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Biomedical subjects

M M Miller

Publications and source records attributed to M M Miller.

At least 73 records · Page 4Linked to original sources

Characterization of the tissue regression process in the uterus of older mice as apoptotic by the presence of Tp30, an isoform of terminin.

Whether old uteri that have undergone involution do so by an apoptotic mechanism was examined by the presence of known biochemical and morphological markers for programmed cell death. Terminin, a protein identified by an unique monoclonal antibody, has three forms, Tp-90, Tp-60, and Tp-30: Tp-90 (the 90 kDa form) is only present in growing and quiescent non-growing cells; Tp-60 (the 60 kDa form) is found in senescent cells; and finally, Tp-30 (the 30 kDa form) is found in cells committed to apoptotic death. Biochemical analysis of a protein, Tp30, previously identified as a marker for the commitment to programmed cell death, was performed with both young (5-month-old) and old (24-month-old) C57BL/6J mouse uteri. In addition to biochemical analysis of Tp30 presence in uterine tissue, propidium iodide (PI) staining and DNA framentation by nick-end labelling with fluorescence-conjugated UTP were used to characterize apoptosis-related changes in the chromatin organization of the nucleus. Results indicate that within the old uterus Tp-30 is indeed detected in the tissue extracts and was the major terminin band, while Tp-90 and Tp-60 were the major bands observed in extracts of the younger mouse uterus. The presence of Tp30 in the older uterine tissue suggests that the tissue regression which has occurred in the uterus of older mice may be apoptotic in nature. This suggestion is further supported by the demonstration of increases in the number of cells showing apoptotic morphology, i.e. positive staining with UTP reflecting the presence of nuclei with nicked DNA, localized exclusively in the uterine stroma of older females. The presence of DNA fragmentation, as reflected by UTP staining, was virtually absent from the young uteri. These data suggest that apoptosis may be a part of the cellular mechanism contributing to the regression of uterine tissue in the older female during involution, appearing as an age-dependent event.

Aging↗

An Internet-based nuclear medicine teaching file.

UNLABELLED: Teaching file cases play an important role in the training of nuclear medicine residents; however, film-based teaching files have limitations, such as difficulty in accessing cases in a department with several remote clinical sites. The goal of this project was to develop a digital teaching file with the capability for local and remote (Internet) network access, with the additional requirements that viewing existing cases and addition of new cases be easy and simple. METHODS: The teaching file software (TF-Web) utilizes applications developed for the World-Wide-Web in combination with locally developed programs for importing images, entering case information, indexing, searching, case selection and case editing. The time required to add cases to the TF-Web and to access existing cases from local and remote network sites as well as computer storage requirements were assessed. RESULTS: Cases entered in TF-Web may be viewed either with or without diagnoses and may be accessed with acceptable speed (2-14 sec) from both local and remote network sites. A relatively complex case required 1.2 megabytes of storage, with lesser storage requirements for simpler cases. CONCLUSION: A digital teaching file has been developed that allows easy access from computers located both locally and elsewhere on the Internet. Digital storage requirements are reasonable, and, because of the unique nature of TF-Web, case storage may be distributed among multiple institutions.

Computer Communication Networks↗

Two Mhc class I and two Mhc class II genes map to the chicken Rfp-Y system outside the B complex.

