Biomedical subjects

D L Miller

Publications and source records attributed to D L Miller.

At least 253 records · Page 14Linked to original sources

PubMed1991

Petrosal sinus sampling with and without corticotropin-releasing hormone for the differential diagnosis of Cushing's syndrome.

BACKGROUND: Measurement of adrenocorticotropin levels in plasma from the inferior petrosal sinuses of patients with Cushing's syndrome can distinguish adrenocorticotropin-secreting pituitary tumors (Cushing's disease) from other causes of the syndrome, principally ectopic adrenocorticotropin secretion from an occult tumor. However, it is unknown whether such measurement consistently identifies patients with Cushing's disease and whether testing with corticotropin-releasing hormone (CRH) enhances the value of the procedure. METHODS: We prospectively studied 281 patients with Cushing's syndrome to evaluate the diagnostic efficacy of the procedure. Bilateral sampling was successfully accomplished in 278 patients, with no major morbidity; 262 of these patients underwent sampling before and after administration of ovine CRH. The adrenocorticotropin levels in the samples were used to calculate the ratio of the concentration in plasma from the inferior petrosal sinuses to the concentration in peripheral-blood plasma (the IPS:P ratio). RESULTS: The diagnosis of 246 patients was confirmed surgically as Cushing's disease in 215, as ectopic adrenocorticotropin syndrome in 20, and as primary adrenal disease in 11. An IPS:P ratio greater than or equal to 2.0 in basal samples identified 205 of the 215 patients with Cushing's disease (sensitivity, 95 percent), with no false positive results (specificity, 100 percent). A peak IPS:P ratio greater than or equal to 3.0 after CRH administration identified all 203 of the patients with Cushing's disease who received CRH (sensitivity, 100 percent), with no false positive results (specificity, 100 percent). The sensitivity was much lower when the adrenocorticotropin concentrations in the samples from one sinus were considered alone. In patients with Cushing's disease a difference of greater than or equal to 1.4-fold between the concentrations in the two sinuses (the adrenocorticotropin gradient) predicted the location of the microadenoma in 68 percent of 104 patients during basal sampling and in 71 percent of 105 patients after CRH administration. CONCLUSIONS: Simultaneous bilateral sampling of plasma from the inferior petrosal sinuses, with the adjunctive use of CRH, distinguishes patients with Cushing's disease from those with ectopic adrenocorticotropin secretion with high diagnostic accuracy.

ACTH Syndrome, Ectopic↗

PubMed1991

Purification and tissue level of the beta-amyloid peptide precursor of rat brain.

The beta-amyloid peptide precursor (beta-APP) exists in brain tissue as a membrane-associated protein extractable with 1% Triton X-100. beta-APP has been purified to near homogeneity by the following procedure: 1) anion exchange chromatography, 2) affinity chromatography on heparin agarose, and 3) immunoaffinity adsorption on matrix-bound antibodies directed to a synthetic peptide corresponding to the last 24 amino acids of the cDNA derived amino acid sequence of beta-APP. Conditions were chosen to minimize denaturation of the protein. The identity of the protein was confirmed by its immunoreactivity with antisera directed to five subsequences derived from the cDNA sequence. The amino-terminal sequence of beta-APP was found to be Leu-Glu-Val-Pro-Thr-Asp-Gly-Asn-Ala-Gly-Leu-Leu-Ala-Glu-Pro, which commences at residue 18 of the cDNA-derived primary structure. The procedure resulted in a 2000-fold purification of beta-APP. The purified protein migrated on polyacrylamide gels as a doublet of apparent molecular mass 100-120 kDa, although the predicted molecular mass of its constituent amino acids is 76 kDa. beta-APP clearly behaves anomalously in gel electrophoresis. The beta-APP content of rat brain amounted to 46 micrograms/g tissue. The half-life of the protein was calculated to be about 10 h, which is 30 times as long as that observed by others in transfected PC-12 cells. We conclude that transfected cell systems may not be adequate models for beta-APP processing.

Amino Acid Sequence↗

PubMed1991

Expression of human Alzheimer amyloid precursor protein in insect cells.

The amyloid beta-peptide is a major constituent of amyloid deposited in the brains of patients with Alzheimer's disease and is derived from a larger precursor protein/s (APP-695, 751, 770). A human cDNA encoding full-length APP-751 was inserted into the genome of Autographa californica nuclear polyhedrosis virus under transcriptional regulation of the viral polyhedrin gene promoter. The recombinant virus was used to infect insect cells, which resulted in the abundant expression of APP-751. Analysis of infected cell proteins indicate that APP-751 is localized in the membrane fraction; however, a significant amount of the protein was cleaved and released into the medium. The NH2-terminal sequence of recombinant APP-751 from the membrane fraction was identical to that of mammalian APP. Immunoblot analysis suggests that the secreted form results from cleavage within the beta-peptide.

