Dietary supplementation with canthaxanthin and 25-hydroxycholecalciferol on the incubation performance and fertility of European quail breeders.

This study assessed the effects of combined supplementation with canthaxanthin (Cx) and 25-hydroxycholecalciferol (25-OH-D3) on incubation performance, fertility, and chick quality in European quail breeders. A total of 240 birds were distributed in a completely randomized design with 5 diets and 8 replicates. The animals were fed a basal diet containing 50 µg of vitamin D3 or the basal diet supplemented with 3 ppm Cx and 34.5 µg 25-OH-D3, 6 ppm Cx, and 69 µg 25-OH-D3, 9 ppm Cx and 103.5 µg 25-OH-D3, or 12 ppm Cx and 138 µg 25-OH-D3. Incubation performance was analyzed in 2 periods (32 and 38 wk). Breeders aged 32 wk produced eggs with higher hatchability (P = 0.024), hatchability of fertile eggs (P = 0.026) and lower initial plus mid embryonic mortality (P = 0.021), whereas 38-week-old breeders generated chicks with a higher length at hatching (P < 0.001) and lower final plus pipped embryonic mortality (P = 0.021). In both age groups, Cx + 25-OH-D3 levels had a quadratic effect on egg fertility (P < 0.001), hatchability of total (P < 0.001), and fertile eggs (P < 0.001). The fertility and the number of sperm cells in the perivitelline membrane was analyzed in two periods (26 and 40 wk). A quadratic effect of diet and days after mating on both parameters (P < 0.05) was observed. Eggs from supplementing breeders showed a high fertility (P < 0.001) and sperm cell counts (P < 0.001) for up to 7 and 3 d after mating, respectively, then the control group. Moreover, the supplementation of quail breeder diets with 6 ppm Cx + 69 µg 25-OH-D3 enhances sperm cell longevity in sperm storage tubules, hatchability of total and fertile eggs, fertility, and chick quality, especially in older quail's breeders and reduces embryonic mortality.

Adaptive statistical iterative reconstruction for computed tomography of the spine.

INTRODUCTION: The utility of evaluating a sagittal view of CT of the spine is well-known. In many clinical cases, the sagittal view includes noise generated from surrounding objects and may degrade the image quality. Iterative reconstruction (IR) techniques are useful for noise reduction; however, they can reduce spatial resolution. The aim of this study was to evaluate the effectiveness of the adaptive statistical iterative reconstruction (ASiR) for generating sagittal CT images of the spine when compared to filtered back projection (FBP). METHODS: The image quality of clinical images from 25 patients were subjectively assessed. Three radiologists rated spatial resolution, image noise, and overall image quality using a five-point scale. For objective assessment, z-direction modulation transfer function (z-MTF) was measured using a custom-made phantom. Additionally, z-axis noise power spectrum (z-NPS) was measured using a water phantom. An improved adaptive statistical iterative reconstruction algorithm called ASiR-V was used in this study. Blending levels were 50%, and 100% (ASiR-V50, ASiR-V100, respectively). RESULTS: For subjective assessments, images using ASiR-V100 were determined to have the best overall image quality, despite having received the worst score in the assessment of spatial resolution. For objective assessments, the image using ASiR-V50 and ASiR-V100 curves were slightly degraded in terms of low contrast z-MTF when compared to FBP. CONCLUSION: ASiR-V was effective to improve the image quality when compared with FBP when reviewing sagittal reformats of the spine. IMPLICATIONS FOR PRACTICE: This study suggests that high resolution is not the only thing that is key when reviewing sagittal CT spinal reformats. Such images should be provided as part of routine CT spine protocols, where available.

High leucine levels affecting valine and isoleucine recommendations in low-protein diets for broiler chickens.

