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Biomedical subjects

K A Davis

Publications and source records attributed to K A Davis.

At least 19 recordsLinked to original sources

Use of a high affinity DNA ligand in flow cytometry.

To investigate the feasibility of using oligonucleotides in flow cytometry we describe a model system consisting of human neutrophil elastase (HNE) coated on 3.3 micro beads and a high affinity DNA ligand for HNE isolated by in vitro selection (SELEX). In this system the fluoresceinated DNA ligand was equally effective as an anti- HNE antibody in detecting HNE on beads. The location on and the chemistry of attachment of fluorescein to the DNA ligand is critical for the sensitivity of detection. DNA constructs in which fluorescein was conjugated via an ethylene glycol tether to either the 5'-end or near the 3'-end gave much higher signals than did probes with fluorescein directly conjugated to either end. Second-step staining with strepavidin-conjugated phycoerythrin was accomplished using a biotinylated DNA ligand in the initial staining of HNE beads. These data suggest that instead of, or in addition to, antibodies high affinity oligonucleotide probes can be useful in diagnostic applications based on flow cytometry.

Animals

Computed tomography in blunt hepatic trauma.

BACKGROUND: Nonoperative management of blunt hepatic injury in hemodynamically stable trauma patients is now common. Recently, it has been proposed that the finding of hepatic periportal tracking (PPT) of blood on the initial computed tomographic (CT) scan is a sensitive marker of significant hepatic and subhepatic injury that might militate against nonoperative management. While CT scan is useful in diagnosing the injury, the utility of follow-up CT scans has not been elucidated. DESIGN: Retrospective chart review. SETTING: Regional trauma center. PATIENTS: The records of 58 hemodynamically stable patients with blunt hepatic trauma were reviewed and the following data recorded: age, outcome, Injury Severity Score (ISS), operative intervention, and complications. Computed tomographic scans were taken on admission and reviewed for the presence of PPT. The timing and radiographic appearance of follow-up CT scans was also recorded. RESULTS: Seventeen patients (29%) had radiographic evidence of PPT while 41 patients (71%) did not. Age, ISS, injury grade, overall success rate of nonoperative management, and incidence of complications were not statistically significant between the two groups. In no instance did a routine follow-up CT scan affect subsequent management of the patient. CONCLUSIONS: The finding of PPT on the admission CT scan is not clinically significant and does not preclude nonoperative management of patients with blunt hepatic injury. Furthermore, routine follow-up CT scans are not indicated, as treatment is not influenced by their results. Rather, follow-up CT scans should be obtained as dictated by the patient's clinical course. Extrapolation of these findings to all patients with blunt hepatic trauma in the United States would result in considerable savings of health care dollars, without negatively affecting patient care.

Adolescent

Response properties of units in the dorsal cochlear nucleus of unanesthetized decerebrate gerbil.

