Non-exhaustive test for aerobic capacity determination in running rats

A simple and applicable method for non-exhaustive aerobic evaluation in running rats is described. Wistar rats were submitted to running test at different velocities (10, 15, 20, 25 m/min) with 48 h recovery among them. At each velocity, the rats ran two bouts of 5 min with 2 min of rest between bouts. Blood samples were collected at the end of each bout for lactate determination. For each intensity, delta lactate was calculated and using deltas obtained by four tests, an individual linear interpolation was plotted. The y-intercept of linear interpolation was the null delta lactate equivalent to the critical velocity (CV). To verify the lactate stabilization at CV, the animals were submitted to 25 min of continuous exercise (15, 20, 25 m/min), with blood collection every 5 min. The estimated CV was 16.6±0.7 m/min, with significant linear regressions (R=0.90±0.03). The rats presented maximal lactate steady state (MLSS) at 3.9±0.4 mmol/L, at 20 m/min. The CV was less than MLSS but significantly correlated with this parameter (r=0.78). This non-exhaustive test seems to be valid for the aerobic evaluation of sedentary rats and this protocol underestimates the MLSS in 20%. This test seems to be the interesting method for the evaluation of rats submitted to acute exercise or physical training.

Organic milk improves Bifidobacterium lactis counts and bioactive fatty acids contents in fermented milk

In functional dairy products, polyunsaturated fatty acids such as, conjugated linoleic acid (CLA) and alpha-linolenic acid (ALA) have been highlighted for their benefits related to prevention of some chronic diseases. In order to study the effect of type of milk (conventional vs. organic, characterized by a specific fatty acid composition), Bifidobacterium animalis subsp. lactis (BB12, B94, BL04 and HN019) counts, acidification activity and chemical composition (pH, lactose, lactic acid contents and fatty acids profile) were investigated before fermentation and after 24 h of products stored at 4 degrees C. Organic and conventional milk influenced acidification performance and bacteria counts, which was strain-dependent. Higher counts of BB12 were observed in organic milk, whereas superior counts of BL04 were found in conventional milk. Organic fermented milk showed lower levels in saturated fatty acids (FA) and higher in monounsaturated FA contents. Similarly, among bioactive FA, organic fermented milks have higher amounts of trans vaccenic acid (TVA-C18:1t), conjugated linoleic acid (CLA) and slightly higher contents of alpha-linoleic acid (ALA). (C) 2012 Elsevier Ltd. All rights reserved.

Biomedical and industrial applications of atmospheric pressure non-equilibrium plasmas

This dissertation will be focused on the characterization of an atmospheric pressure plasma jet source with an application oriented diagnostic approach and the description of processes supported by this plasma source. The plasma source investigated is a single electrode plasma jet. Schlieren images, optical emission spectra, temperature and heat flux profiles are analyzed to deeply investigate the fluid dynamic, the chemical composition and the thermal output of the plasma generated with a nanosecond-pulsed high voltage generator. The maximum temperature measured is about 45 °C and values close to the room temperature are reached 10 mm down the source outlet, ensuring the possibility to use the plasma jet for the treatment of thermosensitive materials, such as, for example, biological substrate or polymers. Electrospinning of polymeric solution allows the production of nanofibrous non-woven mats and the plasma pre-treatment of the solutions leads to the realization of defect free nanofibers. The use of the plasma jet allows the electrospinnability of a non-spinnable poly(L-lactic acid) (PLLA) solution, suitable for the production of biological scaffold for the wound dressing.

Surface modification of natural fibers using bacteria:depositing bacterial cellulose onto natural fibers to create hierarchical fiber reinforced nanocomposites

Triggered biodegradable composites made entirely from renewable resources are urgently sought after to improve material recyclability or be able to divert materials from waste streams. Many biobased polymers and natural fibers usually display poor interfacial adhesion when combined in a composite material. Here we propose a way to modify the surfaces of natural fibers by utilizing bacteria (Acetobacter xylinum) to deposit nanosized bacterial cellulose around natural fibers, which enhances their adhesion to renewable polymers. This paper describes the process of modifying large quantities of natural fibers with bacterial cellulose through their use as substrates for bacteria during fermentation. The modified fibers were characterized by scanning electron microscopy, single fiber tensile tests, X-ray photoelectron spectroscopy, and inverse gas chromatography to determine their surface and mechanical properties. The practical adhesion between the modified fibers and the renewable polymers cellulose acetate butyrate and poly(L-lactic acid) was quantified using the single fiber pullout test. © 2008 American Chemical Society.

