Thyroid hormones T3 and T4 regulate human luteinized granulosa cells, counteracting apoptosis and promoting cell survival.

PURPOSE: Fine and balanced regulation of cell proliferation and apoptosis are key to achieve ovarian follicle development from the primordial to the preovulatory stage and therefore assure female reproductive function. While gonadotropins are the major and most recognized regulators of follicle cell growth and function, other factors, both systemic and local, play equally important roles. This work is aimed at evaluating the effects of thyroid hormones (THs) on human granulosa luteinized (hGL) viability. METHODS: Human GL cells derived from assisted reproduction treatments were exposed to T3 or T4. Cell viability was evaluated by MTT assay. Apoptosis was evaluated by the TUNEL assay and active caspase-3 staining. StAR, CYP19A1,Caspase-3, P53 and BAX mRNA were evaluated by real-time PCR. LC3-I/-II, AKT and pAKT were evaluated by western blot. RESULTS: T3 and T4 promoted cell viability in a dose-dependent modality and modulate StAR and CYP19A1 expression. T3 and to a lesser extent T4 mitigated cell death induced by serum starvation by inhibition of caspase-3 activity and expression of P53 and BAX; and attenuate cell death experimentally induced by C2-ceramide. Cell death derived from starvation appeared to be involved in autophagic processes, as the levels of autophagic markers (LC3-II/LC3-I ratio) decreased when starved cells were exposed to T3 and T4. This effect was associated with an increase in pAkt levels. CONCLUSION: From the present study, THs emerge as potent anti-apoptotic agents in hGL cells. This effect is achieved by inhibiting the apoptosis signalling pathway of BAX and caspase-3, while maintaining active the PI3K/AKT pathway.

Preliminar results of paleontological salvage at Belo Monte Powerplant construction.

In this paper some preliminary fossil specimens are presented. They represent a collection sampled by Belo Monte's Programa de Salvamento do Patrimônio Paleontológico (PSPP), which includes unprecedented invertebrate fauna and fossil vertebrates from Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão formations from Amazonas basin, Brazil. The Belo Monte paleontological salvage was able to recover 495 microfossil samples and 1744 macrofossil samples on 30 months of sampling activities, and it is still ongoing. The macrofossils identified are possible plant remains, ichnofossils, graptolites, brachiopods, molluscs, athropods, Agnatha, palynomorphs (miosphores, acritarchs, algae cysts, fungi spores and unidentified types) and unidentified fossils. However, deep scientific research is not part of the scope of the program, and this collection must be further studied by researchers who visit Museu Paraense Emilio Goeldi, where the fossils will be housed. More material will be collected until the end of the program. The collection sampled allows a mosaic composition with the necessary elements to assign, in later papers, taxonomic features which may lead to accurate species identification and palaeoenvironmental interpretations.

Preliminar results of paleontological salvage at Belo Monte Powerplant construction

Abstract In this paper some preliminary fossil specimens are presented. They represent a collection sampled by Belo Montes Programa de Salvamento do Patrimônio Paleontológico (PSPP), which includes unprecedented invertebrate fauna and fossil vertebrates from Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão formations from Amazonas basin, Brazil. The Belo Monte paleontological salvage was able to recover 495 microfossil samples and 1744 macrofossil samples on 30 months of sampling activities, and it is still ongoing. The macrofossils identified are possible plant remains, ichnofossils, graptolites, brachiopods, molluscs, athropods, Agnatha, palynomorphs (miosphores, acritarchs, algae cysts, fungi spores and unidentified types) and unidentified fossils. However, deep scientific research is not part of the scope of the program, and this collection must be further studied by researchers who visit Museu Paraense Emilio Goeldi, where the fossils will be housed. More material will be collected until the end of the program. The collection sampled allows a mosaic composition with the necessary elements to assign, in later papers, taxonomic features which may lead to accurate species identification and palaeoenvironmental interpretations.

