Dexamethasone and prednisolone treatment in beef cattle: influence on glycogen deposition and gene expression in the liver.

The illegal administration of glucocorticoids in livestock is problematic and identification of pathways in which these hormones are involved is critically important, and new direct or indirect biomarkers should be identified. In this work, glucocorticoid transcriptional effects on some genes involved in the glucose metabolism were studied in the bovine liver. This study was conducted on adult Charolais male cattle treated with long-term low dose dexamethasone or prednisolone. Gene expression analysis was conducted in the liver by qPCR, and the geNorm algorithm was applied to select optimal reference genes. In line with the literature, a significant overexpression of genes involved in the gluconeogenic pathway and glycogen synthesis was detected in the liver of dexamethasone-treated animals, but histological and biochemical examination showed hepatocyte glycogen depletion particularly in dexamethasone-treated animals. It possible to hypothesize that glucocorticoids or adrenal insufficiency due to glucocorticoids withdrawal inhibit the enzymatic activity of glycogen synthase and/or induce glycogen autophagy in bovine liver. In fact, markers of glycophagy as starch-binding domain-containing protein 1 and γ-aminobutyric acid receptor-associated protein-like 1 mRNAs were upregulated in the liver by glucocorticoids treatment. Furthermore, glycogen synthase kinase-3 beta gene was significantly overexpressed in dexamethasone-treated animals, and this protein is also implicated in liver autophagy modulation and glycogen synthesis inhibition. These results showed that glucocorticoids likley have dual roles in hepatic glycogen metabolism of cattle, and investigation of these pathways could help find treatment biomarkers.

Plasmodium spp. In a captive raptor collection of a Safaripark in northwest Italy.

Blood parasites infect all vertebrates (Clayton and Moore 1997). Avian malaria parasites (Plasmodium spp., Plasmodiidae) are cosmopolitan in their distribution and are responsible for severe diseases in domestic and wild birds.In September 2009, nine raptorial birds that either arrived recently or were maintained as permanent residents at the Safaripark Pombia (northwest Italy) showed loss of stamina, developing listlessness, anorexia and regurgitation. Within one month three animals died and were necropsied.Following the diagnosis of Plasmodium infection all other raptorial birds were treated: clinical improvement was observed in all birds, and blood smears made after one month resulted negative for parasites.

Ameloblastic fibroma in an alpine chamois (Rupicapra rupicapra).

Spontaneous odontogenic tumors are neoplasms characterized by a mixed odontogenic ectomesenchymal and odontogenic epithelial origin; they are rare in both humans and animals. A 3-year-old male Alpine Chamois (Rupicapra rupicapra) was found dead in north-west Italy, and was referred for the necropsy to the Department of Veterinary Sciences of the University of Turin (Italy). At the external examination a 10 × 8 cm, exophytic, red-pink, smooth, firm and ulcerated mass was observed on the inferior lip. Histologically the tumor was characterized by spindle shaped cells arranged in bundles in an abundant hyaline matrix. Multifocal and rare chords of odontogenic epithelium mixed with rare melanocytes that penetrate the neoplasia were visible. Immunohistochemistry showed a clear cytokeratin positivity of epithelial clusters. Macroscopical, histological and immunohistochemical findings were consistent with a diagnosis of locally infiltrative ameloblastic fibroma. To our best knowledge, this is the first report of this tumor in a wild ungulate and in Alpine Chamois.

Heart pathologies in dolphins stranded along the northwestern Italian coast.

Nine striped dolphins Stenella coeruleoalba and 1 bottlenose dolphin Tursiops truncatus stranded along the Ligurian Sea coast of Italy were necropsied between February 2011 and April 2012. Macroscopic and histological findings were observed in the hearts of all animals and included saccular aneurysms of the pulmonary trunk (n = 3), cirsoid aneurysms (n = 1), right ventricular dilation (n = 1) associated with hypoplasia of the tricuspid chordae (n = 1), valvular fibrosis (n = 3), mitral leaflet thickening (n = 1), left ventricular hypertrophy (n = 1), lymphocytic myocarditis (n =1), and Lambl's excrescences (n = 4). To our best knowledge Lambl's excrescences, aneurysm of the pulmonary trunk, and cirsoid aneurysms have not previously been described in marine mammals, and some of these findings should be taken into account as possible causes of dolphin morbidity, mortality, and stranding.

Immunofluorescence staining for the detection of immunoglobulins and complement (C3) in dogs with renal disease.

