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Biomedical subjects

D A Miller

Publications and source records attributed to D A Miller.

At least 91 records · Page 5Linked to original sources

Isolation of Leptospira interrogans serovar grippotyphosa from the skin of a dog.

Leptospira interrogans serovar grippotyphosa was isolated from the skin of a 14-year-old male dog with deteriorating health. Necropsy revealed numerous lesions characteristic of aged dogs, but no evidence of acute hepatitis or nephritis, which are common features of pathogenic Leptospira infections. Antibody to Leptospira was not detected in the dog's serum by microagglutination. Leptospires grew slowly in Barbour-Stoenner-Kelly medium, a medium commonly used to isolate Borrelia, but then grew abundantly in Tween 80-bovine albumin leptospire medium. The isolate was pathogenic to a hamster and was identified by microagglutination and restriction endonuclease analysis.

Animals↗

Family with 22-derived marker chromosome and late-onset dementia of the Alzheimer type: II. Further cytogenetic analysis of the marker and characterization of the high-level repeat sequences using fluorescence in situ hybridization.

We have further characterized an unusual 22p+ marker chromosome with a double nucleolus organizer region (dNOR) previously identified in a family with late-onset dementia of the Alzheimer type. G-banding and morphology of the marker's q arm were typically normal. However, the p+ arm had a terminal cytological satellite and a GT-positive region at the midpoint. Standard C-banding documented 2 C-positive regions: one was associated with the primary centromere; the other, which was at the midpoint of the p arm, was not associated with a constriction. With replication-banding, there was a darkly staining region in the middle of the p+ arm that resembled the pericentromeric region of a chromosome 21 or 22. Fluorescence in situ hybridization with pXlr 101, a probe recognizing the full repeating unit of rDNA, indicated that the marker had an unusually larger rDNA region; with pU 1.2, a probe recognizing the human rDNA promoter, the signal was a doublet. The marker had 2 signals with a beta-satellite probe, and a second signal in addition to that present at the primary centromere under low stringency with alpha-satellite probes and a classic satellite probe. Immunostaining of chromosome spreads after R-banding and ultraviolet (UV) denaturation showed that the major portion of the marker's p arm was highly methylated.

Aged↗

Critical thinking in preclinical course examinations.

PURPOSE: To examine the relationship between preclinical medical school course examinations and critical-thinking skills by correlating examination results with the Watson-Glaser Critical Thinking Appraisal (WGCTA), an 80-item critical-thinking inventory. METHOD: The 196 students in the class of 1993 at the University of Texas Southwestern Medical Center at Dallas Southwestern Medical School were given the WGCTA during the second half of their freshman year. A composite WGCTA score and five subtest scores were calculated for each student. The course examinations were 25 tests, each with a majority of multiple-choice items, from courses given during the first two years of the school curriculum. The students' undergraduate grade-point average (GPAs), medical school GPAs, and Medical College Admission Test (MCAT) scores were also included in the analysis. Pearson product-moment correlations were calculated between the students' WGCTA scores and their examination scores, MCAT scores, and GPAs. RESULTS: Sixteen of the examinations--in the behavioral sciences, psychiatry, ethics, pathology, introduction to clinical medicine, genetics, endocrinology, and cell biology--had significant positive correlations with the WGCTA, as did MCAT scores and first-year GPAs. Correlations were more robust with the WGCTA subscales for interpretation (18 examinations), evaluation of arguments (15), and deduction (13), and less robust with the subscales for inference (7) and recognition of assumptions (3). CONCLUSIONS: The results suggest that objective multiple-choice examinations can at least partially reflect critical-thinking skills.

Educational Measurement↗

Targeted brain biopsy: a comparison of freehand computed tomography-guided and stereotactic techniques.

