Mutations in SRY and WT1 genes required for gonadal development are not responsible for XY partial gonadal dysgenesis

The WT1 transcription factor regulates SRY expression during the initial steps of the sex determination process in humans, activating a gene cascade leading to testis differentiation. In addition to causing Wilms' tumor, mutations in WT1 are often responsible for urogenital defects in men, while SRY mutations are mainly related to 46,XY pure gonadal dysgenesis. In order to evaluate their role in abnormal testicular organogenesis, we screened for SRY and WT1 gene mutations in 10 children with XY partial gonadal dysgenesis, 2 of whom with a history of Wilms' tumor. The open reading frame and 360 bp of the 5' flanking sequence of the SRY gene, and the ten exons and intron boundaries of the WT1 gene were amplified by PCR of genomic DNA. Single-strand conformation polymorphism was initially used for WT1 mutation screening. Since shifts in fragment migration were only observed for intron/exon 4, the ten WT1 exons from all patients were sequenced manually. No mutations were detected in the SRY 5' untranslated region or within SRY open-reading frame sequences. WT1 sequencing revealed one missense mutation (D396N) in the ninth exon of a patient who also had Wilms' tumor. In addition, two silent point mutations were found in the first exon including one described here for the first time. Some non-coding sequence variations were detected, representing one new (IVS4+85A>G) and two already described (-7ATG T>G, IVS9-49 T>C) single nucleotide polymorphisms. Therefore, mutations in two major genes required for gonadal development, SRY and WT1, are not responsible for XY partial gonadal dysgenesis.172

Morphometry And Histology Of Gonads From 13 Children With Dysgenetic Male Pseudohermaphroditism

