Dietary Hydroxytyrosol Supplementation on Growth Performance, Gut Morphometry, and Oxidative and Inflammatory Status in LPS-Challenged Broilers.

This study assessed the effects of hydroxytyrosol (HT) on 8- to 20-day-old broilers challenged with lipopolysaccharide (LPS); 180 Cobb500™ male chicks were randomly assigned to 3 treatment groups, each comprising 10 replicates with 6 birds per replicate. Treatments included a control diet (CON), CON with LPS administration, and CON + LPS supplemented with 10 mg of HT/kg of feed. LPS was administered intraperitoneally on days 14, 16, 18, and 20. Body weight (BW), body weight gain (BWG), and the feed conversion ratio (FCR) were measured. On day 20, ten birds per treatment were slaughtered for analysis. Bursa, spleen, and liver were collected, and their respective relative weight was determined. The jejunum was destined for morphological analyses of villus height (VH), crypt depth (CD), and their ratio (VH:CD), and for mRNA expression of nuclear factor kappa B (NF-κB), catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), and interleukins 10 (IL-10), 1 beta (IL-1ß), and 8 (IL-8). HT improved BW, BWG, and FCR, and reduced crypt depth (CD) while increasing the VH:CD ratio in the jejunum. Moreover, HT downregulated mRNA expression of CAT, GPx, IL-10, and IL-1ß. In conclusion, HT enhances broiler growth performance, mitigates jejunal mucosa damage from LPS, and modulates antioxidant and immune responses.

Research Note: Nitrogen-corrected apparent metabolizable energy and standardized ileal amino acid digestibility determination of high-protein dried distiller's grains and corn bran with solubles for broilers.

Metabolism and digestibility trials were performed on broiler chickens to determine 1) nitrogen-corrected apparent metabolizable energy (AMEn) using total excreta collection and 2) standardized ileal amino acid digestibility (SIAAD) using the ileal digesta collection from high-protein dried distillers' grains (HP-DDG) and corn bran with solubles (CBS). The results of the metabolism trial indicated that AMEn values for HP-DDG and CBS were 3,334 kcal/kg and 2,083 kcal/kg on dry matter (DM) basis, respectively. Concerning the HP-DDG, the digestibility trial resulted in the following SIAAD values and digestible concentrations, respectively: 80.33% and 1.09 for Lys, 85.95% and 1.44 for Met + Cys, 75.58% and 1.24 for Thr, 89.58% and 1.66 for Arg, 84.91% and 1.08 for His, 86.37% and 1.35 for Ile, 90.64% and 4.56 for Leu, 85.76% and 1.80 for Val, and 88.67% and 1.90 for Phe. Concerning the CBS, the following SIAAD values and digestible concentrations were measured, respectively: 79.29% and 0.44 for Lys, 89.57% and 0.31 for Met + Cys, 78.89% and 0.40 for Thr, 92.28% and 0.66 for Arg, 87.48% and 0.36 for His, 93.40% and 0.35 for Ile, 92.27% and 1.01 for Leu, 90.97% and 0.51 for Val, and 88.81% and 0.45 for Phe. The digestibility average of CBS is 88.45% for essential amino acids and 85.21% for nonessential amino acids, whereas the digestibility average of HP-DDG is 85.83% for essential amino acids and 83.83% for nonessential amino acids.

Research Note: The influence of different isoleucine: lysine ratios on the growth performance of broiler chickens fed low-protein diets.

