Postharvest diseases in citrus and characterization of the fungal population in Sao Paulo`s wholesale market

The purposes of this workwere to characterize postharvest injuries and to evaluate the physicochemical characteristics of`Nra` and `Lima`oranges and `Murcott` tangor at Ceagesp market, as well as to characterize the environmental mycoflora in retail points at Ceagesp in 2006. Fruits collected at retail points were stored for 14 days at 25 degrees C and 85-90% RH. The incidence of injuries was visually evaluated every three days. The physicochemical characteristics analyzed were titratable acidity and soluble solids amount. The environmental mycoflora was sampled according to the gravimetric method, using Petri dishes containing potato-dextrose-agar medium+pentabiotic opened for two minutes. The average rot incidences in `Pera` and `Lima` oranges and `Murcott` tangor were 12.8, 14.9 and 25.8%, respectively, at the end of the storage period, and green mold was the main postharvest disease. Associations between physicochemical parameters and rot incidence was, in general, not significant. The environmental fungal population varied significantly between the sampling months in retail points with an average of 25.3 cfu/plate. Penicillium and Cladosporium were the most recorded genera of fungi. Positive correlation (r=0.96) was observed between frequency of P digitatum found in the environment of retail points and the green mold in on-sale fruits of `Pera` orange. However, for `Lima` orange and `Murcott` tangor such a correlation was not verified.

Air trapping: The major factor limiting diaphragm mobility in chronic obstructive pulmonary disease patients

Background and objective: Patients with COPD can have impaired diaphragm mechanics. A new method of assessing the mobility of the diaphragm, using ultrasound, has recently been validated. This study evaluated the relationship between pulmonary function and diaphragm mobility, as well as that between respiratory muscle strength and diaphragm mobility, in COPD patients. Methods: COPD patients with pulmonary hyperinflation (n = 54) and healthy subjects (n = 20) were studied. Patients were tested for pulmonary function, maximal respiratory pressures and diaphragm mobility using ultrasound to measure the craniocaudal displacement of the left branch of the portal vein. Results: COPD patients had less diaphragm mobility than did healthy individuals (36.5 +/- 10.9 mm vs 46.3 +/- 9.5 mm, P = 0.001). In COPD patients, diaphragm mobility correlated strongly with pulmonary function parameters that quantify air trapping (RV: r = -0.60, P < 0.001; RV/TLC: r = -0.76, P < 0.001), moderately with airway obstruction (FEV1: r = 0.55, P < 0.001; airway resistance: r = -0.32, P = 0.02) and weakly with pulmonary hyperinflation (TLC: r = -0.28, P = 0.04). No relationship was observed between diaphragm mobility and respiratory muscle strength (maximal inspiratory pressure: r = -0.11, P = 0.43; maximal expiratory pressure: r = 0.03, P = 0.80). Conclusion: The results of this study suggest that the reduction in diaphragm mobility in COPD patients is mainly due to air trapping and is not influenced by respiratory muscle strength or pulmonary hyperinflation.

Factors affecting fertilisation and early embryo quality in single- and superovulated dairy cattle

Data on fertilisation and embryo quality in dairy cattle are presented and the main factors responsible for the low fertility of single-ovulating lactating cows and embryo yield in superovulated dairy cattle are highlighted. During the past 50 years, the fertility in high-producing lactating dairy cattle has decreased as milk production increased. Recent data show conception rates to first service to be approximately 32% in lactating cows, whereas in heifers it has remained above 50%. Fertilisation does not seem to be the principal factor responsible for the low fertility in single-ovulating cows, because it has remained above 80%. Conversely, early embryonic development is impaired in high-producing dairy cows, as observed by most embryonic losses occurring during the first week after fertilisation. However, in superovulated dairy cattle, although fertilisation failure is more pronounced, averaging approximately 45%, the percentage of fertilised embryos viable at 1 week is quite high (>70%). Among the multifactorial causes of low fertility in lactating dairy cows, high feed intake associated with low concentrations of circulating steroids may contribute substantially to reduced embryo quality. Fertilisation failure in superovulated cattle may be a consequence of inappropriate gamete transport due to hormonal imbalances.

