidad en pacientes de <1000 g de peso. Las causas de muerte fueron: inmadurez, HIC y sepsis; la mitad de las muertes ocurrieron en las primeras 48 horas de vida.

Background. Mortality in neonates <1 000 g has decreased since the 1970s until now from 90% to 55%, but neurological sequelae and hospital stay have increased. We undertook this study in order to determine the prevalence and etiology of mortality in neonates <1 000 g in an Ob-Gyn Unit of the IMSS. Methods. Causes of death in neonates weighing <1 000 g were obtained from hospital registries using dispersion, rates and proportions. Results. During a 1-year period, 18 952 neonates were born; 152 (0.8%) weighed <1 000 g and 98(64%) did not survive. There were 54 (55.1%) males and 44 (44.9%) females (1.2:1). The mean weight was 760 g, and mean gestational age was 26 weeks (range: 22-35 weeks). The principal causes of deaths were immaturity in 28/98 cases (28%), intracranial hemorrhage (ICH) in 22/98 (22%), sepsis in 17/98 (17%), pulmonary hemorrhage in 9/98 (9%) and multiple organ failure in 3/98 (3%). Conclusion. Mortality in neonates with very low birth weight (VLBW, <1 000 g) was 64%. Immaturity, ICH and sepsis were the main causes of death. Half of these deaths occurred during the first 48 h of life.

Takotsubo cardiomyopathy: an unusual cardiomyopathy at an unusual age.

We describe an atypical presentation of stress-induced cardiomyopathy - Takotsubo cardiomyopathy - in a 16-month-old boy previously diagnosed with cyclic vomiting and episodic hypertension. He developed features of cardiac failure and his echocardiogram showed left ventricular wall motion abnormality accompanied with elevated cardiac enzymes. Cardiac catheterisation showed no coronary arterial abnormality. Complete spontaneous recovery occurred 2 weeks after admission.

SMc01553 is the sixth acyl carrier protein in Sinorhizobium meliloti 1021.

Acyl carrier proteins (ACPs) are required for the transfer of acyl intermediates during fatty acid and polyketide syntheses. In Sinorhizobium meliloti 1021 there are five known ACPs: AcpP, NodF, AcpXL, the ACP domain in RkpA and SMb20651. The genome sequence of S. meliloti 1021 also reveals the ORF SMc01553, annotated as a putative ACP. smc01553 is part of a 6.6 kb DNA region that is duplicated in the chromosome and in the pSymb plasmid, the result of a recent duplication event. SMc01553 overexpressed in Escherichia coli was labelled in vivo with [(3)H]beta-alanine, a biosynthetic building block of the 4'-phosphopantetheine prosthetic group of ACPs. The purified SMc01553 was modified with 4'-phosphopantetheine in the presence of S. meliloti holo-ACP synthase, and this modification resulted in a major conformational change of the protein structure, since the holo-form runs faster in native PAGE than the apo-form. SMc01553 could not be loaded with a malonyl group by malonyl-CoA-ACP transacylase from S. meliloti. Using RT-PCR we could show the presence of mRNA for SMc01553 and of the duplicated ORF SMb22007 in cultures of S. meliloti. However, a mutant in which the two duplicated regions were deleted did not show any different phenotype with respect to the wild-type in the free-living or symbiotic lifestyle.

SMb20651 is another acyl carrier protein from Sinorhizobium meliloti.

