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Biomedical subjects

C A Smith

Publications and source records attributed to C A Smith.

At least 163 records · Page 9Linked to original sources

Sites of estrogen receptor and aromatase expression in the chicken embryo.

Estrogens have been implicated in sexual differentiation of both the gonads and the genitalia of birds. In chicken embryos, the gonads are steroidogenically active from an early age, and the aromatase gene, (cAROM), necessary for estrogen synthesis, is expressed only in females at the time of gonadal sex differentiation. However, no studies have directly demonstrated the distribution of estrogen receptor (cER) transcripts or proteins in the embryonic avian reproductive system. Whole-mount in situ hybridization and immunohistochemistry were used here to identify sites of estrogen receptor expression in the embryonic chicken urogenital system. Estrogen receptor mRNA was observed in both male and female gonads prior to morphological differentiation, at Stage 26 (4.5 days of incubation), and continued until after sexual dimorphism at Stage 32 (7.5 days). Transcripts of cER were also detected in the Müllerian ducts and developing external genitalia of both sexes. Estrogen receptor protein was analysed in the embryonic gonads by immunohistochemistry and found to be most abundant in the cortex of the left ovary, although it was also present in the medulla of both female gonads. No significant cER protein expression was detected in the male gonad by immunohistochemistry. In contrast, the aromatase gene was expressed in the gonads of female embryos from the onset of sexual dimorphism but was not detectable in male gonads at any stage examined. These findings suggest that estrogen involvement in both gonadogenesis and genital development in chickens is mediated by the estrogen receptor.

Animals↗

Beyond good and bad coping: a multidimensional examination of coping with pain in persons with rheumatoid arthritis.

This article examines the utility of using a multidimensional instrument to assess pain coping in two samples of persons with rheumatoid arthritis (total N = 378). The predictive validity of the newly developed Vanderbilt Multidimensional Pain Coping Inventory (VMPCI), which assesses eleven distinct coping strategies, was compared to that of the previously validated Vanderbilt Pain Management Inventory (VPMI), which assesses the two broad dimensions of active and passive coping. In both samples, the VMPCI demonstrated reliable incremental validity over the VPMI in predicting both positive and negative psychological adjustment, and the predictive ability of the VPMI was highly redundant with that of the VMPCI. Moreover, using the VMPCI, both positive and negative adjustment were related to multiple coping strategies in ways that could not be revealed by the VPMI. These results offer considerable validation to the VMPCI and illustrate the value of assessing coping multidimensionally.

Adaptation, Psychological↗

Pulmonary-locomotory interactions in exercising dogs and horses.

In exercising quadrupeds, limb movement is often coupled with breathing frequency. This finding has lead some investigators to conclude that locomotory forces, associated with foot plant, abdominal visceral displacements or lumbo-sacral flexion, are the primary determinants of airflow generation. Analysis of respiratory muscle electrical activation (EMG) and contraction profiles in chronically instrumented dogs and horses, along with measurements of esophageal pressure (Pes) changes and limb movements, provide evidence that each breath during the exercise hyperpnea is determined by respiratory neuromuscular events. Specifically: (1) Phasic diaphragmatic EMG and tidal shortening are always synchronous with decreases in Pes; (2) decrements in Pes are always associated with inspiratory flow generation; and (3) strict phase coupling between breathing and stride frequency is not obligatory. Thus, although locomotory-associated forces may minimally assist with flow generation, they are not the primary determinants of breathing during exercise.

Animals↗

Smooth muscle development in the obstructed fetal bladder.

OBJECTIVES: To evaluate changes in the smooth muscle and connective tissue development in the obstructed and normal fetal bladder. METHODS: The smooth muscle and connective tissue composition of 19 fetal urinary bladders, including those of 9 fetuses with anatomic obstruction and 10 controls free of urologic disease, were analyzed by light microscopy and computer-assisted color image analysis. RESULTS: The bladder wall thickness was markedly increased in obstructed fetuses throughout gestation as compared with that in controls. The disparity in bladder wall thickness increased rapidly during gestation. The percent area density of smooth muscle and connective tissue as well as the ratio of smooth muscle to connective tissue remained the same in the obstructed and normal control fetal bladders. CONCLUSIONS: Although bladder outlet obstruction is associated with a marked increase in bladder wall thickness, the percent of smooth muscle and connective tissue comprising the mural histology remains relatively constant as compared with that of normal fetal controls. This study suggests that bladder outlet obstruction in the fetus is not associated with excess collagen deposition but rather with an increased amount of bladder with normal cellular content and a concomitant increase in smooth muscle development.