Resumo Neste artigo são apresentados de forma preliminar alguns espécimes fósseis representativos do acervo coletado no Programa de Salvamento do Patrimônio Paleontológico (PSPP) da UHE Belo Monte, que inclui fauna vertebrada e invertebrada inédita das formações Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão, da bacia do Amazonas, Brasil. No salvamento paleontológico de Belo Monte foi possível recuperar 495 amostras para microfósseis e 1744 amostras com macrofósseis em 30 meses de amostragem, que ainda continua em andamento. Os macrofósseis identificados são possíveis restos de plantas, icnofósseis, graptolitos, braquiópodes, moluscos, artópodes, Agnatha, palinomorfos (miósporos, acritarcas, cistos de alga, esporos de fungo e tipos indeterminados) além de fósseis não identificados. No entanto, pesquisa científica aprofundada não é parte do escopo do programa, e esta coleção deve ser estudada no futuro por pesquisadores pela visita ao Museu Paraense Emilio Goeldi, onde os fósseis ficarão depositados. Mais material será coletado até o fim de 2015. A assemblagem coletada permite a composição de mosaico com elementos necessários para a definição, em artigos futuros, características taxonômicas que levarão à identificação precisa de espécies e interpretações paleoambientais.

Preliminar results of paleontological salvage at Belo Monte Powerplant construction / Resultados preliminares do programa de salvamento paleontológico da construção da Hidrelétrica de Belo Monte

In this paper some preliminary fossil specimens are presented. They represent a collection sampled by Belo Monte’s Programa de Salvamento do Patrimônio Paleontológico (PSPP), which includes unprecedented invertebrate fauna and fossil vertebrates from Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão formations from Amazonas basin, Brazil. The Belo Monte paleontological salvage was able to recover 495 microfossil samples and 1744 macrofossil samples on 30 months of sampling activities, and it is still ongoing. The macrofossils identified are possible plant remains, ichnofossils, graptolites, brachiopods, molluscs, athropods, Agnatha, palynomorphs (miosphores, acritarchs, algae cysts, fungi spores and unidentified types) and unidentified fossils. However, deep scientific research is not part of the scope of the program, and this collection must be further studied by researchers who visit Museu Paraense Emilio Goeldi, where the fossils will be housed. More material will be collected until the end of the program. The collection sampled allows a mosaic composition with the necessary elements to assign, in later papers, taxonomic features which may lead to accurate species identification and palaeoenvironmental interpretations.

Neste artigo são apresentados de forma preliminar alguns espécimes fósseis representativos do acervo coletado no Programa de Salvamento do Patrimônio Paleontológico (PSPP) da UHE Belo Monte, que inclui fauna vertebrada e invertebrada inédita das formações Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão, da bacia do Amazonas, Brasil. No salvamento paleontológico de Belo Monte foi possível recuperar 495 amostras para microfósseis e 1744 amostras com macrofósseis em 30 meses de amostragem, que ainda continua em andamento. Os macrofósseis identificados são possíveis restos de plantas, icnofósseis, graptolitos, braquiópodes, moluscos, artópodes, Agnatha, palinomorfos (miósporos, acritarcas, cistos de alga, esporos de fungo e tipos indeterminados) além de fósseis não identificados. No entanto, pesquisa científica aprofundada não é parte do escopo do programa, e esta coleção deve ser estudada no futuro por pesquisadores pela visita ao Museu Paraense Emilio Goeldi, onde os fósseis ficarão depositados. Mais material será coletado até o fim de 2015. A assemblagem coletada permite a composição de mosaico com elementos necessários para a definição, em artigos futuros, características taxonômicas que levarão à identificação precisa de espécies e interpretações paleoambientais.

Preliminar results of paleontological salvage at Belo Monte Powerplant construction / Resultados preliminares do programa de salvamento paleontológico da construção da Hidrelétrica de Belo Monte

Abstract In this paper some preliminary fossil specimens are presented. They represent a collection sampled by Belo Monte’s Programa de Salvamento do Patrimônio Paleontológico (PSPP), which includes unprecedented invertebrate fauna and fossil vertebrates from Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão formations from Amazonas basin, Brazil. The Belo Monte paleontological salvage was able to recover 495 microfossil samples and 1744 macrofossil samples on 30 months of sampling activities, and it is still ongoing. The macrofossils identified are possible plant remains, ichnofossils, graptolites, brachiopods, molluscs, athropods, Agnatha, palynomorphs (miosphores, acritarchs, algae cysts, fungi spores and unidentified types) and unidentified fossils. However, deep scientific research is not part of the scope of the program, and this collection must be further studied by researchers who visit Museu Paraense Emilio Goeldi, where the fossils will be housed. More material will be collected until the end of the program. The collection sampled allows a mosaic composition with the necessary elements to assign, in later papers, taxonomic features which may lead to accurate species identification and palaeoenvironmental interpretations.