Acyl carrier proteins (ACPs) are small acidic proteins that carry growing acyl chains during fatty acid or polyketide synthesis. In rhizobia, there are four different and well-characterized ACPs: AcpP, NodF, AcpXL and RkpF. The genome sequence of Sinorhizobium meliloti 1021 reveals two additional ORFs that possibly encode additional ACPs. One of these, smb20651, is located on the plasmid pSymB as part of an operon. The genes of the operon encode a putative asparagine synthetase (AsnB), the predicted ACP (SMb20651), a putative long-chain fatty acyl-CoA ligase (SMb20650) and a putative ammonium-dependent NAD+ synthetase (NadE1). When SMb20651 was overexpressed in Escherichia coli, [3H]beta-alanine, a biosynthetic building block of 4'-phosphopantetheine, was incorporated into the protein in vivo. The purified SMb20651 was modified with 4'-phosphopantetheine in the presence of S. meliloti holo-ACP synthase (AcpS). Also, holo-SMb20651 was modified in vitro with a malonyl group by malonyl CoA-ACP transacylase. In E. coli, coexpression of SMb20651 together with other proteins such as AcpS and SMb20650 led to the formation of additional forms of SMb20651. In this bacterium, acylation of SMb20651 with C12 : 0 or C18 : 0 fatty acids was detected, demonstrating that this protein is involved in fatty acid biosynthesis or transfer. Expression of SMb20651 was detected in S. meliloti as holo-SMb20651 and acyl-SMb20651.

Alteraciones de la mucosa bucal en pacientes rehabilitados protésicamente en el Hospital General "Abel Santamaría" / Oral mucous disorders in prosthetic dentistry patients: “Abel Santamaria Cuadrado” University Hospital

Teniendo en cuenta que en Cuba el Programa del Adulto Mayor está insertado dentro de los cuatro programas priorizados del MINSAP y que este grupo poblacional constituye una cifra importante de los pacientes que acuden a consulta, se realiza un estudio descriptivo transversal en los meses comprendidos de enero a marzo del año 2006, el cual tiene como universo a los pacientes portadores de prótesis removibles totales o parciales, mayores de 60 años, que radican en el área de salud atendida por el consultorio 75, ubicado en el Reparto Montequín, perteneciente al policlínico “Hermanos Cruz” y atendido por la consulta de Prótesis del Departamento de Estomatología del Hospital General Docente “Abel Santamaría”. Se selecciona una muestra de pacientes para hacerles un examen bucal; con el objetivo de identificar lesiones en su mucosa bucal que puedan estar relacionadas con el estado funcional de las prótesis y ciertos hábitos de uso por ellos practicados, encontrándose como lesión más frecuente la úlcera traumática, hábitos de higiene inadecuados en gran número de personas entrevistadas, así como pacientes portadores de prótesis confeccionadas desde un prolongado tiempo. Estas lesiones atentan contra el equilibrio biopsicosocial de los individuos y algunas pueden dar origen a estados premalignos, por lo que esta investigación representa un gran beneficio para estos pacientes, pues de diagnosticarse alguna patología serán atendidos por el personal calificado que les devolverá su salud. Los resultados obtenidos se recogen por medio de un cuestionario; se procesan estadísticamente y se muestran en tablas (AU)

Considering “The Cuban Program for the Aged “a descriptive, cross-sectional study was carried out from January to March 2006 taking as sample the patients over 60 years old who had partial or total removable prosthesis. The patients belonged to the health area of “Hermanos Cruz “Doctors Office # 75 located in “Reparto Montequín “. The Dentistry Department at Abel Santamaria University Hospital offers service to these patients. They were examined with the purpose of identifying oral mucous lesions related to the functional conditions of the prosthesis and the practice of certain habits. The most frequent lesion was the traumatic ulcer; besides inadequate hygiene habits and the use of the same prosthesis for long periods of time. These lesions damage the psychosocial equilibrium of the individuals and some of them can provoke pre-malignant conditions, being this study of great help to diagnose the pathologies associated with the use of dentistry prosthesis. Results obtained were collected through questionnaires, statistical analysis was conducted and showed in tables (AU)

[Prenatal diagnosis of fetal cardiac defect]. / Diagnóstico prenatal de cardiopatía fetal.

