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Biomedical subjects

J L Smith

Publications and source records attributed to J L Smith.

At least 253 records · Page 14Linked to original sources
PubMed1992

Use of paraffin wax embedded bone marrow trephine biopsy specimens as a source of archival DNA.

AIMS: To evaluate the use of DNA extracted from paraffin wax embedded trephine biopsy specimens as a source of archival nucleic acid for Southern hybridisation studies and polymerase chain reaction (PCR) amplification. METHODS: DNA was extracted simultaneously from paraffin wax embedded bone marrow trephine and lymph node biopsy specimens after incubation of tissue sections for one to five days in lysis mix and proteinase K with periodic sampling. DNA from 10 trephine biopsy specimens was subjected to PCR amplification using HLA-DPB primers to determine whether the extracted nucleic acid was of sufficient quality to permit amplification. RESULTS: For most specimens the greatest yield of high molecular weight DNA was seen after five days' incubation. Unlike lymph node material the quality of extracted nucleic acid and the quantity obtained from trephines was insufficient for Southern blot analysis. PCR amplification using HLA-DPB primers yielded positive results in six out of 10 trephine biopsy specimens. CONCLUSIONS: DNA extracted from paraffin wax embedded trephine biopsy specimens is largely degraded and unsuitable for Southern analysis but serves as a useful source of archival nucleic acid for PCR amplification.

Biopsy↗
PubMed1992

Pneumocandins from Zalerion arboricola. III. Structure elucidation.

Pneumocandin B0 (6) and six related lipopeptides are antifungal and anti-Pneumocystis carinii agents from mutants of Zalerion arboricola, whose structures were determined mainly on the basis of spectroscopic analysis. They belong, along with pneumocandin A0 (L-671,329) previously isolated from these laboratories, to the echinocandin class of antifungal agents. The product from base-catalyzed ring opening involving the hemiaminal position of the dihydroxyornithine residue of B0, has been clearly defined as 6b. Modifications were limited to the 3-hydroxy-4-methylproline, 3,4-dihydroxyhomotyrosine and 4,5-dihydroxyornithine residues of pneumocandin A0.

Anti-Bacterial Agents↗
PubMed1991

oskar mRNA is localized to the posterior pole of the Drosophila oocyte.

Mutants of the maternal posterior-group genes of Drosophila lack posterior body pattern elements and germ cells, both of which form through the action of determinants localized to the posterior pole of the oocyte. We report that transcripts of one of these genes, oskar, become localized to the posterior pole of oocytes shortly after the oocyte begins to differentiate visibly. Analysis of various posterior-group mutants reveals that localization of oskar mRNA is an early step in the posterior localization pathway. In addition, we find that nonsense oskar mutations disrupt osk mRNA localization, while missense oskar mutations do not.

Amino Acid Sequence↗
PubMed1991

Malignant blue nevus.

To elucidate the histologic features and biologic behavior of malignant blue nevus (MBN), 12 cases were analyzed in which the tumor showed no junctional activity and arose in a blue nevus background. Seven patients were men and five were women; their mean age was 48.8 years. Eight lesions were on the scalp, and no patient had a family history of melanoma. The histologic appearance of most lesions was a nodule or nodules of malignant cells in a blue nevus. Mitoses were present in all lesions with atypical forms in eight; however the mitotic rate exceeded two per ten high-power fields in only one lesion. Four lesions had necrosis, and four had a heavily pigmented malignant component. Four patients had recurrences; ten patients had metastases, and eight died of their disease. Therefore MBN is an aggressive neoplasm.

Adult↗
PubMed1991

Further evidence of extrinsic forces in bending of the neural plate.

Bending of the neural plate has long been considered to be driven by principally intrinsic forces generated by wedging of neurepithelial cells. Our previous studies have shown that during neural fold elevation, significant neurepithelial cell wedging occurs only within the median hinge point (MHP), the midline region of neural plate anchored to the notochord. We have also shown that neural fold elevation can still occur when MHP cells are prevented from becoming wedge-shaped but fails to occur when the neural plate is separated from lateral nonneurepithelial tissues, even though MHP cells still become wedge-shaped and the midline neural plate still furrows. Together, these results suggest that neural fold elevation, rather than being driven by neurepithelial cell wedging, is driven, at least in part, by extrinsic forces generated by lateral nonneurepithelial tissues. However, it could be argued that in the absence of localized neurepithelial cell wedging, compensatory and atypical cell wedging occurred uniformly throughout the neural plate, providing forces adequate for neural fold elevation. Likewise, it could be argued that in the process of separating the neural plate from lateral nonneurepithelial tissues, the neural plate was damaged to the extent that the neural folds were unable to elevate. To investigate the validity of these arguments, we removed the following tissues microsurgically prior to neural fold elevation: MHP cells, varying amounts of lateral neurepithelial cells (L cells), and the tissues directly underlying these two populations of neurepithelial cells. We found that the neural folds still formed and underwent elevation, convergence, and fusion, resulting in an essentially normal neural tube, even though MHP cells, the underlying notochord, and some L cells were absent for long craniocaudal distances. These results demonstrate that microsurgery alone does not damage the neural plate sufficiently to prevent neural fold elevation, convergence, and fusion. Moreover, the fact that each of the two persisting remnants of lateral neurepithelium generally remained straight and consistently changed their orientation from horizontal to vertical rather than curling suggests very strongly that bending of the neural plate in these embryos is not the result of compensatory and atypical cell wedging. Finally, the results provide further direct evidence of extrinsic forces in bending because the two remnants of lateral neurepithelium, which were oriented horizontally at the time of tissue extirpation, could not have become oriented vertically in the absence of such forces.

