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1.  Modeling the Matrix-Cilium-Golgi continuum in hyaline chondrocytes by electron tomography 
Cilia  2012;1(Suppl 1):P39.
PMCID: PMC3555806
3.  Differential Cytokine Responses in Human and Mouse Lymphatic Endothelial Cells to Cytokines in Vitro 
Lymphatic Research and Biology  2010;8(3):155-164.
Inflammatory cytokines dysregulate microvascular function, yet how cytokines affect lymphatic endothelial cells (LEC) are unclear.
Methods and Results
We examined effects of TNF-α, IL-1β, and IFN-γ on LEC proliferation, endothelial cell adhesion molecule (ECAM) expression, capillary formation, and barrier changes in murine (SV-LEC) and human LECs (HMEC-1a).
All cytokines induced ICAM-1, VCAM-1, MAdCAM-1, and E-selectin in SV-LECs; TNF-α, IL-1β and IFN-γ induced ECAMs (but not MAdCAM-1) in HMEC-1a. IL-1β increased, while IFN-γ and TNF-α reduced SV-LEC proliferation. While TNF-α induced, IFN-γ decreased, and IL-1β did not show any effect on HMEC-1a proliferation. TNF-α, IL-1β, and IFN-γ each reduced capillary formation in SV-LEC and in HMEC-1a. TNF-α and IL-1β reduced barrier in SV-LEC and HMEC-1a; IFN-γ did not affect SV-LEC barrier, but enhanced HMEC-1a barrier. Inflammatory cytokines alter LEC growth, activation and barrier function in vitro and may disturb lymphatic clearance increasing tissue edema in vivo.
Therapies that maintain or restore lymphatic function (including cytokines blockade), may represent important strategies for limiting inflammation.
PMCID: PMC3357074  PMID: 20863268
4.  Ichthyosis in Sjögren–Larsson syndrome reflects defective barrier function due to abnormal lamellar body structure and secretion 
Archives of Dermatological Research  2010;302(6):443-451.
Sjögren–Larsson syndrome is a genetic disease characterized by ichthyosis, mental retardation, spasticity and mutations in the ALDH3A2 gene coding for fatty aldehyde dehydrogenase, an enzyme necessary for oxidation of fatty aldehydes and fatty alcohols. We investigated the cutaneous abnormalities in 9 patients with Sjögren–Larsson syndrome to better understand how the enzymatic deficiency results in epidermal dysfunction. Histochemical staining for aldehyde oxidizing activity was profoundly reduced in the epidermis. Colloidal lanthanum perfusion studies showed abnormal movement of tracer into the extracellular spaces of the stratum corneum consistent with a leaky water barrier. The barrier defect could be attributed to the presence of abnormal lamellar bodies, many with disrupted limiting membranes or lacking lamellar contents. Entombed lamellar bodies were present in the cytoplasm of corneocytes suggesting blockade of lamellar body secretion. At the stratum granulosum–stratum corneum interface, non-lamellar material displaced or replaced secreted lamellar membranes, and in the stratum corneum, the number of lamellar bilayers declined and lamellar membrane organization was disrupted by foci of lamellar/non-lamellar phase separation. These studies demonstrate the presence of a permeability barrier abnormality in Sjögren–Larsson syndrome, which localizes to the stratum corneum interstices and can be attributed to abnormalities in lamellar body formation and secretion.
PMCID: PMC2892059  PMID: 20049467
Ichthyosis; Fatty alcohol; Fatty aldehyde; Aldehyde dehydrogenase; Stratum corneum; Genetic disease
5.  The Phase-Variable Allele of the Pilus Glycosylation Gene pglA Is Not Strongly Associated with Strains of Neisseria gonorrhoeae Isolated from Patients with Disseminated Gonococcal Infection▿ †  
Infection and Immunity  2007;75(6):3202-3204.
