Semi-quantitative detection of Down's syndrome with PCR.
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Down Syndrome↗
Biomedical subjects
D Miller
Publications and source records attributed to D Miller.
Administration of human recombinant IL-7 to normal and irradiated mice increases the numbers of lymphocytes and some immature cells of the myeloid lineage.
In vitro experiments performed by several investigators have demonstrated that IL-7 is a growth factor for immature B lymphocytes, thymocytes, and mature T lymphocytes. To evaluate the potential therapeutic use for human rIL-7 (rhuIL-7) as a hematopoietin, we have studied the in vivo hematopoietic effects of rhuIL-7 in mice. In these experiments, sublethally irradiated and normal mice were treated with or without rhuIL-7 for up to 26 days. Administration of rhuIL-7 significantly increased the white blood cell count in the peripheral blood and spleen in both normal and irradiated mice. Treatment with rhuIL-7 also accelerated lymphocytic recovery in irradiated mice. Precursor and mature B lymphocytes showed the greatest expansion in response to rhuIL-7 administration, with smaller increases in T lymphocytes being observed. In mice recovering from high dose irradiation, rhuIL-7 treatment resulted in preferential expansion of CD8+ T lymphocytes and more rapid normalization of the CD4/CD8 ratios. Differential analysis of peripheral blood smears demonstrated that rhuIL-7 also increased the numbers of immature granulocytes in both normal and irradiated mice. Moreover, administration of rhuIL-7 to normal, irradiated, cyclophosphamide-pretreated, or 5-fluorouracil-pretreated mice increased the number of acetylcholinesterase-positive megakaryocytes in the spleen, but not the bone marrow. Therefore, although the major in vivo effects of rhuIL-7 were on cells of the lymphocytic lineage, rhuIL-7 also increased the numbers of some immature cells of the myeloid lineage.
Animals↗
Purification and characterization of multiple components of human lymphoblastoid interferon-alpha.
Twenty-two components of human interferon-alpha (IFN-alpha) derived from Sendai virus-induced Namalwa cells were purified by sequential immunoadsorbent affinity chromatography using four monoclonal antibody affinity columns followed by ultrafiltration and reversed-phase high-performance liquid chromatography. The specific activity ranged from 0.2 to 2.6 x 10(8) IU/mg protein on Madin-Darby bovine kidney cells, 0.3 to 4.6 x 10(8) IU/mg protein on human WISH cells, and 10(4) to 7 x 10(5) units/mg protein on mouse L929 cells. The apparent molecular weights of the components ranged from 17,500 to 23,300 using nonreducing sodium dodecyl polyacrylamide-gel electrophoresis and 17,500 to 27,600 using reducing sodium dodecyl polyacrylamide-gel electrophoresis. The amino-terminal amino acid sequences were similar among the components as well as to those reported for the cloned human IFN-alpha genes (Pestka, S. (1986) Methods Enzymol. 119, 3-14). However, four components, f, i, l, and m, have amino-terminal amino acid sequences which appear to be unique when compared to those predicted from the cDNA clones. One component, pre-a, has a potential N-linked glycosylation site on the Asn of residues 2 through 4, Asn-Leu-Ser.
Amino Acid Sequence↗
The 260-kDa transforming growth factor (TGF)-beta binding protein in rat glomeruli is a complex comprised of 170- and 85-kDa TGF-beta binding proteins.