Gene sequences highly similar to major histocompatibility complex (Mhc) class I and class II genes were recently recognized as mapping to a site in the genome of the chicken separate from the Mhc class I, class II, and B-G genes of the major histocompatibility (B) complex. The present study was undertaken to see whether this complex of Mhc-like genes designated as restriction fragment pattern Y (Rfp-Y) might reside in one of three clusters of cosmid clones contained within the molecular map of chicken Mhc genes, since only two of the three clusters can be assigned to the B system. To determine whether the third cluster (cluster II/IV) might contain Rfp-Y, a subclone (18.1) from within cluster II/IV near a polymorphic lectin gene was used to analyze the DNA of families in which Rfp-Y haplotypes are known to be segregating. The restriction fragment polymorphisms revealed by the 18.1 probe were found to segregate in parallel with the restriction fragment polymorphisms defining the Rfp-Y haplotypes, thus establishing the location of Rfp-Y within cosmid cluster II/IV. Two of six Mhc class I genes and two of five Mhc class II genes map to cosmid cluster II/IV, so a substantial fraction of chicken Mhc genes, including at least one that may be expressed, are located in a chromosomal region separate from the B system. In further linkage analyses, Rfp-Y was found to assort independently from more than 400 markers in the present linkage map of the chicken genome.

Alleles↗

Syngeneic lysis of reticuloendotheliosis virus-transformed cell lines transfected with Marek's disease virus genes by virus-specific cytotoxic T cells.

Cell-mediated immune responses against Marek's disease virus (MDV) antigens were examined using reticuloendotheliosis virus (REV)-transformed cell lines of two haplotypes (B19B19 and B13B13). These cell lines were stably transfected with cloned fragments of MDV DNA resulting in the expression of the MDV-specific phosphoprotein pp38. Effector cells were obtained from P2a (B19B19) and S13 (B13B13) chickens at 7 days post inoculation with REV, oncogenic or attenuated serotype 1 MDV (JM-16/O and JM-16/A, respectively), serotype 2 MDV (SB-1), or herpesvirus of turkeys (HVT). Transfection of MDV genes did not influence the expression of Class I major histocompatibility complex antigens. The optimal effector to target cell ratio was determined to be 100:1. REV-sensitized effector cells lysed REV cell lines and REV cell lines transfected with MDV DNA in a syngeneic fashion. Effector cells from chickens inoculated with JM-16/O, JM-16/A, SB-1 or HVT lysed only the syngeneic, transfected cell lines, but not the parent REV cell lines. The percentage specific release caused by the MDV-sensitized effector cells was low, but statistically significant.

Animals↗

Current trends in the primary care management of chronic congestive heart failure.

Congestive heart failure affects over 2.3 million Americans; approximately 400,000 new cases are diagnosed yearly in this country. Congestive heart failure is a complex disorder with a poor long-term prognosis. The major causes of congestive heart failure are ischemia, infarction, and idiopathic cardiomyopathy. Patients often present with dyspnea and a low exercise tolerance. In congestive heart failure there may be an alteration in preload, afterload, and contractility of the heart. Many compensatory mechanisms occur to support the failing heart. Cardiac symptoms slowly develop and eventually systemic symptoms develop. The cornerstones of pharmacologic therapy are cardiac glycosides, diuretics, and angiotensin-converting enzyme inhibitors.

Cardiovascular Agents↗

Ontogeny of arginine vasopressin-immunoreactive neurons in the hypothalamus of fetal and newborn sheep.

Arginine vasopressin (AVP) is a peptide hormone which is found in neurons within the paraventricular (PVN) and the supraoptic (SON) nuclei of the hypothalamus. In fetal sheep, this neuropeptide is involved in maturational processes and adaptive responses to 'stress'. This study examined the effect of age on the total number and distribution of AVP-containing neurons in the PVN and SON of fetal sheep and newborn lambs by quantitative light-microscopic immunocytochemistry. Serial coronal sections of hypothalami from three groups of animals were studied: fetuses at 104-109 days of gestation (n = 6) comprising the early group, fetuses at 130-139 days of gestation (n = 5) comprising the late group and newborn lambs at 12-20 postnatal days (n = 5) comprising the neonatal group. This period of development was chosen since adaptive mechanisms to stress are operative at or near the time of birth. Hypothalamic dimensions were measured to determine if maturation had an effect on the size of the AVP-containing subregions of the hypothalamus during this period of development. Dimensions included: ventricle height, optic chiasm width, distances from the dorsal margin of the ventricle to the lateral and medial margins of the optic tract, and distance between the medial margins of the optic tracts. As expected, with increase in maturational age, overall dimensions of the AVP-containing subregions increased significantly (p < 0.05). When early- and late-gestation fetuses were compared to newborn lambs, there was a significant increase in the total number of immunoreactive neurons in both the PVN (p < 0.01, Anova) and SON (p < 0.001, Anova) with age. With advancing age, we also observed an increase in the density of AVP neurons in the middle subregion of the PVN and in the midrostral subregion of the SON. These data suggest that, during the late gestational and early postnatal period, de novo synthesis of AVP genes occurs in these hypothalamic nuclei. This study provides a baseline for further investigation to study the effects of stress on these neurons in the developing ovine fetus and newborn lamb.

