Fat area and lipid droplet morphology of porcine oocytes during in vitro maturation with trans-10, cis-12 conjugated linoleic acid and forskolin

Lipid droplets (LD) in porcine oocytes form a dark mass reaching almost all cytoplasm. Herein we investigated changes in fat areas, cytoplasmic tone and LD morphology during in vitro maturation (IVM) of porcine oocytes cultured with 100mM trans-10, cis-12 conjugated linoleic acid (t10,c12 CLA) or 10mM forskolin at different time periods. Four groups were constituted: control, excipient, t10,c12 CLA and forskolin, with drugs being supplemented during 44 to 48h and the initial 22 to 24h in Experiments 1 and 2, respectively. In Experiment 3, forskolin was supplemented for the first 2 h. Matured oocytes were inseminated with frozen-thawed boar semen and cleavage rate recorded. Before and during IVM, samples of oocytes were evaluated for LD, total and fat areas and fat gray value or for meiotic progression. Results showed that forskolin supplementation during 44 to 48 h or 22 to 24 h inhibits oocyte maturation (exp. 1: forskolin = 5.1±8.0%, control = 72.6±5.0%; exp. 2: forskolin =24.3±7.4%, control =71.6±5.6%) and cleavage (exp. 1: forskolin=0.0±0.0%, control=55.4±4.1%; exp. 2: forskolin=8.3±3.3%, control=54.5±3.0%). Forskolin also reduced oocyte and fat areas. In Experiment 3, forskolin negative effect on oocyte maturation and cleavage disappeared, although minor (P<0.03) LD and oocyte fat areas were identified at 22 to 24 h of IVM. Oocytes supplemented with t10,c12 CLA during 44 to 48h presented a lighter (P<0.04) colour tone cytoplasm than those of control and forskolin. In conclusion, t10,c12 CLA and forskolin were capable of modifying the distribution and morphology of cytoplasmic LD during porcine oocyte maturation, thus reducing its lipid content in a time-dependent manner

Field and Molecular Epidemiology: How Viral Sequencing Changed Transmission Inferences in the First Portuguese SARS-CoV-2 Infection Cluster

Field epidemiology and viral sequencing provide a comprehensive characterization of transmission chains and allow a better identification of superspreading events. However, very few examples have been presented to date during the COVID-19 pandemic. We studied the first COVID-19 cluster detected in Portugal (59 individuals involved amongst extended family and work environments), following the return of four related individuals from work trips to Italy. The first patient to introduce the virus would be misidentified following the traditional field inquiry alone, as shown by the viral sequencing in isolates from 23 individuals. The results also pointed out family, and not work environment, as the primary mode of transmission.This work was funded by Portuguese Foundation for Science and Technology (FCT; Research4COVID19 projects 198_596862267, 617_613735895, 186_596855206). FCT also financed the PhD grant to NP (SFRH/BD/136299/2018) and post-doc grant to VF (SFRH/BPD/114927/2016)

Macroprolactinemia - laboratory determination and its clinical significance

Since prolactin (PRL) (a hormone produced by the anterior pituitary) was first identificated, the existence of hyperprolactinemic syndrome has been recognized. Main symptoms are galactorrhea, oligomenorrhea, amenorrhea and infertility in women and decreased libido and impotence in men. Macroprolactinemia reflects the predominance of circulating forms with reduced bioactivity not associated with typical clinical manifestations of hyperprolactinemia. It is identified by immunoassays commonly used in clinical practice, resulting in hyperprolactinemia. Polyetilenoglycol (PEG) is the most used method that removes PRL from serum. It is likely that the phenomenon of macroprolactinemia is consistently underestimated and unrecognized. Manufacturers of immunoassays for PRL have been slow to incorporate in the literature, validated protocols, and data related to the interference of PEG. From a clinical and biochemical point of view and, the main concern should be to avoid unnecessary investigation and treatment

The Percentage of [−2]Pro–Prostate-Specific Antigen and the Prostate Health Index Outperform Prostate-Specific Antigen and the Percentage of Free Prostate-Specific Antigen in the Detection of Clinically Significant Prostate Cancer and Can Be Used as Reflex Tests

