Normal symphysis pubis width in children.
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Adolescent↗
Biomedical subjects
K Patel
Publications and source records attributed to K Patel.
Diagnosis of Ewing's sarcoma and peripheral neuroectodermal tumour based on the detection of t(11;22) using fluorescence in situ hybridisation.
Fluorescence in situ hybridisation (FISH) has been used increasingly for gene mapping and ordering probes on interphase and metaphase preparations. The association of consistent chromosomal aberrations with certain malignancies allows the possibility of using interphase cytogenetics as a diagnostic tool. In small round cell tumours of children accurate diagnosis may be difficult using existing methods. We have therefore evaluated the diagnostic potential of this technique when applied to the characteristic t(11;22) found in Ewing's sarcoma and peripheral neuroectodermal tumour (ES and PNET). Interphase nuclei were prepared from normal human foreskin fibroblasts (HFF), two Ewing's sarcoma cell lines and several fresh tumour biopsies. DNA probes each side of the breakpoint at 22q12 were labelled with biotin and digoxygenin, hybridised to chromosomes in interphase and detected in different colours. Measurements between pairs of signals arising from each copy of chromosome 22 were taken and statistical analysis performed. There was a highly significant difference (P < 0.0001) between the two populations of measurements obtained (from nuclei with and without the t(11;22)). Studying four tumours and one further ES line (blind) it was found that median values from 30 nuclei could correctly identify which samples contained the t(11;22). This application of interphase cytogenetics contributes a reliable, accurate and conceptually simple diagnostic test for ES and PNET. It may now be applied to other tumours with characteristic translocations, amplifications or deletions when suitable probes are available. This approach is likely to become a routine in clinical diagnosis.
Chromosomes, Human, Pair 11↗
A disease-associated germline deletion maps the type 2 neurofibromatosis (NF2) gene between the Ewing sarcoma region and the leukaemia inhibitory factor locus.
RFLP typing of members of a neurofibromatosis type 2 (NF2) family suggested that affected individuals were hemizygous at the neurofilament heavy chain (NEFH) locus, possibly as a result of a disease-associated deletion. Conventional karyotyping revealed no evidence for a deletion and all or a majority of the affected family members were heterozygous for closely linked markers which mapped proximal to the NEFH locus (D22S1 and D22S56) and for the distal marker D22S32. FISH analysis confirmed a disease-associated germinal deletion on 22q which encompassed the NEFH locus, which is known to be very closely linked to NF2, but did not extend as far as the proximal Ewing sarcoma region or the distal leukaemia factor (LIF) locus. PFGE analysis with a LIF cosmid subclone identified patient-specific NotI and MluI fragments and suggested that the deletion is about 700 kb in length. Although this large deletion could be expected to eliminate a considerable fraction, and possibly all of the NF2 gene, the resulting phenotype is the mild, so-called Gardner subtype of NF2. The deletion should provide a useful mapping resource for characterising the chromosomal region containing the NF2 locus.
Alleles↗
Impact of treating involuntarily admitted schizophrenics on an open unit.
When involuntary psychiatric patients are treated on unlocked units, problems may arise as to resource expenditure and the ability to provide optimal psychiatric care. This study investigates the problem and possible solutions.
Commitment of Persons with Psychiatric Disorders↗
ACP Broadsheet No 138: May 1993. Gross examination of uterine specimens.
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Biopsy↗
Ordering of probes surrounding the Ewing's sarcoma breakpoint on chromosome 22 using fluorescent in situ hybridization to interphase nuclei.
Eight probes were localized by fluorescent in situ hybridization to the region surrounding the Ewing's sarcoma breakpoint on chromosome 22. Three of these were initially ordered by pair-wise hybridization to metaphase chromosomes with differential detection of the probes. These and the remaining probes were then ordered by hybridizing two or three probes simultaneously to interphase nuclei. In the two probe experiments and some of the three probe experiments, the order was derived by comparing mean interphase distances between signals from the probes. In the three probe experiments, either two probes were detected with one fluorochrome and the third with another or all three probes were individually distinguished by detecting one probe with fluorescein isothiocyanate (FITC), one with Texas Red, and one with both fluorochromes to give a mixed color. The order of the signals was then noted. Greater than 60 percent of configurations with a discrete order were shown or deduced to be correct. These approaches are assessed and we demonstrate a more obvious predominating order when all three probes are differentially detected. The order of the probes was deduced to be centromere: D22S271: D22S260: lambda S1: D22S262: cosK1831: Ewing's sarcoma breakpoint: cosLIF: D22S261: lambda S15: telomere.