In a previous study (MacKay, K., Robbins, A. R., Bruce, M. D., and Danielpour, D. (1990) J. Biol. Chem. 265, 9351-9356) we showed that rat glomeruli contain transforming growth factor (TGF)-beta 1 binding proteins with apparent molecular masses of 260, 170, and 85 kDa (Gl-260, Gl-170, Gl-85) as determined by electrophoresis under nonreducing conditions. We demonstrate here that Gl-260 is a complex of 170- and 85-kDa TGF-beta binding proteins. Under denaturing conditions the integrity of Gl-260 is maintained through the cross-linking of one monomer of the disulfide-linked TGF-beta 1 homodimer to Gl-85 and of the other monomer to the 100-kDa subunit of Gl-170. In addition, some Gl-260 complexes are maintained by direct cross-linking of Gl-85 to the 100-kDa subunit of Gl-170. One-dimensional peptide maps of Gl-85 and the 100-kDa subunit of Gl-170 indicate that they have distinctly different ligand binding domains. In contrast, peptide maps of Gl-85 and the type II receptor of normal rat kidney fibroblasts are similar. The biological responses of isolated glomeruli to TGF-beta appear to parallel those of cultured glomerular cells which are without detectable Gl-170 and Gl-260 binding proteins.
Affinity Labels↗
Phocine distemper virus outbreak in the Moray Firth common seal population: an estimate of mortality.
Changes in abundance at haul-out sites were followed, and data on the number of deaths collected, to describe the pattern and extent of mortality resulting from the 1988 phocine distemper virus outbreak in the Moray Firth common seal population. Mortality in this population was estimated to be between 10% and 20%, and was low in comparison to some other parts of the North Sea. Most deaths occurred in the 3-month period from 1st August 1988, and there was no sign of a resurgence of the disease during the 1989 breeding season or moult.
Aging↗
Public opinion, the NHS, and the media: changing patterns and perspectives.
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Attitude to Health↗
Quality of life in multiple sclerosis. Comparison with inflammatory bowel disease and rheumatoid arthritis.
Multiple sclerosis (MS) and other chronic illnesses can drastically decrease quality of life (QOL), but there has been little systematic study of QOL in patients with chronic medical diseases. We analyzed QOL in 68 patients with MS, 164 patients with inflammatory bowel disease, and 75 patients with rheumatoid arthritis. The previously validated test instrument was a standardized interview consisting of 41 questions clustered in four subscales: functional and economic scale, social and recreational scale, affect and life in general scale, and medical problems scale. Patients were included in the study if they had a definite medical diagnosis and disease duration of 10 years or longer. In the patients with MS, Kurtzke's Expanded Disability Status Scale correlated strongly only with the medical problems score. Of Kurtzke's Functional System Scales, only the visual Functional System Scores was correlated with total QOL and subscale scores, suggesting that vision is strongly related to QOL. Duration of MS was unrelated to QOL scores. There were significant differences between patients with MS, inflammatory bowel disease, and rheumatoid arthritis on the subscale and total QOL scores. Results suggested that QOL was best in the inflammatory bowel disease group and worst in the MS group. Numerous statistically significant differences on individual questions were evident, suggesting that unique clinical profiles differentially characterize these diseases. Assessments of QOL are a meaningful addition to impairment scales, such as Kurtzke's Expanded Disability Status Scale. Furthermore, QOL scores may meaningfully measure the impact of a chronic medical disease, such as MS, compare the impacts of different diseases, and assess the effects of therapeutic intervention.
Activities of Daily Living↗
Endophthalmitis caused by streptococcal species.
The medical records of 48 patients with culture-positive streptococcal endophthalmitis diagnosed between January 1977 and May 1990 were reviewed. The viridans group streptococci were isolated in 24 (50%) of the 48 cases, enterococci in 13 cases (27.1%), Streptococcus pneumoniae in six cases (12.5%), and beta-hemolytic streptococci in six (12.5%) of 48 cases. The clinical statuses of endophthalmitis cases by etiology were postoperative in 40 patients (83.3%), posttraumatic in six patients (12.5%), and miscellaneous in two patients (4.2%). Overall, 15 (31.2%) patients achieved 20/400 or better visual acuity. The streptococcal isolates demonstrated a 32.6% in vitro resistance to gentamicin sulfate, whereas all isolates were sensitive to vancomycin hydrochloride. The enterococci were often resistant to the cephalosporins, whereas the other streptococcal species were not.