With computed tomography (CT) and magnetic resonance imaging stereotactic systems, biopsies of intracranial lesions can be made with safety and ease. Before the development of this technique, neurosurgeons often performed freehand brain biopsies under CT guidance. While stereotactic biopsy is the procedure of choice for small, deep lesions, few studies have compared the morbidity, mortality, and efficacy in obtaining a diagnosis associated with these two techniques for superficial lesions. A total of 167 consecutive CT-guided or stereotactic brain biopsies were performed in 154 patients. Fourteen of the stereotactic and 12 of the CT-guided biopsies were of deep lesions and were excluded from analysis. The results of 75 freehand CT-guided biopsies of superficial lesions in 69 patients were compared with those of 66 stereotactic biopsies (34 CT-guided and 32 MRI-guided) performed with the Brown-Roberts-Wells stereotactic system in 60 patients. Twenty-five of the lesions in the stereotactic biopsy group measured < or = 2 cm, as compared with 13 of those in the freehand CT-guided biopsy group. There were no biopsy-related deaths among the patients who underwent freehand CT-guided biopsy and one death among those who underwent stereotactic biopsy (1.5%). Freehand CT-guided biopsy was associated with 5% morbidity, compared with 6% morbidity for stereotactic biopsy. Seven CT-guided needle biopsies (9%) and 12 stereotactic biopsies (18%) were nondiagnostic. Statistical analysis showed no significant difference between morbidity and mortality in the two groups, but the rate of diagnostic failure was significant (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Correlates of bulimia nervosa: early family mealtime experiences.

The relationship of early mealtime experiences to later bulimia was investigated in a sample of 128 female college students. Significant group differences were found between bulimics, nonbulimics, and repeat dieters on an early meal experience questionnaire, with the bulimic group reporting the most negative and unusual experiences. Significant differences were also found between groups on depression but not on family adaptability or cohesion. These findings suggest that bulimic, repeat dieter, and nonbulimic females have differentiating histories regarding eating-related issues in the family context.

Adolescent↗

Effect of KTP laser ablation of endochondral bone on bone healing.

The advantages of using a laser to cut a hole in the ossicular chain include increased precision, decreased likelihood of disarticulation, and elimination of noise-induced threshold shift associated with conventional drills. This study was designed to determine optimal energy settings and cooling technique to minimize tissue injury. Laser ablated holes were cut in rabbit femurs with different endostat fibers at power settings of 1.6 to 5.0 watts. Comparable burr holes were cut by a high-speed drill and the animals were sacrificed at different time intervals. Specimens were evaluated microscopically for tissue injury by the amount of cell necrosis of the surrounding osteocytes, bone marrow changes, and denaturation of bone matrix. Changes in healing were assessed by the production and ossification of the granulation tissue. Laser ablated holes created with 1.6 to 2.0 watts at 2 to 7 days demonstrated thermal damage comparable to conventionally drilled holes. At 14 to 28 days these holes demonstrated advanced healing and integrated lamellar bone. In contrast, higher power (> 3.5 watts) at 2 to 7 days demonstrated Significantly more osteocyte loss, marked bone marrow fibrosis, and bone matrix denaturation. At 14 to 28 days these holes showed evidence of delayed healing and osseous plugs not integrated with the surrounding bone.

Animals↗

Pharmacy technicians and computer technology to support clinical pharmacy services.

Use of pharmacy technicians, computer-assisted patient-selection algorithms, and bar-code technology to increase efficiency of clinical pharmacy staff is described. Workload assessment showed that clinical pharmacists at Henry Ford Hospital (Detroit) were spending two thirds of their time with data collection, patient selection, and documentation and only one third of their time intervening to improve drug therapy. To increase pharmacist efficiency, two technicians were hired to perform the clerical functions associated with clinical pharmacy services. Also, a bar-code data-collection system was implemented to help pharmacists document their clinical activity. Computerized algorithms helped pharmacists target patients whose therapy should be most carefully reviewed. Reports generated by the system summarized workload distribution and provided information about the types of problems identified and corrected. After these changes were implemented, the fraction of time that pharmacists spent intervening to improve drug therapy increased to one half. Use of technicians to perform clerical tasks, computerized algorithms to help identify patients in need of pharmacist assistance, and bar-code technology to facilitate data collection has helped clinical pharmacists improve their time management.