Context.—: There is a need to avoid the overdiagnosis of prostate cancer (PCa) and to find more specific biomarkers. Objective.—: To evaluate the clinical utility of [-2]pro-prostate-specific antigen ([-2]proPSA) derivatives in detecting clinically significant PCa (csPCa) and to compare it with prostate-specific antigen (PSA) and with the percentage of free PSA (%fPSA). Design.—: Two hundred thirty-seven men (PSA: 2-10 ng/mL) scheduled for a prostate biopsy were enrolled. Parametric and nonparametric tests, receiver operating characteristic curves, and logistic regression analysis were applied. Outcomes were csPCa and overall PCa. Results.—: Both [-2]proPSA derivatives were significantly higher in csPCa and overall PCa (P < .001). The areas under the curves for the prediction of csPCa were higher for the percentage of [-2]proPSA (%[-2]proPSA) (0.781) and the prostate health index (PHI) (0.814) than for PSA (0.651) and %fPSA (0.724). There was a gain of 11% in diagnostic accuracy when %[-2]proPSA or PHI were added to a base model with PSA and %fPSA. Twenty-five percent to 29% of biopsies could have been spared with %[-2]proPSA (cutoff: ≥1.25%) and PHI (cutoff: ≥27), missing 10% of csPCas. The same results could have been achieved by using [-2]proPSA as a reflex test, when %fPSA was 25% or less (cutoffs: ≥1.12% and ≥24 for %[-2]proPSA and PHI, respectively). Conclusions.—: The [-2]proPSA derivatives improve the diagnostic accuracy of csPCa when the PSA value is between 2 and 10 ng/mL, sparing unnecessary biopsies and selecting patients for active surveillance. [-2]proPSA can be used as a reflex test when %fPSA is 25% or less, without reducing the diagnostic accuracy for csPCa and the number of spared biopsies.info:eu-repo/semantics/publishedVersio

Evaluation of urinary cysteinyl leukotrienes as biomarkers of severity and putative therapeutic targets in COVID-19 patients

Background Cysteinyl leukotrienes (CysLT) are potent inflammation-promoting mediators, but remain scarcely explored in COVID-19. We evaluated urinary CysLT (U-CysLT) relationship with disease severity and their usefulness for prognostication in hospitalized COVID-19 patients. The impact on U-CysLT of veno-venous extracorporeal membrane oxygenation (VV-ECMO) and of comorbidities such as hypertension and obesity was also assessed. Methods Blood and spot urine were collected in severe (n = 26), critically ill (n = 17) and critically ill on VV-ECMO (n = 17) patients with COVID-19 at days 1-2 (admission), 3-4, 5-8 and weekly thereafter, and in controls (n = 23) at a single time point. U-CysLT were measured by ELISA. Routine markers, prognostic scores and outcomes were also evaluated. Results U-CysLT did not differ between groups at admission, but significantly increased along hospitalization only in critical groups, being markedly higher in VV-ECMO patients, especially in hypertensives. U-CysLT values during the first week were positively associated with ICU and total hospital length of stay in critical groups and showed acceptable area under curve (AUC) for prediction of 30-day mortality (AUC: 0.734, p = 0.001) among all patients. Conclusions U-CysLT increase during hospitalization in critical COVID-19 patients, especially in hypertensives on VV-ECMO. U-CysLT association with severe outcomes suggests their usefulness for prognostication and as therapeutic targets.This work was supported by a RESEARCH 4 COVID-19 grant (project 519, reference number: 613690173) from FCT-Fundacao para a Ciencia e a Tecnologia (special support for rapid implementation projects for innovative response solutions to COVID-9 pandemic). CS-P is a recipient of a Ph.D. fellowship from FCT and MedInUP (UI/BD/150816/2020). P-PT was supported by a research contract within the scope of the RIFF-HEART project funded by FEDER via COMPETE, Portugal 2020-Operational Programme for Competitiveness and Internationalization (POCI) (POCI-01-0145-FEDER-032188) and by FCT (PTDC/MEC-CAR/32188/2017). Open access funding provided by FCT|FCCN (b-on)

Provenance of Cambrian–Ordovician Siliciclastic Rocks of Southwestern Iberia: Insights into the Evolution of the North Gondwana Margin

This study makes a comparison between the populations of detrital zircons of the Cambrian sandstones from the Ossa–Morena Zone (OMZ) and the Ordovician quartzites from the southern domains of the Central Iberian Zone (S-CIZ) to identify the sediment sources during the development of North Gondwana basins (southwestern Iberia). The U–Pb results obtained for the lower Cambrian sandstones of the OMZ show a remarkable similarity to the detrital zircon ages of greywackes from the underlying OMZ Ediacaran basement (the Série Negra succession). However, there is a greater proportion of Cryogenian grains in the Cambrian rocks, whose main sources are: (1) the late Cadomian magmatic arcs (Ediacaran, ca. 635–545 Ma) which also contributed to filling the late Ediacaran basins of the OMZ; and (2) the early Cadomian arcs (Cryogenian, ca. 700–635 Ma). In the Lower Ordovician quartzites of the S-CIZ (the Armorican and Sarnelha formations), the age distribution of detrital zircons overlaps the population of detrital zircons of the underlying S-CIZ Ediacaran basement (the Beiras Group). However, there are some differences in the Sarnelhas quartzites, which have a population of detrital zircons similar to those of the Ediacaran greywackes and Cambrian sandstones of the OMZ. The Cambrian grains found in the Lower Ordovician quartzites fit the ages of magmatism representing the onset of rifting in North Gondwana that is registered in the OMZ but absent from the S-CIZ. The early Ordovician zircon grains are probably related to the magmatic event that preceded the passive margin stage of the Rheic Ocean, and are found in both the CIZ and OMZ