Aging↗
PubMed1991

Crystallization and preliminary characterization of mitogillin, a ribosomal ribonuclease from Aspergillus restrictus.

Mitogillin is a ribonuclease secreted by the fungus Aspergillus restrictus. The substrate for mitogillin is a short, universally conserved, sequence in ribosomal RNA. Cleavage of this sequence inactivates protein synthesis. Mitogillin was crystallized by a two-chamber vapor/liquid diffusion method using ethanol as the precipitant. This method has wider potential in the use of volatile organic solvents as precipitants. Crystals of mitogillin diffract X-rays to lattice d-spacings of at least 1.6 A, and belong to the monoclinic space group P2(1), with a = 50.4 A, b = 82.4 A, c = 38.2 A and beta = 99.8 degrees.

Allergens↗
PubMed1991

A prospective study of advance directives for life-sustaining care.

BACKGROUND: The use of advance directives is recommended so that people can determine the medical care they will receive when they are no longer competent, but the effectiveness of such directives is not clear. METHODS: In a prospective study conducted over a two-year period, 126 competent residents of a nursing home and 49 family members of incompetent patients were interviewed to determine their preferences with respect to hospitalization, intensive care, cardiopulmonary resuscitation, artificial ventilation, surgery, and tube feeding in the event of critical illness, terminal illness, or permanent unconsciousness. Advance directives, consisting of signed statements of treatment preferences, were placed in the medical record to assist in care in the nursing home and to be forwarded to the hospital if necessary. RESULTS: In an analysis of 96 outcome events (hospitalization or death in the nursing home), care was consistent with previously expressed wishes 75 percent of the time; however, the presence of the written advance directive in the medical record did not facilitate consistency. Among the 24 events in which inconsistencies occurred, care was provided more aggressively than had been requested in 6 cases, largely because of unanticipated surgery or artificial ventilation, and less aggressively than requested in 18, largely because hospitalization or cardiopulmonary resuscitation was withheld. Inconsistencies were more likely in the nursing home than in the hospital. CONCLUSIONS. The effectiveness of written advance directives is limited by inattention to them and by decisions to place priority on considerations other than the patient's autonomy. Since our study was performed in only one nursing home and one hospital, other studies are necessary to determine the generalizability of our findings.

Adult↗
PubMed1991

Coupled and uncoupled limb oscillations during paw-shake response.

Intersegmental limb dynamics and muscle activities were analyzed for consecutive cycles of paw-shake responses from chronic-spinalized cats to investigate how hindlimb trajectories organize into a pattern with regular oscillations, a steady-state response, or alternatively, into a pattern with irregular oscillations, a nonsteady-state response. In the spinalized preparation, steady-state and nonsteady-state responses have an equal likelihood of emerging from the initial cycles of a paw-shake response, suggesting that regular coupling of joint oscillations is not planned by pattern-generating networks within lumbosacral segments. To examine the characteristics of coupled and uncoupled limb oscillations during paw-shake responses, we assessed patterns of muscle activity and hindlimb kinematics of six adult chronic-spinalized cats. Additionally, we used inverse-dynamics techniques to quantify the intersegmental dynamics of the paw, leg, and thigh. Our data indicate that by the second cycle of both steady-state and nonsteady-state responses, the basic pattern of interaction between muscle and motion-dependent torques at the ankle and knee joints was established. During subsequent cycles of steady-state responses, a consistent sequence of timing changes occurred, such that, just prior to steady-state oscillations, torque maximums peaked simultaneously at each joint and joint reversals occurred simultaneously. Although nonsteady-state responses showed a similar sequence during beginning cycles, increased ankle muscle and net torques during middle cycles created larger inertial torques at the knee joint that were not counteracted and resulted in irregular and uncoupled knee oscillations. It is likely that neither steady-state nor nonsteady-state oscillations are planned by pattern-generating networks within lumbosacral segments, but that patterns of interjoint coordination emerge from the coupling among oscillators. For paw-shake responses in the spinalized preparation, coupling may depend on interactions between central circuits and motion-dependent feedback that is necessary to stabilize inertial effects due to large ankle joint accelerations.

