The 3'-untranslated region directs ribosomal protein-encoding mRNAs to specific cytoplasmic regions

mRNA localization is a conserved post-transcriptional process crucial for a variety of systems. We have analyzed the subcellular distribution of mRNAs encoding human cytosolic and mitochondrial ribosomal proteins. Biochemical fractionation experiments showed that the transcripts for cytosolic ribosomal proteins associate preferentially with the cytoskeleton via actin microfilaments. Transfection in HeLa cells of a GFP reporter construct containing the cytosolic ribosomal protein L4 3'-UTR showed that the 3'-UTR is necessary for the association of the transcript to the cytoskeleton. Using confocal analysis we demonstrate that the chimeric transcript is specifically associated with the perinuclear cytoskeleton. We also show that mRNA for mitochondrial ribosomal protein S12 is asymmetrically distributed in the cytoplasm. In fact, this transcript was localized mainly in the proximity of mitochondria, and the localization was 3'-UTR-dependent. In summary, ribosomal protein mRNAs constitute a new class of localized transcripts that share a common localization mechanism

Non-epithelial ovarian cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up

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Performance Evaluation of Two Stages Parallel Flow Indirect Evaporative Cooling with Aspen Pads in Baghdad Climate Conditions

This study delves into the realm of advanced cooling techniques by examining the performance of a two-stage parallel flow indirect evaporative cooling system enhanced with aspen pads in the challenging climate of Baghdad. The objective was to achieve average air dry bulb temperatures (43 oC) below the ambient wet bulb temperatures (24.95 oC) with an average relative humidity of 23%, aiming for unparalleled cooling efficiency. The research experiment was carried out in the urban environment of Baghdad, characterized by high temperature conditions. The investigation focused on the potential of the two-stage parallel flow setup, combined with the cooling capability of aspen pads, to surpass the limitations imposed by conventional cooling methods. The empirical findings underscored the capacity of the two-stage parallel flow configuration to achieve air-dry bulb temperatures surpassing the ambient wet bulb temperature. Nonetheless, it was evident that an excessive application of water (exceeding 9 lpm) onto the aspen pads within the wet channels yielded adverse repercussions on the cooler's performance metrics, specifically in terms of wet bulb effectiveness and coefficient of performance. A range of air flows spanning from 150 CMH to 350 CMH was analyzed, revealing the influence of heightened air flow rates on the resultant dry bulb temperatures delivered by the cooling system. Notably, the peak wet-bulb effectiveness registered at 1.08, concurrently yielding a coefficient of performance measuring 7.8

Innate type 2 immunity in helminth infection is induced redundantly and acts autonomously following CD11c+ cell depletion

Infection with gastrointestinal helminths generates a dominant type 2 response among both adaptive (Th2) and innate (macrophage, eosinophil, and innate lymphoid) immune cell types. Two additional innate cell types, CD11chigh dendritic cells (DCs) and basophils, have been implicated in the genesis of type 2 immunity. Investigating the type 2 response to intestinal nematode parasites, including Heligmosomoides polygyrus and Nippostrongylus brasiliensis, we first confirmed the requirement for DCs in stimulating Th2 adaptive immunity against these helminths through depletion of CD11chigh cells by administration of diphtheria toxin to CD11c.DOG mice. In contrast, responsiveness was intact in mice depleted of basophils by antibody treatment. Th2 responses can be induced by adoptive transfer of DCs, but not basophils, exposed to soluble excretory-secretory products from these helminths. However, innate type 2 responses arose equally strongly in the presence or absence of CD11chigh cells or basophils; thus, in CD11c.DOG mice, the alternative activation of macrophages, as measured by expression of arginase-1, RELM-α, and Ym-1 (Chi3L3) in the intestine following H. polygyrus infection or in the lung following N. brasiliensis infection, was unaltered by depletion of CD11c-expressing DCs and alveolar macrophages or by antibody-mediated basophil depletion. Similarly, goblet cell-associated RELM-β in lung and intestinal tissues, lung eosinophilia, and expansion of innate lymphoid (“nuocyte”) populations all proceeded irrespective of depletion of CD11chigh cells or basophils. Thus, while CD11chigh DCs initiate helminth-specific adaptive immunity, innate type 2 cells are able to mount an autonomous response to the challenge of parasite infection