Adolescent↗
Endophthalmitis caused by gram-negative organisms.
The medical records of 52 patients (53 eyes) with culture-proven gram-negative endophthalmitis between January 1982 and December 1990 were reviewed. Pseudomonas aeruginosa (23% [12/53]) and Haemophilus influenzae (19% [10/53]) were the most frequent isolates in this series. Overall, 26 (49%) of 53 treated patients achieved 20/400 or better visual acuity. Fifty-two (98%) of the original 53 gram-negative isolates were sensitive to the aminoglycoside antibiotics. To determine their sensitivity to recently developed antibiotics, 35 of the isolates were again grown on culture media and their sensitivities to ceftazidime, ciprofloxacin, and imipenem were obtained. Only ceftazidime demonstrated in vitro efficacy for all the organisms tested.
Aminoglycosides↗
Rapid detection of gram-negative endotoxin contamination of contact lens saline solutions.
The use of the limulus amoebocyte lysate for the early and rapid detection of gram-negative endotoxin contamination of contact lenses and their solutions could reduce the risk of a keratitis developing that is associated with these devices. Using multiple aliquots from 17 unopened brands of commercially available contact lens saline solutions (15 for soft, two for hard), plus multiple aliquots from these solutions mixed with bacterial endotoxin, we evaluated the ability of two limulus amoebocyte lysate (0.125 endotoxin units/mL) products to detect the presence or absence of gram-negative endotoxin contamination. Sensitivity ranged from 65% (11/17) to 82% (14/17) when the solutions were tested undiluted. When diluted 1:8, the sensitivity increased to 100%. Specificity was 100% for undiluted and diluted specimens. The solutions used for hard contact lenses had the highest false-negative results. The limulus amoebocyte lysate can be used to detect the presence of gram-negative endotoxin in contact lens solutions.
Antimicrobial Cationic Peptides↗
Quantitative MRI of the prostate and uterus in monkeys.
Quantitative MRI has been carried out in the prostate, seminal vesicles, uterus, and ovaries in the pig-tailed monkey, Macaca nemestrina. T2-weighted, fat-suppressed, multislice experiments were performed at 2.35 T. Eight males, 14 ovariectomized females, and 20 intact females were studied. In the prostate, the caudal and cranial lobe were readily distinguished since the latter had a longer T2 value. For all tissues and organs, interanimal variations were large (up to 12-fold variation in volume), but reproducibility was excellent in the prostate and in the ovariectomized monkey uterus with coefficients of variation (CV) of 3 and 5%, respectively. In the intact monkey uterus, cycle-cycle reproducibility was good with CVs of 6-10% in the myometrium and 14-18% in the endometrium. In the follicular phase, endometrial growth (+3.8% day-1, P < 0.001) was accompanied by myometrial shrinkage (-1.6% day-1, P < 0.001), while in the luteal phase, growth was seen in both tissues (+4.3% day-1, P < 0.001 and +1.4% day-1, P < 0.001, respectively). The great value of these MRI techniques in obtaining data in pharmacological efficacy studies of endocrine drugs, and in limiting the number of animals used, is discussed.
Animals↗
Immunological evidence for a conformational difference between recombinant bovine rhodanese and rhodanese purified from bovine liver.