Aging↗

Prevalence of spondyloarthropathies in Alaskan Eskimos.

OBJECTIVE: To estimate the prevalence of spondyloarthropathies (SpA) in 2 Alaskan Eskimo populations, using improved methodology for case ascertainment and new, more inclusive classification criteria. METHODS: Through existing rheumatic disease registries, health care providers and the Alaska Area Native Health Service (AANHS) computerized patient information system, we identified all native residents of the 2 study regions with a diagnosis of any inflammatory arthritis or problems characteristic of SpA, such as iritis or persistent back pain. Individuals with such diagnoses or problems were evaluated in clinic, according to a standardized protocol (interview, examination), and by medical record review, pelvic radiography and laboratory tests. Each case was evaluated according to standard diagnostic criteria for the individual disease entities and by the Amor and European Spondylarthropathy Study Group (ESSG) preliminary classification criteria for SpA. RESULTS: We identified 104 cases of SpA in the combined Eskimo populations, an overall prevalence of 2.5% in adults aged 20 and over. Both undifferentiated (USpA) and reactive SpA were more common than ankylosing spondylitis (AS). CONCLUSION: Using the new criteria and a more effective approach to case ascertainment we found the prevalence of SpA to be about twice that found in our earlier studies of adult Eskimo populations. The prevalence of SpA was very similar in men and women. Despite the known high prevalence (25-40%) of HLA-B27 in the study populations we did not find the prevalence of any form of SpA to be as strikingly high as that of AS (6-10%) for the Canadian Haida.

Adolescent↗

Modulation of a renal estrogen receptor by protein kinase C.

In this paper we demonstrate the existence of specific, high affinity 17-beta estradiol binding in MDBK cells (a normal non-transformed renal cell line). Scatchard analysis revealed binding characteristics typical of the estrogen receptor (ER). Only unlabelled 17-beta estradiol, diethyl stilbestrol and estrone effectively competed with [3H] 17-beta estradiol for binding, other steroids did not compete. Short term TPA treatment of MDBK cells with TPA increased PKC activity and immunoreactivity and caused a transient increase in 17-beta estradiol binding, while longer term treatment with TPA decreased PKC activity and immunoreactivity. The inactive phorbol ester 4 alpha PDD was without effect on PKC activity and the ER. TPA did not affect the affinity of the ER for the nucleus nor did it increase degradation of the receptor. We hypothesize that the renal ER may be a substrate for PKC and its properties can be modified by PKC similarly to the vitamin D receptor.

Animals↗

Beta IV is the major beta-tubulin isotype in bovine cilia.

