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Ventilator-associated pneumonia (VAP) has been described in humans and in experimental animals. The most severe lesions are located in dependent lung segments along a sterno-vertebral axis, however the cephalocaudal distribution of lung infection remains unknown. We used an experimental model to evaluate the distribution of lung infection, considering its anteroposterior and cephalocaudal gradient, and its impact on lung aeration. Ten healthy domestic piglets were anesthetized, paralyzed and mechanically ventilated for 59 hours in the prone position. At the end of the experiment they were sacrificed and their lungs were fixed. Six segments were analyzed: a non-dependant (ND) and a dependant (D) segment of the upper (UL), middle (ML) and lower (LL) lobes. The presence of healthy lung or of histological infectious lesions was analyzed with a semi-quantitative method. The regional distribution of lung infection was compared between upper, middle and lower lobes, as well as between dependant and non-dependant regions. The presence of infectious lesions was correlated with measurements of lung aeration. Nine of the ten piglets developed VAP. Infectious lesions were distributed along a sterno-vertebral and a cephalocaudal gradient; the lower and middle lobes were more frequently infected than the upper lobes. There was an inverse correlation (R= - 0.902) between the development of lung lesions and lung aeration. In conclusion, VAP was a frequent complication in healthy mechanically ventilated piglets, showing an anteroposterior as well as a cephalocaudal gradient. As expected, development of lung infection was accompanied by a corresponding loss of aeration.

The high incidence of Hepatitis A and B in institutionalized patients with Down Syndrome (DS) is not fully understood. Under poor hygienic conditions, immunological alterations might predispose individuals to these infections. Sixty three DS children between 1 and 12 years old living at home with their families were examined for anti-HAV and compared to age-matched controls (64 healthy children). This cross-sectional study was carried out from May, 1999, to April, 2000, at the Hospital de Clínicas of Porto Alegre, southern Brazil. Groups were compared in terms of age, sex, skin color, and family income (> R$ 500 and < R$ 500/ month) by the chi-square test, with Yates' correction and for the prevalence of anti-HAV (Fisher's exact test). In the DS group (n=63), the mean age was 4.4 ± 3.3 years, 94% of the patients were white and 51% were female. Family income was

Leptospirosis may have important complications, such as acute respiratory failure (ARF) associated or not with other organic dysfunction, with a high mortality rate. We report the characteristics and evolution of severe leptospirosis associated with ARF. During 10 years, 35 consecutive adult patients admitted in two general Intensive Care Units with severe leptospirosis and ARF, were followed up. Clinical characteristics, associated organic dysfunction and mortality were analyzed. Survivors were compared with non-survivors. The most frequent clinical manifestations were dyspnea, fever, myalgia, jaundice, hemoptysis and coughing. All patients presented ARF, needing mechanical ventilation, as well as other organic dysfunctions. The mortality rate was 51%. Non-survivors were older and had a higher incidence of organic dysfunction, mainly renal, cardiovascular and neurological failures, as well as a higher level of acidosis. In conclusion, leptospirosis should be considered as a cause of severe ARF and other associated organic dysfunctions.

Objetivo: avaliar a prevalência de hepatite viral A (HVA) em crianças e adolescentes portadores de doenças crônicas do fígado, em um serviço de hepatologia pediátrica. Métodos: entre maio de 1999 e fevereiro de 2001, foi estudada a prevalência de anticorpos anti-HVA total em 60 crianças e adolescentes, entre 1 e 16 anos de idade, portadoras de hepatopatias crônicas, provenientes da unidade de gastroenterologia pediátrica e programa de transplante hepático infantil do serviço de pediatria do Hospital de Clínicas de Porto Alegre. O anti-HVA, realizado através de um teste laboratorial comercialmente disponível em nosso meio (Abbott - MEIA HAVAB - sistema AXSYM), foi determinado e relacionado com a idade, com o sexo, com a cor, com o diagnóstico etiológico da hepatopatia e com a renda familiar dos pacientes. Resultados: apenas uma criança de 1 ano, portadora de atresia biliar, foi excluída do estudo por apresentar anti-HVA indeterminado, em duas ocasiões. Das 59 crianças restantes, 14 (24%) apresentavam resultados positivos para o anti-HVA total. As idades dos pacientes com anti-HVA positivos variaram de 1 a 16 anos (x= 7,7 anos e mediana 8,5 anos). Não houve diferença significante entre idade, sexo e cor entre os grupos positivo e negativo. A renda familiar foi menor no grupo dos pacientes anti-HVA positivo, mas não mostrou diferença estatística significante. A diferença de prevalência de anti-HVA entre as etiologias das hepatopatias está, provavelmente, relacionada à idade mais do que ao diagnóstico. Conclusões: na população estudada, a maioria (76%) das crianças e adolescentes portadoras de hepatopatias crônicas é suscetível à infecção pelo vírus A, podendo apresentar, portanto, um curso mais grave e complicações, se adquirirem HVA. Sugerimos, então, que esses pacientes devam receber a vacina inativada contra HVA.

We present and discuss in this work specific-heat experiments at low temperature that we have conducted on different molecular (hydrogenbonded) glasses and crystals. Specifically, we have measured the low-temperature specific heat Cp(T) for a set of solid alcohols: normal and fully-deuterated ethanol, 1−and 2−propanol, and glycerol. Ethanol exhibits a very interesting polymorphism presenting three different solid phases at low temperature: a fully-ordered (monoclinic) crystal, an orientationally-disordered (cubic) crystal or “orientational glass”, and the canonical (amorphous) structural glass. By measuring and comparing the low-temperature specific heat of the three phases, in the boson peak range, 2−10 K, as well as in the tunneling-states range, below 1K, we are able to provide a quantitative confirmation that “glassy behavior” is not an exclusive property of amorphous solids. On the other hand, propanol is the simplest monoalcohol with two different stereoisomers (1−and 2−propanol), which allows us to study directly the influence of the spatial rearrangement of atoms on the universal properties of glasses. We have measured the specific heat of both isomers, finding a noteworthy quantitative difference between them. Finally, low-temperature specific-heat data of glassy and crystalline glycerol have also been obtained.En este trabajo, presentamos y discutimos experimentos de calorimetría a bajas temperaturas que hemos realizado en diferentes vidrios y cristales moleculares (de puentes de hidrógeno). En concreto, hemos medido el calor específico a bajas temperaturas Cp(T) para un conjunto de alcoholes sólidos: etanol normal y completamente deuterado, 1−y 2−propanol, y glicerol. El etanol presenta un muy interesante polimorfismo, con tres fases sólidas diferentes a bajas temperaturas: un cristal estable perfectamente ordenado (monoclínico), un cristal (cúbico) con desorden orientacional o “vidrio orientacional”, y el vidrio estructural canónico (amorfo). Midiendo y comparando el calor específico a bajas temperaturas de las tres fases, en el rango de temperaturas del pico bosónico (2−10 K) así como en el de los estados de tuneleo por debajo de 1K, hemos podido proporcionar una confirmación cuantitativa de que el “comportamiento vítreo” no es una propiedad exclusiva de los sólidos amorfos. Por otra parte, el propanol es el monoalcohol más sencillo que cuenta con dos estereoisómeros (1−y 2−propanol), lo que nos permite estudiar directamente la influencia de la organización espacial de los átomos en las propiedades universales de los vidrios. Hemos medido el calor específico de ambos isómeros, encontrando una notable diferencia cuantitativa entre ellos. Finalmente, también hemos obtenido el calor específico a bajas temperaturas del glicerol en sus fases vítrea y cristalina.

Sea turtles, with their global distribution and complex life cycle, often accumulate pollutants such as metals and metalloids due to their extended lifespan and feeding habits. However, there are limited studies exploring the impact of metal pollution on the reproductive health of female sea turtles, specifically focusing on the quality of their eggs, which has significant implications for the future generations of these charismatic animals. São Tomé Island, a crucial nesting and feeding habitat for green sea turtles, underscores the urgent need for comprehensive research in this ecologically significant area. This study aimed to investigate whether metals and metalloids in the blood of nesting female green sea turtles induce genotoxic effects in their erythrocytes and affect their egg morphometric characteristics and the composition of related compartments. Additionally, this study aimed to evaluate whether the quality of energetic reserves for embryo development (fatty acids in yolk's polar and neutral lipids) is influenced by the contamination status of their predecessors. Results revealed correlations between Cu and Hg levels and increased "lobed" erythrocytes, while As and Cu negatively influenced shell thickness. In terms of energy reserves, both polar and neutral lipid fractions contained primarily saturated and monounsaturated fatty acids, with prevalent 18:1n-9, 18:0, 16:0, 14:0, and 12:0 fatty acids in yolk samples. The yolk polar fraction was more susceptible to contaminant levels in female sea turtles, showing consistent negative correlations between pollution load index and essential n3 fatty acids, including linolenic, eicosatrienoic, eicosapentaenoic, and docosapentaenoic acids, crucial for embryonic development. These metals accumulation, coupled with the reduced availability of these key fatty acids, may disrupt the eicosanoid and other important pathways, affecting reproductive development. This study reveals a negative correlation between metal contamination in female sea turtles' blood and egg lipid reserves, raising concerns about embryonic development and the species' future generations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Iron is routinely supplemented in broiler feeds aiming to prevent dietary deficiencies. Limestone and phosphates are very rich in Fe; however, its contribution from these sources have not been thoroughly investigated with chickens. The present research was conducted to evaluate live performance and blood parameters of broilers when using limestone and dicalcium phosphate as sources of Fe. A total of 576 one-day-old male Cobb x Cobb 500 were allocated into a total of 72 battery cages, 6 treatments with 12 replication cages of 8 chicks at placement. Chicks were fed diets formulated with corn, soybean meal (SBM) with laboratory grade calcium carbonate and phosphoric acid (having traces of Fe). All chicks were fed a common prestarter without Fe supplementation (analyzed total 58.2 ± 2.4 mg/kg Fe) from placement to 7 d. Allocation of birds to dietary treatments was completely randomized on day 8. Treatments had increasing Fe derived from commercial limestone and dicalcium phosphate (analyzed Fe 7,218 and 4,783 mg/kg, respectively) progressively replacing calcium carbonate and phosphoric acid to provide graded increases in total Fe (analyzed Fe in the feeds were 57.6 ± 2.1, 92.0 ± 2.3, 124.1 ± 2.7, 159.3 ± 3.1, 187.2 ± 3.2, 223.7 ± 3.6 mg/kg, respectively). There were no effects of dietary Fe on live performance, hematocrit, and hemoglobin the end of the study on day 28 (P > 0.05). Increasing dietary Fe from commercial limestone and dicalcium phosphate led to a linear reduction in the percent ileal digestible Fe. However, linear increments in Fe retention, serum ferritin and liver Fe occurred when compared to feeds without Fe derived from limestone and phosphate dicalcium. It is concluded that Fe from limestone and dicalcium phosphate can be partially utilized by broiler chickens. It was estimated that the Fe retained from limestone and dicalcium phosphate is of 1.9%. Broilers fed corn-soy feeds (58.2 mg/kg Fe) do not require supplemental Fe., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Breeding snakes in captivity has become more and more relevant due not only to the growing interest on their venoms but also to the increasing number of endangered species worldwide. Unfortunately, studies on the formation of germplasm banks for these reptiles do not follow the same pace, and literature on sperm cryopreservation remains in its infancy when compared to other taxa. Herein, we first validated a sperm-egg binding assay (using chicken egg perivitelline membrane - EPM) and some nonfluorescent staining techniques for semen analysis of two pit viper genera (Bothrops and Crotalus), and then we investigated the protective effects of dimethylacetamide (DMA), dimethylformamide (DMF), and dimethylsulfoxide (DMSO) at different concentrations (3, 6 and 12%) throughout the freezing process in five species of lancehead and one of rattlesnake. Our validation process showed high correlations among sperm functional tests (including sperm-binding to EPM) and motion parameters. A total of 166 fresh ejaculates were acquired from 233 collection attempts, and 63.9% of these samples exhibited minimal motility for freezing (≥20%). During cryopreservation we observed that post-thaw motility and quality was improved by higher levels of cryoprotectants (CPA), regardless the CPA type. Lower concentrations of CPA were less harmful to sperm motility and progressive motility following the equilibrium phase, but were ineffective in protecting these cells from the freeze-thaw cycle. Likewise, higher CPA concentrations increased post-thaw integrity of the acrosome and plasma membrane for most species, except for rattlesnakes in which only 12% DMSO produced better outcomes., (Copyright © 2022. Published by Elsevier Inc.)

Wooden breast (WB) myopathy was investigated in broilers fed varying energy and protein at early ages. Correlation analyses were conducted between echogenicity of ultrasound images (US) of breast muscle from live birds and WB after slaughter. A total of 1,000 Cobb 500 one-day-old male chicks were fed on five dietary programs with eight replicates of 25 birds each, in a completely randomized design. Control feeds (commercially used ME and ideally balanced amino acids) or low-density feeds (low EP, with reductions of 50 kcal/kg ME and 0.20% dig. Lys compared to the control) were formulated. Feeds were provided in different periods: 1 to 7 d, 8 to 14 d, 15 to 21 d or 22 to 28 d. All broilers were fed a common basal diet thereafter until 49 d. Images using US were obtained once a week from all individuals and WB scored from one slaughtered bird per replication (0, normal; 1, mild hardening in the upper breast muscle; 2; moderate hardening in the upper and/or lower breast muscle; 3, severe hardening; 4, severe hardening with hemorrhagic lesions and yellow fluid). Blood was collected for enzyme investigation from the weekly slaughtered bird. Broilers had lower BWG and higher FCR when fed low EP feeds, regardless of the period fed when compared to the control (P < 0.001). Growth compensation, however, occurred afterwards such that all birds presented similar performance at the end. At 14, 21, and 28 d, broilers previously fed low EP feeds had lower WB scores (P < 0.001) compared to birds fed the control; however, both groups presented increased WB scores after 28 d. Wooden breast was positively correlated with breast echogenicity at 21 d (r = 0.31), 28 d (r = 0.43), 35 d (r = 0.21) and 42 d (r = 0.39). In conclusion, dietary energy and protein affected the development of WB scores in broilers and breast US images can be used as an early predictor of WB., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The present research was conducted to assess Mn requirements of broiler breeder hens. One hundred and twenty Cobb 500 hens, 22 wk of age, were individually allocated in cages. After fed a Mn-deficient diet (22.2 ppm), hens were randomly placed in treatments having 6 increments of 30-ppm Mn. All trace minerals were from laboratory grade sources being Mn from Mn sulfate (MnSO 4 H 2 O). Treatments were fed for 4 periods of 28 d. There were no interactions between dietary Mn and period for any evaluated response (P > 0.05). Requirements of Mn for hen day egg production and settable egg production were 115.8 and 56.6 ppm and 122.1 and 63.6 ppm (P < 0.05), respectively, using quadratic polynomial (QP) and broken line quadratic (BLQ) models, whereas total eggs and total settable eggs per hen had Mn requirements estimated at 115.7 and 56.6 and 121.8 and 61.7 ppm (P < 0.05), respectively. Number of cracked, defective, and contaminated eggs decreased, whereas hatchability, hatchability of fertile eggs, eggshell percentage, and eggshell palisade layer increased when hens were fed diets having 48.5 to 168.2-ppm Mn (P < 0.05). Maximum responses for egg weight and eggshell percentage were 117.7 and 63.6 ppm as well as 131.6 and 71.0 ppm (P < 0.05), respectively, using QP and BLQ models. Breaking strength and egg specific gravity had Mn requirements estimated at 140.2 and 112.7 ppm as well as 131.3 68.5 ppm (P < 0.05), whereas eggshell palisade layer and eggshell thickness were maximized with 128.8 and 68.8 ppm and 140.2 134.2 ppm, respectively, for QP and BLQ models (P < 0.05). Maximum yolk Mn content values were obtained using 118.0- and 118.4-ppm Mn by QP and BLQ models, respectively. The average Mn requirements estimated for QP and BLQ models is 128.4 and 92.3 ppm Mn (18.7 and 13.5 mg/hen/d), respectively, which is much lower than what has been currently recommended in commercial production., (Copyright © 2020. Published by Elsevier Inc.)

A study was conducted to evaluate the effects of a supplemental α-amylase on energy and nutrient utilization of broiler chickens fed diets with variable amounts of corn. A total of 480 slow feathering Cobb × Cobb 500 male broilers were randomly distributed into 10 treatments with 8 replicates of 6 birds each. Birds were fed a common starter to 14 D post-hatch. The experimental diets were provided subsequently until 25 D, which were a conventional corn-soy basal diet (CS) and a corn-soy diet displaced with 40% of corn (CN). These were fed as-is or supplemented with 40, 80, 120, or 160 kilo-Novo α-amylase units (KNU)/kg. Dietary treatments were distributed factorially as a 2 × 5 arrangement (diet type vs. amylase). Samples of feed, excreta, and ileal digesta were analyzed for determination of ileal digestible energy (IDE), ME, total tract retention, and digestibility of dry matter, protein, and fat. No interactions between diet type and amylase were observed. The CN diet had lower (P < 0.05) energy utilization and nutrient digestibility when compared to the CS diet. AMEn and IDE increased (P < 0.05) by 110 and 207 kcal/kg, respectively, when CS and CN diets were supplemented with 80 KNU/kg. The amylase added to the CS diet led to quadratic increases (P < 0.05) on growth performance, IDE, AME, and AMEn, as well as in dry matter, protein, and fat digestibility. Energy utilization and crude protein digestibility were linearly increased (P < 0.05) when amylase was added to the CN diet or the extrapolated 100% of corn. In conclusion, energy utilization, digestibility of crude protein, fat, and dry matter increased with amylase supplementation in corn-soy-based diets. When amylase was tested in a complete diet having 53.6% corn, 100 and 105 KNU/kg maximized AMEn and IDE, respectively; however, the maximum energy response in the CN diet or 100% of corn was not achieved until 160 KNU/kg, suggesting an association between amylase dose optimization and dietary starch concentration., (© 2019 Poultry Science Association Inc.)

In this study, methacrylated gellan-gum (GG-MA) heteropolysaccharide is proposed as a hydrogel for drug delivery and bone tissue engineering applications. Calcium-enriched beads obtained from the crosslinking of 1% (w/v) GG-MA solutions with 0.1 MCaCl 2 were investigated, considering their intrinsic capacity to promote self-mineralization by ion binding and deposition. Indeed, when immersed in a physiological environment, the Ca-enriched beads promoted the development of a bone-like apatite layer, as confirmed by EDS and XRD chemical analysis. Additionally, the mild production process is compatible with drugs incorporation and release. After encapsulation, Dextran with different molecular weights as well as Dexamethasone 21-phosphate were efficiently released to the surrounding environment. The engineered system was also evaluated considering its biocompatibility, by means of qualitative determination of total complement activation, macrophage proliferation, cytokine release and in vitro cell culture. These experiments showed that the developed hydrogels may not stimulate a disproportionate pro-inflammatory reaction once transplanted. At last, when implanted subcutaneously in CD1 male mice up to 8 weeks, the beads were completely calcified, and no inflammatory reaction was observed. Summing up, these results show that calcium-enriched GG-MA hydrogel beads hold great potential as news tools for bone tissue regeneration and local drug delivery applications. STATEMENT OF SIGNIFICANCE: This work describes a low-cost and straightforward strategy to prepare bioactive methacrylated gellan gum (GG-MA) hydrogels, which can be used as drug delivery systems. GG-MA is a highly anionic polymer, that can be crosslinked with divalent ions, as calcium. Taking advantage of this feature, it was possible to prepare Ca-enriched GG-MA hydrogel beads. These beads display a bioactive behavior, since they promote apatite deposition when placed in physiological conditions. Studies on the immune response suggest that the developed beads do not trigger severe immune responses. Importantly, the mild processing method render these beads compliant with drug delivery strategies, paving the way for the application of dual-functional materials on bone tissue engineering., (Copyright © 2019 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

One hundred and twenty Cobb 500 hens, 20 wk of age, were randomly allocated into individual cages with the objective of estimating their Zn requirements. The study was composed of 3 phases: adaptation to cages (basal diet), depletion (deficient diet containing 18.7 ± 0.47 ppm Zn) for 7 wk, and experimental phases. Hens were fed diets with graded increments of Zn sulfate heptahydrate (ZnSO4·7H2O), totaling 18.7 ± 0.47, 50.3 ± 10.6, 77.3.0 ± 11.0, 110.2 ± 12.8, 140 ± 12.2, and 170.6 ± 13.2 ppm analyzed Zn in feeds for 12 wk (experimental phase). Requirements of Zn were done using quadratic polynomial (QP), broken line quadratic (BLQ), and exponential asymptotic (EA) models. In general, the non-linear statistical models were the ones that best fit the results in this study. Requirements obtained for hen day egg production and settable egg production were 83.3, 78.6 ppm and 61.4, 65.4 ppm for period of 33 to 36 wk, and 63.3, 53.1 and 60.4, 46.1 ppm for period of 37 to 40 wk, and 62.8, 52.8, and 67.7, 62.1 ppm for period of 41 to 44 wk, respectively, using BLQ and EA models. Total eggs and total settable eggs produced per hen had Zn requirements estimated as 75.7, 64.7 ppm, and 56.5, 41.5 ppm, respectively, for BLQ and EA models, whereas for alkaline phosphatase and eggshell percentage were 161.8, 124.9 ppm and 126.1, 122.4 ppm, using QP and BLQ models. Maximum responses for Zn in yolk for periods of 37 to 40 and 41 to 44 wk were 71.0, 78.1 and 64.5, 59.6 ppm, respectively, using BLQ and EA models. Breaking strength had Zn requirements estimated at 68.0 and 96.7 ppm, whereas eggshell palisade layer and eggshell thickness were maximized with 67.9, 67.9 ppm, and 67.7, 64.4 ppm, respectively, for BLQ and EA models. The average of all Zn requirement estimates obtained by EA and BLQ models in the present study was 72.28 ppm or 11.1 mg/hen/d., (© 2018 Poultry Science Association Inc.)

Food chain information (FCI) is a mandatory component of meat inspection in the European Union. In Portugal there has been no assessment of FCI regarding risk-based meat inspection. The goal of the present study was to assess the quality and the usefulness of data provided by farmers. A total of 1,694 reports (989 for bovine animals, 575 for swine, and 130 for small ruminants) submitted to nine red meat slaughterhouses between September 2015 and January 2017 were analyzed. These reports covered a total of 79,889 animals. Overall, information was provided for the majority of the general identification items. To assess the quality of the information on health status, medication, disease occurrence, diagnostic tests, and results of previous ante- and postmortem exams, responses were classified as plausible, "nothing to declare," invalid, and nonresponse. Plausible information was provided by farmers on health status, medication, tests, and diseases on 82.7, 24.6, 5.4, and 0.1% of the FCI forms, respectively. More than 70% of the responses in all categories except health status were "nothing to declare." In pigs, when comparing ante- and postmortem inspection results with the corresponding FCI, no relevant contribution of the FCI was observed. Inaccurate or irrelevant information was provided for the majority of the detailed questions, impeding the formulation of appropriate risk-based meat inspection decisions. Farmers seem to have difficulty providing specific information. Better forms and better training are needed. Information on the prevalence of biological hazards relevant to public health currently is not available. Our results support the need to improve the systems used to obtain and integrate relevant information from the farm to the slaughterhouse.

One-hundred-twenty Cobb 500 hens, 20 wk of age, were randomly allocated into individual cages with the objective of estimating Cu requirements. After being fed a Cu deficient diet for 4 wk, hens were fed diets with graded increments of supplemental Cu (0.0; 3.5; 7.0; 10.5; 14; and 17.5 ppm) from Cu sulfate (CuSO4 5H2O), totaling 2.67; 5.82; 9.38; 12.92; 16.83; and 20.19 ppm analyzed Cu in feeds for 20 weeks. Estimations of Cu requirements were done using exponential asymptotic (EA), broken line quadratic (BLQ), and quadratic polynomial (QP) models. Obtained Cu requirements for hen d egg production and total settable eggs per hen were 6.2, 7.3, and 12.9 ppm and 8.1, 9.0, and 13.4 ppm, respectively, using EA, BLQ, and QP models. The QP model was the only one having a fit for total eggs per hen with 13.1 ppm Cu as a requirement. Hemoglobin, hematocrit, and serum Cu from hens had requirements estimated as 13.9, 11.3, and 18.5, ppm; 14.6, 13.0, and 19.0 ppm; and 16.2, 14.6, and 14.2 ppm, respectively, for EA, BLQ, and QP models. Hatching chick hemoglobin was not affected by dietary Cu, whereas requirements estimated for hatching chick hematocrit and body weight and length were 10.2, 12.3, and 13.3 ppm using EA, BLQ, and QP models; and 6.8 and 7.1 ppm, and 12.9 and 13.9 ppm Cu using EA and BLQ models, respectively. Maximum responses for egg weight, yolk Cu content, and eggshell membrane thickness were 14.9, 12.7, and 15.1 ppm; 15.0, 16.3, and 15.7 ppm; and 7.3, 7.8, and 14.0 ppm Cu, respectively, for EA, BLQ, and QP models. Yolk and albumen percentage were adjusted only with the QP model and had requirements estimated at 11.0 ppm and 11.3 ppm, respectively, whereas eggshell mammillary layer was maximized with 10.6, 10.1, and 14.4 ppm Cu using EA, BLQ, and QP models, respectively. The average of all Cu requirement estimates obtained in the present study was 12.5 ppm Cu.

Fish sarcoplasmic proteins (FSP) constitute around 25-30% of the total fish muscle protein. As the FSP are water soluble, FSP were isolated from fresh cod (Gadus morhua) by centrifugation. By SDS-PAGE, it was possible to determine the composition of FSP extracts (FSP-E). The FSP-E undergo denaturation at 44.12 ± 2.34° C, as characterized by differential scanning calorimetry thermograms (DSC). The secondary structure of FSP-E is mainly composed by α-helix structure, as determined by circular dichroism. The cytocompatibility of FSP-E, at concentrations ranging from 5 to 20 mg/mL, was investigated. Concentrations lower than 10 mg/mL have no cytotoxicity cultures of fibroblasts over 72 h. Further on, FSP membranes (FSP-M) were produced by spin coating to evaluate its properties. FSP-M shown having uniform surface as analyzed by Scanning Electron Microscopy (SEM). The relative amount of α-helix structures is higher when compared with the FSP-E. The FSP-M have higher temperature stability than the FSP-E, since they presented a denaturation temperature of 58.88 ± 3.36° C, according to the DSC analysis. FSP-M shown distinctive mechanical properties, with a stiffness of 16.57 ± 3.95 MPa and a yield strength of 23.85 ± 5.97 MPa. Human lung fibroblasts cell lines (MRC-5) were cultured in direct contact with FSP-M, demonstrating its cytocompatibility for 48 h. Based on these results, FSP can be considered a potential biomaterial recovered from nature, for wound dressing applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A study was conducted to evaluate growth performance, carcass and breast yields, and the occurrence and severity of white striping (WS) and wooden breast (WB) myopathies of broilers fed diets supplemented with increasing dietary levels of an organic source of selenium (Zn-L-SeMet). Broilers were fed 6 treatments with 12 replications of 26 birds in a 4-phase feeding program from 1 to 42 days. Corn-soy-based diets were supplemented with 0, 0.2, 0.4, 0.6, 0.8, and 1.0 ppm of Zn-L-SeMet. At 42 d, 6 birds were randomly selected from each pen (n = 72) and processed for carcass and breast yields. Breast fillets were scored for WS and WB at 42 days. Increasing Zn-L-SeMet led to quadratic responses (P < 0.05) for FCR from 1 to 7 d, BWG from 22 to 35 d, and for both responses from 8 to 21 d and 36 to 42 d, as well as in the overall period of 42 days. Carcass and breast yields presented a quadratic improvement (P < 0.01) with increasing Zn-L-SeMet supplementation and Se requirements were estimated at 0.85 and 0.86 ppm, respectively. In the overall period, estimates of Se requirements were 0.64 ppm for BWG and 0.67 ppm for FCR. White striping and WB scores presented quadratic increases (P < 0.01), and maximum scores were observed at 0.68 and 0.67 ppm, respectively. Broilers fed diets formulated without Se supplementation had a higher percentage of normal fillets compared to other Se supplementation levels (quadratic, P < 0.05). In conclusion, increasing Se supplementation to reach maximum growth performance led to higher degrees of severity of WS and WB. Selenium requirements determined in the present study were significantly higher than the present commercial recommendations.

Sporotrichosis is caused by species of fungi within the Sporothrix schenckii complex that infect man and animals. In Rio de Janeiro, Brazil, an epidemic has been observed since 1998, with most of the cases being related to transmission from infected cats. Although the definitive diagnosis of feline sporotrichosis is made by fungal culture, cytopathological and histopathological examinations are used routinely, because the long culture period may delay treatment onset. However, alternative methods are desirable in cases of low fungal burden. Immunohistochemistry (IHC) has been described as a sensitive method for diagnosing human and canine sporotrichosis, but there are no reports of its application to cats. The aim of this study was to analyse the sensitivity of cytopathological examination (Quick Panoptic method), histopathology (Grocott silver stain) and anti-Sporothrix IHC by blinded comparisons, using fungal culture as the reference standard. Samples were collected from 184 cats with sporotrichosis that exhibited skin ulcers. The sensitivities of Grocott silver stain, cytopathological examination and IHC were 91.3%, 87.0% and 88.6%, respectively. Grocott silver stain showed the best performance. IHC showed high sensitivity, as did cytopathological examination and these may be considered as alternative methodologies. When the three methods were combined, the diagnosis was established in 180 (97.8%) out of 184 cases. Taken together, these findings indicate the need to implement these methods as routine tools for the early diagnosis of sporotrichosis in cats, notably when fungal culture is not available., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

As the first limiting amino acid in corn-soy broiler diets, methionine (Met) is supplemented using commercial synthetic sources as demanded to obtain economic feed formulations. The Met analogue DL-2-hydroxy-4-(methylthio)-butanoic acid (HMTBA) is largely utilized with that objective. This study intended to obtain responses of broilers fed with increasing levels of HMTBA, from 28 to 42 d, such that economic returns can be calculated. A total of 2,106 Cobb × Cobb 500 one-day-old male broilers was randomly placed in 81 floor pens (2.7 m2 each). Birds were fed conventional starter (zero to 14 d) and grower (14 to 28 d) diets. Starting at 28 d of age, pens of 26 birds were randomly allocated into 9 feed treatments with 9 replications having increasing supplementations with HMTBA (0.00, 0.07, 0.14, 0.21, 0.28, 0.35, 0.42, 0.49 and 0.56%). These were prepared by mixing different proportions of corn-soy dilution and summit diets, which had the same formulated concentration of nutrients and energy [19.7% CP, 0.90% Ca, 0.45% Av. P, 0.95% digestible Lys, and 3,150 kcal/kg AMEn], with the exception of HMTBA [0.56% in the summit but not supplemented in the corn-soy dilution diet (0.52% digestible TSAA)]]. Growth performance was evaluated until 42 d when carcass yield and commercial cuts were evaluated using 6 birds randomly taken from each pen. Body weight gain (BWG), feed conversion ratio (FCR), proportion of breast fillets, and abdominal fat were adjusted using linear broken-line, exponential asymptotic and quadratic polynomial regression models (P < 0.05). Estimations of maximum responses for supplemented HMTBA by the linear broken-line model were 0.17% for BWG, 0.14% for FCR, and 0.29% for breast fillets. Using exponential and quadratic regressions, optimized HMTBA supplementations were obtained at 0.34 and 0.35% for BWG, 0.20 and 0.33% for FCR, and 0.31 and 0.36% for breast fillets, respectively. Supplemental levels of HMTBA that optimize growth performance and breast meat in male broilers from 28 to 42 d, using different regression models, varied from 0.14 to 0.36%.

The pro-apoptotic Bax protein is the main effector of mitochondrial permeabilization during apoptosis. Bax is controlled at several levels, including post-translational modifications such as phosphorylation and S-palmitoylation. However, little is known about the contribution of other protein modifications to Bax activity. Here, we used heterologous expression of human Bax in yeast to study the involvement of N-terminal acetylation by yNaa20p (yNatB) on Bax function. We found that human Bax is N-terminal (Nt-)acetylated by yNaa20p and that Nt-acetylation of Bax is essential to maintain Bax in an inactive conformation in the cytosol of yeast and Mouse Embryonic Fibroblast (MEF) cells. Bax accumulates in the mitochondria of yeast naa20Δ and Naa25 -/- MEF cells, but does not promote cytochrome c release, suggesting that an additional step is required for full activation of Bax. Altogether, our results show that Bax N-terminal acetylation by NatB is involved in its mitochondrial targeting., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A study was conducted to determine the AMEn contents of fat by-products from the soybean oil industry for broiler chickens. A total of 390 slow-feathering Cobb × Cobb 500 male broilers were randomly distributed into 13 treatments having 6 replicates of 5 birds each. Birds were fed a common starter diet from placement to 21 d. Experimental corn-soy diets were composed of four fat sources, added at 3 increasing levels each, and were fed from 21 to 28 d. Fat sources utilized were acidulated soybean soapstock (ASS), glycerol (GLY), lecithin (LEC), and a mixture (MIX) containing 85% ASS, 10% GLY and 5% LEC. A 4 × 3 + 1 factorial arrangement was used with 4 by-products (ASS, GLY, LEC, or MIX), 3 inclusion levels and 1 basal diet. Each of the four fat by-product sources was included in the diets as follow: 2% of by-products (98% basal + 2% by-product), 4% (96% basal + 4% by-product), or 6% (94% basal + 6% by-product). Birds were submitted to 94, 96, 98, and 100% of ad libitum feed intake; therefore, the differences in AMEn consumption were only due to the added by-product. Total excreta were collected twice daily for 72 h to determine apparent metabolizable energy contents starting at 25 d. The AMEn intake was regressed against feed intake and the slope was used to estimate AMEn values for each fat source. Linear regression equations (P < 0.05) estimated for each by-product were as follow: 7,153X - 451.9 for ASS; 3,916X - 68.2 for GLY; 7,051X - 448.3 for LEC, and 8,515X - 622.3 for MIX. Values of AMEn were 7,153, 3,916, 7,051, and 8,515 kcal/kg DM for ASS, GLY, LEC, and MIX, respectively. The present study generated AMEn for fat by-products data that can be used in poultry feed formulation. It also provides indications that, by adding the 3 by-products in the proportions present in the MIX, considerable economic advantage can be attained., (© 2017 Poultry Science Association Inc.)

Dorsal cranial myopathy (DCM), which affects the anterior latissimus dorsi (ALD) muscles of commercial broilers, is of unknown etiology, and it represents up to 6% of the partial condemnations in Brazilian slaughterhouses. This study was performed to achieve histomorphometric characterizations of the ALD muscles from male Cobb 500 broilers slaughtered at either 35 d or 42 d and to evaluate the effects of DCM on the enzymatic markers aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), and lactate dehydrogenase (LDH) and on uric acid and creatinine metabolites. Blood samples (1.5 to 3 mL) and ALD muscle fragments were collected from each carcass, all of which were processed in a commercial inline processing system. For each age, twelve macroscopically normal animals and twelve animals found to exhibit DCM were randomly selected for histomorphometric evaluation and analysis of serologic profiles. Microscopic evaluations demonstrated that the muscle fibers of those with DCM exhibited a strong presence of multifocal regenerative myodegeneration as well as a substitution of muscle tissue with connective tissue (P < 0.001) through fibrosis, thus characterizing the chronicity and hardness of the affected muscle. It is suggested that DCM is a localized muscle lesion because the detected serum levels of CK (P < 0.001), AST (P < 0.001), ALT (P = 0.01), and LDH (P < 0.001) enzymes were strongly associated with the group affected by DCM. Additional studies are needed to gain an understanding of this myopathy because it is an emerging problem in the poultry industry. In addition, it is related to DCM lesions in fast-growing broilers with the greatest slaughter weights., (© 2017 Poultry Science Association Inc.)

Background: We investigated whether the treatment with enalapril maleate, combined with aerobic physical training, promotes positive effects on the autonomic balance, the morphology and the cardiac function in female rats submitted to early ovarian failure., Methods: Thirty-five female Wistar rats, ovariectomized at 10weeks of age, were assigned into Ovariectomized rats (OVX) and Ovariectomized rats treated with enalapril maleate (OVX-EM, 10mg -1 ·kg -1 ·d -1 ) Each group was subdivided into sedentary and trained (aerobic swimming training for 10weeks). All animals were submitted to a) double pharmacological autonomic blockade, b) study of morphology and cardiac function by echocardiography, and c) analysis of cardiac fibrosis., Results: The OVX-EM sedentary group showed a significant increase in cardiac fibrosis, relative heart weight, interventricular septum thickness and increased sympathetic participation and reduced participation of the vagal tone in the determination of the basal heart rate when compared to the OVX sedentary group. Physical training reduced cardiac fibrosis in both groups, however, with less intensity in the OVX-EM group. It also increased the absolute and relative heart weight and the end-systolic volume. Finally, the OVX-EM trained group showed higher values for left ventricular end-systolic volume and lower values for ejection fraction and shortening fraction than the sedentary OVX-EM group., Conclusion: Enalapril maleate exacerbated cardiac fibrosis and increased sympathetic participation in the basal heart rate determination, without significantly affecting the cardiac function. Aerobic physical training did not change the cardiac autonomic control, but reduced cardiac fibrosis and had little effect on the cardiac function., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Two experiments were conducted to evaluate the prevalence and severity of white striping (WS) and wooden breast (WB) in breast fillets from broilers fed diets with increasing digestible Lysine (dLys) from 12 to 28 d (Exp. 1) and from 28 to 42 d (Exp. 2). Trials were sequentially conducted using one-d-old male, slow-feathering Cobb 500 × Cobb broilers, both with 6 treatments and 8 replicates. Increasing dLys levels were equally spaced from 0.77 to 1.17% in Exp. 1 and from 0.68 to 1.07% in Exp. 2. The lowest dLys diet was not supplemented with L-Lysine (L-Lys) in either one of the studies and all other essential amino acid (AA) met or exceeded current commercial recommendations such that their dietary concentrations did not limit broiler growth. Four birds per pen were randomly selected from each replication and processed at 35 and 42 d in Exp. 1 and Exp. 2, respectively. Deboned breast fillets (Pectoralis major) were submitted to a 3 subject panel evaluation to detect the presence of WS and WB, as well as to provide scores of WS (0-normal, 1-moderate, 2-severe) and WB (0-normal, 1-moderate light, 2-moderate, 3-severe). Increasing the level of dLys had a positive effect on BW, carcass, and breast weight, as well as breast yield. White striping and WB prevalences were 32.3 and 85.9% in Exp. 1 and 87.1 and 89.2% in Exp. 2. Birds fed diets not supplemented with L-Lys had the lowest average WS and WB scores (0.22 and 0.78 in Exp. 1 and 0.61 and 0.68 in Exp. 2). White striping and WB presented linear responses to performance variables in Exp. 1, whereas quadratic responses were observed for all variables in Exp. 2. In conclusion, increasing the level of dLys improved growth performance and carcass traits as well as induced the occurrence and severity of WS and WB lesions., (© 2016 Poultry Science Association Inc.)

A study was conducted to evaluate the effects of increased levels of a β-xylanase on energy and nutrient utilization of broiler chickens fed corn-soy diets. A total of 480 slow feathering Cobb × Cobb 500 male broilers were randomly distributed to 10 treatments having 8 replicates of 6 birds each. Birds were fed a common starter diet to d 14 post hatch (3,050 kcal/kg AMEn, 21.7% CP, 1.05% Ca, and 0.53% nPP). The experimental diets were provided afterwards until 25 d. Two experimental diets, a conventional corn/soy-based basal diet (CS) and the basal diet in which 40% of the diet was displaced by corn (CN), were fed as-is or supplemented with 50, 100, 150, or 200 fungal β-xylanase units (FXU)/kg. Dietary treatments were distributed factorially as a 2 × 5 arrangement. Samples of feed, excreta, and ileal digesta were analyzed for determination of ileal digestible energy (IDE), metabolizable energy, and total tract retention of protein and lipid. No interactions between diet and xylanase were observed. The CS diets had higher (P < 0.05) energy utilization and nutrient digestibility when compared to the CN diets. AMEn and IDE were improved (P < 0.05) by 192 and 145 kcal/kg, respectively, when diets were supplemented with 100 FXU/kg xylanase. The xylanase added to the CN diet led to quadratic increases (P < 0.05) in IDE (Y = - 0.014x(2) + 2.570x + 3,155; r(2) = 0.60) and in AMEn (Y = - 0.016x(2) + 3.982x + 3,155; r(2) = 0.68). Crude protein digestibility and AMEn were linearly increased (P < 0.05) when xylanase was added to the CN diet. In conclusion, energy utilization and digestibility of crude protein and dry matter increased with xylanase supplementation in corn/soy-based diets. When xylanase was tested in the CS diet, 92 and 124 FXU/kg maximized the energy release effect; however, the maximum energy response in the CN diet or corn was not achieved until 200 FXU/kg., (© 2016 Poultry Science Association Inc.)

A study was conducted to determine the ileal digestible energy (IDE), ME, and MEn contents of bakery meal using the regression method and to evaluate whether the energy values are age-dependent in broiler chickens from zero to 21 d post hatching. Seven hundred and eighty male Ross 708 chicks were fed 3 experimental diets in which bakery meal was incorporated into a corn-soybean meal-based reference diet at zero, 100, or 200 g/kg by replacing the energy-yielding ingredients. A 3 × 3 factorial arrangement of 3 ages (1, 2, or 3 wk) and 3 dietary bakery meal levels were used. Birds were fed the same experimental diets in these 3 evaluated ages. Birds were grouped by weight into 10 replicates per treatment in a randomized complete block design. Apparent ileal digestibility and total tract retention of DM, N, and energy were calculated. Expression of mucin (MUC2), sodium-dependent phosphate transporter (NaPi-IIb), solute carrier family 7 (cationic amino acid transporter, Y(+) system, SLC7A2), glucose (GLUT2), and sodium-glucose linked transporter (SGLT1) genes were measured at each age in the jejunum by real-time PCR. Addition of bakery meal to the reference diet resulted in a linear decrease in retention of DM, N, and energy, and a quadratic reduction (P < 0.05) in N retention and ME. There was a linear increase in DM, N, and energy as birds' ages increased from 1 to 3 wk. Dietary bakery meal did not affect jejunal gene expression. Expression of genes encoding MUC2, NaPi-IIb, and SLC7A2 linearly increased (P < 0.05) with age. Regression-derived MEn of bakery meal linearly increased (P < 0.05) as the age of birds increased, with values of 2,710, 2,820, and 2,923 kcal/kg DM for 1, 2, and 3 wk, respectively. Based on these results, utilization of energy and nitrogen in the basal diet decreased when bakery meal was included and increased with age of broiler chickens., (© 2016 Poultry Science Association Inc.)

Aims: We systematically reviewed the role of low-level laser therapy (LLLT) in cardiac remodeling after myocardial infarction., Main Methods: Literatures were systematically searched in several electronic databases. We included only studies with a well-standardized coronary occlusion model in vivo LLLT application., Key Findings: After screening, 14 studies were eligible for review. The study heterogeneity was described in terms of rationality, gender, irradiation parameters, treatment numbers and moment of LLLT application. Three studies showed a null role of LLLT on infarct size, and only one study found positive LLLT effects on the cardiac performance. The cardioprotective role of LLLT was mediated by anti-inflammatory, pro-angiogenic and anti-oxidant actions., Significance: The reduction in infarct size is a major finding. The LLLT cardioprotection may be mediated by several molecular and cellular mechanisms. Although these results are exciting, there are many limitations that must be resolved before LLLT clinical trials., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The role of lysine 369 of human topoisomerase IB in recognizing, clamping and processing its DNA substrate was experimentally investigated. Lys(369) is located in one of the two lips that interact to each other allowing the protein to embrace and firmly bind the DNA substrate. The lysine was mutated to a glutamate residue and the catalytic activity of the mutant enzyme was assayed. The mutant shows a distributive behavior, has a reduced binding to the substrate and a lower cleavage extent when compared to the wild type enzyme. The mutant displays reduced sensitivity to CPT both "in vitro" and in an "in vivo" yeast model, likely because of the low amount of cleaved DNA, however it displays cleavage and religation rates comparable to the wild type. These results demonstrate that the mutation causes a destabilization of the lips clamping around the DNA, impairing the protein-DNA interaction, emphasizing the importance of the ionic pair in tuning the stability of the protein-DNA complex., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A study was conducted to evaluate the effects of dietary α-amylase and β-xylanase supplementation of corn-soy diets, formulated with or without supplemental phytase, on growth performance, energy utilization, and starch digestibility in broiler chickens. A total of 336 slow-feathering, Cobb × Cobb 500 male broilers were randomly distributed to 6 treatments having 8 replicates of 7 birds each. Birds were fed a common starter diet to d 14 post-hatch (3,050 kcal/kg AMEn, 21.7% CP, 1.05% Ca, and 0.53% nPP). The experimental diets were provided afterwards until d 25. A 2 × 3 factorial arrangement of 2 control diets (basal = corn-soy diet without added phytase or PHY = corn-soy diet formulated with 1,000 phytase units/kg) and 3 carbohydrase supplementations (0, 80 kilo-Novo α-amylase units/kg, or 80 kilo-Novo α-amylase units/kg + 100 fungal β-xylanase units/kg) was used from d 14 to 25. Excreta were collected from 21 to 24 d and all birds were euthanized at 25 d for jejunum and ileum content collection. Samples of feed, excreta, and jejunal and ileal digesta were analyzed for determination of total tract retention and ileal apparent digestibility. No interactions between diet and carbohydrase were observed. Broilers fed diets formulated with phytase or supplemented with amylase + xylanase had higher BW gain (BWG) and lower FCR (P < 0.05) when compared with birds fed diets without carbohydrases. Relative to the basal diet, AMEn was increased (P < 0.01) by 70 kcal/kg and 99 kcal/kg when birds were fed the diet supplemented with amylase and amylase + xylanase, respectively. Starch digestibility in the jejunum and ileum was increased (P < 0.05) by 3.5% and 2.4%, respectively, when birds were fed the diet supplemented with amylase + xylanase. Results from this experiment show that corn-soy diets having phytase and supplemented with amylase and xylanase led to increased growth performance, AMEn, and starch digestibility in broilers. Furthermore, the efficacy of exogenous amylase and xylanase was independent of the presence of microbial phytase., (© 2015 Poultry Science Association Inc.)

Topoisomerases IB are anticancer and antimicrobial targets whose inhibition by several natural and non-natural compounds has been documented. The inhibition effect by berberine and some 13-(di)phenylalkyl berberine derivatives has been tested towards human topoisomerase IB. Derivatives belonging to the 13-diphenylalkyl series display an efficient inhibition of the DNA relaxation and cleavage step, that increases upon pre-incubation with the enzyme. The religation step of the enzyme catalytic cycle is not affected by compounds and only slightly upon pre-incubation. The binding of the protein to the DNA substrate occurs also in the presence of the compounds, as monitored by a DNA shift assay, indicating that the compounds are not able to inhibit the formation of the enzyme-DNA complex but that they act as catalytic inhibitors., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Since it started as an organized economic activity, poultry production has been undergoing an evolution toward the optimization of its feed formulation features. Notably, advances in the knowledge of birds' nutrient requirements have allowed recommendations that are increasingly closer to the needs of the birds. Over time, availability of nutrients and energy has been incorporated into those recommendations, especially to compensate for the variability in the digestibility of nutrients originating from variable feed ingredient sources. Instead of using the total energy and nutrient content, current tables of nutrient recommendations provide an estimate of the digestible fractions of the nutrients in ingredients. For instance, nonphytate P is preferred instead of total P to account for the unavailable phytate P, and digestible amino acids to account for the differences in digestibility of amino acids in different ingredients, whereas energy is usually expressed as a proportion that has been digested and metabolized (AME). With the increasing interest in the use of exogenous enzymes in poultry feeds, special attention is directed to the feed substrates such that an added enzyme can match it, forming an enzyme-substrate complex that will be followed by a chemical reaction within the gastrointestinal tract. As a consequence of a degradation reaction, nutrients released can be absorbed and metabolized. In general, nutritional data banks used in linear feed formulation software have limited data on the proportions of fractions of ingredients that are indigestible. Therefore, estimations of the presence of many substrates in the feeds, and therefore the benefits of adding exogenous enzymes, are frequently limited because of the scarcity of adequate information. The objective of this review paper is to provide insights into the use of expanded nutrient databanks to include all the molecules considered potentially indigestible for poultry such that the inclusion of exogenous enzymes allows the estimation of the values of the product originated by their hydrolysis., (© 2014 Poultry Science Association Inc.)

Doping calcium phosphates with trace elements that exist in bone tissues is beneficial in terms of cell-material interactions and in vivo performance of the bone grafts made thereof. Manganese (Mn) is an essential element for normal growth and metabolism of bone tissues, but studies reporting the effects of Mn-doping calcium phosphates are scarce. The present study investigated the influence of Mn-doping on the structure, morphology and biological properties of β-tricalcium phosphate [β-Ca3(PO4)2] (β-TCP). Mn-doped (MnTCP) powders, with Mn contents varying from 0 to 10 mol%, were obtained through an aqueous precipitation method followed by heat treatment at 800 °C. The successful incorporation of Mn into β-TCP structure was proved through quantitative X-ray diffraction (XRD) phase analysis coupled with structural Rietveld refinement. Increasing Mn concentrations led to decreasing trends of a- and c-axis lattice parameters, and Mn-doping also significantly affected the morphology of β-TCP powders. In vitro proliferation and differentiation assays of MC3T3-E1 osteoblastic-like cells, grown in the presence of the powders, revealed that the biological benefits of Mn-doped β-TCP are limited to lower Mn incorporation levels and potentially related to their surface microstructure. The Mn1-βTCP composition revealed the best set of bioactivity properties, potentially a good candidate for future applications of β-TCP materials in osteoregeneration., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objectives: To assess in situ the effect of different restorative techniques used with fluoride-releasing materials on enamel and dentine demineralization in the presence of a cariogenic challenge., Methods: Thirty human molars were prepared for 60 Class V cavities, with enamel and dentine margins. The teeth were divided into four groups (n=15): L1 - open sandwich technique with a conventional glass ionomer cement (GIC), L2 - open sandwich technique with a resin-modified GIC, A1 - total-etch bonding with a fluoride-containing adhesive, A2 - total-etch bonding with a nonfluoride-containing adhesive. All the cavities were restored with a nanofilled composite. Fifteen volunteers used appliances containing one specimen from each group. The cariogenic challenge was carried out with a 20% sucrose solution 8×/day for 7 days. The specimens were sectioned for microhardness test and EDS analysis at different depths below the enamel and dentine margins (25, 50 and 100μm) and distances from the tooth-restoration interfaces (25 and 75μm). The data were analyzed by ANOVA and Games-Howell test (α=5%)., Results: Both GICs produced higher hardness in enamel at all depth-and-distance combinations, but only L1 produced higher hardness in dentine (p<0.05). L2 and A1 exhibited similar dentine hardness at 25μm distance for all depths (p>0.05). L1 demonstrated significantly higher amounts of calcium in dentine than the other groups, but had similar amounts in enamel to L2 and A1., Conclusions: The open sandwich technique using conventional GIC proved more effective in reducing enamel and dentine demineralization at depths of up to 100μm under a cariogenic challenge., Clinical Significance: Conventional GICs should be considered the materials of choice for lining of cavities not having all margins in enamel, particularly using the open sandwich technique., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Snake venom metalloproteinases (SVMPs) participate in a number of important biological, physiological and pathophysiological processes and are primarily responsible for the local tissue damage characteristic of viperid snake envenomations. The use of medicinal plant extracts as antidotes against animal venoms is an old practice, especially against snake envenomations. Such plants are sources of many pharmacologically active compounds and have been shown to antagonize the effects of some venoms and toxins. The present study explores the activity of triacontyl p-coumarate (PCT), an active compound isolated from root bark of Bombacopsis glabra vegetal extract (Bg), against harmful effects of Bothropoides pauloensis snake venom and isolated toxins (SVMPs or phospholipase A(2)). Before inhibition assays, Bg or PCT was incubated with venom or toxins at ratios of 1:1 and 1:5 (w/w; venom or isolated toxins/PCT) for 30 min at 37°C. Treatment conditions were also assayed to simulate snakebite with PCT inoculated at either the same venom or toxin site. PCT neutralized fibrinogenolytic activity and plasmatic fibrinogen depletion induced by B. pauloensis venom or isolated toxin. PCT also efficiently inhibited the hemorrhagic (3MDH - minimum hemorrhagic dose injected i.d into mice) and myotoxic activities induced by Jararhagin, a metalloproteinase from B. jararaca at 1:5 ratio (toxin: inhibitor, w/w) when it was previously incubated with PCT and injected into mice or when PCT was administered after toxin injection. Docking simulations using data on a metalloproteinase (Neuwiedase) structure suggest that the binding between the protein and the inhibitor occurs mainly in the active site region causing blockade of the enzymatic reaction by displacement of catalytic water. Steric hindrance may also play a role in the mechanism since the PCT hydrophobic tail was found to interact with the loop associated with substrate anchorage. Thus, PCT may provide a alternative to complement ophidian envenomation treatments., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Topoisomerases IB are anticancer and antimicrobial targets whose inhibition by several natural and synthetic compounds has been documented over the last three decades. Here we show that kakuol, a natural compound isolated from the rhizomes of Asarum sieboldii, and a derivative analogue are able to inhibit the DNA relaxation mediated by the human enzyme. The analogue is the most efficient one and the inhibitory effect is enhanced upon pre-incubation with the enzyme. Analysis of the different steps of the catalytic cycle indicates that the inhibition occurs at the cleavage level and does not prevent DNA binding. Molecular docking shows that the compound preferentially binds near the active site at the bottom of the catalytic residue Tyr723, providing an atomistic explanation for its inhibitory activity., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Sources of Zn, Mn, and Cu (IZMC) as sulfates or as amino acid complexes (OZMC) were used to supplement Cobb 500 breeder hen diets. Experimental treatments consisted of diets supplemented with 1) 100, 100, and 10 mg/kg of Zn, Mn, and Cu, respectively, from IZMC (control); 2) 60, 60, and 3 mg/kg of Zn, Mn, and Cu, respectively, from IZMC plus 40, 40, and 7 mg/kg of Zn, Mn, and Cu, respectively, from OZMC (ISO); and 3) a diet with 100, 100, and 10 mg/kg of Zn, Mn, and Cu, respectively, from IZMC as in control plus 40, 40, and 7 mg/kg of supplemental Zn, Mn, and Cu from OZMC (on top). Ten replications of 20 females and 2 males were used per treatment. Eggs from breeders at 30, 40, 50 and 60 wk of age were incubated, and 5 embryos per replicate were collected at 10 (E10), 14 (E14), and 18 (E18) d of incubation. Midshaft width and calcification were measured for left tibia and femur stained with Alcian Blue and Alizarin Red S. At hatch, the left tibia of 5 chicks per replicate was sampled for histological evaluation of the diaphysis and distal epiphysis. Feeding the ISO treatment compared with the control diet increased the Zn (P < 0.05) but not Mn and Cu content of the yolk and albumen blend. At E14, the ISO and on-top treatments had a trend to increase tibia calcification at the rates of 1.6 and 1%, respectively (P < 0.1). The E18 ISO and on-top treatments had 2% thicker tibia compared with the control, regardless of hen age (P < 0.05). Also, at E18, calcification of tibia and femur was higher from hens fed the on-top treatment (P < 0.05). The chicks from the ISO and on-top groups had increased tibia moment of inertia (P < 0.01) at day of hatch. Broiler breeder hens consuming OZMC associated with IZMC produced embryos and hatching chicks with improvements in selected bone mineralization parameters.

Introduction: The purpose of this study was to evaluate the reliability of measurements made on 3-dimensional digital models obtained with a surface laser scanner (D-250; 3Shape, Copenhagen, Denmark)., Methods: Twenty orthodontic dental casts of permanent dentitions were selected. Three-dimensional images were obtained on this scanner and analyzed by using the Geomagic Studio 5 software (Raindrop Geomagic, Inc, Morrisville, NC). Measurements were made with a digital caliper directly on the dental casts and also digitally on the digital models. Fifteen anatomic dental points were identified, and a total of 11 linear measurements were taken from each cast, including arch length and width. Dependent t tests were used to evaluate intraexaminer reproducibility and measurement accuracy on the digital models., Results: No statistically significant differences were found between the measurements made directly on the dental casts and on the digital models., Conclusions: Linear measurements on digital models are accurate and reproducible. Digital models obtained with the surface laser scanner are reliable for measurements of arch width and length., (Copyright © 2012 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.)

Background: The authors conducted a study to evaluate the stain removal ability of tooth bleaching and simulated toothbrushing after coffee and cigarette smoke staining and to determine the enamel susceptibility to restaining., Methods: The authors used a colorimeter to determine the baseline color of 40 bovine labial enamel surfaces according to the Commission Internationale de l'Eclairage L*a*b* coordinates. They immersed one-half of the specimens in coffee and exposed one-half to cigarette smoke in a smoking machine. They took color measurements again and determined the color change from baseline (ΔE1) for each group. The authors divided each group into two subgroups and subjected the specimens to at-home bleaching (one hour per day for 21 days) or simulated toothbrushing (120 cycles per day for 21 days), followed by another color measurement (ΔE2). The authors repeated both staining procedures (that is, cigarette smoke and coffee) and followed them with a third color measurement (ΔE3). They analyzed the data by using a two-way analysis of variance and the Tukey test (α = 5 percent)., Results: Both staining procedures resulted in similar values for ΔE1. The specimens stained with coffee and cigarette smoke exhibited a significant reduction in color change after bleaching (P < .05). However, toothbrushing resulted in a significantly reduced color change only for cigarette smoke-stained specimens (P < .001). The discoloration in coffee-stained specimens increased after restaining, irrespective of the stain removal method (P < .05)., Conclusions: The study results show that at-home bleaching removed both coffee and cigarette smoke staining. The restaining potential was greater for specimens stained with coffee than for those stained with cigarette smoke, regardless of the removal method used., Clinical Implications: Six percent hydrogen peroxide at-home bleaching was effective in removing stains caused by coffee or cigarette smoke. However, continued frequent consumption of coffee can increase the staining susceptibility of enamel.

Objectives: Colour changes of the luting material can become clinically visible affecting the aesthetic appearance of thin ceramic laminates. The aim of this in vitro study was to evaluate the colour stability and opacity of light- and dual-cured resin cements and flowable composites after accelerated ageing., Methods: The luting agents were bonded (0.2 mm thick) to ceramic disks (0.75 mm thick) built with the pressed-ceramic IPS Aesthetic Empress (n=7). Colour measurements were determined using a FTIR spectrophotometer before and after accelerated ageing in a weathering machine with a total energy of 150 kJ. Changes in colour (ΔE) and opacity (ΔO) were obtained using the CIE L*a*b* system. The results were submitted to one-way ANOVA, Tukey HSD test and Student's t test (α=5%)., Results: All the materials showed significant changes in colour and opacity. The ΔE of the materials ranged from 0.41 to 2.40. The highest colour changes were attributed to RelyX ARC and AllCem, whilst lower changes were found in Variolink Veneer, Tetric Flow and Filtek Z350 Flow. The opacity of the materials ranged from -0.01 to 1.16 and its variation was not significant only for Opallis Flow and RelyX ARC., Conclusions: The accelerated ageing led to colour changes in all the evaluated materials, although they were considered clinically acceptable (ΔE<3). Amongst the dual-cured resin cements, Variolink II demonstrated the highest colour stability. All the flowable composites showed proper colour stability for the luting of ceramic veneers. After ageing, an increase in opacity was observed for most of the materials., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

A study was conducted with an exogenous monocomponent protease added to corn-soybean meal diets fed to straight-run Ross 708 broilers from 7 to 22 d of age. Broilers were randomly placed into 42 battery pens (5 birds/pen) and allocated to 6 treatments with 7 replicates. A positive control diet (PC; 22.5% CP) and a low protein basal diet (20.5% CP) were formulated. Low protein diets (LP) comprised 98.67% of low protein basal diet and 1.33% Celite (indigestible marker and filler; Celite Corp., Lompoc, CA). Protease [75,000 PROT units/g; 1 PROT unit is defined as the amount of enzyme that releases 1 µmol of p-nitroaniline from 1 µM of substrate (Suc-Ala-Ala-Pro-Phe-p-nitroaniline per minute at pH 9.0 and 37°C] was added at the expense of Celite (0 mg/kg, LP0; 100 mg/kg, LP100; 200 mg/kg, LP200; 400 mg/kg, LP400; and 800 mg/kg, LP800) to create the LP diets (20.25% CP). At 22 d of age, ileal contents were collected from all birds for apparent CP and amino acid (AA) digestibility determinations. Broilers fed the PC diet were 7.5% heavier (P < 0.05) compared with those fed the LP0 diet. Birds fed the LP diets containing protease regardless of concentration grew as well as the birds fed the PC diet. Feed conversion was impaired (P < 0.05) in birds fed the LP0 and the LP100 diets compared with those fed the PC diet, but no difference was found between birds fed the PC diet and those fed diets containing more protease (LP200, LP400, and LP800). Digestibility of CP was increased (P < 0.05) in broilers fed the LP-supplemented diets compared with those fed either the PC or LP0 diets, but it was similar between those fed LP diets with any protease concentration. Digestibility of AA was not different between the PC and LP0 diets. The protease used in this study restored live performance and digestibility of CP (6.1%). When benefits in AA digestibility occurred, they were similar at all protease inclusions and averaged as follows: Arg, 3.5%; Ile, 3.2%; Lys, 5.4%; Thr, 7.8%; Asp, 6.5%; His, 3.3%; Cys, 4.6%; and Ser, 5.5%. Methionine was increased only at 400 and 800 mg/kg (6.5%) and Val was increased only at 200 and 800 mg/kg (5%).

Background: The authors conducted a study to quantify the amount of hydrogen peroxide remaining after in-office bleaching and to investigate the effectiveness of 35 percent sodium ascorbate (SA) on removal of the bleaching agent from dentin., Methods: The authors sectioned 70 extracted human third molars to form 4×4×2-millimeter blocks of dentin and then categorized them into eight groups as follows: group 1, no bleaching; group 2, bleaching (35 percent hydrogen peroxide) and daily measurement; group 3, bleaching plus one application of 35 percent SA for 60 minutes; group 4, bleaching plus one application of SA for 10 minutes; group 5, bleaching plus two applications of SA for 10 minutes each; group 6, bleaching plus two applications of SA for five minutes each; group 7, bleaching plus three applications of SA for one minute each; and group 8, bleaching plus two applications of SA for one minute each. They quantified the amount of residual hydrogen peroxide by using a colorimetric assay with horseradish peroxidase and o-phenylenediamine dihydrochloride as the enzyme and substrate, respectively., Results: The authors analyzed the data by using the Kruskal-Wallis test (α = .05). The results showed that no hydrogen peroxide remained after 120 hours in group 2 and at their respective measurement points in groups 5, 6, 7 and 8, whereas hydrogen peroxide had been removed only partially in groups 3 and 4., Conclusion: Two applications of 35 percent SA for one minute each produced the same result--complete removal of the bleaching agent--as that observed five days after tooth bleaching with 35 percent hydrogen peroxide without any application of SA., Clinical Implications: In an appropriate concentration, SA facilitates rapid elimination of residual peroxide up to two minutes after bleaching with 35 percent hydrogen peroxide. This finding suggests that clinicians can perform bonding procedures shortly after the bleaching procedure.

Background: To evaluate the influence of radiotherapy on the selenium serum levels of breast cancer patients., Patients and Methods: This prospective study includes 209 breast cancer patients treated by external beam radiotherapy from December 2007 until August 2008. Plasma selenium concentrations were determined before and at the end of the radiotherapeutic treatment. Age, clinical stage, prior chemotherapy, body mass index (BMI) and personal habits (smoking and alcoholism) were recorded for each patient., Results: The mean age was 61 years; the mean BMI was 28.7. One hundred and seventy-four patients (83.3%) were nonsmokers. One hundred and eighty-nine patients (90.4%) showed no drinking habits and 110 (52.6%) have no prior chemotherapy. Sixty patients (28.7%) were in clinical stage I, 141 (67.5%) in clinical stage II and 8 (3.8%) in clinical stage III. At the beginning of radiotherapy, the mean selenium value for all patients was 86.4 μg/l and after radiation this value dropped to 47.8 μg/l. Multivariate analysis showed statistically significant difference in the plasma selenium concentration before and after radiotherapy for age (P > 0.001), BMI (P > 0.001), smoking (P > 0.001), alcoholism (P > 0.001), chemotherapy (P > 0.001) and clinical stage (P > 0.001)., Conclusions: Significant reduction in plasma levels of selenium is recorded in patients undergoing radiotherapy, suggesting attention to the nutritional status of this micronutrient and other antioxidant agents.

The influence of ionic substituents in calcium phosphates intended for bone and tooth replacement biomedical applications is an important research topic, owing to the essential roles played by trace elements in biological processes. The present study investigates the mechanical and biological evaluation of ionic doped hydroxyapatite/β-tricalcium phosphate mixtures which have been prepared by a simple aqueous precipitation method. Heat treating the resultant calcium phosphates in a carbonated atmosphere led to the formation of ionic doped carbonated hydroxyapatite/β-tricalcium phosphate mixtures containing the essential ions of biological apatite. The structural analysis determined by Rietveld refinement confirmed the presence of hydroxyapatite as the main phase, together with a considerable amount of β-tricalcium phosphate. Such phase assemblage is essentially due to the influence of substituted ions during synthesis. The results from mechanical tests proved that carbonate substitutions are detrimental for the mechanical properties of apatite-based ceramics. In vitro proliferation assays of osteoblastic-like cells (MC3T3-E1 cell line) to powders revealed that carbonate incorporation can either delay or accelerate MC3T3 proliferation, although reaching the same proliferation levels as control cells after 2 weeks in culture. Further, the powders enable pre-osteoblastic differentiation in a similar manner to control cells, as indirectly measured by ALP activity and Type-I collagen medium secretion., (Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)