Angiotensin I-converting enzyme (ACE) inhibitory activities of sardinelle (Sardinella aurita) by-products protein hydrolysates obtained by treatment with microbial and visceral fish serine proteases

The angiotensin I-converting enzyme (ACE) inhibitory activities of protein hydrolysates prepared from heads and viscera of sardinelle (Sardinella aurita) by treatment with various proteases were investigated. Protein hydrolysates were obtained by treatment with Alcalase^®, chymotrypsin, crude enzyme preparations from Bacillus licheniformis NH1 and Aspergillus clavatus ES1, and crude enzyme extract from sardine (Sardina pilchardus) viscera. All hydrolysates exhibited inhibitory activity towards ACE. The alkaline protease extract from the viscera of sardine produced hydrolysate with the highest ACE inhibitory activity (63.2 ± 1.5% at 2 mg/ml). Further, the degrees of hydrolysis and the inhibitory activities of ACE increased with increasing proteolysis time. The protein hydrolysate generated with alkaline proteases from the viscera of sardine was then fractionated by size exclusion chromatography on a Sephadex G-25 into eight major fractions (P₁–P₈). Biological functions of all fractions were assayed, and P₄ was found to display a high ACE inhibitory activity. The IC₅₀ values for ACE inhibitory activities of sardinelle by-products protein hydrolysates and fraction P₄ were 1.2 ± 0.09 and 0.81 ± 0.013 mg/ml, respectively. Further, P₄ showed resistance to in vitro digestion by gastrointestinal proteases. The amino acid analysis by GC/MS showed that P₄ was rich in phenylalanine, arginine, glycine, leucine, methionine, histidine and tyrosine. The added-value of sardinelle by-products may be improved by enzymatic treatment with visceral serine proteases from sardine.     

Kinetic analysis of growth and sugar consumption by Lactobacillus fermentum IMDO 130101 reveals adaptation to the acidic sourdough ecosystem

The effect of pH on growth and carbohydrate metabolism of L. fermentum IMDO 130101 was investigated. Pronounced acid tolerance occurred together with marked responses in sugar metabolism due to acid stress. In accordance with the environment from which this strain was isolated, glucose and fructose metabolism remained active at low pH. Fructose was quantitatively converted into mannitol under all conditions tested, yielding an energetic advantage to the strain. Modelling of growth, sugar consumption, lactic and acetic acid production, and mannitol production of L. fermentum IMDO 130101 allowed the estimation of its basic biokinetic parameters when growing under simulated sourdough conditions. The obtained kinetic data underline the competitiveness of the strain in an acidic environment.     

Solid-Phase Synthesis of Substrate-Based Dipeptides and Heterocyclic Pseudo-dipeptides as Potential NO Synthase Inhibitors

More than 160 arginine analogues modified on the C-terminus via either an amide bond or a heterocyclic moiety (1,2,4-oxadiazole, 1,3,4-oxadiazole and 1,2,4-triazole) were prepared as potential inhibitors of NO synthases (NOS). A methodology involving formation of a thiocitrulline intermediate linked through its side-chain on a solid support followed by modification of its carboxylate group was developed. Finally, the side-chain thiourea group was either let unchanged, S-alkylated (Me, Et) or guanidinylated (Me, Et) to yield respectively after TFA treatment the corresponding thiocitrulline, S-Me/Et-isothiocitrulline and N-Me/Et-arginine substrate analogues. They all were tested against three recombinant NOS isoforms. Several compounds containing a S-Et- or a S-Me-Itc moiety and mainly belonging to both the dipeptide-like and 1,2,4-oxadiazole series were shown to inhibit nNOS and iNOS with IC₅₀ in the 1–50 μM range. Spectral studies confirmed that these new compounds interacted at the heme active site. The more active compounds were found to inhibit intra-cellular iNOS expressed in RAW264.7 and INS-1 cells with similar efficiency than the reference compounds L-NIL and SEIT.     

Radiation-grafted hydrogels for biomaterial applications as studied by the ESCA technique

Electron spectroscopy for chemical analysis (ESCA) was used to study the surface composition of several radiation-grafted polymers in both the dry and hydrated (frozen at 160°K) states. Poly(2-hydroxyethyl methacrylate) (HEMA) and polyacrylamide, both hydrophilic polymers, were readily detected in the hydrated or dehydrated states when grafted to polyethylene substrates. For silicone rubber substrates, both grafts were observed on the hydrated surface but were significantly decreased in surface concentration upon dehydration. For grafts on a polyester-urethane, acrylamide was not a major constituent of either the dry or hydrated surface, while HEMA appeared to increase in abundance upon drying. The amount of the hydrophobic poly(ethyl methacrylate) found on the graft surface depended upon the substrate polymer used, but the surface abundance of poly(ethyl methacrylate) was not affected by drying. These results were considered in terms of polar group orientation, polymer chain mobility, substrate permeability, and the limitations of the ESCA technique. The implications of these results with respect to the use of radiation-grafted hydrophilic polymers for biomedical applications are also discussed.     

