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PubMed1997

Allelic variations in the human growth hormone-1 gene promoter of growth hormone-deficient patients and normal controls.

OBJECTIVE: Isolated growth hormone deficiency (IGHD) type IB is suggested to be more probably due to alterations in the genes directly involved in the hypothalamo-pituitary axis and/or in the specific transcriptional regulation (cis-trans coupling) of the hGH-1 gene than to alterations in the gene itself. In this study we analyzed the hGH-1 gene promoter region for structural alterations and allelic variations. METHODS: The hGH-1 gene promoter region was analyzed by PCR, cycle sequencing and direct-blotting electrophoresis in a total of 212 individuals including 113 patients with IGHD type IB, 21 unaffected family members and 78 normal controls. RESULTS: Twenty-two sequence variation sites were identified. Of these, 14% were located around the region of -1075bp, 77% between -550bp and the translational start site (+1bp) and 9% within the first intron. Only one variation site affected a characterized cis-acting element, namely that of NF-1. Importantly, all the variations found in patients were also observed in non-affected family members as well as in normal unrelated controls. CONCLUSIONS: These findings imply that it is not a single variation within the GH-1 gene promoter, and therefore in the cis-acting elements, which causes IGHD. However, we can not exclude the possibility that combinations of variations might perturb expression. Furthermore, these data illustrate the normal heterogeneity of the GH-1 gene promoter region, a fact that has to be borne in mind whenever transcriptional studies are performed.

Alleles↗
PubMed1988

Effects of dietary clenbuterol on metabolism of the hindquarters in steers.

The objective of this study was to measure acute (d 1) and chronic (d 9) effects of dietary clenbuterol on heart rate, blood flow, oxygen uptake, and net uptake/release of metabolites in the hindquarters of growing steers. The design was a single reversal with two 9-d periods of control or 8 mg clenbuterol/d with 5 d between periods. Within 2 h of initial consumption of 2 mg clenbuterol (d 1), heart rate and blood flow doubled and arterial plasma concentrations of glucose, L-lactate and nonesterified fatty acid (NEFA) increased, whereas alpha-NH2 N and NH3 concentrations decreased, demonstrating an acute response. Uptake of oxygen increased and net uptake of alpha-NH2 N decreased. Net release of both L-lactate and NEFA increased. On d 9, there were no acute responses to clenbuterol consumption; however, heart rate, blood flow, and NEFA concentration remained chronically elevated, and plasma concentrations of acetate and propionate decreased compared with control feeding. Net uptake of alpha-NH2 N, oxygen and release of L-lactate by the hindquarters chronically increased on d 9 of clenbuterol feeding. Changes in both blood flow and arteriovenous (AV) concentration difference contributed to changes in uptake/release. The chronic metabolic changes and oxygen uptake were consistent with increased N retention in the hindquarters through increased protein synthesis, decreased use of acetate and increased reliance on NEFA for cellular energy. In conclusion, the data show that the perturbation of homeostatic regulation by dietary clenbuterol on d 1 evolved to establishment of homeorhetic regulation by d 9 that is consistent with increased skeletal protein accretion in growing steers.

Animals↗
PubMed1999

Seminal vesicle autoantigen, a novel phospholipid-binding protein secreted from luminal epithelium of mouse seminal vesicle, exhibits the ability to suppress mouse sperm motility.

Seminal vesicle autoantigen (SVA) is a 19 kDa glycoprotein purified from mouse seminal vesicle secretion. It was quantified to be 0.9% (w/v) in the seminal vesicle fluid. We examined its distribution in the accessory sexual gland, characterized its binding sites on the sperm surface and assessed its effect on sperm motility. It was immunolocalized on the epithelium of the primary and secondary folds in the tissue. Mouse spermatozoa collected from caudal epididymis were devoid of SVA. A cytochemical study illustrated the presence of SVA-binding region on the entire cells. The cytochemical staining intensity for the binding of SVA to spermatozoa remained even when the cells were pretreated with protease digestion, acid or heat at 100 degrees C for 10 min. Moreover, the SVA-sperm binding could be inhibited by the dispersed sperm lipid. The specificity of interaction between (125)I-SVA and phospholipids was studied by TLC overlay techniques. The radiolabelled protein showed strong binding to purified phosphatidylcholine and phosphatidylserine and weak binding to purified sphingomyelin, lysophosphatidylcholine and phosphatidylethanolamine, but did not interact with phosphatidic acid, lysophosphatidic acid or phosphatidylinositol. Among the lipids extracted from spermatozoa, SVA showed strong binding to phosphatidylcholine and weak binding to sphingomyelin and neutral lipids. The assay for SVA-sperm binding with (125)I-SVA determined the IC(50) as being (3.89+/-0.65)x10(-5) M(-1), which is compatible with an apparent dissociation constant of (9.10+/-0.02)x10(-5) M(-1) estimated by fitting the data of phosphatidylcholine-perturbed SVA fluorescence to a modified Scatchard plot. SVA showed an ability to suppress sperm motility. The average path velocity, straight-line velocity and curvilinear velocity of sperm were not detectable by computer-assisted sperm assay after incubation of the cells in the presence of 0.3% SVA at 37 degrees C for more than 40 min.

