NMR quantification of 16-O-methylcafestol and kahweol in Coffea canephora var. robusta beans from different geographical origins

Diterpenes have recently received a great deal of interest as tools to investigate the botanical origin of coffee. Specifically, kahweol has been proposed as a marker of Coffea arabica while 16-O-methylcafestol (16-OMC) is a Coffea canephora specific marker and its detection and quantification allow the authenticity of pure C. arabica roasted coffee blends to be assessed. In this study, we evaluated the possibility of the industrial use of the quantification of these diterpenes to assess the relative amounts of the two coffee species in blends. The content of 16-OMC and kahweol was determined in 78 samples (i.e., 39 green and the corresponding 39 roasted beans) of C. canephora from different geographical origins using a recently published NMR approach. Our results show a small natural variability in 16-OMC content for the Asian samples (average content = 1837 \ub1 113 mg/kg) while a much larger spread was found for the African samples (average content = 1744 \ub1 322 mg/kg). This large variability prevents the use of 16-OMC to quantify C. canephora in unknown roasted coffee blends. We also show that kahweol cannot be considered a specific C. arabica marker since it was detected almost all coffees and quantified in about 30% of the C. canephora samples

MR blockade protects against diet induced obesity, adipocyte dysfunction and cardiac inflammation in mice, through browning of the adipose organ and modulation of autophagy

Obesity is a key factor in the development of insulin resistance (IR), cardiovascular disease, hypertension, type 2 diabetes etc. Given the near epidemic incidence of obesity in western society there is a clear need for effective treatment options. Mineralocorticoid receptor (MR) blockade has shown significant promise in transgenic mouse models of obesity in limiting IR and adipocyte dysfunction, a disease that is independent of classical MR actions (renal). Female 10-weekold C57bl6 mice were fed with normal chow or a high fat (HF) diet for 12 weeks. Mice fed HF diet were concomitantly treated for 12 weeks with drospirenone (DRSP, 6 mg/kg/day), a potent MR antagonist with antiadipogenic activity, or spironolactone (SPIRO, 20 mg/kg/day). Mice fed HF diet showed a significant increase in total body weight, fat mass, mean adipocyte size, expression of white adipose tissue (WAT) marker genes and showed impaired glucose tolerance after intraperitoneal plasma glucose tolerance test. DRSP and SPIRO prevented weight gain and white fat mass expansion induced by HF diet in parametrial, perivescical, and inguinal depots without affecting interscapular fat pad weight. Magnetic Resonance Imaging (MRI) confirmed that MR antagonists blocked the HF dietdriven expansion of abdomino-pelvic (parametrial and perivescical) fat volume. High levels of MR mRNA were detected in all depots of adipose tissue. HF fed mice showed no increase in heart or kidney weight and tissue fibrosis. Cardiac macrophage recruitment and osteopontin staining was increased in hearts of HF fed mice and reversed by both MR antagonists. Moreover, both DRSP and SPIRO prevented the impaired glucose tolerance in mice fed HF diet, and countered HF diet-induced up-regulation of WAT markers transcripts and adipocyte hypertrophy. Importantly, MR antagonists increased uncoupling protein 1 (UCP-1) positive brown-like adipocyte content in WAT, and improved metabolic activity of adipose tissue, as indicated by PET/CT imaging. In keeping with this, MR antagonism significantly increased expression of brown-like adipocyte marker genes such PRDM16, CIDEA, beta-3 adrenergic receptor (ADRB3) and UCP-1 in all WAT depots analysed. In exploring the mechanism, we demonstrated that MR antagonism induced brown adipose tissue (BAT) markers, and reduced the autophagic rate, a key remodelling process in adipocyte differentiation, in WAT depots in vivo as well as in primary cultured adipocytes. We conclude that adipocyte MR regulates BAT-like remodeling of WAT through modulation of autophagy. MR blockade therefore has promise as a novel therapeutic option for the prevention of metabolic dysfunctions and the cardiac consequences of obesity. doi:10.1016/j.ijcme.2015.05.012 Transcriptional control of ICAM-1 in human coronary artery endothelial cells by Mineralocorticoid Receptor (MR): Implications for the protective effects of MR antagonists in cardiovascular diseases V. Marzolla, A. Armani, A. Fabbri, I.Z. Jaffe, M. Caprio Laboratory of Cardiovascular Endocrinology, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Raffaele Pisana, Rome, Italy Department of Medicina dei Sistemi, Endocrinology Unit, S. Eugenio & CTO A. Alesini Hospitals, University Tor Vergata, Rome, Italy Molecular Cardiology Research Institute, Tufts Medical Center, Boston

