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Enhancement of cellular and humoral immunity in young broilers by the dietary supplementation of beta-hydroxy-beta-methylbutyrate.

As a dietary supplement, beta-Hydroxy-beta-Methylbutyrate (HMB), a catabolite of leucine, has been shown to reduce broiler mortality. In a series of experiments, male broilers (Experiments 1 and 2, n = 576) were grown for 21 days on diets that contained HMB at 0, 0.01. 0.05, and 0.10% of diet. In Experiment 3 (n = 240), chicks were fed diets containing 0, 0.05, 0.075, and 0.10% HMB. HMB dietary supplementation did not significantly affect broiler weight gain in any experiment. However, a trend toward increased mean broiler weight gain per bird was observed in Experiments 1 and 3 when HMB was consumed at 0.10% of the diet. Mean feed to gain ratio was not affected by the inclusion of HMB in broiler diets. In Experiment 3, HMB supplemented diets did not affect bursa of Fabricius, thymus, and spleen weights at 21 days of age. Cutaneous basophilic hypersensitivity response against pokeweek mitogen was higher (P < or = 0.05) at 48 and 72 hours post-injection in chicks on 0.05% dietary HMB (Experiment 1). In Experiment 2, this increase occurred 24 hours post-injection in chicks fed HMB at 0.01% of the diet. On the contrary, the T-cell mediated response against PHA-P mitogen was comparable between all dietary treatments in multiple experiments. Macrophage function profiles were determined at 21 days of age. All chicks in experiments 1 and 2 on HMB supplemented diets showed an increase in the recruitment of Sephadex-G50-elicited abdominal exudate cells (AEC). A 2-fold increase in AEC numbers occurred at the 0.10% HMB level (Experiment 1, P < or = 0.05). Although HMB supplementation did not significantly affect the phagocytic potential of the abdominal macrophages, nitrite levels in the macrophage culture supernatants were higher in 0.01% and 0.05% treatment groups as compared to the controls (Experiment 2, P < or = 0.04; Experiment 3, P < or = 0.05). HMB supplementation did not alter the bird's ability to clear Escherichiacoli or Salmonella arizona from the bloodstream. Beginning 7 days post-hatch, chicks were injected i.v. with a 7% sheep red blood cells suspension. Serum samples were collected to determine the primary and secondary antibody response. Chicks receiving the 0.1% HMB diet in Experiments 1 and 2 exhibited increased IgG and total anti-sheep red blood cell (SRBC) antibody levels during the primary response. During the secondary response, birds consuming the 0.10% HMB diet had elevated IgM levels as well as increased total anti-SRBC levels over the controls in Experiments 1 and 3. These studies show that HMB supplementation improves several immunological functions in young broilers, and such improvement may result in decreased mortality.

Adjuvants, Immunologic↗

S18616, a highly potent, spiroimidazoline agonist at alpha(2)-adrenoceptors: I. Receptor profile, antinociceptive and hypothermic actions in comparison with dexmedetomidine and clonidine.