Four experiments were conducted to estimate the optimal standardized ileal digestible (SID) level of branched-chain amino acids in low-protein diets during the starter, grower, and finisher periods, using the response surface methodology, and to study their effects on performance and mRNA expression of genes involved in the mechanistic target of rapamycin (mTOR) pathway of broiler chickens from 8 to 21 D of age. In experiments 1, 2, and 3, a total of 1,500 Cobb male broiler chickens were assigned to 15 diets of a central composite rotatable design (CCD) of response surface methodology containing 5 levels of SID Leu, Val, and Ile with 5 replicate pens of 20 birds each. A 3-factor, 5-level CCD platform was used to fit the second-order polynomial equation of broiler performance. In experiment 4, a total of 540 8-day-old Cobb male broiler chickens were distributed in a completely randomized 2 x 3 x 3 factorial arrangement with 2 SID Leu levels (1.28 or 1.83%), 3 SID Val levels (0.65, 0.90, or 1.20%), and 3 SID Ile levels (0.54, 0.79, or 1.09%) for a total of 18 treatments with 5 replicate cages of 6 birds each. High Leu levels impaired (P < 0.05) gain:feed when birds were fed marginal Val or Ile diets. However, gain:feed was restored when both Val and Ile were supplemented to reach adequate or high levels. High Leu levels increased (P < 0.05) mRNA expression of S6K1 and eEF2 genes only in birds fed high Ile levels. Dietary SID Leu, Val, and Ile levels required for gain:feed optimization in low-protein diets were estimated at 1.37, 0.94, and 0.87% during the starter period; 1.23, 0.82, and 0.75% during the grower period; and 1.15, 0.77, and 0.70% during the finisher phase, respectively. Higher Val and Ile levels are required to optimize the effect of Leu supplementation on mRNA expression of mTOR pathway genes in the pectoralis major muscle of broilers from day 1 to 21 after hatch.

Dietary supplementation with canthaxanthin and 25-hydroxycholecalciferol has beneficial effects on bone and oxidative metabolism in European quail breeders.

This study aimed to investigate the effect of supplementation with canthaxanthin (Cx) and 25-hydroxycholecalciferol (25-OH-D3) on the production performance, egg quality, bone mineral content, blood biochemical parameters, and antioxidant status of European quail breeders. Two hundred and forty quail breeders were distributed in a completely randomized design with 5 diets and 8 replicates of 4 females and 2 males were used. All quail breeders received one of 5 diets: basal diet (containing 2,000 IU vitamin D3) or the same diet supplemented with 3 ppm Cx and 34.5 µg 25-OH-D3, 6 ppm Cx and 69 µg 25-OH-D3, 9 ppm Cx and 103.5 µg 25-OH-D3, or 12 ppm Cx and 138 µg 25-OH-D3. Production performance and internal and external egg quality parameters were not influenced by diet. Eggshell dry weight decreased linearly with increasing supplementation levels, and eggshell ash and calcium content increased quadratically. Plasma phosphorus, calcium, and ionic calcium levels in females and plasma ionic calcium levels in males showed a positive quadratic response to dietary supplementation. Femoral and tibiotarsal dry weight and calcium content were influenced by diet. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity in the liver of males and females and in the serum of females showed a positive quadratic relationship with Cx and 25-OH-D3 levels, whereas the malonaldehyde concentration showed a negative quadratic relationship. DPPH scavenging activity in the serum of male quail increased linearly with supplementation. There was a positive quadratic effect on superoxide dismutase gene expression and a positive linear effect on glutathione peroxidase 7 gene expression, suggesting that dietary enrichment with Cx and 25-OH-D3 might help protect spermatozoa against oxidative damage. The dietary supplement was pro-oxidative at high concentrations (above 9 ppm Cx). The results indicate that diets with adequate levels of Cx and 25-OH-D3 have a beneficial effect on calcium and phosphorus metabolism as well as on the antioxidant defense system. We recommend supplementing European quail breeders in the laying period with 6 ppm Cx and 69 µg 25-OH-D3.

Productive performance, bone characteristics, and intestinal morphology of laying hens fed diets formulated with L-glutamic acid.

This study was conducted to evaluate the effects of L-glutamic acid (Glu) on the productive performance, eggshell quality, bone characteristics, and morphologic parameters of laying hens. Two hundred and forty 53-wk-old Hy-Line W36 laying hens were randomly allocated into 5 treatments, 6 replicates, and 8 birds each. The experimental period was 16 wk. The treatments consisted of a Basal diet (Glu calculated at 2.68%) and 4 L-glutamic acid inclusion levels: 2.88, 3.08, 3.28, and 3.48%. Glu inclusion levels in the hens diet had an increasing linear effect (P < 0.05) on eggshell Ca concentration and bone (tibiotarsus) strength index, and a decreasing linear effect (P < 0.05) on the Seedor index. Serum biochemistry results for total and ionic Ca, inorganic P, and alkaline phosphatase were affected by blood collection time (04:00 Pm, 03:00 Am, and 12:00 Pm) but not by the treatments. Jejunum morphometric variables were not influenced by the treatments except for crypt depth, which demonstrated a quadratic effect (P < 0.05). However, proliferating cell nuclear antigen qualitative immunohistochemical analysis of the jejunum showed more positive nuclei in the villus with the addition of Glu. Both with the basal diet and the lowest Glu inclusion treatment, positive nuclei were observed in the crypts and at the base of the villus, while in the treatments with higher Glu levels, positive cells were common all along the villus mucosa, including its extremity. In conclusion, diets formulated with increased levels of Glu had beneficial effects on eggshell Ca concentration, tibiotarsus structure, and proliferative activity of the jejunum of 69-wk-old laying hens. Therefore, dietary L-glutamic acid can be considered an alternative additive to improve bone characteristics in the productive phase of laying hens.

