Expression of a hantavirus N protein and its efficacy as antigen in immune assays

Hantavirus cardiopulmonary syndrome (HCPS) has been recognized as an important public heath problem. Five hantaviruses associated with HCPS are currently known in Brazil: Juquitiba, Araraquara, Laguna Negra-like, Castelo dos Sonhos, and Anajatuba viruses. The laboratory diagnosis of HCPS is routinely carried out by the detection of anti-hantavirus IgM and/or IgG antibodies. The present study describes the expression of the N protein of a hantavirus detected in the blood sample of an HCPS patient. The entire S segment of the virus was amplified and found to be 1858 nucleotides long, with an open reading frame of 1287 nucleotides that encodes a protein of 429 amino acids. The nucleotide sequence described here showed a high identity with the N protein gene of Araraquara virus. The entire N protein was expressed using the vector pET200D and the Escherichia coli BL21 strain. The expression of the recombinant protein was confirmed by the detection of a 52-kDa protein by Western blot using a pool of human sera obtained from HCPS patients, and by specific IgG detection in five serum samples of HCPS patients tested by ELISA. These results suggest that the recombinant N protein could be used as an antigen for the serological screening of hantavirus infection.

Changes in the hormone and lipid profile of obese adolescent Saudi females with acne vulgaris

Acne vulgaris is a multifactorial disease affecting a majority of the adolescent population. The objective of this study was to test for a correlation between fasting serum lipid profiles and levels of testosterone, insulin, leptin, and interleukin 1-β (IL-1β) and the incidence of severe acne vulgaris in obese adolescent females. Four groups of adolescent females were studied: obese with acne, obese without acne, non-obese with acne, and non-obese without acne. Obese females with acne, compared to obese females without acne and non-obese subjects, had significantly higher serum triglycerides, low-density lipoprotein cholesterol and apolipoprotein-B (apo-B) (mean ± SD: 197 ± 13.7 vs 171 ± 11.5, 128 ± 8.3 vs 116 ± 7.7, 96 ± 13.7 vs 85 ± 10.3 mg/dL, respectively) but significantly lower high-density lipoprotein cholesterol and apo-A1 levels (40 ± 3.3 vs 33 ± 3.5 and 126 ± 12 vs 147 ± 13 mg/dL). Serum testosterone, insulin and leptin levels were significantly higher in obese subjects with or without acne compared to non-obese females with or without acne (3 ± 0.5 vs 2.1 ± 0.47, 15.5 ± 3.3 vs 11.6 ± 3, 0.9 ± 0.2 vs 0.6 ± 0.15 nmol/mL, respectively). Serum IL-1b was significantly elevated in obese and non-obese subjects with acne compared to subjects without acne; in those without acne, these levels were higher in obese than non-obese subjects (2.4 ± 0.2, 1.4 ± 0.1 vs 1.8 ± 0.12 and 1.3 ± 0.11 pg/mL, respectively). Our results indicate that there is a relationship between obesity (BMI >27) and acne. By early recognition, the etiology and treatment protocol of acne may prevent unwanted conditions.

Structural properties of lipid reconstructs and lipid composition of normotensive and hypertensive rat vascular smooth muscle cell membranes

Multiple cell membrane alterations have been reported to be the cause of various forms of hypertension. The present study focuses on the lipid portion of the membranes, characterizing the microviscosity of membranes reconstituted with lipids extracted from the aorta and mesenteric arteries of spontaneously hypertensive (SHR) and normotensive control rat strains (WKY and NWR). Membrane-incorporated phospholipid spin labels were used to monitor the bilayer structure at different depths. The packing of lipids extracted from both aorta and mesenteric arteries of normotensive and hypertensive rats was similar. Lipid extract analysis showed similar phospholipid composition for all membranes. However, cholesterol content was lower in SHR arteries than in normotensive animal arteries. These findings contrast with the fact that the SHR aorta is hyporeactive while the SHR mesenteric artery is hyperreactive to vasopressor agents when compared to the vessels of normotensive animal strains. Hence, factors other than microviscosity of bulk lipids contribute to the vascular smooth muscle reactivity and hypertension of SHR. The excess cholesterol in the arteries of normotensive animal strains apparently is not dissolved in bulk lipids and is not directly related to vascular reactivity since it is present in both the aorta and mesenteric arteries. The lower cholesterol concentrations in SHR arteries may in fact result from metabolic differences due to the hypertensive state or to genes that co-segregate with those that determine hypertension during the process of strain selection.