INTRODUCTION: The prevalence of congenital cardiac defects is 8 per 1000 neonates, and it's different if high or low risk populations are studied. The fetal ultrasonographic increase prenatal detection but varies from 7 to 90%. OBJECTIVES: To know the prevalence of fetal cardiopathy and detection in high risk pregnancies. PATIENTS AND METHODS: A observational study was made in pregnancies women with 16 old week of gestation. RESULTS: We received a total of 3500 high-risk pregnancies and were detected 112 cases with fetal cardiopathy (3.2%). The 30% of them had a risk factor of cardiopathy. The most frequent fetal cardiac defect detected were arrhythmia in 34 fetus, septal defects in 30, valvular defects in 17, hypoplasic or absence of cardiac cavities 16, tronco-conus defects 8, and other 7 included ectopia cordis 3, cardiac tumor 2, abnormal drainage of pulmonary veins 2. The diagnosis increased every year since started study. The prenatal diagnoses suspected in fetal echocardiography were confirmed in 80% of the cases in neonatal period. CONCLUSION: The detection rate of fetal cardiac defect was 3.2% in high-risk pregnancies, four times higher than general population prevalence of congenital heart disease. We found a 30% overall perinatal mortality in fetal cardiac defect. The most frequent fetal cardiac defects found in this screening were arrhythmias and septal ventricular defects in almost 50% of patients.

Determinacion serica de los niveles de ICAM-1s en pacientes con rinitis alergica / Serum levels of ICAM-1s in patient with allergic rhinitis

Se presume que la molécula de adhesión intercelular -1 (ICAM-1, CD54) está involucrada en la patogénesis del asma y se ha detectado una forma soluble de ICAM-1 (ICAM-1 s) en concentraciones elevadas en el suero de pacientes con ciertas condiciones inflamatorias. En este estudio, examinamos los niveles de ICAM-1 soluble en pacientes con rinitis alérgica de diversos grados: leve (8 pacientes), moderada (12 pacientes), severa (15 pacientes), del instituto de alergias, asma e inmunología. Los resultados fueron luego comparados con 10 sujetos saludables, como control negativo, sin diagnóstico de rinitis alérgica ni asma, y con 10 sujetos con asma de la unidad de neumología del Hospital La Samaritana, como control positivo. Todos los pacientes con edades entre 15 y 55 años. Utilizamos la técnica ELISA para medir ICAM-ls. Con anterioridad, todos los pacientes fueron sometidos a una prueba alérgica mediante el método Prick. Nuestro objetivo fue analizar si ICAM-1 s es un marcador de la severidad de la rinitis alérgica, y encontramos que los niveles de ICAM-1 soluble muestran un incremento a medida que la rinitis alérgica progresa. La comparación entre el grupo general de rinitis alérgica y el grupo de control negativo muestra resultados estadísticamente significativos (P < 0.05) y cada uno de los grupos de rinitis alérgica (leve, moderada y severa) en relación con el control negativo mostró que la expresión de ICAM-1 no incrementa y se mantiene baja, (Rinitis Leve p = 0.495, Moderada p = 0.0017, Severa p = 0.0014). Los anteriores resultados confirman nuestra hipótesis

Informe de los resultados de 20847 cultivos de líquido Cefalorraquídeo en el Hospital Escuela 1980-1992 / Report of results of 20847 cultures of cerebrospinal fluid at the Hospital Escuela. 1980-1992

Se revisaron los resultados de 20847 cultivos de muestras de líquido cefalorraquídeo que fueron estudiadas en el laboratorio de Bacteriología del Hospital Escuela. 1932 (9.2 por ciento ) mostraron crecimeinto (positivos), 18884 (90.5 por ciento ) no mostraron ningún crecimiento (negativo). 31 (0.3 por ciento ) resultaron obviamente cpontaminados. Las bacterias más comunmente aisladas en todo periodo fueron, en orden descendente de frecuencia: Staphylococcus sp. coagulasa-negativo (CN), Bacilos Gram Negativo que no fermetan la glucosa (BGN NFG)exceptuendo Pseudomonas aeruginosa, Streptococcus pneumoniae,Pseudomonas aeruginosa, Escherichia coli,Klebsiella pneumonie, Haemophilus influenzae y Staphylococcus aureus. El estudio no debe considerarse un estudio de las causas de meningitis, es únicamente un análisis de los resultados bacteriológicos que sirve de base para proceder a investigar más específicas. Se hacen comentarios y recomendaciones acerca de los hallaszgos encontrados