Two trials were carried out to assess the effects of different ratios of standardized ileal digestible isoleucine:lysine (SID Ile:Lys) on the growth performance of broiler chickens fed low-protein diets. A total of 1,320 male chickens were distributed in each trial into 6 treatments, with 10 replicates with 22 birds each. A control diet was formulated that satisfied the nutritional requirements of the broilers, and a low-protein diet was formulated with reduced protein content, meeting broiler nutritional requirements, except for the SID Ile levels. Five SID Ile:Lys ratios (56%, 61%, 66%, 71%, and 76%) were obtained by adding l-isoleucine to the low-protein diet. The body weight (BW), body weight gain (BWG), average daily feed intake (ADFI), and feed conversion ratio (FCR) were evaluated from day 1 to day 21 in trial 1, and from day 22 to day 44 in trial 2. ANOVA was performed on the data, and the treatments were compared to the control group using Dunnett's test (P ≤ 0.05). Regression analyses were performed for modeling the variables assessed and the ratios of SID Ile:Lys. There was no significant difference between the treatments on ADFI of birds (P > 0.05). The BW, BWG, and FCR showed a quadratic effect as the SID Ile:Lys ratio increased in low-protein diets in trials 1 and 2 (P ≤ 0.05). In conclusion, the recommended ratio of SID Ile:Lys in low-protein diets for growth performance is around 66% for broiler chickens from 1 to 21 d old and is around 65% for broiler chickens from 22 to 44 d old.

Effects of Hydroxytyrosol Supplementation on Performance, Fat and Blood Parameters of Broiler Chickens.

The study aimed to evaluate the effects of dietary supplementation of hydroxytyrosol (HT) on performance, fat, and blood parameters of broilers. In total, 960 male chicks were distributed into four treatments groups with 12 replicates with 20 birds per pen, with varying HT levels (0, 5, 10, and 50 mg/kg of feed) added to the basal diet from 1 to 42 days old. Feed intake, body weight gain, and feed conversion ratio were evaluated. Enzymes related to liver injury were evaluated in blood. Fatty acid profile and malondialdehyde (MDA) concentration were determined in the breast meat. Dietary supplementation of HT did not improve broilers' performance (p > 0.05). Birds fed 50 mg HT/kg had lower AST, ALT, and GGT concentrations (p ≤ 0.05), whereas broilers fed 5, 10, and 50 mg HT/kg, had lower TBIL concentrations (p ≤ 0.05). Breast meat of broilers fed 50 mg HT/kg had lower lipid content, saturated fatty acid, unsaturated fatty acids, MDA concentrations (p ≤ 0.05), and polyunsaturated fatty acids (p < 0.0001). In summary, supplementation of 5, 10, and 50 mg HT/kg does not improve the performance of broilers, but the dose of 50 mg HT/kg helps the liver against inflammation and improves fat parameters.

The effects of arginine supplementation through different ratios of arginine:lysine on performance, skin quality and creatine levels of broiler chickens fed diets reduced in protein content.

Two trials were carried out to assess the effects of arginine supplementation through ratios of digestible arginine:lysine on growth performance, skin quality and creatine levels in muscle and serum of broiler chickens fed diets reduced in protein content. A total of 1,540 Cobb500 male chickens were distributed into 7 treatments, with 10 replicates with 22 birds each. The experimental diets were based on corn and soybean meal, and a control diet was formulated to satisfy broiler nutritional requirements. A basal diet with reduced protein content was formulated to meet broiler nutritional requirements, except for SID Arg levels. The experimental diets were obtained by adding L-arginine to basal diets, meeting 6 different SID Arg:Lys ratios (94, 100, 106, 112, 118, and 124%). Body weight, body weight gain, average daily feed intake, and feed conversion ratio were evaluated from 01 to 21 d old (trial 1) and from 22 to 44 d old (trial 2). At 21 and 44 d, in trials 1 and 2, respectively, birds were slaughtered to assess skin thickness (ST), skin strength (SS), creatine level in muscle (CRM) and serum (CRS). Data were subjected to ANOVA, and treatments were compared to the control group by Dunnett's test (P ≤ 0.05). Regression analyses were performed to model the variables assessed and the ratios of SID Arg:Lys. The SID Arg:Lys ratios did not affect ADFI of broilers in both trials (P > 0.05), whereas it linearly increased the BW, BWG, and ST, in both trials (P < 0.001). The FCR of broilers linearly decreased, in trial 1 (P = 0.038) and trial 2 (P < 0.001). The CRM of birds had a linear effect (P < 0.001) in trial 1, and a quadratic effect (P = 0.001) in trial 2. The CRS and SS of broilers linearly increased, in trial 2 (P < 0.001). In conclusion, increasing SID Arg:Lys ratios in diets reduced CP enhanced growth performance, skin quality and CR levels in muscle and serum of broiler chickens from 01 to 21 and 22 to 44 d old.