Highly sensitive C-reactive protein and male gender are independently related to the severity of coronary disease in patients with metabolic syndrome and an acute coronary event

Patients with metabolic syndrome are at high-risk for development of atherosclerosis and cardiovascular events. The objective of this study was to examine the major determinants of coronary disease severity, including those coronary risk factors associated with metabolic syndrome, during the early period after an acute coronary episode. We tested the hypothesis that inflammatory markers, especially highly sensitive C-reactive protein (hsCRP), are related to coronary atherosclerosis, in addition to traditional coronary risk factors. Subjects of both genders aged 30 to 75 years (N = 116) were prospectively included if they had suffered a recent acute coronary syndrome (acute myocardial infarction or unstable angina pectoris requiring hospitalization) and if they had metabolic syndrome diagnosed according to the National Cholesterol Education Program/Adult Treatment Panel III. Patients were submitted to a coronary angiography and the burden of atherosclerosis was estimated by the Gensini score. The severity of coronary disease was correlated (Spearman’s or Pearson’s coefficient) with gender (r = 0.291, P = 0.008), age (r = 0.218, P = 0.048), hsCRP (r = 0.256, P = 0.020), ApoB/ApoA ratio (r = 0.233, P = 0.041), and carotid intima-media thickness (r = 0.236, P = 0.041). After multiple linear regression, only male gender (P = 0.046) and hsCRP (P = 0.012) remained independently associated with the Gensini score. In this high-risk population, male gender and high levels of hsCRP, two variables that can be easily obtained, were associated with more extensive coronary disease, identifying patients with the highest potential of developing new coronary events.

Interaction between human cytomegalovirus UL136 protein and ATP1B1 protein

Interplay between the host and human cytomegalovirus (HCMV) has a pivotal role in the outcome of infection. A region (referred to as UL/b’) present in the Toledo strain of HCMV and low passage clinical isolates contains 19 additional genes, which are absent in the highly passaged laboratory strain AD169. Products of the UL/b’ genes may determine the manifestations of HCMV infection in vivo. However, little is known about the host factors, which interact with UL/b’ proteins. This study was conducted to investigate the function of the HCMV UL136 protein. By yeast two-hybrid screening, the β1 subunit of the host Na+/K+-ATPase (ATP1B1) was identified to be a candidate protein, which interacts with the HCMV UL136 protein. The interaction was further evaluated both in vitro by pull-down assay and in vivo by immunofluorescent co-localization. The results showed that the UL136 protein can interact with ATP1B1 in vitro. Co-localization of UL136-EGFP and ATP1B1-DsRed in cell membranes suggests that ATP1B1 was a partner of the UL136 protein. It can be proposed that the HCMV UL136 protein may have important roles in processes such as cell-to-cell spread, and in maintaining cell osmotic pressure and intracellular ion homeostasis during HCMV infection.