Scale of Auditory Behaviors e testes auditivos comportamentais para avaliação do processamento auditivo em crianças falantes do português europeu

PURPOSE: The objective of this research was to assess the auditory abilities of Portuguese children and compare such abilities to the score of the Scale of Auditory Behaviors (SAB). METHODS: Fifty-one children were evaluated with audiometry, speech audiometry, acoustic immittance measures, and eight behavioral tests involving dichotic listening, monotic listening, temporal processing, and sound localization. Their parents filled in the SAB questionnaire adapted to European A. SAB scores and auditory tests scores were submitted to Pearson's correlation coefficient. RESULTS: There is significant correlation between the score on SAB questionnaire and the auditory processing tests. The greatest coefficient was observed in temporal processing test (p=0.000). CONCLUSION: There was correlation between the score of SAB and the performance in auditory processing tests, suggesting that the SAB may be used for auditory processing screening.OBJETIVO: Investigar as habilidades auditivas de crianças portuguesas e verificar se há correlação entre aquelas e o escore do Scale of Auditory Behaviors (SAB). MÉTODOS: Todas as crianças foram submetidas a audiometria tonal, logoaudiometria, medidas de imitância acústica e oito testes comportamentais do processamento auditivo, envolvendo tarefas de escuta dicótica, escuta monótica, processamento temporal e localização sonora. Os pais das 51 crianças portuguesas avaliadas preencheram o questionário SAB adaptado ao português europeu. Foram calculados os valores do coeficiente de correlação de Pearson entre os escores obtidos no questionário e os dos testes do processamento auditivo. RESULTADOS: Observou-se correlação significativa entre o escore do questionário e o dos testes comportamentais, tendo a maior sido observada nos testes relacionados ao processamento temporal (p=0,000). CONCLUSÃO: Houve correlação entre o escore da SAB e os resultados obtidos nos testes auditivos comportamentais em crianças portuguesas, sugerindo que este questionário pode ser utilizado em triagem do processamento auditivo.CIEC – Research Centre on Child Studies, UM (FCT R&D 317

Consolidated bioprocessing of corn cob-derived hemicellulose: engineered industrial Saccharomyces cerevisiae as efficient whole cell biocatalysts

Background Consolidated bioprocessing, which combines saccharolytic and fermentative abilities in a single microorganism, is receiving increased attention to decrease environmental and economic costs in lignocellulosic biorefineries. Nevertheless, the economic viability of lignocellulosic ethanol is also dependent of an efficient utilization of the hemicellulosic fraction, which contains xylose as a major component in concentrations that can reach up to 40% of the total biomass in hardwoods and agricultural residues. This major bottleneck is mainly due to the necessity of chemical/enzymatic treatments to hydrolyze hemicellulose into fermentable sugars and to the fact that xylose is not readily consumed by Saccharomyces cerevisiaethe most used organism for large-scale ethanol production. In this work, industrial S. cerevisiae strains, presenting robust traits such as thermotolerance and improved resistance to inhibitors, were evaluated as hosts for the cell-surface display of hemicellulolytic enzymes and optimized xylose assimilation, aiming at the development of whole-cell biocatalysts for consolidated bioprocessing of corn cob-derived hemicellulose. Results These modifications allowed the direct production of ethanol from non-detoxified hemicellulosic liquor obtained by hydrothermal pretreatment of corn cob, reaching an ethanol titer of 11.1 g/L corresponding to a yield of 0.328 g/g of potential xylose and glucose, without the need for external hydrolytic catalysts. Also, consolidated bioprocessing of pretreated corn cob was found to be more efficient for hemicellulosic ethanol production than simultaneous saccharification and fermentation with addition of commercial hemicellulases. Conclusions These results show the potential of industrial S. cerevisiae strains for the design of whole-cell biocatalysts and paves the way for the development of more efficient consolidated bioprocesses for lignocellulosic biomass valorization, further decreasing environmental and economic costs.This work has been carried out at the Biomass and Bioenergy Research Infrastructure (BBRI)-LISBOA-01-0145-FEDER-022059, supported by Operational Programme for Competitiveness and Internationalization (PORTUGAL2020), by Lisbon Portugal Regional Operational Programme (Lisboa 2020) and by North Portugal Regional Operational Programme (Norte 2020) under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and has been supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020, the “Contrato-Programa” UIDB/04050/2020, the MIT-Portugal Program (Ph.D. Grant PD/BD/128247/2016 to Joana T. Cunha) and through Project FatVal (POCI-01-0145-FEDER-032506) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio