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Biomedical subjects

J E Butler

Publications and source records attributed to J E Butler.

At least 37 records · Page 2Linked to original sources

Diamond optically transparent electrodes: demonstration of concept with ferri/ferrocyanide and methyl viologen.

A new type of optically transparent electrode is reported on--an electrically conductive diamond thin film. The electrode was free-standing (0.38 mm thick and 8 mm in diameter), mechanically polished to a 7-nm rms roughness over a 10-microm linear distance, boron-doped (0.05% B/C in the reactant gas mixture), and mounted in a thinlayer transmission cell. The electrode has a short-wavelength cutoff of approximately 225 nm, which is the indirect band gap of the material, and transmits light out to at least 1,000 nm. In theory, the electrode has an optical window from 225 nm well out into the far-infrared, except for the boron acceptor band and the intrinsic multiphonon absorptions. The electrode was used to electrooxidize ferrocyanide to ferricyanide, and the absorbance change associated with the formation of the oxidized product (lambdamax = 420 nm) was spectroscopically monitored. The electrode was also used to electroreduce methyl viologen (MV2+) to the cation radical (MV+*) and the neutral (MV0). The depletion of MV2+ (lambdamax = 257 nm) and formation of MV+* (lambdama = 398 and 605 nm) were spectroscopically monitored.

Journal Article↗

Gender-related differences of hyaluronic acid distribution in the human vocal fold.

OBJECTIVE: To quantify the distribution of hyaluronic acid (HA) within the normal human vocal fold and evaluate gender- and age-related differences. STUDY DESIGN: Prospective, cross-sectional study by age and gender groups using cadaveric vocal folds to study hyaluronic acid differences. METHODS: Twenty-five vocal fold halves obtained at autopsy were evaluated from nine adult male (age range, 34-52 y), seven adult female (age range, 21-41 y), four geriatric male (age range, 65-77 y), and five geriatric female (age range 65-82 y) subjects. Midmembranous vocal fold sections were stained for HA using the Muller-Mowry colloidal iron (acid mucopolysaccharide) stain with and without hyaluronidase. Digital imaging software was used to quantify the distribution of HA across the lamina propria at the vocal fold edge from superficial (subepithelial) to deep (vocalis muscle). RESULTS: A significant gender difference (P =.03) was noted in distribution when controlled for age. Combined, male subjects had a relatively constant distribution pattern throughout the depth of lamina propria. Female subjects showed relatively less HA in the first 15% of depth (most superficial) but showed more HA in the deeper 40% to 100% (toward vocalis muscle) than in male subjects. CONCLUSIONS: Important clinical implications may be extrapolated from our findings. First, relatively less HA in the most superficial area implies less protection from vibratory trauma and overuse and may explain in part why more female than male patients have phonotrauma to phonotraumatic lesions. Second, evaluating the distribution of HA in the normal human vocal fold allows more direction in our application of HA as a therapeutic lamina propria substitute in patients with voice disorders.

Adult↗

Discharge frequencies of single motor units in human diaphragm and parasternal muscles in lying and standing.

Single motor unit discharge was measured directly in diaphragm and parasternal intercostal muscles to determine whether neural drive to human inspiratory muscles changes between lying and standing. The final discharge frequency of diaphragmatic motor units increased slightly, by 1 Hz (12%; P < 0.01), when subjects were standing [182 units, median 9.1 Hz (interquartile range 7.6-11.3 Hz)] compared with lying supine [159 units, 8.1 Hz (6.6-10.3 Hz)]. However, this increase with standing occurred in only two of six subjects, in one of whom tidal volume increased significantly during standing. Parasternal intercostal motor unit final discharge frequencies did not differ between standing [116 units, 8.0 Hz (6.6-9.6 Hz)] and lying [124 units, 8.4 Hz (7.0-10.3 Hz)]. The discharge frequencies at the onset of inspiration did not differ between lying and standing for either muscle. A larger proportion of motor units in both inspiratory muscles had postinspiratory or tonic expiratory activity for lying compared with standing (15 vs. 4%; P < 0.05). We conclude that there is no major difference in the phasic inspiratory drive to the diaphragm with the change in posture.

Diaphragm↗

Sacroiliac joint dysfunction: a long-term follow-up study.