S18616 ((S)-spiro[(1-oxa-2-amino-3-azacyclopent-2-ene)-4, 2'-(8'-chloro-1',2',3',4'-tetrahydronaphthalene)]) displayed high affinity at native rat alpha(2)-adrenoceptors (AR)s (pK(i), 9.8), native human (h)alpha(2A)-ARs (9.6), and cloned halpha(2A)- (9.5), halpha(2B)- (9.2), and halpha(2C)- (9.0) ARs. It showed 40-fold lower affinity for halpha(1A)-ARs (8.4) and >/=100-fold lower affinity for rat alpha(1)-ARs (7.1), halpha(1B)-ARs (7.7), halpha(1D)-ARs (7.6), imidazoline(1) (7.4), and imidazoline(2) (7.4) sites and >100-fold lower affinity for all other (>50) sites. At halpha(2A)-ARs, in guanosine-5'-O-(3-[(35)S]thio)triphosphate binding studies, S18616 was a potent (partial) agonist: log effective concentration (pEC(50)), 9.3/maximal effect, 51%. This observation was corroborated employing a halpha(2A)-Gi1alpha fusion protein/GTPase assay (9.0/40%) in which the actions of S18616 were blocked by pertussis toxin. Employing guanosine-5'-O-(3-[(35)S]thio)triphosphate binding assays, S18616 was also a partial agonist at halpha(2C)-ARs (8.2/63%) but a full agonist (8.4/124%) at halpha(2B)-ARs. At halpha(2A)-, halpha(2B)-, and halpha(2C)-ARs, the selective alpha(2)-AR antagonist, atipamezole, abolished the actions of S18616: pK(b) values of 9.1, 9. 1, and 9.4, respectively. As determined by depletion of membrane-bound [(3)H]phosphatidyl inositols, S18616 behaved as a (less potent) agonist (7.8/79%) at halpha(1A)-ARs, an action abolished by prazosin (pK(b), 8.9). Reflecting alpha(2)-AR agonist properties, S18616 potently (>/=1 microg/kg, s.c.) and dose dependently elicited hypothermia and antinociception (nine diverse models) in rodents. These actions were dose dependently inhibited by chemically diverse alpha(2)- versus alpha(1)-AR antagonists, atipamezole, idazoxan, RX821,002, and BRL44418 (a preferential alpha(2A)-AR ligand). In contrast, the actions of S18616 were unaffected by the alpha(1)-AR antagonists, ARC239 and prazosin (which preferentially block alpha(2B/2C)- versus alpha(2A)-ARs). Although the affinity of dexmedetomidine at alpha(2)-ARs was lower than S18616; it displayed a similar receptor and functional profile. Clonidine displayed lower efficacy than S18616, was substantially less potent, and had marked affinity for imidazoline(1) sites and alpha(1)-ARs. In conclusion, S18616 is a novel, selective, and highly potent agonist at alpha(2)-ARs.

Adrenergic alpha-2 Receptor Agonists↗

H+,K+-ATPase.

The H+,K+-ATPases comprise a group of integral membrane proteins that belong to the X+,K+-ATPase subfamily of P-type cation-transporting ATPases. Although these H+,K+-ATPase isoforms share approximately 60-70% amino acid identity, they exhibit discrete kinetic and pharmacological properties when expressed in heterologous systems. HK alpha2 has been categorized by its insensitivity to Sch-28080, an inhibitor of the gastric H+,K+-ATPase, and partial sensitivity to ouabain, an inhibitor of the Na+,K+-ATPase. This functional profile contrasts with the pharmacological sensitivities ascribed to HK alpha2 in transport studies in rat isolated medullary collecting ducts perfused in vitro and in mouse medullary collecting duct cell lines. HK alpha2 mRNA and protein abundance appears to be both tissue and site-specifically upregulated in response to chronic hypokalemia. This regulatory response has been localized to the outer and inner medulla. To reconcile these expressed sensitivities to those reported in vitro in isolated tubules and cells in culture, it would be necessary to invoke modification of the pharmacologic insensitivity of the colonic H+,K+-ATPase to Sch-28080. Although a 'unique' beta-subunit has been reported recently, this beta-subunit (beta(c)) is identical at the amino acid level to the recently cloned beta3-Na+,K+-ATPase. Moreover, while HK alpha2 can assemble indiscriminately with any X+,K+-ATPase beta-subunit, HK alpha2 has been reported to assemble stably with beta1-Na+,K+-ATPase in the renal medulla and in the distal colon. It remains conceivable that subunit assembly could be tissue specific and might respond to different physiological and pathophysiological stimuli. Furthermore, recent studies have suggested that the H+,K+-ATPase is both Na+-dependent and localized to the apical membrane in the distal colon. Therefore, future studies will need to resolve these discrepancies by determining if a unique, yet undiscovered H+,K+-ATPase isoform exists in kidney, or if post-translational modifications of the alpha- and/or beta-subunits could account for these functional diversities.

Animals↗

[Work capacity and re-entry into the workforce for the cardiac patient].

BACKGROUND: Treatment and rehabilitation of patients with heart disease is focused on survival, work resumption, achievement of a state of subjective well-being and compliance with behavioural changes in order to prevent progression of the atherosclerotic process. OBJECTIVES: To propose a specific job fitness evaluation method for workers with heart disease, based on analysis of occupational factors influencing the cardiovascular system and the definition of a clinical and functional profile of the patient, with special regard to aspects influencing vocational attitude. METHODS: A literature review was performed. RESULTS AND CONCLUSION: Although resumption of work is not the sole objective in the rehabilitation process, it remains an important goal not only for economic reasons, but also because it is beneficial for most patients' psychological well-being. However, it must be admitted that specific job fitness evaluation is rarely performed during rehabilitation programmes of cardiac patients and this can cause problems at the time of return to work. Doubts and uncertainties are also due to the lack of standardized evaluation methods and to the clinical peculiarity of these patients, mainly due to the risk of sudden disability. The job fitness evaluation method used in this study allows physicians to perform assessments based on objective data, so improving patients' confidence in their work capacity. To achieve a satisfactory application of the method, close cooperation between the cardiologist and the occupational physician is necessary.

Disease Susceptibility↗

Differential actions of antiparkinson agents at multiple classes of monoaminergic receptor. II. Agonist and antagonist properties at subtypes of dopamine D(2)-like receptor and alpha(1)/alpha(2)-adrenoceptor.

The accompanying multivariate analysis of the binding profiles of antiparkinson agents revealed contrasting patterns of affinities at diverse classes of monoaminergic receptor. Herein, we characterized efficacies at human (h)D(2SHORT(S)), hD(2LONG(L)), hD(3), and hD(4.4) receptors and at halpha(2A)-, halpha(2B)-, halpha(2C)-, and halpha(1A)-adrenoceptors (ARs). As determined by guanosine 5'-O-(3-[(35)S]thio)triphosphate ([(35)S]GTPgammaS) binding, no ligand displayed "full" efficacy relative to dopamine (100%) at all "D(2)-like" sites. However, at hD(2S) receptors quinpirole, pramipexole, ropinirole, quinerolane, pergolide, and cabergoline were as efficacious as dopamine (E(max)100%); TL99, talipexole, and apomorphine were highly efficacious (79-92%); piribedil, lisuride, bromocriptine, and terguride showed intermediate efficacy (40-55%); and roxindole displayed low efficacy (11%). For all drugs, efficacies were lower at hD(2L) receptors, with terguride and roxindole acting as antagonists. At hD(3) receptors, efficacies ranged from 33% (roxindole) to 94% (TL99), whereas, for hD(4) receptors, highest efficacies (approximately 70%) were seen for quinerolane, quinpirole, and TL99, whereas piribedil and terguride behaved as antagonists and bromocriptine was inactive. Although efficacies at hD(2S) versus hD(2L) sites were highly correlated (r = 0.79), they correlated only modestly to hD(3)/hD(4) sites (r = 0.44-0.59). In [(35)S]GTPgammaS studies of halpha(2A)-ARs, TL99 (108%), pramipexole (52%), talipexole (51%), pergolide (31%), apomorphine (16%), and quinerolane (11%) were agonists and ropinirole and roxindole were inactive, whereas piribedil and other agents were antagonists. Similar findings were obtained at halpha(2B)- and halpha(2C)-ARs. Using [(3)H]phosphatidylinositol depletion, roxindole, bromocriptine, lisuride, and terguride displayed potent antagonist properties at halpha(1A)-ARs. In conclusion, antiparkinson agents display diverse agonist and antagonist properties at multiple subtypes of D(2)-like receptor and alpha(1)/alpha(2)-AR, actions, which likely contribute to their contrasting functional profiles.

Adrenergic alpha-1 Receptor Agonists↗

Low-T3 syndrome: a strong prognostic predictor of death in patients with heart disease.

