Analytical monitoring of alcoholic fermentation using NIR spectroscopy

Alcoholic fermentation under Saccharomyces cerevisiae yeasts is governed largely by glucose uptake, biomass formation, ethanol and glycerin production, and acidification. In this work, PLS calibration models were developed with a view to determining these analytical parameters from near infrared spectra and analytical data provided by the corresponding reference methods. The models were applied to a set of samples obtained from various fermentation processes. The glucose, ethanol, and biomass values predicted by the models exhibited a high correlation with those provided by the reference method.     

Dietary influence of kefir on microbial activities in the mouse bowel

AIMS: In this work the microflora present in kefir, a fermented milk product, was studied together with the effect of kefir administration on different groups of indigenous bacteria of mouse bowel. METHODS AND RESULTS: Kefir microflora was composed of lactic acid bacteria, acetic acid bacteria and yeasts. Yeast population was composed of Saccharomyces cerevisiae, S. unisporus, Candida kefir, Kluyveromyces marxianus and K. lactis. The streptococci levels in kefir treated mice increased by 10-fold and the levels of sulfite-reducing clostridia decreased by 100-fold. The number of lactic acid bacteria increased significantly. CONCLUSIONS: The administration of kefir significantly increased the lactic acid bacteria counts in the mucosa of the bowel. Ingestion of kefir specifically lowered microbial populations of Enterobacteriaceae and clostridia. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first long-term study about the effects of the kefir administration on the intestinal microflora of mice.     

A phytosociological survey of the boreal forest (shape Vaccinio-Piceetea) in North America

A survey of syntaxa of vegetation of North American boreal forests (class Vaccinio-Piceetea) is presented. This phytosociological survey, carried out combining the Braun-Blanquet method with numerical syntaxonomical analyses (cluster and correspondence analysis), describes the associations of the North American boreal forests, which have several species, varieties or vicariant species in common with their Eurasian counterparts, and can be placed in the class Vaccinio-Piceetea. By means of tabular and multivariate analyses, 2084 North American relevés were compared with 3273 relevés from European, Japanese and Korean boreal forest, to describe and typify 4 orders, 10 alliances and 37 associations. Diagnostic tables, ordination, clustering, and climatic, edaphic and biogeographical data were used to show floristic affinities among these syntaxa and interpret their distribution areas. Syntaxa were briefly characterized by their floristic composition, physiognomy, succession, zonation, and biogeographical distribution.     

Purification of Cu, Zn-Superoxide Dismutase Isoenzymes from Fish Liver: Appearance of New Isoforms as a Consequence of Pollution

Liver cell-free extracts of fish (Mugil sp.) from polluted environments show new Cu, Zn-SOD isoenzymes when analyzed by polyacrylamide gel electrophoresis or isoelectrofocusing followed by in situ staining for SOD activity. The most active isoenzymes, with pI 6.1 and 5.1, were present both in control and problem samples while the isoenzymes of intermediate pI value showed significant differences. Fish from control areas showed three intermediate isoenzymes with pI 5.7, 5.5 and 5.4 (the last one quite faint) while polluted animals showed three bands of pI 5.9, 5.45 and 5.35, this last very intense. To further characterize their utility as biomarkers, Cu, Zn-SOD isoenzymes from polluted fish livers were purified to homogeneity. Five superoxide dismutase peaks were purified, named thereafter I (pI 6.1) to V (pI 5.1) respectively. Isoenzymes I and V displayed the highest specific activity. Upon incubation with moderate H₂O₂ concentrations, pure isoenzyme I yielded more acidic bands with pI 5.5, 5.45 and 5.35, this last being predominant. The pure isoenzyme V generated only a new band of pI 5.0. Concomitant with oxidation, the activity of peaks I and V was lost in a H₂O₂ concentration-dependent manner. The pattern of the new acidic bands generated upon the oxidixing treatment of isoenzyme I closely resembles that observed in crude extracts from polluted animals.     