Background. - Dysgenetic male pseudohermaphroditism (DMP) is a sexual differentiation disorder characterized by bilateral dysgenetic testes, persistent müllerian structures, and cryptorchidism in individuals with a 46,XY karyotype. However, the histologic criteria for the diagnosis of DMP are poorly established. Objective. - To determine gonadal histology in children with DMP. Patients and Methods. - Between 1996 and 1998, 13 patients with DMP were evaluated on our service. The clinical diagnosis of DMP was based on a 46,XY karyotype, sex ambiguity, high levels of follicle-stimulating hormone and low levels Of antimüllerian hormone, a decreased testosterone response to human chorionic gonadotropin stimulation without accumulation of testosterone precursors, and the presence of müllerian structures. Molecular sequencing the HMGbox region of the SRY gene did not reveal any mutations. Biopsies were performed for 22 of 26 gonads (patient age at the time of biopsy, 16 months to 10 years). Conventional microscopy was used to evaluate mean tubular diameter, tubular fertility index, and number of Sertoli cells per tubular profile. Results. - All 26 gonads were located outside of the labioscrotal folds. Their histologic features varied from only a reduction in tubular size to features of a streak gonad. Five of the 22 gonads grossly resembled a streak gonad. The mean tubular diameter was severely reduced (>30% reduction relative to the normal tubular diameter for the patient's age) in 4 gonads, markedly reduced (10%-30%) in 11 gonads, slightly reduced (<10%) in one gonad, and normal in one gonad. The tubular fertililty index, expressed as the percentage of tubular profiles containing germ cells, was severely reduced (<30% of normal values) in 9 gonads, markedly reduced (50%-30%) in 2 gonads, and normal in 6 gonads. The number of Sertoli cells per tubular profile was elevated in 16 gonads and normal in one gonad. Thin tubules surrounded by fibrous tissue were occasionally observed. Conclusion. - The histologic findings confirmed the clinical diagnosis of DMP in every patient in the present series. However, gonadal histology was variable, and careful morphometric evaluation may be necessary to establish the diagnosis.1255652656Robboy, S.J., Miller, T., Donahoe, P.K., Dysgenesis of testicular and streak gonads in the syndrome of mixed gonadal dysgenesis: Perspective derived from clinicopathologic analysis of twenty-one cases (1982) Hum Pathol, 13, pp. 700-716Troche, V., Hernandez, E., Neoplasia arising in dysgenetic gonads (1986) Obstet Gynecol Surv, 41, pp. 74-79Krasna, I.H., Lee, M.L., Smilow, P., Sciorra, L., Eierman, L., Risk of malignancy in bilateral streak gonads: The role of the Y chromosome (1992) J Pediatr Surg, 27, pp. 1376-1380Rohatgi, M., Gupta, D.K., Menon, P.S., Verma, I.C., Mathur, M., Mixed gonadal dysgenesis and dysgenetic male pseudohermaphroditism - A critical analysis (1992) Indian J Pediatr, 59, pp. 487-500Rey, R.A., Belville, C., Nhou-Fékété, C., Evaluation of gonadal function in 107 intersex patients by means of serum antimüllerian hormone measurement (1999) J Clin Endocrinol Metab, 84, pp. 627-631Stuchi-Perez, E.G., Lukas-Croisier, C., Castro, M., Evaluation of the tubular and interstitial functions of the testis in 46,XY patients with ambiguous genitalia (2000) J Pediatr Endocrinol Metab, 13, pp. 605-612Chang, H.J., Clark, R.D., Bachman, H., The phenotype of 45,X/46,XY mosaicism: An analysis of 92 prenatally diagnosed cases (1990) Am J Hum Genet, 46, pp. 156-167Rajfer, J., Walsh, P.C., Mixed gonadal dysgenesis: Dysgenetic male pseudoher-maphroditism (1981) The Intersex Child: Pediatric and Adolescent Endocrinology, pp. 103-115. , Josso N, ed. Basel, Switzerland: S. KargerBorer, J.G., Nitti, V.W., Glassberg, K.I., Mixed gonadal dysgenesis and dysgenetic male pseudohermaphroditism (1995) J Urol, 153, pp. 1267-1273Donahoe, P.K., Crawford, J.D., Hendren, W.H., Mixed gonadal dysgenesis: Pathogenesis and management (1979) J Pediatr Surg, 14, pp. 287-300Pelletier, J., Bruening, W., Kashtan, C.E., Germline mutations in the Wilms' tumor supressor gene are associated with abnormal urogenital development in Denys-Drash syndrome (1991) Cell, 67, pp. 437-1147Carré-Éusebe, D., Imbeaud, S., Harbison, M., New, M.I., Josso, N., Picard, J.Y., Variants of the anti-Müllerian hormone gene in a compound heterozygote with the persistent Müllerian duct syndrome and his family (1992) Hum Genet, 90, pp. 389-394Nistal, M., Paniagua, R., Non-neoplastic diseases of the testis (1996) Urologic Surgical Pathology, pp. 458-565. , Bostiwick DG, Eble JN, eds. St Louis, Mo: MosbyLennox, B., Ahmad, K.M., Mack, W.S., A method for determining the relative total length of the tubules in the testis (1970) J Pathol, 102, pp. 229-238Jimenez, R., Sanchez, A., Burgos, M., Dias De La Guardia, R.C., Puzzling out the genetics of mammalian sex determination (1996) Trends Genet, 12, pp. 164-166Müller, J., Skakkebaek, N.F., Quantification of germ cells and seminiferous tubules by stereological examination of testicles from 50 boys who suffered from sudden death (1983) Int J Androl, 6, pp. 143-156Cortes, D., Müller, J., Skakkebaek, N.E., Proliferation of Sertoli cells during development of the human testis assessed by stereological methods (1987) Int J Androl, 10, pp. 589-596Nistal, M., Abaurrea, M.A., Paniagua, R., Morphological and histometric study on the human Sertoli cell from birth to the onset of puberty (1982) J Anat, 14, pp. 351-363Van Niekerk, W.A., Retief, A.E., The gonads of human true hermaphrodites (1981) Hum Genet, 58, pp. 117-122Guerra Jr., G., De Mello, M.P., Assumpção, J.G., True hermaphrodites in the southeastern region of Brazil: A different cytogenetic and gonadal profile (1998) J Pediatr Endocrinol Metab, 11, pp. 519-524Quigley, C.A., De Bellis, A., Marschke, K.B., El-Awady, M.K., Wilson, E.M., French, F.S., Androgen receptor defects: Historical, clinical and molecular perspectives (1995) Endocr Rev, 16, pp. 271-32

