Heart Failure and Biomarkers. Unravelling new pathophysiological pathways, outcome predictors, and therapeutic targets.

The present thesis, result of the international experience I had during the last 3 years in the context of the International Philosophiæ Doctor programme in Cardiovascular Pathophysiology and Therapeutics (CardioPaTh), disserts on the known and unknown role of biomarkers in HF. The thesis is divided in several chapter, each of them dwelling upon specific aspects that have been investigated during the 3 years of my PhD curriculum. After a generalist paragraph dwelling upon the use of biomarkers in HF, its associated diseases, and the argument around the dilemma of racial differences (chapter 2), the important and emerging role of hormone deficiencies (HD), alone ore combined in the multiple hormone deficiency syndrome (MHDS) is discussed (chapter 3), together with the presentation of the results of the main project of my PhD, the T.O.S.CA. project, an international effort aimed to investigate the role of MHDS in HF. The following chapter (chapter 4) is based on the even strong relationship between the heart and the gut (the so-called Gut Axis), followed by a chapter showing the possible role of novel biomarkers in HF (chapter 5). In conclusion, a last chapter about my other HF research activity (not strictly related to biomarkers) is present

Klinefelter syndrome, insulin resistance, metabolic syndrome, and diabetes: review of literature and clinical perspectives

Klinefelter syndrome (KS), the most frequent chromosomic abnormality in males, is associated with hypergonadotropic hypogonadism and an increased risk of cardiovascular diseases (CVD). The mechanisms involved in increasing risk of cardiovascular morbidity and mortality are not completely understood. Insulin resistance, metabolic syndrome, and type 2 diabetes are more frequently diagnosed in KS than in the general population; however, the contribution of hypogonadism to metabolic derangement is highly controversial. Whether this dangerous combination of risk factors fully explains the CVD burden of KS patients remains unclear. In addition, testosterone replacement therapy only exerts a marginal action on the CVD system. This review summaries the current understandings of the complex relationship between KS, metabolic syndrome and cardiovascular risk in order to plan future studies and improve current strategies to reduce mortality in this high-risk population. Since fat accumulation and distribution seem to play a relevant role in triggering metabolic abnormalities, an early diagnosis and a tailored intervention strategy with drugs aimed at targeting excessive visceral fat deposition appear necessary in patients with KS

Growth hormone in heart failure revisited: An old story retold

Heart failure (HF) is a disease characterized by increasing prevalence, huge direct and indirect costs, and an ominous prognosis, worse than many cancers. At the beginning of the 90s, growth hormone (GH) was proposed as potential adjunctive therapy in HF mostly due to its growth-promoting, vasodilating, and anti-apoptotic actions. However, although several uncontrolled clinical studies showed that GH therapy improved several cardiovascular parameters, two robust trials failed to confirm these findings. Dwelling upon potential explanations for such apparent discrepancy led to the hypothesis that HF patients exhibit an inhomogeneous basal activity of the GH/insulin-like growth factor 1 (IGF-1) axis, ranging from GH/IGF-1 deficiency to GH resistance. This complex phenomenon was then reconsidered in the context of the so-called multiple hormone deficiency syndrome (MHD), that is the recognition that HF is characterized not only by the hyperactivation of several signaling pathways including the adrenergic, the renin-angiotensin-aldosterone and cytokine systems, but also by a reduced anabolic drive leading to a state of anabolic/catabolic imbalance. Mounting evidence support the concept that such imbalance is not a mere epiphenomen, since it exerts a significant impact on clinical performance and more importantly, on survival. Therefore, the paradigm shift to reconsider HF as MHD represented the underpinning to implement clinical trials focused on hormone replacement therapies in congestive heart failure (CHF). With regard to GH replacement therapy, one controlled single-blind study yielded promising results, and we are currently conducting a double-blind controlled trial, as well a large Registry study to evaluate the impact of MHD on HF progression

Progressive right ventricular dysfunction and exercise impairment in patients with heart failure and diabetes mellitus: insights from the T.O.S.CA. Registry

Findings from the T.O.S.CA. Registry recently reported that patients with concomitant chronic heart failure (CHF) and impairment of insulin axis (either insulin resistance-IR or diabetes mellitus-T2D) display increased morbidity and mortality. However, little information is available on the relative impact of IR and T2D on cardiac structure and function, cardiopulmonary performance, and their longitudinal changes in CHF

Effect of breeding techniques and prolonged post dry aging maturation process on biomolecule levels in raw buffalo meat

Recently, several concerns have been expressed on red meat quality and consumption. The aims of this study were to evaluate the influence of different breeding techniques and a prolonged post dry aging (PDA) maturation process on biomolecules level in raw buffalo meat. In the first experiment, two groups of animals were maintained with different space availability (15 vs. 10 m2/animal) for 90 days and biomolecules content was evaluated. In experiment 2, two diets (with or without ryegrass green forage) were used to assess the concentration of these biomolecules. Finally, in experiment 3, the meat of the animals that showed the highest content of biomolecules was chosen to assess the influence of the PDA maturation process. Buffaloes reared at 15 m2 showed a significantly (p < 0.05) higher content of the considered biomolecules compared with their counterparts. Similarly, buffaloes fed green forage showed higher content of biomolecules (p < 0.05) compared with the control group. The meat of the animals bred at 15 m2 and fed green forage showed a significant (p < 0.01) increase of biomolecules content during the PDA maturation process up to 60 days without influence microbiological profile in terms of total aerobic bacterial counts, yeasts, and molds. In conclusion, breeding techniques and PDA maturation system could enhance biomolecules levels in terms of quality, without affect health standards