Sixty-nine patients with sacroiliac joint dysfunction were prospectively evaluated and treated with a structured physical therapy program. Follow-up clinical outcome was obtained from a patient questionnaire administered by an independent reviewer a minimum of 2 years after treatment. Average patient age was 40 years, and 80% were women. Ninety-five percent rated their result as good or excellent, while 5% believed their outcome was fair or poor. A structured physical therapy program can produce good long-term results in most patients; however, 5% continue to be symptomatic. This small subset may be candidates for more invasive evaluation.

Adolescent↗

Antibody repertoire development in fetal and neonatal piglets. II. Characterization of heavy chain complementarity-determining region 3 diversity in the developing fetus.

Since the actual combinatorial diversity in the V(H) repertoire in fetal piglets represents <1% of the potential in mice and humans, we wondered whether 1) complementarity-determining region 3 (CDR3) diversity was also restricted; 2) CDR3 diversity changed with fetal age; and 3) to what extent CDR3 contributed to the preimmune VDJ repertoire. CDR3 spectratyping and sequence analyses of 213 CDR3s recovered from >30 fetal animals of different ages showed that >95% of VDJ diversity resulted from junctional diversity. Unlike sheep and cattle, somatic hypermutation does not contribute to the repertoire. These studies also revealed that 1) N region additions are as extensive in VDJ rearrangements recovered at 30 days as those in late term fetuses, suggesting that TdT is fully active at the onset of VDJ rearrangement; 2) nearly 90% of all rearrangement are in-frame until late gestation; 3) the oligoclonal CDR3 spectratype of 30-day fetal liver becomes polyclonal by 50 days, while this change occurs much later in spleen; 4) there is little evidence of individual variation in CDR3 spectratype or differences in spectratype among lymphoid tissues with the exception of the thymus; and 4) there is a tendency for usage of the most J(H) proximal D(H) segment (D(H)B) to decrease in older fetuses and for the longer D(H) segment to be trimmed to the same length as the shorter D(H) when used in CDR3. These findings suggest that in the fetal piglet, highly restricted combinatorial diversity and the lack of somatic mutation are compensated by early onset of TdT activity and other mechanisms that contribute to CDR3 junctional diversity.

Animals↗

Early ontogeny of thymocytes in pigs: sequential colonization of the thymus by T cell progenitors.

Successive colonization of the thymus by waves of thymocyte progenitors has been described in chicken-quail chimeras and suggested from studies in mice. In swine, we show that the first CD3epsilon-bearing thymocytes appear on day 40 of gestation (DG40). These early thymocytes were CD3epsilonhigh and belonged to the gammadelta T cell lineage. Mature CD3epsilonhigh alphabeta thymocytes were observed 15 days later (DG55), and their occurrence was preceded by the appearance of CD3epsilonlow thymocytes (DG45). Thereafter, we observed transient changes in thymocyte subset composition (DG56-DG74), which can be explained by a gap in pro-T cell delivery to the thymus. This delivery gap corresponds with the expression of the pan-leukocyte CD45 and pan-myelomonocytic SWC3a markers in fetal liver and bone marrow and is probably caused by shifting of primary lymphopoiesis between these organs. Therefore, we conclude that the embryonic thymus is colonized by at least two successive waves of hemopoietic progenitors during embryogenesis and that the influx of thymocyte progenitors is discontinuous. Surface immunophenotyping and cell cycle analysis of thymocyte subsets allowed us to compare thymocyte differentiation in pigs with that described for rodents and humans and to propose a model for T cell lymphopoiesis in swine. We also observed that the porcine IL-2Ralpha (CD25), a typical differentiation marker of pre-T cells in mice and humans, was not expressed on thymocyte precursors in pigs and could only be found on mature thymocytes. Finally, we observed a subset of TCRgammadelta+ thymocytes that were cycling late during their development in the thymus.

Animals↗

Ischaemia after exercise does not reduce responses of human motoneurones to cortical or corticospinal tract stimulation.