Renal cortical biopsies from 74 dogs with different degrees of renal failure were studied by immunofluorescence to assess the frequency and extent of the deposition of immunoglobulins G, M and A (IgG, IgM, IgA) and complement C3. The dogs were divided into two groups on the basis of their clinical signs, and standard histological and electron microscopical examinations, according to whether their disease was an immune-mediated nephropathy (IMN) or a non-immune-mediated nephropathy (NIMN). In the dogs with an imn there was strong immunofluorescence due to IgG in the mesangium and the glomerular basement membrane and to IgM in the mesangium. The mechanism of immune complex trapping in the glomerulus also resulted in positive reactions to IgM in the dogs with an NIMN.

Thermal and viscoelastic properties of kappa/iota-hybrid carrageenan gels obtained from the Portuguese seaweed Mastocarpus stellatus.

The potential use of the underexploited Portuguese seaweed Mastocarpus stellatus as a source of natural thickening and gelling agents for food applications has been investigated. The alkaline pretreatment duration and extraction parameters (pH, temperature, and extraction duration) of seaweeds have been systematically varied in order to produce kappa/iota-hybrid carrageenans exhibiting a wide range of chemical properties. The mechanical spectra and the setting and melting temperatures of gels obtained by cooling 1.5 wt % extracted biopolymers solutions in 0.05 mol/dm3 KCl were measured by means of small amplitude oscillatory shear experiments. Gels showing elastic storage moduli ranging from 250 to 2750 Pa with no water syneresis could be obtained, and the relationships between the thermal properties and elasticity of the gels were evidenced. Gels mechanical properties are shown to correlate well with polysaccharides chemical parameters such as the degree of sulfate groups, the molecular weight distribution, and the relative content in iota-carrageenan monomers as determined by FTIR spectroscopy.

Effect of extraction parameters on the chemical structure and gel properties of kappa/iota-hybrid carrageenans obtained from Mastocarpus stellatus.

Extraction parameters (temperature, pH, duration) and alkaline pre-treatment duration have been systematically varied in the aim of exploring their impact on both chemical structure and gelling properties of carrageenan biopolymers obtained from Mastocarpus stellatus seaweeds, collected on the Northern coast of Portugal. Increasing the alkaline pre-treatment duration PT leads to kappa/iota-hybrid carrageenans containing less sulphate groups and biological precursor monomers. As a result, gel properties in the presence of KCl are improved as demonstrated by the increase in the Young's modulus with parameter PT. Increasing the extraction duration t ameliorates the biopolymer yield with no significant change in the complex kappa/iota-hybrid carrageenan chemical structure. However, smaller molecular weights are obtained and gel properties are seen to be negatively affected. Extraction temperature and pH have dramatic effects on the biopolymer gel strength, and a set of extraction parameters optimized with respect to extraction yield and gel properties is reported. In addition, kappa/iota-hybrid carrageenans obtained throughout this study exhibit a wide range of gel strengths in KCl, and allow us to present correlations between gel thermal properties and the kappa/iota-hybrid carrageenans chemical structure.

Long-term tolerability of tramadol LP, a new once-daily formulation, in patients with osteoarthritis or low back pain.

INTRODUCTION: Tramadol hydrochloride is a centrally acting analgesic, which possesses opioid agonist properties and activates monoaminergic spinal inhibition of pain. An oral, once a day, sustained release formulation of tramadol is thought to be advantageous compared with immediate release preparations as it prevents plasma peaks associated with increased side-effects of the drug. It may also improve compliance. The purpose of the study was to assess the long-term safety of a new sustained-release formulation of tramadol (tramadol LP) in patients with knee or hip osteoarthritis and in patients with refractory low back pain. STUDY DESIGN: The design was a phase III, open, multicentre, international, tolerability study with tramadol LP at a dose titrated by the patient between 100 and 400 mg once daily, according to the intensity of pain. The treatment was administered for a continuous period of 4 weeks followed by an intermittent intake of 5 months in 204 patients. The safety criteria for evaluation were recording of adverse events, laboratory tests, electrocardiogram, radiography, global tolerability assessed by the patient and the investigators. RESULTS: Long-term use of tramadol LP was reasonably well tolerated. Most of the reported adverse events were expected and occurred within the first month of treatment. Roughly half of the patients (49%) reported adverse events, of which 66% were related to treatment. Gastrointestinal events (nausea and vomiting) were the most frequent. Serious adverse events were reported in 6.4% of patients, from which only two cases were related to treatment. There was no sign of tolerance development and the percentage of patients presenting withdrawal symptoms after the end of treatment was low (6%). CONCLUSION: Long-term treatment with tramadol LP once daily is generally safe in patients with osteoarthritis or refractory low back pain.

