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Biomedical subjects

H H Nguyen

Publications and source records attributed to H H Nguyen.

15 recordsLinked to original sources

Spectroscopic characterization of heme A reconstituted myoglobin.

The focus of this study was to examine the functional role of the unusual peripheral substitution of heme A. The effects of heme A stereochemistry on the reconstitution of the porphyrin have been examined in the heme A-apo-myoglobin complex using optical absorption and resonance Raman and electron paramagnetic resonance spectroscopies. The addition of one equivalent of heme A to apo-Mb produces a complex which displays spectroscopic signals consistent with a distribution of high- and low-spin heme chromophores. These results indicate that the incorporation of heme A into apo-Mb significantly perturbs the protein refolding.

Animals

Contemporary evaluation of unilateral vocal cord paralysis.

Unilateral vocal cord paralysis is a common finding in the practice of otolaryngology. Multiple etiologies have been described and have not changed appreciably in the last century. We attempted to characterize the contemporary evaluation of unilateral vocal cord paralysis, with consideration given to cost-effectiveness. Thirty-one board-certified otolaryngologists were interviewed to determine their typical evaluation protocol. The average cost of an evaluation totaled $1706.18, with a range of $112.56 to $3439.52. Otolaryngologists with more years of experience tended to pursue briefer and less expensive evaluations. The charts of 187 patients with a diagnosis of vocal paralysis from 1983 to 1991 were reviewed, of which 113 were evaluable. Eighty-four of these 113 (74%) were unilateral. In 48 of 84 cases (57%), the cause was apparent at the time of diagnosis. In 36 of 84 cases (43%), an evaluation was necessary. A diagnosis was achieved in 27 of these 36 instances (75%), with the most useful test being a chest roentgenogram (n = 13, 48%). The most common cause of unilateral vocal cord paralysis in our series was neoplasm (n = 34, 40%), followed by surgical trauma (n = 29, 35%). In no instance was a malignancy discovered subsequent to the initial evaluation. The most cost-efficient, inclusive diagnostic evaluation of unilateral vocal cord paralysis involves a stepwise progression through the tests that are most likely to yield a diagnosis, with endoscopy reserved for those cases in which simpler, less invasive tests have not indicated a cause.

Adult

Degradation of glomerular basement membrane by purified mammalian metalloproteinases.

Neutral metalloproteinases degrade components of the extracellular matrix, including collagen types I-V, fibronectin, laminin and proteoglycan. However, their ability to degrade intact glomerular basement membrane (GBM) has not previously been investigated. Incubation of [3H]GBM (50,000 c.p.m.; pH 7.5; 24 h at 37 degrees C) with purified gelatinase or stromelysin (2 units) resulted in significant GBM degradation: gelatinase, 46 +/- 2.2; stromelysin, 59 +/- 5.8 (means +/- S.E.M.; percentage release of non-sedimentable radioactivity; n = 4). In contrast, 2 units of collagenase released only 5.6 +/- 0.52% (n = 3) of the [3H]GBM radioactivity compared with 2.0 +/- 0.15% (n = 7) released from [3H]GBM incubated alone. Sephadex G-200 gel chromatography of supernatants obtained from incubations of [3H]GBM with either gelatinase or stromelysin confirmed the ability of these enzymes to degrade GBM and revealed both high-(800,000) and relatively low-(less than 20,000) Mr degradation products for both enzymes. GBM degradation by gelatinase and stromelysin was dose-dependent (range 0.02-2.0 units), near maximal between pH 6.0 and 8.6, and was completely inhibited (greater than 95%) by 2 mM-o-phenanthroline. Collagenase (2 units) did not enhance the degradation of GBM by either gelatinase (0.02 or 0.2 unit) or stromelysin (0.02 or 0.2 unit). Our results indicate that metalloproteinase-mediated GBM degradation by neutrophils and glomeruli may be attributable to gelatinase (neutrophils) and/or stromelysin (glomeruli) and suggest an important role for these proteinases in glomerular pathophysiology.

Animals

Pathology of the cardiac conduction system in myotonic dystrophy: a study of 12 cases.