Development and growth of digestive system organs of European and Japanese quail at 14 days post-hatch.

The objective of this study was to evaluate the development and growth of the digestive system organs, from the 11th day of incubation until the 14 d post-hatch in European and Japanese quail. On days 11, 13 and 15 of incubation at hatch and at 4, 7, 10 and 14 d post-hatch, embryos or chicks of European and Japanese quail were analyzed. After 15 d of incubation, samples from stomach and small intestine were analyzed by microscopy. European quail had significantly heavier body weight at 15 d of incubation and after 4 d post-hatch. The digestive system weight progressively increased with age and was similar between European and Japanese quail at 11, 13, and 15 d of incubation and 10 d post-hatch, while relative weight of digestive system was similar between quail type with great values at 4 d post-hatch. For relative weight of the small intestine + pancreas, the weight of the proventriculus and of the gastric ventricle increased significant by among ages analyzed in both types of quail. At hatch, proventriculus had functional secretory cells and mucosa of gastric ventricle had a thin coilin membrane. In small intestine segments, at 15 d of incubation the height of the villi was similar among duodenum, jejunum, and ileum (80 µm). Villi had elongated shape towards the intestinal lumen, covered by enterocytes and dispersed goblet cells with PAS+ and AB+ contend in all segments. The number of goblet cell/villi increased in segments until 7 to 10 d post-hatch. Duodenum increases the villi up to 14 d, while the jejunum and ileum up to 10 and 4 d, respectively. Based on our data in digestive system growth, a shorter period of post-hatch fast and specific diets to quail during first days of growth is recommended to both quail types. It is concluded that the development and growth of different organs of the digestive system up to 14 d of age was similar between European and Japanese quail.

Aberrant accumulation of ErbB4 in progressive supranuclear palsy.

AIMS: The human epidermal growth factor receptor family consists of four members that belong to the ErbB lineage of proteins (ErbB1-4). Neuregulin-1 (NRG1)/ErbB signalling regulates brain development and function. Abnormalities in this signalling have been implicated in the aetiology or development of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis. So, we aimed at investigating whether the expression of NRG1 or ErbB proteins are altered in progressive supranuclear palsy (PSP). METHODS: The brains of 10 PSP and six control patients were investigated by immunohistochemical analysis. RESULTS: Whereas C-terminal ErbB4 immunoreacitivity was partially but distinctly present in the cytoplasm and/or in the nucleus of neurons in control patients, it was rarely observed in the neuronal nuclei in PSP patients. In contrast, neurofibrillary tangles, coiled bodies and threads were robustly immunoreactive for C-terminal ErbB4 in PSP. Double immunofluorescence for C-terminal ErbB4 and phospho-tau revealed co-localization of these proteins within neuronal and glial inclusions. To the contrary, there was no difference in the subcellular localization of NRG1, ErbB1, ErbB2, and N-terminal ErbB4 between control and PSP patients. These proteins were localized in the cytoplasm of neurons. CONCLUSIONS: Our present results suggest that NRG1/ErbB4 signalling could be an important event in the pathogenesis of PSP.

Tibiotarsus bone characteristics and tibial dyschondroplasia incidence of broilers fed diets supplemented with leucine and valine.

Two experiments were conducted to study the effect of standardized ileal digestible (SID) leucine and valine levels on tibiotarsus bone characteristics and the incidence of tibial dyschondroplasia of broilers from day 1 to 21 (Experiment I) and day 21 to 42 post-hatch (Experiment II). Each experimental phase was evaluated independently. In both experiments, a total of 1,500 one-day-old Cobb 500 male broiler chickens were distributed in a completely randomized design 5 × 5 factorial arrangement for a total of 25 treatments. The SID leucine and valine levels were ranged from 10.0 to 19.6 g/kg, and 6.0 to 12.0 g/kg from day 1 to 21 post-hatch, respectively, while day 21 to 42 post-hatch ranged from 10.0 to 18.0 g leucine/kg, and 5.2 to 11.2 g valine/kg. Serum calcium and phosphorus, bone concentrations of calcium, phosphorus and ash, diameter and Seedor index of the tibiotarsus were not affected (p > .05) by the treatments at 21 or 42 days of age. There was an interaction (p ≤.06) between the SID levels of leucine and valine on tibiotarsus breaking strength at 21 days, but not at 42 days of age (p > .05). Tibiotarsus breaking strength was maximized in broilers from day 1 to 21 with the dietary levels of leucine and valine at 14.2 and 9.0 g/kg respectively. Dietary leucine levels reduced linearly (p < .05) the hypertrophic zone of tibiotarsus cartilage at 21 days of age. Therefore, leucine and valine supplementation interact positively on bone strength of broilers from day 1 to 21 post-hatch. Leucine can be a useful amino acid for reducing the hypertrophic cartilage zone in broilers from day 1 to 21, but not from day 21 to 42 post-hatch.

Performance and serum biochemical profile of Japanese quail supplemented with silymarin and contaminated with aflatoxin B1.

The aim of this study was to evaluate the hepatoprotective effect of silymarin in diets contaminated or not with aflatoxin B1 (AFB1) on the productive performance and serum biochemical profile of Japanese quail (Coturnix coturnix japonica) in the laying phase. A total of 240 12-week-old Japanese quail was used in a completely randomized design in a 3 × 2 factorial scheme (additives x contaminated or not with AFB1 - 1,500 µg/kg), totaling 6 treatments and 5 replicates of 8 birds each. The additives used were silymarin (500 g/ton), adsorbent (1 kg/ton), and a control diet (without additive). Of the total aflatoxin content, 84.64% was AFB1; 4.28% was AFB2; 11.07% was AFG1; and AFG2 was not detected. The data were submitted to ANOVA, and means were compared by Tukey's test. There was no interaction (P > 0.05) between the additive and AFB1 on performance parameters. However, the inclusion of AF in diets reduced (P < 0.05) egg weight and feed intake, impairing feed conversion compared to the unchallenged groups. There was an increase (P < 0.05) in blood concentrations of aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), and creatine kinase (CK) in birds challenged with AFB1, regardless of the additive used, characterizing a possible alteration in hepatic metabolism. Serum total protein and globulin levels were reduced (P < 0.05) in birds challenged with toxins. The consumption of diets contaminated with 1,500 µg AFB1/kg altered hepatic function in quail, impairing productive performance and egg weight. The concentrations of silymarin and adsorbent evaluated in this study were not able to mitigate the negative effect of toxins on the metabolism and performance of laying quail.

Supplementation of broiler breeders with fat sources and vitamin e: carry over effect on performance, carcass yield, and meat quality offspring / Suplementação de matrizes de frangos de corte com fontes de gordura e vitamina E: transferência dos efeitos sobre a performance, o rendimento da carcaça e a qualidade da carne da progênie

The aim of this study was to evaluate two sources of oil (soybean and fish) and four additional levels of vitamin E (0, 150, 250 and 350mg/kg diet) in breeder diets between the 42nd and 56th week of age and its effect on performance and meat quality of offspring. The supplementation of fish oil in the maternal diet increased the deposition of DHA and CLA in egg yolk. From 1 to 35 days of age, intermediate levels of vitamin E resulted in lower weight gain and feed intake. To feed conversion, the best result was obtained supplementing the maternal diet with soybean oil, regardless of vitamin inclusion. In addition, the evaluation of meat quality, the fish oil-supplemented-diet resulted in lower moisture loss of the breast fillets. For maternal diet supplemented with fish oil, the inclusion of vitamin E resulted in a linear increase of the pH value of the meat. Greater levels of vitamin E in the maternal diet influenced the functional properties of meat according to the type of oil that was added to the diet. The deposition of different fatty acids in the embryonic egg yolk can directly influence the meat quality of offspring.(AU)

O objetivo deste estudo foi avaliar duas fontes de óleo (soja e peixe) e quatro níveis adicionais de vitamina E (0, 150, 250 e 350mg/kg dieta), em dietas de matrizes entre a 42º e a 56º semana de idade, sobre o desempenho e a qualidade de carne da progênie. A suplementação de óleo de peixe na dieta das matrizes aumentou a deposição de DHA e CLA na gema dos ovos. Entre um e 35 dias de idade, níveis intermediários de vitamina E resultaram em menor ganho de peso e consumo de ração. Para a conversão alimentar, o melhor resultado foi obtido suplementando a dieta materna com óleo de soja, independentemente da inclusão vitamínica. Dietas suplementadas com óleo de peixe resultaram em menor perda de umidade nos filés de peito. Para a dieta materna suplementada com óleo de peixe, a inclusão de vitamina E resultou em um aumento linear do valor de pH da carne. Níveis mais altos de vitamina E na dieta materna influenciaram as propriedades funcionais da carne de acordo com o tipo de óleo adicionado à dieta. A deposição de diferentes ácidos graxos na gema de ovos embrionados pode influenciar diretamente a qualidade da carne da progênie.(AU)

Context-dependent role for chromatin remodeling component PBRM1/BAF180 in clear cell renal cell carcinoma.