Resumo Neste artigo são apresentados de forma preliminar alguns espécimes fósseis representativos do acervo coletado no Programa de Salvamento do Patrimônio Paleontológico (PSPP) da UHE Belo Monte, que inclui fauna vertebrada e invertebrada inédita das formações Pitinga, Jatapu, Manacapuru, Maecuru e Alter do Chão, da bacia do Amazonas, Brasil. No salvamento paleontológico de Belo Monte foi possível recuperar 495 amostras para microfósseis e 1744 amostras com macrofósseis em 30 meses de amostragem, que ainda continua em andamento. Os macrofósseis identificados são possíveis restos de plantas, icnofósseis, graptolitos, braquiópodes, moluscos, artópodes, Agnatha, palinomorfos (miósporos, acritarcas, cistos de alga, esporos de fungo e tipos indeterminados) além de fósseis não identificados. No entanto, pesquisa científica aprofundada não é parte do escopo do programa, e esta coleção deve ser estudada no futuro por pesquisadores pela visita ao Museu Paraense Emilio Goeldi, onde os fósseis ficarão depositados. Mais material será coletado até o fim de 2015. A assemblagem coletada permite a composição de mosaico com elementos necessários para a definição, em artigos futuros, características taxonômicas que levarão à identificação precisa de espécies e interpretações paleoambientais.

The effect of hepatocyte growth factor on the initial stages of mouse follicle development.

Interactions between theca and granulosa cells of the follicle are critical for the coordination of ovarian follicle development. The cell-cell interactions are mediated through the local production and actions of a variety of factors. The current study is designed to investigate the expression of Hgf and its receptor, c-Met, in the mouse ovary during in vivo folliculogenesis. We found that Hgf and c-Met mRNAs were already expressed in 2-day-old ovaries, and that, while c-Met levels remained constant until 22-day-old, Hgf levels slightly but not significantly increased with age. The expression of Hgf mRNA in theca/interstitial cells was higher than in granulosa cells in 22-day-old ovaries. Immunohistochemistry analysis confirmed the expression pattern demonstrated by RT-PCR. We investigated the role of hepatocyte growth factor (HGF) at the beginning of mouse folliculogenesis and its possible interaction with kit ligand (KL). Interestingly, both KL and HGF were able to increase the expression of each other, creating a positive feedback loop. In the presence of HGF, we observed an increase of granulosa cell proliferation and an increase in the number of pre-antral and early antral follicles in ovary organ cultures. We also observed a significant increase in the diameters of follicles in individual follicle cultures. Moreover, HGF stimulated the expression of the FSH receptors, both in the whole ovary and in isolated pre-antral follicle cultures. Based on the data presented, we concluded that HGF exerts multiple levels of control over follicular cell functions, which collectively enable the progression of follicular development.

Targeting of a tail-anchored protein to endoplasmic reticulum and mitochondrial outer membrane by independent but competing pathways.

Many mitochondrial outer membrane (MOM) proteins have a transmembrane domain near the C terminus and an N-terminal cytosolic moiety. It is not clear how these tail-anchored (TA) proteins posttranslationally select their target, but C-terminal charged residues play an important role. To investigate how discrimination between MOM and endoplasmic reticulum (ER) occurs, we used mammalian cytochrome b(5), a TA protein existing in two, MOM or ER localized, versions. Substitution of the seven C-terminal residues of the ER isoform or of green fluorescent protein reporter constructs with one or two arginines resulted in MOM-targeted proteins, whereas a single C-terminal threonine caused promiscuous localization. To investigate whether targeting to MOM occurs from the cytosol or after transit through the ER, we tagged a MOM-directed construct with a C-terminal N-glycosylation sequence. Although in vitro this construct was efficiently glycosylated by microsomes, the protein expressed in vivo localized almost exclusively to MOM, and was nearly completely unglycosylated. The small fraction of glycosylated protein was in the ER and was not a precursor to the unglycosylated form. Thus, targeting occurs directly from the cytosol. Moreover, ER and MOM compete for the same polypeptide, explaining the dual localization of some TA proteins.