Algorithms↗

Characterization of a bifunctional peptidylglycine alpha-amidating enzyme expressed in Chinese hamster ovary cells.

Peptidylglycine alpha-amidating enzyme (alpha-AE) catalyzes the conversion of glycine-extended prohormones to their biologically active alpha-amidated forms. We have derived a clonal Chinese hamster ovary cell line that secretes significant quantities of active alpha-AE. Enzyme production was increased by selection for methotrexate-resistant cells expressing a dicistronic message. Amplification of the alpha-AE gene was monitored by Southern blot analysis, enzyme activity, and immunoreactive protein throughout the selection process. The soluble enzyme is bifunctional as determined by the ability to convert either the glycine-extended substrate, dansyl-Tyr--Val--Gly, or the intermediate, dansyl-Tyr--Val--alpha-hydroxyglycine, to the dansyl-Tyr--Val--NH2 product. The recombinant alpha-AE was purified by a simple two-step chromatographic process. The purified enzyme is partially glycosylated and the glycosylated and nonglycosylated forms of the enzyme were separated on a Con A-Sepharose column. The kinetic constants for dansyl-Tyr--Val--Gly, dansyl-Tyr--Val--alpha-hydroxyglycine, ascorbate, and catechol were the same for both forms of alpha-AE. In addition, mimosine is competitive vs ascorbate with K(is) = 3.5 microM for the nonglycosylated alpha-AE and K(is) = 4.2 microM for the glycosylated alpha-AE. Therefore, the presence or absence of asparagine-linked oligosaccharide does not affect the catalytic efficiency of the enzyme. Overexpression of the recombinant enzyme in CHO cells greatly enhances expression of the endogenous gene, implicating a feedback mechanism on the alpha-AE gene.

Amino Acid Sequence↗

Ullrich-Turner syndrome with a small ring X chromosome and presence of mental retardation.

Since some patients with Ullrich-Turner syndrome (UTS) have mental retardation, we reviewed our experience to look for a high-risk subgroup. Among 190 UTS and gonadal dysgenesis patients with X chromosome abnormalities, 12 had mental retardation. All of the six (100%) with a small ring X were educable (EMI) or trainable mentally impaired (TMI) with more severe delay than expected in UTS. Among the 184 with other X abnormalities, only 6 had similar delays (2 from postnatal catastrophes), for a frequency of 3.3% mental retardation among those without a small ring X; only 2.2% of these had unexplained mental retardation. Polymerase chain reaction studies showed no Y-derived material in the 2 patients who were evaluated, and in situ hybridization confirmed X origin of the ring in the 6 subjects who were evaluated. We describe the phenotype of the 6 individuals with a small ring X, and an additional 2 patients with a small ring X who were identified outside the survey. The subjects with a small ring X comprised a clinically distinct subgroup which had EMI/TMI and shorter stature than expected in UTS. Seizures and a head circumference less than 10th centile were observed in half of the patients with a small ring X, and strabismus, epicanthus, and single palmar creases were present in more than half. A "triangular" face in childhood, pigmentary dysplasia, sacral dimple, and heart defects were also common. Neck webbing appeared to be less frequent than in 45,X. We hypothesize that the high risk of mental retardation in this form of the UTS results from lack of lyonization of the ring X due to loss of the X inactivation center. Excluding those with a small ring X, mental retardation is not significantly increased in patients with UTS.

Adolescent↗

Leptospira serology in small ruminants on St. Croix, U.S. Virgin Islands.