Charcot-Leyden Crystals activate the NLRP3 inflammasome and cause IL-1β inflammation [preprint]

Charcot-Leyden crystals (CLCs) are Galectin-10 protein crystals that can form after eosinophils degranule. CLCs can appear and persist in tissues from patients with eosinophilic disorders, such as asthma, allergic reactions, fungal, and helminthic infections. Despite abundant reports of their occurrence in human disease, the inflammatory potential of CLCs has remained unknown. Here we show that CLCs induce IL-1β release upon their uptake by primary human macrophages in vitro, and that they induce inflammation in vivo in mouse models of acute peritonitis and bronchitis. CLC-induced IL-1β was dependent on NLRP3 and caspase-1, and their instillation in inflammasome reporter mice promoted the assembly of ASC complexes and IL-1β secretion in the lungs. Our findings reveal that CLCs are recognized by the NLRP3 inflammasome, which may sustain inflammation that follows eosinophilic inflammatory processes

The Pervasive Effects of ER Stress on a Typical Endocrine Cell: Dedifferentiation, Mesenchymal Shift and Antioxidant Response in the Thyrocyte

none13noThe endoplasmic reticulum stress and the unfolded protein response are triggered following an imbalance between protein load and protein folding. Until recently, two possible outcomes of the unfolded protein response have been considered: life or death. We sought to substantiate a third alternative, dedifferentiation, mesenchymal shift, and activation of the antioxidant response by using typical endocrine cells, i.e. thyroid cells. The thyroid is a unique system both of endoplasmic reticulum stress (a single protein, thyroglobulin represents the majority of proteins synthesized in the endoplasmic reticulum by the thyrocyte) and of polarized epithelium (the single layer of thyrocytes delimiting the follicle). Following endoplasmic reticulum stress, in thyroid cells the folding of thyroglobulin was disrupted. The mRNAs of unfolded protein response were induced or spliced (X-box binding protein-1). Differentiation was inhibited: mRNA levels of thyroid specific genes, and of thyroid transcription factors were dramatically downregulated, at least in part, transcriptionally. The dedifferentiating response was accompanied by an upregulation of mRNAs of antioxidant genes. Moreover, cadherin-1, and the thyroid (and kidney)-specific cadherin-16 mRNAs were downregulated, vimentin, and SNAI1 mRNAs were upregulated. In addition, loss of cortical actin and stress fibers formation were observed. Together, these data indicate that ER stress in thyroid cells induces dedifferentiation, loss of epithelial organization, shift towards a mesenchymal phenotype, and activation of the antioxidant response, highlighting, at the same time, a new and wide strategy to achieve survival following ER stress, and, as a sort of the other side of the coin, a possible new molecular mechanism of decline/loss of function leading to a deficit of thyroid hormones formation.openUlianich L.; Mirra P.; Garbi C.; Cali G.; Conza D.; Treglia A.S.; Miraglia A.; Punzi D.; Miele C.; Raciti G.A.; Beguinot F.; Consiglio E.; Di Jeso B.Ulianich, L.; Mirra, P.; Garbi, C.; Cali, G.; Conza, D.; Treglia, A. S.; Miraglia, A.; Punzi, D.; Miele, C.; Raciti, G. A.; Beguinot, F.; Consiglio, E.; Di Jeso, B

IL-33 alarmin and its active proinflammatory fragments are released in small intestine in Celiac disease