Four different isotypes of beta-tubulin are known to be expressed in mammalian brain. Monoclonal antibodies against beta II, beta III, and beta IV were used to characterize the beta-tubulin isotypes in two ciliated bovine tissues: non-motile sensory cilia of retinal rod cells and motile cilia of tracheal epithelium. Retinal rod outer segment (ROS) connecting cilia and cytoskeletons were purified by density gradient centrifugation. This preparation contained more than 20 major protein components, as shown by dodecyl sulfate polyacrylamide gel electrophoresis. Electroblots were used to quantitate the relative amounts of beta II, beta III, and beta IV. The connecting cilium and cytoskeleton of the rod outer segment has less type III beta-tubulin than brain and more type IV. The ratio of beta IV to beta II in the ROS is nearly a factor of 8 larger than in brain. Electron microscopic immunocytochemistry showed extensive labeling of cilia by anti-type IV in thin sections of retinas and trachea, and also in purified ROS cilia and cytoskeletons. Labeling of cilia by anti-beta II was also observed, although in the purified ROS cilia and cytoskeleton, the anti-beta II labeling was primarily on amorphous non-ciliary material. The results suggest that both motile and non-motile cilia are enriched in the type IV beta-tubulin subunit.

Amino Acid Sequence↗

A polymorphic system related to but genetically independent of the chicken major histocompatibility complex.

Analyses of the major histocompatibility complex (Mhc) in chickens have shown inconsistencies between serologically defined haplotypes and haplotypes defined by the restriction fragment patterns of Mhc class I and class II genes in Southern hybridizations. Often more than one pattern of restriction fragments for Mhc class I and/or class II genes has been found among DNA samples collected from birds homozygous for a single serologically defined B haplotype. Such findings have been interpreted as evidence for variability within the Mhc haplotypes of chickens not detected previously with serological methods. In this study of a fully pedigreed family over three generations, the heterogeneity observed in restriction fragment patterns was found to be the result of the presence of a second, independently segregating polymorphic Mhc-like locus, designated Rfp-Y. Three alleles (haplotypes) are identified in this new system.

Animals↗

Modulation of hypothalamic mu-opioid receptor density by estrogen: a quantitative autoradiographic study of the female C57BL/6J mouse.

The labelling of hypothalamic binding sites by [125I]-FK, a specific mu-opioid receptor ligand, was studied in female C57BL/6J mice to test whether removal of ovarian steroids affected the density of distribution of receptor binding. Labelling densities in the forebrain of normally cycling (intact) females (N = 12), were compared to those in mice that had been ovariectomized (OVX) for 6 weeks (n = 8) and in mice that had been OVX and implanted with an estradiol (E2) capsule (OVX+E2) for 6 weeks (n = 11). Frozen sections from the rostral forebrain were incubated with 1 nM [125I]-FK and processed for light microscopic autoradiography. The diagonal band of Broca (DBB), organum vasculosum lamina terminalis (OVLT), preoptic area (POA), septum, parietal cortex, and striatum were analyzed using computerized image analysis. The distribution of labelling was similar in all three experimental groups in all the regions; however, labelling was significantly reduced in the ventrolateral POA of OVX animals compared to intact females. Labelling densities in the OVX animals replaced with the gonadal steroid estradiol were not significantly different from those in normally cycling mice. This study demonstrates a region-specific loss of mu-opiate receptor labelling following long-term deprivation of gonadal steroids, and supports the hypothesis that estrogen directly or indirectly influences the density of mu-opioid receptors in the rostral forebrain of female mice.

Animals↗

Gonadotropin hormone-releasing hormone induced luteinizing hormone responses in young and old female C57BL/6J mice.