Androgens and Adipose Tissue in Males: A Complex and Reciprocal Interplay

Clinical evidence shows that in males obesity is frequently associated with hypogonadism and vice versa; also, low testosterone levels have been considered a “hallmark” of metabolic syndrome in men. These observations indicate that there is a strict connection between anatomically and functionally distinct cell types such as white adipocytes and Leydig cells, that synthesize testosterone. Adipose tissue is able to control several functions of the testis through its products secreted in the bloodstream. On the other hand, circulating levels of testosterone and estradiol deeply affect adipocyte proliferation, differentiation, and fat mass distribution, hereby controlling critical metabolic functions, such as food intake, insulin sensitivity, vascular reactivity, and immunity. This paper highlights the existing clinical and experimental evidence linking androgens and adipose tissue and illustrates the consequences occurring when the balance between fat mass distribution and eugonadism is lost

In vitro evaluation of sugar digestibility in molasses

Beet and cane molasses mainly contain mono- di-, and tri-saccharides, composed by hexoses, as well as pentoses in traces. However, rationing software consider sugars as only one entity, with a rate of digestion ∼20% h−1. The aim of this initial study was to investigate and evaluate the in vitro digestion dynamics and rates of the sugar fraction in molasses. Three beet and three cane molasses were randomly selected from a variety of samples collected world-wide and digested via in vitro rumen fermentation, at 1, 2, 3, 4, 6, 8, and 24 h. Samples were then analysed with a specific enzymatic kit to quantify residual sucrose, glucose, fructose, raffinose, galactose, and arabinose. Complete disappearance of sucrose happened within 3 hours of incubation. Glucose and fructose were completely digested within 4-6 h, showing variability among samples. Even if not so representative, galactose showed a similar trend of digestion (97% digestion within 3-4 h). Raffinose was quite slower in cane molasses, while it was completely digested within 1 h in beet molasses. Arabinose, a pentose, never reached a complete digestion, and its fermentation dynamic was different compared to other sugars. Calculated rates of digestion for sucrose, glucose and fructose, most representative sugars in molasses, were higher than 50% h−1 in both cane and beet. Obtained results showed that sugar fraction in molasses may vary, and different sugars are rapidly fermented by rumen microbes. Modern rationing models should consider a modification of sugar rates of digestion, since the actual one appears too slow than those observed in vitro.Highlights Molasses are unique blends of several sugars Major sugars are digested in few hours Rationing software should consider a faster rate of digestion for different sugars

Short communication: Characterization of molasses chemical composition

Beet and cane molasses are produced worldwide as a by-product of sugar extraction and are widely used in animal nutrition. Due to their composition, they are fed to ruminants as an energy source. However, molasses has not been properly characterized in the literature; its description has been limited to the type (sugarcane or beet) or to the amount of dry matter (DM), total or water-soluble sugars, crude protein, and ash. Our objective was to better characterize the composition of cane and beet molasses, examine possible differences, and obtain a proper definition of such feeds. For this purpose, 16 cane and 16 beet molasses samples were sourced worldwide and analyzed for chemical composition. The chemical analysis used in this trial characterized 97.4 and 98.3% of the compounds in the DM of cane and beet molasses, respectively. Cane molasses contained less DM compared with beet molasses (76.8 ± 1.02 vs. 78.3 ± 1.61%) as well as crude protein content (6.7 ± 1.8 vs. 13.5 ± 1.4% of DM), with a minimum value of 2.2% of DM in cane molasses and a maximum of 15.6% of DM in beet molasses. The amount of sucrose differed between beet and cane molasses (60.9 ± 4.4 vs. 48.8 ± 6.4% of DM), but variability was high even within cane molasses (39.2–67.3% of DM) and beet molasses. Glucose and fructose were detected in cane molasses (5.3 ± 2.7 and 8.1 ± 2.8% of DM, respectively), showing high variability. Organic acid composition differed as well. Lactic acid was more concentrated in cane molasses than in beet molasses (6.1 ± 2.8 vs. 4.5 ± 1.8% of DM), varying from 1.6 to 12.8% of DM in cane molasses. Dietary cation-anion difference showed numerical differences among cane and beet molasses (7 ± 53 vs. 66 ± 45 mEq/100 g of DM, on average). It varied from −76 to +155 mEq/100 g of DM in the cane group and from +0 to +162 mEq/100 g of DM in the beet group. Data obtained in this study detailed differences in composition between sources of molasses and suggested that a more complete characterization could improve the use of molasses in ration formulation