Fatty acid profile and qualitative characteristics of meat from zebu steers fed with different oilseeds

The objective of this study was to evaluate the effects of dietary addition of ground oilseed sources on the quality, fatty acid profile, and CLA content of meat from zebu steers. Thirty-one zebu steers with an initial average age of 23 mo and an initial BW of 365 kg were used in this study. The experimental period was 84 d, which was preceded by an adaption period of 28 d. The diet was provided ad libitum with a forage: concentrate ratio of 40:60. Corn silage was used as the forage source. Four different concentrates were formulated for each treatment: without additional lipids (control) or with ground soybeans (SB), ground cottonseed (CS), or ground linseed (LS). The SB, CS, and LS diets were formulated to have 6.5% ether extract on a total dietary DM basis. The experiment was set up as a completely randomized design. After slaughter, samples were taken from the longissimus thoracis muscle for the measurement of fatty acid concentration and the evaluation of meat quality. The luminosity index was greater in the control and LS diets (P < 0.01). The greatest percentages of myristic acid (C14:0), palmitic acid (C16:0), trans octadecenoic acid (C18:1 trans-10, trans-11, or trans-12), and SFA in the subcutaneous fat were observed in the CS treatment (P < 0.01). Moreover, the least percentages of oleic acid (C18:1 cis-9) and total unsaturated fatty acids in the subcutaneous fat were observed in the CS diet (P < 0.01). The meat linoleic acid and a-linolenic acid percentages were greatest in the SB and LS treatments, respectively (P < 0.001). The unsaturated fatty acid: SFA ratio was smallest for the CS diet (P < 0.01). A gradual increase in oxidation was observed as a function of storage time; however, the diets did not affect the rancidity of the meat (P > 0.05). The fatty acid profile of subcutaneous fat was impaired by the addition of CS. Supplying ground oilseeds did not increase the content of CLA in the meat.

Milk yield, milk composition, eating behavior, and lamb performance of ewes fed diets containing soybean hulls replacing coastcross (Cynodon species) hay

The objective of this experiment was to evaluate the effects of replacing coastcross hay NDF by soybean hull (SH) NDF on the lactation performance and eating behavior of ewes and also on the performance of their lambs. Fifty-six Santa Ines lactating ewes (56.1 +/- 6.8 kg of initial BW; mean +/- SD) were penned individually and used in a randomized complete block design with 14 blocks and 4 treatments. Diets were formulated to provide similar concentrations of NDF (56%) and CP (16%). The SH NDF replaced 33 (SH33), 67 (SH67), or 100% (SH100) of the NDF contributed by coastcross hay in a 70% forage-based diet (SH0), resulting in SH inclusion rates of 0, 25, 54, and 85% of the dietary DM. Once a week, from the second to the eighth week of lactation (weaning time), ewes were separated from their lambs, stimulated by a 6-IU i.v. oxytocin injection, and hand milked to empty the udder. After 3 h, milk production was obtained after the same procedure. Quadratic effect for milk production (142.4, 179.8, 212.6, and 202.9 g/3 h) and cubic effect for DMI (2.27, 2.69, 3.25, and 3.00 kg/d) were observed as SH inclusion increased from 0 to 85% of the dietary DM. Milk fat (7.59, 7.86, 7.59, and 7.74%), protein (4.53, 4.43, 4.40, and 4.55%), and total solids (18.24, 18.54, 18.39, and 18.64%) did not differ among the 70% forage-based diet and diets with SH NDF replacing 33, 67, or 100% of the NDF. A linear increase in lactose concentration was observed with SH inclusion. Ewe BW gain during the trial showed a cubic response (0.37, 0.03, 4.80, and 2.80 kg) with SH inclusion. The preweaning ADG of lambs increased linearly, and ADG of lambs after weaning decreased linearly with SH inclusion. Final BW of lambs (2 wk after weaning) did not differ among treatments. Eating behavior observations were conducted with 44 ewes. The same facilities, experimental design, dietary treatments, and feeding management were used. Observations were visually recorded every 5 min for a 24-h period when ewes were 46 +/- 6.8 d in milk. Eating time (min/d, min/g of DMI, and min/g of NDF intake) and time expended in rumination and chewing activities (min/g of DMI and min/g of NDF intake) decreased linearly with the addition of SH in the diets. The inclusion of SH improved DMI and milk production, also reflecting on the BW of lambs at weaning. Milk performance was not affected when SH NDF replaced 100% of hay NDF.