Alzheimer Disease↗

PubMed1991

RNA editing by cytidine insertion in mitochondria of Physarum polycephalum.

A corollary of the central dogma of molecular biology is that genetic information passes from DNA to RNA by the continuous synthesis of RNA on a DNA template. The demonstration of RNA editing (the specific insertion, deletion or substitution of residues in RNA to create an RNA with a sequence different from its own template) raised the possibility that in some cases not all of the genetic information for a trait residues in the DNA template. Two different types of RNA editing have been identified in mitochondria: insertional editing represented by the extensive insertion (and occasional deletion) of uridine residues in mitochondrial RNAs of the kinetoplastid protozoa and the substitutional editing represented by the cytidine to uridine substitutions in some plant mitochondria. These editing types have not been shown to be present in the same organism and may have very different mechanisms. RNA editing of both types has been observed in nonmitochondrial systems but is not as extensive and may involve still different mechanisms. Here we report the discovery of extensive insertional RNA editing in mitochondria from an organism other than a kinetoplastid protozoan. The mitochondrial RNA apparently encoding the alpha subunit of ATP synthetase in the acellular slime mould, Physarum polycephalum, is edited at 54 sites by cytidine insertion.

Amino Acid Sequence↗

PubMed1991

Gastric volvulus in the pediatric population.

Acute gastric volvulus in children is uncommon. Since 1899, only 77 cases have been documented in the world literature. In children, mesenteroaxial volvulus is the most common type and associated anatomic defects are the rule. Diagnostic delays result in gastric ischemia, perforation, and death. Nonoperative mortality is 80%. Early recognition, surgical reduction with gastric fixation, and repair of associated defects are the mainstays of therapy for acute gastric volvulus in the pediatric population.

Acute Disease↗

PubMed1991

Evaluation of changes in diatom mobility after exposure to 16-Hz electromagnetic fields.

The effect of a 16-Hz electromagnetic field on the mobility of the diatom Amphora coffeaformis was examined on agar plates that contained no added calcium and also on agar plates containing 0.25 or 2.5 mM exogenous Ca2+. Exposure conditions consisted of an ac field of 16 Hz with an amplitude of 20.9 microT parallel to the horizontal component of the dc field (BH = 20.9 microT, where BV = 0). To assess results, the percentage of diatoms that moved a distance greater than their body length was determined. We observed the field-associated increase in diatom motion at 0.25 mM Ca++, which was previously reported in the literature. Although the magnitude of the effect at 16 Hz was significant, the percentage of cells that moved was not sufficiently reproducible to allow examination for frequency dependence.

Calcium↗

PubMed1991

Miniature-probe measurements of electric fields and currents induced by a 60-Hz magnetic field in rat and human models.

A miniaturized probe was designed and built to provide detailed data on fields induced by a uniform 60-Hz magnetic field in homogeneous models of rat and human. The probe employed three silver wires twisted and potted in an 8-cm hypodermic needle. The exposed tips of the wires formed three sensing electrodes with a centered ground; highly sensitive voltage measurements were enabled by a lock-in amplifier. Tests were conducted in a 1-mT rms field that was uniform within +/- 5%. The models were made by casting 1.5% agar at 1-S/m conductivity into plastic-foam molds. The rat model was scaled 1:1 as an adult (22 cm length; mass about 640 g). The human model was scaled 1:4 as an adult (height = 46.5 cm; mass 1.4 kg). The probe was inserted into each model in several regions, and readings of induced fields were made under different exposure geometries. Maximal strengths of fields induced near the surface of the torso were as high as 120 microV/cm in the laterally exposed rat model. Data extrapolated from the quarter-scale human model revealed that an induced field as high as 700 microV/cm could occur at the torso of a frontally exposed human adult. An overall size-scale factor of about 5 appears to be appropriate for experimental exposures of rats that are intended to simulate currents induced in human beings by magnetic fields. The average strength of electric fields induced in the torso by a 1-mT magnetic field is comparable to that by a vertical electric-field at 60 kV/m and 28 kV/m, respectively, for the rat and human.

Animals↗

PubMed1991

Electric fields induced in chicken eggs by 60-Hz magnetic fields and the dosimetric importance of biological membranes.

