Hyperpnoea and emesis in a diabetic man.
Explore the source record for details and available documents.
Adult↗
Biomedical subjects
D Smith
Publications and source records attributed to D Smith.
Baroregulation of vasopressin release in adipsic diabetes insipidus.
Adipsic diabetes insipidus (ADI) occurs in association with a heterogeneous group of conditions. We report vasopressin (AVP) responses to hypotension in nine patients with ADI and nine controls. Hypertonic saline infusion produced absent thirst (1.7 +/- 1.7 to 1.5 +/- 1.7 cm, P = 0.99) and AVP responses (0.3 +/- 0.1 to 0.4 +/- 0.1 pmol/liter, P = 0.99) in the ADI group, who also drank less than the control group (258 +/- 200 ml vs. 1544 +/- 306 ml, P < 0.001). Intravenous infusion of trimetaphan camsylate produced a fall in mean arterial pressure of 31.6% +/- 8.9% in patients and 29.4% +/- 6.1% in controls. Plasma AVP concentrations rose from 1.4 +/- 0.8 to 340.3 +/- 497.4 pmol/liter (P < 0.001) in the control group. In three patients with craniopharyngioma, there was no rise in plasma AVP concentrations (0.3 +/- 0.1 to 0.3 +/- 0.1 pmol/liter, P = 0.96), but plasma AVP rose significantly in response to hypotension in the other six patients (0.4 +/- 0.2 to 204.5 +/- 223.2 pmol/liter, P < 0.001). We concluded that the AVP responses to hypotension in ADI are heterogeneous and reflect the site of the lesion causing the diabetes insipidus.
Adolescent↗
Fully automated radioligand binding filtration assay for membrane-bound receptors.
Here we describe a fully automated, hands-free radioligand filtration binding assay for dopamine D3 receptors. Three separate instruments were linked in tandem to perform the following operations: The Genmate and Genesis were linked to perform liquid handling, incubation, and the scheduling operations, while an automated harvester was used to perform rapid filtration. To minimize carryover of compounds, disposable tips were used for diluting and dispensing the compounds. A custom-designed tip holder was used to handle loading and pipetting by the Genmate 96-well pipettor. The assay for 84 compounds with six concentrations that spans six logs can be completed within 4 h. The reproducibility of the individual data point (cv < 10% between duplicates) and Ki (cv < 20%) is superior to that determined by manual procedures. Ki values of various dopamine ligands determined by the hands-free procedure are similar to published values. This technology reduces hands-on time (at least 70%), minimizes exposure to radioligands (up to 95%), and improves the reproducibility of results. The technology is applicablefor high-throughput screening and rapid determination of structure-activity relationship of compounds for many other membrane-bound receptors.
Animals↗
A Monte-Carlo estimation of effect size distortion due to significance testing.
A Monte-Carlo study was done with true effect sizes in deviation units ranging from 0 to 2 and a variety of sample sizes. The purpose was to assess the amount of bias created by considering only effect sizes that passed a statistical cut-off criterion of alpha = .05. The deviation values obtained at the .05 level jointly determined by the set effect sizes and sample sizes are presented. This table is useful when summarizing sets of studies to judge whether published results reflect an accurate appraisal of an underlying effect or a distorted estimate expected because significant studies are published and nonsignificant results are not. The table shows that the magnitudes of error are substantial with small sample sizes and inherently small effect sizes. Thus, reviews based on published literature could be misleading and especially so if true effect sizes were close to zero. A researcher should be particularly cautious of small sample sizes showing large effect sizes when larger samples indicate diminishing smaller effects.
Humans↗
Estimation of pedigree errors in the UK dairy population using microsatellite markers and the impact on selection.
The proportion of cows in the UK dairy herd whose sires were misidentified was estimated using DNA markers. Genetic marker genotypes were determined on 568 cows (from 168 milk samples and 400 hair samples) and 96 putative sires (from semen samples). The estimated pedigree error rate from the hair samples was 8.8%, and from the milk samples, 13.1%, giving an overall estimate of the error rate of 10%. This level of pedigree errors will have a relatively large impact on the efficiency of progeny testing and the accuracy of cow predicted breeding values. We predict a loss of response to selection of approximately 2 to 3% given this error rate.
Animals↗
The establishment of a hospital clinical ethics committee.
