Effects of ranolazine on astrocytes and neurons in primary culture

Ranolazine (Rn) is an antianginal agent used for the treatment of chronic angina pectoris when angina is not adequately controlled by other drugs. Rn also acts in the central nervous system and it has been proposed for the treatment of pain and epileptic disorders. Under the hypothesis that ranolazine could act as a neuroprotective drug, we studied its effects on astrocytes and neurons in primary culture. We incubated rat astrocytes and neurons in primary cultures for 24 hours with Rn (10−7, 10−6 and 10−5 M). Cell viability and proliferation were measured using trypan blue exclusion assay, MTT conversion assay and LDH release assay. Apoptosis was determined by Caspase 3 activity assay. The effects of Rn on proinflammatory mediators IL-β and TNF-α was determined by ELISA technique, and protein expression levels of Smac/Diablo, PPAR-γ, Mn-SOD and Cu/Zn-SOD by western blot technique. In cultured astrocytes, Rn significantly increased cell viability and proliferation at any concentration tested, and decreased LDH leakage, Smac/Diablo expression and Caspase 3 activity indicating less cell death. Rn also increased anti-inflammatory PPAR-γ protein expression and reduced pro-inflammatory proteins IL-1 β and TNFα levels. Furthermore, antioxidant proteins Cu/Zn-SOD and Mn-SOD significantly increased after Rn addition in cultured astrocytes. Conversely, Rn did not exert any effect on cultured neurons. In conclusion, Rn could act as a neuroprotective drug in the central nervous system by promoting astrocyte viability, preventing necrosis and apoptosis, inhibiting inflammatory phenomena and inducing anti-inflammatory and antioxidant agents

RICORS2040 : The need for collaborative research in chronic kidney disease

Chronic kidney disease (CKD) is a silent and poorly known killer. The current concept of CKD is relatively young and uptake by the public, physicians and health authorities is not widespread. Physicians still confuse CKD with chronic kidney insufficiency or failure. For the wider public and health authorities, CKD evokes kidney replacement therapy (KRT). In Spain, the prevalence of KRT is 0.13%. Thus health authorities may consider CKD a non-issue: very few persons eventually need KRT and, for those in whom kidneys fail, the problem is 'solved' by dialysis or kidney transplantation. However, KRT is the tip of the iceberg in the burden of CKD. The main burden of CKD is accelerated ageing and premature death. The cut-off points for kidney function and kidney damage indexes that define CKD also mark an increased risk for all-cause premature death. CKD is the most prevalent risk factor for lethal coronavirus disease 2019 (COVID-19) and the factor that most increases the risk of death in COVID-19, after old age. Men and women undergoing KRT still have an annual mortality that is 10- to 100-fold higher than similar-age peers, and life expectancy is shortened by ~40 years for young persons on dialysis and by 15 years for young persons with a functioning kidney graft. CKD is expected to become the fifth greatest global cause of death by 2040 and the second greatest cause of death in Spain before the end of the century, a time when one in four Spaniards will have CKD. However, by 2022, CKD will become the only top-15 global predicted cause of death that is not supported by a dedicated well-funded Centres for Biomedical Research (CIBER) network structure in Spain. Realizing the underestimation of the CKD burden of disease by health authorities, the Decade of the Kidney initiative for 2020-2030 was launched by the American Association of Kidney Patients and the European Kidney Health Alliance. Leading Spanish kidney researchers grouped in the kidney collaborative research network Red de Investigación Renal have now applied for the Redes de Investigación Cooperativa Orientadas a Resultados en Salud (RICORS) call for collaborative research in Spain with the support of the Spanish Society of Nephrology, Federación Nacional de Asociaciones para la Lucha Contra las Enfermedades del Riñón and ONT: RICORS2040 aims to prevent the dire predictions for the global 2040 burden of CKD from becoming true

Dietary α‐Linolenic Acid, Marine ω‐3 Fatty Acids, and Mortality in a Population With High Fish Consumption: Findings From the PREvención con DIeta MEDiterránea (PREDIMED) Study