Feeding Dietary Mannan Oligosaccharides to Juvenile Nile Tilapis, Oreochromis niloticus, Has No Effect on Hematological Parameters and Showed Decreased Feed Consumption

Impaired immune system by environmental stressors can lead fishes to be more susceptible to diseases that limit the economic development of aquaculture systems. This study was set out to determine the effect of six levels of mannan oligosaccharides (MOS; ActiveMOS((R)); Biorigin, Lencois Paulista, Sao Paulo, Brazil) on the performance index and hematology of Nile tilapia, Oreochromis niloticus juveniles. Fish (13.62 g) were randomly distributed into 18 plastic aquaria (300 L; 20 fishes per aquarium) and fed during 45 d with a commercial diet supplemented with 0, 0.2, 0.4, 0.6, 0.8, and 1% dietary MOS, in a totally randomized design trial (n = 3); biometrical and hematological data were collected and analyzed. There were no significant differences in hematological parameters between fish fed control and MOS supplementation diets, and daily feed consumption (FC) decreased (P < 0.05) with increasing levels of dietary MOS. Dietary MOS did not increase leukocyte count and presented negative effects on FC of Nile tilapia. At 0.4% MOS supplementation, the individual weight gain was higher in absolute values but not different (P > 0.05) compared to control diet.

Brain nitric oxide production by a proline-rich decapeptide from Bothrops jararaca venom improves baroreflex sensitivity of spontaneously hypertensive rats

Baroreflex sensitivity is disturbed in many people with cardiovascular diseases such as hypertension. Brain deficiency of nitric oxide (NO), which is synthesized by NO synthase (NOS) in the citrulline-NO cycle (with argininosuccinate synthase (ASS) activity being the rate-limiting step), contributes to impaired baroreflex. We recently showed that a decapeptide isolated from Bothrops jararaca snake venom, denoted Bj-PRO-10c, exerts powerful and sustained antihypertensive activity. Bj-PRO-10c promoted vasodilatation dependent on the positive modulation of ASS activity and NO production in the endothelium, and also acted on the central nervous system, inducing the release of GABA and glutamate, two important neurotransmitters in the regulation of autonomic systems. We evaluated baroreflex function using the regression line obtained by the best-fit points of measured heart rate (HR) and mean arterial pressure (MAP) data from spontaneously hypertensive rats (SHRs) treated with Bj-PRO-10c. We also investigated molecular mechanisms involved in this effect, both in vitro and in vivo. Bj-PRO-10c mediated an increase in baroreflex sensitivity and a decrease in MAP and HR. The effects exerted by the peptide include an increase in the gene expression of endothelial NOS and ASS. Bj-PRO-10c-induced NO production depended on intracellular calcium fluxes and the activation of a G(i/o)-protein-coupled metabotropic receptor. Bj-PRO-10c induced NO production and the gene expression of ASS and endothelial NOS in the brains of SHRs, thereby improving baroreflex sensitivity. Bj-PRO-10c may reveal novel approaches for treating diseases with impaired baroreflex function. Hypertension Research (2010) 33, 1283-1288; doi: 10.1038/hr.2010.208

Hemoglobin Regeneration Efficiency in Anemic Rats: Effects on Bone Mineral Composition and Biomechanical Properties

This study reports the effects of dietary iron (Fe) deficiency and recovery on bone mineral composition and strength in anemic rats submitted to a hemoglobin (Hb) repletion assay. Weanling male Wistar rats were fed a low-Fe diet (12 mg/kg) for 15 days followed by 2 weeks of Fe repletion with diets providing 35 mg Fe/kg as either ferrous sulfate (n = 8) or ferric pyrophosphate (FP; n = 12). At final day of each period (depletion and repletion), Fe-adequate animals were also euthanized. Iron status (blood Hb, Hb Fe pool, Hb regeneration efficiency), tibia mineral concentrations (Ca, Mg, Fe, Cu, and Zn) and biomechanical properties were evaluated. Iron-deficient rats had lower tibia Fe and Mg levels and bone strength when compared to controls. Yield load and resilience were positively related to tibia Mg levels (r = 0.47, P = 0.02 and r = 0.56, P = 0.004, respectively). Iron repletion did not recover tibia Mg concentrations impaired by Fe deficiency. Moreover, bone elastic properties were negatively affected by FP consumption. In conclusion, bone mineral composition and strength were affected by Fe deficiency, whereas dietary Fe source influenced tibia Mg and resistance in the period during which rats were recovering from anemia.