Chicken eggs are convenient models for observing the effects of inhomogeneities and variations, such as those found in biological membranes and in cellular conductivities, on the distribution of internal electric fields as induced by exposure to magnetic fields. The vitelline membrane separates the yolk, which has a conductivity of 0.26 S/m, from the white, which has a conductivity of 0.85 S/m. A miniaturized probe with 2.4-mm resolution was used to measure induced fields in eggs placed in a uniform, 1-mT magnetic field at 60 Hz. The E fields induced in eggs with homogenized contents agreed with expectations based on simple theory. Results were similar to intact eggs unless the probe moved the yolk off-center, which greatly perturbed the induced fields. A more reproducible arrangement, which consisted of saline-agar filled dishes with a hole cut for test samples, was developed to enhance definition of electrical parameters. With this test system, the vitelline membrane was found to be responsible for most of the perturbation of the induced field, because it electrically isolates the yolk from the surrounding white. From a theoretical viewpoint, this dosimetry for the macroscopic egg yolk is analogous to the interaction of fields with microscopic cells. These findings may have important implications for research on biological effects of ELF electromagnetic fields, especially for studies of avian embryonic development.

Animals↗

PubMed1991

Localization of parathyroid tumors in patients with asymptomatic hyperparathyroidism and no previous surgery.

A number of recently published series were reviewed evaluating noninvasive localizing studies (ultrasound, thallium-technetium subtraction scintigraphy, computed tomography, and magnetic resonance imaging) in patients with primary hyperparathyroidism and no previous surgery. The average true positive (%) and false positive (%) rates were (1) ultrasound, 66 and 12; (2) Th/Tc scanning 55 and 13; (3) computed tomography, 63 and inadequately documented; (4) magnetic resonance imaging, 75 and 18. The success rate of initial operation in this group of patients is over 90%. There is no evidence that preoperative localizing studies shorten operating time or prevent surgical failures. For these reasons, noninvasive localization studies are not indicated in patients with primary hyperparathyroidism before initial surgery.

Humans↗

PubMed1991

Immunolocalization of Alzheimer beta-amyloid peptide precursor to cellular membranes in baculovirus expression system.

One characteristic of Alzheimer's disease (A beta disease) is the accumulation of amyloid deposits within the extracellular space of the brain and meninges. A 40 amino acid peptide called beta-peptide or A4 protein is the subunit of the amyloid fibrils found in these deposits. The sequence of beta-peptide is contained within those of a family of larger proteins called the Alzheimer beta-amyloid peptide precursor (APP). These APPs contain, in addition to a signal sequence, a hydrophobic sequence that is believed to span cell membranes. Although biochemical studies indicate that some APPs have properties of integral membrane proteins, morphological confirmation of this has not been reported. We recently described an expression system in which human APP751 cDNA was placed under the transcriptional regulation of the polyhedrin gene promoter in the baculovirus Autographica californica infecting a Spodoptera frugiperda cell line (Ramakrishna et al., Biochem Biophys Res Commun 174:983-989, 1991). As part of a larger biochemical and molecular biological study of APP, we have carried out an immunocytochemical study using antibodies directed against several epitopes within APP to reveal, at both the light and the electron microscopic levels, the cellular localization of APP in the baculovirus expression system. These studies demonstrate that APP751 is abundantly synthesized and inserted into certain of the membrane compartments of the cell. As early as 24 hr postinfection, APP751 is found associated with all membrane compartments excepting mitochondrial membranes. The patterns of immunolabeling are consistent with our biochemical findings that the protein is processed in these cells so as to release the extracellular domain and to retain a transmembrane and intracellular segment. These data provide the first morphological demonstration of the membrane location of APP751, its posttranslational processing to a secreted fragment, and its exclusion from the mitochondrial membranes. This system is especially valuable for identifying conditions under which antibodies raised against APP or appropriate synthetic peptides will react with native APP.

Amyloid beta-Protein Precursor↗

PubMed1991

Acute pulmonary emboli associated with guidewire change of a central venous catheter.

The potential complications of percutaneous venous catheterizations are many and include pneumothorax, subclavian and carotid artery puncture, hematoma, air embolism, catheter malposition, catheter fragment embolization, venous thrombosis and infection. This case report describes a patient who developed angiographically documented pulmonary emboli associated with the changing of a central venous catheter over a guidewire using Seldinger technique. This episode adds the possibility of acute pulmonary emboli to the list of potential complications from central venous catheterizations.

Acute Disease↗

PubMed1991

Distribution and activity of alternatively spliced Alzheimer amyloid peptide precursor and scrapie PrP mRNAs on rat brain polysomes.