Initially described as Th2 promoter cytokine, more recently, IL-33 has been recognized as an alarmin, mainly in epithelial and endothelial cells. While localized in the nucleus acts as a gene regulator, it can be also released after injury, stress or inflammatory cell death. As proinflammatory signal, IL-33 binds to the surface receptor ST2, which enhances mast cell, Th2, regulatory T cell, andinnate lymphoid cell type 2 functions. Besides these Th2 roles, free IL-33 can activate CD8+ T cells during ongoing Th1 immune responses to potentiate its cytotoxic function. Celiac Disease (CD) is a chronic inflammatory disorder characterized by a predominant Th1 response responsible for multiple pathways of mucosal damage in the proximal small intestine. By immunofluorescence and western blot analysis of duodenal tissues, we found an increased expression of IL-33 in duodenal mucosa of active CD (ACD) patients. Particularly, locally digested IL-33 releases active 18/21kDa fragments which can contribute to expand the proinflammatory signal. Endothelial (CD31+) and mesenchymal, myofibroblast and pericytes, cells from microvascular structures in villi and crypts, showed IL-33 nuclear location; while B cells (CD20+) showed a strong cytoplasmic staining.Both ST2 forms, ST2L and sST2, were also upregulated in duodenal mucosa of CD patients. This was accompanied by increased number of CD8+ST2+ T cells and the expression of T-bet in some ST2+ intraepithelial lymphocytes and lamina propria cells. IL-33 and sST2 mRNA levels correlated with IRF1, an IFN induced factor relevant in responses to viral infections and interferon mediatedproinflammatory responses highly represented in duodenal tissues in ACD. These findings highlight the potential contribution of IL-33 and its fragments to exacerbate the proinflammatory circuit and potentiate the cytotoxic activity of CD8+ T cells in CD pathology.Fil: Perez, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Ruera, Carolina Naymé. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Miculán, Emanuel Gonzalo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Carasi, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Dubois Camacho, Karen. Universidad de Chile. Facultad de Medicina. Institutos de Ciencias Biomedicas.; ChileFil: Garbi, Laura. Provincia de Buenos Aires. Hospital Interzonal General de Agudos Gral. San Martín; ArgentinaFil: Guzman, Luciana. Provincia de Buenos Aires. Ministerio de Salud. Hospital de Niños "Sor María Ludovica" de La Plata; ArgentinaFil: Hermoso, Marcela A.. Universidad de Chile. Facultad de Medicina. Institutos de Ciencias Biomedicas.; ChileFil: Chirdo, Fernando Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; Argentin

Endoscopic dilation in the treatment and profilaxy of esofagien strictures in newborns operated for esophagus atrezia

Endoscopie, Clinica Chirurgie, IMSP Institutul Mamei și Copilului, Chișinău, Republica Moldova, S. C. Donica & Donica, Mangalia, România, Al XIII-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” și al III-lea Congres al Societății de Endoscopie, Chirurgie miniminvazivă și Ultrasonografie ”V.M.Guțu” din Republica MoldovaIntroducere: Restabilirea tranzitului esofagian după plastia chirurgicală esofagiană este subiect recunoscut. Material și metode: Studiul a cuprins 16 copii cu vârsta: două zile de la naștere și șase ani, spitalizați si operați de atrezie de esofag în clinica chirurgie pediatrică IMșiC în perioada 2012-2018. Dilatatoarele folosite: balonașe și bujii Savary-Gilliard. Rezultate: În perioada preoperatorie endoscopic a fost constatată discontinuitatea esofagului cu, sau fără, prezența fistulei esolaringo-traheală. Postoperator s-a remarcat: restrângerea segmentelor individuale ale esofagului și imposibilitatea consecutivă a depașirii stricturii cu endoscopul corespunzător dimensiunilor esofagului; bride mucosale; fire restante; corpi străini. Localizarea și extensia stricturii de la gingii, datorită tehnologiei operațiilor a fost unică la toți pacienții. Stricturile au avut următoarele dimensiuni de apertură: până la 9 mm– 3 copii, 6-7 mm – 8, 4-5 mm - 4 si 2 mm – un copil. La o sesiune pacienții au suportat 3-5 dilatări. Dinamică pozitivă s-a înregistrat la toți copiii. Concluzii: Endoscopia evaluează diferențiat rezultatele plastiei chirurgicale esofagiene, determină tratamentul endoluminal al stricturii anastomozei. Pacienții cu atrezie de esofag operați, necesită dispanserizare în primul an de viața, pentru depistarea posibililor complicații – stenoza anastomozei, incompetență joncțională, refluxul gastro-esofagian. Strictura anastomozei trebuie dilatată sub control vizual, urmată de dilatări de susținere, după necesitate.Introduction: Restoring esophageal transit after esophageal surgery is a recognized issue. Material and Methods: Study includes 16 children aged from two days to six years, operated for esophagus atresia during 2012-2018 years. Used dilators: balloons and Savary-Gillard bougies. Results: Endoscopy revealed discontinuity of the esophagus, the presence or not of eso-laryngo-tracheal fistula. It was noted: the restriction of the individual segments of the esophagus and the consequent impossibility to overpass stricture with the corresponding endoscope; mucous membranes; residual threads; foreign bodies. The location and extension of stinging from the gums due to the operation technology was unique in all patients. The stricture had the following aperture dimensions: up to 9 mm - 3 children, 6-7 mm - 8, 4-5 mm - 4 and 2 mm - one. At one session the patients underwent 3-5 dilations. Positive dynamics was recorded in all children. Conclusions: Endoscopy evaluates the results of surgical esophageal plasty, makes available the endoluminal treatment of anastomosis strictures. Patients with operated esophagus atresia require first-year dispanserization for detection of possible complications - anastomosis stenosis, junctional incompetence, gastro-esophageal reflux. Anastomotic strictures should be dilated under visual control, followed by maintaining dilations

