Growth, pectoralis muscle performance, and testis of pelung cockerels (Gallus gallus gallus [Linnaeus, 1758]) supplemented with blood clam shell powder (Anadara granosa [Linnaeus, 1758]).

Background and Aim: Pelung cockerels (Gallus gallus gallus [Linnaeus, 1758]) are different from other native cockerels in that they have a long and unique voice, in addition to their tall, large, and sturdy body with a relatively heavy body weight (BW). The sound quality of pelung cockerels is affected by the structure of the syrinx and their large and strong chest muscles. The performance of the chest muscles, and subsequently its voice, is influenced by the hormone testosterone. The shell of blood clams (Anadara granosa Linnaeus, 1758), a saltwater bivalve is known to contain a natural aromatase blocker (NAB) capable of blocking the aromatase enzyme from converting testosterone to estradiol. This generates consistently high levels of testosterone. This study aimed to determine the effect of blood clam shell powder (BCSP) as an NAB on the growth, pectoralis muscle performance, and testes of pelung cockerels. Materials and Methods: The study design was a completely randomized design, with 16 pelung cockerels aged 40-56 weeks divided into four treatment groups: T0 (control); T1 (BCSP [A. granosa] 0.9 mg/kg BW); T2 (zinc sulfate [ZnSO4] 0.9 mg/kg BW); and T3 (testosterone 3 mg/day). The animals were acclimatized for 7 days and then given dietary treatments for 56 days. The measurement of the comb, wattle, and chest circumference (CC) of pelung cockerels was performed on days 0, 14, 28, 42, and 56. At the end of the treatment, the pelung cockerels were sacrificed and the data of the pectoralis muscle weight (PMW), testis weight (TW), and area of the pectoralis muscle (APM) were measured. Samples of pectoralis muscle and testes were taken and fixed in 10% neutral buffer formalin for histology. The proliferating cell nuclear antigen (PCNA) was identified by immunohistochemical staining. To measure fascicle area (FA), myofiber area (MA), and enumerate, the fascicle myofibers (NM) histology preparations were stained with hematoxylin and eosin (H and E). Testicular preparations were stained with H and E to measure the diameter of the seminiferous tubules (DST) using ImageJ software. Results: The growth performance on day 56 showed significantly (p < 0.05) higher differences of CC in T1 compared to T2 and T0, in T1 and T3 compared to T0, and in T3 and T2 compared to T0. Pectoralis muscle results, that is, FA, NM, MA, and PCNA-positive cells, showed that cockerels on treatment T3 had significantly higher results than other treatments, T1 was significantly different from T2 and T0, and T2 was significantly different from T0. In addition, the TW and DST measurement of cockerels on treatment T3 were significantly reduced (p < 0.05) than the other treatment groups. Conclusion: The oral administration of BCSP in the role of a NAB at a dose of 0.9 mg/kg BW for 56 days improved the growth performance and pectoralis muscle, especially the CC, FA, NM, MA, and PCNA-positive cells parameters, but did not affect the PMW, APM, and testis of pelung cockerels. The administration of testosterone at 3 mg/day for 56 days contributed to the decrease in TW and DST, as well as atrophy of the seminiferous tubules of pelung cockerels.

Anadara granosa shell powder improves the metabolism, testosterone level, and sound frequency of Pelung chickens.

BACKGROUND AND AIM: Pelung chickens make unique, pleasant, and rhythmic sounds in addition to having strong muscle mass. Performance is controlled by testosterone. A natural aromatase blocker is an agent capable of blocking the aromatase enzyme, which consequently prevents testosterone from being changed into estradiol. Such a condition results in consistently high testosterone levels. The aim of this study was to determine the effect of the administration of the shell powder of Anadara granosa on the testosterone level, frequency and quality of sounds, and metabolism of Pelung chickens within set parameters of thyroid hormone levels, the triiodothyronine (T3)/thyroxine (T4) ratio, and weight gain. MATERIALS AND METHODS: This study used four Pelung chickens aged 14 months. Control data consisted of data before treatment (day 0). Shell powder of A. granosa was administered for 56 days. Body weight (BW) was measured, and blood samples were drawn. In addition, the sounds from Pelung chickens were recorded once every 2 weeks on days 14, 28, 42, and 56. The blood samples were analyzed using the enzyme immunoassay method to determine testosterone, T3, and T4 levels. The sounds of Pelung chickens were recorded using the Hagemon touch method and analyzed using the Audacity application. RESULTS: The results showed a significant difference (p<0.05) in the levels of testosterone, BW, and sound frequency after the administration of A. granosa shell powder, but the administration did not have a significant effect on the levels of T4 and T3 hormones (thyroid hormone), T3/T4 ratio, or sound duration. The testosterone content and BW of Pelung chickens increased from day 14 to day 56, whereas T3 was indicative of the same profile as the T4 hormone. However, T3 and T4 levels and the T3/T4 ratio decreased and increased, respectively. The sound frequency of Pelung chickens increased from day 0 to day 14, although sound duration decreased. CONCLUSION: The administration of A. granosa shell powder at 0.9 mg/kg BW orally could improve the metabolism, testosterone level, BW, and sound frequency of Pelung chickens.