Cell Nucleus↗
Detection of significant abnormalities on lumbar spine radiographs.
The purpose of this study was to determine the proportion of significant abnormalities detected on anteroposterior (AP) and lateral radiographs of the lumbar spine when viewed separately, in a series of cases where the prevalence of abnormalities had been artificially increased. Five radiologists of varying experience were required to report separately on the AP and lateral films of 300 cases in which randomly included were 30 cases with metastatic disease, a disc infection or an inflammatory spondylitis. At a later date, unaware of their initial observations, the radiologists repeated the exercise reviewing all the films together. As might be expected the false positive rate was relatively high, particularly in the least experienced radiologists' responses. Nevertheless the overall results indicate that the majority of early inflammatory spondylitis cases will be missed on a solitary lateral film as will many of the metastases. Conversely, fractionally more of the disc infections were observed on the lateral film than on the AP. Further analyses in terms of sensitivity, specificity, positive and negative predictive values are presented. The authors conclude that it is preferable to reduce the overall number of lumbar spine examinations performed, by adherence to accepted guidelines, than to prejudice the detection of significant, albeit rare, abnormalities by restricting the routine series of radiographs.
Discitis↗
The influence of spring and summer New England meteorologic conditions on the respiratory status of patients with chronic lung disease.
To determine whether meteorologic conditions affect the respiratory status of individuals with chronic lung disease, we asked 14 patients who had completed outpatient pulmonary rehabilitation to record the following information daily during the spring and summer of 1991: (1) dyspnea on arising (morning dyspnea); (2) dyspnea throughout the day (daytime dyspnea); (3) peak expiratory flow rate (PEFR); (4) mood; and (5) the presence of lower respiratory tract infection (LRI). The effect of local meteorologic conditions and LRI frequency on morning and daytime dyspnea, PEFR, and mood were analyzed for spring and summer seasons using a linear regression technique that controlled for first-order autocorrelation. The LRI frequency and the preceding day's respiratory status were the most significant predictors of either season's daily variation in respiratory status. During spring, the only meteorologic condition associated with respiratory status was precipitation: rainfall was directly related to increased morning and daytime dyspnea. Meteorologic conditions were not related to changes in PEFR or mood. During summer, the combination of higher temperature, rise in barometric pressure, and increased LRI frequency best predicted increased morning dyspnea, while the combination of higher temperature and rise in barometric pressure best predicted reduced PEFR. Meteorologic conditions were not related to changes in daytime dyspnea or mood. Precipitation in spring and a combination of high temperature and rise in barometric pressure in summer best predicted deterioration in the respiratory status of this group of patients with chronic lung disease.
Aged↗
Hyperplastic polyps: a cell lineage which both synthesizes and secretes trefoil-peptides and has phenotypic similarity with the ulcer-associated cell lineage.
Hyperplastic polyps are common benign lesions of uncertain histogenesis, which occur in the colon in populations at risk for colorectal carcinoma. They contain neutral/MUC1 gene-related mucin which in turn is closely associated with the trefoil-peptide pS2, a major component of the ulcer-associated cell lineage, previously termed pseudopyloric metaplasia. We have examined 17 hyperplastic polyps for expression of the trefoil-peptides pS2 and human spasmolytic polypeptide by in situ hybridization and immunohistochemistry, as well as by using antisera to epidermal growth factor/urogastrone and its receptor and to epitopes of the product of the MUC1 gene to characterize any further similarity between these lesions and the ulcer-associated cell lineage and thus help elucidate the nature of the lesions. Our investigations show both human spasmolytic polypeptide and pS2 messenger RNA within the polyps, whereas only pS2 peptide could be demonstrated immunohistochemically. Epidermal growth factor/urogastrone, its receptor, and antisera to the MUC1 gene also showed widespread staining of these polyps. We suggest that hyperplastic polyps are formed of a lineage that both synthesizes and secretes trefoil-peptides and the MUC1 mucin and that hyperplastic polyps may be related to the phenotypically similar ulceration-associated cell lineage.
Cell Line↗
A discordance between the human muscle NCAM sequence and those seen in other NCAM cDNA clones.
A cDNA clone, designated NC7, has been isolated from human foetal kidney that partially codes for the 140 kDa isoform of human NCAM. This clone contains a 6 bp insert that is not present in the human muscle cDNA clone lambda 4.4. This same sequence has also been found in both a cDNA clone obtained from a human Small Cell Lung Carcinoma (SCLC) line and in human genomic DNA. Furthermore, an equivalent sequence to the 6 bp region identified in the above samples is present in mouse, rat and chicken NCAM. The 6 bp insertion does not lie at a predicted intron/exon boundary as extrapolated by homology studies with the chicken and, therefore, the mechanism by which the sequence is deleted from the human muscle clone lambda 4.4 remains unclear.