A subset of clear cell renal cell carcinoma (ccRCC) tumors exhibit a HIF1A gene mutation, yielding two ccRCC tumor types, H1H2 type expressing both HIF1α and HIF2α, and H2 type expressing HIF2α, but not functional HIF1α protein. However, it is unclear how the H1H2 type ccRCC tumors escape HIF1's tumor-suppressive activity. The polybromo-1 (PBRM1) gene coding for the BAF180 protein, a component of the SWItch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex, is inactivated in 40% ccRCCs, the function and mechanism of BAF180 mutation is unknown. Our previous study indicates that BAF180-containing SWI/SNF chromatin remodeling complex is a co-activator for transcription factor HIF to induce HIF target genes. Thus, our questions are if BAF180 is involved in HIF-mediated hypoxia response and if PBRM1/BAF180 mutation has any association with the HIF1A retention in H1H2 type ccRCC. We report here that BAF180 is mutated in H1H2 ccRCC cell lines and tumors, and BAF180 re-expression in H1H2 ccRCC cell lines reduced cell proliferation/survival, indicating that BAF180 has tumor-suppressive role in these cells. However, BAF180 is expressed in HIF1-deficient H2 ccRCC cell lines and tumors, and BAF180 knockdown in H2 type ccRCC cell lines reduced cell proliferation/survival, indicating that BAF180 has tumor-promoting activity in these cells. In addition, our data show that BAF180 functions as co-activator for HIF1- and HIF2-mediated transcriptional response, and BAF180's tumor-suppressive and -promoting activity in ccRCC cell lines depends on co-expression of HIF1 and HIF2, respectively. Thus, our studies reveal that BAF180 function in ccRCC is context dependent, and that mutation of PBRM1/BAF180 serves as an alternative strategy for ccRCC tumors to reduce HIF1 tumor-suppressive activity in H1H2 ccRCC tumors. Our studies define distinct functional subgroups of ccRCCs based on expression of BAF180, and suggest that BAF180 inhibition may be a novel therapeutic target for patients with H2, but not H1H2, ccRCC tumors.

Evaluation of the effectiveness of the potent bis-quaternary ammonium compound, 4,4'-(α,ω-hexametylenedithio) bis (1-octylpyridinium bromide) (4DTBP-6,8) on Pseudomonas aeruginosa.

AIMS: Quaternary ammonium compounds (QACs), including benzalkonium chloride (BAC) and cetylpyridinium chloride (CPC) are cationic surfactants and have been used widely as general disinfectants in the medical field due to their strong antibacterial effects and low cytotoxicity to human cells. 4,4'-(α,ω-hexametylenedithio) bis (1-octylpyridinium bromide) (4DTBP-6,8) is one of the potent bis-QACs synthesized to improve the antimicrobial activities of mono-QACs such as BAC. This study aimed to assess the effectiveness of 4DTBP-6,8 against Pseudomonas aeruginosa, a prevalent hospital pathogen. METHODS AND RESULTS: The minimum inhibitory concentrations of 4DTBP-6,8, CPC and BAC against P. aeruginosa were measured. 4DTBP-6,8 exhibited strong antibacterial activity. We assessed the bactericidal effects of QACs against P. aeruginosa under certain conditions and their cytotoxicities in human epithelial cells using lactate dehydrogenase (LDH) release. 4DTBP-6,8 exerted excellent bactericidal effects against high concentrations of bacteria, biofilm cells and even in the presence of contaminated proteins. Cellular LDH was not released by the treatment with 4DTBP-6,8. CONCLUSIONS: 4DTBP-6,8 exhibited the strongest bactericidal activity against P. aeruginosa among the three QACs tested without any cytotoxicity. SIGNIFICANCE AND IMPACT OF THE STUDY: The potent bis-QAC, 4DTBP-6,8 has the potential to be an effective disinfectant in preventing hospital infections caused by P. aeruginosa.