Rhodanese has been utilized as a model enzyme for the study of protein structure-function relationships. The enzyme has recently been cloned and the recombinant enzyme is now available for investigation. However, prior to use in structure-function studies, the recombinant enzyme must be shown to have the same structure and activity as the bovine liver enzyme used in the previous studies. An immunological study of the conformations of these enzyme conformers is described. Three antibodies (two monoclonal and one polyclonal, site-directed antibody) were shown to detect distinct and nonoverlapping epitopes. The epitopes of the monoclonal antirhodanese antibodies (R207 and MAB11) were mapped to the same CNBr digest fragment of the amino terminal domain of rhodanese, and the epitope of the site-directed antibody prepared against the interdomain tether sequence of rhodanese (PAT-T1) was mapped to that region of rhodanese (residues 142-156). The rhodanese conformers were studied by monitoring the accessibility of the epitopes recognized by each antibody in each conformer using an indirect ELISA. None of the antibodies could detect its epitope on the purified liver enzyme. Two of the antibodies (R207 and PAT-T1) could also not detect their epitopes on the recombinant enzyme. However, MAB11 did detect a conformational difference between the natural and recombinant rhodanese conformers, indicating the conformational difference is localized in the first 73 amino acids of rhodanese. This difference presumably reflects the difference in the histories of the two enzymes and may be due to differences in enzyme folding, differences in the purification procedures, and differences in storage conditions--all of which could influence the final conformation of the enzyme.
Amino Acid Sequence↗
Expectant management of choriocarcinoma limited to placenta.
Choriocarcinoma limited to placenta was discovered "incidentally" following a seemingly uneventful term pregnancy. The newborn had unsuspected severe anemia and thrombocytopenia, due to fetomaternal hemorrhage. His recovery was good following transfusions. The mother was asymptomatic and her extensive workup for metastatic choriocarcinoma was negative. She was subsequently managed expectantly and monitored with serial serum beta-hCG, demonstrating near-logarithmic decline to non-pregnant levels within 5 weeks of delivery.
Adult↗
A high-resolution meiotic mapping panel for the pericentromeric region of chromosome 10.
Familial multiple endocrine neoplasia type 2A (MEN 2A) is a dominantly inherited cancer syndrome characterized by tumors in tissues derived from the neural crest. The disease manifests as medullary carcinoma of the thyroid, pheochromocytoma, and hyperparathyroidism. The MEN2A locus has been mapped near the centromere of chromosome 10 by linkage analysis. Statistical analyses have not resolved the location of MEN2A among several close markers. We have used our family material to refine the positions of 36 identified and confirmed crossovers among the markers most closely linked to MEN2A. This high-resolution meiotic mapping panel will help order loci in this pericentromeric region and narrow the region in which MEN2A maps.
Centromere↗
Comparison of immunocytology to tissue culture for diagnosis of presumed herpesvirus dendritic epithelial keratitis.
OBJECTIVE: The objective of this study is to prospectively compare the sensitivity and specificity of immunodetection of herpes simplex virus (HSV) in impression cytology specimens obtained directly from presumed herpesvirus dendritic epithelial keratitis with virus isolation by tissue culture of cells scraped from the same lesion. METHODS: Corneal impression cytology and tissue culture were performed on 29 consecutive patients presenting with presumed herpesvirus dendritic epithelial keratitis during a 6-month period. Impression cytology of dendritic epithelial keratitis lesions with Millipore Biopore membranes were evaluated for the presence of antigens specific to HSV type I (HSV-1), HSV-2, and varicella-zoster virus (VZV) using monoclonal antibodies specific to these herpesviruses and immunofluorescent staining techniques. RESULTS: Tissue culture was positive for HSV-1 in 52% (13 of 25) of dendritic epithelial keratitis patients without skin lesions, and was negative for VZV in 4 patients with dendritic epithelial keratitis and skin lesions in the distribution of the first division of the trigeminal nerve. The remaining 12 tissue cultures showed no cytopathic effect. Compared with tissue culture, impression cytology was 100% sensitive (13 of 13) and 92% specific (11 of 12) for the diagnosis of HSV-1 dendritic epithelial keratitis (Kappa coefficient of agreement 0.92). Although our sample size for VZV dendritic epithelial keratitis was small, the impression cytology findings correlated with our clinical diagnosis more often than tissue culture (2 of 4 versus 0 of 4). CONCLUSION: Impression cytology allows simultaneous debridement of dendritic epithelial keratitis and, when combined with immunocytologic staining procedures, provides a simpler, more rapid, and less expensive alternative to tissue culture for the diagnosis of dendritic epithelial keratitis caused by HSV or VZV.