The T.O.S.C.A. Project: Research, Education and Care

Despite recent and exponential improvements in diagnostic- therapeutic pathways, an existing “GAP” has been revealed between the “real world care” and the “optimal care” of patients with chronic heart failure (CHF). We present the T.O.S.CA. Project (Trattamento Ormonale dello Scompenso CArdiaco), an Italian multicenter initiative involving different health care professionals and services aiming to explore the CHF “metabolic pathophysiological model” and to improve the quality of care of HF patients through research and continuing medical education

Breed and Feeding System Impact the Bioactive Anti-Inflammatory Properties of Bovine Milk

In the present study, we aimed at assessing the influence of breed and feeding system on the bovine milk profile of betaines and carnitines and milk capacity in counteracting the inflammatory endothelial cell (EC) damage induced by interleukin (IL)-6. In the first experimental design, two breeds were chosen (Holstein vs. Modicana) to investigate the biomolecule content and antioxidant capacity in milk and dairy products. In the second experimental design, two feeding systems (pasture vs. total mixed ratio) were tested only in Holstein to evaluate the possible effect on the functional profile of milk and dairy products. Finally, the bulk milk from the two experimental designs was used to evaluate the efficacy of preventing IL-6-induced endothelial inflammatory damage. Results showed that Modicana milk and whey had higher biomolecule content and antioxidant activity compared to Holstein milk (p < 0.01). Milk from Holstein fed TMR showed higher concentration of γ-butyrobetaine, δ-valerobetaine (p < 0.01), and l-carnitine (p < 0.05). Similarly, whey from Holstein fed TMR also showed higher content of δ-valerobetaine, glycine betaine, l-carnitine, and acetyl-l-carnitine (p < 0.01) compared to the Holstein fed pasture. Conversely, the antioxidant activity of milk and dairy products was not affected by the feeding system. In ECs, all milk samples reduced the IL-6-induced cytokine release, as well as the accumulation of reactive oxygen species (ROS) and the induction of cell death, with the most robust effect elicited by Modicana milk (p < 0.01). Overall, Modicana milk showed a higher content of biomolecules and antioxidant activity compared to Holstein, suggesting that the breed, more than the feeding system, can positively affect the health-promoting profile of dairy cattle milk

Effects of the antioxidant crocin on frozen-thawed buffalo (Bubalus bubalis) sperm

This work aimed to evaluate the effect of crocin on frozen-thawed sperm quality in buffalo. Spermatozoa were incubated in Tyrode’s Albumin Lactate Pyruvate medium supplemented with 0, 0.5, 1, and 2 mM crocin for 2 h. Sperm motility was evaluated by phase-contrast microscopy, viability and acrosome integrity by Trypan blue Giemsa staining, and membrane functional integrity by the hypoosmotic swelling test. The DNA fragmentation was evaluated by Tunel and ROS levels by spectrofluorometric analysis. The treatment with 2 mM of crocin increased (P &lt;.05) sperm membrane functional integrity compared to the control group (59.1 ± 1.6 vs 53.3 ± 1.5) and reduced sperm DNA fragmentation, compared to the other groups (11.3 ± 1.1, 13.3 ± 1.2, 13.6 ± 1.2 and 6.0 ± 0.7, respectively in 0, 0.5, 1 and 2 mM crocin; P &lt;.01). Finally, a dose-dependent decrease (P &lt;.01) in superoxide anion production in the presence of crocin was observed, as indicated by Dihydroethidium values (922.6 ± 13.0, 596.8 ± 7.4, 498.9 ± 5.3 and 421.4 ± 5.0 a.u., respectively in 0, 0.5, 1 and 2 mM crocin; P &lt;.01). The results of this study demonstrated a positive effect of 2 mM crocin on frozen-thawed buffalo sperm, as indicated by the improvement of sperm membrane integrity and the reduction of DNA fragmentation and ROS levels.Highlights Crocin improves buffalo sperm quality. Crocin improves sperm membrane integrity and reduces DNA fragmentation. Crocin decreases oxidative stress in buffalo sperm

milk protein and cheese yield in buffalo species

Buffalo milk samples differing significantly for cheese yield values were analysed by 2D electrophoresis in order to outline a protein profile, with specific regards to k-casein fractions. Four buffaloes, two of which showing high cheese yield and two with low cheese yield selected from a group of 135 subjects were chosen for the proteomic analyses. Six main spots in 2D gels were recognized as αs1-, αs2-, β- and k-casein, α-lactoalbumin, β-lactoglobulin. The main visible differences in the 2D gels between buffaloes with high vs. low cheese yield were found in the appearance of the four k-casein spots (spots numbers:20, 19, 16, 18) which differ in the number of phosphorilation and glycosilation. The area and the intensity of the four spots were calculated by using Melanie II (Bio-Rad) software. Samples with high cheese yield showed higher value of the by-products: area x intensity of spot 16, correspondent to k-casein with one phosphorilation site, and lower values of spots 19 and 20, of k-casein with more than one phosphorilation site and glycosilated