Ruthenibacterium lactatiformans gen. nov., sp.nov., an anaerobic, lactate-producing member of the family Ruminococcaceae isolated from human faeces

Two novel strains of Gram-stain-negative, rod-shaped, obligately anaerobic, non-spore-forming, non-motile bacteria were isolated from the faeces of healthy human subjects. The strains, designated as 585-1T and 668, were characterized by mesophilic fermentative metabolism, production of d-lactic acid, succinic acid and acetic acid as end products of d-glucose fermentation, prevalence of C18 : 1 ω9, C18 : 1 ω9 aldehyde, C16 : 0 and C16 : 1 ω7c fatty acids, presence of glycine, glutamic acid, lysine, alanine and aspartic acid in the petidoglycan peptide moiety and lack of respiratory quinones. Whole genome sequencing revealed the DNA G+C content was 56.4–56.6 mol%. The complete 16S rRNA gene sequences of the two strains shared 91.7/91.6 % similarity with Anaerofilum pentosovorans FaeT, 91.3/91.2 % with Gemmiger formicilis ATCC 27749T and 88.9/88.8 % with Faecalibacterium prausnitzii ATCC 27768T. On the basis of chemotaxonomic and genomic properties it was concluded that the strains represent a novel species in a new genus within the family Ruminococcaceae , for which the name Ruthenibacterium lactatiformans gen. nov., sp. nov. is proposed. The type strain of Ruthenibacterium lactatiformans is 585-1T (=DSM 100348T=VKM B-2901T).

Características de fermentação da silagem de seis variedades de milho indicadas para a região semiárida Brasileira

Avaliaram-se as características fermentativas e a qualidade das silagens de seis variedades de milho, de ciclos precoce e superprecoce - BRS Caatingueiro, BRS Assum Preto, BR 5033 Asa Branca, BR 5028 São Francisco, Gurutuba e BRS 4103 - indicadas para a região semiárida brasileira. Foram utilizados silos experimentais, em delineamento inteiramente ao acaso, com seis tratamentos (variedades) e quatro repetições. Avaliaram-se: matéria seca (MS), matéria orgânica (MO), proteína bruta (PB), fibra em detergente neutro (FDN), fibra em detergente ácido (FDA), extrato etéreo (EE), carboidratos totais (CHO), carboidratos não fibrosos (CNF), pH, nitrogênio amoniacal como parte do nitrogênio total (N-NH3/NT), ácidos orgânicos e digestibilidade in vitro da matéria seca (DIVMS) das silagens. Os valores médios encontrados para a silagem foram: MS= 28,7%; MO= 94,9%; PB= 8,3%; FDN= 49,9%; FDA= 27,5%; EE= 3,8%; CHO= 82,7%; CNF= 32,8%; pH= 3,8; N-NH3/NT= 2,9%/NT; ácido láctico = 7,6%; ácido acético = 0,6%; ácido butírico = 0,3% e DIVMS= 57,9%. As variedades BR 5028 - São Francisco e Gurutuba destacaram-se das demais em relação ao teor de matéria seca. A variedade BRS Caatingueiro apresentou maior teor de carboidratos não fibrosos em relação às demais. As silagens de todas as variedades foram classificadas como de excelente qualidade, por apresentarem potencial para ensilagem no semiárido brasileiro

The role of γ -aminobutyric acid (Gaba) in somatic embryogenesis of Acca sellowiana Berg. (Myrtaceae)

The γ-aminobutyric acid (Gaba) is a non-protein amino acid found in prokaryotes and eukaryotes. Its role in plant development has not been fully established. This study reports a quantification of the levels of endogenous Gaba, as well as investigation of its role in different stages of somatic embryogenesis in Acca sellowiana Berg. (Myrtaceae). Zygotic embryos were used as explants and they were inoculated into the culture medium contained different concentrations of Gaba (0,2, 4, 6, 8 and 10 µM). The highest concentrations of endogenous Gaba were detected between the third and nine days after inoculation, reaching the value of 12.77 µmol.g-1FW. High frequency of somatic embryogenesis was observed in response to 10 µM Gaba. This treatment also resulted in a large number of normal embryos, and the lowest percentage of formation of fused somatic embryos, phenotypic characteristic of most deformed embryos in all treatments. Also, all treatments promoted the formation of the somatic embryos with positive characteristics of development resumption, which however did not originate the seedlings.

