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PubMed2006

Glycosylation of recombinant proforms of major house dust mite allergens Der p 1 and Der f 1 decelerates the speed of maturation.

BACKGROUND: The efficient manufacture of recombinant Der p 1 and Der f 1 has been an important bottleneck in the study of house dust mite allergies and the development of applications for allergen engineering. While Der f 1 has only one N-glycosylation motif in the mature sequence, Der p 1 has two motifs, one in the prosequence and the other in the mature sequence. To test whether inefficient maturation of a recombinant Pro-Der p 1 versus Pro-Der f 1 is due to N-glycosylation, the maturation speed of N-glycosylation motif mutants was compared. METHODS: Expression vectors for the mutants, in which the motif in the Der p 1 prodomain was disrupted or a motif was created within the Der f 1 prodomain, were constructed by site-directed mutagenesis of preproforms with or without the motif within the mature portion. Culture supernatants of yeast Pichia pastoris transfectant cells containing proforms were buffer exchanged by gel filtration and incubated for maturation. Samples from the reactions were collected every 20 min and subjected to electrophoresis. The maturation speed was compared based on the band densities of the pro- and mature forms. RESULTS: Disruption of the motif in the mature portion decreased the productivity and accelerated the maturation. Maturation was also accelerated by disruption of the other motif in the Der p 1 prodomain and slowed down by introduction of the motif into the Der f 1 prodomain. CONCLUSIONS: Maturation systems using Pro-Der p 1 without the prodomain glycosylation are useful for the efficient preparation of a recombinant mature allergen. In addition, these results demonstrated that the maturation of cysteine protease could be controlled through glycosylation of the prodomain.

Amino Acid Sequence↗
PubMed1990

Growth response to growth-hormone administration during the decelerating phase of the pubertal growth spurt in short normal children.

In order to investigate the value of growth hormone (GH) treatment during late puberty, we studied the effect of human GH (hGH) administration (0.85 +/- 0.30 IU/kg/week; range: 0.44-1.28) on height velocity (HV) after the peak of the pubertal growth spurt in a group of 10 (4 girls and 6 boys) short normal children (GH peak after pharmacological stimulation: 15.5 +/- 2.3 ng/ml) with growth retardation (height: 2.6 +/- 0.3 SD) and puberty Tanner stage 4. A group of 10 untreated children, observed prior to the study, served as controls. The children were regularly measured during their pubertal growth spurt, and HV (cm/year) was calculated every 6 months. The pretreatment evaluation consisted of 2 consecutive 6-month periods characterized by a decrease in HV of at least 25%. In the group of selected children, hGH administration was then initiated and growth variables were evaluated after 6 and 12 months of therapy. Skeletal maturation was evaluated at the beginning as well as after 6 months and 12 months of hGH therapy. In the controls, HV (mean +/- SD) had decreased from 8.8 +/- 1.8 to 4.9 +/- 1.4 cm/year during the pretreatment period (in girls from 7.9 +/- 1.4 to 4.1 +/- 0.6 cm/year and in boys from 9.6 +/- 1.6 to 5.8 +/- 1.2 cm/year). During the following semester, HV was 3.3 +/- 0.8 cm/year (girls: 3.4 +/- 1.0 and boys: 3.2 +/- 0.2 cm/year).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed2006

State psychiatric hospital census after the 1999 Olmstead Decision: evidence of decelerating deinstitutionalization.

OBJECTIVE: The Supreme Court ruled in the 1999 Olmstead decision that "unjustified isolation" of individuals with disabilities in institutions is a violation of the Americans With Disabilities Act. This study examined the extent to which state psychiatric hospital census across the United States has changed significantly post-Olmstead. METHODS: Twenty years of national state hospital census data (1984-2003) were used to assess trends in the rate of declines from pre- to post-Olmstead periods. Data were organized into five four-year periods. RESULTS: Steady declines in the hospital census nationally were found over all periods, with especially large decreases in the 1990s. However, when the percent change in hospital census in the two periods immediately before the Olmstead decision (between 1992-1995 and 1996-1999) were compared with the percent change in the periods immediately before and immediately after the Olmstead decision (between 1996-1999 and 2000-2003), an 8 percent decrease in the magnitude of decline was seen. CONCLUSIONS: State hospital census continues to decline but has slowed significantly during the post-Olmstead period. More study of the factors associated with this decline is needed.

Censuses↗
PubMed1984

Evidence from the rhesus monkey (Macaca mulatta) for the view that negative feedback control of luteinizing hormone secretion by the testis is mediated by a deceleration of hypothalamic gonadotropin-releasing hormone pulse frequency.