Somatosensory Electrical Stimulation Does Not Augment Motor Skill Acquisition and Intermanual Transfer in Healthy Young Adults-A Pilot Study.

Sensory input can modify motor function and magnify interlimb transfer. We examined the effects of low-intensity somatosensory electrical stimulation (SES) on motor practice-induced skill acquisition and intermanual transfer. Participants practiced a visuomotor skill for 25 min and received SES to the practice or the transfer arm. Responses to single- and double-pulse transcranial magnetic stimulation were measured in both extensor carpi radialis. SES did not further increase skill acquisition (motor practice with right hand [RMP]: 30.8% and motor practice with right hand + somatosensory electrical stimulation to the right arm [RMP + RSES]: 27.8%) and intermanual transfer (RMP: 13.6% and RMP + RSES: 9.8%) when delivered to the left arm (motor practice with right hand + somatosensory electrical stimulation to the left arm [RMP + LSES]: 44.8% and 18.6%, respectively). Furthermore, transcranial magnetic stimulation measures revealed no changes in either hand. Future studies should systematically manipulate SES parameters to better understand the mechanisms of how SES affords motor learning benefits documented but not studied in patients.

Age-related changes in corticospinal excitability and intracortical inhibition after upper extremity motor learning: a systematic review and meta-analysis.

It is unclear how old age affects the neuronal mechanisms of motor learning. We reviewed the neuronal mechanisms of how healthy old and young adults acquire motor skills as assessed with transcranial magnetic stimulation. Quantitative meta-analyses of 11 studies, involving ballistic and visuomotor tasks performed by upper extremity muscles in 132 healthy old and 128 young adults, revealed that the motor practice-induced increase in corticospinal excitability (CSE) is task-dependent but not age-dependent, with an increase in CSE in both age groups after visuomotor but not ballistic training. In addition, short-interval intracortical inhibition (SICI) is reduced in old but not young adults, but only after visuomotor practice. In addition, correlation analyses in 123 old and 128 young adults showed that the magnitude of motor skill acquisition did not correlate with increases in CSE or decreases in SICI in either age group. Thus, there are subtle age-related differences in use-dependent plasticity but increases in CSE or decreases in SICI are not related to motor skill acquisition in healthy young or old adults.

Neuronal mechanisms of motor learning are age dependent.

There is controversy whether age-related neuroanatomical and neurophysiological changes in the central nervous system affect healthy old adults' abilities to acquire and retain motor skills. We examined the effects of age on motor skill acquisition and retention and potential underlying mechanisms by measuring corticospinal and intracortical excitability, using transcranial magnetic stimulation. Healthy young (n = 24, 22 years) and old (n = 22, 71 years) adults practiced a wrist flexion-extention visuomotor task or only watched the templates as an attentional control for 20 minutes. Old compared with young adults performed less well at baseline. Although the absolute magnitude of skill acquisition and retention was similar in the 2 age groups (age × intervention × time, p = 0.425), a comparison of baseline-similar age sub-groups revealed impaired skill acquisition but not retention in old versus young. Furthermore, the neuronal mechanisms differed as revealed by an opposite direction of associations in the age-groups between relative skill acquisition and intracortical facilitation during the task, and opposite changes during skill retention in corticospinal excitability at rest and during the task and intracortical inhibition during the task.

Assessment of muscle architecture of the biceps femoris and vastus lateralis by ultrasound after a chronic stretching program.

