Assessing Bone Health Status and Eggshell Quality of Laying Hens at the End of a Production Cycle in Response to Inclusion of a Hybrid Rye to a Wheat-Corn Diet.

The objective of this study was to evaluate whether there are negative effects of the partial replacement of white corn with rye along with xylanase supplementation on overall bone quality, eggshell mineralization, and mechanical strength in laying hens. From the 26th week of life, ISA Brown laying hens were fed either a wheat-corn diet or a diet containing 25% rye, with or without xylanase. The experimental period lasted for 25 weeks, until birds reached their 50th week of age, after which bone and eggshell quality indices were assessed. Eggshell thickness and eggshell Ca content of eggs from rye-fed hens were improved by xylanase supplementation. No differences in the mechanical properties of the eggshells were observed between treatments, except for the diet-dependent changes in egg deformation. Rye inclusion had no effect on the mechanical properties of bone. Xylanase supplementation, irrespective of the diet, had a positive effect on bone strength and increased tibia Ca content, as well as the content of several microelements. Hence, hybrid rye combined with wheat can replace 25% of corn in layer diets without compromising shell quality or bone mineral content. Xylanase supplementation in these diets is recommended since its inclusion improves both bone strength and quality.

The presence of pathogens and heavy metals in urban peregrine falcons (Falco peregrinus).

BACKGROUND AND AIM: Wild birds raised in urban environments may be exposed to many negative factors, including biological and chemical toxic elements. The aim of the study was to assess the occurrence of bacteria and parasites in wild birds, based on the example of the peregrine falcon (Falco peregrinus) as a potential indicator of bacterial drug resistance genes. Toxicological contamination was also analyzed to determine the impact of urbanized areas on this predatory species, in terms of its health, welfare, and survival in urban environments. MATERIALS AND METHODS: The samples consisted of down feathers and fresh feces obtained from seven falcon chicks (during obligatory veterinary examination) reared in two nests located in the Lublin region (Lublin and Pulawy). Bacteria and parasites were isolated directly from feces by classical microbiological methods, polymerase chain reaction, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MS). The down feathers and feces of birds were used for toxicological testing by plasma inductively coupled plasma MS to assess the concentrations of selected heavy metals (cadmium [Cd], lead [Pb], arsenic [As], zinc [Zn], and copper [Cu]). RESULTS: The study revealed the presence of a diverse microbiome in the falcon chicks, among which Escherichia coli, Enterococcus spp., and Staphylococcus spp. bacteria and parasites of the genus Caryospora were dominant. The presence of drug resistance genes was also confirmed among the pathogens. The toxicological analysis found high concentrations of toxic heavy metals, including Cd, Pb, As, and Zn, in the downy feathers and feces of peregrine chicks. CONCLUSION: Predatory free-living birds living in urban environments not only can be infected with various pathogens but may also show contamination with heavy metals, which could influence their natural resistance, condition, and welfare.

The Concentration of Selected Heavy Metals in Muscles, Liver and Kidneys of Pigs Fed Standard Diets and Diets Containing 60% of New Rye Varieties.

The carry-over of heavy metals from feed to muscles is generally low if animals are fed with a standard diet containing amounts below the maximum permissible levels. However, prolonged exposure to heavy metals can lead to their accumulation in some organs like muscles, liver, and kidneys. This paves the way for human health risks related to the consumption of products of animal-origin. Thus, using feed mixtures with a low level of heavy metals in pig production will contribute to increasing public health and safety and is of environmental concern. The study aimed to assess the impact of the level of some heavy metals (Cd, Pb, Hg, Cu, Zn, Fe and Mn) in standard (control) feed mixtures and in alternative feed mixtures based on maize or new rye varieties (population and hybrid) on the heavy metal concentration in muscles, liver and kidney of fattened pigs at slaughter. While some differences between heavy metals content in examined tissue samples from experimental groups were observed, all of them were in the range of allowable levels according to European Community rules. In conclusion, new rye varieties, especially the hybrid variety, could be an alternative source of cereal grains for pig nutrition.