Amino Acid Sequence↗
The fate of antibodies bound to the surface of tumor cells in vitro.
The fate of monoclonal antibodies binding to the surface of human tumor cells in vitro was investigated. Seven antibodies, labeled with 125I, were tested on four cell lines, which included a melanoma and carcinomas of the ovary, kidney, and lung. The antibodies were selected only by the criterion that they not be rapidly internalized via coated pits, so that they would be representative of most antibodies reacting with cell surface antigens. After allowing binding during a 2-h incubation, unbound antibody was removed, and the release of intact or degraded antibody in the supernatant was monitored. The data demonstrate that most bound antibody was gradually degraded and released from the cell over a 2-3-day period, probably via internalization, while only a small fraction, less than 20% for most antibodies, appeared to dissociate intact. One exceptional antibody, MW207, dissociated largely intact. The release of intact antibody was virtually complete within 4 h, and radioactivity released after this time was predominantly in degraded form. These results demonstrate that antibody binding to the surface of viable cells must in general be considered irreversible, and hence the concept of affinity is not applicable. Since an Fab fragment of one of the antibodies dissociated rapidly, such irreversible binding appears to require bivalent attachment. Another conclusion of this study is that most antibodies binding to the cell surface are gradually internalized, which we suggest is due to the normal turnover of cell surface constituents via non-clathrin-dependent endocytosis. Several experimental approaches indicated that a large fraction of antibody retained by the cells, for at least 2 days after binding, was present at the cell surface.
Ammonium Chloride↗
Protein kinase C isoforms in multidrug resistant P388/ADR cells: a possible role in daunorubicin transport.
To identify the role of protein kinase C (PKC) isoforms in multidrug resistance in tumor cells, we examined the PKC isoform pattern in the multidrug resistant P388/ADR cell line and studied the effect of down regulation of PKC isoforms on intracellular daunorubicin accumulation and P-glycoprotein expression. Using monoclonal antibodies to PKC alpha, beta and gamma and flow cytometry technique we showed that P388/ADR cells overexpressed PKC alpha and beta as compared to drug sensitive P388 cells. Prolonged treatment of P388/ADR cells with phorbol myristate acetate (PMA), a procedure that is known to down regulate PKC, resulted in the down regulation of total PKC activity and the PKC beta isoform (at the protein level) that was accompanied by the correction of daunorubicin accumulation in P388/ADR cells. The level of expression of P-glycoprotein in PMA treated cells was similar to that of untreated cells. These results suggest that PKC beta regulates the drug efflux function of P-glycoprotein.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Splicing of the VASE exon of neural cell adhesion molecule (NCAM) in human small-cell lung carcinoma (SCLC).
Expression of the neural cell adhesion molecule (NCAM) on small-cell lung carcinoma (SCLC) cell lines and tumour tissue has been investigated. Cell lines were found to express highly sialylated NCAM. Neuraminidase treatment revealed the presence of the 140- and 120-kDa isoforms with differential expression of a 95-kDa protein. Similar data were obtained with SCLC tumour tissues. These results were corroborated by Northern blotting where mRNA of 6.7 and 5.5 kb coding for the 140- and 120-kDa isoforms, respectively, were identified. In a few tumours, a weaker band of 7.4-kb mRNA coding for the 180-kDa NCAM was also identified. This result could not be confirmed biochemically due to shortage of material. Finally, a 5-kb transcript was identified in all SCLC samples examined. The NCAM isoform coded by this mRNA remains unknown. Using the polymerase chain reaction (PCR), we have demonstrated the presence of the VASE mini-exon in some isoforms of SCLC NCAM. The VASE mini-exon sequence in human SCLC differs from the published murine sequence by only one base change. This substitution does not result in altered amino-acid sequence.
Amino Acid Sequence↗
Expression of the neural cell adhesion molecule (NCAM) on the haemopoietic cell line Nalm-16.