Motor unit firing rates and voluntary activation of muscle decline during sustained isometric contractions. After exercise, the responses to motor cortical and corticospinal stimulation are reduced. These changes may reflect motoneuronal inhibition mediated by group III and IV muscle afferents. To determine whether the post-contraction depression of the responses to corticospinal or motor cortical stimulation could be maintained by continued firing of ischaemically sensitive group III and IV muscle afferents, we examined responses in muscles that were held ischaemic after exercise. Following a sustained maximal voluntary contraction (MVC) of the elbow flexors lasting 2 min, the response to stimulation of the corticospinal tract was reduced but the usual recovery (over approximately 2 min) was not delayed when the muscles were maintained ischaemic for 2 min after the contraction. Following a sustained MVC, the time course of the reduction in the response to motor cortical stimulation (a gradual decrease over approximately 2 min, maintained for > 10 min) was also not altered if the muscle was held ischaemic. Mean arterial blood pressure rose to 155 +/- 12 mmHg during the 2 min MVC, declined to 125 +/- 9 mmHg immediately after it, but remained at this level without returning to pre-exercise levels (102 +/- 10 mmHg) until circulation to the arm was restored. This confirms that the sustained MVC activated a reflex dependent on group III and IV muscle afferents. This study shows that ischaemically sensitive group III and IV muscle afferents do not mediate depression of responses to motor cortical or corticospinal stimulation after fatiguing exercise. It also suggests that firing of such afferents does not directly inhibit motoneurones or motor cortical output cells.

Adult↗

Genetic differences in the frequency of the hinge variants of porcine IgA is breed dependent.

The distribution of the IgA(a) and IgA(b) alleles of porcine IgA in over 160 randomly-selected animals revealed an abundance of heterozygotes but only two b/b homozygotes. Since the IgA(b) allotype is a splice site mutant lacking two-thirds of the hinge, this study tests the hypothesis that pigs with this genotype may be at a selective disadvantage while heterozygous individuals may be at some advantage. This hypothesis was tested by collecting data on 374 animals of known breed and often parentage. We show here that when breed was not considered, young animals of known parentage had genotypic frequencies identical to that expected for Mendelian alleles but that a/b heterozygotes were overrepresented in adults. However, when analyzed with regard to breed, a very strong association between breed and the frequency of the IgA(a) and IgA(b) alleles was discovered. Meishan and NIH minipigs were homozygous for IgA while heterozygotes predominated in Berkshire, Chester White, Durocs, Hampshire and Landrace. Animals homozygous for IgA(b) were best represented in the White Cross line. We show here that this very strong breed dependency of IgA allotypy in swine can produce a sample bias that can explain why only two b/b homozygotes (1.3%) were found in the 160 randomly-selected samples since the original samples came from primarily Landrace and Yorkshire animals. The expected frequency of b/b homozygotes in these breeds would be <3%. Thus, the data presented here reject the hypothesis that swine homozygous for a trait that results in loss of two-thirds of the IgA hinge, are selected against and that heterozygotes are positively selected. Rather, the study shows that IgA(a) and IgA(b) appear to be simple, breed-dependent allotypic markers.

Alleles↗

Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays.

A very large proportion of modern immunoassays involve the use of synthetic solid phases to immobilize one of the reactants. These solid-phase immunoassays (SPIs) therefore involve ligand-receptor interactions that occur within a reaction volume close to the solution/solid phase interface. As a consequence, the immunochemistry/biochemistry of these ligand-receptor interactions differs from that of their counterparts in solution. Furthermore, the immobilization process can significantly alter the biological activity of the reactant; most adsorbed proteins on polystyrene or silicone are partially or largely denatured. Therefore the use of alternative methods of immobilization is attractive but may result in little increase in the amount of total functional reactant. However, all commonly used solid phases do not have the same properties or the same capacity for reactant immobilization or experience the same level of nonspecific binding. Empiricism plays a major role in SPIs. Derivations of mass law equations for measuring the antigen capture of solid-phase antibodies, for determining the affinity of solid phase for protein adsorption, and for estimating antibody affinity are reviewed.

Adsorption↗

Changes in muscle afferents, motoneurons and motor drive during muscle fatigue.