In vitro evaluation of sugar digestibility in molasses

Beet and cane molasses mainly contain mono- di-, and tri-saccharides, composed by hexoses, as well as pentoses in traces. However, rationing software consider sugars as only one entity, with a rate of digestion similar to 20% h(-1). The aim of this initial study was to investigate and evaluate the in vitro digestion dynamics and rates of the sugar fraction in molasses. Three beet and three cane molasses were randomly selected from a variety of samples collected world-wide and digested via in vitro rumen fermentation, at 1, 2, 3, 4, 6, 8, and 24 h. Samples were then analysed with a specific enzymatic kit to quantify residual sucrose, glucose, fructose, raffinose, galactose, and arabinose. Complete disappearance of sucrose happened within 3 hours of incubation. Glucose and fructose were completely digested within 4-6 h, showing variability among samples. Even if not so representative, galactose showed a similar trend of digestion (97% digestion within 3-4 h). Raffinose was quite slower in cane molasses, while it was completely digested within 1 h in beet molasses. Arabinose, a pentose, never reached a complete digestion, and its fermentation dynamic was different compared to other sugars. Calculated rates of digestion for sucrose, glucose and fructose, most representative sugars in molasses, were higher than 50% h(-1) in both cane and beet. Obtained results showed that sugar fraction in molasses may vary, and different sugars are rapidly fermented by rumen microbes. Modern rationing models should consider a modification of sugar rates of digestion, since the actual one appears too slow than those observed in vitro

The role of dose size in a chemotherapy regimen (ProMECE-CytaBOM) for the first-line treatment of large B-cell lymphomas: a randomized trial by the Gruppo Italiano Studio Linfomi (GISL)

Background: It is still unclear the actual contribute of dose intensity (DI), dose size (DS) and dose density (DD) in the conventional chemotherapy of large, B-cell non-Hodgkin lymphomas. Methods: A prospective, randomized trial compared the cyclic schedule of ProMECE-CytaBOM chemotherapy (cyc-PC, 6 cycles) with a modified version of it, which administered the same drugs sequentially (seq-PC), with the same planned cumulative DI and an 83% DD, within the same time frame (113 days), but with three times higher DS of all the drugs except vincristine. Results: Fifty-six patients received cyc-PC and 52 seq-PC. The actual mean cumulative DI was 0.79 +/- 0.15 with cyc-PC, 0.78 +/- 0.17 with seq-PC. Response was complete in 59% and 52%, partial in 20% and 21%, null in 5% and 6%, respectively. There were four toxic deaths (two per arm). Relapses occurred in 36% and 37%, respectively. Toxicity was similar in both arms. Overall, failure-free, progression-free and disease-free survival (median follow-up: 54 months) were statistically indifferent. Conclusions: The very similar DI actually delivered in both arm seems to be the main common determinant of the indifferent results recorded. Increasing DS - at least within the limits clinically attainable without stem cell rescue - does not improve results

Essential role of ICAM-1 in aldosterone-induced atherosclerosis.