A serological survey of 16 serovars of Leptospira interrogans, previously reported in tropical small ruminants, was undertaken to determine the serovars involved and the prevalence of these antibodies in sheep and goats on St. Croix, U. S. Virgin Islands (USVI). Seven of eight goat herds (108 animals) had at least two seropositive animals in each herd with an individual animal seroprevalence of 26%. The 53 sheep tested (one flock only) showed a 32% seroprevalence. Antibodies against seven serovars were detected in goats (autumnalis, ballum, bataviae, bratislava, canicola, icterohemorrhagiae, and pyrogenes). In addition, hardjo antibodies were detected in sheep. Serovar autumnalis accounted for about 30% of seropositive animals in each species. Many animals showed titers against more than one serovar. The number of seropositive animals suggests Leptospira may be a factor in the health of small ruminants on St. Croix.

Agglutination Tests↗

Methylation of the 5' flanking sequences of the ribosomal DNA in human cell lines and in a human-hamster hybrid cell line.

In a human lymphoblastoid cell line (Z83) in which rDNA genes on chromosome 22 are amplified but transcribed at a low level, immunocytological studies with antibodies to 5 methylcytidine provided evidence for hypermethylation of the rDNA. The extent of methylation of the 5' flanking sequences of the ribosomal DNA was examined by comparing the size of restriction fragments obtained by digestion of genomic DNA with EcoRI and HpaII or EcoRI and MspI. Southern blots indicated hypermethylation of the 5' flanking sequences of many copies of rRNA genes in these cells, but not in a control lymphoblastoid cell line without rDNA amplification. Results obtained with a somatic hybrid human-hamster cell line, in which the rRNA genes on the single human chromosome 22 are inactive, showed that only a small fraction of the CCGG sites in the 5' flanking sequences of the transcriptionally silent rRNA genes in this hybrid were methylated. Since inactive rRNA genes can show such a minimal level of methylation, it is likely that the extreme hypermethylation of the amplified rRNA genes in Z83 occurred in association with their inactivation rather than following it.

Animals↗

Isolation and characterization of a mouse subtelomeric sequence.

A mouse subtelomeric sequence, ST1, was generated from genomic DNA of the mouse HR9 (129/Sv origin) cell line by the polymerase chain reaction (PCR) using a single telomeric primer. ST1 was cloned and characterized: it is composed of 670 bp of novel DNA sequence flanked on each end by inverted telomeric hexanucleotide repeats (TTAGGG)n. PCR amplification from BALB/c mouse DNA using this single primer gave the same major product. Southern analysis and PCR using internal ST1 primers confirmed that the ST1 sequence is present in mouse genomic DNA. In situ hybridization to metaphase chromosomes of SJL origin mapped ST1 to many, if not every, mouse telomere. PCR experiments using different combinations of the telomeric, minor satellite, and ST1 primers indicated that some ST1 copies are adjacent to minor satellite sequences, that telomeric and ST1 sequences are not generally interspersed with minor satellite sequences, and that ST1 and the minor satellite have a consistent and specific orientation relative to each other and to the telomere.

Animals↗

Cross sections of natSb(p,x) reactions for 30-46 MeV protons.

In order to optimize the production of 118Te in thick targets for use in a 118Te/118Sb radionuclide generator, the excitation function for the 121Sb(p,4n)118Te reaction has been measured for 30-46 MeV protons. The excitation functions for the competing reactions natSb(p,xn)119mTe, natSb(p,xn)119Te, natSb(p,xn)121mTe, natSb(p,pxn)120mSb and 123Sb(p,pn)122Sb have also been determined using stacked foil techniques. The 121Sb(p,4n)118Te reaction cross section maximum was found to be 480 mbarn at 44 MeV. In order to minimize the 119m + 119Te interference a minimum proton beam energy of 40 MeV is required. The cross section results are compared with published data and with calculated excitation functions.

Antimony↗

Isolation and comparative mapping of a human chromosome 20-specific alpha-satellite DNA clone.