OBJECTIVE: To evaluate the chronic effects of a static stretching program on the muscle architecture of biceps femoris (BF) and vastus lateralis (VL) muscles in ultrasound (US) images. DESIGN: Randomized controlled longitudinal trial. SETTING: Biomechanics Laboratory of Physical Education School of the Army, Rio de Janeiro, Brazil. PARTICIPANTS: The study included 24 healthy and physically active male volunteers (19.05 ± 1.40 years, 1.73 ± 0.07 m, and 73.15 ± 8.33 kg), randomly allocated to 1 of 2 groups: stretching group (SG, n = 12) and control group (n = 12). INTERVENTIONS: The SG was submitted to 3 sets of 30 seconds of static stretching 3 times a week during 8 weeks. MAIN OUTCOME MEASURES: Ultrasound equipment (7.5 MHz) was used for the evaluation of BF and VL muscle architecture variables (pennation angle, fiber length, muscle thickness, and fascicle displacement) before and after training. Knee range of motion (ROM) and isometric flexion and extension torque (TQ) were also measured. RESULTS: There were no significant changes in muscle architecture, TQ, and maximum knee flexion angle (P > 0.05). However, maximum knee extension angle (MEA) increased significantly in the SG (pretraining: 159.37 ± 7.27 degrees and posttraining: 168.9 ± 3.7 degrees; P < 0.05). CONCLUSIONS: Volume or intensity (or both) of the stretching protocol was insufficient to cause structural changes in the VL and BF muscles. The increase in MEA could not be explained by muscle architecture changes. CLINICAL RELEVANCE: To describe changes in the VL and BF muscle tendon unit using US after a long-term stretching program to identify which structures are responsible for ROM increase.

Granulomatous PJP presenting as a solitary lung nodule in an immune competent female.

Pneumocystis jiroveci pneumonia (PJP) opportunistically targets immunosuppressed patients, most notably those with advanced HIV/AIDS. Radiologically, PJP typically appears as bilateral diffuse pulmonary infiltrates. Herein an unusual case of an immunocompetent woman developing granulomatous PJP in the absence of evident risk factors is described. PJP may be an under-recognized cause of pulmonary nodules in immune competent individuals.

Reliability of the rectus femoris muscle cross-sectional area measurements by ultrasonography.

The skeletal muscle system can adapt to an external stimulus from either physiological or pathological conditions. This plasticity is measured by imaging techniques such as magnetic resonance imaging or ultrasound. The anatomical cross-sectional area of a muscle is one of the muscle architecture parameters that relates to the maximum muscle strength. The aim of this study was to determine the reliability of anatomical cross-sectional area rectus femoris measurements, obtained by ultrasound, with two different protocols. Acquisition of four anatomical cross-sectional area images of the right rectus femoris in two distinct regions (15 cm above the patella and 50% of the thigh length) was performed in 2 days, from a group of 15 young healthy subjects. The cross-sectional area of each image was measured five times. The reliability of the anatomical cross-sectional area measures was determined by the coefficient of variation (CV), intraclass correlation coefficient (ICC) and typical error of measurement (TEM). In each protocol, there were no significant differences between the means of anatomical cross-sectional area in measurements, images and days (P>0·05). The CVs were 8·53% and 8·9%, the ICCs 0·88 and 0·87 and the TEMs 65·59 and 94·25 between the 2 days in the regions of 15 cm and 50% of the thigh length, respectively. The average values of the cross-sectional area at 50% of the thigh length were significantly higher than those for at 15 cm above the patella (P<0·001). The measurement of rectus femoris anatomical cross-sectional area by ultrasound proved reliable.

Corticosteroid-induced gene expression in allergen-challenged asthmatic subjects taking inhaled budesonide.