Heme Oxygenase-1 Inhibits HLA Class I Antibody-Dependent Endothelial Cell Activation.

Antibody-mediated rejection (AMR) is a key limiting factor for long-term graft survival in solid organ transplantation. Human leukocyte antigen (HLA) class I (HLA I) antibodies (Abs) play a major role in the pathogenesis of AMR via their interactions with HLA molecules on vascular endothelial cells (ECs). The antioxidant enzyme heme oxygenase (HO)-1 has anti-inflammatory functions in the endothelium. As complement-independent effects of HLA I Abs can activate ECs, it was the goal of the current study to investigate the role of HO-1 on activation of human ECs by HLA I Abs. In cell cultures of various primary human macro- and microvascular ECs treatment with monoclonal pan- and allele-specific HLA I Abs up-regulated the expression of inducible proinflammatory adhesion molecules and chemokines (vascular cell adhesion molecule-1 [VCAM-1], intercellular cell adhesion molecule-1 [ICAM-1], interleukin-8 [IL-8] and monocyte chemotactic protein 1 [MCP-1]). Pharmacological induction of HO-1 with cobalt-protoporphyrin IX reduced, whereas inhibition of HO-1 with either zinc-protoporphyrin IX or siRNA-mediated knockdown increased HLA I Ab-dependent up-regulation of VCAM-1. Treatment with two carbon monoxide (CO)-releasing molecules, which liberate the gaseous HO product CO, blocked HLA I Ab-dependent EC activation. Finally, in an in vitro adhesion assay exposure of ECs to HLA I Abs led to increased monocyte binding, which was counteracted by up-regulation of HO-1. In conclusion, HLA I Ab-dependent EC activation is modulated by endothelial HO-1 and targeted induction of this enzyme may be a novel therapeutic approach for the treatment of AMR in solid organ transplantation.

PECAM-1-dependent heme oxygenase-1 regulation via an Nrf2-mediated pathway in endothelial cells.

The antioxidant enzyme heme oxygenase (HO)-1, which catalyses the first and rate-limiting step of heme degradation, has major anti-inflammatory and immunomodulatory effects via its cell-type-specific functions in the endothelium. In the current study, we investigated whether the key endothelial adhesion and signalling receptor PECAM-1 (CD31) might be involved in the regulation of HO-1 gene expression in human endothelial cells (ECs). To this end PECAM-1 expression was down-regulated in human umbilical vein ECs (HUVECs) by an adenoviral vector-based knockdown approach. PECAM-1 knockdown markedly induced HO-1, but not the constitutive HO isoform HO-2. Nuclear translocation of the transcription factor NF-E2-related factor-2 (Nrf2), which is a master regulator of the inducible antioxidant cell response, and intracellular levels of reactive oxygen species (ROS) were increased in PECAM-1-deficient HUVECs, respectively. PECAM-1-dependent HO-1 regulation was also examined in PECAM-1 over-expressing Chinese hamster ovary and murine L-cells. Endogenous HO-1 gene expression and reporter gene activity of transiently transfected luciferase HO-1 promoter constructs with Nrf2 target sequences were decreased in PECAM-1 over-expressing cells. Moreover, a regulatory role of ROS for HO-1 regulation in these cells is demonstrated by studies with the antioxidant N-acetylcysteine and exogenous hydrogenperoxide. Finally, direct interaction of PECAM-1 with a native complex of its binding partner NB1 (CD177) and serine proteinase 3 (PR3) from human neutrophils, markedly induced HO-1 expression in HUVECs. Taken together, we demonstrate a functional link between HO-1 gene expression and PECAM-1 in human ECs, which might play a critical role in the regulation of inflammation.

Antibodies against dengue virus nonstructural protein-1 induce heme oxygenase-1 via a redox-dependent pathway in human endothelial cells.

Heme oxygenase (HO)-1, the inducible isoform of the first and rate-limiting enzyme of heme degradation, affords anti-inflammatory protection via its cell-type-specific effects in endothelial cells (ECs). In dengue hemorrhagic fever (DHF), which is the life-threatening form of dengue virus (DV) infection, endothelial interactions of cross-reactive antibodies against the DV nonstructural glycoprotein-1 (NS1) are associated with endothelial dysfunction. In this study, we investigated whether anti-NS1 antibodies might regulate HO-1 gene expression in human ECs. Serum from DHF patients with high anti-NS1 titers and a monoclonal anti-NS1 antibody upregulated HO-1 gene expression in human umbilical vein ECs, which was blocked by purified NS1 antigen. Immunoprecipitation studies showed that anti-NS1 antibodies specifically bound to the oxidoreductase protein disulfide isomerase (PDI) on ECs. Moreover, anti-NS1-mediated HO-1 induction was reduced by inhibition of PDI enzyme activity. Reactive oxygen species, which were generated by NADPH oxidase and in turn activated the phosphatidylinositol 3-kinase (PI3K)/Akt cascade, were involved in this upregulation of HO-1 gene expression. Finally, apoptosis of ECs caused by anti-NS1 antibodies was increased by pharmacological inhibition of HO-1 enzyme activity. In conclusion, HO-1 gene expression is upregulated by anti-NS1 antibodies via activation of a redox-dependent PDI/PI3K/Akt-mediated pathway in human ECs.