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Biomedical subjects

D E Lewis

Publications and source records attributed to D E Lewis.

At least 55 records · Page 3Linked to original sources
PubMed1997

Early spontaneous abortions and fetal thymic abnormalities in maternal-to-fetal HIV infection.

The thymus is thought to play a major role in the immunopathogenesis of human immunodeficiency virus (HIV) infection, particularly in maternal-to-fetal HIV transmission. Characteristic lesions of the HIV-infected thymus include a prominent CD4+ CD8+ T lymphocyte depletion at the corticomedullary junction, the region of the thymus where immune selection occurs. At least threefold excess early spontaneous abortions were noted in a cohort of 124 HIV-infected pregnant women. In these 13 abortuses a very high rate (54%) of HIV vertical transmission was documented, with the thymus gland particularly affected. It is possible that the thymic insult in HIV-infected fetuses contributes to immune rejection of the fetus, possibly by an imbalance of maternal and fetal T1- and T2-type cytokines, known to be important in HIV disease progression. We propose, therefore, that the early spontaneous abortions occurring in HIV-infected pregnant women are due, at least in part, to abnormal immune forces created by HIV infection of the thymus.

Abortion, Spontaneous↗
PubMed1997

Granuloma cytokines in murine cysticercosis.

Neurocysticercosis, caused by Taenia solium, is one of the most common causes of seizures worldwide. The symptoms result from granulomatous inflammation associated with dying cyst forms of the parasite. Although the invasive larvae can be killed by immune serum plus complement, immunity to the cyst stage depends on a cellular response. This dichotomous immune response is reminiscent of the extremes of the immune response associated with T helper 1 (Th1) and Th2 cytokine profiles. To characterize the cytokine response in cysticercosis, granulomas were removed from the peritoneal cavity of mice infected with Taenia crassiceps cysts and examined for cytokine message by in situ hybridization using 35S-labeled RNA probes. The granulomas were staged based on histologic appearance of the degenerating parasite. Message for gamma interferon (IFN-gamma) was identified by light microscopy in 11 of the 12 granulomas, and interleukin-2 (IL-2) message was identified in 9 of the 12. By laser scanning confocal microscopy, significantly increased IFN-gamma and IL-2 pixel intensity was identified in nearly all of the granulomas from early histologic stages. Message for IL-4 was seen in 6 of the 12 granulomas. Only granulomas with complete destruction of the parasite architecture displayed more than minimal amounts of IL-4 message by light microscopy, and only 2 of 12 granulomas had IL-4 pixel intensity significantly above background. Only minimal amounts of IL-10 message were detected in 4 of 11 granulomas. Thus, early granulomas in cysticercosis are predominantly associated with a Th1 response, whereas later granulomas, in which parasite destruction is complete, have a mixture of Th1 and IL-4. The Th1 response appears to play an important role both in the pathogenesis of disease as well as in the clearing of the parasites, with IL-4 involved in downregulation of the initial response.

Animals↗
PubMed1996

In vivo and in vitro studies of TrpR-DNA interactions.

The binding of tryptophan repressor (TrpR) to its operators was examined quantitatively using in vitro and in vivo methods. DNA sequence requirements for 1:1 and tandem 2:1 (TrpR:DNA) binding in various sequence contexts were studied. The results indicate that the optimal half-site sequence for recognition by one helix-turn-helix motif of one TrpR dimer is 3'CNTGA5'5'GNACT3', consistent with contacts observed by X-ray diffraction analysis of cocrystalline 1:1 and 2:1 complexes. Half-sites can be paired to form a palindrome either by direct abutment, forming the nucleation site for a tandem 2:1 complex, or with an 8-base-pair spacer, forming a 1:1 target. Dimethylsulfate (DMS) methylation-protection footprinting in vitro of 1:1 and 2:1 complexes formed sequentially on the two unequal half-site pairs of the trpEDCBA operator from Serratia marcescens indicated an obligate hierarchy of site occupancy, with one half-site pair serving as the nucleation site for tandem binding. DMS footprinting of Escherichia coli operators in vivo showed that, over a wide range of intracellular TrpR concentration, the trpEDCBA operator is occupied by three repressor dimers, aroH is occupied by two dimers, and the 1:1 binding mode is used on the trpR operator. The coexistence of these distinct occupancy states implies that changes in protein concentration affect only the fractional occupancy of each operator rather than the binding mode, which is determined by the number of half-site sequences present in the operator region. Cooperativity of tandem complex formation measured by gel retardation using a symmetrized synthetic operator containing identical, optimal sites spaced as in natural operators was found to be modest, implying a maximum coupling free energy of approximately -2 kcal/mol. On other sequences the apparent degree of cooperativity, as well as the apparent affinity, varies with sequence and sequence context in a manner consistent with the structural models and which suggests compensation between affinity and cooperativity as a mechanism that allows tolerance of operator sequence variation.