Animals↗
PubMed2000

Proliferative lesions and reproductive tract tumors in male descendants of mice exposed developmentally to diethylstilbestrol.

Prenatal exposure to diethylstilbestrol (DES) is associated with reproductive tract abnormalities, subfertility and neoplasia in experimental animals and humans. Studies using experimental animals suggest that the carcinogenic effects of DES may be transmitted to succeeding generations. To further evaluate this possibility and to determine if there is a sensitive window of exposure, outbred CD-1 mice were treated with DES during three developmental stages: group 1 was treated on days 9-16 of gestation (2.5, 5 or 10 microg/kg maternal body weight) during major organogenesis; group II was treated once on day 18 of gestation (1000 microg/kg maternal body weight) just prior to birth; and group III was treated on days 1-5 of neonatal life (0.002 microg/pup/day). DES-exposed female mice (F(1)) were raised to maturity and bred to control males to generate DES-lineage (F(2)) descendants. The F(2) males obtained from these matings are the subjects of this report; results in F(2) females have been reported previously [Newbold et al. (1998) CARCINOGENESIS:, 19, 1655-1663]. Reproductive performance of F(2) males when bred to control females was not different from control males. However, in DES F(2) males killed at 17-24 months, an increased incidence of proliferative lesions of the rete testis and tumors of the reproductive tract was observed. Since these increases were seen in all DES treatment groups, all exposure periods were considered susceptible to perturbation by DES. These data suggest that, while fertility of the DES F(2) mice appeared unaltered, increased susceptibility for tumors is transmitted from the DES 'grandmothers' to subsequent generations.

Animals↗
PubMed2001

[Eradication of Helicobacter pylori. Is it necessary to eradicate Helicobacter pylori in gastric reflux?].

POTENTIAL PROTECTIVE EFFECT OF H. PYLORI: Epidemiological studies have not demonstrated an association between H. pylori infection and symptoms of gastroesophageal reflux, reflux esophagitis, or Barrett's esophagus with or without dysplasia or esophageal adenocarcinoma. On the contrary, an apparently favorable negative association has been identified suggested a potential protective effect of H. pylori. An inverse association is also observed between the severity of reflux complications and infection by strains of H. pylori expressing certain virulence markers (cogA) associated with the more severe gastric lesions. The prevalence of gastroesophageal reflux-related disease has increased steadily for more than fifty years while the incidence of H. pylori infection has decreased in developed countries. This observation might suggest that H. pylori infection plays a protective role. In addition, eradication of H. pylori could favor he development of gastroesophageal reflux or reflux esophagitis. The degree of risk is unknown. Pangastritis with significant lesions of the gastric body leading to a reversible decrease in the secretion of acid after H. pylori eradication might lower the risk of gastroesophageal reflux. POTENTIAL PROMOTION EFFECT: The relations between H. pylori and gastroesophageal reflux are complex. Reflux associated with duodenal ulcer appears to be improved by eradication of H. pylori suggesting that infection might promote the development of reflux. The causal mechanism would be increased acid secretion induced by antral gastritis predominantly resulting from the H. pylori infection. Further work is needed to determine the exact role of gastritis and perturbed acid secretion. PERSPECTIVES: Data on the role of H. pylori in transient relaxation of the lower esophageal sphincter, implicated in the pathogenesis of gastroesophageal reflux, are lacking. Antisecretion treatments reduce the quantity of gastric acid favoring H. pylori colonization of the fundic mucosa and possibly aggravating fundic gastritis. The risk of progression from H. pylori gastritis to atrophy or intestinal metaplasia of the fundus under prolonged proton pump inhibitor treatment remains to be determined. Eradication of H. pylori reduces the efficacy of antisecretory drugs according to poorly understood mechanisms. The potential effect of gastroesophageal reflux on the transmission of H. pylori infection remains to be established. PRACTICAL ATTITUDE: Much remains to be learned concerning the beneficial or deleterious effects of eradication of H. pylori on the course of gastroesophageal reflux. In clinical practice, eradication of H. pylori is not useful for patients with gastroesophageal reflux.