The RecX protein interacts with the RecA protein and modulates its activity in Herbaspirillum seropedicae

DNA repair is crucial to the survival of all organisms. The bacterial RecA protein is a central component in the SOS response and in recombinational and SOS DNA repairs. The RecX protein has been characterized as a negative modulator of RecA activity in many bacteria. The recA and recX genes of Herbaspirillum seropedicae constitute a single operon, and evidence suggests that RecX participates in SOS repair. In the present study, we show that the H. seropedicae RecX protein (RecX Hs) can interact with the H. seropedicaeRecA protein (RecA Hs) and that RecA Hs possesses ATP binding, ATP hydrolyzing and DNA strand exchange activities. RecX Hs inhibited 90% of the RecA Hs DNA strand exchange activity even when present in a 50-fold lower molar concentration than RecA Hs. RecA Hs ATP binding was not affected by the addition of RecX, but the ATPase activity was reduced. When RecX Hs was present before the formation of RecA filaments (RecA-ssDNA), inhibition of ATPase activity was substantially reduced and excess ssDNA also partially suppressed this inhibition. The results suggest that the RecX Hs protein negatively modulates the RecA Hs activities by protein-protein interactions and also by DNA-protein interactions.

Physicochemical and sensory profile of rice bran roasted in microwave

The purpose of this study was to evaluate the physical, chemical, and sensory changes in bran from three rice cultivars according to microwave roasting time. This study analyzed three rice cultivars, BRS Sertaneja (S), BRS Primavera (P), and IRGA 417 (I) determining the color parameters (L*, a*, and b *) at 6, 9, 12, 15, and 18 minutes of roasting time. After applying the difference from control test, the rice brans with different characteristics aroma and flavor were selected: S and P roasted for 9 and 15 minutes and IRGA 417 roasted for 9, 12, and 15 minutes. These samples were characterized by Free-Choice Profile descriptive sensory analysis, and their chemical composition was also determined. The longer the roasting process, the higher the roasted flavor intensity and aroma. The IRG 417 cultivar roasted for 12 minutes showed a sweeter flavor and aroma. After roasting, the brans remained rich in protein and lipid and presented higher fiber content and lower reducing sugar and phytic acid content. Microwave roasting for 12 minutes can be a viable option for improving the sensory functional and nutritional characteristics of the rice bran considering its use in food products.

Optimization of extrusion variables for the production of snacks from by-products of rice and soybean

This study aimed to define the process conditions to obtain snacks from the by-products of rice and soybean with physical characteristics suitable for marketing. Therefore, the effects of moisture and extrusion temperature on the expansion and color of the products obtained experimentally obtained were evaluated, and the proximate composition of the by-products and that of the snack with greater desirability were determined. Response surface methodology and rotational central composite design were used, and desirability test based on the regression models adjusted was applied. The most desirable snack, with the highest expansion index (3.39), specific volume (13.5 mL.g-1), and the chromaticity coordinate a* (2.79), was obtained under 12 g.100 g-1 moisture and 85ºC of temperature in the third zone of the extruder. The snack produced under these conditions attained content of protein and lipid content 41 and 64% higher than that of the traditional corn snack. It can be concluded that producing extruded snack made form a mixture of broken grains, rice bran, and soybean okara (81:9:10) is technologically feasible, enabling the development of a new product with good nutritional value that can improve the diet of children, the main consumers of this type of food.

Multivariate study and regression analysis of gluten-free granola

This study developed a gluten-free granola and evaluated it during storage with the application of multivariate and regression analysis of the sensory and instrumental parameters. The physicochemical, sensory, and nutritional characteristics of a product containing quinoa, amaranth and linseed were evaluated. The crude protein and lipid contents ranged from 97.49 and 122.72 g kg-1 of food, respectively. The polyunsaturated/saturated, and n-6:n-3 fatty acid ratios ranged from 2.82 and 2.59:1, respectively. Granola had the best alpha-linolenic acid content, nutritional indices in the lipid fraction, and mineral content. There were good hygienic and sanitary conditions during storage; probably due to the low water activity of the formulation, which contributed to inhibit microbial growth. The sensory attributes ranged from 'like very much' to 'like slightly', and the regression models were highly fitted and correlated during the storage period. A reduction in the sensory attribute levels and in the product physical stabilisation was verified by principal component analysis. The use of the affective test acceptance and instrumental analysis combined with statistical methods allowed us to obtain promising results about the characteristics of gluten-free granola.