Bacterial Proteins↗
PubMed1996

Rare event selection of fetal nucleated erythrocytes in maternal blood by flow cytometry.

A noninvasive method of prenatal genetic diagnosis requires fetal cell selection from the maternal circulation that allows efficient recovery for analysis by fluorescence in situ hybridization (FISH). We have solved several problems that negatively affect the isolation and FISH analysis of fetal nucleated red blood cells (nRBCs) in the maternal circulation. The use of glycophorin A (Gly A) antibodies (Abs) for selection is problematic because all five monoclonal antibodies (mAbs) tested caused agglutination of non-nRBCs, thereby changing both light scatter and fluorescence properties of cells by flow cytometry. Because the number of non-nRBCs is variable after Ficoll separation, isolation of nRBCs could be compromised severely by agglutination of nucleated cells with nonnucleated cells, causing them to shift light scatter and fluorescence properties. Several methods for the removal of unwanted maternal white blood cells with CD45 mAbs were also evaluated. Magnetic bead depletion was found to interfere with FISH detection because of residual bead debris after sorting. By contrast, removal of CD45+ cells by a panning technique eliminated this problem. Positive selection methods based on CD71, CD45, and LDS-751 staining and detection of fetal cells by gamma globin expression were also analyzed. Fetal cells were detected by FISH in 11 of 19 (CD71 selection) and in 13 of 15 (gamma selection) random pregnancies. These data support the possibility of a noninvasive method for isolation and analysis of fetal cells for prenatal diagnosis.

Antigens, CD↗
PubMed1996

Effect of hepatocyte proliferation and cellular DNA synthesis on hepatitis B virus replication.

BACKGROUND & AIMS: Interrelationship between hepatitis B virus (HBV) replication and the stage of hepatocyte proliferation and differentiation may play an important role in the pathogenesis of HBV infection. The aim of this study was to assess the effect of hepatocyte proliferation and/or cell arrest on HBV replication. METHODS: Hepatoblastoma cells transfected with HBV were subjected to serum deprivation or treatment with aphidicolin or camptothecin. Cell cycle analysis was performed using flow cytometry, and cellular DNA synthesis was analysed by assessing 5-bromo-2'-deoxyuridine incorporation. Distribution of episomal HBV DNA and proliferating cell nuclear antigen in liver specimens was assessed by simultaneous in situ hybridization and immunohistochemistry. RESULTS: Serum deprivation inhibited cellular DNA synthesis and increased levels of HBV messenger RNA (mRNA). Aphidicolin treatment resulted in cell arrest in C1, with concomitant increases in levels of HBV mRNA and viral DNA. Cell entry into S phase inhibited expression of HBV mRNA. Camptothecin induced G2 cell arrest and inhibited cellular DNA synthesis with increased amounts of viral replication and levels of HBV mRNA. In vivo studies showed an inverse correlation between expression of proliferating cell nuclear antigen and presence of episomal HBV DNA in individual hepatocytes. CONCLUSIONS: HBV replication is cell cycle dependent, supporting the concept of enhanced viral replication in quiescent hepatocytes. The results may explain the mechanism of viral elimination during cell regeneration.

Cell Cycle↗
PubMed1996

Use of bed rest and head-down tilt to simulate spaceflight-induce immune system changes.