BACKGROUND: Clinical and experimental data have suggested a potential negative impact of low-T3 state on the prognosis of cardiac diseases. The aim of the present prospective study was to assess the role of thyroid hormones in the prognosis of patient population with heart disease. METHODS AND RESULTS: A total of 573 consecutive cardiac patients underwent thyroid function profile evaluation. They were divided in two subgroups: group I, 173 patients with low T3, ie, with free T3 (fT3) <3.1 pmol/L, and group II, 400 patients with normal fT3 (>or=3.1 pmol/L). We considered cumulative and cardiac death events. During the 1-year follow-up, there were 25 cumulative deaths in group I and 12 in group II (14.4% versus 3%, P<0.0001); cardiac deaths were 13 in group I and 6 in group II (7.5% versus 1.5%, P=0.0006). According to the Cox model, fT3 was the most important predictor of cumulative death (hazard ratio [HR] 3.582, P<0.0001), followed by dyslipidemia (HR 2.955, P=0.023), age (HR 1.051, P<0.005), and left ventricular ejection fraction (HR 1.037, P=0.006). At the logistic multivariate analysis, fT3 was the highest independent predictor of death (HR 0.395, P=0.003). A prevalence of low fT3 levels was found in patients with NYHA class III-IV illness compared with patients with NYHA class I-II (chi(2) 5.65, P=0.019). CONCLUSIONS: Low-T3 syndrome is a strong predictor of death in cardiac patients and might be directly implicated in the poor prognosis of cardiac patients.

Aged↗

A cytochemical and immunohistochemical approach to malignant histiocytosis.

Malignant histiocytosis (MH) is a true histiocytic disorder, whose identification is still based on too broad morphologic criteria. Using routine histology, cytochemical and immunohistochemical techniques on involved lymph nodes, 15 cases of MH have been investigated. Pleomorphism and cellular atypia, phagocytosis, lack of cohesiveness between proliferating cells, sinusoidal involvement, and plasmacytic infiltrate were the most common histologic features. MGG-stained imprints from 14 cases showed a composite tumor population mainly consisting of histiocyte-appearing cells, poorly differentiated atypical cells, and multinucleated giant cells. These cells, irrespective of cytologic features, revealed a diffuse, moderately to strongly positive reaction with acid phosphatase and nonspecific esterase. Naphthol-AS-D-chloroacetate esterase, Sudan black B, alkaline phosphatase, and beta-glucuronidase reactions were completely negative. Immunoperoxidase studies in 11 cases demonstrated that tumor cells stained positively for both kappa and lambda chains. These cells were also positive for albumin. Polytypic staining for IgG was observed in two cases, and a weak staining for lysozyme was found in two other nodes. Global results confirm the value of these studies for functional profile determination of MH proliferating cells. A combined approach using a variety of cytochemical and immunohistochemical techniques should be routinely considered in MH as useful additional studies for a more precise diagnostic definition of the disease.

Adolescent↗

A study of the influence of polysaccharides on collagen self-assembly: nanostructure and kinetics.

Collagen, a critical part of the extra-cellular matrix of tissues, is a popular native material for building scaffolding for tissue-engineering applications. To mimic the structural and functional profiles of materials found in the native extra-cellular matrix, numerous efforts have been made toward developing a novel scaffold combining collagen with other biomacromolecules. All of these works have been focused on improving the mechanical or biochemical properties of the collagen-based matrix. Unfortunately, most of these studies have failed to consider the nanostructure of collagen in the complex matrix. The aim of our study was to investigate the aggregation pattern of collagen after addition of polysaccharides with positive or negative charge, the dose-response relationship, and the effect on reconstitution kinetics. Generally, collagen self-assembles into fibrils with a diameter of around 95 nm but, in the presence of various polysaccharides in varying amounts, collagen self-assembles into different shapes with larger diameters compared with collagen alone. Although the morphology and diameter of the collagen fibrils varies with reconstitution conditions, the D-periods of the fibrils all remained the same regardless of the species or concentration of polysaccharides. The kinetics of fibril formation was determined from turbidity-time curves. All turbidity curves demonstrated that polysaccharides only alter the lag time and time frame of reconstitution, but have no significant effect on the mechanism of reconstitution. Together our data indicate that the presence of biomacromolecules can alter the kinetics and the 3D fibril ultrastructure of assembled collagen and that the consequent structural changes may affect cellular responses in medical applications.

Alginates↗

Paget's disease of the anus: a clinicopathological study.