1. The electrophysiological responses of single units in the dorsal cochlear nucleus of unanesthetized decerebrate Mongolian gerbil (Meriones unguiculatus) were recorded. Units were classified according to the response map scheme of Evans and Nelson as modified by Young and Brownell, Young and Voigt, and Shofner and Young. Type II units have a V-shaped excitatory response map similar to typical auditory nerve tuning curves but little or no spontaneous activity (SpAc < 2.5 spikes/s) and little or no response to noise. Type I/III units also have a V-shaped excitatory map and SpAc < 2.5 spikes/s, but have an excitatory response to noise. Type III units have a V-shaped excitatory map with inhibitory sidebands, SpAc > 2.5 spikes/s, and an excitatory response to noise. Type IV-T units typically also have a V-shaped excitatory map with inhibitory sidebands, but have a highly nonmonotonic rate versus level response to best frequency (BF) tones like type IV units, SpAc > 2.5 spikes/s, and an excitatory response to noise. Type IV units have a predominantly inhibitory response map above an island of excitation of BF, SpAc > 2.5 spikes/s, and an excitatory response to noise. We present results for 133 units recorded with glass micropipette electrodes. The purpose of this study was to establish a normative response map data base in this species for ongoing structure/function and correlation studies. 2. The major types of units (type II, type I/III, type III, type IV-T, and type IV) found in decerebrate cat are found in decerebrate gerbil. However, the percentage of type II (7.5%) and type IV (11.3%) units encountered are smaller and the percentage of type III (62.4%) units is larger in decerebrate gerbil than in decerebrate cat. In comparison, Shofner and Young found 18.5% type II units, 30.6% type IV units, and 23.1% type III units using metal electrodes. 3. Two new unit subtypes are described in gerbil: type III-i and type IV-i units. Type III-i units are similar to type III units except that type III-i units are inhibited by low levels of noise and excited by high levels of noise whereas type III units have strictly excitatory responses to noise. Type IV-i units are similar to type IV units except that type IV-i units are excited by low levels of noise and become inhibited by high levels of noise whereas type IV units have strictly excitatory responses to noise. Type III-i units are approximately 30% of the type III population and type IV-i units are approximately 50% of the type IV population. 4. On the basis of the paucity of classic type II units and the reciprocal responses to broadband noise of type III-i and type IV-i units, we postulate that some gerbil type III-i units are the same cell type and have similar synaptic connections as cat type II units. 5. Type II and type I/III units are distinguished from one another on the basis of both their relative noise response, rho, and the normalized slope of the BF tone rate versus level functions beyond the first maximum. Previously, type II units were defined to be those nonspontaneously active units with rho values < 0.3 where rho is defined as the ratio of the maximum noise response minus spontaneous rate to the maximum BF tone response minus spontaneous rate. In the gerbil, the average rho value for type II units is 0.25, although a few values are > 0.3, and the rate-level curves are consistently nonmonotonic with normalized slopes steeper than than -0.007/dB. The average rho value for type I/III units is 0.54, although a few values are < 0.3, and the rate-level curves tend to saturate with slopes shallower than -0.006/dB. In general, the response properties of type II units recorded in gerbil are similar to those recorded in decerebrate cat. 6. In comparison to decerebrate cat, the lower percentage of type IV units recorded in decerebrate gerbil may be due to a species difference (a reduced number of type II units in gerbil) or an electrode bias.

Animals

A statistically based method to generate response maps objectively.

One scheme to classify the physiological response properties of single units in the cochlear nucleus is based on the average discharge rate of the unit and is reflected in the distribution of excitatory and inhibitory regions in a frequency-level map (response map) that spans the unit's receptive area (e.g., Evans and Nelson, 1973; Young and Brownell, 1976; Young and Voigt, 1982; Shofner and Young, 1985, Spirou and Young, 1991). Typically, discharge rate versus level curves are acquired at many frequencies and the investigator determines that a unit is excited or inhibited at a given level if the driven rate is above or below a spontaneous rate estimate by a specified criterion (for example, 20%). The investigator then encloses regions of excitation and inhibition where responses over adjacent frequencies and levels are consistent. In the present report, we describe an objective 3-step computer-based method to generate response maps: raw driven and spontaneous rate estimates are smoothed with a low-pass spatial filter; a unit is said to be excited or inhibited at a given level if the filtered driven rate is above or below the mean filtered spontaneous rate for that frequency by a specified criterion (percentage or statistical); and resultant response maps are median spatial filtered to eliminate spurious regions. The results shown here demonstrate that use of a statistical criterion provides a more reliable detection of excitation and inhibition than a 20% criterion, particularly when the variance of the rate estimates is high. Further, the statistically based method permits unit classification based on response map data that are more rapidly acquired with shorter duration stimuli (32 vs. 200 ms). Although this method is applied to units recorded in the dorsal cochlear nucleus, the technique may be applicable to studies of receptive fields and their plasticity in other systems.

Acoustic Stimulation

Neural modeling of the dorsal cochlear nucleus: cross-correlation analysis of short-duration tone-burst responses.

A conceptual model of a portion of dorsal cochlear nucleus (DCN) neural circuitry has emerged over the past two decades. This model suggests that the response properties of the DCN's major projection neurons, called type IV units, are due, in part, to the behavior of local circuit inhibitory interneurons called type II units (Young and Brownell 1976). Cross-correlation studies of simultaneously recorded pairs of DCN units in decerebrate cat derived from 50-s best frequency (BF) stimuli are consistent with and have extended this conceptual model (Voigt and Young 1980, 1985, 1988, 1990). Interestingly, Gochin et al. (1989) found no signs of inhibition in the anesthetized rat DCN in cross-correlograms derived from 55-ms short-duration BF tone bursts. This seemingly contradictory result has motivated this study. Computer simulations were run using our network model of the intrinsic DCN neural circuitry. This model has previously been shown to reproduce the major features of both type II and type IV rate-level curves and the inhibitory trough (IT) observed in cross-correlograms derived from long-duration stimuli (Voigt and Davis 1994). The goal was to study the stimulus-duration-dependent strength of ITs in the cross-correlograms derived from short-duration BF tone-burst stimuli. The results suggest that ITs may not be detectable when the stimulus duration is 50 ms but may be detectable when the stimulus duration is 200 ms or greater. Furthermore, when the ITs are detected in cross-correlograms derived from 200-ms data sets, the strength of the IT, as measured by effectiveness, is comparable to the strength of ITs measured when the stimulus duration is 50 s.