Bed rest, both with and without head-down tilt, has been extensively used as an earth-bound analog to study physiologic effects mimicking those occurring in weightlessness during spaceflight. We have been able to show in six subjects that 4 weeks of head-down tilt bed rest induces a significant decrease in interleukin-2 secretion by PHA-stimulated T lymphocytes. Another study, lasting 113 days, with two subjects showed a decreased interleukin-2 receptor expression in PHA-stimulated peripheral blood mononuclear cells but a decreased interleukin-2 production in one subject only. Under the same conditions, interleukin-1 production was largely increased in both subjects. Several other immune parameters were also analyzed. Increased interleukin-1 production could contribute to bone mineral loss encountered during bed rest and decreased interleukin-2 secretion could play a role in the appearance of infectious diseases often observed during bed red.

Antibody Formation↗
PubMed1996

Longitudinal study of FM system use in nonacademic settings: effects on language development.

OBJECTIVE: The overall objective of the present study was to assess the efficacy of FM system use in the home setting for a group of preschool children with mild-to-severe sensorineural hearing loss. Changes in language acquisition were monitored and compared with similar measures from a group of children who used hearing aids. Secondarily, the perceived benefits and practical problems associated with FM system use across a variety of nonacademic situations were documented. DESIGN: Ten children with mild-to-severe sensorineural hearing loss participated in a 2-yr longitudinal study investigating the efficacy of FM system use in the home setting. The subjects were divided into two groups: one group was instructed to use FM systems at home as often as possible while the other used only their personal hearing aids. Changes in language acquisition were monitored in both groups. Subjective benefit and the practical problems associated with use of FM systems outside of traditional academic environments were monitored via daily use logs, a weekly observation inventory, and a situational listening profile. RESULTS: The majority of children in both groups improved in all measures of language development over the study interval. Although there were relatively large individual differences in performance for some measures, no statistically significant differences between the FM and hearing aid users were found. However, some children in the FM group made unusually large gains in some aspects of language development over the study interval. In addition, both parents and children reported benefits of FM system use in specific listening situations. Throughout the 2-yr study, a number of practical problems associated with FM system use outside the classroom were identified. CONCLUSIONS: Formal language measures did not yield significant differences between the FM and HA groups, but some subjects had rates of language acquisition which suggested that FM system use may be beneficial in selected cases. In addition, subjective reports of FM system benefit suggest that appropriate use of the device may facilitate effective communication in a variety of listening situations. Although recent advances in FM system design may minimize some of the factors that reportedly restricted consistent FM use in this study, the complexities associated with the modes of operation and problems with FM interference remain issues that require consistent audiologic monitoring of FM system use in nonacademic environments.

Audiometry, Pure-Tone↗
PubMed1996

Measures of hearing aid gain for real speech.

OBJECTIVE: Non-speech stimuli typically are used to estimate the electroacoustic characteristics of a hearing aid. At present, there is no consensus as to what type of input stimulus will best represent the gain for real speech. The purpose of this study was to measure hearing aid gain using continuous discourse and to compare these values with gain measured with five different types of simpler stimuli. DESIGN: Hearing aid gain as a function of frequency was measured in a 2 cm3 coupler for 20 commercially available hearing aids. Circuitry included features such as linear peak clipping, compression limiting, 1-, 2-, and 3-channel full dynamic range compression, and adaptive compression. Input stimuli were a) 15 sec of continuous discourse, b) swept pure tones (SPTs), c) speech weighted composite noise (SWCN), d) simulated speech, e) speech weighted warble tones, and f) speech modulated noise. Input levels ranged from 50 to 80 dB SPL. RESULTS: In general, both SPTs and SWCN tended to underestimate the high-frequency gain for real speech. These discrepancies increased as a function of input intensity. On average, the SPT produced the greatest departure from the gain for real speech, producing differences for individual hearing aids as large as 10 to 14 dB. An analysis by circuit type revealed that discrepancies most likely occurred when a hearing aid was operating in a nonlinear mode. Of the five non-speech stimuli used, speech modulated noise and simulated speech seemed to provide the closest approximation to the gain measured with continuous discourse. CONCLUSIONS: When a hearing aid is operating in a nonlinear mode, non-speech stimuli will tend to underestimate the gain for real speech, particularly in the high frequencies. Under some conditions, these discrepancies may impact clinical decisions during the hearing aid fitting process. Additional studies are needed to elucidate the factors that contribute to the gain discrepancies observed in this study and to explore the use of additional stimuli (including short speech samples), which may result in better predictions of the gain for speech.