Adenocarcinoma↗
PubMed1984

Relationships between the flux of lipid metabolism in vivo and the uncoupling of respiratory control.

The influence of dinitrophenol treatment on the balance of lipid metabolism in mouse tissues has been studied by measuring the incorporation in vivo of 3H- and 14C-labelled glycerol. Several significant alterations in the relative utilization of these labels were observed in treated animals, with the greatest influence being evident in liver lipids, and diverse responses occurring in kidney, heart, muscle and adipose tissue. These data establish that dinitrophenol perturbs lipid metabolism in all the major tissues of the animal, and point to the detailed nature of this response in the individual classes of neutral and phospholipids. The implications of individual changes have been discussed in relation to the relative roles of mitochondria and peroxisomes in the regulation of mammalian lipid metabolism.

Adipose Tissue↗
PubMed1995

Cyclic AMP mediates change in superoxide dismutase activity to monitor host-parasite interaction in Leishmania donovani.

The present study is designed to understand the role of cyclic AMP (cAMP) in host-parasite interaction involving Leishmania donovani, the causative agent for Kala-azar. When Leishmania promastigotes or macrophages were pretreated with dibutyryl cAMP or theophylline and epinephrine, which are well defined initiators for cAMP release, a key enzyme of the oxygen defense system, superoxide dismutase (SOD), was inhibited. At the same time, parasite interaction was considerably reduced to the level of 54.5% and 46.2%, respectively, for pretreated promastigotes. Internalization of the organisms in phagolysosomes was similarly affected. Dibutyryl cAMP-treated promastigotes in the presence of SOD, on the other hand, restored in vitro infection to the normal level. At least 50% less cAMP entered into Leishmania promastigotes when SOD was added to the incubation system containing dibutyryl cAMP. Data reveal that cAMP perturbs the Leishmania-macrophage interaction through inhibition of SOD, pointing to the importance of a promastigote enzyme for the survival of this pathogen within phagolysosomes.

Animals↗
PubMed1997

Pores formed by influenza hemagglutinin.

Low pH-induced fusion mediated by the hemagglutinin (HA) of influenza virus involves a conformational change in the protein that leads to the insertion of a "fusion peptide" of the protein into the target membrane. It has been suggested that this insertion, aided by the formation of a complex of multiple HA trimers, would lead to perturbation of the bilayer structure of the membrane, initiating fusion. Here we present data showing that the interaction of the bromelain released ectodomain of the protein (BHA) with liposomal membranes at low pH leads to pore formation, at least at low temperatures. Strongly temperature-dependent low pH-induced inactivation of BHA resulted in a complete lack of activity of BHA above 10 degrees C. Even at 0 degrees C, only about 5% of the BHA participated in pore formation. Viral HA was less rapidly inactivated and still induced pores at 37 degrees C. BHA-induced pore formation showed a sigmoidal time course. Once BHA had formed a pore in one liposome, it did not form a pore in a further liposome. Quantitative analysis of pore formation indicated that one single BHA trimer sufficed to produce a pore. These data indicate that fusion peptide insertion perturbs the membrane and that the formation of a complex of trimers is not a prerequisite for the perturbation.

Cold Temperature↗
PubMed2005

Solution of the inverse problem of magnetic induction tomography (MIT).

Magnetic induction tomography (MIT) of biological tissue is used to reconstruct the changes in the complex conductivity distribution inside an object under investigation. The measurement principle is based on determining the perturbation DeltaB of a primary alternating magnetic field B0, which is coupled from an array of excitation coils to the object under investigation. The corresponding voltages DeltaV and V0 induced in a receiver coil carry the information about the passive electrical properties (i.e. conductivity, permittivity and permeability). The reconstruction of the conductivity distribution requires the solution of a 3D inverse eddy current problem. As in EIT the inverse problem is ill-posed and on this account some regularization scheme has to be applied. We developed an inverse solver based on the Gauss-Newton-one-step method for differential imaging, and we implemented and tested four different regularization schemes: the first and second approaches employ a classical smoothness criterion using the unit matrix and a differential matrix of first order as the regularization matrix. The third method is based on variance uniformization, and the fourth method is based on the truncated singular value decomposition. Reconstructions were carried out with synthetic measurement data generated with a spherical perturbation at different locations within a conducting cylinder. Data were generated on a different mesh and 1% random noise was added. The model contained 16 excitation coils and 32 receiver coils which could be combined pairwise to give 16 planar gradiometers. With 32 receiver coils all regularization methods yield fairly good 3D-images of the modelled changes of the conductivity distribution, and prove the feasibility of difference imaging with MIT. The reconstructed perturbations appear at the right location, and their size is in the expected range. With 16 planar gradiometers an additional spurious feature appears mirrored with respect to the median plane with negative sign. This demonstrates that a symmetrical arrangement with one ring of planar gradiometers cannot distinguish between a positive conductivity change at the true location and a negative conductivity change at the mirrored location.