Fatigue is a reduction of maximal muscle force or power that occurs with exercise. It is accompanied by changes at multiple levels in the motor pathway and also by changes in the discharge patterns of muscle afferents. Changes in afferent firing can lead to altered perceptions and can also act on the efferent pathway. Changes in the motor pathway include slowing of motor unit firing rates during sustained maximal voluntary contractions (MVCs). Muscle responses to stimulation at different levels of the motor pathway also change. Transcranial magnetic stimulation of the motor cortex and stimulation of descending tracts in the spinal cord in human subjects show an increase in the response of the cortex and a decrease in response of the motoneuron pool during sustained MVCs. In addition, the silent period following magnetic stimulation is prolonged. During relaxation after fatiguing exercise, muscle responses to stimulation of the motor cortex are initially facilitated and are then depressed for many minutes, whereas responses to descending tract stimulation are initially depressed but recover over about 2 min. Although some of the loss of force of fatigue does occur through inadequate drive to the muscle, it is not clear which, if any, of the changes described in the cortex or the motoneurons are responsible for loss of maximal voluntary force and thus contribute to fatigue. Changes may be associated with muscle fatigue without causing it.

Animals↗

Porcine IgA allotypes are not equally transcribed or expressed in heterozygous swine.

The prediction of 1:1 expression of constant region allotypes in heterozygous animals assumes that productive VDJ rearrangements occur at random between chromosomes, switch recombination is random, there is no allele-related defect in switching and there is no selection for a B-cell receptor bearing a certain constant region allotype. In data reported here, this prediction was often not fulfilled for the transcripts encoding the IgAa and IgAb alleles of porcine IgA including those from late term fetal piglets that are not in contact with environmental antigens or maternal regulatory factors. In the spleen, thymus, mesenteric lymph node (MLN), ileal Peyer patches, parotid gland and PBLs of 5-week-old conventionally-reared Duroc pigs, ratios of IgAa to IgAb transcripts as high as 4:1 were observed. Since White Cross animals had significantly higher levels of IgAb than IgAa (some >3-fold), a allele-linked switch defect cannot explain the deviation from the expected 1:1 ratio. When the IgAa:IgAb ratios in older Durocs and those reared at a different site were studied, no evidence for breed dependence of differential transcription was found. Total serum IgA levels paralleled total transcript levels in PBLs while particularly in White Cross animals, the IgAa:IgAb ratio in serum was higher in many animals than the IgAa:IgAb transcript ratio in their PBLs. We conclude that deviations from the expected 1:1 ratio of allotype transcripts and secreted IgA in young pigs is normal and deviations from this ratio also occur during fetal life in the absence of environmental antigens and maternal regulatory factors. We speculate that postnatal deviations result from: (A) exposure to environmental antigens that selectively expand B-cells expressing V(H) gene alleles linked to either IgAa or IgAb or (B) some form of colostrum-dependent regulation. Pre-natal regulation may depend on the selection of B-cells bearing certain V(H) or C(H) encoded BCRs by stromal ligands such as fetal B-cell superantigens.

Animals↗

Antibody repertoire development in fetal and neonatal piglets. V. VDJ gene chimeras resembling gene conversion products are generated at high frequency by PCR in vitro.

The recovery of VDJ rearrangement is most often accomplished by PCR amplification of DNA extracted from mixtures of B-cells. Using this procedure in swine, VDJs containing chimeric V(H) genes that resemble gene-conversion products, are frequently encountered. To examine whether these chimeras could be the result of PCR artifacts, we used different combinations of swine VDJ templates, each having unique CDR1, CDR2 and D(H) segments, to generate >2600 clones. Using equal amounts of two templates and 30 cycles of PCR, up to 45% of the resultant clones were VDJ chimeras. The frequency of chimeras was independent of the specific VDJ template and the chimeras were generated regardless of whether Taq-, Pfu- or mixtures of Taq- and Pfu-polymerases were employed or whether PCR extension time was prolonged six-fold. The frequency of generating chimeras was dependent on the ratio of the two target DNAs although even ratios approximately 1:10 generated approximately 10% chimeric VDJs. Chimeras could be generated using only 10 cycles of PCR or using the initial template DNAs diluted as much as 1:10000. Of the 279 chimeric VDJs generated, 61% of the crossovers occurred in FR3, 21% in FR2 and 18% in both FR2 and FR3. We interpret these results to mean that in vivo gene conversion in this species can only be unambiguously proven when the VDJs from individual B-cells are bearing a single VDJ rearrangement amplified and sequenced or when VDJs are cloned without the use of PCR.