Changes in bone mass, biomechanical properties, and microarchitecture of calcium- and iron-deficient rats fed diets supplemented with inulin-type fructans

Feeding mineral-deficient diets enhances absorptive efficiency as an attempt of the body to compensate for the lack of an essential nutrient. Under certain circumstances, it does not succeed, and nutritional deficiency is produced Our hypothesis was that mulin-type fructans (ITF), which arc known to affect mineral absorption, could increase Ca and Fe bioavailability in Ca- and Fe-deficient rats. Male Wistar rats (n = 48, 4 weeks old) were assigned to I of 8 groups derived from 2 x 2 x 2 factorial design with 2 levels of added Fe (0 and 35 mg/kg), Ca (0 and 5 g/kg), and ITF (0 and 100 g/kg) for 33 days. The Fe status (hemoglobin, serum Fe, total Fe-binding capacity, transferrin saturation, liver minerals) was evaluated. Tibia minerals (Ca, Mg, and Zn), bone strength, and histomorphometry were determined In nondeficient rats, ITF supplementation did not affect Fe status or organ minerals, with the exception of tibia Mg Moreover, ITF improved bone resilience and led to a reduction in eroded surface per body surface and number of osteoclasts per area In Ca-deficient rats, ITF increased liver (Fe and Zn) and tibia (Zn) mineral levels but impaired tibia Mg, yield load, and resilience. In conclusion, ITF worsened the tibia Mg levels and elastic properties when supplemented in Ca-deficient diets In contrast, although bone Ca was not affected in nondeficient rats under the present experimental conditions, bone quality improved, as demonstrated by a moderate reduction in femur osteoclast resorption and significant increases in tibia Mg content and elasticity. (C) 2009 Elsevier Inc. All rights reserved.

Chlorophyll Degradation and Formation of Colorless Chlorophyll Derivatives during Soybean (Glycine max L. Merill) Seed Maturation

The natural chlorophyll degradation results in noncolored chlorophyll catabolites (NCCs), but there are controversies if these are the final products. The formation and degradation of NCCs during soybean seed (Glycine max L. Merrill) maturation and two drying temperatures were investigated. Soybean was harvested at six maturation stages. The effect of postharvest drying at 40 and 60 degrees C on the NCC formation was analyzed by high-performance liquid chromatography (HPLC), and results were expressed as areas under the curve. All samples contained fractions with an absorption maximum at 320 nm, typical for NCC. The amounts of NCC increased until 114 days after planting and were significantly lower in advanced maturation stages. These results indicate that the NCC in soybeans might not be the final products of chlorophyll degradation. Their reduction in advanced maturation stages may be due to further metabolization. Heating soybeans at 40 and 60 degrees C promoted unnatural chlorophyll degradation and impaired the formation of NCC.

Detection of the TLR4 1196C > T Polymorphism by Mismatched-Polymerase Chain Reaction Using Plasmid DNA as Internal Control in Restriction Fragment Length Polymorphism Assays

Background: Restriction fragment length polymorphism (RFLP) is a common molecular assay used for genotyping, and it requires validated quality control procedures to prevent mistyping caused by impaired endonuclease activity. We have evaluated the usefulness of a plasmid-based internal control in RFLP assays. Results: Blood samples were collected from 102 individuals with acute myocardial infarction (AMI) and 108 non-AMI individuals (controls) for DNA extraction and laboratory analyses. The 1196C> T polymorphism in the toll-like receptor 4 (TLR4) gene was amplified by mismatched-polymerase chain reaction (PCR). Amplicons and pBluescript II SK-plasmid were simultaneously digested with endonuclease HincII. Fragments were separated on 2% agarose gels. Plasmid was completely digested using up to 55.2 nmL/L DNA solutions and 1 mu L PCR product. Nevertheless, plasmid DNA with 41.4 nM or higher concentrations was incompletely digested in the presence of 7 mL PCR product. In standardized conditions, TLR4 1196C> T variant was accurately genotyped. TLR4 1196T allele frequency was similar between AMI (3.1%) and controls (2.0%, p = 0.948). TLR4 SNP was not associated with AMI in this sample population. In conclusion, the plasmid-based control is a useful approach to prevent mistyping in RFLP assays, and it is validate for genetic association studies such as TLR4 1196C> T.

Lopap: A non-inflammatory and cytoprotective molecule in neutrophils and endothelial cells

Lopap (Lonomia obliqua prothrombin activator protease) is a member of the lipocalin family isolated from the extract of L obliqua bristles. Lopap displays serine protease-like activities, including coagulation disturbance, cytokine secretion and antiapoptotic activity in human cultured endothelial cells. Here, we have investigated the effects of the recombinant protein (rLopap) on the inflammatory and apoptotic processes of neutrophils and endothelial cells from male Wistar rats. We found that rLopap did not induce in vivo leukocyte-endothelial interactions in the microvasculature, initial steps of leukocyte recruitment during inflammation. Incubation of rLopap with neutrophils or endothelial cells prevented apoptosis evoked by serum deprivation and induced nitric oxide (NO) production in both cell types, and increased the expression of ICAM-1 by endothelial cells. Simultaneous incubation of endothelial cells or neutrophils with rLopap and N(omega)-nitro-L-arginine methyl ester (L-NAME), a non-specific inhibitor of NO synthases, inhibited NO production and impaired the protection on apoptosis. Differently, incubation of endothelial cells with monoclonal antibody anti ICAM-1 did not change the protection on apoptosis evoked by rLopap. Together, these results indicate that rLopap does not display inflammatory properties in vivo but inhibits apoptosis of neutrophils and endothelial cells depending, at least in part, on NO production. (C) 2009 Elsevier Ltd. All rights reserved.