Leucine and valine supplementation of low-protein diets for broiler chickens from 21 to 42 days of age.

The objective of this study was to determine the requirements and interactions between the standardized ileal digestible (SID) Leu and Val levels in low-protein diets, and their effects on performance, serum characteristics, carcass yield and diameter of muscle fibers of broiler chickens from d 21 to 42 posthatch. A total of 1,500 21-day-old Cobb 500 male broiler chickens were distributed in a completely randomized design in a 5 × 5 factorial arrangement for a total of 25 treatments with 3 replicates of 20 birds each. Treatments consisted of 5 SID Leu levels (1.0, 1.2, 1.4, 1.6, or 1.8%) and 5 SID Val levels (0.52, 0.67, 0.82, 0.97, or 1.12%). At 42 d of age, there was interaction (P < 0.05) between the SID levels of Leu and Val on feed intake and weight gain. There was a quadratic effect (P < 0.05) of Leu and Val levels on feed conversion, with minimal point estimated at the levels of 1.19 and 0.86%, respectively. Dietary Leu supplementation reduced linearly (P < 0.05) serum concentrations of triglycerides and ß-hydroxybutyrate. Dietary Leu increased (P ≤ 0.05) the fiber diameters of the pectoralis major muscle and breast yield at the levels of 1.24 and 1.13%, respectively, while the thigh yield was improved with the level of 0.71% Val. Abdominal fat decreased linearly (P < 0.05) with increasing levels of dietary Leu and Val. The SID Leu and Val levels needed to optimize weight gain and feed conversion in low-CP diets for broiler chickens from d 21 to 42 posthatch were estimated at 1.15 and 0.86%, and 1.19 and 0.86%, respectively. The supplementation of Leu and Val can reduce the abdominal fat deposition in birds fed low-CP diets during the grower phase. Leu and Val interactions can influence the performance but not the serum characteristics, carcass yield and diameter of muscle fibers of broilers fed low-protein diets. Therefore, it is necessary to consider the dietary Leu content to estimate the ideal level of Val in low-CP diets for optimum broiler performance.

Single stage incubators and Hypercapnia during incubation affect the vascularization of the chorioallantoic membrane in broiler embryos.

Incubation management can have direct effects on neonate health and consequently affect post-hatching development. The effects of incubation in multiple and single stage incubators with different concentrations of CO2 were evaluated in terms of the vessel density in the chorioallantoic membrane, hatching, heart morphology, and body development of the neonate up to the tenth day. A total of 2,520 fertile eggs were used and distributed in a completely randomized design with 4 levels of CO2 in 4 single-stage incubators (4,000; 6,000; 8,000; and 10,000 ppm) and a control treatment based on multiple-stage incubation, totaling 5 treatments. The levels of CO2 were used during the first 10 d of the incubation period, and after this period, all eggs were submitted to the same level of CO2 (4,000 ppm). Eggs that were incubated in multiple-stage incubators presented a lower percentage of vessels in the chorioallantoic membrane, lower yolk absorption by the embryo, wall depth of the right ventricle, and greater humidity losses in the eggs when compared to eggs in the single-stage incubators. The eggs submitted to hypercapnia, between 5,000 and 6,000 ppm of CO2, had a higher percentage of vessels in the chorioallantoic membrane; the embryos originating from these eggs had higher weight, with higher relative weight of the liver. However, the same levels reduced the yolk absorption. Single-stage incubation with moderate levels of hypercapnia is an efficient tool to be adopted by the hatcheries when attempting to improve chick quality.

Caecal microbiota of chickens fed diets containing propolis.

The present study aimed to evaluate the effect of different levels of ethanolic extract of propolis (EEP) and raw propolis (RP) on broiler performance and on selected bacterial groups in caecal microbiota using fluorescent in situ hybridization (FISH) measured by fluorescent activated cell sorting. Two experiments were conducted with 120 male chicks from 1 to 21 days of age for each, raised in cages and distributed in a completely randomized experimental design; there were five replicates with four birds per experimental unit and six treatments for each experiment (trial 1 - EEP - 0, 1000, 2000, 3000, 4000 and 5000 ppm and trial 2 - RP - 0, 100, 200, 300, 400 and 500 ppm). Fluorescent probes were used against the bacterial groups in caecal samples collected at 21 days of age. The data were subjected to one-way anova followed by Tukey's and regression analyses were used to assess the relationship between dietary levels of EEP or RP on performance and intestinal microbiota (p < 0.05). In the trial 1, results showed that the EEP did not cause any significant (p > 0.05) modification in the performance and caecal microbiota. In the trial 2, RP inclusion did not affect the performance but changed the bacterial composition (p < 0.05). Clostridiaceae, Gammaproteobacteria excluding Enterobacteriaceae and Lactobacillus spp. showed a quadratic response (p < 0.05), with the lowest value predicted to occur at 240 ppm, 221 ppm and 213 ppm of RP respectively. The proportion of Bacteroidaceae and Gammaproteobacteria did not differ (p > 0.05) among the experimental groups. The inclusion of ethanolic extract of propolis did not affect the performance and intestinal microbiota, whereas the supplementation of raw propolis modulates the caecal microbiota composition without any effects on chicken performance.