Paget's disease of the anus is a rare disorder of controversial origin and is frequently associated with malignancy. We studied eight patients and carried out immunohistochemical studies to determine whether particular functional profiles might be indicators of a malignant association. One patient presented with synchronous carcinoma and two developed carcinomas 3 and 10 years after excision of Paget's disease. Five patients underwent wide local excision and have not developed cancer (median follow-up 6 years, range 5-13 years). However, four patients developed recurrent Paget's disease. Immunohistochemical studies showed that in general Paget cells stained positively with CAM 5.2 (a cytokeratin marker), gross cystic disease fluid protein (a marker for apocrine cells), human milk fat globule glycoprotein (HMFG 1 and 2) and carcinoembryonic antigen but negatively for PR3A5 (a marker for colonic goblet cells). Three cases had a staining profile which was quite different from that usually observed and these were associated with malignancy. One showed an antigenic profile more typical of a large bowel carcinoma. Paget's disease of the anus appears to run one of two clinical courses: to develop malignancy; or to recur locally, often on repeated occasions. Wide local excision is the treatment of choice but long-term follow-up is necessary because of the cancer risk. An immunohistochemical staining pattern which is different from usual may indicate a higher malignant risk and/or identify some cases of Paget's disease as representing a downward 'pagetoid' extension from a anorectal adenocarcinoma rather than a true epidermotropic apocrine neoplasm of the perianal skin.

Aged↗

Heterogeneous expression of SNAP-25 and synaptic vesicle proteins by central and peripheral inputs to sympathetic neurons.

Neurons in prevertebral sympathetic ganglia receive convergent synaptic inputs from peripheral enteric neurons in addition to inputs from spinal preganglionic neurons. Although all inputs are functionally cholinergic, inputs from these two sources have distinctive neurochemical and functional profiles. We used multiple-labeling immunofluorescence, quantitative confocal microscopy, ultrastructural immunocytochemistry, and intracellular electrophysiologic recordings to examine whether populations of inputs to the guinea pig coeliac ganglion express different levels of synaptic proteins that could influence synaptic strength. Boutons of enteric intestinofugal inputs, identified by immunoreactivity to vasoactive intestinal peptide, showed considerable heterogeneity in their immunoreactivity to synaptosome-associated protein of 25 kDa (SNAP-25), synapsin, synaptophysin, choline acetyltransferase, and vesicular acetylcholine transporter. Mean levels of immunoreactivity to these proteins were significantly lower in terminals of intestinofugal inputs compared with terminals of spinal preganglionic inputs. Nevertheless, many boutons with undetectable levels of SNAP-25 immunoreactivity formed morphologically normal synapses with target neurons. Treatment with botulinum neurotoxin type A (20-50 nM for 2 hours in vitro) generated significant cleavage of SNAP-25 and produced similar dose- and time-dependent inhibitions of synaptic transmission from all classes of inputs, regardless of their mean level of SNAP-25 expression. The simplest interpretation of these results is that only synaptic boutons with detectable levels of SNAP-25 immunoreactivity contribute significantly to fast cholinergic transmission. Consequently, the low synaptic strength of intestinofugal inputs to final motor neurons in sympathetic pathways may be due in part to the low proportion of their boutons that express SNAP-25 and other synaptic proteins.

Adrenergic Fibers↗

Residual effects of zaleplon and zolpidem following middle of the night administration five hours to one hour before awakening.