Influence of the addition of lupine protein isolate on the protein and technological characteristics of dough and fresh bread with added Brea Gum

The effect of protein lupine isolate (LI) and addition of brea gum (BG) on a basic bread formulation is described. The major objective of this research was to evaluate the influence of the addition of LI on the quality and quantity of the proteins of fresh bread with BG. Protein quality was determinate by the Chemical Score method corrected for protein digestibility (CSCD%). The bread dough characteristics were determined by farinograph and alveograph. Fresh bread characterization was performed by measuring the physical parameters and evaluating the crumb structure. The effect of LI and BG on available lysine, the loss of available lysine ratio, and the chemical composition of the breads were also determined. The addition of LI on the bread formulation improved the protein content and the CSCD% of lysine. The dough with LI was less resistant to prolonged kneading and less manageable. With BG addition, the dough became stickier. The quality of fresh bread was affected by the addition of LI: the fresh bread had lower specific volume and more heterogeneous crumbs than that of the control group. The addition of BG did not influence the quality of the bread made with the mixed flour, but it had a positive effect on the loss of available lysine.

Carotenoids are related to the colour and lipid content of the pequi (Caryocar brasiliense Camb.) pulp from the Brazilian Savanna

This study investigated the colour, proximate composition, bioactive compounds (phenolic and carotenoid contents), and antioxidant activity of the pulp of pequi from different regions of the Brazilian Savanna. The colour parameters and the lipid and carotenoid contents of the pulp were significantly different between the samples. The lipid content ranged from 135.4 to 322.5 g/kg. The pequi pulp showed high total phenolic content (1.8 to 3.3 mg GAE/g). The carotenoid amount ranged from 37 to 187 µg/g. The carotenoid content was significantly correlated with the colour and lipid content of the pequi pulp. The antioxidant activity showed a mean IC50 value of 197.9 µg/mL. The pequi pulp is rich in phenolic compounds and carotenoids and has a good antioxidant activity. Its colour is influenced by the carotenoid content, which can be predicted by regression models using routine colour parameters.

Family rivalry in the testis: Retinoblastoma protein and E2F transcription factors in testicular development and adult spermatogenesis

Disorders of male reproductive health are becoming increasingly prevalent globally. These defects, ranging from decreasing sperm counts to an increasing rate of infertility and testicular cancer, have a common origin in the early phases of testicular development, but the exact mechanisms that cause them remain unknown. Testicular development and adult spermatogenesis are complex processes in which different cell types undergo mitosis, meiosis, differentiation and apoptosis. The retinoblastoma protein family and its associated E2F transcription factors are key regulators of these cellular events. In the present study, the functions of these factors in postnatal testicular development and adult spermatogenesis were explored using different animal models. In addition, a new application of flow cytometry to study testicular cell dynamics was developed. An ablation of retinoblastoma protein in mouse Sertoli cells resulted in their cell cycle re-entry in adult testes, dedifferentiation and a severe spermatogenic defect. We showed that deregulated E2F3 contributed to these changes. Our results indicated that the E2F1 transcription factor is critical for the control of apoptosis in the developing postnatal testis. In the adult testis, E2F1 controls the maintenance of the spermatogonial stem cell pool, in addition to inhibiting apoptosis of spermatocytes. In summary, this study elucidated the complex interdependencies of the RB and E2F transcription factor families in the control of postnatal testicular development and adult spermatogenesis. Furthermore, this study provided a new methodology for the analysis of testicular cells.