Modern Hybrid Rye, as an Alternative Energy Source for Broiler Chickens, Improves the Absorption Surface of the Small Intestine Depending on the Intestinal Part and Xylanase Supplementation.

The current study investigated the effects of the inclusion of modern hybrid rye (Brasetto variety) to a corn-wheat-based diet, with or without xylanase, on the absorptive surface of the small intestine of broilers. A total of 224 one-day-old male Ross 308 broiler chicks were randomly divided into four experimental groups with seven replicate cages of eight birds/replicate. A 2 × 2 factorial study design was used, with rye inclusion (0% or 20%) and xylanase supplementation (0 or 200 mg/kg of feed) as factors. Inclusion of rye increased duodenal and ileal crypt depth, villi height, the villus-to-crypt ratio and absorption surface area (p < 0.05), and ileal mucosa thickness and crypt width (p < 0.05). Xylanase supplementation attenuated the effects of rye in the duodenum and ileum and decreased the villi height and villus-to-crypt ratio in the jejunum (p < 0.05). Rye and xylanase had no effect on the spatial distribution of claudin 3 and ZO-1 protein, but xylanase supplementation reduced the amount of claudin 3 in the duodenum and jejunum (p < 0.05). The findings of this study indicate that 20% inclusion of modern hybrid rye to the diets of broilers improved the structure of the duodenum and ileum, but these effects were attenuated by xylanase supplementation.

[Atypical antipsychotics for delirium in hospitalized patients]. / Antipsicóticos atípicos para el tratamiento de delirium en pacientes hospitalizados.

INTRODUCTION: For the pharmacotherapy of delirium in elderly adults who are hospitalized, atypical antipsychotics are used. Currently, there is insufficient evidence on the effectiveness of this treatment in low complexity units. METHODS: We searched in Epistemonikos, the largest database of systematic reviews in health, which is maintained by screening multiple information sources, including MEDLINE/PubMed, EMBASE, Cochrane, among others. We extracted data from the systematic reviews, reanalyzed data of primary studies, conducted a meta-analysis, and generated a summary of findings table using the GRADE approach. RESULTS AND CONCLUSIONS: We identified 13 systematic reviews that included three randomized trials. We concluded that the use of atypical antipsychotics in hospitalized patients likely increases the risk of mortality compared to placebo and could decrease the response rate associated with reducing the risk of adverse effects. Furthermore, using atypical antipsychotics probably results in low or no difference in the severity of delirium.

INTRODUCCIÓN: Para el tratamiento farmacológico de delirium en pacientes adultos mayores hospitalizados se plantea el uso de antipsicóticos atípicos. Actualmente, existe poca evidencia sobre la efectividad y los resultados de estos fármacos en pacientes adultos hospitalizados en unidades de baja complejidad. MÉTODOS: Realizamos una búsqueda en Epistemonikos, la mayor base de datos de revisiones sistemáticas en salud, la cual es mantenida mediante el tamizaje de múltiples fuentes de información, incluyendo MEDLINE/PubMed, EMBASE y Cochrane. Extrajimos los datos desde las revisiones sistemáticas identificadas, analizamos los datos de los estudios primarios, realizamos un metanálisis y preparamos una tabla de resumen de los resultados utilizando el método GRADE. RESULTADOS Y CONCLUSIONES: Identificamos 13 revisiones sistemáticas que en conjunto incluyeron tres estudios primarios, de los cuales todos corresponden a ensayos aleatorizados. Concluimos que el uso de antipsicóticos atípicos en pacientes hospitalizados probablemente aumenta el riesgo de mortalidad en comparación con el placebo, y podría disminuir la tasa de respuesta asociado a disminuir el riesgo de efectos adversos. Además, probablemente resulta en poca o nula diferencia en la severidad del delirium.

A Matter of Degrees: A Systematic Review of the Ergogenic Effect of Pre-Cooling in Highly Trained Athletes.