The objective was to assess residual effects of zaleplon and zolpidem after a middle of the night administration. This was a randomized, double-blind, placebo-controlled, crossover study, conducted in 40 healthy young male and female subjects. Subjects were awakened in the middle of the night and administered either placebo or zaleplon 10 or 20 mg or zolpidem 10 mg. A battery of objective tests exploring psychomotor and cognitive functions such as critical flicker fusion (CFF), choice reáction time (CRT), digit symbol substitution test (DSST), and memory tests (Sternberg memory scanning and a word list) were administered immediately after morning waking. Zaleplon 10 mg was devoid of residual effects whatever - the time of dosing - except a minimal but significant decrease in DSST scores when administered 1 h before awakening. Zaleplon 20 mg produced significant residual effects on performance (increase in CRT, and decrease in CFF threshold and in DSST scores) and memory (decrease in immediate and delayed free recall of words) only when administered 1 h before awakening. In contrast, zolpidem 10 mg produced significant detrimental residual effects on CRT and delayed free recall of words, when administered up to 5 h before waking, on DSST and Sternberg when administered up to 3 h before awakening and on CFF when administered 1 h before awakening. The residual effects of zolpidem 10 mg were more marked than those observed after zaleplon 20 mg. The present results demonstrate that zaleplon 10 mg has no or minimal residual effects when administered in the middle of the night as little as 1 h before waking. The lack of clinically significant residual effects with zaleplon may be explained by its unique pharmacokinetic (rapid elimination half-life) and pharmacodynamic (selective binding for GABA(A) receptors with the alpha(1) subunit, dissociation between sleep inducing properties and impairment of cognitive functions) profiles. Copyright 2001 John Wiley & Sons, Ltd.

Journal Article↗

Metagenome-Based Characterization of the Gut Virome Signatures in Patients With Gout.

The gut microbiome has been implicated in the development of autoimmune diseases, including gout. However, the role of the gut virome in gout pathogenesis remains underexplored. We employed a reference-dependent virome approach to analyze fecal metagenomic data from 102 gout patients (77 in the discovery cohort and 25 in the validation cohort) and 86 healthy controls (HCs) (63 and 23 in each cohort). A subset of gout patients in the discovery cohort provided longitudinal samples at Weeks 2, 4, and 24. Our analysis revealed significant alterations in the gut virome of gout patients, including reduced viral richness and shifts in viral family composition. Notably, Siphoviridae, Myoviridae, and Podoviridae were depleted, while Quimbyviridae, Retroviridae, and Schitoviridae were enriched in gout patients. We identified 359 viral operational taxonomic units (vOTUs) associated with gout. Enriched vOTUs in gout patients predominantly consisted of Fusobacteriaceae, Bacteroidaceae, and Selenomonadaceae phages, while control-enriched vOTUs included Ruminococcaceae, Oscillospiraceae, and Enterobacteriaceae phages. Longitudinal analysis revealed that a substantial proportion of these virome signatures remained stable over 6 months. Functional profiling highlighted the enrichment of viral auxiliary metabolic genes, suggesting potential metabolic interactions between viruses and host bacteria. Notably, gut virome signatures effectively discriminated gout patients from HCs, with high classification performance in the validation cohort. This study provides the first comprehensive characterization of the gut virome in gout, revealing its potential role in disease pathogenesis and highlighting virome-based signatures as promising biomarkers for gout diagnosis and future therapeutic strategies.

Humans↗

A reference map of a human pituitary adenoma proteome.

In order to compare the proteomes from different cell types of pituitary adenomas for our long-term goal to clarify the molecular mechanisms that participate in the formation of pituitary adenoma, and to detect any tumor-related marker for an "early-stage" diagnosis, the two-dimensional gel electrophoresis (2-DE) reference map of a pituitary adenoma tissue proteome is described here. A vertical, two-dimensional (2-D) polyacrylamide gel electrophoresis system and PDQuest image analysis software have been used to provide a high level of between-gel reproducibility and to accurately array each protein expressed in a pituitary adenoma tissue. Mass spectrometry (matrix-assisted laser desorption/ionization-time of flight MALDI-TOF and liquid chromatography-electrospray ionization-quadrupole-ion trap LC-ESI-Q-IT) and protein databases were used to characterize each protein in the 2-D gel. The results demonstrate that a good reproducibility of the 2-D gel pattern was attained. The position deviation of matched spots among four 2-D gels was 1.95 +/- 0.45 mm in the isoelectric focusing direction, and 1.70 +/- 0.53 mm in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis direction. A total of ca. 1000 protein spots were separated by 2-DE, and 135 protein spots that represent 111 proteins were characterized with mass spectrometry (96 spots for MALDI-TOF, 39 spots for LC-ESI-Q-IT). The characterized proteins include pituitary hormones, cellular signals, enzymes, cellular-defense proteins, cell-structure proteins, transport proteins, etc. Those proteins were located in the cytoplasmic, cellular membrane, mitochondrial, endoplasmic reticulum, nuclear, ribonucleosome, extracellular fractions, or were secreted in plasma, etc. Those identified proteins contribute to a functional profile of the pituitary adenoma proteome. These data will be used to expand the proteome database of the human pituitary, which can be accessed in the website http://www.utmem.edu /proteomics.