Activity Cycles↗
PubMed1991

Sequence of the cloned pyr4 gene of Trichoderma reesei and its use as a homologous selectable marker for transformation.

We have cloned and sequenced the Trichoderma reesei pyr4 gene encoding orotidine-5'-monophosphate decarboxylase. Comparison of this sequence with that of the equivalent gene from other filamentous fungi suggests that T. reesei is closely related to Cephalosporium acremonium and Neurospora crassa. The cloned pyr4 gene has been used as a homologous selectable marker for transformation of T. reesei. The majority of transformants obtained with circular plasmid were mitotically unstable and contained non-integrated plasmid molecules, sometimes in addition to plasmid integrated in the genome, Linearization of plasmid prior to transformation decreased the transformation frequency but increased the proportion of stable transformation obtained.

Amino Acid Sequence↗
PubMed1991

Psychophysical lifting capacity over extended periods.

The effect of time on an individual's lifting capacity over extended periods using a psychophysical approach was studied. Twelve male subjects estimated their lifting capacity in a 25 min period, and then attempted to lift this weight for an 8 h period under varying conditions. For one experimental condition the subjects were allowed to adjust the weight, the final adjusted maximum acceptable weight of lift (MAWOL) averaged 85.4% of the original MAWOL determined in the 25 min session. The subjects also attempted to lift for an 8 h period, without any weight adjustments. All 12 subjects lasted the 8 h at 2 lifts per min, but at a frequency of 8 lifts per min only three subjects completed the eight hour lifting task. This indicates that the psychophysical approach is a valid method to measure lifting capacity across the lower lifting frequency range but overestimates the lifting capacity at the higher frequency range. Slight fluctuations were noted in heart rate and oxygen consumption which were recorded every hour.

Adult↗
PubMed1991

Urinary levels of activated trypsin in whole-organ pancreas transplant patients with duodenocystostomies.

Urinary levels of trypsinogen and active trypsin were measured in 34 urine samples from 14 patients an average of 23 months following whole organ pancreas transplantation and duodenocystostomy. Timed urine specimens obtained 30 min and 90 min following a time zero void were collected from 5 patients in an effort to define the kinetics of trypsinogen secretion and activation. Total urinary protein and urinary pH were correlated with urinary levels of trypsin and trypsinogen. Twenty-one specimens from 8 normal volunteers and a single specimen from a pancreas transplant patient with a duodenoenterostomy served as controls. Activated trypsin was present in 33 of 34 specimens from 13 of the 14 transplant patients. The average total trypsin activity in all samples was 84.4 micrograms/ml urine (+/- SE 9.6). Trypsinogen was present in 13 of 34 samples, from 7 of 14 patients. The average trypsinogen concentration of all 34 samples was 9.6 +/- 6.2 micrograms/ml. No trypsin or trypsinogen activity was identified in any control sample. In the 5 patients undergoing timed urine collections total trypsin increased an average of 1.3-fold at 30 min and 1.1-fold at 90 min relative to time zero. Urinary trypsinogen increased an average of 7.1-fold at 30 min and 3.1-fold at 90 min following the initial void. Urinary pH and total urinary protein failed to show a significant correlation with urinary levels of total trypsin or trypsinogen. These data suggest that trypsinogen is rapidly converted to active trypsin following secretion into the bladder, resulting in the high urinary trypsin levels that were detected in the majority of patients.

Cystostomy↗
PubMed1991

Variable X-chromosome DNA methylation patterns detected with probe M27 beta in a series of lymphoid and myeloid malignancies.

In this study the X chromosome probe M27 beta was used to investigate DNA methylation at the DXS255 locus and hence X inactivation status and determination of tumour clonality in blood, bone marrow and biopsy tissue involved with morphologically and phenotypically defined lymphoid and myeloid disease from 14 female patients along with uninvolved bone marrow from two control individuals. Thirteen out of 16 individuals (81%) were restriction fragment length polymorphism (RFLP) heterozygous for DXS255. DNA methylation status could not be assessed in the three DXS255 homozygous individuals. In eight DXS255 heterozygous individuals clonality was clearly demonstrated using M27 beta and in six of these cases independent analysis using T cell receptor (TcR) and immunoglobulin (Ig) gene probes confirmed the presence of clonal tumour cell populations. In the two controls, polyclonality was inferred from M27 beta probe analysis. In the remaining three cases (all acute lymphoblastic leukaemia (ALL)) both DXS255 X chromosome sequences appeared to be methylated. Clonality in these cases was demonstrated by TcR or Ig monoclonal gene rearrangements. These data demonstrate the value of the M27 beta probe for determining tumour clonality in a number of cases with lymphoid and myeloid disease but indicate that there may not always be a complete correlation between DNA methylation. X inactivation status and tumour clonality in certain lymphoid neoplasms, restricting the use of this probe in clonality studies. Correlations between DNA methylation, X inactivation status and stage of normal and neoplastic T and B cell development require further investigation.

Antigens, Neoplasm↗