Functional Expression in Escherichia coli of the Disulfide-Rich Sea Anemone Peptide APETx2, a Potent Blocker of Acid-Sensing Ion Channel 3

Acid-sensing ion channels (ASICs) are proton-gated sodium channels present in the central and peripheral nervous system of chordates. ASIC3 is highly expressed in sensory neurons and plays an important role in inflammatory and ischemic pain. Thus, specific inhibitors of ASIC3 have the potential to be developed as novel analgesics. APETx2, isolated from the sea anemone Anthopleura elegantissima, is the most potent and selective inhibitor of ASIC3-containing channels. However, the mechanism of action of APETx2 and the molecular basis for its interaction with ASIC3 is not known. In order to assist in characterizing the ASIC3-APETx2 interaction, we developed an efficient and cost-effective Escherichia coli periplasmic expression system for the production of APETx2. NMR studies on uniformly (13)C/(15)N-labelled APETx2 produced in E. coli showed that the recombinant peptide adopts the native conformation. Recombinant APETx2 is equipotent with synthetic APETx2 at inhibiting ASIC3 channels expressed in Xenopus oocytes. Using this system we mutated Phe15 to Ala, which caused a profound loss of APETx2's activity on ASIC3. These findings suggest that this expression system can be used to produce mutant versions of APETx2 in order to facilitate structure-activity relationship studies.     

The correlation between the endometrial integrins and osteopontin expression with pinopodes development in ovariectomized mice in response to exogenous steroids hormones

Background: The ovariectomized animals are good models to evaluate the effect of different steroid hormone treatments on implantation events and the pattern of integrin expression. Therefore, this study was performed to compare the expression of integrins and osteopontin (OPN) in correlation with pinopode development in ovariectomized mice endometrium which was subjected to steroid hormones. Methods: Ovariectomized mice were subjected to estrogen, progesterone and estrogen-progesterone hormones. Their uterine horns were evaluated for integrin expression by immunohistochemistry and real-time RT-PCR and for pinopode development by transmission and scanning electron microscopic studies. Results: No immunostaining for integrin and OPN molecules were detected in the endometrium of non-ovariectomized mice except in metestrus phase. The α4 and β1 integrin genes were expressed in all phases of estrous cycle. In ovariectomized mice, no reaction was detected in the endometrium of control, sham and estrogen-treated groups, but in both progesterone-treated groups, all examined genes were expressed. There was not any correlation between pinopodes and integrin expression in ovariectomized mice. Conclusion: The progesterone is more effective on endometrial integrin expression than estrogen and differences in the expression pattern of integrins reflect their important and different roles in embryo implantation. The pinopodes may have minor effect in mice implantation or have some delay in their expressions in ovariectomized mice which were subjected to exogenous hormones. Key Words: Endometrium, Estrogen, Integrin, Ovariectomized mice, Osteopontin (OPN)     

Improvement of growth performance,digestive enzymes and body composition of Persian sturgeon (Acipenser persicus) following feeding on probiotics: Bacillus licheniformis,Bacillus subtilis and Saccharomyces cerevisiae

The individual and combinational effects of probiotics on Persian sturgeon (Acipenser persicus) were evaluated through assessing the growth performance, proximate body composition, digestive enzymes and intestinal morphology. On the basis of feeding with experimental diets, the fish were assigned into four groups denoted as follows: control (basal diet), D‐Pro (basal diet + Bacillus subtilis and Bacillus licheniformis), D‐Pro/Yeast (basal diet + B. subtilis and B. licheniformis + Saccharomyces cerevisiae), Yeast (basal diet + S. cerevisiae). Maximum mean weight and length were measured for the fish fed with D‐pro regime. weight gain (WG), body weight gain (BWI%),  feed efficiency (FE%), protein efficiency rate (PER), lipid efficiency rate (LER), and condition factor (CF) demonstrated the highest values for the fish fed with the diets containing D‐Pro and/or D‐Pro/yeast. LER and food conversion ratio (FCR) showed the highest performance for the fish received the D‐Pro diet. The diets containing D‐Pro exhibited higher performance in enhancing the activity of protease and lipase than the one supplemented only with the yeast, whereas the regimes treated with the yeast exhibited higher amylase activity. The fish received D‐Pro exhibited the highest protein and fat contents, yet did the lowest contents of moisture and total ash. Taken together, the findings revealed that the dietary administration of the D‐Pro and D‐Pro/Yeast could improve the growth performance of A. persicus.     