Mortality risk prediction of high-sensitivity C-reactive protein in suspected acute coronary syndrome: A cohort study

BackgroundThere is limited evidence on the use of high-sensitivity C-reactive protein (hsCRP) as a biomarker for selecting patients for advanced cardiovascular (CV) therapies in the modern era. The prognostic value of mildly elevated hsCRP beyond troponin in a large real-world cohort of unselected patients presenting with suspected acute coronary syndrome (ACS) is unknown. We evaluated whether a mildly elevated hsCRP (up to 15 mg/L) was associated with mortality risk, beyond troponin level, in patients with suspected ACS.Methods and findingsWe conducted a retrospective cohort study based on the National Institute for Health Research Health Informatics Collaborative data of 257,948 patients with suspected ACS who had a troponin measured at 5 cardiac centres in the United Kingdom between 2010 and 2017. Patients were divided into 4 hsCRP groups (<2, 2 to 4.9, 5 to 9.9, and 10 to 15 mg/L). The main outcome measure was mortality within 3 years of index presentation. The association between hsCRP levels and all-cause mortality was assessed using multivariable Cox regression analysis adjusted for age, sex, haemoglobin, white cell count (WCC), platelet count, creatinine, and troponin.Following the exclusion criteria, there were 102,337 patients included in the analysis (hsCRP <2 mg/L (n = 38,390), 2 to 4.9 mg/L (n = 27,397), 5 to 9.9 mg/L (n = 26,957), and 10 to 15 mg/L (n = 9,593)). On multivariable Cox regression analysis, there was a positive and graded relationship between hsCRP level and mortality at baseline, which remained at 3 years (hazard ratio (HR) (95% CI) of 1.32 (1.18 to 1.48) for those with hsCRP 2.0 to 4.9 mg/L and 1.40 (1.26 to 1.57) and 2.00 (1.75 to 2.28) for those with hsCRP 5 to 9.9 mg/L and 10 to 15 mg/L, respectively. This relationship was independent of troponin in all suspected ACS patients and was further verified in those who were confirmed to have an ACS diagnosis by clinical coding. The main limitation of our study is that we did not have data on underlying cause of death; however, the exclusion of those with abnormal WCC or hsCRP levels >15 mg/L makes it unlikely that sepsis was a major contributor.ConclusionsThese multicentre, real-world data from a large cohort of patients with suspected ACS suggest that mildly elevated hsCRP (up to 15 mg/L) may be a clinically meaningful prognostic marker beyond troponin and point to its potential utility in selecting patients for novel treatments targeting inflammation.Trial registrationClinicalTrials.gov - {"type":"clinical-trial","attrs":{"text":"NCT03507309","term_id":"NCT03507309"}}NCT03507309

Amit Kaura and colleagues investigate whether mildly elevated high sensitivity C-reactive protein is associated with mortality risk in patients with suspected acute coronary syndromes.     

Cortical amino acid neurotransmitter release is altered by CCK perfused in frontal region of unrestrained aged rats