Effects of trans-10, cis-12 conjugated linoleic acid on gene expression and lipid metabolism of adipose tissue of growing pigs

The objective of the present study was to determine the effects of trans-10, cis-12 conjugated linoleic acid (CLA) in adipose tissue explant cultures of growing pigs on the following responses: lipogenesis (measured as rate of C-14-labeled glucose incorporation over a subsequent 2-h incubation in the presence or absence of insulin), lipolysis (release of non-esterified fatty acid over a 2-h incubation in the presence or absence of isoproterenol), activities of lipogenic enzymes, and mRNA abundance of fatty acid synthase (FAS). Adipose tissue explants from nine growing pigs (78 +/- 3 kg) were cultured in 199 medium with insulin, dexamethasone and antibiotics for 4, 12, 24, and 48 h. The treatments were 1) control: 100 mu M polyvinyl alcohol (PVA); 2) pGH: 100 ng/mL porcine growth hormone (pGH) plus 100 mu M PVA; 3) CLA200: 200 mu M trans-10, cis-12 CLA; 4) CLA50: 50 mu M trans-10, cis-12 CLA, and 5) LA: 200 mu M linoleic acid. Fatty acids were added along with PVA (2: 1), respectively, for 24 h. Explants were collected after each culture period and assayed for lipogenesis. Transcripts of FAS mRNA were quantified by real-time RT-PCR after 24 and 48 h. Lipolysis and activities of FAS, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and NADP-malate dehydrogenase were determined after 48 h. As expected, glucose incorporation was decreased (P < 0.05) in response to pGH treatment (positive control). LA had no effect on any parameter evaluated. Treatment with trans-10, cis-12 CLA decreased FAS activity (P < 0.05), but NADPH-generating enzymes were unaffected by treatments. Consistent with reduction in FAS activity, both lipid synthesis and FAS mRNA abundance were reduced with chronic CLA treatment, pGH increased baseline and stimulated lipolysis (P < 0.05) after 48 h of culture, while CLA treatment had no effect on non-esterified fatty acid release. Results of this study showed that trans-10, cis-12 CLA alters lipogenesis but has no effect on lipolysis in cultures of pig adipose tissue.

Behavior and viability of spontaneous oxidative stress-resistant Lactococcus lactis mutants in experimental fermented milk processing

Previously, we isolated two strains of spontaneous oxidative (SpOx2 and SpOx3) stress mutants of Lactococcus lactis subsp cremoris. Herein, we compared these mutants to a parental wild- type strain (J60011) and a commercial starter in experimental fermented milk production. Total solid contents of milk and fermentation temperature both affected the acidification profile of the spontaneous oxidative stress- resistant L. lactis mutants during fermented milk production. Fermentation times to pH 4.7 ranged from 6.40 h (J60011) to 9.36 h (SpOx2); V(max) values were inversely proportional to fermentation time. Bacterial counts increased to above 8.50 log(10) cfu/mL. The counts of viable SpOx3 mutants were higher than those of the parental wild strain in all treatments. All fermented milk products showed post-fermentation acidification after 24 h of storage at 4 degrees C; they remained stable after one week of storage.

Effect of different prebiotics on the fermentation kinetics, probiotic survival and fatty acids profiles in nonfat symbiotic fermented milk

The simultaneous effects of different binary co-cultures of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus rhamnosus and Bifidobacterium lactis with Streptococcus thermophilus and of different prebiotics on the production of fermented milk were investigated in this paper. In particular, we determined and compared the kinetics of acidification of milk either as such or supplemented with 4% (w/w) maltodextrin, oligofructose and polydextrose, as well as the probiotic survival, chemical composition (pH, lactose, lactic acid and protein contents), fatty acids profile and conjugate linoleic acid (CIA) content of fermented milk after storage at 4 degrees C for 24 h. Fermented milk quality was strongly influenced both by the co-culture composition and the selected prebiotic. Depending on the co-culture, prebiotic addition to milk influenced to different extent kinetic acidification parameters. All probiotic counts were stimulated by oligofructose and polydextrose, and among these B. lactis always exhibited the highest counts in all supplemented milk samples. Polydextrose addition led to the highest post-acidification. Although the contents of the main fatty acids were only barely influenced. the highest amounts of conjugated linoleic acid (38% higher than in the control) were found in milk fermented by S. thermophilus-L. acidophilus co-culture and supplemented with maltodextrin. (C) 2008 Elsevier B.V. All rights reserved.