Acoustic Stimulation

Malignant degeneration of pilonidal cysts.

Malignant degeneration is a rare occurrence in pilonidal disease. The authors present three new cases with a review of the world's previously published 41 cases. Among the total 44 cases, 36 were squamous cell carcinoma. All cases occurred in the setting of long-standing pilonidal disease, with the mean duration of antecedent disease being 23 years. Five of six patients presenting with inguinal metastases died within 16 months. Four patients received adjuvant radiation therapy, one received adjuvant chemotherapy, and one patient in the current series received both adjuvant chemotherapy and radiation. Six patients with recurrence underwent potentially curative resection, with three patients surviving greater than 10 years with no evidence of disease. The authors propose consideration of adjuvant chemotherapy and radiation as a new modality to decrease the local recurrence rate.

Adult

Respiratory impedance spectral estimation for digitally created random noise.

Measurement of respiratory input mechanical impedance (Zrs) is noninvasive, requires minimal subject cooperation, and contains information related to mechanical lung function. A common approach to measure Zrs is to apply random noise pressure signals at the airway opening, measure the resulting flow variations, and then estimate Zrs using Fast-Fourier Transform (FFT) techniques. The goal of this study was to quantify how several signal processing issues affect the quality of a Zrs spectral estimate when the input pressure sequence is created digitally. Random noise driven pressure and flow time domain data were simulated for three models, which permitted predictions of Zrs characteristics previously reported from 0-4, 4-32, and 4-200 Hz. Then, the quality of the Zrs estimate was evaluated as a function of the number of runs ensemble averaged, windowing, flow signal-to-noise ratio (SNR), and pressure spectral magnitude shape magnitude of P(j omega). For a magnitude of P(j omega) with uniform power distribution and a SNR less than 100, the 0-4 Hz and 4-200 Hz Zrs estimates for 10 runs were poor (minimum coherence gamma 2 less than 0.75) particularly where Zrs is high. When the SNR greater than 200 and 10 runs were averaged, the minimum gamma 2 greater than 0.95. However, when magnitude of P(j omega) was matched to magnitude of Zrs, gamma 2 greater than 0.91 even for 5 runs and a SNR of 20. For data created digitally with equally spaced spectral content, the rectangular window was superior to the Hanning. Finally, coherence alone may not be a reliable measure of Zrs quality because coherence is only an estimate itself. We conclude that an accurate estimate of Zrs is best obtained by matching magnitude of P(j omega) to magnitude of Zin (subject and speaker) and using rectangular windowing.

Airway Resistance

Continuous liquid-phase piezoelectric biosensor for kinetic immunoassays.

The reactions of immunoglobulins with protein A and, subsequently, of antibodies to these immunoglobulins were continuously monitored by a piezoelectric biosensor. AT-cut crystals, with a fundamental resonant frequency of 10 MHz, were mounted in a Plexiglas holder, and one side was directly coated with protein A. Upon exposure to solutions containing rabbit or human IgG the resonant frequency was followed continuously and its decrease due to the binding of IgG to protein A observed. Subsequent addition of sheep antihuman IgG to the now immobilized human IgG caused a specific 3-fold further decrease in resonant frequency. We observed a frequency change of approximately 1 Hz fore each 10 ng of added immunoglobulin. Decreasing the pH to 3 released the bound IgG but not the protein A and permitted reuse of the crystal for further IgG binding.

Biosensing Techniques

Effects of hyperoxia and dimethylthiourea on neonatal pulmonary hemodynamics in piglets.

