Deletion of Tbc1d4/As160 abrogates cardiac glucose uptake and increases myocardial damage after ischemia/reperfusion.

BACKGROUND: Type 2 Diabetes mellitus (T2DM) is a major risk factor for cardiovascular disease and associated with poor outcome after myocardial infarction (MI). In T2DM, cardiac metabolic flexibility, i.e. the switch between carbohydrates and lipids as energy source, is disturbed. The RabGTPase-activating protein TBC1D4 represents a crucial regulator of insulin-stimulated glucose uptake in skeletal muscle by controlling glucose transporter GLUT4 translocation. A human loss-of-function mutation in TBC1D4 is associated with impaired glycemic control and elevated T2DM risk. The study's aim was to investigate TBC1D4 function in cardiac substrate metabolism and adaptation to MI. METHODS: Cardiac glucose metabolism of male Tbc1d4-deficient (D4KO) and wild type (WT) mice was characterized using in vivo [18F]-FDG PET imaging after glucose injection and ex vivo basal/insulin-stimulated [3H]-2-deoxyglucose uptake in left ventricular (LV) papillary muscle. Mice were subjected to cardiac ischemia/reperfusion (I/R). Heart structure and function were analyzed until 3 weeks post-MI using echocardiography, morphometric and ultrastructural analysis of heart sections, complemented by whole heart transcriptome and protein measurements. RESULTS: Tbc1d4-knockout abolished insulin-stimulated glucose uptake in ex vivo LV papillary muscle and in vivo cardiac glucose uptake after glucose injection, accompanied by a marked reduction of GLUT4. Basal cardiac glucose uptake and GLUT1 abundance were not changed compared to WT controls. D4KO mice showed mild impairments in glycemia but normal cardiac function. However, after I/R D4KO mice showed progressively increased LV endsystolic volume and substantially increased infarction area compared to WT controls. Cardiac transcriptome analysis revealed upregulation of the unfolded protein response via ATF4/eIF2α in D4KO mice at baseline. Transmission electron microscopy revealed largely increased extracellular matrix (ECM) area, in line with decreased cardiac expression of matrix metalloproteinases of D4KO mice. CONCLUSIONS: TBC1D4 is essential for insulin-stimulated cardiac glucose uptake and metabolic flexibility. Tbc1d4-deficiency results in elevated cardiac endoplasmic reticulum (ER)-stress response, increased deposition of ECM and aggravated cardiac damage following MI. Hence, impaired TBC1D4 signaling contributes to poor outcome after MI.

Adverse birth outcomes associated with Zika virus exposure during pregnancy in São José do Rio Preto, Brazil.

OBJECTIVES: We aimed to report the first 54 cases of pregnant women infected by Zika virus (ZIKV) and their virologic and clinical outcomes, as well as their newborns' outcomes, in 2016, after the emergence of ZIKV in dengue-endemic areas of São Paulo, Brazil. METHODS: This descriptive study was performed from February to October 2016 on 54 quantitative real-time PCR ZIKV-positive pregnant women identified by the public health authority of São José do Rio Preto, São Paulo, Brazil. The women were followed and had clinical and epidemiologic data collected before and after birth. Adverse outcomes in newborns were analysed and reported. Urine or blood samples from newborns were collected to identify ZIKV infection by reverse transcription PCR (RT-PCR). RESULTS: A total of 216 acute Zika-suspected pregnant women were identified, and 54 had the diagnosis confirmed by RT-PCR. None of the 54 women miscarried. Among the 54 newborns, 15 exhibited adverse outcomes at birth. The highest number of ZIKV infections occurred during the second and third trimesters. No cases of microcephaly were reported, though a broad clinical spectrum of outcomes, including lenticulostriate vasculopathy, subependymal cysts, and auditory and ophthalmologic disorders, were identified. ZIKV RNA was detected in 18 of 51 newborns tested and in eight of 15 newborns with adverse outcomes. CONCLUSIONS: Although other studies have associated many newborn outcomes to ZIKV infection during pregnancy, these same adverse outcomes were rare or nonexistent in this study. The clinical presentation the newborns we studied was mild compared to other reports, suggesting that there is significant heterogeneity in congenital Zika infection.

Influência da adubação e da cobertura morta na produtividade e no teor de flavonóides de Calendula officinalis L. (Asteraceae) / Influence of fertilization and mulching on the productivity and flavonoid content of Calendula officinalis L. (Asteraceae)

Desenvolveu-se ensaio de cultivo de Calendula officinalis, variando-se os tipos de adubação e utilização de cobertura morta na observação da produção de inflorescências, droga vegetal e quantificação dos flavonóides presentes nestes materiais.O delineamento experimental adotado foi de blocos casualizados com 3 repetições, em arranjo fatorial de 3 x 2, constituído de seis tratamentos, sendo 1. adubação química (100 kg de N ha-1 + 50 kg de P ha-1); 2. adubação química + cobertura morta (grama seca - Paspalum notatum); 3. adubação orgânica (esterco bovino curtido 50 t ha-1); 4. adubação orgânica + cobertura morta; 5. sem adubação química ou orgânica + cobertura morta; 6. sem adubação química ou orgânica e sem cobertura morta (testemunha). Os resultados de número capítulos produzidos, massa seca das inflorescências e massa da droga vegetal foram totalizados após cinco meses do plantio. A quantificação de flavonóides foi realizada através de método espectrofotométrico. Os dados foram analisados estatisticamente (ANOVA/Teste Tukey-Kramer-5 por cento) e mostraram diferenças significativas (70 por cento superior) no número de inflorescências, massa seca das inflorescências e massa da droga vegetal entre o tratamento com adubação orgânica (3) e os tratamentos sem adubação química ou orgânica com cobertura morta (5) e o testemunha (6). As demais comparações, inclusive o teor de flavonóides, entre os diversos tratamentos não resultaram em diferenças significativas.

Calendula officinalis was experimentally grown under field conditions with different fertilization types and mulching in order to evaluate inflorescence production, plant drugs and flavonoid levels. Experimental design was in randomized blocks, with 3 replicates, in a 3x2 factorial arrangement, consisting of six treatments, as follows: 1. chemical fertilization (100 kg N ha-1 + 50 kg P ha-1); 2. chemical fertilization + mulch (dry grass - Paspalum notatum); 3. organic fertilization (cured bovine manure 50 t ha-1); 4. organic fertilization + mulch; 5. no chemical or organic fertilization + mulch; 6. no chemical or organic fertilization and no mulch (control). Data regarding number of produced capitula, dry matter of inflorescences and plant drug matter were totaled after five months of planting. Flavonoids were quantified by spectrophotometry. The results were statistically analyzed (ANOVA / Tukey-Kramer test-5 percent) and showed significant differences (70 percent higher) for number of inflorescences, inflorescence dry matter and plant drug matter between the treatment with organic fertilization (3) and treatments with no chemical or organic fertilization with mulching (5) and control (6). The remaining comparisons, including flavonoid content, did not result in significant differences among treatments.