Algorithms↗
PubMed2006

Disentangling information flow in the Ras-cAMP signaling network.

The perturbation of signal-transduction molecules elicits genomic-expression effects that are typically neither restricted to a small set of genes nor uniform. Instead there are broad, varied, and complex changes in expression across the genome. These observations suggest that signal transduction is not mediated by isolated pathways of information flow to distinct groups of genes in the genome. Rather, multiple entangled paths of information flow influence overlapping sets of genes. Using the Ras-cAMP pathway in Saccharomyces cerevisiae as a model system, we perturbed key pathway elements and collected genomic-expression data. Singular value decomposition was applied to separate the genome-wide transcriptional response into weighted expression components exhibited by overlapping groups of genes. Molecular interaction data were integrated to connect gene groups to perturbed signaling elements. The resulting series of linked subnetworks maps multiple putative pathways of information flow through a dense signaling network, and provides a set of testable hypotheses for complex gene-expression effects across the genome.

Cyclic AMP↗
PubMed2005

Prenatal restraint stress and long-term affective consequences.

Chronic or repeated stress during critical periods of human fetal brain development has been associated with various learning, behavioral and/or mood disorders in later life. In this investigation, pregnant Fischer 344 rats was individually restrained three times a day for 45 min during the last week of gestation in transparent plastic cylinders while at the same time being exposed to bright light. Control pregnant females were left undisturbed in their home cages. Anxiety and depressive-like behavior was measured in the offspring at an age of 6 months using the open field test, the home cage emergence test and the forced swim test. Prenatally stressed rats spent more time in the corners and less time along the walls of an open field, while no difference in total distance moved was observed. In addition, prenatally stressed rats took more time to leave their home cage in the home cage emergence test. On the other hand, no differences in immobility were observed in the forced swim test. Moreover, prenatally stressed rats showed lower stress-induced plasma corticosterone levels compared with control rats. Prenatal stress (PS) had no effect on the number of 5-bromo-2-deoxyuridine-positive cells - used as a measure for cell proliferation - in the dentate gyrus of these rats. These data further support the idea that PS may perturb normal anxiety-related development. However, the present data also suggest that an adaptive or protective effect of PS should not be ignored. Genetic factors are likely to play a role in this respect.

Adaptation, Physiological↗
PubMed2003

Study of the competitive isotherm model and the mass transfer kinetics for a BET binary system.

The competitive adsorption behavior of the binary mixture of phenetole (ethoxy-benzene) and propyl benzoate in a reversed-phase system was investigated. The adsorption equilibrium data of the single-component systems were acquired by frontal analysis. The same data for binary mixtures were acquired by the perturbation method. For both compounds, the single-component isotherm data fit best to the multilayer BET model. The experimental overloaded band profiles are in excellent agreement with the profiles calculated with either the general rate model or the modified transport-dispersive models. The competitive adsorption data were modeled using the ideal adsorbed solution (IAS) theory. The numerical values of the coefficients were derived by fitting the retention times of the perturbation pulses to those calculated using the IAS theory compiled with the coherence conditions. Finally, the elution profiles of binary mixtures were recorded. They compared very well with those calculated. As a characteristic feature of this case, an unusual retainment effect of the chromatographic band of the more retained component by the less retained one was observed. The combination of the General Rate Model and the adsorption isotherm model allowed an accurate prediction of the band profiles.

Adsorption↗
PubMed1995

The influence of target perturbation on manual aiming asymmetries in right-handers.

Ten right-handed subjects performed 100 target-aiming movements with each hand. These movements were directed toward a small target on the midline. On 60% of the trials, the target remained stationary. On other randomly placed trials, the target "jumped" to a location 3 cm to the right (20%) or left (20%) of its original position when the cursor had travelled 6.5 cm. Although no hand differences were evident in the control condition, the right hand acquired the new target location more quickly than the left hand when the target was perturbed in either direction. Kinematic data revealed that this advantage was not due to initiating an adjustment to the initial movement more rapidly, but rather less time decelerating the corrective movement. Movement adjustments on perturbed trials were implemented more rapidly in left space than right space independent of the hand doing the aiming. These asymmetries may reflect the differential role of the two cerebral hemispheres in the control of goal-directed movements.