Modulation of extracellular matrix by nutritional hepatotrophic factors in thioacetamide-induced liver cirrhosis in the rat

Nutritional substances associated to some hormones enhance liver regeneration when injected intraperitoneally, being denominated hepatotrophic factors (HF). Here we verified if a solution of HF (glucose, vitamins, salts, amino acids, glucagon, insulin, and triiodothyronine) can revert liver cirrhosis and how some extracellular matrices are affected. Cirrhosis was induced for 14 weeks in 45 female Wistar rats (200 mg) by intraperitoneal injections of thioacetamide (200 mg/kg). Twenty-five rats received intraperitoneal HF twice a day for 10 days (40 mL·kg-1·day-1) and 20 rats received physiological saline. Fifteen rats were used as control. The HF applied to cirrhotic rats significantly: a) reduced the relative mRNA expression of the genes: Col-&#945;1 (-53%), TIMP-1 (-31.7%), TGF-&#946;1 (-57.7%), and MMP-2 (-41.6%), whereas Plau mRNA remained unchanged; b) reduced GGT (-43.1%), ALT (-17.6%), and AST (-12.2%) serum levels; c) increased liver weight (11.3%), and reduced liver collagen (-37.1%), regenerative nodules size (-22.1%), and fibrous septum thickness. Progranulin protein (immunohistochemistry) and mRNA (in situ hybridization) were found in fibrous septa and areas of bile duct proliferation in cirrhotic livers. Concluding, HF improved the histology and serum biochemistry of liver cirrhosis, with an important reduction of interstitial collagen and increased extracelullar matrix degradation by reducing profibrotic gene expression

Unjamming of active rotators

Active particle assemblies can exhibit a wide range of interesting dynamical phases depending on internal parameters such as density, adhesion strength or self-propulsion. Active self-rotations are rarely studied in this context, although they can be relevant for active matter systems, as we illustrate by analyzing the motion of Chlamydomonas reinhardtii algae under different experimental conditions. Inspired by this example, we simulate the dynamics of a system of interacting active disks endowed with active torques and self-propulsive forces. At low packing fractions, adhesion causes the formation of small rotating clusters, resembling those observed when algae are stressed. At higher densities, the model shows a jamming to unjamming transition promoted by active torques and hindered by adhesion. We also study the interplay between self-propulsion and self-rotation and derive a phase diagram. Our results yield a comprehensive picture of the dynamics of active rotators, providing useful guidance to interpret experimental results in cellular systems where rotations might play a role

Acolhimento de refugiados: alimentação e necessidades nutricionais em situações de emergência

A Europa é neste momento um dos principais destinos de um intenso fluxo migratórioprovocado por diferentes conflitos armados no Médio Oriente e em África, tendo a ComissãoEuropeia (CE) acordado na distribuição de uma parte destas pessoas em clara necessidade deproteção nacional, pelos diversos Estados Membros. As populações em trânsito e ascaracterísticas do seu acolhimento possuem especificidades que podem comprometer oacesso a uma alimentação adequada e a cuidados básicos de saúde, influenciando amorbilidade e a mortalidade nos grupos afetados.Este manual pretende estabelecer um referencial para a intervenção alimentar e nutricionalaos refugiados que chegam a Portugal. Destina-se a todos aqueles que prestam apoio, quer anível individual, quer a nível institucional, e que sejam responsáveis por qualquer aspetorelacionado com a saúde e a alimentação dessas populações, facilitando a operacionalizaçãoda assistência e fornecendo ferramentas para a tomada de decisões.O manual está organizado em 3 partes. Inicia-se com a avaliação do estado nutricional dapopulação a acolher, apresenta depois diversas estratégias para o desenho da intervençãoalimentar e nutricional tendo por base as necessidades nutricionais previstas para estes grupospopulacionais e, na parte final, aborda a importância de garantir a higiene e segurança dosalimentos na prestação desta assistência alimentar. Este manual apresenta ainda algumasconsiderações relacionadas com os cuidados básicos na área da psicologia destinados àsequipas que estão no terrenoEurope is now a major destination of an intense migratory flow caused by armed conflicts inthe Middle East and Africa. In this sense, the European Commission (EC) agreed on thedistribution of some of these people in clear need of international protection by variousMember States. The populations in transit and the characteristics of its reception havespecificities that can compromise the access to adequate food and basic health care. They canalso influence morbidity and mortality in affected groups.This manual aims to establish a framework of nutrition and food intervention to refugees inPortugal. It is aimed at all those who support it, either individually or institutionally, and areresponsible for any aspect related to the health and nutrition of these populations, facilitatingthe implementation of care and providing tools for decision making. The manual is organizedinto 3 parts. It begins with the assessment of nutritional status of the population. Then presents several strategies to support the design of food and nutrition intervention based onthe estimated nutritional needs for these population groups. In the final section it addressesthe importance of ensuring food safety and hygiene when providing this food assistance. Thismanual also addresses some considerations towards basic psychological interventionprinciples, destined to help the teams on the field