In 12 autopsy cases of myotonic dystrophy, the most frequently observed histopathologic lesions of the cardiac conduction system were fibrosis, fatty infiltration and atrophy. Fibrosis involved the sinus node in 6 cases, atrioventricular (AV) node in 7, AV bundle in 8, bundle branches in 10 and ventricular myocardium in 11. Fatty infiltration was observed in the sinus node in two cases, AV node in two, AV bundle in six, bundle branches in one and ventricular myocardium in nine. Atrophy was prominent in the AV bundle in five and bundle branches in eight. Lymphocytes infiltrated the conduction system in three cases and were associated with myotonic dystrophy in two and varicella myocarditis in one. Ventricular myocytes were hypertrophied in seven cases, vacuolated in three and exhibited disarray in two. The distribution and extent of conduction system lesions tended to correspond to antemortem electrocardiographic abnormalities, including prolonged PR interval in six cases, intraventricular conduction delay in six and bundle branch block in four. Cardiac involvement by myotonic dystrophy may have contributed to sudden death in four cases.

Adolescent

Aldosterone and dexamethasone binding in human arterial smooth muscle cells.

Regulation of blood pressure by direct action of corticosteroids on blood vessel walls was first hypothesized in 1952 [1]. The presence of receptors specific for these hormones within the peripheral vessel walls is a pre-requisite of this hypothesis. This report documents specific binding of both aldosterone (AL) and dexamethasone (DM) in cultured human arterial smooth muscle cells. After the arterial smooth muscle cultures were incubated with tritiated AL or DM at 37 degrees C for 30-60 min, the cells were sonicated and the protein bound steroid fraction isolated on a Sephadex G25 column. Using ion exchange chromatography of this fraction, each corticosteroid receptor complex displayed a distinct, reproducible elution pattern. Aldosterone showed a single peak at 0.096 +/- 0.005 mol/l sodium phosphate and DM had two peaks at 0.029 +/- 0.003 and 0.050 +/- 0.004 mol/l sodium phosphate. The Scatchard plots of specific binding from AL saturation curves are linear and revealed mean +/- s.d. steady state binding parameters of dissociation constant (Kd) = 0.35 +/- 0.15 nmol/l and maximum binding capacity (Bmax) = 98 +/- 53 X 10(-18) mol/micrograms DNA. Similarly, the mean +/- s.d. steady state binding parameters for DM are Kd = 4.4 +/- 2.0 nmol and Bmax = 3031 +/- 1385 X 10(-18) mol/micrograms DNA. Therefore, there are approximately 1150 AL binding sites and 30000 DM binding sites per cell. The Kd and Bmax values are similar to those previously described for corticoid receptors in rat aortic smooth muscle cells and are consistent with physiological steroid concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Degradation of glomerular basement membrane by a neutral metalloproteinase(s) present in glomeruli isolated from normal rat kidney.

Incubation of glomerular homogenates (200 micrograms protein) with glomerular basement membrane (GBM, 30-35 micrograms hydroxyproline) at pH 7.5 for 36 h at 37 degrees C resulted in significant GBM degradation as measured by hydroxyproline release (40 +/- 6%, n = 17). GBM degradation increased with increasing incubation time (12-48 h) and glomerular protein concentration (50-250 micrograms). GBM degradation was not significantly decreased by inhibitors of serine or cysteine proteinases or the inhibitor of bacterial metalloproteinases, phosphoramidon. In contrast GBM degradation by glomerular homogenates was markedly inhibited by the metal chelators 10mM EDTA (-95 +/- 3%, n = 7) and 2mM 1,10-phenanthroline (-96 +/- 2%, n = 4). Preincubation of glomerular homogenates with trypsin (followed by soya bean trypsin inhibitor) markedly stimulated GBM degradation (+103 +/- 20%, n = 11). These results document the presence of a GBM-degrading, neutral metalloproteinase(s) in glomeruli suggesting an important role for this enzyme in glomerular pathophysiology.

Animals

A physico-chemical comparison of aortic receptors in rat hypertension models.

Rat models of genetic hypertension include spontaneous hypertension and resistance or sensitivity to mineralocorticoid and salt induced hypertension. Previously, altered aldosterone binding to corticoid receptor I was found in aortic smooth muscle cells cultured from Fischer 344 rats which are extremely resistant to steroid and salt induced hypertension. The corticoid receptor I of Fisher 344 rats had a lower affinity than that of salt sensitive Wistar-Kyoto controls, as well as spontaneously hypertensive rats and Sprague-Dawley rats. In the present study, we have used DEAE-cellulose ion exchange chromatography to compare the structure (charge properties) and steroid specificity of vascular corticoid receptor I and II sites in these same rat hypertension models. No variations in ion exchange properties of type I and II receptors were found. Together with the lower aldosterone affinity of corticoid receptor I sites in Fischer 344 rats these data suggest an altered binding domain which is not seen as a difference in charge density of the receptor protein by ion exchange chromatography.