The current systematic review evaluated the effects of different pre-cooling techniques on sports performance in highly-trained athletes under high temperature conditions. PubMed/MEDLINE, EMBASE, Web of Science, CENTRAL, Scopus, and SPORTDiscus databases were searched from inception to December 2019. Studies performing pre-cooling interventions in non-acclimatized highly-trained athletes (>55 mL/kg/min of maximal oxygen consumption) under heat conditions (≥30 °C) were included. The searched reported 26 articles. Pre-cooling techniques can be external (exposure to ice water, cold packs, or cooling clothes), internal (intake of cold water or ice), or mixed. Cooling prior to exercise concluded increases in distance covered (1.5-13.1%), mean power output (0.9-6.9%), time to exhaustion (19-31.9%), work (0.1-8.5%), and mean peak torque (10.4-22.6%), as well as reductions in completion time (0.6-6.5%). Mixed strategies followed by cold water immersion seem to be the most effective techniques, being directly related with the duration of cooling and showing the major effects in prolonged exercise protocols. The present review showed that pre-cooling methods are an effective strategy to increase sports performance in hot environments. This improvement is associated with the body surface exposed and its sensibility, as well as the time of application, obtaining the best results in prolonged physical exercise protocols.

The effect of cadmium exposition on the structure and mechanical properties of rat incisors.

Alterations in the structure and mechanical properties of teeth in adult Wistar rats exposed to cadmium were investigated. Analyses were conducted on two sets of incisors from female and male specimens, that were intoxicated with cadmium (n = 12) or belonged to the control (n = 12). The cadmium group was administered with CdCl2 dissolved in drinking water with a dose of 4mg/kgbw for 10 weeks. The oral intake of cadmium by adult rats led to the range of structural changes in enamel morphology and its mechanical features. A significant increase of cadmium levels in the teeth in comparison to the control, a slight shift in the colour and reduction of pigmented enamel length, higher surface irregularity, a decrease of hydroxyapatite crystals size in the c-axis and simultaneous increase in pigmented enamel hardness were observed. The extent of these changes was sex-dependent and was more pronounced in males.

Pancreatic-like enzymes of microbial origin restore growth and normalize lipid absorption in a pig model with exocrine pancreatic insufficiency.

INTRODUCTION: The standard therapy for exocrine pancreatic insufficiency (EPI) is porcine-derived pancreatic enzyme replacement therapy (PERT). In the present study we tested a new approach with a mixture of pancreatic-like enzymes of microbial origin (PLEM) in a 1-week efficacy study in EPI pigs. In addition to the conventionally used coefficient of fat and nitrogen absorption (CFA and CNA), parameters that more accurately reflect the nutritional and health status, such as changes in the lipemic index (LI), plasma triglyceride (TG) and non-esterified fatty acid (NEFA) levels, and somatic growth, were determined. MATERIAL AND METHODS: A PLEM dose containing 120 000 active lipase units, 80 000 active protease units and 12 000 active amylase units (all from Sigma, St. Louis, MO) was given as a powder, twice daily with a meal (40 g fat/meal) to 8 EPI pigs for 7 days. Ten healthy pigs were used as a comparator. RESULTS: The PLEM enhanced fat and protein digestion, and reversed growth impairment in EPI pigs. With treatment, CFA and CNA increased by 59% and 43% (p < 0.05), respectively. Although fat and protein absorption were lower than in the comparator, the postprandial blood lipid profile was normal as in healthy pigs. The mucosal thickness significantly increased by 27%, 50% and 26%, in the proximal, middle, and distal jejunum (p < 0.05) with treatment and resembled that of healthy animals. CONCLUSIONS: Pancreatic-like enzymes of microbial origin supported somatic growth and normalized the postprandial lipid profile. As a measure of efficacy, postprandial LI, TG and NEFA are viable endpoints to be explored in human trials.

Stigma-Stop: A Serious Game against the Stigma toward Mental Health in Educational Settings.

