Sex differences orchestrated by androgens at single-cell resolution.

Sex differences in mammalian complex traits are prevalent and are intimately associated with androgens1-7. However, a molecular and cellular profile of sex differences and their modulation by androgens is still lacking. Here we constructed a high-dimensional single-cell transcriptomic atlas comprising over 2.3 million cells from 17 tissues in Mus musculus and explored the effects of sex and androgens on the molecular programs and cellular populations. In particular, we found that sex-biased immune gene expression and immune cell populations, such as group 2 innate lymphoid cells, were modulated by androgens. Integration with the UK Biobank dataset revealed potential cellular targets and risk gene enrichment in antigen presentation for sex-biased diseases. This study lays the groundwork for understanding the sex differences orchestrated by androgens and provides important evidence for targeting the androgen pathway as a broad therapeutic strategy for sex-biased diseases.

Ferroptosis: an iron-dependent cell death form linking metabolism, diseases, immune cell and targeted therapy / Ferroptosis: una forma de muerte celular dependiente de hierro que vincula el metabolismo, las enfermedades, las células inmunitarias y la terapia dirigida

Compared with the traditional forms of cell death—apoptosis, necrosis and autophagy, ferroptosis is a novel form of iron-dependent programmed cell death forms which is different from the above traditional forms of cell death. Brent R Stockwell, a Professor of Columbia University, firstly proposed that this from of cell death was named ferroptosis in 2012. The main characteristics of ferroptosis is increasing iron loading and driving a lot of lipid peroxide generated and ultimately lead to cell death. In this paper, the mechanism of ferroptosis, relationship between ferroptosis and common diseases and immune state of body are reviewed, and the inhibitors and inducers related to ferroptosis that have been found are summarized to provide medicine exploration targeted of ferroptosis and reference for the research in the future.

Isolated cancer stem cells from human liver cancer: morphological and functional characteristics in primary culture

BackgroundPrimary liver cancer cells (PLCs) could more directly simulate the human tumor microenvironment. Compared with liver cancer cell lines, PLCs could reflect the human situation. As in previous studies, tumor stem cells were a small number of cancer cells in the microenvironment and considered to be one of the origins of liver cancer. This study aimed to screen stem cells in PLCs, analyze their biological characteristics, propose the possibility that liver cancer originated from stem cells.MethodsLiver cancer tissues of 17 patients were taken from the Affiliated Hospital of Guangdong Medical College, and PLCs were isolated by tissue slice method. The proliferation, tumor formation in nude mice, stem protein expression of PLCs were observed. C-kit+ liver cancer cells were screened and their biological characteristics were analyzed.ResultsPLCs could be stably passaged. Transmission electron microscopy indicated that the nucleus was irregular, there were many mitochondria, and the endoplasmic reticulum was irregularly distributed. PLCs could express E-Cadherin, Oct-4, β-Catenin, Sox2, CD326, C-kit, GPC3, Nanog. The proliferation curve of PLCs and Hep3B cells were similar, and they all could form tumors in nude mice. Flow-sorted C-kit+ PLCs, as well as C-kit+ Hep3B cells could highly express Bmi1, Sox2, Oct4, Notch1, Nanog, C-kit, β-Catenin, Smo, Nestin, ABCG2, ABCB1. And they also could clone and form tumors in vivo. But C-kit+ PLCs were more sensitive to chemotherapy drugs than C-kit+ liver cancer cell lines.ConclusionC-kit+ PLCs had the characteristics of tumor stem cells and were more sensitive to chemotherapy drugs.

The peri- and intratumoral immune cell infiltrate and PD-L1 status in invasive squamous cell carcinomas of the penis

IntroductionPenile carcinomas are rare tumors throughout Europe. Therefore, little attention is drawn to this disease. That makes it important to study tumor-associated key metrics and relate these to known data on penile neoplasias.Materials and methodsA cohort of 60 well-defined penile invasive carcinomas with known human papillomavirus (HPV) infection status was investigated. Data on tumor type, grading and staging were recorded. Additionally, data on the peri- and intratumoral immune cell infiltrate in a semiquanititave manner applying an HE stain were assessed.ResultsOur study showed a significant correlation of immune cell infiltrate and pT stage with overall survival. Therefore, in a subset of tumors, PD-L1 staining was applied. For tumor proportion score (TPS), 26 of 30 samples (87%) were scored >0%. For the immune cell score (IC), 28 of 30 samples (93%) were defined as >0% and for CPS, 29 of 30 samples (97%) scored >0. PD-L1 expression was not associated with overall survival.ConclusionPD-L1 is expressed in penile carcinomas, providing a rationale for targeted therapy with checkpoint inhibitors. We were able to show that immune reaction appears to be prognostically relevant. These data enhance the need for further studies on the immune cell infiltrate in penile neoplasias and show that PD-L1 expression is existent in our cohort, which may be a potential target for checkpoint inhibitor therapy.