The Neisseria gonorrhoeae pglA gene has two alleles, one of which is phase variable. A previous study reported that all disseminated gonococcal infection (DGI) isolates contained the phase-variable allele and proposed a causal link. In the present study of 81 strains no absolute correlation between DGI and the phase-variable pglA allele was observed.
PMCID: PMC1932893  PMID: 17296763
6.  Assessing the accuracy of an automated coding system in emergency medicine. 
Accuracy and speed are imperative when it comes to coding medical records. Completely automated approaches to coding are faster than human coders, but are they as accurate? To measure accuracy, a "gold standard" is required; however, establishing a standard for medical records coding is problematic given the inherent ambiguity in some of the coding rules and guidelines. This paper presents statistics regarding the variability amongst experienced coders and compares this variability with an automated system, LifeCode. The authors conclude that LifeCode is as accurate as the human coders used in this study and offers the potential for increased coding consistency and productivity.
PMCID: PMC2244078  PMID: 11079953
9.  Detection of human immunodeficiency virus type 1 RNA in plasma samples from high-risk pediatric patients by using the self-sustained sequence replication reaction. 
Journal of Clinical Microbiology  1992;30(2):281-286.
There is an urgent need for rapid and sensitive methods to assess human immunodeficiency virus (HIV) infection in infants and children. We evaluated an approach by using the self-sustained sequence replication reaction (3SR) to amplify HIV type 1 (HIV-1) RNA directly. The amplified RNA product was then detected by bead-based sandwich oligonucleotide capture hybridization and rare earth metal chelate time-resolved fluorescence. The sensitivity of this technology was determined to be less than 12 HIV-1 RNA copies with an amplification level of 10(10)-fold with purified HIV-1 RNA. Plasma samples from 19 high-risk pediatric patients younger than 5 years of age were examined, and results were compared with viral culture of patient plasma. Results from plasma culture and 3SR amplification agreed for 14 of these patients and disagreed for 5. Of the five samples which did not agree, four were positive by 3SR and negative by culture and one was positive by culture and negative by 3SR but became positive by 3SR at a subsequent testing. We conclude that 3SR amplification coupled with time-resolved fluorescence is a promising technology for investigating the relationship between the presence of HIV-1 RNA in plasma and progression of disease in HIV-infected pediatric patients. This technology should be important in the assessment of HIV-1 infection, in evaluating drug therapies, and in understanding the pathogenesis and transmission of the virus.
PMCID: PMC265046  PMID: 1537893
10.  A highly divergent simian immunodeficiency virus (SIVstm) recovered from stored stump-tailed macaque tissues. 
Journal of Virology  1991;65(12):7061-7065.
We report here the results of molecular analysis of a simian immunodeficiency virus (designated SIVstm) which was isolated from a rhesus monkey inoculated with stored lymph node tissue of an Asian stump-tailed macaque. The latter monkey had died in 1977 during an epidemic of acquired immunodeficiency and lymphoma at the California Regional Primate Research Center (L. J. Lowenstine, N. W. Lerche, P. A. Marx, M. B. Gardner, and N. C. Pedersen, p. 174-176, in M. Girard and L. Valette, ed., Retroviruses of Human AIDS and Related Animal Viruses, 1988). Nucleotide sequence analysis of the gag and env regions indicates that SIVstm is an ancient member of the SIV/human immunodeficiency virus type 2 group; it is quite divergent from known SIVs isolated from African sooty mangabeys as well as from Asian macaques. Furthermore, of all SIV strains described to date, SIVstm is the most closely related to human immunodeficiency virus type 2.
PMCID: PMC250828  PMID: 1942258
11.  Audit of a new appointments system in a hospital outpatient clinic. 
BMJ : British Medical Journal  1991;302(6769):148-149.