Mammalian brains contain low levels of the Alzheimer amyloid precursor variants (AAPPs) and the normal form of the scrapie agent protease-resistant protein (PrPc); however, their mRNAs are readily detectable. To understand these discrepancies we have investigated some aspects of the translational regulation of these mRNAs. An accurate blot-hybridization procedure was developed to measure absolute amounts of mRNA. Rat brain contains the following mRNA levels (ng/g tissue) AAPP(695), 170; AAPP(751/770), 63; PrPc, 144; actin, 615; glyceraldehyde-3-phosphate dehydrogenase (G3PDH), 359; ferritin, 148. The method was also used to determine the distribution of mRNAs between translationally active polysomes and translationally inactive ribonucleoprotein protein particles (mRNPs). More than 90% of G3PDH and actin mRNAs were associated with polysomal RNA; whereas, ferritin light chain mRNA was predominantly (90%) in mRNP RNA. The degree of cross-contamination of mRNPs with polysomes was less than 10%. Probes specific for the scrapie PrP protein and the AAPP(695) splice junction revealed that 70% of these mRNAs were associated with polysomes. One-half of AAPP(751/770) mRNAs (which comprise 20-30% of all AAPP mRNA in brain) were found in polysomes. We conclude therefore that both scrapie and AAPP mRNAs are subject to translational regulation in rat brain. Evidence from in vitro translational experiments confirm the message distribution determined by blot hybridization and corroborate the hypothesis that AAPP is subject to partial post-transcriptional regulation. Nevertheless, the low tissue levels of AAPP and PrPc must result primarily from their relatively rapid turnover.

Amyloid beta-Peptides↗

PubMed1991

Nucleotide and amino acid sequence variation in the L1 and E7 open reading frames of human papillomavirus type 6 and type 16.

Human papillomavirus (HPV) type 6 and type 16 DNA sequence variants were found by partially sequencing the L1 and E7 open reading frames, using templates generated with the polymerase chain reaction. Identical variants were found in patients from widely separated locations, such as the United States, the Philippines, and India. The same sequence variants of HPV 16 were found in women with invasive cervical carcinoma and in women with no evidence of disease. Variation in the predicted amino acid sequences of the HPV 16 L1 and E7 proteins was found. A single nucleotide change at position 6433 was found in about 50% of the HPV 16 DNAs, resulting in a change in predicted amino acid sequence from threonine to alanine at the equivalent position in the L1 protein. Predicted amino acid changes were found in the HPV 16 E7 proteins at amino acid positions 28, 29, and 47. Variation at these positions could affect known properties of the E7 protein, including binding to the retinoblastoma protein.

Amino Acid Sequence↗

PubMed1991

Ultrasonic cavitation indirectly induces single strand breaks in DNA of viable cells in vitro by the action of residual hydrogen peroxide.

Direct exposure of cells to vigorous ultrasonic cavitation results predominantly in mechanical cell lysis, but latent effects due to production of toxic sonochemicals can also be present. Phosphate buffered saline (PBS) was exposed to 1.61 MHz ultrasonic cavitation at 20 degrees C in a rotating tube exposure system to build up sonochemical products. Single strand DNA breaks (SSBs) were then induced by treating Chinese hamster ovary (CHO) cells with the cavitated PBS for 30 min on ice. The SSBs resided in viable cells, as evidenced by their ability to repair the breaks when warmed. This indirect effect could be explained by the action of cavitation-generated hydrogen peroxide that had built up (e.g., to 16 microM after 30 min exposure) in the PBS. Dissolution of argon gas in the PBS before exposure enhanced the SSB effect and the H2O2 production. Addition of catalase to the cavitated PBS before cell treatment eliminated the H2O2 and the SSB gamma effect. Tests with hydrogen peroxide showed that 16 microM H2O2 treatment for 30 on ice was as effective as 1 Gy dose of 60Co gamma rays in producing single strand breaks. The SSB effect of H2O2 and gamma rays was reduced by addition of the radical scavenger cysteamine to the cells before treatment, but cysteamine did not reduce the SSB effect of direct exposure to ultrasonic cavitation. These results help to clarify the potential for genetic effects from ultrasonic cavitation. These effects help to clarify the potential for genetic effects from ultrasonic cavitation.

Animals↗

PubMed1991

Mechanisms for hemolysis by ultrasonic cavitation in the rotating exposure system.