Selective Cytotoxic effect of Probiotic, Paraprobiotic and Postbiotics of L. casei strains against Colorectal Cancer Cells: Invitro studies

Abstract This study highlights the cytotoxic effect of three L. casei strains on colorectal cell lines in invitro conditions. Different concentrations of live, heat killed (HK) and cell free supernatant (CFS) of three L.casei strains were subjected to CaCo2 and MRC5 cell lines. The viability of the treated and untreated cells was determined after 72 hrs by MTT assay, and IC50 estimated. Apoptosis was evaluated by Annexin V-propidium iodide method using flow cytometry. The live, HK and CFS of the L. casei strains showed cytotoxic effects on colorectal cell lines with significant differences. The cytotoxicity effects of live cells on CaCo2 cells were significantly higher (p˂0.01) than the HK cells. A dose dependent response was observed, as higher concentrations resulted in enhanced cytotoxicity effects. Live L.casei 1296-2cells inhibited 91% of CaCo2 cell growth, with IC50 of less than 108 cfu/ml. MRS medium and concentrations of CFS at above 20% v/v, were cytotoxic to the normal cell lines. Flow cytometry analyses of L. casei 1296-2 indicated that cytotoxicity effects on CaCo2 cells is related to apoptotic induction. Invitro studies indicate that Live and CFS of L. casei 1296-2 might be promising candidate for the control of colorectal cancers

Compostos fenólicos da polpa e sementes de romã ( Púnica granatum, L)atividade antioxidante e protetrora em células MDCK

Os radicais livres, dentre os quais as EROs e as ERNs, são espécies reativas envolvidas no início de reaçõescomo a peroxidação, que no sistema biológico pode dar início ao desenvolvimento de diversos processos patológicos. Para conter o avanço da cadeia oxidativa iniciada pelos radicais livres e minimizar os seus efeitos deletérios ao organismo, algumas substâncias antioxidantes como compostos fenólicos, dentre os quais os ácidos fenólicos, podem ser encontrados em diversas fontes vegetais. Asfrutas são uma boa fonte de compostos antioxidantes, e a romã (Punica granatum, L.) apresenta expressiva quantidade de compostos fenólicos. Os objetivos deste trabalho foram identificar os principais ácidos fenólicos, presentes na fruta (polpa e sementes), através de cromatografi a gasosa, a avaliação da capacidade antioxidante, utilizando dois métodos: o de co-oxidação de substratos (β- caroteno e ácido linoléico) e o da redução do radical 1,1- diphenyl- 2- picryl- hydrazyl (DPPH•). Ainda, foi verifi cado o efeito da fração de ácidos fenólicos livres da polpa da romã (AFLp) sobre a proliferação e viabilidade em cultura de células MDCK. Pôde-se verifi car que a fração AFLp apresentou o maior conteúdo de compostos redutores (1.399,02μg/mL) enquanto que a de ácidos fenólicos ligados a compostos solúveis apresentou o maior conteúdo (770,58 μg/mL) nas sementes(AFESs). Todas as frações de ácidos fenólicos apresentaram resultados signifi cativamente (p<0,05) superiores aos do antioxidante sintético butil-hidroxitolueno, utilizado como padrão. Os principais ácidos fenólicos encontrados foram o salicílico, o protocatequínico e o gálico. As célulastratadas com baixas concentrações da fração AFLp apresentaram aumento da proliferação celular e a viabilidade manteve-se alta.

Metabolic active-high density VERO cell cultures on microcarriers following apoptosis prevention by galactose glutamine feeding