Correction of Hearing Impairment↗
PubMed1995

Two related potent antiviral compounds: 3-bromo-N-butyl-4-butylamino-1,8-naphthalenedicarboximide (1) and 4-amino-3-bromo-N-butyl-1,8-naphthalenedicarboximide (2).

The crystal structures of two brominated derivatives of 4-amino-N-butyl-1,8-naphthalenedicarboximide [alternative systematic names: 8-bromo-2-butyl-7-butylamino-1H-benzo[de]isoquinoline-1,3(2H)-dione, C20H23BrN2O2, and 7-amino-8-bromo-2-butyl-1H-benzo[de]isoquinoline-1,3(2H)-dione, C16H15BrN2O2] are reported. The N-butyl groups, which are common to both compounds, are perpendicular to the plane of the naphthalenedicarboximide ring system. The butylamino group at position 4 in (1) is in the plane of the naphthalenedicarboximide ring, with the butyl group oriented towards the peri position of the naphthalenedicarboximide ring. This arrangement results in an opening of the (ring C)-N-(butyl C) angle.

Antiviral Agents↗
PubMed1995

Isolating fetal nucleated red blood cells from maternal blood: the Baylor experience--1995.

In our previous work we have isolated fetal cells from maternal blood and used fluorescent in situ hybridization (FISH) for chromosome-specific probes to detect aneuploidy. Current efforts in the Baylor College of Medicine programme are focusing on obtaining consistency in flow-sorting methodology and on determining sensitivity and specificity. To this end, systematic evaluation of five glycophorin A (gly A) antibodies all produced agglutination, leading us to abandon the use of gly A antibodies for positive selection of fetal cells. Conversely, we have found LDS-751 to be useful for nuclear selection. CD45 negative selection can best be accomplished by the use of flasks coated with goat antibodies against mouse antibodies. Positive selection by flow sorting for either CD71+ cells or gamma-globin-positive cells seems to be successful. Using these two approaches, we have recently detected male (fetal) cells in pregnancies in which the fetus was 46,XY in 10 of 18 and in 12 of 14 cases, respectively.

Cell Separation↗
PubMed1995

Detection of low-grade mosaicism in fetal cells isolated from maternal blood.

Recovering and analysing fetal erythrocytes from maternal blood is being pursued for non-invasive prenatal genetic diagnosis. We report the observation of 46,XY/47,XXY mosaicism in fetal cells from a woman whose first-trimester chorionic villus sampling (CVS) initially showed only 46,XY. Only after exhaustive (500 cells) analysis were four XXY cells found in cultured villi.

Adult↗
PubMed1995

Excess intrauterine fetal demise associated with maternal human immunodeficiency virus infection.

A prospective study of transplacental transmission of human immunodeficiency virus (HIV) showed an increased rate of spontaneous fetal demise in HIV-seropositive mothers: 14 losses in 124 pregnancies. HIV was detected in placental and fetal tissues in 7 of 14 by in situ hybridization. The proportion of fetal infection far exceeded the transmission rate of 13% in liveborn babies. No association was seen between fetal transmission and a maternal history of drug abuse or coinfections; mothers with AIDS more often had fetal loss associated with HIV transmission than did asymptomatic mothers. In affected fetuses, HIV was detected in many tissues and was associated with thymic pathology. This suggests that maternal HIV infection increases the risk for pregnancy loss associated with HIV transmission. The possibility that HIV may be fetotoxic, that thymic dysfunction may interfere with pregnancy progression, or that the intrauterine milieu in HIV-seropositive pregnancies may be unfavorable (or a combination of factors) should be considered.

Adolescent↗
PubMed1995

The perception of amplified speech by listeners with hearing loss: acoustic correlates.