Pharmacokinetic evaluation of a new oral sustained release dosage form of tramadol.

AIMS: To compare the pharmacokinetic profile of a new modified release formulation of tramadol (Tramadol LP 200 mg, SMB Technology, Marche-en-Famenne, Belgium) with that of an immediate release capsule (Topalgic) 50 mg, Grünenthal, Aachen, Germany) after single and multiple dosing and to assess the potential effect of food on its relative bioavailability. METHODS: The first study had an open, single-dose, three-treatment, three-period, six-sequence, randomised, crossover design with at least a five-day wash-out. The second study had an open, steady-state, two-treatment, two-period, two-sequence, randomised crossover design with at least a seven-day wash-out. Both studies contained 30 healthy subjects. Both enantiomers of tramadol and O-demethyl-tramadol (the only active metabolite of tramadol) were assayed in the plasma using an LC-MS/MS method. AUC infinity, AUCt, Cmax, Tmax, and T1/2 were estimated. Statistical analysis was performed using univariate anova, the Wilcoxon nonparametric method or Friedman's nonparametric anova where appropriate. RESULTS: Tramadol had a significantly lower Cmax and longer Tmax than the conventional formulation. Thus, the mean (+/- sd) Cmax of tramadol were 646 +/- 192 and 300 +/- 94 ng ml-1 for Topalgic 4 x 50mg and Tramadol LP 200 mg, respectively (95% confidence interval on the difference expressed as a percentage 42-51). AUC of tramadol from both formulations was comparable (similar AUC infinity and AUCt). Thus, the mean AUC infinity of (+/-)tramadol obtained after multiple dosing were 4611 +/- 1944 and 5105 +/- 2101 ngh ml-1 after Topalgic 4 x 50mg and Tramadol LP 200 mg, respectively (95%CI 102-123%). We also demonstrate that the pharmacokinetics of the drug are not influenced by the intake of food. Thus, the mean AUC infinity of (+/-) tramadol were 5444 +/- 1637 and 5169 +/- 1580 ngh ml-1 after Tramadol LP 200 mg given in the fasting and fed states, respectively (95%CI = 88-103%). CONCLUSIONS: The new sustained release form of tramadol exhibits adequate properties for once a day administration. Furthermore, its pharmacokinetic profile is not affected by the intake of food.

Automatic orienting of visuospatial attention in Parkinson's disease.

The basal ganglia are involved in not only motor behavior, but also other more cognitive processes, such as attention. We tested Parkinson's disease (PD) patients in a task that measures reflexive orienting of spatial attention. Seven patients with idiopathic PD and eight control subjects performed a covert orienting task where spatial attention was directed by means of exogenous cues (luminance increments) with no predictive validity for target position. The subjects' task was to make a speeded saccade to a visual target, which appeared a variable time after onset of the cue either in the cued or an uncued spatial position. There was no overall difference between PD patients and control subjects in terms of the initial facilitation following reflexive cues, and later inhibition of return (IOR). However, PD patients differed from control subjects in two important respects. First, they were significantly faster than were control subjects on this reflexive visual-orienting task. Second, disease severity correlated with attentional performance; more advanced patients showed less initial facilitation but greater IOR. Thus PD patients show better performance on a reflexive saccade task and, for more advanced patients, greater IOR than control subjects. These findings are consistent with the possibility that reflexive attentional processes in PD patients may be more active.

Schizotypal traits, attention and eye movements.

1. Subjects demonstrating high, average, or low schizotypal traits participated in saccade tasks of eye movements and attention including: a simple saccade task, an antisaccade task, and/or a cued saccade task measuring both facilitatory effects of cuing and inhibition of return (IOR). 2. Subjects were recruited based on their scores on the Rust Inventory of Schizotypal Cognitions (RISC) and then were given Raine's Schizotypal Personality Questionnaire (SPQ) (1991). 3. Subjects scoring high in schizotypy demonstrated increased errors on the voluntary eye movement task (antisaccade task) (p<0.05). Performance on the reflexive saccade task was not impaired in high compared to low schizotypals, but may have been enhanced as demonstrated by a negative correlation between scores on the SPQ and performance on this task. For the cued saccade task, there were no overall differences in cueing effects between schizotypal groups, however there was a laterality difference between low versus high scoring schizotypal subjects. 4. These results indicate distinct differences in tasks of overt orienting (saccade and antisaccade tasks) and covert orienting tasks (cued saccade task). The patterns of performance by our schizotypy subjects, including impaired voluntary saccade, enhanced reflexive saccade, and lateralized performance on the cued saccade task, are consistent with the performance of schizophrenic patient populations. Thus, our study supports the previous findings of a physiological relationship between schizotypal personality and schizophrenia.