BACKGROUND AND PURPOSE: Inhaled corticosteroids (ICS) are the cornerstone of asthma pharmacotherapy and, acting via the glucocorticoid receptor (GR), reduce inflammatory gene expression. While this is often attributed to a direct inhibitory effect of the GR on inflammatory gene transcription, corticosteroids also induce the expression of anti-inflammatory genes in vitro. As there are no data to support this effect in asthmatic subjects taking ICS, we have assessed whether ICS induce anti-inflammatory gene expression in subjects with atopic asthma. EXPERIMENTAL APPROACH: Bronchial biopsies from allergen-challenged atopic asthmatic subjects taking inhaled budesonide or placebo were subjected to gene expression analysis using real-time reverse transcriptase-PCR for the corticosteroid-inducible genes (official gene symbols with aliases in parentheses): TSC22D3 [glucocorticoid-induced leucine zipper (GILZ)], dual-specificity phosphatase-1 (MAPK phosphatase-1), both anti-inflammatory effectors, and FKBP5 [FK506-binding protein 51 (FKBP51)], a regulator of GR function. Cultured pulmonary epithelial and smooth muscle cells were also treated with corticosteroids before gene expression analysis. KEY RESULTS: Compared with placebo, GILZ and FKBP51 mRNA expression was significantly elevated in budesonide-treated subjects. Budesonide also increased GILZ expression in human epithelial and smooth muscle cells in culture. Immunostaining of bronchial biopsies revealed GILZ expression in the airways epithelium and smooth muscle of asthmatic subjects. CONCLUSIONS AND IMPLICATIONS: Expression of the corticosteroid-induced genes, GILZ and FKBP51, is up-regulated in the airways of allergen-challenged asthmatic subjects taking inhaled budesonide. Consequently, the biological effects of corticosteroid-induced genes should be considered when assessing the actions of ICS. Treatment modalities that increase or decrease GR-dependent transcription may correspondingly affect corticosteroid efficacy.

Effect of freezing of sputum samples on flow cytometric analysis of lymphocyte subsets.

Sputum samples should be processed shortly after induction to prevent cell degradation. For intermediate storage, freezing of homogenised samples or immediate fixation have been shown to be suitable for cytospins. The aim of this study was to investigate whether freezing or immediate fixation of sputum affect the analysis of lymphocyte subsets by flow cytometry. Selected plugs from 24 sputum samples were homogenised. One aliquot was processed immediately and analysed by flow cytometry. A second aliquot was homogenised, frozen at -20 C after addition of dimethylsulfoxide and stored for a median time of 6 days. In six samples a third aliquot was fixed in formalin after induction and stored for up to 72 h before further processing. Compared to immediate processing, percentages of total lymphocytes and T-suppressor cells were elevated after being frozen, with a minor decrease in the T4/T8 ratio. Proportions of total lymphocytes, T-helper and T-suppressor cells correlated between native and frozen samples, intra-class correlation coefficients being 0.74, 0.85 and 0.70, respectively. The formalin-fixed aliquots could not be analysed with the antibodies used. In conclusion, freezing seems to be a suitable technique to store sputum samples for flow cytometry of CD3, CD4 and CD8 lymphocyte subsets. Its effects were minor compared to the variation between subjects.

A method to preserve sputum for delayed examination.

Examination of sputum cell counts is limited by the need to process samples within hours of expectoration. The validity and repeatability of a method to preserve sputum for delayed processing and examination were investigated in this study. Portions of selected sputum from 39 subjects were dispersed with dithiothreitol (routine method), or were fixed in a dithiothreitol-formaldehyde mixture for 48-72 h before dispersal with trypsin (preservation method). Total and differential cell counts using the two methods were compared; within-method repeatability of the preservation method was also examined. The intraclass correlation coefficient (ICC) for total cell counts and percentage of eosinophils, neutrophils and macrophages in sputum processed by the two methods was 0.89, 0.86, 0.91 and 0.90, respectively. Within-method repeatability (ICC) of the preservation method for the same cellular indices was 0.99, 0.94, 0.97 and 0.97, respectively. The interobserver repeatability for eosinophils, neutrophils and macrophages was 0.96, 0.97 and 0.97 using the preservation method, and 0.96, 0.99 and 0.99 using the routine method, respectively. This method of sputum preservation and dispersal is valid, reliable and convenient, and may be used for delayed processing and examination.

Increased detection of interleukin-5 in sputum by addition of protease inhibitors.

