-We examined the effects of diet composition and fasting on lipolysis of freshly isolated adipocytes from gilthead seabream (Sparus aurata). We also analyzed the effects of insulin, glucagon, and growth hormone (GH) in adipocytes isolated from fish fed with different diets. Basal lipolysis, measured as glycerol release, increased proportionally with cell concentration and time of incubation, which validates the suitability of these cell preparations for the study of hormonal regulation of this metabolic process. Gilthead seabream were fed two different diets, FM (100% of fish meal) and PP (100% of plant protein supplied by plant sources) for 6 wk. After this period, each diet group was divided into two groups: fed and fasted (for 11 days). Lipolysis was significantly higher in adipocytes from PP-fed fish than in adipocytes from FM-fed fish. Fasting provoked a significant increase in the lipolytic rate, about threefold in isolated adipocytes regardless of nutritional history. Hormone effects were similar in the different groups: glucagon increased the lipolytic rate, whereas insulin had almost no effect. GH was clearly lipolytic, although the relative increase in glycerol over control was lower in isolated adipocytes from fasted fish compared with fed fish. Together, we demonstrate for the first time that lipolysis, measured in isolated seabream adipocytes, is affected by the nutritional state of the fish. Furthermore, our data suggest that glucagon and especially GH play a major role in the control of adipocyte lipolysis.fish; nutritional and hormonal regulation; insulin; glucagon; growth hormone; fasting ADIPOSE TISSUE PLAYS A CENTRAL role in energy homeostasis in storing lipids in the form of triacylglycerols and in mobilizing them via breakdown into free fatty acids (FFAs) and glycerol (34). Adipose tissue is one of the most important lipid stores in several teleosts, although in some species liver or muscle also constitutes lipid storage organs (33). In salmonids, adipose tissue is distributed primarily in the abdominal cavity, associated with the mesenteric and pyloric ceca (31). In gilthead seabream, adipose tissue is also located periviscerally. It is known that fish adiposity changes seasonally and is affected by trophic status. High-fat feeds can lead to increases in visceral fat (4), resulting in reduced product yield and quality of cultured fish (7). In gilthead seabream, replacing fish meal with plant protein seems to alter lipid metabolism and results in smaller fat depots (10).Endocrine control of adipose tissue mobilization and storage remains almost unexplored in fish, although insulin and glucagon are clearly involved (12,24,28). Key hepatic enzymes in lipid metabolism such as hepatic lipase and acetyl-CoA carboxylase are also regulated by pancreatic hormones in vitro in isolated hepatocytes or in vivo in injected fish (13,21). Several in vivo studies suggest that growth hormone (GH) and somatolactin act together, in a complementary way, to regulate fat stores in gilthead seabream (4, 20). Howev...

This report presents our experience with T therapy in a cohort of T-deficient men on active surveillance (AS) for Gleason 3 + 3 and Gleason 3 + 4 prostate cancer (PCa). A retrospective chart review identified 28 men with T deficiency who underwent T therapy (T group) for at least 6 months while on AS for PCa. A comparison group of 96 men on AS for PCa with untreated T deficiency (no-T group) was identified at the same institution. The AS protocol followed a modified Epstein criteria and allowed inclusion of men with a single core of low-volume Gleason 3 + 4 PCa. Mean age was 59.5 and 61.3 years, and mean follow-up was 38.9 and 42.4 months for the T and no-T groups, respectively. Of all 28 men in the T group, 3 (10.7%) men developed an increase in Gleason score while on AS. Of 22 men in the T group with Gleason 3 + 3 disease, 7 (31.8%) men developed biopsy progression including 3 men (13.6%) who developed Gleason 3 + 4 PCa. Of 6 men with Gleason 3 + 4 disease at baseline, 2 (33.3%) men developed an increase in tumor volume, and none developed upgrading beyond Gleason 3 + 4. All 96 men in the no-T group had Gleason 3 + 3 disease at baseline and, 43 (44.7%) developed biopsy progression, including 9 men (9.38%) with upgrading to Gleason 7 (3 + 4). Biopsy progression rates were similar for both groups and historical controls. Biopsy progression in men on AS appears unaffected by T therapy over 3 years. Prospective placebo-controlled trials of T therapy in T-deficient men on AS should be considered given the symptomatic benefits experienced by treated men.