Description of Gluconacetobacter sacchari sp. nov., a new species of acetic acid bacterium isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug

A new species of the genus Gluconacetobacter, for which the name Gluconacetobacter sacchari sp. nov. is proposed, was isolated from the leaf sheath of sugar cane and from the pink sugar-cane mealy bug, Saccharicoccus sacchari, found on sugar cane growing in Queensland and northern New South Wales, Australia, The nearest phylogenetic relatives in the alpha-subclass of the Proteobacteria are Gluconacetobacter liquefaciens and Gluconacetobacter diazotrophicus, which have 98.8-99.3% and 97.9-98.5% 16S rDNA sequence similarity, respectively, to members of Gluconacetobacter sacchari. On the basis of the phylogenetic positioning of the strains, DNA reassociation studies, phenotypic tests and the presence of the Q10 ubiquinone, this new species was assigned to the genus Gluconacetobacter. No single phenotypic characteristic is unique to the species, but the species can be differentiated phenotypically from closely related members of the acetic acid bacteria by growth in the presence of 0.01% malachite green, growth on 30% glucose, an inability to fix nitrogen and an inability to grow with the L-amino acids asparagine, glycine, glutamine, threonine and tryptophan when D-mannitol was supplied as the sole carbon and energy source. The type strain of this species is strain SRI 1794(T) (= DSM 12717(T)).

Effects of adding polyclonal antibody preparations on ruminal fermentation patterns and digestibility of cows fed different energy sources

Nine ruminally cannulated cows fed different energy sources were used to evaluate an avian-derived polyclonal antibody preparation (PAP-MV) against the specific ruminal bacteria Streptococcus bovis, Fusobacterium necrophorum, Clostridium aminophilum, Peptostreptococcus anaerobius, and Clostridium stick-landii and monensin (MON) on ruminal fermentation patterns and in vivo digestibility. The experimental design was three 3 x 3 Latin squares distinguished by the main energy source in the diet [dry-ground corn grain (CG), high-moisture corn silage (HMCS), or citrus pulp (CiPu)]. Inside each Latin square, animals received one of the feed additives per period [none (CON), MON, or PAP-MV]. Dry matter intake and ruminal fermentation variables such as pH, total short-chain fatty acids (tSCFA), which included acetate, propionate, and butyrate, as well as lactic acid and NH(3)-N concentration were analyzed in this trial. Total tract DM apparent digestibility and its fractions were estimated using chromic oxide as an external marker. Each experimental period lasted 21 d. Ruminal fluid sampling was carried out on the last day of the period at 0, 2, 4, 6, 8, 10, and 12 h after the morning meal. Ruminal pH was higher (P = 0.006) 4 h postfeeding in MON and PAP-MV groups when compared with CON. Acetate: propionate ratio was greater in PAP-MV compared with MON across sampling times. Polyclonal antibodies did not alter (P > 0.05) tSCFA, molar proportion of acetate and butyrate, or lactic acid and NH(3)-N concentration. Ruminal pH was higher (P = 0.01), 4 h postfeeding in CiPu diets compared with CG and HMCS. There was no interaction between feed additive and energy source (P > 0.05) for any of the digestibility coefficients analyzed. Starch digestibility was less (P = 0.008) in PAP-MV when compared with CON and MON. In relation to energy sources, NDF digestibility was greater (P = 0.007) in CG and CiPu vs. the HMCS diet. The digestibility of ADF was greater (P = 0.002) in CiPu diets followed by CG and HMCS. Feeding PAP-MV or monensin altered ruminal fermentation patterns and digestive function in cows; however, those changes were independent of the main energy source of the diet.

Comparison of Cross-Sectional Hardness and Transverse Microradiography of Artificial Carious Enamel Lesions Induced by Different Demineralising Solutions and Gels