The measurement of interleukin (IL)-5 in sputum is problematic, with interfering factors affecting immunoassay. The authors investigated whether sputum proteases could be acting as interfering factors by studying the effect of protease inhibitors (PI) on sputum IL-5 measurement. Induced sputa from 20 subjects with asthma were divided into aliquots, processed with and without protease inhibitors (in low and high concentrations) and the levels of IL-5 (spiked and endogenous) measured by enzyme immunoassay were compared. The concentration of sputum IL-5 was significantly increased by PI, with median (interquartile range) levels processed with no, low and high PI concentrations being 0 (0), 41.8 (75.6) and 66.1 (124.4) pg x mL(-1), respectively. There was also a significant increase in percentage recovery of spiked IL-5. Although high concentrations of PI reduced cell viability, there was no effect on total or differential cell counts and low concentrations of PI had no effect on cell counts or viability. Levels of endogenous interleukin-5 in sputum of asthmatic subjects can be significantly increased by the addition of protease inhibitors, and samples which would be regarded as negative for interleukin-5 without protease inhibitors may instead have considerable amounts of interleukin-5 detected.

A tale of three blind men on the proper subject matter of clinical science and practice: commentary on Plaud's behaviorism vs. Ilardi and Feldman's cognitive neuroscience.

Plaud (J Clin Psychol 57, 1089-1102, 1109-1111, 1119-1120) and Ilardi and Feldman (J Clin Psychol 57, 1067-1088, 1103-1107, 1113-1117, 1121-1124) argue for two very different approaches to clinical science and practice (i.e., behavior analysis and cognitive neuroscience, respectively). We comment on the assets and liabilities of both perspectives as presented and attempt to achieve some semblance of balance between the three protagonists embroiled in this current debate. The vision of clinical science we articulate is more ecumenical and evolutionary, rather than paradigmatic and revolutionary. As we see it, the problem clinical psychology faces is much larger than the authors let on; namely, how best to make clinical science meaningful and relevant to practitioners, consumers, the general public, and the behavioral health-care community. Clinical psychology's immediate internal problem is not pluralism with regard to subject matter, worldview, methodology, or school of thought, but pluralism in clinical psychologists' adherence to a scientific epistemology as the only legitimate form of clinical psychology. On this latter point, we still have a very long way to go.

Phospholipase C-gamma mediates the hydrolysis of phosphatidylinositol, but not of phosphatidylinositol 4,5-bisphoshate, in carbamylcholine-stimulated islets of langerhans.

In pancreatic islets the activation of phospholipase C (PLC) by the muscarinic receptor agonist carbamyolcholine (carbachol) results in the hydrolysis of both phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)) and phosphatidylinositol (PtdIns). Here we tested the hypothesis that PtdIns hydrolysis is mediated by PLCgamma1, which is known to be regulated by activation of tyrosine kinases and PtdIns 3-kinase. PtdIns breakdown was more sensitive than that of PtdInsP(2) to the tyrosine kinase inhibitor, genistein. Conversely, the tyrosine phosphatase inhibitor, vanadate, alone promoted PtdIns hydrolysis and acted non-additively with carbachol. Vanadate did not stimulate PtdInsP(2) breakdown. Carbachol also stimulated a rapid (maximal at 1-2 min) tyrosine phosphorylation of several islet proteins, although not of PLCgamma1 itself. Two structurally unrelated inhibitors of PtdIns 3-kinase, wortmannin and LY294002, more effectively attenuated the hyrolysis of PtdIns compared with PtdInsP(2). Adenovirally mediated overexpression of PLCgamma1 significantly increased carbachol-stimulated PtdIns hydrolysis without affecting that of PtdInsP(2). Conversely overexpression of PLCbeta1 up-regulated the PtdInsP(2), but not PtdIns, response. These results indicate that the hydrolysis of PtdIns and PtdInsP(2) are independently regulated in pancreatic islets and that PLCgamma1 selectively mediates the breakdown of PtdIns. The activation mechanism of PLCgamma involves tyrosine phosphorylation (but not of PLCgamma directly) and PtdIns 3-kinase. Our findings point to a novel bifurcation of signaling pathways downstream of muscarinic receptors and suggest that hydrolysis of PtdIns and PtdInsP(2) might serve different physiological ends.