Adenoma↗

The hepatoprotective effects of Limonium sinense against carbon tetrachloride and beta-D-galactosamine intoxication in rats.

In the present study, the hepatoprotective action of Limonium sinense (Plumbaginaceae) was evident after carbon tetrachloride (CCl(4)) and beta-D-galactosamine (D-GalN), respectively, challenge in rats. The plant materials were divided into two parts: (1) the roots extracted with water (WRE) and (2) the leaves extracted with methanol and fractionated with chloroform (CLE). Both WRE and CLE were extremely flavonoid-enriched extracts. In an CCl(4)-induced acute liver damage study, pretreatment with WRE at 300 mg/kg i.p. and CLE at 100 mg/kg i.p. significantly reduced the amino-transaminases levels of SGOT (p < 0.01) and SGPT (p < 0.01) previously increased by CCl(4) intoxication. In D-GalN-induced acute liver damage study, administration of WRE (300 and 500 mg/kg) or CLE (100 mg/kg) p.o. also significantly reduced the SGOT (p < 0.01) and SGPT (p < 0.01) levels previously increased by D-GalN intoxication. Furthermore, the serum triglyceride level was increased after pretreatment with WRE or CLE previously reduced by D-GalN intoxication. All of the liver function profiles were confirmed to have improvement of liver lesion in histopathological observation. In an acute toxicity test on ICR mice, the LD(50) of WRE was 777.6 mg/kg i.p. An in vitro study showed that CLE possessed a more potent cytotoxicity to human hepatocellular carcinoma cells (Hep3B) (EC(50) = 43.1 micro g/mL) than the other organic fractions, which were fractionated from methanol extracts of the leaves of L. sinense. The present results conclude that L. sinense possesses a hepatoprotective efficacy, and is relatively safe in rats.

Administration, Oral↗

Nested, non-parametric, correlative analysis of microarrays for heterogenous phenotype characterization.

We present a non-parametric approach for qualitatively selecting candidate genes to characterize several criteria that are nested among genes selected on the basis of their individual, similar effects upon an array-wide closeness measure. In this setting, a goal is to obtain a reliable characterization of phenotypes, based on very high-dimensional data from a few samples. As opposed to a distance-based approach, the proposed measure defines closeness based on gene signal profiles (functionals) rather than on isolated (numerical) differences in each gene between samples. By using such a measure to characterize intensity differences, we effectively separate biological from artifactual variation in expression, due to tissue effects or signal calibration. Based on this measure, we successively examine the significance of the following: a set of similarly behaved genes relative to all arrayed genes, a set of candidate genes relative to similarly behaved genes, individual candidate genes relative to non-candidates, and the direction, as over- or under-expressed, of candidate genes. In each setting, sample pairs are the units of analysis, with U-statistics the theoretical framework. We illustrate the method on a microarray experiment, where the goal is to select sets of genes that characterize a type of skin cancer and its histological subtypes.

Genetic Heterogeneity↗

Protease-activated receptor-2 (PAR-2): structure-function study of receptor activation by diverse peptides related to tethered-ligand epitopes.