Host-parasite relations in a mycoparasitic system : alterations in membrane permeability and lipid composition in Choanephora cucurbitarum infected by Piptocephalis virginiana /

examined in Choanephora cucurbita rum during the early stages of infection by Piptocephalis virginiana » There was a small but consistent increase in the leakage of electrolytes, amino acids and sugars as a result of infection. These low levels of differential leakage in infected tissues are explained on the basis of the nature of this obligate, biotrophic, mycoparasitic system. Quantitative analysis of the twenty six amino acids and amino compounds detected in the leacheates — showed similar profiles in infected and control host and no new species of amino acids or amino compounds were detected in either infected or control host leacheates. Comparatively high amounts of aspartic acid, glutamic acid and alanine were found in the leacheates of host and infected host . Analyses of the sugars comprising the leacheates of infected and control host showed the presence of eight sugars, among which glucose was found in significant amounts (50-53%) ' The nutritional implication of this preferential leakage is discussed. No significant difference was observed in the leacheates of infected host sugar profiles compared with that of the control host. Profiles of the internal pool sugars of infected and control host did not reflect that obtained from the leacheate data, perhaps owing to leakage of sugars in a selective manner . Membrane lipid analyses yielded higher levels of lipid in infected host compared with the control, both at the 24 h and 36 h analyses. In addition, preliminary investigations of phosphorous-32 incorporation and turnover in phospholipids showed higher levels of 32p incorporation and turnover in infected host compared with the control. No apparent difference was noted in the profiles of the neutral lipid classes and the polar lipid classes of the membrane lipids as determined by one and two dimensional thin-layer chromatography respectively. However, a small but consistently higher degree of unsaturation was detected in the fatty acids of infected tissue compared with the control. Also, '^''-^^''^^'-'-^'^^c acid, a polyunsaturated fatty acid previously reported to show a direct correlation during the early stages of infection and the degree of parasitism of P. virginiana on C. cucurbitarum , was found in higher amounts in infected host membrane lipids compared with that of the control host. The implications of these membrane lipid alterations are discussed with particular reference to the small but consistently higher leakage of electrolytes, amino acids and sugars observed during infection in this study.

Effect of portacaval anastomosis on glutamine synthetase protein and gene expression in brain, liver and skeletal muscle.

The effects of chronic liver insufficiency resulting from end-to-side portacaval anastomosis (PCA) on glutamine synthetase (GS) activities, protein and gene expression were studied in brain, liver and skeletal muscle of male adult rats. Four weeks following PCA, activities of GS in cerebral cortex and cerebellum were reduced by 32\% and 37\% (p<0.05) respectively whereas GS activities in muscle were increased by 52\% (p<0.05). GS activities in liver were decreased by up to 90\% (p<0.01), a finding which undoubtedly reflects the loss of GS-rich perivenous hepatocytes following portal-systemic shunting. Immunoblotting techniques revealed no change in GS protein content of brain regions or muscle but a significant loss in liver of PCA rats. GS mRNA determined by semi-quantitative RT-PCR was also significantly decreased in the livers of PCA rats compared to sham-operated controls. These findings demonstrate that PCA results in a loss of GS gene expression in the liver and that brain does not show a compensatory induction of enzyme activity, rendering it particularly sensitive to increases in ammonia in chronic liver failure. The finding of a post-translational increase of GS in muscle following portacaval shunting suggests that, in chronic liver failure, muscle becomes the major organ responsible for the removal of excess blood-borne ammonia.

Utilization of biodiesel-derived glycerol or xylose for increased growth and lipid production by indigenous microalgae

The use of industrial wastes rich in mineral nutrients and carbon sources to increase the final microalgal biomass and lipid yield at a low cost is an important strategy to make algal biofuel technology viable. Using strains from the microalgal collection of the Université de Montréal, this report shows for the first time that microalgal strains can be grown on xylose, the major carbon source found in wastewater streams from pulp and paper industries, with an increase in growth rate of 2.8 fold in comparison to photoautotrophic growth, reaching up to µ=1.1/day. On glycerol, growth rates reached as high as µ=1.52/day. Lipid productivity increased up to 370% on glycerol and 180% on xylose for the strain LB1H10, showing the suitability of this strain for further development for biofuels production through mixotrophic cultivation.