Exposure of piglets to 68-72 h of hyperoxia has previously been shown to blunt hypoxic pulmonary vasoconstriction (HPV). We tested the hypothesis that the administration of a scavenger of toxic oxygen radicals during exposure to hyperoxia would preserve HPV in piglets. Newborn piglets were kept in FIO2 greater than 0.90 for 68-72 h and compared to control animals kept in room air. Randomly selected animals from both groups were given the O2 metabolite scavenger, dimethylthiourea (DMTU) (500 mg/kg followed by 250 mg/kg/d), resulting in plasma levels of 2-6 mM. Following the oxygen or room air (RA) exposure period, piglets had pulmonary vascular resistance (PVR) measured following a 20 min exposure to alveolar hypoxia (FIO2 = 0.12). Both groups of RA-exposed animals (DMTU and saline treatment) as well as the hyperoxia-exposed saline group had almost 2-fold increases in PVR during exposure to hypoxia (P less than 0.05). Contrary to expectations, the PVR in the hyperoxia-exposed DMTU group did not rise significantly during hypoxia and the use of DMTU did not restore HPV by increasing PVR to levels greater than the hyperoxia/saline group (P = 0.70). Therefore DMTU does not appear to prevent hyperoxia-induced blunting of HPV in piglets.

Animals

White matter lesions: role of spin density in MR imaging.

To study the effect of hydrogen spin density (N[H]) on magnetic resonance (MR) image contrast in white matter disease of the brain, T1, T2, and N[H] values were determined for normal white matter and idiopathic white matter lesions (IWMLs) in 21 patients by using multiple spin-echo (SE) sequences. T1 values of IWMLs were significantly greater than those of normal white matter in all patients studied, and T2 values of IWMLs were significantly greater in 20 of 21 patients. N[H] values of IWMLs were greater in 20 of 21 patients, with statistically significant differences from those of normal white matter in 17 of those 20 patients. Averaged over all 21 patients studied, N[H] values of IWMLs were 20% higher than N[H] values of normal white matter. The effect of unequal N[H] values on contrast between IWMLs and normal tissues is to reduce contrast on short SE sequences with a short repetition time (TR) and a short echo time (TE), while enhancing contrast between IWMLs and normal tissues on long TR/TE SE sequences. Elevated N[H] values in IWMLs have a minimal effect on contrast in conventional inversion-recovery (IR) sequences but substantially enhance contrast between IWMLs and normal brain tissues in short inversion time IR sequences.

Brain

Toluene abuse causes diffuse central nervous system white matter changes.

We describe the findings of magnetic resonance imaging (MRI) of the brain in 6 chronic toluene vapor abusers and the neuropathological findings in 1 abuser not studied by MRI. MRI in 6 chronic toluene abusers revealed the following abnormalities: (1) diffuse cerebral, cerebellar, and brainstem atrophy; (2) loss of differentiation between the gray and white matter throughout the central nervous system; and (3) increased periventricular white matter signal intensity on T2-weighted images. Another chronic toluene abuser (MRI not performed) died as a result of acute toluene overdose. The brain displayed diffuse, ill-defined myelin pallor, maximal in cerebellar, periventricular, and deep cerebral white matter. Neurons were preserved throughout, axonal swelling or beading was not seen, gliosis was minimal, and occasional, scant perivascular macrophage collections were seen. Taken in concert, these findings suggest that the pathological and MRI abnormalities are due to either increased water content of the white matter or subtle toluene-induced metabolic changes in myelin.

Brain

Ascorbic acid status: biochemical and clinical considerations.

Ascorbic acid (AA) plus dehydroascorbic acid (DHAA) was stable in whole blood in unopened vacutainer tubes. Oxidation of DHAA in plasma was irreversible and rapid at 25 degrees C but not at 4 degrees C. The half-life of AA + DHAA in total parenteral nutrition solutions was greater than 24 h. Conditions are described for dithiothreitol reduction of DHAA followed by assay of total AA, with DHAA being determined by difference. DHAA accounted for less than 5% of the total plasma AA + DHAA in both healthy and diabetic subjects. Infection and postpartum-induced stresses resulted in no significant change in the AA content of polymorphonuclear leukocytes (PMN) per unit of blood but AA content per cell was decreased. When glucose concentration was increased, the Km for AA uptake by PMNs doubled. These chemical and physiological factors combined with the rapid movement of the vitamin between plasma and cells suggest that a reliable indicator of AA status remains to be identified.

Ascorbic Acid

Thrombosed arteriovenous malformations: a type of occult vascular malformation. Magnetic resonance imaging and histopathological correlations.

