Optimization of organotypic cultures of mouse spleen for staining and functional assays

By preserving cell viability and three-dimensional localization, organotypic culture stands out among the newest frontiers of cell culture. It has been successfully employed for the study of diseases among which neoplasias, where tumoral cells take advantage of the surrounding stroma to promote their own proliferation and survival. Organotypic culture acquires major importance in the context of the immune system, whose cells cross-talk in a complex and dynamic fashion to elicit productive responses. However, organotypic culture has been as yet poorly developed for and applied to primary and secondary lymphoid organs. Here we describe in detail the development of a protocol suitable for the efficient cutting of mouse spleen, which overcomes technical difficulties related to the peculiar organ texture, and for optimized organotypic culture of spleen slices. Moreover, we used microscopy, immunofluorescence, flow cytometry and qRT-PCR to demonstrate that the majority of cells residing in spleen slices remain alive and maintain their original location in the organ architecture for several days after cutting. The development of this protocol represents a significant technical improvement in the study of the lymphoid microenvironment in both physiological and pathological conditions involving the immune system

Mistrial or Misdiagnosis: The Importance of Autopsy and Histopathological Examination in Cases of Sudden Infant Bronchiolitis-Related Death

Pediatrics, among all the branches of medicine, is a sector not particularly affected by a high number of claims. Nevertheless, the economic value of the compensation is significantly high, for example, in cases of children who suffered multiple disabilities following perinatal lesions with a long life expectancy. In Italy, most of the claims for compensation concern surgical pathologies and infections. Among these latter, the dominant role is taken by respiratory tract infections. In this context, the purpose of this manuscript is to present a case series of infant deaths in different emergency-related facilities (ambulances, emergency rooms) denounced by relatives. Following these complaints, the autopsy was performed, and subsequent histological examinations revealed the presence of typical and pathognomonic histological findings of acute viral bronchiolitis, whose morphological appearance is poorly reported in the literature. The analysis of these cases made it possible to highlight the following conclusions: the main problems in diagnosing sudden death causes, especially in childhood, are the rapidity of death and the scarce correlation between the preexistent diseases and of the cause of death itself. For all these reasons, the autopsy, either clinical or medicolegal, is mandatory in cases of sudden unexpected infant death to manage claim requests because only the histological examinations performed on samples collected during the autopsy can reveal the real cause of death

Phase Ib study of poly-epitope peptide vaccination to thymidylate synthase (TSPP) and GOLFIG chemo-immunotherapy for treatment of metastatic colorectal cancer patients

ABSTRACT: Thymidylate synthase (TS) is a tumor-associated enzyme critical for DNA replication and main 5′-fluorouracil (5′-FU) target. TSPP/VAC1 is a multi-arm trial phase-Ib trial program aimed to investigate the toxicity and biomodulatory activity of a poly-epitope-peptide vaccine to TS (TSPP) in cancer patients (pts). Here, we present the results of the TSPP/VAC1/arm C trial aimed to evaluate TSPP in combination with chemo-immunotherapy in pretreated metastatic colo-rectal cancer (mCRC) pts. Twenty-nine pts, 14 males and 15 females, received poly-chemotherapy with gemcitabine [GEM; 1,000 mg/sqm, day-1], oxaliplatin [OX; 80 mg/sqm, day-2], levofolinate [100 mg/sqm, days 1–2], bolus/infusional 5′-FU [400 mg/800 mg/sqm, days 1–2], sargramostim [50 μg, days 3–7/q30], and interleukin-2 [sc. 0.5 MIU twice a day, days 8–14/18–30] [GOLFIG-regimen]. Seventeen pts received sc. TSPP injections at escalating dosage [3 pts, 100 µg (DL-1); 3 pts, 200 µg (DL-2) and 11pts, 300 µg (DL-3)] one week after each chemotherapy cycle (concomitant module), while 10 out 12 pts received TSPP (300 µg) after 12 GOLFIG courses [dose level (DL)-0] (sequential module). TSPP MTD was not achieved. Adverse events consisted in swelling/erythema at injection sites (17 cases), G1–2 haematological (16 cases) and gastro-enteric events (12), fever, rhinitis, conjunctivitis, and poly-arthralgia and rise in auto-antibodies [ANA, ENA, c-ANCA, p-ANCA in the DL1–3 pts]. Both treatment-modules showed immunomodulating and antitumor activity (disease-control-rate, DL1–3 and DL0 were 70.6% and 83.3%, respectively) with a better survival recorded in the second group [median OS DL1–3 vs. DL0 = 8 vs. 16 mo, p = 0.049]. The promising long-term survival produced by the sequential treatment module deserves further phase II evaluation

Treponema pallidum (syphilis) antigen TpF1 induces angiogenesis through the activation of the IL-8 pathway (vol 6, 2018)

This corrects the article DOI: 10.1038/srep18785

Optimización de métodos para el estudio del riesgo asociado a la contaminación fecal del agua y alimentos: cuantificación de patógenos y del riesgo de infecciones virales y análisis metagenómico de virus asociados a hepatitis

El agua es esencial para el sostenimiento de la vida en la tierra, por lo tanto, la calidad debe ser controlada para garantizar su uso de forma segura. Los objetivos planteados en esta tesis están orientados a la optimización de métodos para el estudio del riesgo asociado a la contaminación fecal del agua y alimentos. Este trabajo se dividen en tres secciones: La primera parte busca mejorar las herramientas para cuantificar de una manera más precisa los patógenos y el riesgo microbiológico que tiene el consumo de agua contaminada. En la segunda parte se estima el riesgo microbiológico de la utilización de agua regenerada para el riego de vegetales. En la última parte estudiamos pacientes con hepatitis aguda de etiología desconocida para la valoración de virus emergentes o virus nuevos de posible transmisión fecal-oral como posibles causantes de hepatitis. Los objetivos desarrollados en la presente tesis doctoral han dado lugar a una serie de resultados publicados o en proceso de publicación, las principales conclusiones de estos trabajos fueron: (a) El método SMF pudo ser utilizado eficientemente para concentrar simultáneamente los virus, bacterias y protozoos y se desarrolló un método para estimar la concentración real usando la cuantificaciones obtenidas por q(RT)PCR y la predicción de intervalos al 95% mediante “bootstrap” de las recuperaciones; (b) ninguna de los dos planta evaluadas fueron capaces de superar el umbral de <1.00E-6 DALYs pppy considerado como aceptable por la WHO para irrigar vegetales con agua regenerada y (c) la técnica de secuenciación masiva aplicada en esta tesis permitió detectar una gran variedad de secuencias de especies virales en pacientes con hepatitis y las principales familias identificadas fueron Anelloviridae, Caliciviridae y Astroviridae, todas estas relacionadas con transmisión fecal-oral

use of bone health agents bhas in patients pts with metastatic castration resistant prostate cancer mcrpc treated with radium 223 ra 223 after abiraterone abi an interim review of reassure

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