This paper presents the results from the application of a serious game called Stigma-Stop among a group of high school students with the aim of reducing the stigma toward mental illnesses. The video game features characters with various mental disorders (schizophrenia, depression, bipolar disorder, and panic disorder with agoraphobia) and provides information about these problems. Additionally, the game asks players about whether they have ever felt the same as the characters, if they believe the characters are psychologically well, and if they think they could help these individuals. Similarly, a variety of reactions are provided for players to choose from when they encounter the characters with these problems. A total of 552 students between the ages of 14 and 18 participated in the study, and they were randomly assigned to either the experimental group, which used Stigma-Stop, or the control group, which utilized a video game completely unrelated to mental health. Both video games were used for similar lengths of time. Following the application of Stigma-Stop, a statistically significant decrease was obtained in levels of stigma toward schizophrenia, both in terms of stereotypes and, to a greater extent, its potential dangerousness. However, this was not the case in the control group. Results thus demonstrate the video game's usefulness toward eradicating erroneous notions about serious mental disorders like schizophrenia.

Intestinal microbiota - a key to understanding the pathophysiology of anorexia nervosa? / Mikrobiota jelitowa ­ klucz do zrozumienia patofizjologii jadlowstretu psychicznego?

Anorexia nervosa (AN) is a psychiatric disorder related to very serious consequences for physical and mental health of patients. Due to a complex clinical picture, which consists of anumber of somatic and mental symptoms, AN remains a serious problem of modern medicine and encourages the search for possible causes of the illness and new, more effective therapies. The recent reports emphasize the role of the intestinal microbiota in regulation of body weight. In this light, the hypothesis that in AN patients there is a significant imbalance of the intestinal microbiota, which contributes to the pathogenesis of the illness, seems interesting. The results of the latest research suggest that abnormal composition of the intestinal microbiota may be an important factor supporting cachexia of AN patients. Detailed analyzes of the composition of the microbiota characteristic for anorexia nervosa could be useful in developing new methods for monitoring and treatment of this illness. This paper aims to present the current state of the art about the role of the intestinal microbiota in the pathogenesis, course and treatment of AN.

Development of a rapid, robust, and universal picogreen-based method to titer adeno-associated vectors.

Recombinant adeno-associated viruses (rAAVs) are promising vectors in preclinical and clinical assays for the treatment of diseases with gene therapy strategies. Recent technological advances in amplification and purification have allowed the production of highly purified rAAV vector preparations. Although quantitative polymerase chain reaction (qPCR) is the current method of choice for titrating rAAV genomes, it shows high variability. In this work, we report a rapid and robust rAAV titration method based on the quantitation of encapsidated DNA with the fluorescent dye PicoGreen®. This method allows detection from 3×10(10) viral genome/ml up to 2.4×10(13) viral genome/ml in a linear range. Contrasted with dot blot or qPCR, the PicoGreen-based assay has less intra- and interassay variability. Moreover, quantitation is rapid, does not require specific primers or probes, and is independent of the rAAV pseudotype analyzed. In summary, development of this universal rAAV-titering method may have substantive implications in rAAV technology.

Construction, production, and purification of recombinant adenovirus vectors.

Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. In this chapter, a standard procedure for their generation and small-scale production is described. Homologous recombination in E. coli between shuttle plasmids and full-length adenovirus backbones (E1-deleted) is used for the generation of recombinant adenoviral vectors genomes. The adenovirus genomes are then analyzed to confirm their identity and integrity, and further linearized and transfected to generate a recombinant adenoviral vector in permissive human cells. These vectors are then purified by two sequential CsCl gradient centrifugations and subjected to a chromatography step in order to eliminate the CsCl and exchange buffers. Finally, the viral stock is characterized through the quantification of its viral particle content and its infectivity.

Canine adenovirus downstream processing protocol.