Antibodies, Viral↗
Production of extracellular matrix by glomerular epithelial cells is regulated by transforming growth factor-beta 1.
Transforming growth factor-beta (TGF-beta) has widespread effects on extracellular matrix production by many cultured cell lines and appears to play a role in the pathological accumulation of extracellular matrix that accompanies inflammatory and fibrotic diseases such as glomerulonephritis. Earlier experiments have shown that mesangial cells respond to TGF-beta 1 with a marked increase in the production of two chondroitin/dermatan sulfate proteoglycans, decorin and biglycan, but their production of other matrix components elevated in glomerulonephritis is not substantially affected by TGF-beta 1. Since the glomerular epithelial cells are also thought to contribute to matrix production in the glomerulus, we examined the ability of these cells to produce some of the nonproteoglycan matrix components in response to TGF-beta 1. Exposure of glomerular epithelial cells to TGF-beta 1 increased the production of fibronectin and type IV collagen, in addition to biglycan. Enhancement of the cell layer accumulation of laminin was also observed. These results show that TGF-beta 1 has a differential effect on extracellular matrix production by epithelial and mesangial cells from glomeruli. TGF-beta 1 released in the glomerulus secondary to injury could thus affect both cell types and lead to increased intraglomerular production of proteoglycans, whereas the increased fibronectin, type IV collagen, and laminin may primarily originate from the epithelial cells.
Animals↗
Genotypes and sequence variants of human papillomavirus DNAs from human immunodeficiency virus type 1-infected women with cervical intraepithelial neoplasia.
Human papillomavirus (HPV) DNA was found in cervicovaginal lavage fluids from 9 of 11 human immunodeficiency virus type 1 (HIV-1)-seropositive female prostitutes with cervical intraepithelial neoplasia (CIN) in Kinshasa, Zaire. Since 7 yielded complex nucleic acid hybridization results consistent with mixed HPV infections, limited sequencing of HPV DNA was used to identify the HPVs present. Three of HPV 16 and 1 each of HPV 18, 31, 33, and 56 and ME180-HPV were identified by sequencing in 8 samples. Each of these genotypes has been found in specimens from HIV-1-seronegative women with CIN. Some DNAs had nucleic acid and amino acid sequence variations relative to the reference HPVs, but the variants were closely related to variants that have been found in HIV-1-seronegative women. Variant amino acids were found predominantly at three positions in one 40-amino-acid segment of the L1 open reading frame sequenced. The predominant HPV 16 variant observed has been found rarely in other countries.
Amino Acid Sequence↗
Characterization and cellular distribution of human spermatozoal heat shock proteins.
Heat shock proteins (hsp) are ubiquitous components of all living systems. They are up-regulated in response to adverse changes in the cellular environment and at least one highly conserved group (hsp 70) is associated with the development of tolerance to various physico-chemical stress inducers. Spermatozoa have highly condensed chromatin and unlike somatic cells, are consequently unable to mount a stress response. However, using a combination of gel electrophoresis and immunoblotting with hsp-specific monoclonal antibodies, we report that proteins of M(r) 95 kDa and 70-75 kDa corresponding to hsp 90, and multiple forms of hsp 70 are present in human spermatozoa. Immunohistochemistry localized hsp 90 to the neck and tail of unfixed, acrosome-intact spermatozoa. In contrast, an equatorial ring surrounding the nucleus was observed in unfixed spermatozoa, acrosome-reacted with the calcium ionophore A23187. The ring was stained in cells fixed and permeabilized with ethanol, regardless of acrosomal status. Hsp 70 was an abundant surface antigen and as this protein was also abundant in seminal plasma, we believe that it may have been directly adsorbed onto the cell surface. More specific midpiece, equatorial and nuclear staining was also observed. Possible functions for spermatozoal heat shock proteins are discussed.
Antibodies, Monoclonal↗