In the Republic of Ireland the establishment of Clinical Ethics Committees (CECs) are relatively new. The need for such committees has occurred due to the fact that the consideration of ethical issues in healthcare has become an important and frequent part of discussions by individuals and institutions. A number of factors have contributed to this growth of ethical considerations. The Bon Secours Health System (BSHS) decided to establish a CEC and appointed a co-ordinating team to draw up terms of reference for such a committee. During this process the co-ordinating team drew on the experience of other countries and of its own staff. Potential criticisms of how such a committee would function were examined. A representative membership of the CEC was also arrived at. The following functions were identified for a CEC: the need to provide a mechanism for the identification, discussion and resolution of medical ethical issues; the need to identify medical ethical issues which may create challenges to the health system and to monitor the responses of the health system to these areas, and the provision of education. Policy development was seen as a primary function of the CEC. Ethical case review also emerged as one area of potential involvement by the CEC. During the staff consultation a large number of ethical issues emerged which the staff requested the CEC to address. A methodology necessary for coping with the differences within the BSHS was developed by the CEC. It is evident that CECs are here to stay and how they develop and function will have an impact on the quality of healthcare.
Ethics Committees, Clinical↗
The anatomy of the human hypoglycaemia sensor.
Explore the source record for details and available documents.
Animals↗
Warm autoimmune hemolytic anemia with mimicking anti-c and -E specificities.
An 18-month-old male was admitted to a hospital with a hemoglobin of 4.1 g/dL and a reticulocyte count of 53 percent. There was no history of prior transfusion. Serologic evaluation revealed the presence of both a positive direct antiglobulin test (DAT) and an indirect antiglobulin test (IAT). The patient's red blood cells (RBCs) typed as group A, C-D-E-c+e+ (cde/cde). Evaluation of the IAT revealed the presence of anti-c and anti-E. All other major antibodies were ruled out. Upon adsorption of the patient's serum with ficin-treated Cde/Cde RBCs, both antibody specificities were adsorbed; however, the antibodies were not adsorbed with native (untreated) Cde/Cde RBCs. Furthermore, the autoantibody was not adsorbed by Rhnull cells, thereby suggesting Rh specificity. The serum was incompatible with cde/cde RBCs and compatible with Cde/Cde RBCs. The patient was successfully transfused with Cde/Cde RBCs followed by resolution of his anemia, as evidenced by an increased and stable hemoglobin. It was concluded that the autoantibody had mimicking anti-c and -E specificities. This is a report of an unusual case of autoimmune hemolytic anemia because the Rh autoantibody appeared to have dual mimicking specificities, and the patient's RBCs were antigen negative for one of the antibody specificities, i.e., anti-E.
Journal Article↗
Mapping the actin filament with myosin.
Structural studies have shown that the heads of the myosin motor molecule bind preferentially to "target zones" of favorably oriented sites on the helices of the actin filament. We present direct evidence for target zones from the interactions of a single myosin head with an actin filament held between two optically trapped beads. With compliant traps, thermal motions of the filament allow the fixed myosin-S1 to interact with at least two zones, observed as a bi-modal distribution of filament displacements due to myosin binding, whose spacing is near the 36-nm helix repeat distance. The number of binding events and the "apparent working stroke" (mean displacement with myosin bound), vary periodically as the filament is moved past the fixed myosin by displacing the traps; observed periods are close to 36 nm and the apparent stroke varies from 0-10 nm. We also observe a strong modulation at the 5.5-nm actin monomer repeat confirming that myosin interacts with a single strand and that the actin is not free to rotate. Each interaction can be assigned to an actin monomer and each active zone on the helix is made up of three actin monomers.
Actins↗
Whole cell ELISA for detection of tumor antigen expression in tumor samples.
We have developed a whole cell enzyme-linked immunosorbent assay (ELISA) that detects the melanoma antigens gp100 and S-100 in tumor samples from patients with metastatic melanoma and the antigen CA 125 in tumor samples from patients with ovarian carcinoma. The assay is relatively simple to perform and interpret without specialized expertise in pathology. It is both sensitive and selective and is amenable to being automated. The results correlate very well with those obtained by flow cytometry and are in good accord with published values obtained by immunohistochemistry. We believe that this assay should be very helpful for characterizing autologous cell cancer vaccines and may also represent a useful alternative to immunohistochemistry for cancer diagnosis. It should be adaptable to other types of cancer where tumor antigens have been identified and good antibodies are available.
