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Context: Congenital hypopituitarism is a genetically heterogeneous condition. Whole exome sequencing (WES) is a promising approach for molecular diagnosis of patients with this condition. Objectives: To conduct WES in a patient with congenital hypopituitarism born to consanguineous parents, CDH2 screening in a cohort of patients with congenital hypopituitarism, and functional testing of a novel CDH2 variant. Design: Genomic DNA from a proband and her consanguineous parents was analyzed by WES. Copy number variants were evaluated. The genetic variants were filtered for population frequency (ExAC, 1000 genomes, gnomAD and ABraOM), in silico prediction of pathogenicity, and gene expression in pituitary and/or hypothalamus. Genomic DNA from 145 patients was screened for CDH2 by Sanger sequencing. Results: One female patient with deficiencies in GH, TSH, ACTH, LH, and FSH and ectopic posterior pituitary gland contained a rare homozygous c.865G>A (p.Val289Ile) variant in CDH2. To determine whether the p.Val289Ile variant in CDH2 affects cell adhesion properties, we stably transfected L1 fibroblast lines, labeled the cells with lipophilic dyes, and quantified aggregation. Large aggregates formed in cells expressing wild type CDH2, but aggregation was impaired in cells transfected with variant CDH2 or non-transfected. Conclusion: A homozygous CDH2 allelic variant was found in 1 hypopituitarism patient, and the variant impaired cell aggregation function in vitro. No disease-causing variants were found in 145 other patients screened for CDH2 variants. Thus, CDH2 is a candidate gene for hypopituitarism that needs to be tested in different populations.

The Neotropical region hosts 4225 freshwater fish species, ranking first among the world's most diverse regions for freshwater fishes. Our NEOTROPICAL FRESHWATER FISHES data set is the first to produce a large-scale Neotropical freshwater fish inventory, covering the entire Neotropical region from Mexico and the Caribbean in the north to the southern limits in Argentina, Paraguay, Chile, and Uruguay. We compiled 185,787 distribution records, with unique georeferenced coordinates, for the 4225 species, represented by occurrence and abundance data. The number of species for the most numerous orders are as follows: Characiformes (1289), Siluriformes (1384), Cichliformes (354), Cyprinodontiformes (245), and Gymnotiformes (135). The most recorded species was the characid Astyanax fasciatus (4696 records). We registered 116,802 distribution records for native species, compared to 1802 distribution records for nonnative species. The main aim of the NEOTROPICAL FRESHWATER FISHES data set was to make these occurrence and abundance data accessible for international researchers to develop ecological and macroecological studies, from local to regional scales, with focal fish species, families, or orders. We anticipate that the NEOTROPICAL FRESHWATER FISHES data set will be valuable for studies on a wide range of ecological processes, such as trophic cascades, fishery pressure, the effects of habitat loss and fragmentation, and the impacts of species invasion and climate change. There are no copyright restrictions on the data, and please cite this data paper when using the data in publications.

Introduction The Ames strain, a spontaneous mutant mouse of the Prop1 gene, is deficient in GH, TSH, PRL and gonadotropins. In recent years, it has been the focus of studies on aging and there is a growing interest in elucidating the role of hormone replacement (HR) in the hypothalamic-pituitary-gonadal (HPG) axis. Aim The present study aimed to characterize sexual maturation and fertility restoration in isogenic strain under or not HR. Material and methods Five homozygous male animals were treated via intraperitoneal injections, starting 30 days postnatally, with levothyroxine 3x/week and recombinant human GH 5x/week for 40 days, followed by maintenance applications of both hormones once a week until they complete 90 days post natal. The sexual maturity was evaluated looking for balanoprepucial opening and descendent testis comparing untreated homozygous (n=5) with their wild type siblings (n=5). Fertility was evaluated by mating known fertile wild type animals with treated and untreated animals for 8 times. Reproductive parameters were evaluated between groups using histological sections of the testes (HE) and classified according to johnsen score where 1-2 was classified as absent germline cells and 8-10 normal development. At the sacrifice, testicular weight was measured using scale and spermogram was done looking for motility and viability in the sperm collected in the seminiferous tube. Gata2 transcription factor and pituitary hormones Lh, Fsh, Tsh, Prl and Gh transcriptional analysis were performed by RT-qPCR. Results The homozygous treated animals presented a 2 weeks delay in the age of sexual maturation compared to wild animals. Their fertility and reproductive parameters were restored noticed by increased testicular weight, improved spermatogenesis, similar morphology of seminiferous tubules (johnsen score 8.7) and spermogram compared to the wild type, besides presenting offspring when mated to their wild siblings. Sexual maturity was absent in most of the untreated homozygous animals presenting no offspring. The reproductive parameters in the untreated homozygous animals presented reduced testicular weight, size of seminiferous tubes leading to johnsen score as 5 and an azospermic spermogram in all animals. Interestingly 2 untreated homozugous animal had spontaneous maturation. Gata2 was significant decreased in the untreated animal (0,49 ± 0,19) compared to wild type (1 ± 0,15) (p≤ 0.019). LH transcriptional pattern was significant increase in the animals under treatment (1.56 ± 0.68) compared to homozygotes without hormonal intervention (0.36 ± 0.34) (p≤ 0.048). Conclusion Ames mutant mice under treatment with GH and levothyroxine replacement reached sexual maturation and restored fertility and the mechanics behind this phenomenon will be explored using RNA seq in the future. Presentation: Sunday, June 12, 2022 11:45 a.m. - 12:00 p.m.

