Longitudinal exposure to antiseizure medications shape gut-derived microbiome, resistome, and metabolome landscape.

The influence of chronically administered host-targeted drugs on the gut microbiome remains less understood compared to antibiotics. We investigated repetitive exposure effects of three common antiseizure medications [carbamazepine (CBZ), valproic acid, and levetiracetam] on the gut microbial composition, resistome, and metabolome using microcosms constructed from feces of young children. Microcosms were established by cultivating feces for 24 h (C0).

Syndrome of coronal craniosynostosis with brachydactyly and carpal/tarsal coalition due to Pro250Arg mutation in FGFR3 gene.

Recently, a unique Pro250Arg point mutation in fibroblast growth factor receptor 3 (FGFR3) was reported in 61 individuals with coronal craniosynostosis from 20 unrelated families [Muenke et al. (1997): Am J Hum Genet 60:555-564]. The discovery of this apparently common mutation has resulted in the definition of a recognizable syndrome, through analysis of subtle clinical findings in families who were previously thought to have a variety of other craniosynostosis syndromes.

Pseudohypoparathyroidism type Ia from maternal but not paternal transmission of a Gsalpha gene mutation.

While loss-of-function mutations in Gsalpha are invariably associated with the short stature and brachydactyly of Albright hereditary osteodystrophy (AHO), the association with hormone resistance (to parathyroid hormone and thyrotropin) typical of pseudohypoparathyroidism type Ia (PHP-Ia) is much more variable. Observational studies and DNA polymorphism analysis suggest that maternal transmission of the Gsalpha mutation may be required for full expression of clinical hormone resistance. To test this hypothesis, we studied transmission of a frameshift mutation in Gsalpha through three generations of a pedigree affected by AHO and PHP-Ia.

Real-time monitoring of reduced beta-adrenergic response in fibroblasts from patients with pseudohypoparathyroidism.

The activation of cell-surface receptors usually produces a transient increase of the extracellular acidification rate in culture cells that can be detected with a biosensor-based instrument. We describe here the application of this method in monitoring hormonal response of Galphas-deficient fibroblasts from patients with pseudohypoparathyroidism type Ia (PHP-Ia). We found that following exposure to isoproterenol, the mean acidification response of fibroblasts from four PHP-Ia patients was only 18% of the response in normal cells.

Concurrent hormone resistance (pseudohypoparathyroidism type Ia) and hormone independence (testotoxicosis) caused by a unique mutation in the G alpha s gene.

Defects in the G (guanine nucleotide-binding)-protein subunit (G alpha s) which stimulates adenylyl cyclase may result in either loss or gain of endocrine function. Reduced G alpha s activity is found in the hormone resistance syndrome, pseudohypoparathyroidism type Ia (PHP-Ia), while constitutive activation of G alpha s is associated with endocrine organ overactivity, including the gonadotropin-independent sexual precocity seen in patients with McCune-Albright syndrome. We identified two unrelated boys presenting with concurrent PHP-Ia and gonadotropin-independent sexual precocity (testotoxicosis).

IGF-I resistance in virus-transformed B-lymphocytes from African Efe Pygmies.

To investigate IGF-I resistance in African Efe Pygmies, we examined clonal responsiveness to IGF-I in Epstein-Barr virus-transformed B-lymphocytes from three Efe Pygmies and three American control subjects. The Efe B-lymphoblasts did not increase clonal responsiveness when incubated with IGF-I (as high as 250 micrograms/liter) in contrast to the control B-lymphoblasts which showed a bimodal dose-response with a maximal stimulation of 50% above baseline. The proliferative response of Efe B-lymphoblasts was similar to that of control B-lymphoblasts when incubated with another growth factor, phorbol 12-myristate 13-acetate, which does not activate the IGF-I receptor.

Decreased cortical and increased cancellous bone in two children with primary hyperparathyroidism.

The basis for this study is two children with primary hyperparathyroidism (PHPT) who radiographically manifested both marked subperiosteal resorption and prominent osteosclerosis. We hypothesize that the parathyroid hormone (PTH) elevation not only increased osteoclastic resorption of cortical bone but also simultaneously enhanced cancellous bone formation, giving rise to osteosclerosis. In this report, we describe the changes in trabecular and cortical bone density, as measured by quantitative computed tomography (QCT), in these two young patients with severe PHPT, before and after removal of a parathyroid adenoma.

Steady-state mRNA levels of G protein subunits in developing rabbit myocardium.