praja2 regulates KSR1 stability and mitogenic signaling

The kinase suppressor of Ras 1 (KSR1) has a fundamental role in mitogenic signaling by scaffolding components of the Ras/MAP kinase pathway. In response to Ras activation, KSR1 assembles a tripartite kinase complex that optimally transfers signals generated at the cell membrane to activate ERK. We describe a novel mechanism of ERK attenuation based on ubiquitin-dependent proteolysis of KSR1. Stimulation of membrane receptors by hormones or growth factors induced KSR1 polyubiquitination, which paralleled a decline of ERK1/2 signaling. We identified praja2 as the E3 ligase that ubiquitylates KSR1. We showed that praja2-dependent regulation of KSR1 is involved in the growth of cancer cells and in the maintenance of undifferentiated pluripotent state in mouse embryonic stem cells. The dynamic interplay between the ubiquitin system and the kinase scaffold of the Ras pathway shapes the activation profile of the mitogenic cascade. By controlling KSR1 levels, praja2 directly affects compartmentalized ERK activities, impacting on physiological events required for cell proliferation and maintenance of embryonic stem cell pluripotency

The impact of chorionicity on pregnancy outcome and neurodevelopment at 2 years old among twins born preterm: the EPIPAGE-2 cohort study

OBJECTIVE To compare the short‐ and mid‐term outcomes of preterm twins by chorionicity of pregnancy. DESIGN Prospective nationwide population‐based EPIPAGE‐2 cohort study. SETTING 546 maternity units in France, between March and December 2011. POPULATION A total of 1700 twin neonates born between 24 and 34 weeks of gestation. METHODS The association of chorionicity with outcomes was analysed using multivariate regression models. MAIN OUTCOME MEASURES First, survival at 2‐year corrected age with or without neurosensory impairment, and second, perinatal, short‐, and mid‐term outcomes (survival at discharge, survival at discharge without severe morbidity) were described and compared by chorionicity. RESULTS In the EPIPAGE 2 cohort, 1700 preterm births were included (850 twin pregnancies). In all, 1220 (71.8%) were from dichorionic (DC) pregnancies and 480 from monochorionic (MC) pregnancies. MC pregnancies had three times more medical terminations than DC pregnancies (1.67 versus 0.51%, P < 0.001), whereas there were three times more stillbirths in MC than in DC pregnancies (10.09 versus 3.78%, P < 0.001). Both twins were alive at birth in 86.6% of DC pregnancies compared with 80.0% among MC pregnancies (P = 0.008). No significant difference according to chorionicity was found regarding neonatal deaths and morbidities. Likewise, for children born earlier than 32 weeks, the 2‐year follow‐up neurodevelopmental results were not significantly different between DC and MC twins. CONCLUSIONS This study confirms that MC pregnancies have a higher risk of adverse outcomes. However, the outcomes among preterm twins admitted to neonatal intensive care units are similar irrespective of chorionicity