Inhibition of return in manual and saccadic response systems.

When nonpredictive exogenous visual cues are used to reflexively orient covert visual spatial attention, the initial early facilitation for detecting stimuli at cued versus uncued spatial locations develops into inhibition by 300 msec following the cue, a pattern referred to as inhibition of return (IOR). Experiments were carried out comparing the magnitude and time course for development of IOR effects when manual versus saccadic responses were required. The results showed that both manual and saccadic responses result in equivalent amounts of facilitation following initial exposure to a spatial cue. However, IOR developed more quickly for saccadic responses, such that, at certain cue-target SOAs, saccadic responses to targets were inhibited, whereas manual responses were still facilitated. The findings are interpreted in terms of a premotor theory of visual attention.

Control of voluntary and reflexive saccades in Parkinson's disease.

Eight patients with idiopathic Parkinson's disease (PD) were compared with a group of age-matched controls on both reflexive saccade and antisaccade tasks. While reflexive, visually guided saccades led to equivalent performance in both groups, PD patients were slower, made more errors, and showed reduced gain on antisaccades (AS). This is consistent with previous results showing that PD patients have no difficulty with reflexive saccades but show deficiencies in a number of voluntary saccade paradigms. Moreover, visual information in the form of landmarks improves AS performance more for PD patients than controls, a finding analogous to results seen with other motor acts such as target-directed pointing. Results are discussed in terms of a two-process model of attention and eye movements.

Shape selectivity in primate lateral intraparietal cortex.

The extrastriate visual cortex can be divided into functionally distinct temporal and parietal regions, which have been implicated in feature-related ('what') and spatial ('where') vision, respectively. Neuropsychological studies of patients with damage to either the temporal or the parietal regions provide support for this functional distinction. Given the prevailing modular theoretical framework and the fact that prefrontal cortex receives inputs from both temporal and parietal streams, recent studies have focused on the role of prefrontal cortex in understanding where and how information about object identity is integrated with (or remains segregated from) information about object location. Here we show that many neurons in primate posterior parietal cortex (the 'where' pathway) show sensory shape selectivities to simple, two-dimensional geometric shapes while the animal performs a simple fixation task. In a delayed match-to-sample paradigm, many neuronal units also show significant differences in delay-period activity, and these differences depend on the shape of the sample. These results indicate that units in posterior parietal cortex contribute to attending to and remembering shape features in a way that is independent of eye movements, reaching, or object manipulation. These units show shape selectivity equivalent to any shown in the ventral pathway.

Spatial selective attention in schizophrenic, affective disorder, and normal subjects.

Schizophrenic, affective disorder, and normal subjects performed tasks involving exogenous (automatic) and endogenous (voluntary) attention. In the exogenous attention task, schizophrenic subjects demonstrated a greater benefit in response time than did normal subjects. In the endogenous attention task, however, schizophrenic subjects showed a smaller benefit in response time than did normal subjects. These results are consistent with a model of schizophrenia that predicts a deficit in voluntary (endogenous) control, and a disinhibition and therefore enhancement of the automatic (exogenous) processes of spatial selective attention. Affective disorder subjects did not demonstrate a greater benefit in response time than normal subjects in the exogenous attention task, but did show a smaller benefit in response time than normal subjects in the endogenous attention task. The somewhat similar pattern of behavior of schizophrenic and affective disorder subjects suggests that abnormal spatial selective attentional processes may not be specific to schizophrenia.

Glu-96 of basic fibroblast growth factor is essential for high affinity receptor binding. Identification by structure-based site-directed mutagenesis.