Animals↗

Extended and Point Defects in Diamond Studied with the Aid of Various Forms of Microscopy.

It is shown that star disclinations can be a significant source of stress in chemical vapor deposited (CVD) diamond. This purely geometrical origin contrasts with other sources of stress that have been proposed previously. The effectiveness is demonstrated of the use of electron irradiation using a transmission electron microscope (TEM) to displace atoms from their equilibrium sites to investigate intrinsic defects and impurities in CVD diamond. After irradiation, the samples are studied by low temperature photoluminescence microscopy using UV or blue laser illumination. Results are given that are interpreted as arising from isolated <100> split self-interstitials and positively charged single vacancies. Negatively charged single vacancies can also be revealed by this technique. Nitrogen and boron impurities may also be studied similarly. In addition, a newly developed liquid gallium source scanned ion beam mass spectrometry (SIMS) instrument has been used to map out the B distribution in B doped CVD diamond specimens. The results are supported by micro-Raman spectroscopy.

Journal Article↗

Antibody repertoire development in fetal and newborn piglets, III. Colonization of the gastrointestinal tract selectively diversifies the preimmune repertoire in mucosal lymphoid tissues.

Changes in the VH-region repertoire of isolator piglets reared for 6 weeks under germ-free (GF) conditions and those colonized (COL) with a defined exclusion flora on the 1st day of life were compared. Although serum immunoglobulin levels were 20-100-fold higher in COL piglets than GF piglets, an analysis of peripheral blood B cells (PBBs) indicated that: GF and COL piglets used the same four VH genes and two DH segments during the 6-week period; proportional usage of VH genes and DH segments was the same as in fetal animals; and VH and DH usage did not differ between COL and GF animals. This pattern differed from the PBBs from 6-week-old conventional (CONV) piglets. When the sequences of 73 splenic CDR3 segments were analysed, DH usage and mutation frequency were the same in sequences from both 6-week-old GF and COL piglets; mutations were infrequent and occurred with the same frequency as in 110-day fetal spleen. However, the median CDR3 length in COL piglets was shifted upward due to 3' DH N-nucleotide additions. Neither COL nor GF animals made specific serum antibodies to phosphoryl choline given parenterally on a T-cell dependent carrier. In contrast to the near absence of a colonization effect in PBBs and splenic DNA, rearranged variable heavy-chain gene segments (VDJs) recovered from the DNA of mucosal lymphoid tissues of COL piglets showed pronounced differences from those recovered from GF animals in usage of DHA-, DHB-and VHB- and in the frequency of point mutation. The mucosal VDJ transcripts and those from the spleen were similarly affected by colonization. This effect on mucosal lymphoid tissue was consistent with the five-fold selective increase in serum immunoglobulin A (IgA) levels relative to IgM and IgG. Comparison of IgM and IgA transcripts from mucosal tissues suggested that IgA and IgM clones diversify in parallel. Our findings are the first to show that colonization of the gastrointestinal tract of offspring separated from their mothers, differs from 'conventionalized' GF animals in that colonization preferentially influences diversification and expansion of the preimmune IgM and IgA repertoire in mucosal lymphoid tissues but not in PBBs and seldom/modestly in VDJs from splenic DNA.

ATP-Binding Cassette Transporters↗

Supraspinal fatigue during intermittent maximal voluntary contractions of the human elbow flexors.

Responses to transcranial magnetic stimulation in human subjects (n = 9) were studied during series of intermittent isometric maximal voluntary contractions (MVCs) of the elbow. Stimuli were given during MVCs in four fatigue protocols with different duty cycles. As maximal voluntary torque fell during each protocol, the torque increment evoked by cortical stimulation increased from approximately 1.5 to 7% of ongoing torque. Thus "supraspinal" fatigue developed in each protocol. The motor evoked potential (MEP) and silent period in the elbow flexor muscles also changed. The silent period lengthened by 20-75 ms (lowest to highest duty cycle protocol) and recovered significantly with a 5-s rest. The MEP increased in area by >50% in all protocols and recovered significantly with 10 s, but not 5 s, of rest. These changes are similar to those during sustained MVC. The central fatigue demonstrated by the torque increments evoked by the stimuli did not parallel the changes in the electromyogram responses. This suggests that part of the fatigue developed during intermittent exercise is "upstream" of the motor cortex.