Computational Metabolomics Tools Reveal Subarmigerides,Unprecedented Linear Peptides from the Marine Sponge Holobiont Callyspongia subarmigera

A detailed examination of a unique molecular family, restricted to the Callyspongia genus, in a molecular network obtained from an in-house Haplosclerida marine sponge collection (including Haliclona, Callyspongia, Xestospongia, and Petrosia species) led to the discovery of subarmigerides, a series of rare linear peptides from Callyspongia subarmigera, a genus mainly known for polyacetylenes and lipids. The structure of the sole isolated peptide, subarmigeride A (1) was elucidated through extensive 1D and 2D NMR spectroscopy, HRMS/MS, and Marfey’s method to assign its absolute configuration. The putative structures of seven additional linear peptides were proposed by an analysis of their respective MS/MS spectra and a comparison of their fragmentation patterns with the heptapeptide 1. Surprisingly, several structurally related analogues of subarmigeride A (1) occurred in one distinct cluster from the molecular network of the cyanobacteria strains of the Guadeloupe mangroves, suggesting that the true producer of this peptide family might be the microbial sponge-associated community, i.e., the sponge-associated cyanobacteria.     

Effects of a prebiotic, Immunogen®, on feed utilization, body composition, immunity and resistance to Aeromonas hydrophila infection in the common carp Cyprinus carpio (Linnaeus) fingerlings

This study evaluated the effects of a commercial prebiotic, Immunogen, on feed utilization, growth rate, immunity promotion and carcass composition of the common carp Cyprinus carpio fingerlings. The fingerlings were adopted for 2 weeks and then reared in triplicate groups in 250-l tanks (n = 15 per tank with average initial weights of 11.12 ± 0.55 g). The fish fed on five isonitrogenous and isoenergetic experimental diets containing different levels of Immunogen (0, 0.5, 1, 1.5 and 2.5 g prebiotic/kg diet) to apparent satiation thrice a day for 8 weeks. Weight gain showed no differences among the groups fed different Immunogen levels. Both feed efficiency ratio and protein efficiency ratio significantly (p < 0.05) increased with increasing Immunogen levels from 0.5 to 1.5 g/kg diet. The highest protein content (p < 0.05) was found in the fish fed a diet containing 2.5 g/kg prebiotic. Haematological parameters and plasma total protein concentration were also significantly higher (p < 0.05) in the fingerlings fed diets containing 1.5 and 2.5 g/kg prebiotic in relation to the control. The control fish contained the highest mean of total bacterial counts. The lowest mean (p < 0.05) of total bacterial counts was observed in the fish fed the diet containing 2.5 g/kg Immunogen. The present study reveals that a dietary Immunogen supplementation from 1 to 1.5 g/kg is capable to improve the feed efficiency and growth performance of C. carpio fingerlings as well as their resistance to A. hydrophila infection.     

Bending moment resistance of corner joints constructed with spline under diagonal tension and compression

We determined the effects of the penetration depth and spline material and composite material type as well as joining method on bending moment resistance under diagonal compression and tension in common wood panel structures. Composite materials were laminated medium density fiberboard (MDF) and particle board. Joining methods were buttand miter types. Spline materials were high density fiber board (HDF).The penetration depths of plywood, wood (Carpinus betolus) and splinewere 8, 11 and 14 mm. The results showed that in both diagonal compression and tension, MDF joints are stronger than particle board joints,and the bending moment resistance under compression is higher compared with that in tension. The highest bending moment resistance under tension was shown in MDF, butt joined using plywood spline with 8 mm penetration depth, whereas under compression bending moment resistance was seen in MDF, miter joined with the HDF spline of 14 mm penetration depth.     