Thrombosed arteriovenous malformations (AVMs) are the predominant type of occult vascular malformation and do not seem to differ significantly in clinical or radiographic presentation from other types of occult vascular malformations. Thrombosed AVMs and occult vascular malformations occasionally present with symptoms secondary to subacute or occult hemorrhage that requires operation. The histopathology of thrombosed AVMs and occult vascular malformations does not seem to have prognostic significance. Five patients with histologically verified thrombosed AVMs are reported. Two patients had previous incomplete resection of histologically proven thrombosed AVMs and presented with recurrence and bleeding. Two patients presented with seizures and headaches, and one patient presented with hemiparesis and headache. All histology specimens had evidence of hemorrhage. The pathological findings of these lesions seem variable; two specimens contained a mixture of cavernous angioma and AVM. Two previously resected lesions had been defined histologically as thrombosed AVMs. Successful excision of the lesions was accomplished in four patients, and one patient had stereotactic biopsy. The magnetic resonance imaging characteristics of lesions seem to add a degree of specificity over computed tomography and angiography. We have found that the literature poorly describes the natural history and histology of these lesions. Conservative management for seizures and headaches due to thrombosed AVMs may not be warranted because of a propensity for hemorrhage and recurrence.

Adult

Cytochrome a3 deficiency in human achondroplasia.

Mitochondria prepared from tissue culture cells (skin fibroblasts) from normal subjects and subjects with homozygous achondroplasia were studied to determine the concentrations of cytochromes a and a3 in the preparations. Cytochrome a3 was markedly decreased (80%) in the achondroplastic preparations with cytochrome a present in normal amounts. Determination of total heme a (as the pyridine hemochromogen) in the normal and achondroplastic preparations demonstrated that the observed decrease in concentration of cytochrome a3 in the achondroplastic preparations was due to an absence of cytochrome a3 and not to a change in its absorbancy (extinction coefficient). The decreased concentrations of cytochrome a3 in the achondroplastic cells may decrease the reactivity or affinity of the mitochondrial oxidative systems for oxygen and result in the phenotypic expression of the disease.

Achondroplasia

Studies of human achondroplasia: oxidative metabolism in tissue culture cells.

Mitochondria prepared from the first growth of cells (fibroblasts) from skin biopsies from homozygous (but not heterozygous) achondroplastic human subjects were unable to carry out oxidative phosphorylation. However, successive crops of cells gained the ability to phosphorylate with normal P:O ratios with pyruvate-malate and succinate as substrates. Concentrations of cytochromes a + a3 were markedly and significantly lower in homogenates of homozygous achondroplastic tissue culture cells than in homogenates of normal cells. Levels of cytochromes a + a3 in the heterozygous achondroplastic cells were intermediate between the levels in normal cells and the homozygous achondroplastic cells. Activities of the mitochondrial oxidative systems (NADH, succinic and cytochrome oxidases) were not significantly lower in the achondroplastic cell preparations than in normal cell preparations under standard assay conditions (saturation levels of oxygen).

Achondroplasia

Clinical applications of magnetic resonance imaging--current status.

Magnetic resonance imaging has far-reaching real and possible clinical applications. Its usefulness has been best explored and realized in the central nervous system, especially the posterior fossa and brain stem, where most abnormalities are better identified than with computed tomography. Its lack of ionizing radiation and extreme sensitivity to normal and abnormal patterns of myelination make magnetic resonance imaging advantageous for diagnosing many neonatal and pediatric abnormalities. New, reliable cardiac gating techniques open the way for promising studies of cardiac anatomy and function. The ability to image directly in three orthogonal planes gives us new insight into staging and follow-up of pelvic tumors and other pelvic abnormalities. Exquisite soft tissue contrast, far above that attainable by other imaging modalities, has made possible the early diagnosis of traumatic ligamentous knee injury, avascular necrosis of the hip and diagnosis, treatment planning and follow-up of musculoskeletal neoplasms.

Bone Diseases

Studies of pyridine nucleotide oxidizing enzymes from human neutrophils.

An NADH cytochrome c reductase has been identified in plasma membrane fractions from neutrophils in addition to the superoxide producing NADPH oxidase which has been extensively studied by other investigators. Activation of neutrophils resulted in increased enzyme activities but to different degrees; the NADH cytochrome c reductase increased 2 fold in specific activity and the NADPH oxidase 30 fold. Treatment of the plasma membrane fraction with sonication and differential centrifugation yielded a particulate fraction (R2) with a 2 fold increase in specific activities of both enzymes and concentrations of cytochrome b and FAD. The cytochrome b in the preparation was not reduced under anaerobic conditions by either NADH or NADPH. Treatment of preparations of R2 with deoxycholate or potassium thiocyanate separated the two enzymes yielding particulate preparations with only NADPH oxidase or NADH cytochrome c reductase activity, respectively.

Cell Membrane