Cardiac responsiveness to beta-adrenergic stimulation changes with age. Developmental changes in expression of guanine nucleotide-binding coupling protein (G protein) subunits may account for these physiologic changes. We measured steady-state levels of mRNA encoding the alpha-subunit of the specific G protein that stimulates adenylyl cyclase (Gs alpha) and three isoforms of beta-subunit of G proteins (G beta) in developing myocardium.

Adenylyl cyclase integrates multiple G protein signals to modulate calcium currents in neonatal rabbit heart.

We investigated the effects of added beta gamma subunits of G proteins (G beta gamma) on beta-adrenergic responsiveness of transmembrane Ca2+ currents (ICa) in ventricular myocytes from neonatal rabbits. G beta 1 gamma 1 purified from retinal rods was dialyzed into cells via the voltage clamp micro-electrode. Stimulation of ICa by isoproterenol was not affected by added intracellular G beta 1 gamma 1 or by carbachol alone but was completely blocked by combined G beta 1 gamma 1 and carbachol.

The HLA-A3, Cw6,B47,DR7 extended haplotypes in salt losing 21-hydroxylase deficiency and in the Old Order Amish: identical class I antigens and class II alleles with at least two crossover sites in the class III region.

The HLA-B47,DR7 haplotype in congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency contains a deletion of most of the active CYP21 gene and the entire adjacent C4B gene. The C4A gene produces a protein which is electrophoretically C4A but antigenically C4B. In the Old Order Amish, the HLA-B47,DR7 haplotype contains no deletion, but is immunologically identical to the CAH haplotype in both areas flanking the crossover region.

Developmental changes in mRNA encoding cardiac Na+/H+ exchanger (NHE-1) in rabbit.

The faster recovery of cardiac contractility in newborn rabbit hearts during acute acidosis compared to adult hearts correlates with greater cellular activity of the Na+/H+ exchanger. We quantified mRNA encoding Na+/H+ exchanger-1 (NHE-1) in rabbit ventricles of fetal (27 days gestation), newborn (2-5 days), and adult (> 6 months) New Zealand white rabbits using reverse transcription-polymerase chain reaction (RT-PCR) and RNase protection assay, with GAPD mRNA as standard. Both RT-PCR and RNase protection assay revealed similar (p > 0.

Effects of triiodothyronine on retention of beta-adrenergic responsiveness of voltage-gated transmembrane calcium current during culture of ventricular myocytes from neonatal rabbits.

To study the effects of triiodothyronine (T3) on responsiveness of L-type calcium currents to beta-adrenergic stimulation in neonatal hearts, ventricular myocytes were isolated from neonatal rabbits and cultured in medium containing 10% fetal bovine serum to which T3 had been added to achieve either hypothyroid, euthyroid, or hyperthyroid conditions, as assessed by measurement of free T3 concentrations. During a 24-h culture period, the striated rod-shaped myocardial cells progressively assumed a stellate shape with reduced surface area; however, the rate constants for diffusion of Na+ from a microelectrode pipette into the cells remained unchanged. Voltage-dependent characteristics of L-type calcium currents as assessed by whole-cell voltage clamp studies were also unchanged after culture with various concentrations of free T3.

Characterization of heterogeneous mutations causing constitutive activation of the luteinizing hormone receptor in familial male precocious puberty.

Familial male precocious puberty (FMPP) is a gonadotropin-independent disorder that is inherited in an autosomal dominant, male-limited pattern. A heterozygous mutation encoding substitution of Asp578 with Gly in transmembrane helix 6 of the G protein-coupled receptor for luteinizing hormone (LHR) has been found in affected males from nine American FMPP families. Cells expressing the mutant LHR exhibit markedly increased cyclic adenosine monophosphate (cAMP) production in the absence of agonist, suggesting that autonomous Leydig cell activity in FMPP is caused by a constitutively activated LHR.

Rapid GDP release from Gs alpha in patients with gain and loss of endocrine function.

Luteinizing hormone stimulates testicular Leydig cells to produce testosterone by binding to a receptor that activates the G protein Gs and adenylyl cyclase. Testotoxicosis is a form of precocious puberty in which the Leydig cells secrete testosterone in the absence of luteinizing hormone, often due to constitutive activation of the luteinizing hormone receptor and (indirectly) Gs (refs 1-4). Here we study two unrelated boys suffering from a paradoxical combination of testotoxicosis and pseudohypoparathyroidism type Ia (PHP-Ia), a condition marked by resistance to hormones acting through cyclic AMP (parathyroid hormone and thyroid-stimulating hormone) as well as a 50% decrease in erythrocyte Gs activity (the remaining 50% is due to the normal Gs allele).