Arthroscopic approaches to the fetlock joint of adult cattle: A cadaver study

The objective of the present study was to describe the arthroscopic anatomy of the bovine fetlock joint using one palmar/plantar and three dorsal joint approaches. A comparative anatomic, ultrasonographic and arthroscopic study using 20 cadaveric feet from 13 non-lame adult dairy cows was performed. Arthroscopy was accomplished using a rigid arthroscope to view the synovial cavities with their synovial villi and parts of the following structures: the distal ends of the metacarpal/metatarsal III/IV bones with their trochleae and sagittal ridges, synovial grooves, the articular surfaces of the proximal sesamoid bones, the proximal aspects of the first phalanges, the lateral and medial collateral ligaments, the suspensory ligament and the interdigital ligaments as parts of the interosseus muscle, the cruciate sesamoidean ligaments, the communication site between the lateral and medial pouch in the palmar/plantar area, and dorsally the septum between the lateral and the medial pouch. The technique allowed a good overall view of most relevant structures in the sound cadaver joint. Further investigations are warranted to evaluate the diagnostic, therapeutic and prognostic applications of these techniques in the treatment of septic arthritis.     

Partial replacement of fishmeal with corn protein concentrate in diets for rainbow trout (Oncorhynchus mykiss): Effects on growth performance,physiometabolic responses,and fillet quality

An 8‐week feeding trial was run to investigate the replacement of fishmeal with corn protein concentrate (CPC; Empyreal^®) in rainbow trou (100.5 ± 2.3 g) diet. Fishmeal was increasingly replaced with 0 (Control), 30 (CPC₃), 60 (CPC₆), 90 (CPC₉) and 120 (CPC₁₂) g/kg CPC. The results showed that the substitution of fishmeal with CPC up to 90 g/kg did not cause significant differences in the growth indices as compared to the control group, while further replacement resulted in the reduction of the growth performance. The fillet crude protein showed no significant difference between the control diet and diets containing up to 90 g/kg CPC (p > .05). Serum biochemical parameters such as albumin and glucose did not change notably, while the highest levels of alanine aminotransferase and lactate dehydrogenase were seen in the CPC₁₂ group (p < .05). Also, serum lysozyme activity was increased with the elevation of dietary CPC up to 90 g/kg (p < .05). The redness and yellowness of the fillet were markedly elevated with the dietary incorporation of CPC (p < .05). Taken together, the optimum range of fishmeal substitution with CPC was 81.0–82.2 g/kg CPC in rainbow diet based on the broken‐line regression analysis.     

Colloidal stability in some tropical soils of southeastern Nigeria as affected by iron and aluminium oxides

The stability of microaggregates in soils as opposed to its dispersion is a very important soil phenomenon that checks degradation arising from unguided tillage and soil erosion. Ten soils from southeastern Nigeria were sampled from their typical A and B horizons for the study. The aim was to identify the extent of colloidal stability of the soils and the forms of Fe and Al oxides in the soils contributing to their stability. The soils are mostly Ultisols and Inceptisols formed on sandstones and shale parent materials. The soils are low in soil basic cations including the soil organic carbon (SOC). The major clay mineral is kaolinite while the soil is acid in reaction. The various forms of soil Fe and Al oxides are high with the total forms of Fe and Al being most dominant and > dithionite extracted Fe and Al > oxalate extracted Fe and Al > pyrophosphate extracted Fe and Al. The water-dispersible clay and silt (WDC) and (WDSi) which are index of dispersion in most soils are low to medium thus reflecting in the low to medium dispersion ratio (DR). The clay flocculation index (CFI) and aggregated silt + clay (ASC) were moderate to high implying the high potential stability of the soils. Soil organic carbon did not seem to be contributing much to the stability of the microaggregates while oxalate and pyrophosphate extractable Fe (Fe_ox, Fe_p) and to some extent total Al (Al_t) were among the different forms of oxides that act as aggregating agents. We propose here that rather than SOC acting as a disaggregating agent in the soils, it might have acted in association with these oxides in a linkage or bridge such as C–P–OM–C to ensure stability of the soils.