The aims of this study were: (1) to correlate surface (SH) and cross-sectional hardness (CSH) with microradiographic parameters of artificial enamel lesions; (2) to compare lesions prepared by different protocols. Fifty bovine enamel specimens were allocated by stratified randomisation according to their initial SH values to five groups and lesions produced by different methods: MC gel (methylcellulose gel/lactic acid, pH 4.6, 14 days); PA gel (polyacrylic acid/lactic acid/hydroxyapatite, pH 4.8, 16 h); MHDP (undersaturated lactate buffer/methyl diphosphonate, pH 5.0, 6 days); buffer (undersaturated acetate buffer/fluoride, pH 5.0, 16 h), and pH cycling (7 days). SH of the lesions (SH(1)) was measured. The specimens were longitudinally sectioned and transverse microradiography (TMR) and CSH measured at 10- to 220-mu m depth from the surface. Overall, there was a medium correlation but non-linear and variable relationship between mineral content and root CSH. root SH(1) was weakly to moderately correlated with surface layer properties, weakly correlated with lesion depth but uncorrelated with integrated mineral loss. MHDP lesions showed the highest subsurface mineral loss, followed by pH cycling, buffer, PA gel and MC gel lesions. The conclusions were: (1) CSH, as an alternative to TMR, does not estimate mineral content very accurately, but gives information about mechanical properties of lesions; (2) SH should not be used to analyse lesions; (3) artificial caries lesions produced by the protocols differ, especially considering the method of analysis. Copyright (C) 2009 S. Karger AG, Basel

Bioactive albumin functionalized polylactic acid membranes for improved biocompatibility

Biocompatibility is a major challenge for successful application of many biomaterials. In this study the ability to coat chemically and enzymatically activated poly(L-lactic acid) (PLA) membranes with heat denatured human serum albumin to improve biocompatibility was investigated. PLA membranes hydrolyzed with NaOH or cutinase and then treated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, hydrochloride (EDAC) as a heterobifunctional cross-linker promoted the coupling single bondCOOH groups on PLA membranes and single bondNH2 groups of heat denatured human serum albumin. This resulted in increased hydrophilicity (lowest water contact angles of 43° and 35°) and highest antioxidant activity (quenching of 79 μM and 115 μM tetramethylazobisquinone (TMAMQ) for NaOH and cutinase pretreated membranes, respectively). FTIR analysis of modified PLA membranes showed new peaks attributed to human serum albumin (amide bond, NH2 and side chain stretching) appearing within 3600–3000 cm−1 and 1700–1500 cm−1 (Fig. 3). MTT studies also showed that osteoblasts-like and MC-3T3-E1 cells viability increased 2.4 times as compared to untreated PLA membranes. The study therefore shows that this strategy of modifying the surfaces of PLA polymers could significantly improve biocompatibility.

Lactacidemia: métodos de dosagem do ácido lático

A critical study of three methods for the determination of lactic acid (EDWARDS, MENDEL & GOLDSCHEIDER, MILLER & MUNTZ) is presented and some modifications are proposed. It was shown t hat more accurate results could be obtained with Edward's technic when an Iena glass filter is connected with the absorption tube. Before the dropping of the permanganate solution it is necessary to pass a current of air through the reaction flask to avoid the oxidation of the non-lactic acid substances which interfere with the reaction. The absorption tube must be maintained at 18°C during the destillation and the titration of the bisulphite binding aldehyde at 4°C. When the sample contains more than 5 mg it is useful to work with greater quantities of the bisulphite. More permanganate is consumed when the lactic acid concentration is higher. The sensivity of the method permits the titration of 0.04 mg to 5 mg of lactic acid in the sample. The calculated error of the method gave 0.018 % and the normal values for blood determined in 20 human cases averaged 10.30 mg per 100 ml (Table VI). MENDEL and GOLDSCHEIDER'S method was modified in the following details: Somogyis deproteinization was performed instead metaphosphoric acid as in the original method; to avoid the evaporation of the acetic aldehyde during the heating time with sulfuric acid a special glass stopped tube is proposed (Fig. 2). The reaction with sulfuric acid and veratrol is performed in an ice bath. Blood proteins precipitants were tried and Somogyi's lattest tecnic showed better results (Table V). Colorimetric readings were done in the PULFRICH photometer using filter S 53 and a 10 mm cup. The method is accurate within an error of 0.23 % and samples of 5 to 70 microg. could be easily determined. Normal values for human blood averaged 10.78 mg per 100 ml. More accurate results were obtained with the technic of MILLER & MUNTZ. Slight modifications were introduced: deproteinization with copper sulfate and sodium tungstate; satured p-hydroxydiphenyl solution according to KOENEMANN which is stable for 5 months when stored in the ice-box. Using the PULFRICH step-photometer the error is 0.17% with samples varying from 0.1 to 10 microg. of lactic acid. The filter employed was S 57 with the 5 mm cup. The method was adapted to 0.1 ml of blood. Normal values for human blood gave an average of 10.58 mg per 100 ml.