OBJECTIVE: Elevated aldosterone is associated with increased risk of atherosclerosis complications, whereas treatment with mineralocorticoid receptor (MR) antagonists decreases the rate of cardiovascular events. Here we test the hypothesis that aldosterone promotes early atherosclerosis by modulating intercellular adhesion molecule-1 (ICAM-1) expression and investigate the molecular mechanisms by which aldosterone regulates ICAM-1 expression. METHODS AND RESULTS: Apolipoprotein-E (ApoE)-/- mice fed an atherogenic diet and treated with aldosterone for 4weeks showed increased vascular expression of ICAM-1, paralleled by enhanced atherosclerotic plaque size in the aortic root. Moreover, aldosterone treatment resulted in increased plaque lipid and inflammatory cell content, consistent with an unstable plaque phenotype. ApoE/ICAM-1 double knockout (ApoE-/-/ICAM-1-/-) littermates were protected from the aldosterone-induced increase in plaque size, lipid content and macrophage infiltration. Since aldosterone is known to regulate ICAM-1 transcription via MR in human endothelial cells, we explored MR regulation of the ICAM-1 promoter. Luciferase reporter assays performed in HUVECs using deletion constructs of the human ICAM-1 gene promoter showed that a region containing a predicted MR-responsive element (MRE) is required for MR-dependent transcriptional regulation of ICAM-1. CONCLUSIONS: Pro-atherogenic effects of aldosterone are mediated by increased ICAM-1 expression, through transcriptional regulation by endothelial MR. These data enhance our understanding of the molecular mechanism by which MR activation promotes atherosclerosis complications

Evaluation of Brix Refractometry to Estimate Immunoglobulin G Content in Buffalo Colostrum and Neonatal Calf Serum

Brix refractometry has been widely demonstrated to be a useful tool for monitoring colostrum management program and passive immunity transfer (PIT) in Bovines, but its suitability has never been verified in Buffalo. Therefore, the objective of this study was to evaluate the utility of a simple and rapid tool such as a digital Brix refractometer to estimate colostrum quality and for evaluating the success of passive transfer of immunoglobulin G (IgG) in Buffalo calves. The optimal cut points levels for Brix Refractometry for distinguishing good- and poor-quality colostrum and for assessing the adequacy of passive immunity transfer in calves were determined. For this aim, 26 first-milking maternal colostrum (MC) were collected from first-calf heifers. Blood samples were obtained from their calves at birth (T0) and 72 hours after (T3). Colostrum and Serum IgG content were determined by indirect enzyme-linked immunosorbent assay (ELISA), whereas total protein (TP, g/dL) and percentage Brix (%Brix) by means of a digital Brix refractometer. The mean colostrum IgG was 64.9 ± 29.3 mg/mL. The mean serum %Brix at T3 was 9.6 ± 0.9%. The mean serum IgG content at T3 was 11.1 ± 2.0 mg/mL. Pearson’s correlation coefficient (rp) was determined between Brix and ELISA measurements: colostrum %Brix showed a significant correlation with serum %Brix (rp = 0.82, p < 0.001); serum %Brix was highly correlated with serum TP (STP, g/dL) (rp = 0.98, p < 0.001) and serum IgG (mg/mL) (rp = 0.85, p < 0.001). A cut point of 18% Brix to estimate samples of MC ≥ 50 mg/mL from first-calf heifers was more appropriate for the buffalo. A cut point of 8.4% Brix resulted in the greatest percentage of calf serum samples being correctly classified. Based on our findings, a digital Brix refractometer could be a useful tool to monitor colostrum quality and to estimate PIT in Buffalo calves

Calmodulin enhances cryptochrome binding to INAD in Drosophila photoreceptors

Light is the main environmental stimulus that synchronizes the endogenous timekeeping systems in most terrestrial organisms. Drosophila cryptochrome (dCRY) is a light-responsive flavoprotein that detects changes in light intensity and wavelength around dawn and dusk. We have previously shown that dCRY acts through Inactivation No Afterpotential D (INAD) in a light-dependent manner on the Signalplex, a multiprotein complex that includes visual-signaling molecules, suggesting a role for dCRY in fly vision. Here, we predict and demonstrate a novel Ca2+-dependent interaction between dCRY and calmodulin (CaM). Through yeast two hybrid, coimmunoprecipitation (Co-IP), nuclear magnetic resonance (NMR) and calorimetric analyses we were able to identify and characterize a CaM binding motif in the dCRY C-terminus. Similarly, we also detailed the CaM binding site of the scaffold protein INAD and demonstrated that CaM bridges dCRY and INAD to form a ternary complex in vivo. Our results suggest a process whereby a rapid dCRY light response stimulates an interaction with INAD, which can be further consolidated by a novel mechanism regulated by CaM