The importance of basic fibroblast growth factor (bFGF) in several pathophysiological processes has stimulated interest in the design of receptor antagonists to mitigate such effects. Of key importance in this connection is the characterization of the functional binding epitopes of the growth factor for its receptor. Based on peptide mapping and molecular dynamics calculations of the three-dimensional structure of basic fibroblast growth factor, we employed site-directed mutagenesis to investigate the effect of altering residues at positions 107, 109-114, and 96 on bFGF on receptor binding affinity. All muteins were cloned and expressed in Escherichia coli, purified to homogeneity employing heparin-Sepharose columns, and evaluated for receptor binding affinity. We found that replacement of residues at positions 107 and 109-114 by alanine or phenylalanine had little effect on receptor binding affinities compared with wild type bFGF, in agreement with previous evidence that bFGF residues 109-114 comprise a low affinity binding site. By contrast, substitution of Glu-96 with alanine yielded a molecule having about 0.1% of the affinity of the wild type bFGF. The affinity of the corresponding lysine and glutamine muteins was 0.3 and 10%, respectively, emphasizing the importance of a negative charge at this position. Our findings are consistent with the view that residues 106-115 on bFGF represent a low affinity binding site on bFGF. In addition, we identify Glu-96 as a crucial residue for binding to fibroblast growth factor receptor-1.

Antisaccades and smooth pursuit eye movements in schizophrenia.

Saccadic and smooth pursuit eye movements were recorded in schizophrenic patients, nonschizophrenic psychiatric patients, and normal controls. Both schizophrenic subjects and psychiatric controls demonstrated greater increases in error rates and greater delays in generating antisaccades than did normal controls. Schizophrenic patients with impaired smooth pursuit tracking showed greater increases in error rates in the antisaccade task than did schizophrenic patients with normal pursuit. Among psychiatric controls, increased errors on the antisaccade task were unrelated to pursuit performance. The small size of this group, however, reduces the power to detect a relation between smooth pursuit tracking and performance on the antisaccade task. Although most patients were receiving one or more medications, some of which can affect eye movements, medication state in this study did not account for differences we report in dependent variables.

Express saccades and smooth pursuit eye movement function in schizophrenic, affective disorder, and normal subjects.

Abstract Saccadic and smooth pursuit eye movements were recorded in three groups of subjects: a schizophrenic group, a non-schizophrenic psychotic patient comparison group, and a normal control group. Schizophrenic subjects demonstrated a greater decrease in saccadic response time than did normal controls in a gap task (when the fixation point was turned off 150 msec before the target appeared). The psychiatric comparison subjects did not differ from normal controls. Further, only schizophrenic subjects demonstrated a relation between smooth pursuit and saccadic eye movement performance, such that subjects with impaired smooth pursuit showed a larger decrease in saccadic response time in the gap task. The relation between performance on the gap task and quality of smooth pursuit and its relevance for a prefrontal deficit hypothesis of schizophrenia are discussed.

A simplified surveillance case definition of AIDS derived from empirical clinical data. The Clinical AIDS Study Group, and the Working Group on AIDS case definition.

A clinical AIDS case definition is needed for surveillance in countries where the CDC case definition is not practical. To derive such a definition, we compared 110 HIV-seropositive and 135 randomly selected HIV-seronegative adult medical-ward inpatients in Brazil. Multivariate analysis of clinical signs and symptoms and simple diagnoses resulted in a discriminant function with sensitivity of 89% and specificity of 96% in predicting for AIDS. These data were the empirical basis for a clinical definition of AIDS in adults drafted in a Caracas, Venezuela, workshop sponsored by the Pan American Health Organization. The revised "Caracas" definition presented here requires a positive HIV serology, the absence of cancer or other cause of immunosuppression, plus > or = 10 cumulative points, as follows: Kaposi's sarcoma (10 points); extrapulmonary/noncavitary pulmonary tuberculosis (10); oral candidiasis or hairy leukoplakia (5); cavitary pulmonary/unspecified tuberculosis (5); herpes zoster < 60 years of age (5); CNS dysfunction (5); diarrhea > or = 1 month (2); fever > or = 1 month (2); cachexia or > 10% weight loss (2); asthenia > or = 1 month (2); persistent dermatitis (2); anemia, lymphopenia, or thrombocytopenia (2); persistent cough or any pneumonia except TB (2); and lymphadenopathy > or = 1 cm at > or = 2 noninguinal sites for > or = 1 month (2). This definition has a sensitivity of 95% and a specificity of 100% (91% without HIV serology) when applied to the Brazilian patients in this study. The Caracas definition has been adopted by Brazil, Honduras, and Surinam, and is in validation elsewhere. The use of a reasonably sensitive and specific case definition commensurate with available diagnostic resources should facilitate AIDS surveillance in developing countries.