Connective Tissue↗

Excision-repair patch lengths are similar for transcription-coupled repair and global genome repair in UV-irradiated human cells.

We have used the buoyant density shift method to measure excision-repair patch lengths in UV-irradiated repair-proficient human cells and in primary fibroblasts belonging to xeroderma pigmentosum complementation group C (XP-C), in which excision repair of UV-induced photoproducts is dependent upon transcription. The patch size was found to be about 30 nucleotides for both cell types. This agrees with the size of the DNA fragments excised in vitro by the dual incisions of the structure-specific nucleases XPG and ERCC1-XPF. We conclude that the XPC protein is not required to target the excision nucleases to sites of DNA cleavage in transcribed strands of expressed genes or to protect the newly incised DNA from further processing by exonucleases.

Cell Line↗

Gonadal sex differentiation in chicken embryos: expression of estrogen receptor and aromatase genes.

Estrogen is implicated in sexual differentiation of the avian gonad. Expression of the estrogen receptor and aromatase genes was therefore examined at the time of gonadal sex differentiation in chicken embryos, using reverse transcription and the polymerase chain reaction (RT-PCR). Estrogen receptor (cER) transcripts were detected in the gonads of both presumptive sexes at embryonic days 4.5, 5.5 and 6.5, and in female but not male urogenital tissues at day 3.5. Aromatase (cAROM) transcripts were detected in female but not male gonads from day 6.5 of embryogenesis, and in adult gonads of both sexes. Both female and male embryos thus express cER mRNA before morphological differentiation of the gonads, which begins on day 5, whereas cAROM expression begins at or shortly after the onset of differentiation and is female-specific. Examination of other tissues showed that, in 5.5-day-old embryos, cER expression was limited to the gonads; no transcripts were detected in the mesonephric kidney, liver, brain, hindlimb or heart of either sex. In 9.5-day-old female embryos, cER and cAROM transcripts were present in both the left (ovarian) and the right (regressing) gonads. Altogether, these observations imply that the gonads of both sexes develop the capacity to respond to estrogens early in embryogenesis, before morphological differentiation, whereas the capacity to synthesize estrogens is female-specific and occurs later, at the time of differentiation. These observations are consistent with estrogens having a key role in ovarian development.

Animals↗

The novel receptor TRAIL-R4 induces NF-kappaB and protects against TRAIL-mediated apoptosis, yet retains an incomplete death domain.

A fourth member of the emerging TRAIL receptor family, TRAIL-R4, has been cloned and characterized. TRAIL-R4 encodes a 386-amino acid protein with an extracellular domain showing 58%-70% identity to those of TRAIL-R1, TRAIL-R2, and TRAIL-R3. The signaling capacity of TRAIL-R4 is similar to that of TRAIL-R1 and TRAIL-R2 with respect to NF-kappaB activation, but differs in its inability to induce apoptosis. Yet TRAIL-R4 retains a C-terminal element containing one third of a consensus death domain motif. Transient overexpression of TRAIL-R4 in cells normally sensitive to TRAIL-mediated killing confers complete protection, suggesting that one function of TRAIL-R4 may be inhibition of TRAIL cytotoxicity. Like TRAIL-R1 and TRAIL-R2, this receptor shows widespread tissue expression. The human TRAIL-R4 gene has been mapped to chromosome 8p22-21, clustered with three other TRAIL receptors.

Amino Acid Sequence↗

Control of virus-induced lymphoproliferation: Epstein-Barr virus-induced lymphoproliferation and host immunity.

Epstein-Barr virus (EBV) is a latent herpesvirus that is associated with a number of tumors. EBV-infected cells show three patterns of latency ranging from type 1, where only one EBV-encoded antigen is expressed, to type 3, where all nine latent cycle proteins encoded by EBV are expressed. Malignancies exhibiting the type 3 latency pattern are highly immunogenic and occur only in immunocompromised patients. It has recently been shown that adoptive immunotherapy with EBV-specific cytotoxic T lymphocytes is an effective therapy for such tumors. Immunotherapy strategies and approaches to increase tumor immunogenicity are now being evaluated in tumors expressing type 2 latency.