Influence of graded levels of calcium and vitamin K in the diets of laying hens during the growing phase and their effects on the laying phase.

The aim of this study was to evaluate the effect of dietary supplementation of vitamin K (vit K) and Ca in the diets of laying hens on bone and blood parameters and performance during the growing phase and the effects on the laying phase up to 32 weeks. The study utilized 120 Hy-Line W-36 pullets in the growing phase (13-18 weeks), 80 laying hens in the production phase (20-32 weeks), distributed in a completely randomized design in a 2 × 5 factorial arrangement, two levels of Ca (0.8 and 1.4%) and five levels of vit K supplementation (0, 2, 8, 16 and 32 mg/kg) with four experimental units. An experimental diet was fed during the growing phase, and the commercial diet was fed during the production phase and were analysed for biochemistry and bone variables at 18 and 32 weeks. In pullets, a significant interaction of levels of vit K and Ca was observed for total serum calcium (mg/dl), Seedor index, Ca in the bones (%) and medullary bone (%). Pullets at 18 weeks of age, receiving diets with 1.4% Ca, displayed a quadratic effect of vit K with better results for these variables with 17.86, 14.59, 14.48 and 16.80 mg/kg of vit K, respectively. For level 0.8% Ca no effect of vit K was observed. The treatments during the growing phase had no effect on egg production, performance and biochemistry serum until 32 weeks. Medullary bone (%), there was a significant interaction for 1.4% Ca and a quadratic effect for vit K, with greater medullary bone areas observed with 6.09 mg/kg vit K in the diet. Supplementation with vit K in a diet with 1.4% Ca during the growing phase allowed for an increase in the area of medullary bone at 18 weeks, which was maintained until 32 weeks.

Antimelanoma differentiation-associated protein 5 antibody level is a novel tool for monitoring disease activity in rapidly progressive interstitial lung disease with dermatomyositis.

BACKGROUND: Antimelanoma differentiation-associated protein (anti-MDA)5 antibodies are associated with rapidly progressive interstitial lung disease (RP-ILD) in patients with clinically amyopathic dermatomyositis (CADM) or dermatomyositis (DM). OBJECTIVES: We aimed to evaluate the relevance of monitoring anti-MDA5 antibody levels for the management of RP-ILD in patients with CADM or DM. METHODS: Twelve patients with CADM (n = 10) or DM (n = 2) accompanied by RP-ILD were included. Baseline characteristics and outcomes were recorded. Serial measurements of anti-MDA5 antibody levels were measured. All patients were treated with corticosteroids, tacrolimus and intravenous cyclophosphamide. RESULTS: All patients achieved RP-ILD remission after combined immunosuppressive therapy for a mean of 6·8 months, with significant decreases noted in the mean anti-MDA5 antibody levels at remission. Six (50%) patients became anti-MDA5 antibody negative after therapy. After a mean follow-up of 31 months, RP-ILD relapse was observed in four (33%) patients in both the anti-MDA5 antibody sustained positive group and the negative conversion group. However, relapsed patients in the sustained positive group relapsed earlier than those in the negative conversion group. Thus, a decrease in anti-MDA5 antibody levels during remission was associated with longer remission. Relapses were associated with a reincrease of anti-MDA5 antibody levels in four of four (100%) patients. In contrast, none of the patients without reincrease in anti-MDA5 antibody exhibited symptoms of relapse during follow-up. Therefore, reincrease in anti-MDA5 antibody levels was associated with relapse. CONCLUSIONS: The anti-MDA5 antibody level is a novel parameter for monitoring and a good predictor of RP-ILD relapse in patients with CADM or DM.

Abnormalities in chromosome 6q24 as a cause of early-onset, non-obese, non-autoimmune diabetes mellitus without history of neonatal diabetes.