A method for isolation of rat renal microvessels and mRNA localization.

The objective of this study was to develop a technique to identify and dissect segments of the rat renal microcirculation and to apply reverse transcription (RT) to specific mRNAs with subsequent amplification of the cDNA by polymerase chain reaction (PCR) to evaluate gene expression. To circumvent the difficulty associated with visualizing specific microvessels, we intrarenally infused blue latex microparticles, 1-5 microns in diameter, with subsequent identification and microdissection of specific segments of the renal microvasculature under stereomicroscopy. To demonstrate its utility, we assessed expression of mRNAs encoding fibronectin and renin.

Growth hormone treatment of growth failure among children with renal transplants.

Eight children with growth failure following renal transplant have been selected for recombinant human growth hormone (rhGH) treatment at Children's Hospital using the following criteria: (1) a functioning allograft for at least one year; (2) height < third percentile; (3) growth velocity < 4 cm/year; (4) growth potential; and (5) low-dose alternate-day glucocorticoid dosing. The children were 7.4 to 17.

Congenital heart disease associated with sporadic Kallmann syndrome.

A 17-year-old boy with Kallmann syndrome had complex congenital heart disease that included double-outlet right ventricle, d-mal-position of the great arteries, right aortic arch, and hypoplastic main pulmonary artery. He had neurosensory hearing loss and mental retardation. The 7 previously reported patients with Kallmann syndrome and cardiac abnormalities were short with height > or = 2 standard deviations below the mean for age (5/7), lacked a family history of Kallmann syndrome (6/6), and were mentally retarded (4/4).

An arginine to histidine mutation in codon 311 of the C-erbA beta gene results in a mutant thyroid hormone receptor that does not mediate a dominant negative phenotype.

We have examined the c-erbA beta thyroid hormone receptor gene in a kindred, G.H., with a member, patient G.

Prevalence of three mutations in the Gs alpha gene among 24 families with pseudohypoparathyroidism type Ia.

Pseudohypoparathyroidism type Ia (PHP-Ia), an inherited multi-hormone resistance syndrome, is associated with deficient cellular activity of the alpha-subunit of the guanine nucleotide-binding protein (Gs alpha) that stimulates adenylyl cyclase. We determined prevalence of three recently described mutations in exons 1 and 10 of the Gs alpha gene among 24 unrelated patients with PHP-Ia. Restriction analysis was used to detect two mutations that produce unique RFLPs, and allele-specific oligonucleotide hybridization was used to detect the other mutation.

Accelerated growth rates in children treated with growth hormone after renal transplantation.

To determine the usefulness of growth hormone treatment among children with renal allografts, we treated nine children with functioning renal transplants who were less than 16 years of age and had poor growth. The nine children, who were aged 12.6 +/- 4.

Glucose tolerance in children with renal allografts and effect of growth hormone treatment.

We performed oral glucose tolerance tests (oGTTs) on 15 children who had functioning renal allografts received greater than or equal to 18 months previously, had adequate renal function, and had heights greater than 2.5 SD below the mean height for age. Three of the children had impaired glucose tolerance; their mean glucose levels during the last 2 hours of the oGTT were higher (p less than 0.

Immunodetectable levels of the inhibitory guanine nucleotide-binding regulatory proteins in failing human heart: discordance with measurements of adenylate cyclase activity and levels of pertussis toxin substrate.

Human hearts with idiopathic dilated cardiomyopathy have diminished adenylate cyclase activity and increased amounts of the alpha-subunit of the inhibitory guanine nucleotide-binding regulatory protein (alpha Gi) as measured by pertussis toxin catalyzed ADP-ribosylation. We utilized specific antisera against synthetic peptides corresponding to amino sequences deduced from cDNA's encoding the three alpha Gi subspecies to compare the immunologic and bioactivity levels of Gi in failing and non-failing human hearts. The various antisera detected three peptides with Mr 42,000, 38,000, and 37,000.

Diminished beta-adrenergic receptor responsiveness and cardiac dilation in hearts of myopathic Syrian hamsters (BIO 53.58) are associated with a functional abnormality of the G stimulatory protein.

Previous studies have demonstrated a diminution in the bioactivity of the guanine nucleotide-binding regulatory protein that stimulates adenylyl cyclase (Gs) in hearts of the hypertrophic BIO 14.6 Syrian hamster. In this study, we measured functional activity and immunodetectable levels of Gs in a mutant strain of hamsters (BIO 53.