Animals↗

Results of the Mitrofanoff procedure in urinary tract reconstruction in children.

OBJECTIVES: To evaluate the success and long-term complications associated with the use of continent catheterizable conduits based on the Mitrofanoff principle in children. PATIENTS AND METHODS: The records of 43 patients (21 female and 22 male) who underwent the construction of a continent catheterizable stoma based on the Mitrofanoff principle between 1987 and 1996 were reviewed retrospectively. The mean age at surgery was 10 years (range 3-21) and the mean follow-up was 3 years (range 0.5-6.5). Twenty-eight of the 43 children underwent augmentation cystoplasty in conjunction with the Mitrofanoff procedure, using ileum in 17, sigmoid in seven, caecum in two and stomach in one; detrusormyectomy was performed in one child. Fifteen patients had only a continent catheterizable stoma formed. The most common type of conduit was appendicovesicostomy (36 of 43 children): other conduits were constructed with ureter (four), tapered ileum (two) and fallopian tube (one). RESULTS: Stomal continence was achieved in 42 of 43 patients (98%). The most common late complication was difficulty in catheterization, which occurred in 14 patients (32%). Stomal prolapse requiring revision occurred in one patient (2%). Conduit dilatation was initially attempted in all patients with difficult catheterization, although it failed in 11 who then required surgical revision. Therefore, the overall revision rate was 28% (12 of 43). The site of stomal placement (umbilical or abdominal) did not significantly influence the risk of difficulty with catheterization. CONCLUSION: The Mitrofanoff procedure can simplify catheterization in children who are dependent upon intermittent catheterization. The vermiform appendix appears to be the best source for constructing the conduit. While stomal continence is excellent, conduit stenosis remains a frequent complication regardless of stomal location.

Adolescent↗

Factors associated with early sexual activity among urban adolescents.

This study uses lifespan and ecological frameworks to investigate the factors associated with early adolescent sexual activity. Data from a longitudinal study of urban teenagers of color address three issues: (1) the prevalence and pattern of sexual activity among boys and girls ages 15 and younger, (2) the link between early sexual activity and high-risk sexual behavior, and (3) the life contexts linked with early sexual activity. Results from 803 African American and Hispanic adolescents suggest a high prevalence of early sexual activity, which is associated with higher rates of childbearing and risky sexual behavior than sexual activity initiated in later adolescence. Somewhat different factors are associated with early sexual activity for boys and girls, although family composition, parent attachment, and substance use are important for both genders. Implications for intervention are discussed.

Adolescent↗

Interdisciplinary student health teams: combining medical education and service in a rural community-based experience.

Several initiatives have been introduced over the years to address the maldistribution of health care professionals and to improve access to care for underserved rural populations. One of these is the sponsorship of community-based, service-oriented teams comprised of students from various health disciplines. This study investigated extramural training as a complement to traditional hospital-based experiences. The specific objective of the study was to determine the extent to which the nation's medical schools combine training with a rural community-based experience in the form of an interdisciplinary student health team program. In the fall of 1994, a 32-item questionnaire was mailed to the chief academic or clinical affairs administrators of the nation's 126 allopathic medical schools. A total of 104 (82.5%) medical schools responded to the survey. Eighty-six of the respondents (82.7%) reported some type of rural training or public service activity; 22 (21.2%) acknowledged the sponsorship of an interdisciplinary student health team program. Small rural communities, those with populations of 5,000 or fewer, were the focus of 76 percent of the reporting programs. Nearly two-thirds of the reporting programs were located in the South, the region with the nation's lowest physician-to-population ratio. The nursing and medical professions were most frequently represented, although a wide range of disciplines were identified as participating on the student health teams. Activities of the teams included both ambulatory care and community outreach services. The majority of the programs used team-building exercises to enhance team effectiveness. Extramural training programs offer students a realistic examination of the social, cultural, economic, and political forces that influence both individual and community health. Rural community-based programs, such as interdisciplinary student health teams, should be valued because they can strengthen the link between the sponsoring institution's educational mission and its public service obligation.