Antigens, Neoplasm↗
Generation of recombinant human C3dg tetramers for the analysis of CD21 binding and function.
CD21 (complement receptor 2, CR2) binds the terminal proteolytic fragments of the third component of complement (C3) that have been covalently attached to immune complexes or other targets during the activation of complement. We used the technique of in vivo biotinylation to create a recombinant multivalent ligand for CD21. A sequence coding for a biotinylation signal peptide was added to the 3' end of the human C3dg cDNA. The modified C3dg was expressed in Escherichia coli and biotinylated intracellularly by the bacterial biotin holoenzyme synthetase (BirA) enzyme. Monomeric C3dg was unable to bind to CD21 as determined by flow cytometry, while biotinylated recombinant C3dg (rC3dg) complexed with fluorochrome-conjugated streptavidin bound tightly. Binding was observed using CD21 positive B cells but not seen on pre-B cells that do not express this complement receptor. Two assays were used to assess the functional capacity of the recombinant C3dg. First, multimeric C3dg caused the phosphorylation of the mitogen-activated kinase, p38, in mature B lymphoma cells. Second, C3dg greatly enhanced the activation of primary B cells in combination with a sub-stimulatory concentration of anti-IgM monoclonal antibody. These results illustrate the utility of the technique of in vivo biotinylation to generate ligands for cell surface receptors that require multimerization for high avidity binding and function.
Amino Acid Sequence↗
CD5 expression by B lymphocytes and its regulation upon Epstein-Barr virus transformation.
Dim expression of CD5 on human B lymphocytes has been used to delineate B1 and B2 subsets. Nevertheless, others have suggested that the molecule is an activation marker and does not predicate a subset distinction. We have used enzymatic amplification staining, a technology that enhances the resolution of flow cytometric analysis of cell surface molecules by as much as 100-fold, to determine that essentially all human B cells express CD5. Furthermore, we show that this expression is regulated during Epstein-Barr virus transformation.
B-Lymphocytes↗
PKA, PKC, and AKAP localization in and around the neuromuscular junction.
BACKGROUND: One mechanism that directs the action of the second messengers, cAMP and diacylglycerol, is the compartmentalization of protein kinase A (PKA) and protein kinase C (PKC). A-kinase anchoring proteins (AKAPs) can recruit both enzymes to specific subcellular locations via interactions with the various isoforms of each family of kinases. We found previously that a new class of AKAPs, dual-specific AKAPs, denoted D-AKAP1 and D-AKAP2, bind to RIalpha in addition to the RII subunits. RESULTS: Immunohistochemistry and confocal microscopy were used here to determine that D-AKAP1 colocalizes with RIalpha at the postsynaptic membrane of the vertebrate neuromuscular junction (NMJ) and the adjacent muscle, but not in the presynaptic region. The labeling pattern for RIalpha and D-AKAP1 overlapped with mitochondrial staining in the muscle fibers, consistent with our previous work showing D-AKAP1 association with mitochondria in cultured cells. The immunoreactivity of D-AKAP2 was distinct from that of D-AKAP1. We also report here that even though the PKA type II subunits (RIIalpha and RIIbeta) are localized at the NMJ, their patterns are distinctive and differ from the other R and D-AKAP patterns examined. PKCbeta appeared to colocalize with the AKAP, gravin, at the postsynaptic membrane. CONCLUSIONS: The kinases and AKAPs investigated have distinct patterns of colocalization, which suggest a complex arrangement of signaling micro-environments. Because the labeling patterns for RIalpha and D-AKAP 1 are similar in the muscle fibers and at the postsynaptic membrane, it may be that this AKAP anchors RIalpha in these regions. Likewise, gravin may be an anchor of PKCbeta at the NMJ.
A Kinase Anchor Proteins↗
HIV protease inhibitor substitution in patients with lipodystrophy: a randomized, controlled, open-label, multicentre study.