Adenovirus vectors are efficient gene delivery tools. A major caveat with vectors derived from common human adenovirus serotypes is that most adults are likely to have been exposed to the wild-type virus and exhibit active immunity against the vectors. This preexisting immunity limits their clinical success. Strategies to circumvent this problem include the use of nonhuman adenovirus vectors. Vectors derived from canine adenovirus type 2 (CAV-2) are among the best-studied representatives. CAV-2 vectors are particularly attractive for the treatment of neurodegenerative disorders. In addition, CAV-2 vectors have shown great promise as oncolytic agents in virotherapy approaches and as vectors for recombinant vaccines. The rising interest in CAV-2 vectors calls for the development of scalable GMP compliant production and purification strategies. A detailed protocol describing a complete scalable downstream processing strategy for CAV-2 vectors is reported here. Clarification of CAV-2 particles is achieved by microfiltration. CAV-2 particles are subsequently concentrated and partially purified by ultrafiltration-diafiltration. A Benzonase(®) digestion step is carried out between ultrafiltration and diafiltration operations to eliminate contaminating nucleic acids. Chromatography purification is accomplished in two consecutive steps. CAV-2 particles are first captured and concentrated on a propyl hydrophobic interaction chromatography column followed by a polishing step using DEAE anion exchange monoliths. Using this protocol, high-quality CAV-2 vector preparations containing low levels of contamination with empty viral capsids and other inactive vector forms are typically obtained. The complete process yield was estimated to be 38-45 %.

Immunolocalization of NOS, VIP, galanin and SP in the small intestine of suckling pigs treated with red kidney bean (Phaseolus vulgaris) lectin.

Lectins belong to a family of glycoproteins that can act both beneficially and detrimentally on the morphology of the small intestine. The aim of the study was to determine whether experimental treatment with red kidney bean (Phaseolus vulgaris) lectin influences the chemical code of the small intestine nervous system of suckling pigs. The immunolocalization sites of vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), substance P (SP) and galanin were determined in control and lectin-treated animals. In all segments of the small intestine (duodenum, jejunum, ileum), the subpopulations of VIP-, NOS-, SP- and galanin-immunoreactive (IR) myenteric neurons were unchanged. After lectin stimulation, increased proportions of NOS-IR and decreased numbers of VIP-IR submucous neurons/mucosa innervating nerve fibers were observed in the duodenum, jejunum and ileum. In lectin-treated animals down-regulation of submucous neurons expressing SP and up-regulation of galanin-IR submucous neurons were seen in the duodenum and jejunum (but not in the ileum). The distribution patterns of NOS-IR, galanin-IR and SP-IR nerve fibers supplying the duodenum, jejunum and ileum of the lectin-treated animals showed no substantial differences in relation to control piglets. We conclude that exposure to red kidney bean (P. vulgaris) lectin substantially changes the chemical content of VIP, NOS, SP and galanin in submucous neurons of the small intestine. These results are in line with previous findings outlining the key role(s) of these substances in enteric neuroplasticity processes and may constitute the basis for further functional studies on maturation of the gut.

High concentration of kynurenic acid in bile and pancreatic juice.

Kynurenic acid (KYNA) is an agonist of the G-protein-coupled receptor GPR35, which is predominantly expressed in gastrointestinal tissues. The aim of this study was to determine the content of KYNA in gastric juice, bile and pancreatic juice and intestinal content. KYNA was determined by means of high performance liquid chromatography. The mean concentrations of KYNA in human gastric juice is 9.91 +/- 0.71 nM in contrast to human bile (832.5 +/- 204.1 and 306.8 +/- 35.2 nM) obtained from patients with cholecystolithiasis and obstructive jaundice, respectively. In pigs, the KYNA levels in bile and pancreatic juice are 1,113.3 +/- 63.34 and 757.0 +/- 394.4 nM, respectively. The KYNA concentration increases along the digestive system, reaching 1,638 nM in the colon content. We suggest that the liver and pancreas affect the content of kynurenic acid in the lumen of the digestive tract.