Adult↗

Age- and gender-related collagen distribution in human vocal folds.

The composition of the lamina propria in human vocal folds has been shown to affect vocal performance. Collagen plays a significant role in the biomechanical effects of the lamina propria. Specifically, it lends tensile strength to the rapidly oscillating fold. We obtained from a state medical examiner 38 larynges from men and women in infant, adult, and geriatric age groups. We stained the vocal folds for collagen using a picric acid stain and studied them using an image analysis system. Distributions of collagen were measured from the superficial to deep layers (from epithelium to vocalis muscle) within the lamina propria. The data showed an increase in collagen content from infant to adult stages. Infant folds had about 51% of the collagen found in all adults and in geriatric patients (p < .001). There was no significant difference between adult and geriatric folds (p < .16). There was, however, a gender difference in the amount of collagen in both adult and geriatric specimens. Female adult and geriatric folds had about 59% of the collagen found in male adult and geriatric folds (p < .001). The distribution pattern of collagen showed that most of the collagen was present in the deep layer. From these data we conclude that there are age-related and gender-related differences between male and female infant, adult, and geriatric vocal folds. Stress-strain performance studies need to be correlated with histologic findings to better study the phonetic implications of these findings.

Adult↗

Tritrichomonas foetus extracellular cysteine proteinase cleavage of bovine IgG2 allotypes.

Bovine trichomoniasis is a sexually transmitted disease associated with reproductive failure. Systemic immunization results in protective IgG antibodies in uterine and vaginal secretions. Because bovine IgG2 is a better opsonin than IgG1, it is potentially important in defense. Yet, Tritrichomonas foetus extracellular cysteine proteinase (TFECP) cleaves bovine IgG2, evading protective IgG2 responses. Variations in resistance of the 2 IgG2 allotypes to digestion may explain inherited differences in protection. To address this hypothesis, TFECP was incubated with both IgG2 allotypes at different concentrations and times. The digestion products were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, stained, and quantitated by image analysis. IgG2a was digested faster by TFECP than IgG2b. Differences in the sizes and numbers of digestion products were observed, but the presence of bands the size of Fc and Fd fragments indicated that both allotypes were cleaved at the hinge. Cysteine in the digestion mixture reduced the antibody molecules and increased the rate of digestion, but IgG2a was still more susceptible to cleavage than IgG2b in the absence of cysteine. Thus, not only reduced H chains can be cleaved by cysteine proteinase secreted by T. foetus but also intact functional antibody molecules. Because parasites may evade protective antibody responses by cleaving IgG2, animals with the more resistant IgG2b allotype may be better protected by immunization than animals with the more readily digested IgG2a allotype.

Animals↗

Impaired response of human motoneurones to corticospinal stimulation after voluntary exercise.

1. Activation of descending corticospinal tracts with transmastoid electrical stimuli has been used to assess changes in the behaviour of motoneurones after voluntary contractions. Stimuli were delivered before and after maximal voluntary isometric contractions (MVCs) of the elbow flexor muscles. 2. Following a sustained MVC of the elbow flexors lasting 5-120 s there was an immediate reduction of the response to transmastoid stimulation to about half of the control value. The response recovered to control levels after about 2 min. This was evident even when the size of the responses was adjusted to accommodate changes in the maximal muscle action potential (assessed with supramaximal stimuli at the brachial plexus). 3. To determine whether the post-contraction depression required activity in descending motor paths, motoneurones were activated by supramaximal tetanic stimulation of the musculocutaneous nerve for 10 s. This did not depress the response to transmastoid stimulation. 4. Following a sustained MVC of 120 s duration, the response to transcranial magnetic stimulation of the motor cortex gradually declined to a minimal level by about 2 min and remained depressed for more than 10 min. 5. Additional studies were performed to check that the activation of descending tracts by transmastoid stimulation was likely to involve excitation of direct corticospinal paths. When magnetic cortical stimuli and transmastoid stimuli were timed appropriately, the response to magnetic cortical stimulation could be largely occluded. 6. This study describes a novel depression of effectiveness of corticospinal actions on human motoneurones. This depression may involve the corticomotoneuronal synapse.

Adult↗