SpectraNet–53: A deep residual learning architecture for predicting soluble solids content with VIS–NIR spectroscopy

This work presents a new deep learning architecture, SpectraNet-53, for quantitative analysis of fruit spectra, optimized for predicting Soluble Solids Content (SSC, in Brix). The novelty of this approach resides in being an architecture trainable on a very small dataset, while keeping a performance level on-par or above Partial Least Squares (PLS), a time-proven machine learning method in the field of spectroscopy. SpectraNet-53 performance is assessed by determining the SSC of 616 Citrus sinensi L. Osbeck 'Newhall' oranges, from two Algarve (Portugal) orchards, spanning two consecutive years, and under different edaphoclimatic conditions. This dataset consists of short-wave near-infrared spectroscopic (SW-NIRS) data, and was acquired with a portable spectrometer, in the visible to near infrared region, on-tree and without temperature equalization. SpectraNet-53 results are compared to a similar state-of-the-art architecture, DeepSpectra, as well as PLS, and thoroughly assessed on 15 internal validation sets (where the training and test data were sampled from the same orchard or year) and on 28 external validation sets (training/test data sampled from different orchards/years). SpectraNet-53 was able to achieve better performance than DeepSpectra and PLS in several metrics, and is especially robust to training overfit. For external validation results, on average, SpectraNet-53 was 3.1% better than PLS on RMSEP (1.16 vs. 1.20 Brix), 11.6% better in SDR (1.22 vs. 1.10), and 28.0% better in R2 (0.40 vs. 0.31).project NIBAP ALG-01-0247-FEDER-037303, project OtiCalFrut ALG-010247-FEDER-033652info:eu-repo/semantics/publishedVersio

Implante transcateter valve‐in‐valve para disfunção de biopróteses cirúrgicas aórticas

RESUMOIntroduçãoEstudos recentes têm demonstrado a eficácia do implante transcateter valve‐in‐valve para o tratamento de disfunção de biopróteses em pacientes de alto risco cirúrgico. Apresentamos nossa experiência inicial com o implante valve‐in‐valve.MétodosCaracterizamos o perfil clínico, ecocardiográfico e do procedimento, e reportamos os resultados de médio prazo de pacientes com disfunção de bioprótese submetidos a implante valve‐in‐valve em posição aórtica.ResultadosIncluímos sete pacientes do sexo masculino, com idade de 72,6 ± 10,0 anos. O escore STS foi 9,6 ± 10,5%, e o EuroSCORE logístico foi 22,7 ± 14,7%. Três pacientes apresentavam dupla disfunção; dois tinham insuficiência; e dois exibiam estenose isolada. A via transfemoral foi utilizada em seis casos, e a transapical, em um caso. Os dispositivos implantados incluíram as próteses Sapien XT (n = 5) e CoreValve (n = 2). O sucesso do procedimento foi obtido em seis (85,7%) casos. Após o procedimento, o gradiente médio reduziu‐se de 38,2 ± 9,6mmHg para 20,9 ± 5,9mmHg, e a área valvar elevou‐se de 1,2 ± 0,4cm2 para 1,5 ± 0,5cm2. Ao final de 1 ano, não ocorreram óbitos e nem outros desfechos adversos significativos; 80% dos pacientes encontravam‐se em classe funcional NYHA I/II. Os gradientes transvalvares e a área valvar permaneceram inalterados nesse período.ConclusõesO procedimento valve‐in‐valve foi eficaz na maioria dos pacientes de alto risco cirúrgico com disfunção de bioprótese. Quando realizado em pacientes bem selecionados, resulta em desfechos clínicos e hemodinâmicos satisfatórios.ABSTRACTBackgroundRecent studies have demonstrated the efficacy of the transcatheter valve‐in‐valve implantation for the treatment of bioprosthesis dysfunction in high‐risk surgical patients. This study presents the initial experience with valve‐in‐valve implantation.MethodsClinical, echocardiographic, and procedural profiles were characterized, and the mid‐term results of patients with surgical bioprosthesis dysfunction submitted to valve‐in‐valve implantation in the aortic position were reported.ResultsSeven male patients were included, aged 72.6 ± 10.0 years. The STS score was 9,6 ± 10,5%, and the logistic EuroSCORE was 22.7 ± 14.7%. Three patients had combined aortic bioprosthesis failure; two had isolated regurgitation; and two had isolated stenosis. The transfemoral access was used in six cases, and the transapical access in one case. Implanted devices included Sapien XT (n = 5) and CoreValve (n = 2) prostheses. Procedural success was achieved in six (85.7%) cases. After the procedure, the mean gradient decreased from 38.2 ± 9.6mmHg to 20.9 ± 5.9mmHg, and the valve area increased from 1.2 ± 0.4cm2 to 1.5 ± 0.5cm2. After 1 year, there were no deaths and no other significant adverse outcomes; 80% of patients were in NYHA functional class I/II. The transvalvular gradients and valve area remained unchanged in this period.ConclusionsThe valve‐in‐valve procedure was effective in most high‐risk surgical patients with bioprosthesis dysfunction. When performed in well‐selected patients, it results in satisfactory clinical and hemodynamic outcomes

Resistenza di Pseudomonas aeruginosa a ciprofloxacina e levofloxacina : 1998-2002

The aim of this study was to evaluate resistance rates to ciprofloxacin and levofloxacin of Pseudomonas aeruginosa (n=1917 strains) isolated at Laboratory of Microbiology at L. Sacco Teaching Hospital in Milan, Italy in the period between January 1998 and October 2002. Twenty-five percent of tested strains were isolated from sputum, 18% from bronchial lavage, 10% from urine, 9% from ear, 4.5% from blood and 26% from other materials. Ciprofloxacin-resistant strains were 121/ 411 (29%) in 1998, 158/ 526 (30%) in 1999, 136/ 400 (34%) in 2000, 129/390 (33%) in 2001 and 53/ 190 (28%) in 2002. Resistance rates for levofloxacin were 132/472 (28%) in 1999, 104/ 400 (26%) in 2000, 101/ 390 (26 %) in 2001 and 47/ 190 (25%) in 2002. Our data highlight overall stability in resistance to ciprofloxacin and levofloxacin with minor variations for ciprofloxacin. Moreover, in contrast with international worldwide studies, resistance rates to ciprofloxacin remained higher than those related to levofloxacin throughout the period studied

On the freezing of variables in random constraint satisfaction problems

The set of solutions of random constraint satisfaction problems (zero energy groundstates of mean-field diluted spin glasses) undergoes several structural phase transitions as the amount of constraints is increased. This set first breaks down into a large number of well separated clusters. At the freezing transition, which is in general distinct from the clustering one, some variables (spins) take the same value in all solutions of a given cluster. In this paper we study the critical behavior around the freezing transition, which appears in the unfrozen phase as the divergence of the sizes of the rearrangements induced in response to the modification of a variable. The formalism is developed on generic constraint satisfaction problems and applied in particular to the random satisfiability of boolean formulas and to the coloring of random graphs. The computation is first performed in random tree ensembles, for which we underline a connection with percolation models and with the reconstruction problem of information theory. The validity of these results for the original random ensembles is then discussed in the framework of the cavity method.Comment: 32 pages, 7 figure