Audibility-based approaches to hearing-aid selection generally have focused on the long-term average speech spectrum (LTASS). Advances in amplification technology (e.g., multiband signal processing, level-dependent frequency shaping, full dynamic range compression, adaptive compression) make it difficult to predict the audibility of short-term components of speech from the amplified LTASS. This study was designed to quantify the audibility of a specific phonemes as processed by two different hearing-aid circuits (linear and full dynamic range compression), and to investigate the relation between audibility and performance on a nonsense syllable recognition task. Data were obtained from three subjects with moderate sensorineural hearing loss. Nine unvoiced consonants were presented in two vowel contexts (/i/ and /a/) in both the pre- and post-vocalic position at three intensities. While the performance on selected conditions appeared to vary by hearing-aid type, only one subject showed a statistically significant difference between the two hearing-aid systems. Acoustic analyses revealed a variety of spectral and temporal changes to the speech signal following processing. Estimates of audibility were based upon each subject's thresholds and an acoustic analysis of the amplified signal that varied across phonemes and consonant position. A signal detection approach was used to predict performance from a simple measure of audibility.

Amplifiers, Electronic↗
PubMed1995

Sensitivity of immune complex-dissociated p24 antigen testing for early detection of human immunodeficiency virus in infants.

Several investigators have suggested that early diagnosis of human immunodeficiency virus (HIV) infection in infants could be accomplished with a modified, more-sensitive, acid-dissociated p24 antigen enzyme-linked immunosorbent assay (ELISA) technique (p24 antigen immune complex dissociation [ICD]). We compared detection of HIV infection by HIV culture, PCR, and p24 antigen ICD assays in 46 infants by using samples collected independently. The detection sensitivity of the p24 antigen ICD assay was 0% with cord blood samples (2 HIV-positive infants), 38% with plasma samples from infants under 3 months of age (8 HIV-positive infants), and 58% overall (12 HIV-positive infants). By contrast, the sensitivities of HIV culture and PCR were 50% for cord blood samples, 75% for plasma samples from infants under 3 months of age, and 83% overall. These results indicate that the p24 antigen ICD does not offer the sensitivity necessary for this assay to be used as an indicator of HIV infection in infants.

Antigen-Antibody Complex↗
PubMed1995

Early detection of HIV in infants: aspects of design and analysis for diagnostic test studies.

OBJECTIVE: To review investigations on the early detection of HIV infection in infants to determine adherence to traditional methods of study design and analysis for evaluating new laboratory tests. DATA SOURCES: A National Library of Medicine (MEDLINE) search was conducted to identify such investigations through 1993. Cited references in identified manuscripts were also considered. The search was restricted to investigations of human subjects and those published in the English language. STUDY SELECTION: Final inclusion criteria included (1) report of the age and human immunodeficiency virus (HIV) infection status of the subjects at the time of the diagnostic testing, and (2) presentation of data allowing confirmation of presented analyses and additional analyses. DATA EXTRACTION: Criteria for judging the investigations included (1) whether the criteria used to determine the positive and negative test results were defined; (2) whether the necessary sample size for the study was calculated; (3) whether the patients studied were representative of the patients to whom the test would be applied; (4) whether a gold standard evaluation was performed; (5) whether the outcomes included in the analyses were independent; (6) whether the test characteristics were properly analyzed; and, (7) whether confidence intervals were presented. DATA SYNTHESIS: An informative presentation of a diagnostic test should include as a minimum the seven criteria listed above. Only 21 of 36 (58%) of the studies incorporated at least three of the criteria. CONCLUSIONS: There is a wide variation in the manner in which investigations of diagnostic tests are conducted and the results reported. Increased awareness and use of standard study designs and analyses will allow the application of metanalyses. Such analyses will help guide the direction taken for finding and establishing early diagnostic procedures for HIV infection at birth or during infancy.

AIDS Serodiagnosis↗
PubMed1995

Comparison of DNA fragmentation and color thresholding for objective quantitation of apoptotic cells.

Apoptosis is a process of cell death characterized by distinctive morphological changes and fragmentation of cellular DNA. Using video imaging and color thresholding techniques, we objectively quantitated the number of cultured CD4+ T-lymphoblastoid cells (HUT78 cells, RH9 subclone) displaying morphological signs of apoptosis before and after exposure to gamma-irradiation. The numbers of apoptotic cells measured by objective video imaging techniques were compared to numbers of apoptotic cells measured in the same samples by sensitive apoptotic assays that quantitate DNA fragmentation. DNA fragmentation assays gave consistently higher values compared with the video imaging assays that measured morphological changes associated with apoptosis. These results suggest that substantial DNA fragmentation can precede or occur in the absence of the morphological changes which are associated with apoptosis in gamma-irradiated RH9 cells.

Apoptosis↗