Effect of the electrical currents generated by the intestinal smooth muscle layers on pancreatic enzyme activity: an in vitro study.

Gut enzymes in the small intestine are exposed to extremely low electrical currents (ELEC) generated by the smooth muscle. In the present study, the in vitro tests were undertaken to evaluate the effect of these electric currents on the activity of the proteolytic pancreatic digestive enzymes. A simulator generating the typical electrical activity of pig gut was used for these studies. The electric current emitted by the simulator was transmitted to the samples, containing enzyme and its substrate, using platinum plate electrodes. All samples were incubated at 37 degrees C for 1 h. The changes in optical density, corresponding to enzyme activity, in samples stimulated for 1 h with ELEC was compared with that not exposed to ELEC. The obtained results show that the electrical current with the characteristics of the myoelectrical migrating complex (MMC) has an influence on pancreatic enzyme activity. Increased endopeptidase and reduced exopeptidase activity was noticed in samples treated with ELEC. This observation can be of important as analyzed factors which can alter enzymatic activity of the gut, can thus also affect feed/food digestibility.

The effect of pancreatic and biliary depletion on the in vivo pharmacokinetics of digoxin in pigs.

Several transporter systems in the liver and intestine are known to change their expression and function during cholestatic disease states. The objective of the present in vivo study was to investigate the effect of biliary depletion, as a method to mimic cholestasis, on the bioavailability and disposition of digoxin in biliary and pancreatic duct cannulated pigs. The study was divided in two parts. In the first part, a solution of 10 microg/kg digoxin was administered intravenously to the cannulated pigs with intact enterohepatic circulation (Control) and during depletion of the bile and pancreatic juice. In the second part, the same dose of digoxin was administered intraduodenally with intact enterohepatic circulation (Control) and during depletion of either bile or pancreatic juice or both. Biliary depletion decreased the flow of bile and pancreas juice as well as the amount of digoxin appearing in the bile. Deprivation of both bile and pancreas juice significantly increased the bioavailability of digoxin, the plasma AUC after enteral administration increased from 17.6+/-4.2 nmol/lh (Control) to 29.6+/-8.3 nmol/lh (P<0.05). The biliary clearance decreased significantly, from 0.22+/-0.11 l/h/kg (Control) to 0.04+/-0.03 l/h/kg during pancreatic and biliary depletion (P<0.05). There was a significant decrease in elimination half-life (P<0.05) and volume of distribution (P<0.01) during the depletion experiments while the systemic clearance remained unchanged. The results clearly suggest that biliary depletion trigger a short-term downregulation, most likely posttranscriptionally mediated, of a sinusoidal uptake transporter in the liver, possibly a pig ortholog of OATP.

The enzyme levels in blood are not affected by oral administration of a pancreatic enzyme preparation (Creon 10,000) in pancreas-insufficient pigs.

After oral intake, small amounts of intact protein may be absorbed into the blood circulation. The current study investigated whether orally administered pancreatic enzymes were absorbed from the intestine. The study included 28 pigs; 3 control pigs with intact pancreatic function and 25 pigs that were made exocrine pancreas insufficient by duct ligation (20 pigs) or total pancreatectomy (5 pigs). The pigs received a pancreatic enzyme preparation (0, 2, 4, or 8 g of Creon 10,000) together with the feed. The blood plasma was analyzed for pancreatic lipase activity with a [3H]-triolein substrate assay, while (pro)colipase and cationic trypsin(ogen) levels were measured with enzyme-linked immunosorbent assay (ELISA). Administration of Creon (0-8 g) caused no significant changes in plasma (pro)colipase or cationic trypsin(ogen) levels. Lipase activity peaks in plasma samples were found, but they did not correspond to the administration of Creon. The potential source of these plasma lipase activity peaks is discussed. The results showed no absorption into blood of pancreatic enzymes after oral administration (0, 2, 4, or 8 g of Creon mixed with 100 g of feed) to pancreas-insufficient pigs.