OBJECTIVE--To assess the effect of a new appointments system on patients' waiting time. DESIGN--Replacement of an existing system of regularly spaced appointments at 10 minute intervals with one in which the doctor arranged appointments according to his perception of individual patients' requirements, from December 1988 to June 1989. SETTING--One general medical outpatient clinic. PATIENTS--All (181) [corrected] patients under regular review with fixed appointment times during six months; those arriving for blood testing before the start of the clinic and those requiring ambulance transport were excluded. MAIN OUTCOME MEASURES--Mean waiting time (mean of difference between appointment time and start of consultation), maximum waiting time, number of patients, and total duration of clinics. RESULTS--Mean waiting time was reduced by about 30 minutes (from 39.6 to 9.5 mins) over the six months. There was no change in the number of patients attending (mean 15.8 v 15.4) or the duration of the clinics (mean 187 v 160 mins). CONCLUSION--Efficiency was improved by simple adjustment to the appointments system used in the hospital outpatient clinic.
PMCID: PMC1668800  PMID: 2009103
12.  Inactivated simian immunodeficiency virus vaccine failed to protect rhesus macaques from intravenous or genital mucosal infection but delayed disease in intravenously exposed animals. 
Journal of Virology  1990;64(5):2290-2297.
Eight rhesus macaques were immunized four times over a period of 8 months with a psoralen-UV-light-inactivated whole simian immunodeficiency virus vaccine adjuvanted with threonyl muramyl dipeptide. Eight unvaccinated control animals received adjuvant alone. Only the vaccinated animals made antibodies before challenge exposure to the viral core and envelope as determined by Western blotting (immunoblotting) and virus-neutralizing antibodies. Ten days after the final immunization, one-half of the vaccinated and nonvaccinated monkeys were challenged exposed intravenously (i.v.) and one-half were challenge exposed via the genital mucosa with virulent simian immunodeficiency virus. All of the nonvaccinated control monkeys became persistently infected. In spite of preexisting neutralizing antibodies and an anamnestic antibody response, all of the immunized monkeys also became persistently infected. However, there was evidence that the clinical course in immunized i.v. infected animals was delayed. All four mock-vaccinated i.v. challenge-exposed animals died with disease from 3 to 9 months postchallenge. In contrast, only one of four vaccinated i.v. challenge-exposed monkeys had died by 11 months postchallenge.
PMCID: PMC249390  PMID: 2157886
13.  Analysis of the Escherichia coli gene encoding L-asparaginase II, ansB, and its regulation by cyclic AMP receptor and FNR proteins. 
Journal of Bacteriology  1990;172(3):1491-1498.
Escherichia coli contains two L-asparaginase isozymes: L-asparaginase I, a low-affinity enzyme located in the cytoplasm, and L-asparaginase II, a high-affinity secreted enzyme. A molecular genetic analysis of the gene (ansA) encoding the former enzyme has previously been reported. We now present a molecular study of the gene, ansB, encoding L-asparaginase II. This gene was isolated by using oligonucleotide probes, whose sequences were based on the previously determined amino acid sequence. The nucleotide sequence of ansB, including 5'- and 3'-untranslated regions, was determined. The amino acid sequence of L-asparaginase II, deduced from this nucleotide sequence, contains differences at 11 positions when compared with the previously determined amino acid sequence. The deduced amino acid sequence also reveals a typical secretory signal peptide of 22 residues. A single region of sequence similarity is observed when ansA and ansB are compared. The transcriptional start site in ansB was determined, allowing the identification of the promoter region. The regulation of ansB was studied by using ansB'-'lacZ fusions, together with a deletion analysis of the 5' region upstream of the promoter. Regulation by cyclic AMP receptor protein and anaerobiosis (FNR protein) was confirmed, and the presence of nucleotide sequence motifs, with homology to cyclic AMP receptor protein and FNR protein-binding sites, investigated.
PMCID: PMC208625  PMID: 2407723
14.  Genital mucosal transmission of simian immunodeficiency virus: animal model for heterosexual transmission of human immunodeficiency virus. 
Journal of Virology  1989;63(10):4277-4284.