The control of cell death occurring in high density cultures performed in bioreactors is an important factor in production processes. In this work, medium nutrient removal or feeding was used to determine at which extension apoptosis could be, respectively, involved or prevented in VERO cell cultures on microcarriers. Glutamine and galactose present in the VERO cell culture medium was consumed after, respectively, 6 and 12 days of culture. Kinetics studies showed that fresh medium replacement and, to some extent, galactose or glutamine depleted-fresh medium replacement provided a nutritional environment, allowing the VERO cell cultures to attain high densities. Galactose was shown to be a more critical nutrient when cultures reached a high density. In agreement with that, VERO cell cultures supplemented with galactose and/or glutamine were shown to confirm previous findings and, again at high densities, galactose was shown to be a critical nutrient for VERO cell growth. These observations also indicated that in VERO cell cultures, for feeding purposes, the glutamine could be replaced by galactose. The inverse was not true and led, at high densities, to a decrease of cell viability. In the absence of glutamine and galactose, apoptosis was observed in VERO cell cultures by cytofluorometry, Acridine orange staining or light and electron microscopy, reaching high levels when compared to cultures performed with complete medium. VERO cells apoptosis process could be prevented by the galactose and/or glutamine feeding and, at high densities, galactose was more efficient in protecting the cultures. These cultures, prevented from apoptosis, were shown to synthesize high levels of measles virus following infection. Our data show that apoptosis prevention by glutamine/galactose feeding, led to high productive and metabolic active VERO cell cultures, as indicated by the high cell density obtained and the virus multiplication leading to higher virus titers.

Citocinas inflamatorias y factores de reparaci¨®n en los m¨²sculos intercostales de pacientes con EPOC grave / Inflammatory Cytokines and Repair Factors in the Intercostal Muscles of Patients With Severe COPD

Introducci¨®nLas acciones locales de las citocinas en los m¨²sculos de los pacientes con enfermedad pulmonar obstructiva cr¨®nica (EPOC) se hallan sometidas a debate. El objetivo del presente estudio ha sido analizar las relaciones entre su expresi¨®n y la activaci¨®n gen¨¦tica de programas de reparaci¨®n muscular.Pacientes y m¨¦todosSe incluy¨® en el estudio a 25 pacientes con EPOC grave en situaci¨®n estable. Se les realiz¨® una biopsia del m¨²sculo intercostal externo, donde se evaluaron los signos de lesi¨®n muscular (morfometr¨ªa), la infiltraci¨®n de c¨¦lulas inflamatorias (inmunohistoqu¨ªmica) y la expresi¨®n de genes seleccionados (t¨¦cnica de reacci¨®n en cadena de la polimerasa en tiempo real) correspondientes a las propias citocinas ¡ªfactor de necrosis tumoral alfa (TNF-¦Á) y sus receptores 1 y 2 (TNFR1 y TNFR2), e interleucinas-1¦Â, 6 y 10¡ª, un marcador panleucocitario (CD18) y mol¨¦culas clave en las v¨ªas de reparaci¨®n-miog¨¦nesis (Pax7, M-Caderina y Mio-D).ResultadosLa expresi¨®n de TNFR2 se relacion¨® directamente con la funci¨®n muscular inspiratoria (representada por la presi¨®n inspiratoria m¨¢xima sostenible; r=0,496, p<0,05), mientras que la expresi¨®n de CD18 se relacion¨® inversamente con ella (r=−0,462, p<0,05). Por otra parte, la expresi¨®n de los 2 receptores del TNF-¦Á se relacion¨® directamente con la de las mol¨¦culas clave de las v¨ªas de reparaci¨®n analizadas (TNFR1 con Pax7, r=0,650, y M-Caderina, r=0,678, ambas con p<0,001; TNFR2 con Pax7, r=0,395, M-Caderina, r=0,409, y Mio-D, r=0,418, con p<0,05 en todas).ConclusionesLa expresi¨®n de los receptores del TNF-¦Á guarda una estrecha relaci¨®n tanto con la activaci¨®n de los programas de miog¨¦nesis como con la propia funci¨®n muscular inspiratoria. Este hecho refuerza nuestra hip¨®tesis de que algunas citocinas locales participan en la reparaci¨®n de los m¨²sculos respiratorios en los pacientes con EPOC(AU)

ObjectiveThere is disagreement regarding the local action of cytokines in the respiratory muscles of patients with chronic obstructive pulmonary disease (COPD). The objective of this study was to analyze the relationships between cytokine expression and genetic activation of the mechanisms of muscle repair.Patients and methodsTwenty-five patients with severe COPD and in stable condition were enrolled in the study. We performed a biopsy of the external intercostal muscle of the patients and analyzed the specimen for signs of muscle lesion (morphometry), infiltration of inflammatory cells (immunohistochemistry), and expression of selected genes (real-time polymerase chain reaction technique) corresponding to the cytokines (tumor necrosis factor ¦Á [TNF-¦Á] and its type 1 and 2 receptors [TNFR1 and TNFR2], and interleukin [IL] 1¦Â, IL-6, and IL-10), a pan-leukocyte marker (CD18), and key molecules in the repair-myogenesis pathways (Pax7, M-cadherin, and MyoD).ResultsExpression of TNFR2 is directly related to inspiratory muscle function (represented by maximum sustainable inspiratory pressure; r=0.496; P<.05), whereas expression of CD18 is inversely related (r=0.462; P<.05). Moreover, expression of the 2 TNF-¦Á receptors was directly related to that of the key molecules of the repair pathways analyzed (TNFR1 to Pax7 [r=0.650; P<.001] and M-cadherin [r=0.678; P<.001]; TNFR2 to Pax7 [r=0.395; P<.05], M-cadherin [r=0.409; P<.05], and MyoD [r=0.418; P<.05]).ConclusionsExpression of TNF-¦Á receptors bears a close relationship both to activation of the myogenesis programs and to inspiratory muscle function. This reinforces our hypothesis that some local cytokines take part in the repair of respiratory muscles in patients with COPD(AU)