The Nalm-16 cell line was originally described as being of the null acute lymphoblastic leukaemia (ALL) phenotype. Using phenotypic and genotypic markers, we have demonstrated the line carries markers associated with cells of the B lineage. In addition, Nalm-16 binds a series of monoclonal antibodies characterized as predominantly recognising neuroectodermal tissues. Amongst these antibodies, UJ13A, 5.1.H11 and ERIC-1 have been shown to recognise the neural cell adhesion molecule (NCAM). Expression of the NCAM molecule is highly complex, with several isoforms of the protein resulting from the differential splicing of the NCAM mRNA transcript. Western-blot analysis of Nalm-16 cell extracts indicates that cells express the heavily polysialylated form of the molecule found on many embryonic tumours. Neuraminidase digestion indicates that the 140 kD isoform is predominantly expressed on Nalm-16, although the 120 kD isoform is present to a lesser degree. These findings have been confirmed using Northern-blot analysis.
Antibodies, Monoclonal↗
X-linked gene MIC5 codes for the L1 adhesion molecule recognized by monoclonal antibody R1.
Monoclonal antibody (MoAb) R1 was generated by use of human-mouse hybrids containing only the long arm of chromosome X as the human component. It recognizes plasma membrane glycoproteins of 145 and 200 kd as well as a soluble protein of 195 kd. In this study, using a biochemical approach, we identified these glycoproteins as the human L1 adhesion molecule.
Animals↗
Identification of region-specific yeast artificial chromosomes using pools of Alu element-mediated polymerase chain reaction probes labeled via linear amplification.
The ability to identify large numbers of yeast artificial chromosomes (YACs) specific to any given genomic region rapidly and efficiently enhances both the construction of clone maps and the isolation of region-specific landmarks (e.g., polymorphic markers). We describe a method of preparing region-specific single-stranded hybridization probes from Alu element-mediated polymerase chain reaction (Alu-PCR) products of somatic cell hybrids for YAC library screening. Pools of up to 50 cloned Alu-PCR products from an irradiation-reduced hybrid containing 22q11.2-q13.1 were labeled to high specific activity by linear amplification using a single vector primer. The resulting single-stranded probes were extensively competed to remove repetitive sequences, while retaining the full complexity of the probe. Extensive coverage of the region by YACs using multiple probe pools was demonstrated as many YACs were detected more than once. In situ analysis using chosen YACs confirmed that the clones were specific for the region. Thus, this pooled probe approach constitutes a rapid method to identify large numbers of YACs relevant to a large chromosomal region.
Base Sequence↗
A comparison of the effects of bid and qid dosing on compliance with inhaled flunisolide.
Noncompliance with inhaled steroids is a well-recognized problem in asthma therapy. We compared compliance with bid and qid regimens of inhaled flunisolide in 16 asthmatic subjects. Patients were instructed to take four inhalations bid for T1 to establish baseline compliance. During T2, half (group A) continued this dosing regimen, while the other half (group B) switched to two inhalations qid. Group A compliance did not significantly change from T1 to T2. The percentage of days with less than eight inhalations (underuse) for group B, however, increased from 20.2 +/- 40.3 in T1 (bid dosing) to 57.1 +/- 49.6 in T2 (qid dosing) (p less than 0.001). Concomitantly, the number of daily inhalations decreased from 7.9 +/- 2.5 to 6.8 +/- 3.1 (p less than 0.01). Reduced compliance with qid dosing was due in large part to an increase in frequency of six inhalations per day, resulting from tid use. Compliance with inhaled flunisolide, therefore, was worse with qid than bid dosing.
Administration, Inhalation↗
An evaluation of severity-modulated compliance with q.i.d. dosing of inhaled beclomethasone.
Although the asthmatic subject's compliance with a regimen of inhaled corticosteroids is often poor, it has been suggested this may improve during periods of increased severity. To test this, we measured daily peak expiratory flow rates (PEFRs), asthma symptoms, and the use of an albuterol inhaler over nine weeks period in ten patients with moderately severe asthma. The effect of changes in these severity indices on compliance with a q.i.d. regimen of inhaled beclomethasone was evaluated. The PEFR was measured in the morning before bronchodilator administration, and symptoms were graded on a scale of 4 to 16, while albuterol and beclomethasone inhalations were electronically recorded. Three measures of compliance with the beclomethasone regimen were used: (1) mean daily compliance ([number of inhalations/number of prescribed inhalations] x 100); (2) underuse, ie, the percentage of days with less than the prescribed number of inhalations; and (3) overuse, ie, the percentage of days with greater than the prescribed number of inhalations. Mean daily compliance was 67 +/- 36 percent, while underuse was observed in 69 percent and overuse in 11 percent of the days. Despite clinical exacerbations in six of the ten patients and considerable variation in the severity indices, no significant relationship was found between the change in asthma severity and compliance with the beclomethasone regimen. These findings do not support the concept of severity-modulated compliance with inhaled corticosteroids.
Administration, Inhalation↗