An animal model for the heterosexual transmission of human immunodeficiency virus (HIV) was developed by the application of simian immunodeficiency virus (SIV) onto the genital mucosas of both mature and immature, male and female rhesus macaques. Virus preparations were infused into the vaginal vaults or the urethras (males) of the animals through a soft plastic pediatric nasogastric feeding tube. The macaques that were infected by this route (six males and nine females) developed SIV-specific antibodies, and SIV was isolated from peripheral mononuclear cells of all seropositive animals. One male and one female infected by this route developed severe acquired immunodeficiency syndrome-like disease with retroviral giant-cell pneumonia. As few as two inoculations of cell-free SIV containing 50 50% tissue culture infective doses induced persistent viremia. Cell-free virus preparations were capable of producing infection by the genital route. Much higher doses of virus were required to transmit SIV by this route than are required for transmission by intravenous inoculation. Thus, it appears that the mucous membranes of the genital tract act as a barrier to SIV infection. Spermatozoa and seminal plasma were not required for the genital transmission of SIV. Rarely, SIV was recovered from mononuclear cells in semen and vaginal secretions. The SIV-rhesus macaque model is suitable for assessing the role of cofactors in heterosexual transmission of HIV and will be useful for testing the effectiveness of spermicides, pharmacologic agents, and vaccines in preventing the heterosexual transmission of HIV.
PMCID: PMC251042  PMID: 2778875
15.  Antigen detection for human immunodeficiency virus. 
Clinical Microbiology Reviews  1989;2(3):241-249.
The recent development of enzyme immunoassay procedures for the direct determination of human immunodeficiency virus (HIV) antigens has been of significant benefit in both clinical and research applications. The historical development of HIV antigen assays as well as their current and future applications for use in the clinical microbiology laboratory are reviewed. A detailed description of selected commercially available assays is presented, and a comparison is made of various parameters, including sensitivity, specificity, and cost. The use of the HIV antigen assay as an alternative to the reverse transcriptase assay in virus culture applications is also discussed. In addition, the diagnostic and prognostic utility of the HIV antigen assay is considered for various patient groups, including neonatal, high-risk asymptomatic, seronegative, and seropositive patient populations. The use of the HIV antigen assay as an adjunct to anti-HIV antibody testing, as well as its utility in assessing the therapeutic efficacy of antiviral drug therapy, is discussed. The biology of HIV antigen expression and modulation of anti-HIV antibody titers during infection are also discussed in terms of two possible models.
PMCID: PMC358118  PMID: 2670189
16.  Blue rubber bleb naevus disease: an uncommon cause of gastrointestinal tract bleeding. 
Gut  1988;29(10):1408-1412.
A 23 year old woman presented with facial pain, a right parotid tumour and iron deficiency anaemia. She had several cutaneous venous swellings and tumours with a similar appearance were found in the large bowel. Histological examination of the parotid tumour and angiography of the skin and gut lesions confirmed that they were venous in origin. The aetiology, classification, and complications of disorders of the venous system and the importance of using a tourniquet to examine the peripheral veins is discussed.
PMCID: PMC1434013  PMID: 3264257
17.  Immunological abnormalities 17 years after accidental exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. 
Eighteen workers were reviewed 17 years after accidental exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin). Clinical assessment showed that they were in good health. A study of several biochemical and immunological parameters in these subjects and in 15 carefully matched controls showed no difference in serum concentrations of hepatic enzymes between exposed workers and controls. Although mean serum concentrations of cholesterol and triglyceride were higher in exposed subjects than in controls, the results did not reach statistical significance. Antinuclear antibodies and immune complexes were detected significantly more frequently in the peripheral blood of workers exposed to dioxin. There was no significant difference between exposed workers and controls in the number of T lymphocytes, B lymphocytes, and helper and suppressor T cell counts in peripheral blood, but the number of natural killer cells identified by the monoclonal antibody Leu-7 was significantly higher in workers exposed to dioxin.
PMCID: PMC1009680  PMID: 3264183
18.  Unique p24 epitope marker to identify multiple human immunodeficiency virus variants in blood from the same individuals. 