Entecavir / Entecavir

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Losac, a factor X activator from Lonomia obliqua bristle extract Its role in the pathophysiological mechanisms and cell survival

Contact with the bristles of the caterpillar Lonomia obliqua can cause serious hemorrhage. Previously it was reported that a procoagulant protein (Lopap) in the bristle extract of L. obliqua increases cell longevity by inhibiting apoptosis. In this work, we purified from bristle extract a factor X activator that stimulates proliferation of endothelial cells. This protein, named Losac, was purified by ion exchange chromatography, followed by gel filtration chromatography and reverse-phase HPLC. Losac is a 45-kDa protein that activates factor X in a concentration-dependent manner and does not depend on calcium ions. In cultures of HUVECs, Losac increased cell proliferation and inhibited the apoptosis induced by starvation. HUVECs incubated with Losac (0.58 ìM for 1 h) increased release of nitric oxide and tissue-plasminogen activator, which both may mediate anti-apoptosis. Losac also increased slightly the decay-accelerating factor (DAF = CD55), which protects cells from complement-mediated lysis. On the other hand, Losac did not alter the release or expression of von Willebrand factor, tissue factor, intercellular adhesion molecule-1, interleukin-8, and prostacyclin. These characteristics indicate that Losac, a protein with procoagulant activity, also functions as a growth stimulator and an inhibitor of cellular death for endothelial cells. Losac may have biotechnological applications, including the reduction of cell death and consequently increased productivity of animal cell cultures, and the use of hemolymph of L. obliqua for this purpose is already being explored. Further study is required to elucidate the mechanism for the inhibition of apoptosis by Losac.

Envejecimiento vs. radicales libres / Free radicals vs. aging

Los radicales libres se han visto implicados en más de 100 enfermedades tanto humanas como animales, desde artritis reumatoide y shock hemorrágico hasta Sida. Actualmente se estudia su importancia en los procesos de envejecimiento celular, ya que cualquier deficiencia en el sistema antioxidante provoca una pérdida de protección de los tejidos frente al ataque de radicales libres. Niveles anormales de enzimas antioxidantes GPx y SOD, y la reducción del estado de los antioxidantes totales, se han visto implicados en varias enfermedades

Las células B en la artritis reumatoide / B cells in rheumatoid arthritis

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Intervilosite crónica masiva da placenta / Intervilositis crónica masiva de placenta / Massive chronic intervillositis of placeta

A intervilosite crónica massiva é uma patologia raras vezes diagnosticada na placenta, com uma incidência inferior a 0.5%. O exame histológico da placenta mostra uma infiltração massiva do espaço intervilositário por células inflamatórias, sem lesões de vilite crónica associadas. Estas alterações vão comprometer a função placentar originando um compromisso fetal que se manifesta sob a forma de atraso de crescimento fetal intra-uterino, parto pré-termo ou morte fetal. Descreve-se um caso clínico, ilustrativo deste tipo de patologia, referente a uma mulher de 31 anos, II gesta, O para, com antecedentes de morte fetal intra-uterina às 13 semanas de gestação em gravidez anterior, tendo registado na actual gravidez um valor elevado de alfa-feto-proteína às 16 semanas de gestação e morte fetal intra-uterina inesperada às 18 semanas. O exame histológico da placenta demonstrou a presença de uma intervilosite crónica massiva. Este caso realça, ainda, o valor do exame anátomo-patológico da placenta em todos os casos de morte fetal in-trauterina, e a sua importância para o diagnóstico da etiopatogenia e prognóstico de futuras gestações (AU)