The site and mode of the feedback actions of testicular hormones on gonadotropin secretion in the adult rhesus monkey were investigated using the arcuate-lesioned preparation previously employed by others to study cognate problems in the female. The negative feedback loop that governs LH and FSH release in the male monkey was opened without changing either the frequency or amplitude of intermittent GnRH stimulation of the pituitary gonadotrophs, which was clamped by exogenous GnRH replacement at a level that approximated the intact or closed loop hypophysiotropic signal. In this manner, the relative importance of adenohypophysial vs. hypothalamic sites of feedback action of testicular hormones on LH and FSH secretion was assessed. To accomplish the foregoing, radiofrequency lesions were placed in the region of the arcuate nucleus to abolish endogenous hypothalamic GnRH secretion. Patterns of temporally coupled episodes of pituitary LH and testicular testosterone discharge that in nonlesioned animals characteristically occur, on the average, once every 3 h throughout the 24-h light-dark cycle were restored in lesioned animals by an intermittent iv infusion of GnRH (0.1 micrograms/min for 3 min every 3 h). Bilateral orchidectomy in this experimental paradigm elicited only small increments in LH pulse amplitude and mean plasma LH concentration, a response in striking contrast to the dramatic postcastration LH hypersecretion observed in animals with intact hypothalami that respond to the opening of the negative feedback loop with an apparent acceleration in the endogenous frequency of intermittent GnRH secretion. A marked rise in mean plasma LH concentration in arcuate-lesioned males, however, was forth-coming when the frequency of intermittent exogenous GnRH stimulation was increased 2-3 weeks after castration from one pulse every 3 h (intact frequency) to one pulse per h (castrate frequency). These findings fail to provide evidence for a major inhibitory feedback action of the testes on LH secretion at the level of the adenohypophysis. They are entirely consistent, however, with the hypothesis that the negative feedback control of LH release by the male gonad is mediated, principally, via the central nervous system by an action of testicular hormone, most probably testosterone, to retard the frequency of the neural timing mechanism that governs the intermittent pattern of GnRH release by the hypothalamus.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2000

Exendin-4 decelerates food intake, weight gain, and fat deposition in Zucker rats.

Exendin-4 is a 39 amino acid peptide produced in the salivary gland of the Gila monster lizard. It has a 53% amino acid homology to the incretin hormone glucagon-like peptide-1 (GLP-1). Exendin-4 induces insulin release through activation of the GLP- 1 receptor but is a much more potent insulinotropic agent than GLP-1. Of critical importance for its potential use as a treatment for diabetes is its much longer biological effect in vivo. Previous studies involving once daily administration of exendin-4 over 13 weeks to db/db mice demonstrated that it lowers hemoglobin A1c (HbA1c), a marker of mean blood glucose levels. Food consumption in the treated animals dropped over the first 4 days and then increased to a level comparable with that of the untreated animals. In this study, we initially examined the effect of once daily injections (over 14 days) on the food consumption of Zucker fatty rats. We observed an immediate reduction in food intake which then leveled off(after 5 days) to match that of the untreated animals. Subsequently we injected the same animals twice daily (treatment period of 56 days in total) and observed a sustained reduction in food intake and weight-gain. This was matched by a reduction in the critical parameters of HbA1c, fasting blood glucose and plasma insulin. MRI imaging of the abdominal regions of the animals showed that initially only the amount of fat deposited in the sc region was reduced after 4 weeks exendin-4 treatment. At the 8-week time point there was a corresponding decrease in the amount of visceral fat deposition. The combination of appetite reduction, decreased fat deposition and an improvement in the parameters associated with glucose intolerance makes a case for the use of exendin-4 as a treatment for diabetes.

Adipose Tissue↗
PubMed1998

Deceleration by angiotensin II of the differentiation and bone formation of rat calvarial osteoblastic cells.

We examined the effects of angiotensin II (Ang II) on the differentiation of rat calvarial osteoblastic cells and on the formation of bone by these cells. Northern blotting analysis revealed that Ang II inhibited the expression of mRNA for osteocalcin, which is a protein that is specifically expressed during maturation of osteoblastic cells. Ang II decreased the activity of alkaline phosphatase, a marker of osteoblastic differentiation, in the cells, acting via the type 1 (AT1) receptor. We used von Kossa staining to examine the formation of mineralized nodules by osteoblastic cells. Both the number and the total area of mineralized nodules were quantified and shown to be decreased by 10(-7) M Ang II. The accumulation of calcium in cells and the matrix layer was also decreased by Ang II. Binding analysis with subtype-specific antagonists revealed the presence of AT1 receptors for Ang II in this culture system. Ang II caused a marked increase in the rate of production of intracellular cAMP in this system. Our data suggest that Ang II might be intimately involved in osteoblastic metabolism through its interaction with the AT1 receptor.

Alkaline Phosphatase↗
PubMed1979

Cardiac deceleration and accuracy of performance: recalculation of standard score transform.

In two previous studies the standard score transform used was based on a standard deviation derived from the mean standard deviations of the raw data in the six classifications employed in the studies rather than on one based on the entire set of raw scores as is the more common practice. The data were reanalyzed using the latter basis and major findings were confirmed. Some of the minor findings, however, were changed. These changes are pointed out and discussed where they occurred.

Achievement↗
PubMed1982

Decelerated synthesized speech as a means of shaping speed of auditory processing of children with delayed language.

This study investigated whether the rate at which children with delayed language process auditory stimuli can be increased through a process of shaping with synthesized speech stimuli. Results indicate that stimuli with slowed down critical formant cues were easier for these 24 children with delayed language to discriminate and that training with stimuli of extended duration did generalize to stimuli of normal durations. Implications of these findings and usefulness of this methodology are discussed.

Child↗