Improving quality in emergency services to reduce hospital admission.

At the Emergency Department of Udine General Hospital (Italy) a programme to reduce admissions to the Internal Medicine Department was introduced in 1991. The majority of these admissions come from the Emergency Department, where many people, often without acute conditions, claim medical care. The programme consisted in organizational, professional and economic changes. At the end of 1991, the overall number of admissions to Udine General Hospital, as compared to 1990, decreased by 7.1%, but admissions to the Internal Medicine Department showed an 11.2% reduction. Finally, examinations for internal medical complaints in the emergency ward, not followed by hospitalization, increased by 15.5%. These results showed a reduction in admissions to the Internal Medicine Department greater than previously planned, with an increase in the number of outpatient examinations in the emergency room not followed by admission. Further targets were planned for 1992 to increase the quality of the service.

Psychotic and nonpsychotic bipolar mixed states: comparisons with manic and schizoaffective disorders.

We studied 259 female inpatients with bipolar spectrum disorders, of whom 26 (10%) were schizoaffective. Of the remaining 233, 61 (27%) met our conservatively set criteria for index mixed episodes (simultaneous presence of depressive and manic syndromes). These patients with rigorously defined mixed states were, in turn, about equally divided between psychotic and non-psychotic subgroups. The psychotic mixed patients (n = 32) were closer to the bipolar I pattern, and more often seem to come from a familial background of psychotic mood disorders; the nonpsychotic mixed patients (n = 29) conformed more closely to the bipolar II pattern, and more often had a hyperthymic and cyclothymic temperament and a family background of non-psychotic disorders and substance abuse. A three-way comparison between psychotic manic (n = 24), psychotic mixed (n = 32), and schizoaffective (n = 26) patients revealed few significant differences in temperamental, familial and course patterns. As expected, psychotic manic patients more often arose from a hyperthymic base and pursued a predominantly manic course; psychotic mixed patients were less likely to arise from such a base and more likely to pursue a mixed course. Finally, schizoaffective had earlier age at onset and longer duration of illness, suggesting that these patients had a more severe illness. Otherwise, interepisodic social adaptation was comparable in the three psychotic groups. The findings overall suggest that the presence of psychosis had relatively little impact on mixed states, which appear more based on temperamental characteristics.

Sensitive determination of free and plasma protein-bound dipyridamole by high-performance liquid chromatography.

For many years dipyridamole (DP) has been used in the treatment of hypertension as a vasodilator, but recently it has been recognised as an anti-platelet aggregation agent and to potentiate anti-metabolite activity. A rapid and sensitive (20 nM) procedure for the determination of free and protein-bound DP in plasma, using reversed-phase high-performance liquid chromatography on an Ultrasphere XL ODS (3 microns) column (70 mm x 4.6 mm I.D.) with ultraviolet detection (280 nm), is reported. Free and bound DP were separated using ultrafiltration. Concentrations of DP between 0.1 and 10 microM were measured in plasma with a relative standard deviation of less than 9.6%. The subsequent determination of DP levels in patients orally administered 450 mg per day showed that DP binding to plasma protein is higher than 90%.

Pseudo-Cushing's syndrome: an example of alcohol-induced central disorder in corticotropin-releasing factor-ACTH release?

Six chronic alcoholics with stigmata of Cushing's syndrome were studied before and after a period of alcohol abstinence. In all of them, after a minimum period of 3-4 weeks, a marked clinical and laboratory improvement was noted. The authors suggest that damage at brain level, with neurotransmitters' disturbance, caused by chronic alcoholism underlies the Pseudo-Cushing's Syndrome. The primum movens could be a disorder of the pituitary-adrenal axis secondary to a dysfunction of neurotransmitters with stimulation of ACTH-secreting cells of the adenohypophysis by the certicotropin-releasing factor (CRF).