La intervilositis crónica masiva es una patología de la placenta, raras veces diagnosticada, con una incidencia inferior al 0,5 0/0. El examen histológico de la placenta muestra una inflamación masiva del espacio intervillositario, por células inflamatorias, sin lesiones de vellosidades crónicas asociadas. Estas alteraciones van a comprometer la función placentaria, originando un compromiso fetal que se manifiesta en una forma de retraso del crecimiento intrauterino, parto pretérmino o muerte fetal. Se describe un caso clínico ilustrativo de este tipo de patología, referente a una mujer de 31 años, secundigestante y primípara, con antecedentes de muerte fetal intrauterina a las 13 semanas de gestación en el embarazo anterior. En este embarazo se registra un valor elevado de la alfa-fetoproteína a las 16 semanas y muerte inesperada a las 18 semanas de embarazo. El examen histológico de la placenta demostró una intervillositis crónica masiva. Este caso realza el valor del examen histológico de la placenta en todos casos de muerte fetal intrauterina y su importancia en el diagnóstico de la etiopatogenia y pronóstico de futuras gestaciones (AU)

Massive chronic intervillositis (MCI) is an infrequently recognized placental lesion which is reported in less than 0.5 0/0 of cases. MCI is characterized by prominent inflammatory infiltrate in the intervillous space in the absence of significant chronic villitis. MCI has been associated with poor pregnancy outcome, including intrauterine growth restriction, preterm delivery and intrauterine fetal death. We report such a case of a 31-year-old woman, gravida 2, para 0, with a past history of one intrauterine fetal death at 13 weeks pregnancy, who became pregnant, and an elevated maternal serum alpha-fetoprotein was noted at 16 weeks gestation. The intrauterine fetal death was diagnosed at 18 weeks gestation. Histological examination of the placenta demonstrated the presence of MCI. This case also underlines the importance of the routine histopathological examination of the placenta in all cases of intrauterine fetal death, in view of its importance in aetiological diagnostic and prognostic information for future pregnancy (AU)

Los linfocitos T CD8+ en la respuesta inmune celular: ¿como funcionan?, ¿como se evaluan? / Lyphocytes T CC8+ in immune cell reaction

Los conocimientos generados en el campo de la función y la actividad de los linfocitos T CD8+, al igual que en la caracterización de las moléculas del Complejo Mayor de Histocompatibilidad han sido parte fundamental para determinar los mecanismos inmune celulares que actúan en el control de procesos infecciosos y tumorales. Los linfocitos T CD8+, tienen la capacidad de reconocer antígenos presentados por las moléculas de clase I en la superficie de las células. Una vez el antígeno es reconocido por los linfocitos, se inicia una serie de vías de activación en el linfocito que llevan finalmente a la eliminación de la célula blanco, mediante mecanismos como citotoxicidad directa o la inducción de muerte celular programada. En este artículo se revisan los principales aspectos de la respuesta celular mediada por los linfocitos T CD8+ que permiten su identificación utilizando técnicas de laboratorio como la prueba de liberación de cromio y la determinación de citocinas por ELISPOT

Linfocitos Th1, Th2 y enfermedad / Lymphocytes Th1, Th2 and disease

Al activarse, los linfocitos T CD4+ muestran una producción de citocinas que usualmente sigue tres patrones distintos y estereotipados; estos patrones son producidos por subpoblaciones discretas que se denominan Th1, Th2 y Th0. Las células Th1 secretan principalmente interferón- (IFN-), interleucina-2 (IL-2) y factor de necrosis tumoral (FNT), citocinas que tienen un papel clave en la respuesta inmune mediada por células y el fenómeno de hipersensibilidad de tipo tardío. Las células Th2 producen IL-4, IL-5, IL-6, IL-9, IL-10 e IL-13, las cuales juegan un papel clave en la generación de la respuesta inmune humoral. Existe una regulación negativa recíproca entre las células Th1 y Th2 ya que ciertas citocinas de las primeras (principalmente IFN-) y de las segundas (IL-4, IL-10) tienen un efecto inhibitorio sobre células Th2 y Th1 respectivamente. Las células Th0 producen citocinas tipo Th1 y Th2. Es posible determinar el patrón de citocinas que están siendo producidas en un tejido, lo que permite inferir si ocurre una activación preferencial de células Th1, Th2 o Th0 en ese sitio. La activación preferencial de subpoblaciones de linfocitos CD4+ tiene un papel importante en la patogenia de enfermedades infecciosas y autoinmunes. Los pacientes con lepra lepromatosa tienen una activación preferencial de células Th2, lo cual conduce a una enfermedad diseminada con ausencia de respuesta inmune celular. La activación preferencial de células Th2 puede ser de importancia en la patogenia de enfermedades autoinmunes generalizadas, en tanto que las células Th1 tienen importancia en padecimientos autoinmunes órgano-específicos. Resulta factible el manipular in vivo la activación preferencial de linfocitos Th1 y Th2, lo cual puede tener implicaciones terapéuticas importantes