Protease-activated receptor-2 (PAR-2) is a tethered-ligand, G-protein-coupled receptor that is activated by proteolytic cleavage or by small peptides derived from its cleaved N-terminal sequence, such as SLIGRL-NH2. To assess specific PAR activity, we developed an immortalized murine PAR-1 (-/-) cell line transfected with either human PAR-2 or PAR-1. A "directed" library of more than 100 PAR agonist peptide analogues was synthesized and evaluated for PAR-2 and PAR-1 activity to establish an in-depth structure-function profile for specific action on PAR-2. The most potent agonist peptides (EC50 = 2-4 microM) had Lys at position 6, Ala at position 4, and pFPhe at position 2; however, these also exhibited potent PAR-1 activity (EC50 = 0.05-0.35 microM). We identified SLIARK-NH2 and SL-Cha-ARL-NH2 as relatively potent, highly selective PAR-2 agonists with EC50 values of 4 microM. Position 1 did not tolerate basic, acidic, or large hydrophobic amino acids. N-Terminal capping by acetyl eliminated PAR-2 activity, although removal of the amino group reduced potency by just 4-fold. At position 2, substitution of Leu by Cha or Phe gave equivalent PAR-2 potency, but this modification also activated PAR-1, whereas Ala, Asp, Lys, or Gln abolished PAR-2 activity; at position 3, Ile and Cha were optimal, although various amino acids were tolerated; at position 4, Ala or Cha increased PAR-2 potency 2-fold, although Cha introduced PAR-1 activity; at position 5, Arg or Lys could be replaced successfully by large hydrophobic amino acids. These results with hexapeptide C-terminal amides that mimic the native PAR-2 ligand indicate structural modes for obtaining optimal PAR-2 activity, which could be useful for the design of PAR-2 antagonists.

Amino Acid Motifs↗

Effects of quinolones on granulocyte function in vitro.

Granulocytes (PMN) play an important role in antibacterial host defence. The modulation of PMN-function by different antibiotics has been shown previously. Therefore, we investigated the possible immuno-modulatory effects of four new quinolone antibiotics: ciprofloxacin, norfloxacin, pefloxacin, ofloxacin. Pure PMN-preparations were obtained from healthy volunteers and incubated for 30 min at 37 degrees C with the different quinolones at different concentrations (0.1-1-5-10 mg/l). A complete PMN-functional profile was assayed. At all concentrations tested no significant differences were seen for myeloperoxidase content, superoxide production and chemiluminescence. A significant stimulation of phagocytosis and killing occurred at 10 mg/l of norfloxacin and at 10 mg/l of pefloxacin. An increase of chemotaxis towards formyl-methionyl-leucyl-phenylalanine was noted at 10 mg/l of ofloxacin, while stimulation of hexose monophosphate shunt was seen at 1 mg/l of ciprofloxacin and 10 mg/l of ofloxacin. The mechanism whereby both subinhibitory and inhibitory concentrations of quinolones moderately influence PMN-responsiveness is not yet known. Further studies are needed to elucidate the nature of this quinolone-effect and to assess its relevance in vivo.

Anti-Infective Agents↗

Autoreactive eosinophils associated with spontaneous autocytotoxicity in bronchial asthma.

Allergic autocytotoxicity (ACT) assay is extensively investigated as an in vitro equivalent of oral ingestion challenge with food antigens among patients with food hypersensitivity. In direct ACT, food antigenic determinants interact directly with plasma cell membranes of sensitive individuals. Antibody-dependent ACT is also known as antibody-dependent-cell-mediated-cytotoxicity (ADCC) phenomenon, when food antigens combine with specific antibody through cell membranes Fc receptors among normal and sensitive individuals. Spontaneous ACT is a separate mechanism of white blood cells disintegration which does not require in vitro priming of the cells, neither by antigen or antibody. Spontaneous ACT occurs in some individuals as "background noise" in addition to direct and antibody-dependent ACT. The exact cellular nature of ACT phenomena are unknown at the present time with the exception that the common identifying factor for each of them is the disintegration and death of human white blood cells. Electron microscopy studies among four bronchial asthma patients with spontaneous ACT demonstrated eosinophils with atypical crystalloid cores and diffuse autolytic pattern of granular membranes. These ultrastructural characteristics are associated with new functional profiles of eosinophils expressed morphologically as natural killer and/or suicidal potency. At least two subpopulations of eosinophils are mediating ACT. The first subpopulation has normal ultrastructure observed in direct ACT and the second subpopulations has altered morphology of eosinophils granules described in spontaneous ACT. The natural killer-suicidal eosinophils presented in patient with spontaneous ACT illustrate a new pathway of cytodestructive mechanism in anaphylactic injury.

Adolescent↗