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Most of the ongoing projects aimed at the development of specific therapies and vaccines against COVID-19 use the SARS-CoV-2 spike (S) protein as the main target. The binding of the spike protein with the ACE2 receptor (ACE2) of the host cell constitutes the first and key step for virus entry. During this process, the receptor binding domain (RBD) of the S protein plays an essential role, since it contains the receptor binding motif (RBM), responsible for the docking to the receptor. So far, mostly biochemical methods are being tested in order to prevent binding of the virus to ACE2. Here we show, with the help of atomistic simulations, that external electric fields of easily achievable and moderate strengths can dramatically destabilise the S protein, inducing long-lasting structural damage. One striking field-induced conformational change occurs at the level of the recognition loop L3 of the RBD where two parallel beta sheets, believed to be responsible for a high affinity to ACE2, undergo a change into an unstructured coil, which exhibits almost no binding possibilities to the ACE2 receptor. We also show that these severe structural changes upon electric-field application also occur in the mutant RBDs corresponding to the variants of concern (VOC) B.1.1.7 (UK), B.1.351 (South Africa) and P.1 (Brazil). Remarkably, while the structural flexibility of S allows the virus to improve its probability of entering the cell, it is also the origin of the surprising vulnerability of S upon application of electric fields of strengths at least two orders of magnitude smaller than those required for damaging most proteins. Our findings suggest the existence of a clean physical method to weaken the SARS-CoV-2 virus without further biochemical processing. Moreover, the effect could be used for infection prevention purposes and also to develop technologies for in-vitro structural manipulation of S. Since the method is largely unspecific, it can be suitable for application to other mutations in S, to other proteins of SARS-CoV-2 and in general to membrane proteins of other virus types.

ObjectiveTo determine whether total testosterone and free testosterone levels predict disease reclassification in a cohort of men with prostate cancer (PCa) on active surveillance (AS). Patients and MethodsTotal testosterone and free testosterone concentrations were determined at the time the men began the AS protocol. Statistical analysis was performed using Student's t-test and a chi-squared test to compare groups. Odds ratios (ORs) with 95% confidence intervals (CIs) were obtained using univariate logistic regression. Receiver-operator characteristic curves were generated to determine the investigated testosterone thresholds. Kaplan-Meier curves were used to estimate time to disease reclassification. A Cox proportional hazard regression model was used for multivariate analysis. ResultsA total of 154 men were included in the AS cohort, of whom 54 (35%) progressed to active treatment. Men who had disease reclassification had significantly lower free testosterone levels than those who were not reclassified (0.75 vs 1.02 ng/dL, P = 0.03). Men with free testosterone levels <0.45 ng/dL had a higher rate of disease reclassification than patients with free testosterone levels ≥0.45 (P = 0.032). Free testosterone levels <0.45 ng/dL were associated with a several-fold increase in the risk of disease reclassification (OR 4.3,). Multivariate analysis showed that free testosterone and family history of PCa were independent predictors of disease reclassification. ConclusionsFree testosterone levels were lower in men with PCa who had reclassification during AS. Men with moderately severe reductions in free testosterone level are at increased risk of disease reclassification.

BackgroundThe objective of this work was to assess the overall survival, cause-specific survival and biochemical failure-free survival of a contemporary cohort of patients with localized prostate cancer (PCa) treated with intensity-modulated radiation therapy (IMRT) or radical prostatectomy (RP).MethodsWe did a retrospective cohort study of our institution’s registry of patients undergoing either IMRT or RP between January 1999 and March 2010, and assessed Prostate Specific Antigen (PSA), age at diagnosis, Gleason score, and digital rectal examination. Two groups were separated according to RP or IMRT treatment and these groups were in turn divided into risk groups according to the D’Amico classification. Overall survival (OS), cause-specific survival (CSS), mortality from other causes (MOC), and biochemical disease-free survival (BDFS) were assessed.ResultsTwelve-hundred patients were included: 993 in the RP group and 207 in the IMRT group.The IMRT group had older age, PSA at diagnosis and a significantly higher percentage of cancer on the needle biopsy (p <0.001). Of the 207 patients who underwent IMRT, 54% presented comorbidities. Median follow-up was 91.7 months for the RP group and 76 months for the IMRT group. The OS at 5 and 7 was 96.2, and 93.7 for the RP group respectively and 88.4, and 83.1 for the IMRT group respectively (p <0.001). There were no significant differences in the CSS in relation to treatment received among the low- and high-risk groups, while in the intermediate-risk group, patients who underwent to RP had a higher CSS than patients who underwent IMRT (99.6% vs 94.1%, p = 0.003). The IMRT group had a significantly better BDFS than the RP group (86.4% vs. 74.3%, respectively, p = 0.016).ConclusionsPatients treated with RP were significantly younger and had a better prognosis than patients treated using IMRT, and according to our results, RP had better outcomes in terms of OS while IMRT had greater MOC. Treatment modality did not affect the CSS.

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