The zebrafish (Danio rerio) is an emerging model system in several research areas worldwide, especially in the Global South. In this context, the present study revised the historical use and trends of zebrafish as experimental models in Brazil. The data concerning the bibliometric parameters, research areas, geographic distribution, experimental design, zebrafish strain, and reporter lines, as well as recent advances were revised. In addition, the comparative trends of Brazilian and global research were discussed. Revised data showed the rapid growth of Brazilian scientific production using zebrafish as a model, especially in three main research areas (Neuroscience &and Behavior, Pharmacology and Toxicology, and Environment/Ecology). Studies were conducted in 19 Brazilian states (70.37%), confirming the wide geographic distribution and importance of zebrafish research. Results indicated that research related to toxicological approaches are widespread in Global South countries such as Brazil. Studies were performed mainly using in vivo tests (89.58%) with adult fish (59.75%) and embryos (30.67%). Moreover, significant research gaps and recommendations for future research are presented. The present study shows that the zebrafish is a suitable vertebrate model system in the Global South.

Iron oxide nanoparticles (IONPs) have been tested to remediate aquatic environments polluted by chemicals, such as pesticides. However, their interactive effects on aquatic organisms remain unknown. This study aimed to investigate the genotoxicity and mutagenicity of co-exposure of IONPs (γ-Fe2O3 NPs) and glyphosate-based herbicide (GBH) in the fish Poecilia reticulata. Thus, fish were exposed to citrate-functionalized γ-Fe2O3 NPs (0.3 mg L-1; 5.44 nm) alone or co-exposed to γ-Fe2O3 NPs (0.3 mg L-1) and GBH (65 and 130 μg of glyphosate L-1) during 14 and 21 days. The genotoxicity (DNA damage) was analyzed by comet assay, while the mutagenicity evaluated by micronucleus test (MN test) and erythrocyte nuclear abnormalities (ENA) frequency. The co-exposure induced clastogenic (DNA damage) and aneugenic (nuclear alterations) effects on guppies in a time-dependent pattern. Fish co-exposed to NPs and GBH (130 μg glyphosate L-1) showed high DNA damage when compared to NPs alone and control group, indicating synergic effects after 21 days of exposure. However, mutagenic effects (ENA) were observed in the exposure groups after 14 and 21 days. Results showed the potential genotoxic and mutagenic effects of maghemite NPs and GBH co-exposure to freshwater fish. The transformation and interaction of iron oxide nanoparticles with other pollutants, as herbicides, in the aquatic systems are critical factors in the environmental risk assessment of metal-based NPs.

The iron oxide nanoparticles (IONPs) have been widely applied to nanomedicine, biology, and nanoremediation. However, its ecotoxicological effects in aquatic organisms remain unclear. The guppy (Poecilia reticulata) is a suitable model to assess the ecotoxicity, reproductive toxicity, and environmental risk of emergent pollutants, such as nanoparticles (NPs). In this study, the effects of citrate-functionalized maghemite (γ-Fe2O3) NPs on reproductive behavior and spermatogenesis of guppies were investigated over 21 days of exposure. Male guppies were exposed to maghemite NPs at environmentally relevant concentrations (0.3 and 3.0 μg L−1) for 14 and 21 days, jointly with a control group (reconstituted water). The reproductive toxicity was analyzed in terms of biometric parameters, sexual behavior, and gonadal histopathology (qualitative and histomorphometric analyzes following by histopathological indices). Results indicated that the chronic exposure to maghemite NPs did not affect male reproductive behavior, but changed the spermatogenesis (increased the spermatid S1 and decreased the spermatocytes and spermatozeugmata frequency). The maghemite NPs induced histopathological damages associated with inflammatory responses and regressive alterations in the testis of guppies. The IONPs can lead to reproductive impairment in freshwater fish, indicating its potential environmental risk.

Water safety is a world-wide concern and several efforts have been made in order to ensure the conservation of aquatic ecosystems. Water quality monitoring must be performed with an integrated approach using biomonitor organisms allied to water parameters. Nonetheless, very few studies have focused on biomarker responses in neotropical fish, especially in the freshwater ecosystem of Brazilian Cerrado savanna. In present study, the active biomonitoring of the João Leite river (central Brazilian Cerrado river) was performed through the evaluation of biomarker responses in caged Astyanax lacustris in combination with land use classification and analysis of water parameters. Caged fish were exposed for seven days at four sites along the river and two control groups were kept in a tank under controlled conditions. Results showed that pasture was the predominant land use in the João Leite river basin (54.07%), followed by natural vegetation (34.92%) and other kind of land use (11.01%). Water analyses showed metal concentrations (Mn and Fe) above the maximum allowed by Brazilian regulation, with particularly higher concentrations at Site 2 (near to pasture area). Biomarker responses did not show significant differences for somatic and mutagenic biomarkers between sites. However, the comet assay showed high DNA damage at Sites 2 and 3, indicating genotoxic effects in caged fish at pasture areas. Histopathological analysis showed highest frequency of leukocyte infiltration in liver of fish from Site 2, confirming the ecotoxic effects on A. lacustris in streams impacted by grazing activities. DNA damage and leukocyte infiltration in fish hepatic tissues were sensitive biomarkers in the neotropical fish A. lacustris to assess the environment health of the Cerrado river. These results showed the importance of using a multibiomarker approach in environmental risk assessment, especially in areas more at risk from anthropogenic pollution.