Journal of Clinical Microbiology  1987;25(8):1411-1415.
The human immunodeficiency virus (HIV) was isolated from the blood of 192 of 410 seropositive individuals. Original isolations were made in peripheral blood mononuclear cell (PBMC) cultures, and only one-fifth of the HIV isolates could be adapted to replicate in continuous T-cell lines. Of the 192 HIV isolates, 42 had the characteristic p24 antigen marker of the acquired immunodeficiency syndrome-associated retrovirus type 2 strain of HIV (HIV ARV-2) and 150 resembled the human T-cell lymphotropic virus type III strain of HIV (HIV HTLV-III). Significantly, primary PBMC cultures from two patients yielded multiple variants. When these variants were exposed to continuous T-cell lines, only one of them continued to replicate. The remaining variants were lost and could not be reisolated following passage back into PBMC cultures. We conclude the following from these studies: PBMC cultures are more efficient at isolating HIV than continuous T-cell lines are; some patients harbor more than one genetic variant of HIV in the blood at the same time; and continuous T-cell lines are likely to yield only a portion of the HIV variants originally present in the blood.
PMCID: PMC269236  PMID: 2442190
20.  Practice of preoperative assessment by anaesthetists. 
The practice of preoperative assessment in 24 departments of anaesthesia in Great Britain and Ireland was surveyed. Most departments had no rigid policies governing assessment, and many served several hospitals. There was little evidence that admission procedures of patients scheduled for surgery or the organisation of operating lists took account of the problems encountered by anaesthetists undertaking preoperative assessment. From the participating departments 415 anaesthetists completed a questionnaire of their individual practice. Most (57%) visited at least 80% of their patients preoperatively, but 22% saw less than 50% of patients. The detection of potential anaesthetic problems and the establishment of rapport with patients were highly rated reasons for conducting such visits. Failure to visit was often related to organisational defects within the hospital service, and anaesthetists saw little prospect of improving these defects. The demands created by the needs of preoperative assessment on the one hand, and the need for a rapid turnover of surgical patients and financial stringency on the other, conflict, and this conflict is not easily reconciled.
PMCID: PMC1416463  PMID: 3926208
21.  Severe hyperkalaemia and ketoacidosis during routine treatment with an insulin pump. 
During a feasibility study of the use of insulin pumps to treat diabetes ketoacidosis occurred at a rate of 0.14 episodes/patient/year in the first year but was lower in subsequent years. A case of cardiac arrest secondary to hyperkalaemia during ketoacidosis occurred in a patient treated with a pump. The mean (SD) serum potassium concentration on presentation to hospital with ketoacidosis was significantly higher in patients treated with a pump (5.7 (1.1) mmol(mEq)/l) than those treated with conventional injections of insulin (4.9(0.9) mmol/l; p less than 0.01). The high rate of ketoacidosis and raised serum potassium concentrations during treatment with the pump creates doubt about the use of this treatment as an alternative regimen for large numbers of patients in a busy diabetic clinic.
PMCID: PMC1416428  PMID: 3926198
25.  Analysis of an inversion within the human beta globin gene cluster. 
Nucleic Acids Research  1985;13(8):2897-2906.
We have cloned and sequenced the DNA from two regions of the defective beta-globin gene cluster from a patient with Indian A gamma delta beta thalassaemia, and confirmed the complex and unusual pattern of rearrangement involving two separate deletions (0.8 kb and 7.5 kb) the inversion of the 15.5 kb segment separating them, as previously proposed from gene mapping studies [1]. All four breakpoints occur within the transcribed region of the globin genes and at one junction are found six nucleotides of unknown origin. This unique rearrangement results in enhanced expression of the upstream fetal gene, and is therefore is pertinent to the localisation of any putative control region involved in the coordinate expression of fetal and adult genes.
PMCID: PMC341202  PMID: 4000967

Results 1-25 (28)