Aldosterone

Physical separation of aortic corticoid receptors with type I and type II specificities.

Previous gel filtration binding assay studies indicated that rat vascular smooth muscle cells contained corticoid receptor I and corticoid receptor II sites which could be distinguished on the basis of their relative affinities for aldosterone and dexamethasone. Ion-exchange chromatography experiments were designed to separate the two sites for further studies on their physical characteristics and role in vascular smooth muscle cell physiology. Cultured aortic cells were incubated with 5-10 nM 3H steroid alone or in the presence of 10-fold non-radioactive steroid competitor for 30 min at 37 degrees C. Following cell lysis, total cellular protein-bound steroid was isolated using Sephadex G-25 and applied to a DEAE-cellulose ion-exchange column. Three peaks of radioactivity were eluted using a 1-200 mM sodium phosphate gradient: peak I (30-38 mM), peak II (52-64 mM), and peak III (92-102 mM). Peaks I and II contained 60% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor II sites (dexamethasone greater than aldosterone). Peak III contained 40% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor I sites (aldosterone greater than dexamethasone). These studies support the binding assay data on steroid specificity and relative proportion of type I and II sites. They also document the existence of type I and II corticoid receptors with different physicochemical characteristics in rat aortic smooth muscle cells.

Aldosterone

Extra-high-frequency noise remotely masks and alters temporal integration at lower frequencies.

A quasi-free-field technique was used to assess the effects of extra-high-frequency-band masking on detection threshold of middle and high (1-7 kHz) frequencies. At an SPL of 60 dB, the 10- to 20-kHz masker produced a slight amount of masking. Increasing the masking level by another 5 dB produced a disproportionate increase of the detection thresholds. This increase was greatest in response to 4- and 5-kHz stimuli, and was detected with both constant and pulsed tones. Decreasing the bandwidth of the masker reduced the magnitude of the effect, but not the frequency-specific pattern of the remote masking. Extra-high-frequency masking also increased the amount of temporal integration at middle and high frequencies. The data are discussed in reference to peripheral and central neural changes associated with sensorineural hearing loss.

Adult

Extra-high-frequency auditory thresholds: fine structure, reliability, temporal integration and relation to ear canal resonance.

Three trained subjects were repeatedly tested for detection of auditory thresholds at 27 frequencies, from 2 to 18 kHz. A double-blind procedure was used with the method of adjustment, and the quasi-free-field delivery system of Osterhammel et al. [Scand. Audiol. 6:91-95, 1977] was used to monaurally stimulate the ear. A very reliable series of high- and low-threshold changes, termed the fine structure of the extra-high-frequency audiogram, was observed from 8 to 16 kHz. All 6 ears had a major low-threshold region at or near 13.5 kHz, and up to 3 other replicable sensitive frequencies along this one octave range. Thresholds were invariant over 3 conditions of stimulus presentation (constant tone; 500 ms duration, 25 ms rise and decay, 1 s interstimulus interval; and 100 ms duration, 1.5 ms rise and decay, 200 ms interstimulus interval). As the duration of the tone was decreased from 100 to 5 ms, temporal integration (an average of approximately 2.5 dB threshold increase as the duration was halved) was noted. Although temporal integration was slightly less at higher frequencies, the shape of the audiograms remained essentially unchanged as the stimulus duration was decreased to 5 ms. There was no consistent relationship between the maximally sensitive high frequencies and the amount of temporal integration. But there was a general association between the maximally sensitive high frequencies and the resonant frequency characteristics of that particular ear.

Adult

Rosette-forming cells in patients with treated leprosy.

White blood cell counts and the percentages and absolute numbers of lymphocytes in the peripheral blood of active lepromatous (BT, BB, BL) leprosy patients, patients with borderline leprosy, and normal controls were determined. Lepromatous patients showed decreased leukocyte counts and elevated percentages of lymphocytes, resulting in normal absolute lymphocyte counts. The proportion of peripheral blood mononuclear cells forming "active" rosettes, standard (4 degrees C overnight) rosettes, and "high affinity" (29 degrees C for 1 hr) rosettes with sheep erythrocytes, and rosettes with EAC were determined. Lepromatous patients, compared with normal controls, had decreased "active" rosettes, standard rosettes, and "high affinity" rosettes with sheep erythrocytes with an increase in the nonrosetting proportion. Both lepromatous and borderline leprosy patients showed increased percentages of EAC rosettes compared with normal controls.

Humans