Resistance of stainless steel CHS columns based on cross-section deformation capacity

A conceptually new structural design approach has recently been proposed by the authors to predict the resistance of stainless steel members subjected to various types of loading with cross-sections formed from thin flat plates including angles, channels, lipped channels, I-sections and rectangular hollow sections (RHS). The proposed method does not follow the traditional cross-section classification approach, which primarily relies on the assumption of a bilinear, elastic-perfectly-plastic material model. Instead, deformation capacity of a cross-section is determined directly from the local buckling characteristics of the constituent plate elements. This is then used to obtain the corresponding local buckling stress utilising an appropriate material model. This basic concept is extended herein to predict compression resistance of stainless steel columns with circular hollow sections (CHS). Available test and finite element (FE) results have been used to develop the basic design equation to predict the compression resistance of cross-sections and to propose column curves to determine flexural buckling resistances. The predicted resistances have been compared to those obtained using the current Eurocode; the predictions are significantly more accurate and more consistent than those given by the existing Eurocode.     

Reversible electron-transfer reactions within a nanoscale metal oxide cage mediated by metallic substrates

Transition metal oxides exhibit a rich collection of electronic properties and have many practical applications in areas such as catalysis and ultra-high-density magnetic data storage. Therefore the development of switchable molecular transition metal oxides has potential for the engineering of single-molecule devices and nanoscale electronics. At present, the electronic properties of transition metal oxides can only be tailored through the irreversible introduction of dopant ions, modifying the electronic structure by either injecting electrons or core holes. Here we show that a molybdenum(VI) oxide 'polyoxometalate' molecular nanocluster containing two embedded redox agents is activated by a metallic surface and can reversibly interconvert between two electronic states. Upon thermal activation two electrons are ejected from the active sulphite anions and delocalized over the metal oxide cluster cage, switching it from a fully oxidized state to a two-electron reduced state along with the concomitant formation of an S-S bonding interaction between the two sulphur centres inside the cluster shell.     

Mass Spectrometry-Based Proteomics Reveal Alcohol Dehydrogenase 1B as a Blood Biomarker Candidate to Monitor Acetaminophen-Induced Liver Injury

Acute liver injury (ALI) is a severe disorder resulting from excessive hepatocyte cell death, and frequently caused by acetaminophen intoxication. Clinical management of ALI progression is hampered by the dearth of blood biomarkers available. In this study, a bioinformatics workflow was developed to screen omics databases and identify potential biomarkers for hepatocyte cell death. Then, discovery proteomics was harnessed to select from among these candidates those that were specifically detected in the blood of acetaminophen-induced ALI patients. Among these candidates, the isoenzyme alcohol dehydrogenase 1B (ADH1B) was massively leaked into the blood. To evaluate ADH1B, we developed a targeted proteomics assay and quantified ADH1B in serum samples collected at different times from 17 patients admitted for acetaminophen-induced ALI. Serum ADH1B concentrations increased markedly during the acute phase of the disease, and dropped to undetectable levels during recovery. In contrast to alanine aminotransferase activity, the rapid drop in circulating ADH1B concentrations was followed by an improvement in the international normalized ratio (INR) within 10–48 h, and was associated with favorable outcomes. In conclusion, the combination of omics data exploration and proteomics revealed ADH1B as a new blood biomarker candidate that could be useful for the monitoring of acetaminophen-induced ALI.     

Influence of Risk Factors for Male Infertility on Sperm Protein Composition

Male infertility is a common health problem that can be influenced by a host of lifestyle risk factors such as environment, nutrition, smoking, stress, and endocrine disruptors. These effects have been largely demonstrated on sperm parameters (e.g., motility, numeration, vitality, DNA integrity). In addition, several studies showed the deregulation of sperm proteins in relation to some of these factors. This review inventories the literature related to the identification of sperm proteins showing abundance variations in response to the four risk factors for male infertility that are the most investigated in this context: obesity, diabetes, tobacco smoking, and exposure to bisphenol-A (BPA). First, we provide an overview of the techniques used to identify deregulated proteins. Then, we summarise the main results obtained in the different studies and provide a compiled list of deregulated proteins in relation to each risk factor. Gene ontology analysis of these deregulated proteins shows that oxidative stress and immune and inflammatory responses are common mechanisms involved in sperm alterations encountered in relation to the risk factors.     

Staphylococcal community of a small unit manufacturing traditional dry fermented sausages

The level and the diversity of the staphylococcal community occurring in the environment and meat products of a small unit manufacturing traditional dry fermented sausages were investigated at two seasons: winter and spring. Gram-positive cocci were enumerated and a collection of 412 Staphylococcus isolates was made. Multiplex PCR, pulse-field gel electrophoresis (PFGE) and sequencing of the sodA gene were used to identify and characterize the isolates. High counts of Staphylococcus were found in final traditional sausages, reaching about 6 log CFU/g in winter and about 8 log CFU/g in spring. In the environment, the counts varied from 2 log to 7 log/100 cm(2), the higher colonisation being observed on the surface of the drying and cold rooms, cutting tables and the butcher's block. The combination of the three methods allowed the identification of seven species of Staphylococcus in spring and five in winter. S. equorum and S. succinus dominated both in environment and in meat products, 49% and 33% of the isolates, respectively. The other identified species were in decreasing order S. saprophyticus (6%), S. xylosus (5%), S. carnosus (5%), S. simulans (1%) and S. warneri (1%). The two species S. xylosus and S. carnosus were sporadically isolated during the spring. PFGE allowed the assignment of S. equorum to eight pulsotypes showing a wide diversity among this species. But the entire environment and the meat products were dominated by one pulsotype. For S. succinus, three pulsotypes were found with one dominant mainly isolated during the spring sampling. This study highlighted the diversity of staphylococci isolated in the environment and the meat products of a small processing unit manufacturing traditional dry fermented sausages. The S. equorum and S. succinus species rarely described in meat products and never in the environment had great capacity to colonise the entire small processing unit and the meat products.