The estrogen-induced luteinizing hormone (LH) surge is markedly attenuated in old (24 months) female mice compared with young (6 months) females. To test whether or not this attenuated LH response is due to a diminished capacity of the pituitary to respond to hypothalamic gonadotropin hormone-releasing hormone (GnRH), the pituitary response to exogenous GnRH was measured in young (5-6 months), normally cycling (n = 6) and old (24 months), acyclic constant diestrus (n = 6) C57BL/6J female mice. Mice were ovariectomized and estrogen-treated for 7 days. After intrajugular catheterization on Day 6, serial blood samples were taken at 15-min intervals for 165 min on Day 7. Serum LH was measured in samples obtained before and after infusion of either saline or GnRH (5 micrograms/5 microliters saline/kg body wt) and 15 micrograms/15 microliters saline/kg body wt 1 hr apart). Saline-treated animals demonstrated no LH response in either young (0.09 +/- 0.02 ng/ml baseline) or old (0.11 +/- 0.01 ng/ml baseline) females. However, a significant release of LH was obtained in response to each challenge of GnRH in both young (0.3 +/- 0.04 ng/ml first challenge, 0.69 +/- 0.1 ng/ml second challenge) and old (0.78 +/- 0.1 ng/ml first challenge, 1.76 +/- 0.2 ng/ml second challenge) mice. The LH response in the aged group was significantly greater (P < 0.05, analysis of variance) than in the young group. These results show that pituitaries of old female mice were at least as capable of responding to exogenous stimulation by GnRH as those of young. We conclude that alteration in the capacity of the pituitary to respond to GnRH is not likely to be a factor contributing to altered LH secretion with age in C57BL/6J mice.

Aging↗

Serial transvaginal ultrasound scans and beta-human chorionic gonadotropin levels in early singleton and multiple pregnancies.

OBJECTIVE: To determine if serum beta-hCG levels are higher in multiple gestation than in singleton pregnancy at the time of intrauterine sac visualization and the first appearance of fetal heart activity as documented by serial transvaginal ultrasound (US). DESIGN: Prospective analysis of serial transvaginal US findings in 19 pregnancies correlated with serum hCG levels during early gestation. SETTING: Reproductive endocrinology division of the University of Arkansas for Medical Sciences, Little Rock, Arkansas. PATIENTS: Nineteen infertility patients were studied after conceiving. Thirteen underwent IVF or GIFT, 4 received hMG therapy, 1 was treated with clomiphene citrate, and 1 pregnancy followed spontaneous ovulation. INTERVENTIONS: Transvaginal US and hCG levels were obtained every Monday, Wednesday, and Friday from 20 to 22 days after ovulation until the appearance of fetal heart activity. RESULTS: Initial sac visualization occurred at lower serum hCG levels in singleton versus multiple pregnancies (2,180 +/- 1,170 versus 7,028 +/- 4,280 mIU/mL, mean +/- SD). Sacs were always seen when the serum hCG level (mIU/mL) was > or = 1,161 in singleton, 1,556 in twin, 3,372 in triplet, and 9,399 in quadruplet pregnancies. CONCLUSION: Failure to observe an intrauterine sac by transvaginal US in the presence of serum hCG levels in the 1,000 to 2,000 mIU/mL range is not pathognomonic for an ectopic gestation. Clinical symptomatology, risk of multiple pregnancies, and gestational age must also be considered.

Analysis of Variance↗

The effect of benzo(a)pyrene on murine ovarian and corpora lutea volumes.

OBJECTIVE: Women who smoke have impaired fertility and experience menopause at an earlier age. This experiment determined the effect of benzo(a)pyrene, a polycyclic aromatic hydrocarbon contained in cigarette smoke, on murine ovarian volume, total corpora lutea volume, individual corpora lutea volumes, and corpora lutea numbers. STUDY DESIGN: C57BL/6N mice were treated with intraperitoneal injections of 0 to 500 mg/kg benzo(a)pyrene in corn oil. The 20 mice at each dose were divided into four groups of five each and were killed at 1, 2, 3, or 4 weeks after treatment. Ovaries were serially sectioned and analyzed morphometrically. RESULTS: Benzo(a)pyrene produced a dose- and time-dependent decrease in ovarian volume, total corpora lutea volume, and number of corpora lutea per ovary. This effect was transitory at low doses with complete recovery of corpora lutea by 4 weeks. Compensatory hypertrophy of the individual corpora lutea occurred during the recovery phase. Ovarian function did not return in animals treated with the two highest doses. CONCLUSION: Benzo(a)pyrene is a murine ovarian toxicant that inhibits corpus luteum formation in a dose- and time-dependent fashion.

Animals↗