Adult↗

Ultraviolet irradiation produces novel endonuclease III-sensitive cytosine photoproducts at dipyrimidine sites.

Ultraviolet light irradiation of DNA in vitro and in vivo induces cyclobutane dimers, (6-4) pyrimidine-pyrimidone photoproducts and a variety of minor products. Using a defined DNA fragment, we have identified two classes of sites that can be cleaved by Escherichia coli endonuclease III: single cytosines whose heat lability corresponds to that of cytosine hydrates and more heat-stable dipyrimidines containing cytosine. The dipyrimidine products are induced at sites suggestive of (6-4) photoproducts but are not recognized as (6-4) photoproducts by radioimmunoassay. Use of oligonucleotides containing a single cyclobutane thymine dimer, a (6-4) photoproduct or the Dewar photoisomer of the (6-4) photoproduct also indicated that these products are not substrates for endonuclease III. We have therefore identified a minor UV photoproduct that has the same sequence specificity as the two major dipyrimidine photoproducts; it may be a minor isomer, a unique derivative or an oxidative lesion confined to dipyrimidine sites. Its biological significance is not yet known but may be masked by the preponderance of major products at the same sites. Its occurrence at the particular site in dipyrimidine sequences involved in the mutagenic action of UV photoproducts suggests that it may play a role in generating C to T transitions that are common UV-induced mutations.

Animals↗

Preferential selection of receptor-binding variants of influenza virus hemagglutinin by the neutralizing antibody repertoire of transgenic mice expressing a human immunoglobulin mu minigene.

An analysis was made of the neutralizing antibody repertoire, for influenza virus hemagglutinin (HA) of transgenic mice expressing a human immunoglobulin mu (IgH) minigene, by monoclonal antibody (MAb) selection and sequencing of the HA genes of X31 (H3N2 subtype) laboratory variants. Whereas previously reported laboratory variants, selected in ovo with high-affinity murine MAbs of the IgG class, differed from wild-type virus by a single amino acid residue change in one of the major antigenic sites, neutralizing MAbs from transgenic donors selected novel variant viruses with altered receptor-binding specificity and contained residue changes in both the receptor-binding pocket (HA1 225 or HA1 226) and an antigenic site (HA1 135, HA1 145, or HA1 158). Changes in receptor-binding specificities of the variant viruses were confirmed by their resistance to inhibition by horse serum glycoproteins and altered binding to neoglycoproteins. The residue changes in variant virus V-21.2 (HA1 135 G-->R, 225 G-->D) abrogated neutralization by each of the MAbs; nevertheless V-21.2 was recognized by its own selecting MAb in enzyme-linked immunosorbent assay and therefore qualified as an adsorptive mutant rather than an antigenic variant. We consider that a low-affinity neutralizing antibody response may preferentially select for receptor-binding variants of influenza virus HA.

Animals↗

N-acetyltransferase 2 genotype in colorectal cancer and selective gene retention in cancers with chromosome 8p deletions.

BACKGROUND: Genetic polymorphisms in N-acetyltransferase (NAT2) can change the normally fast acetylation of substrates to slow acetylation, and have been associated with the development of some cancers. The NAT2 locus may also suffer dysregulation during cancer progression, as the gene resides on chromosome 8p22, a region which is frequently deleted in colorectal cancer. SUBJECTS AND METHODS: A polymerase chain reaction based method was used to determine NAT2 genotype in 275 patients with colon cancer and 343 normal control DNAs. Within the cancer group, 65 cases known to contain deletions in chromosome 8p were examined for loss of heterozygosity at the NAT2 locus. RESULTS: Overall, there was no statistical difference in frequency or distribution of NAT2 alleles and genotype between colon cancer and control groups. There was a significant association between the slow acetylation genotype and early age of onset. NAT2 genotype did not vary with other clinical features of colon cancer, which included Dukes's stage, site of tumour, and sex. Of 48 informative cases, only three (6%) showed loss of heterozygosity, indicating that the NAT2 locus is not commonly deleted in colorectal cancer. This suggests that NAT2 is retained during the process of allele loss possibly because of its proximity to a gene necessary for cell viability. CONCLUSIONS: NAT2 does not play a major role in colorectal cancer risk, but may influence risk in some age groups. The nature of the loss of heterozygosity at the chromosome 8p site is complex and is worthy of further study.