Background: Epidemiological evidence suggests a cardioprotective role of α‐linolenic acid (ALA), a plant‐derived ω‐3 fatty acid. It is unclear whether ALA is beneficial in a background of high marine ω‐3 fatty acids (long‐chain n‐3 polyunsaturated fatty acids) intake. In persons at high cardiovascular risk from Spain, a country in which fish consumption is customarily high, we investigated whether meeting the International Society for the Study of Fatty Acids and Lipids recommendation for dietary ALA (0.7% of total energy) at baseline was related to all‐cause and cardiovascular disease mortality. We also examined the effect of meeting the society's recommendation for long‐chain n‐3 polyunsaturated fatty acids (≥500 mg/day). Methods and Results: We longitudinally evaluated 7202 participants in the PREvención con DIeta MEDiterránea (PREDIMED) trial. Multivariable‐adjusted Cox regression models were fitted to estimate hazard ratios. ALA intake correlated to walnut consumption (r=0.94). During a 5.9‐y follow‐up, 431 deaths occurred (104 cardiovascular disease, 55 coronary heart disease, 32 sudden cardiac death, 25 stroke). The hazard ratios for meeting ALA recommendation (n=1615, 22.4%) were 0.72 (95% CI 0.56–0.92) for all‐cause mortality and 0.95 (95% CI 0.58–1.57) for fatal cardiovascular disease. The hazard ratios for meeting the recommendation for long‐chain n‐3 polyunsaturated fatty acids (n=5452, 75.7%) were 0.84 (95% CI 0.67–1.05) for all‐cause mortality, 0.61 (95% CI 0.39–0.96) for fatal cardiovascular disease, 0.54 (95% CI 0.29–0.99) for fatal coronary heart disease, and 0.49 (95% CI 0.22–1.01) for sudden cardiac death. The highest reduction in all‐cause mortality occurred in participants meeting both recommendations (hazard ratio 0.63 [95% CI 0.45–0.87]). Conclusions: In participants without prior cardiovascular disease and high fish consumption, dietary ALA, supplied mainly by walnuts and olive oil, relates inversely to all‐cause mortality, whereas protection from cardiac mortality is limited to fish‐derived long‐chain n‐3 polyunsaturated fatty acids. Clinical Trial Registration URL: http://www.Controlled-trials.com/. Unique identifier: ISRCTN35739639

Anthropometric analysis and performance characteristics to predict selection in young male and female handball players

Abstract The aim of this study was two-fold. The first aim was to determine if there were any anthropometric and physical performance differences (controlling for maturation) between male and female handball players selected in training categories as well asthe relation of these differences with the performance level achieved. The second aim was to identify the discriminatory variables between the performance levels achieved. A total of 216 young handball players (125 men and 91 women) participated in the study. The data were classified by selection level (regional n=154; national n=62), gender (men; women) and age category (under-15; under-17). The use of MANCOVA analyses, controllingfor maturation, identified how gender could determine variables related to handball players' future competitive levels. The results revealed that anthropometric variables such as height, arm span, trochanter height, thigh girth, and leg girth were more influential in men than in women. In addition, the physical performance tests of vertical jump (squat jump and counter movement jump with/without arm) and 10x5m shuttle run were determinants in both sexes. Discriminatory analysis predicted that a combination of five variables (counter movement jump with arm, body mass, 10x5m shuttle run, dominant hand length and trochanter height) would successfully distinguish between regional and national players, with a predictive accuracy of 81.9% for all players

Cell viability was determined by MTT assay in cells treated for 24 h.

<p>Astrocytes were incubated without Rn (control, C), with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M), with Amyloid β_1–42 (15 μM) or with Amyloid β_1–42 (15 μM) + Rn (10⁻⁶ M) (panel A). Neurons were incubated without (control, C) or with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M) (panel B). Data are mean ± SD of four independent experiments (four different rats). *<i>p</i> < 0.05 <i>vs</i>. control.</p

Smac/Diablo protein expression.

<p>Astrocytes (panel A) or neurons (panel B) were incubated without Rn (control, C) or with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M) for 24 h and collected to determine Smac/Diablo protein expression by Western blot. A representative immunoblot is shown in the top panel. Data are mean ± SD of four independent experiments (four different rats). *<i>p</i> < 0.05 vs. control.</p

Mn-SOD protein expression.

<p>Astrocytes (panel A) or neurons (panel B) were incubated without Rn (control, C) or with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M) for 24 h and collected to determine Mn-SOD protein expression by Western blot. A representative immunoblot is shown in the top panel. Data are mean ± SD of four independent experiments (four different rats). *p < 0.05 vs. control.</p

Cytokine TNF-α determination.

<p>Astrocytes (panel A) or neurons (panel B) were incubated without Rn (control, C) or with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M) and cell culture supernatants were harvested. TNF-α secretion were determined by ELISA. Values are means ± SD of replicate experiments from four independent cell experiments (four different rats). *<i>p</i> < 0.05 <i>vs</i> control.</p

PPAR-γ protein expression.

<p>Astrocytes (panel A) or neurons (panel B) were incubated without Rn (control, C) or with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M) for 24 h and collected to determine PPAR-γ protein expression by Western blot. A representative immunoblot is shown in the top panel. Data are mean ± SD of four independent experiments (four different rats). *<i>p</i> < 0.05 vs. control.</p

Cytokine IL-1β determination.

<p>Astrocytes (panel A) or neurons (panel B) were incubated without Rn (control, C) or with Rn (10⁻⁷, 10⁻⁶, 10⁻⁵ M) and cell culture supernatants were harvested. IL-1β secretion were determined by ELISA. Values are means ± SD of replicate experiments from four independent experiments (four different rats). *<i>p</i> < 0.05 <i>vs</i> control.</p