The rotating chamber provides a useful system for enhancing cavitation bioeffects so that they may be more easily studied. A tube with acoustically transparent windows was rotated at 72 rpm perpendicular to its axis. The 1.61 MHz ultrasound beam was switched on and off in 42 ms bursts to expose the chamber while it was aligned with the beam. The hemolysis of 0.5% suspensions of canine erythrocytes was used as a measure of the efficacy of this rotating exposure system. Use of dialysis membrane, agar plugs, plastic sealing film or wax film for the windows produced no differences in the results for exposure intensities above an apparent threshold of 2-2.8 W/cm2, up to 16 W/cm2, at which essentially 100% lysis was observed. Changing the tube length from 1.4 cm to 4 cm, or partitioning it into three 0.5 cm segments had little influence on the results, but a short 0.5 cm chamber had reduced efficacy. Pressurizing the suspension filled chamber at 10 MPa to reduce the population of cavitation nuclei reduced the hemolysis. Separately pressurizing the suspension or the chamber produced indistinguishable, smaller reductions in hemolysis. The results lead to the hypothesis that the hemolysis occurs primarily in the bulk of the medium (rather than on surfaces). Bubbles recycled by rotation into the medium move toward the back of the chamber at estimated speeds of 10 m/s, generating hydrodynamic stresses sufficient to cause the observed hemolysis.

Acoustics↗

PubMed1991

Single strand breaks in CHO cell DNA induced by ultrasonic cavitation in vitro.

Ultrasonic cavitation induces a multiplicity of bioeffects in cell suspensions exposed in a 72 RPM rotating-tube exposure system. Single strand DNA breaks (SSBs) were found in cultured Chinese hamster ovary (CHO) cells exposed directly to 1.61 MHz ultrasound at a continuous 8 W/cm2 spatial peak temporal average (SPTA) intensity with cavitation for 10 min at 2 degrees C. Viability assessed by the trypan blue test was less than 1%, which indicates that these SSBs were in dead cells. Burst mode exposure with 10.5 microseconds bursts repeated each 21 microseconds not only caused SSBs at 5.6 W/cm2 and 8 W/cm2 (SPTA) for 10 min, but also allowed 20% and 7% viability, respectively. In order to determine if any of these breaks resided in the viable fraction of cells, the exposures were repeated with a 30 min postexposure incubation period at 37 degrees C to allow breaks in viable cells to repair. No significant repair occurred, relative to the samples which remained at 2 degrees C to prevent repair. A similar result was obtained with 10.5 microseconds bursts repeated each 42 microseconds at 4 W/cm2 (SPTA) with 46% viability. Thus, the observed ultrasonically induced SSBs reside primarily in the nonviable fraction of cells.

Animals↗

PubMed1991

Selective catheterization of the inferior petrosal sinuses: new catheter design.

Catheters for selective catheterization of the right and left inferior petrosal sinuses have been developed to replace the complex tip-deflector catheter-guide-wire system currently used. The new catheters are easily formed from commonly available straight catheters with the use of steam. They have been successfully tested in 22 patients; the only complications were minor groin hematomas.

ACTH Syndrome, Ectopic↗

PubMed1991

Petrosal sinus sampling: technique and rationale.

Bilateral simultaneous sampling of the inferior petrosal sinuses is an extremely sensitive, specific, and accurate test for diagnosing Cushing disease and distinguishing between that entity and the ectopic ACTH syndrome. It is also valuable for lateralizing small hormone-producing adenomas within the pituitary gland. The inferior petrosal sinuses connect the cavernous sinuses with the ipsilateral internal jugular veins. The anatomy of the anastomoses between the inferior petrosal sinus, the internal jugular vein, and the venous plexuses at the base of the skull varies, but it is almost always possible to catheterize the inferior petrosal sinus. In addition, variations in size and anatomy are often present between the two inferior petrosal sinuses in a patient. Advance preparation is required for petrosal sinus sampling. Teamwork is a critical element, and each member of the staff should know what he or she will be doing during the procedure. The samples must be properly labeled, processed, and stored. Specific needles, guide wires, and catheters are recommended for this procedure. The procedure is performed with specific attention to the three areas of potential technical difficulty: catheterization of the common femoral veins, crossing the valve at the base of the left internal jugular vein, and selective catheterization of the inferior petrosal sinuses. There are specific methods for dealing with each of these areas. The sine qua non of correct catheter position in the inferior petrosal sinus is demonstration of reflux of contrast material into the ipsilateral cavernous sinus. Images must always be obtained to document correct catheter position. Special attention must be paid to two points to prevent potential complications: The patient must be given an adequate dose of heparin, and injection of contrast material into the inferior petrosal sinuses and surrounding veins must be done gently and carefully. When the procedure is performed as outlined, both inferior petrosal sinuses can be catheterized in more than 98% of patients. The complication rate is low, and the theoretical risk of major morbidity or death is less than 1% (neither has yet occurred, to our knowledge). The most common complication is groin hematoma.

ACTH Syndrome, Ectopic↗