Female↗
PubMed2005

Characterization of female breast lesions from multi-wavelength time-resolved optical mammography.

Characterization of both malignant and benign lesions in the female breast is presented as the result of a clinical study that involved more than 190 subjects in the framework of the OPTIMAMM European project. All the subjects underwent optical mammography, by means of a multi-wavelength time-resolved mammograph, in the range 637-985 nm. Optical images were processed by applying a perturbation model, relying on a nonlinear approximation of time-resolved transmittance curves in the presence of an inclusion, with the aim of estimating the major tissue constituents (i.e. oxy- and deoxy-haemoglobin, lipid and water) and structural parameters (linked to dimension and density of the scatterer centres) for both the lesion area and the surrounding tissue. The critical factors for the application of the perturbation model on in vivo data are also discussed. Forty-six malignant and 68 benign lesions were analysed. A subset of 32 cancers, 40 cysts and 14 fibroadenomas were found reliable for the perturbation analysis. For cancers, we show a higher blood content with respect to the surrounding tissue, while cysts are characterized by a lower concentration of scattering centres with respect to the surrounding tissue. For fibroadenomas, the low number of cases does not allow any definite conclusions.

Breast Cyst↗
PubMed2006

Inference of gene regulatory networks and compound mode of action from time course gene expression profiles.

MOTIVATION: Time series expression experiments are an increasingly popular method for studying a wide range of biological systems. Here we developed an algorithm that can infer the local network of gene-gene interactions surrounding a gene of interest. This is achieved by a perturbation of the gene of interest and subsequently measuring the gene expression profiles at multiple time points. We applied this algorithm to computer simulated data and to experimental data on a nine gene network in Escherichia coli. RESULTS: In this paper we show that it is possible to recover the gene regulatory network from a time series data of gene expression following a perturbation to the cell. We show this both on simulated data and on a nine gene subnetwork part of the DNA-damage response pathway (SOS pathway) in the bacteria E. coli. CONTACT: dibernardo@tigem.it SUPLEMENTARY INFORMATION: Supplementary data are available at http://dibernado.tigem.it

Algorithms↗
PubMed1991

Positional uncertainty in long-term memory.

Long-term memory for position was examined from the perspective of an immediate memory framework, the perturbation model of Estes (Estes, 1972; Lee & Estes, 1977, 1981). First, a simple version of the perturbation model is shown to provide a reasonable fit of previously reported long-term data (Nairne, 1990b), even though the perturbation idea was developed to explain the phenomena of immediate retention. Second, new results are reported that extend the application to multiple dimensions. Long-term memories for list and within-list position appear to mimic the classic patterns of immediate retention, in that both show bow-shaped serial position curves and error generalization gradients that are roughly symmetrical around the true serial position.

Adult↗
PubMed2004

Perturbation theory for dark solitons: inverse scattering transform approach and radiative effects.

A perturbation theory for dark solitons of the nonlinear Schrödinger equation is developed. The theory is based on the inverse scattering transform method. Equations describing dynamics discrete (solitonic) and continuous (radiative) scattering data in the presence of perturbations are derived for N-soliton case. Adiabatic equations for soliton parameters and the perturbation-induced radiative field are obtained. The problem of the absence of a threshold for the creation of dark solitons under the action of a perturbation is discussed. A temporal one-soliton pulse with random initial perturbation and a spatial soliton with linear gain and two-photon absorption are considered as examples of application of the developed theory.

Journal Article↗
PubMed1982

A critique of the practice of plotting data obtained in vivo on an 'hours after treatment' format.

The erroneous and misleading conclusions that result when data obtained in vivo are plotted on an 'hours after treatment' format when compared to a 'time of day' or 'time of circadian period' format are illustrated. The mitotic index of the corneal epithelium and the amount of DNA synthetic activity in the tip of the tongue are the examples used. Animals were kept on a light-dark cycle with light from 06.00 to 18.00 CST. Treatment with an intraperitoneal injection of 0.2 ml of saline at 05.00 compared to treatment with saline at 17.00 resulted in no effect on either variable when the data were plotted on a 'time of day' format, i.e., the data from the group which received saline at 05.00 and the group which received saline at 17.00 were very reproducible. However, when the same data were plotted on the 'hours after treatment' format, the data were 180 degrees out of phase with each other. This resulted in many statistically significant differences between the two groups. These differences are artifactual when compared to the 'no effect' or 'no perturbation' situation seen when the data are plotted on the 'time of day' format.

Animals↗