The purpose of this study was to investigate in the aged animal the functional interaction between cholecystokinin (CCK) and amino acid neurotransmitter activity in the frontal cortex, a structure of importance in age-related disabilities. Guide cannula for repeated push-pull perfusion were implanted bilaterally in the superficial frontal cortex of male Sprague-Dawley rats. Two groups of animals were selected on the basis of their age at the time of stereotaxic surgery: 90 days and two years. Following post-operative recovery, an artificial CSF solution was perfused repeatedly within the cortex of each animal for a 5.0 min interval. The rate of perfusion was 25 microliters/min and a 5.0 min period elapsed between the collection of each sample of perfusate. After the initial control perfusions, CCK octapeptide was incorporated in a concentration of 6.0 or 18.0 ng/microliter in the CSF and perfused for 5.0 min under identical conditions. Each sample of perfusate was assayed by high performance liquid chromatography with electrochemical detection (HPLC-EC) for its content of glutamate (Glu), aspartate (Asp), glutamine (Gln), glycine (Gly), taurine (Tau) and gamma-amino-butyric acid (GABA) with homoserine used as an internal standard. Although CCK in the lower 6.0 ng/microliter concentration failed to alter significantly the profile of amino acids in the frontal cortex, the higher 18.0 ng/microliter solution of CCK enhanced the efflux of Glu as well as Asp, but only in the aged rats. Both concentrations of CCK tended also to augment the release of Gln in the older animals but these changes were not statistically significant. Both Gly and Tau were unaffected by CCK in either dose in both the young and old groups. GABA was not detectable in any of the samples of perfusate throughout the experiments. These results suggest that CCK-8 exerts a selective effect on amino acid neurotransmitter activity in the frontal cortex which is clearly age-dependent. In the older animal, this sensitivity of the cortical cells to CCK may reflect a functional attribute of the peptide in the aging process.     

Molecular and kinetic characterization and cell type location of inducible nitric oxide synthase in fish

We have found conclusive evidence for inducible nitric oxide synthase (iNOS) activity in rainbow trout (Oncorhynchus mykiss) tissue by means of biochemical, immunohistochemical, and immunoblotting analyses. This Ca(2+)-independent enzyme uses L-arginine to produce nitric oxide and L-citrulline. It was significantly inhibited by the L-arginine analogs N(omega)-monomethyl-L-arginine and N(G)-nitro-L-arginine methyl ester. Kinetic analyses showed typical Michaelian behavior with no evidence of cooperative effects. The specific activities of the liver and head kidney enzymes were 27 and 106 pmoles. min(-1). mg protein(-1), respectively, with similar values for K(m) (11 microM), all of which correspond well with the values for other previously characterized iNOS. Western blot analyses revealed a single band of M(R) = 130 kDa tested with an iNOS antiserum. At the ultrastructural level, cells with NADPH-diaphorase activity and iNOS immunoreactivity were identified as being heterophilic granulocytes in head kidney tissue and neutrophils and macrophages in hepatic tissue. The presence of an iNOS isoform in these fish tissues implies that these cells are capable of generating nitric oxide, thus pointing to the potential role of this enzyme in fish defense mechanisms.     

Phosphorylation of osteopontin is required for inhibition of vascular smooth muscle cell calcification

Osteopontin (OPN) is a non-collagenous, glycosylated phosphoprotein associated with biomineralization in osseous tissues, as well as ectopic calcification. We previously reported that osteopontin was co-localized with calcified deposits in atherosclerotic lesions, and that osteopontin potently inhibits calcium deposition in a human smooth muscle cell (HSMC) culture model of vascular calcification. In this report, the role of phosphorylation in osteopontin's mineralization inhibitory function was examined. The ability of OPN to inhibit calcification completely depended on post-translational modifications, since bacteria-derived recombinant OPN did not inhibit HSMC mineralization. Following casein kinase II treatment, phosphorylated OPN (P-OPN) dose-dependently inhibited calcification of HSMC cultured in vitro about as effectively as native OPN. The inhibitory effect of osteopontin depended on the extent of phosphorylation. To determine the specific structural domains of OPN important for inhibition of calcification, we compared OPN fragments (N-terminal, C-terminal, and full-length), and compared the inhibitory effect of both phosphorylated and non-phosphorylated fragments. While none of the non-phosphorylated OPN fragments effected calcification, P-OPN caused dose dependent inhibition of HSMC calcification. P-OPN was treated with alkaline phosphatase to create dephosphorylated OPN. Dephosphorylated OPN did not have an inhibitory effect on calcification. The expression of OPN mRNA and P-OPN secretion by HSMC were decreased in a time-dependent manner during culture calcification. These results indicate that phosphorylation is required for the inhibitory effect of OPN on HSMC calcification, and that regulation of OPN phosphorylation represents one way in which mineralization may be controlled by cells.