AIMS: Abnormalities in the imprinted locus on chromosome 6q24 are the most common causes of transient neonatal diabetes mellitus (6q24-related transient neonatal diabetes). 6q24-Related transient neonatal diabetes is characterized by the patient being small-for-gestational age, diabetes mellitus at birth, spontaneous remission within the first few months and frequent recurrence of diabetes after childhood. However, it is not clear whether individuals with 6q24 abnormalities invariably develop transient neonatal diabetes. This study explored the possibility that 6q24 abnormalities might cause early-onset, non-autoimmune diabetes without transient neonatal diabetes. METHODS: The 6q24 imprinted locus was screened for abnormalities in 113 Japanese patients with early-onset, non-obese, non-autoimmune diabetes mellitus who tested negative for mutations in the common maturation-onset diabetes of the young (MODY) genes and without a history of transient neonatal diabetes. Positive patients were further analysed by combined loss of heterozygosity / comparative genomic hybridization analysis and by microsatellite analysis. Detailed clinical data were collected through the medical records of the treating hospitals. RESULTS: Three patients with paternal uniparental isodisomy of chromosome 6q24 were identified. None presented with hyperglycaemia in the neonatal period. Characteristically, these patients were born small-for-gestational age, representing 27.2% of the 11 patients whose birth weight standard deviation score (SDS) for gestational age was below -2.0. CONCLUSIONS: Abnormalities in the imprinted locus on chromosome 6q24 do not necessarily cause transient neonatal diabetes. Non-penetrant 6q24-related diabetes could be an underestimated cause of early-onset, non-autoimmune diabetes in patients who are not obese and born small-for-gestational age.

Drought impact on forest carbon dynamics and fluxes in Amazonia.

In 2005 and 2010 the Amazon basin experienced two strong droughts, driven by shifts in the tropical hydrological regime possibly associated with global climate change, as predicted by some global models. Tree mortality increased after the 2005 drought, and regional atmospheric inversion modelling showed basin-wide decreases in CO2 uptake in 2010 compared with 2011 (ref. 5). But the response of tropical forest carbon cycling to these droughts is not fully understood and there has been no detailed multi-site investigation in situ. Here we use several years of data from a network of thirteen 1-ha forest plots spread throughout South America, where each component of net primary production (NPP), autotrophic respiration and heterotrophic respiration is measured separately, to develop a better mechanistic understanding of the impact of the 2010 drought on the Amazon forest. We find that total NPP remained constant throughout the drought. However, towards the end of the drought, autotrophic respiration, especially in roots and stems, declined significantly compared with measurements in 2009 made in the absence of drought, with extended decreases in autotrophic respiration in the three driest plots. In the year after the drought, total NPP remained constant but the allocation of carbon shifted towards canopy NPP and away from fine-root NPP. Both leaf-level and plot-level measurements indicate that severe drought suppresses photosynthesis. Scaling these measurements to the entire Amazon basin with rainfall data, we estimate that drought suppressed Amazon-wide photosynthesis in 2010 by 0.38 petagrams of carbon (0.23-0.53 petagrams of carbon). Overall, we find that during this drought, instead of reducing total NPP, trees prioritized growth by reducing autotrophic respiration that was unrelated to growth. This suggests that trees decrease investment in tissue maintenance and defence, in line with eco-evolutionary theories that trees are competitively disadvantaged in the absence of growth. We propose that weakened maintenance and defence investment may, in turn, cause the increase in post-drought tree mortality observed at our plots.

Long-term decline of the Amazon carbon sink.

Atmospheric carbon dioxide records indicate that the land surface has acted as a strong global carbon sink over recent decades, with a substantial fraction of this sink probably located in the tropics, particularly in the Amazon. Nevertheless, it is unclear how the terrestrial carbon sink will evolve as climate and atmospheric composition continue to change. Here we analyse the historical evolution of the biomass dynamics of the Amazon rainforest over three decades using a distributed network of 321 plots. While this analysis confirms that Amazon forests have acted as a long-term net biomass sink, we find a long-term decreasing trend of carbon accumulation. Rates of net increase in above-ground biomass declined by one-third during the past decade compared to the 1990s. This is a consequence of growth rate increases levelling off recently, while biomass mortality persistently increased throughout, leading to a shortening of carbon residence times. Potential drivers for the mortality increase include greater climate variability, and feedbacks of faster growth on mortality, resulting in shortened tree longevity. The observed decline of the Amazon sink diverges markedly from the recent increase in terrestrial carbon uptake at the global scale, and is contrary to expectations based on models.