We have isolated and characterized a human genomic DNA clone (PZ20, locus D20Z2) that identifies, under high-stringency hybridization conditions, an alphoid DNA subset specific for chromosome 20. The specificity was determined using fluorescence in situ hybridization. Sequence analysis confirmed our previously reported data on the great similarity between the chromosome 20 and chromosome 2 alphoid subsets. Comparative mapping of pZ20 on chimpanzee and gorilla chromosomes, also performed under high-stringency conditions, indicates that the alphoid subset has ancestral sequences on chimpanzee chromosome 11 and gorilla chromosome 19. However, no hybridization was observed to chromosomes 21 in the great apes, the homolog of human chromosome 20.

Animals↗

Phenotypic alterations in fibroblasts and fibrosarcoma cells that overexpress latent transforming growth factor-beta 1.

Mouse embryo-derived AKR-2B fibroblasts and murine fibrosarcoma cells (the 1591 cell line) were transfected with a murine transforming growth factor-beta 1 (TGF beta 1) cDNA under the transcriptional control of either the simian virus-40 early promoter or the cytomegalovirus promoter/enhancer. Selected clones secreted 2- to 4-fold more TGF beta-competing activity into their media than the parental cell line or neomycin-transfected controls. The TGF beta 1 released into the cell-conditioned medium was latent. Despite the latency of the overexpressed TGF beta 1, TGF beta 1-transfected cells exhibited phenotypic features of TGF beta 1-treated cells. When confluent, the TGF beta 1-transfected cells had the morphological characteristics of the parental cells that have been treated with active TGF beta 1. AKR-2B cells that expressed higher levels of TGF beta 1 also expressed high levels of c-sis and c-myc mRNAs and decreased TGF beta 2 and TGF beta 3 mRNAs in the same manner as parental AKR-2B cells that had been treated with active TGF beta 1. The transfected 1591 cells that overexpressed TGF beta 1 bound less [125I]TGF beta 1 than did parental 1591 cells, but after a mild acid wash demonstrated an increase in [125I]TGF beta 1 binding. Our results suggest that these TGF beta 1-transfected fibroblast and fibrosarcoma cells have the capacity to activate TGF beta; however, as very little activated TGF beta is detected in the medium, it is hypothesized that these cells activate latent TGF beta 1 and bind the activated TGF beta 1, thus acquiring a phenotype consistent with TGF beta 1-treated cells.

Animals↗

[Chronic gastritis from the aspect of a thrombohemorrhagic syndrome].

As many as 148 patients were examined to demonstrate that chronic gastritis (CG) runs its course in the presence of marked changes in microcirculation and hemostasis which go into the framework of the chronic recurrent thrombohemorrhagic syndrome. The manifestations of the latter one were more appreciable in a dramatic exacerbation of the disease, erosive gastritis, diffuse atrophy of the mucous membrane and secretory insufficiency of the stomach, and disease standing over 10 years. The disorders indicated are involved into the pathogenesis of CG and promote the development of morphofunctional alterations in the gastric mucosa.

Adult↗

Telomeric associations and consistent growth factor overexpression detected in giant cell tumor of bone.

Tumor specimens from 15 patients with giant cell tumor (GCT) of bone were cytogenetically analyzed. A subset of five individuals had tumor cells harvested and polyadenylated RNA isolated. Multiple Northern blots were performed utilizing radiolabeled probes for the growth factors TGF beta 1, TGF beta 2, TGF beta 3, and TGF alpha (TGF, transforming growth factor). RNAs from other types of neoplasms and nonneoplastic cells were examined as controls. The most consistent cytogenetic abnormality detected involved multiple telomeric associations (TAs), most frequently involving the terminus of the long arm of chromosome 19 (19q). Northern blot analysis revealed a consistent expression of TGF beta 1 and TGF beta 2 with an inconsistent mRNA expression for the other TGFs. There was a relative overexpression of mRNA for TGF beta 2. The gene location for TGF beta 1 is near the 19q terminus and thus it is speculated that TGF beta may play a role in the neoplastic transformation of GCT.

Adult↗