Biomarkers of oxidative stress and endothelial dysfunction in glucose intolerance and diabetes mellitus

Objectives: To evaluate biomarkers of endothelial dysfunction and oxidative stress in glucose intolerance (GI) compared to overt diabetes (DM2). Design and methods: 140 volunteers including 96 with DM2, 32 with GI and 12 controls (C) were Studied. NO metabolites, NO synthase inhibitors. thiols and N-acetyl-beta-glucosaminidase (NAGase) activity were analyzed by chemiluminescence, capillary electrophoresis, ELISA and colorimetric assay, respectively. Results: (center dot)NO metabolites were higher in GI (NOx: P=0.03 S-nitrosothiols: p=0.001) and DM2 (p=0.006; p=0.0006) groups in relation to group C, while nitrotyrosine was higher only in the DM2 group in comparison 10 the other groups. NAGase activity was elevated in GI (p=0.003) and DM2 (p=0.0004) groups in relation to group C, as well as, ADMA (p=0.01: p=0.003) and GSSG (p=0.01 p=0.002). Conclusions: (center dot)NO metabolites. (center dot)NO synthase inhibitors. thiols and NAGase are biomarkers Suitable to indicate endothelial dysfunction and oxidative stress in the early stages of impaired response to insulin. (c) 2008 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Gastro-resistant Pellets of Didanosine obtained by Extrusion and Spheronization: Assessing the Production Process

The aim this work was develop gastro-resistant pellets of didanosine as well as study the impact on the pellets properties, regarding the way as the binder was added and drying process used. The pellets formation was accompanied by analysis of morphological parameters and didanosine dissolution. In the most cases, pellets showed diameter around 1.0 mm and shape parameters acceptable. The variations of the process did not interfere significantly in pellets size. In turn, drying in fluid bed favored the dissolution of didanosine, in contrast to binder addition on powder form that impaired. In another hand, this last resulted in the best aspect factor (about 1.1). Gastro-resistant pellets showed adequate dissolution, compatible with this type of dosage form. The variables of process studied enabled obtain pellets with characteristics of shape and dissolution just slightly different, indicating flexibility of the formulation for production of gastro-resistant pellets of didanosine.

Aortas isolated from sinoaortic-denervated rats exhibit rhythmic contractions that are regulated by pharmacologically distinct calcium sources

Sinoaortic denervation is characterized by arterial pressure lability, without sustained hypertension. Aortas isolated from rats with sinoaortic denervation present rhythmic contractions. We studied the participation of distinct Ca2+ sources in the maintenance of the oscillations. Three days after the surgeries, aortic rings were placed in an organ chamber, and the incidence of aortas presenting rhythmic contractions was measured. Specific drugs were employed to analyse the participation of the Ca2+ released from the sarcoplasmic reticulum [2-APB (diphenylborinic acid 2-aminoethyl ester), thapsigargin and ryanodine] and external Ca2+ entry [Bay K 8644, verapamil and DMB (dimethylbenzyl amiloride)] on the rhythmic contractions. Additionally, we verified the effects of chloride channel blocker NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] on the maintenance of the rhythmic contractions. Under phenylephrine stimulus, sinoaortic-denervated rat aortas exhibited rhythmic contractions in the frequency of 4.5 +/- 0.50 cycles/min. and an amplitude of 0.465 +/- 0.05 g. 2-APB, thapsigargin and ryanodine inhibited the rhythmic contractions. Bay K 8644 increased the oscillations, reaching maximum values with a concentration of 50 nM (18.5 +/- 2.5 cycles/min.). The rhythmic contractions were inhibiting by verapamil and Ca2+-free solution. DMB and NPPB did not alter the oscillations. In conclusion, we observed that aorta isolated from sinoaortic-denervated rats present rhythmic contractions. Moreover, drugs that impaired intracellular Ca2+ release from sarcoplasmic reticulum interrupted the oscillations. The oscillations also depend on the extracellular Ca2+ entry through L-type Ca2+.