BACKGROUND: Lipodystrophy, dyslipidaemia and insulin resistance often complicate protease inhibitor-containing antiretroviral therapy. The aims of this study were to determine if these are reversible with continued HIV suppression following protease inhibitor substitution. METHODS: Eighty-one HIV protease inhibitor recipients (78 male; mean antiretroviral therapy, 55 months) with predominant peripheral lipoatrophy, HIV RNA < 400 copies/ml plasma for at least the preceding 6 months, and no prior abacavir, non-nucleoside analogue or adefovir therapy were randomized 3 : 2 to continue nucleoside analogues and substitute protease inhibitor(s) with abacavir, nevirapine, adefovir and hydroxyurea (n = 49) or to continue all therapy (n = 32) with an option to switch at week 24. The primary endpoints were total body fat and HIV RNA at week 24. Other assessments were regimen safety, regional body composition, metabolic parameters, quality of life, and CD4 T-lymphocyte counts to week 48. RESULTS: There was a greater decline in total body fat in the switch group than in the continue group (-1.6 and -0.4 kg, respectively at week 24; P = 0.006). This comprised greater declines in limb and subcutaneous abdominal fat, and in intra-abdominal fat of patients with moderate or severe abdominal fat accumulation. Viral suppression was similar, despite 18 (37%) switch group patients ceasing at least one study drug by week 24 because of adverse events. Total cholesterol and triglycerides declined more in the switch group (both P < 0.002). High density lipoprotein cholesterol increased significantly in both groups at week 48 (P < 0.02). There was no change for any glycaemic parameter. CONCLUSIONS: In predominantly lipoatrophic patients, switching from HIV protease inhibitor therapy lead to improved lipids and less intra-abdominal fat, but also to less peripheral fat, and had minimal effect on insulin resistance. Virological control in these heavily pretreated patients was unaffected, despite frequent switch drug cessations.
Adenine↗
Plasmid vaccination with insulin B chain prevents autoimmune diabetes in nonobese diabetic mice.
The insulin B (InsB) chain bears major type 1 diabetes-associated epitopes of significance for disease in humans and nonobese diabetic (NOD) mice. Somatic expression of InsB chain initiated early in life by plasmid inoculation resulted in substantial protection of female NOD mice against disease. This was associated with a T2 shift in spleen, expansion of IL-4-producing and, to a lesser extent, of IFN-gamma-secreting T cells in pancreatic lymph nodes, as well as intermolecular Th2 epitope spreading to glutamic acid decarboxylase determinants. A critical role of IL-4 for the Ag-specific protective effect triggered by plasmid administration was revealed in female IL-4(-/-) NOD mice that developed diabetes and higher Th1 responses. Coadministration of IL-4-expressing plasmid or extension of the vaccination schedule corrected the unfavorable response of male NOD mice to DNA vaccination with InsB chain. Thus, plasmid-mediated expression of the InsB chain early in diabetes-prone mice has the potential to prevent transition to full-blown disease depending on the presence of IL-4.
Animals↗
Age-related impairment of synaptic transmission but normal long-term potentiation in transgenic mice that overexpress the human APP695SWE mutant form of amyloid precursor protein.
We have studied synaptic function in a transgenic mouse strain relevant to Alzheimer's disease (AD), overexpressing the 695 amino acid isoform of human amyloid precursor protein with K670N and M671L mutations (APP(695)SWE mice), which is associated with early-onset familial AD. Aged-transgenic mice had substantially elevated levels of Abeta (up to 22 micromol/gm) and displayed characteristic Abeta plaques. Hippocampal slices from 12-month-old APP(695)SWE transgenic animals displayed reduced levels of synaptic transmission in the CA1 region when compared with wild-type littermate controls. Inclusion of the ionotropic glutamate receptor antagonist kynurenate during preparation of brain slices abolished this deficit. At 18 months of age, a selective deficit in basal synaptic transmission was observed in the CA1 region despite treatment with kynurenate. Paired-pulse facilitation and long-term potentiation (LTP) were normal in APP(695)SWE transgenic mice at both 12 and 18 months of age. Thus, although aged APP(695)SWE transgenic mice have greatly elevated levels of Abeta protein, increased numbers of plaques, and reduced basal synaptic transmission, LTP can still be induced and expressed normally. We conclude that increased susceptibility to excitotoxicity rather than a specific effect on LTP is the primary cause of cognitive deficits in APP(695)SWE mice.
Aging↗
Oral lesions in sheep and cattle in Dumfries and Galloway.
Explore the source record for details and available documents.
Animals↗
High-throughput metabolism: making the most of the technology.
Explore the source record for details and available documents.
Journal Article↗