Acetylation↗

A simplified assay for the arylamine N-acetyltransferase 2 polymorphism validated by phenotyping with isoniazid.

Human arylamine N-acetyltransferase (NAT) activity is determined by two distinct genes, NAT1 and NAT2, and the classical acetylation polymorphism in NAT2 has been associated with a variety of disorders, including lupus erythematosus and arylamine induced cancers. Over 50% of the white population exhibit a slow acetylator phenotype. The genetic basis of the defect has been identified and several DNA based assays are available for genotyping studies. We present here a simplified, rapid PCR based assay for the identification of the major slow acetylator genotypes and validate it using isoniazid as probe drug. This assay was 100% predictive of phenotype. The three genotypes (homozygous mutated, heterozygous, and homozygous rapid) corresponded to a trimodal distribution of Ac-INH/INH metabolic ratios (slow, intermediate, and rapid) without overlapping.

Arylamine N-Acetyltransferase↗

Ventilatory effects of specific carotid body hypocapnia and hypoxia in awake dogs.

Specific carotid body (CB) hypocapnia in the-10-Torr (less than eupneic) range reduced ventilation in the awake and sleeping dog to the same degree as did CB hyperoxia [CB PO2 (PCBO2); > 500 Torr; C.A. Smith, K.W. Saupe, K. S. Henderson, and J. A. Dempsey. J. Appl. Physiol. 79:689-699, 1995], suggesting a powerful inhibitory effect of hypocapnia at the carotid chemosensor over a range of PCO2 encountered commonly in physiological hyperpneas. The primary purpose of this study was to assess the ventilatory effect of CB hypocapnia on the ventilatory response to concomitant CB hypoxia. The secondary purpose was to assess the relative gains of the CB and central chemoreceptors to hypocapnia. In eight awake female dogs the vascularly isolated CB was perfused with hypoxic blood (mild, PCBO2 approximately equal to 50 Torr or severe, PCBO2 approximately equal to 36 Torr) in a background of normocapnia or hypocapnia (10 Torr less than eupneic arterial PCO2) in the perfusate. The systemic (and brain) circulation was normoxic throughout, and arterial PCO2 was not controlled (poikilocapnia). With CB hypocapnia, the peak ventilation (range 19-27 s) in response to hypoxic CB perfusion increased 48% (mild) and 77% (severe) due to increased tidal volume. When CB hypocapnia was present, these increases in ventilation were reduced to 21 and 27%, respectively. With systemic hypocapnia, with the isolated CB maintained normocapnic and hypoxic for > 70 s, the steady-state poikilocapnic ventilatory response (i.e., to systemic hypocapnia alone) decreased 15% (mild CB hypoxia) and 27% (severe CB hypoxia) from the peak response, respectively. We conclude that carotid body hypocapnia can be a major source of inhibitory feedback to respiratory motor output during the hyperventilatory response to hypoxic carotid body stimulation.

Animals↗

Superior laryngeal nerve section alters responses to upper airway distortion in sleeping dogs.

We investigated the effect of superior laryngeal nerve (SLN) section on expiratory time (TE) and genioglossus electromyogram (EMGgg) responses to upper airway (UA) negative pressure (UANP) in sleeping dogs. The same dogs used in a similar intact study (C. A. Harms, C. A., Y.-J. Zeng, C. A. Smith, E. H. Vidruk, and J. A. Dempsey. J. Appl. Physiol. 80: 1528-1539, 1996) were bilaterally SLN sectioned. After recovery, the UA was isolated while the animal breathed through a tracheostomy. Square waves of negative pressure were applied to the UA from below the larynx or from the mask (nares) at end expiration and held until the next inspiratory effort. Section of the SLN increased eupneic respiratory frequency and minute ventilation. Relative to the same dogs before SLN section, sublaryngeal UANP caused less TE prolongation while activation of the genioglossus required less negative pressures. Mask UANP had no effect on TE or EMGgg activity. We conclude that the SLN 1) is not obligatory for the reflex prolongation of TE and activation of EMGgg activity produced by UANP and 2) plays an important role in the maintenance of UA stability and the pattern of breathing in sleeping dogs.

Animals↗