Low prevalence of thyrotropin receptor antibody in primary hypothyroidism in Japan

The prevalence of TSH receptor antibody (TRAb), measured by RRA assay (TSH-binding inhibitor immunoglobulin, TBII) and biological stimulation-blocking assay (thyroid-stimulation blocking antibody, TSBAb), was examined in 134 consecutive patients with primary hypothyroidism due to autoimmune thyroiditis [83 patients with goitrous Hashimoto's disease (group A) and 51 with primary atrophic hypothyroidism (group B)]. In group A, TBII was detected in 6 patients (7%), TSBAb in 7 (8%), and both in 4 (5%). Similarly, in group B, TBII was detected in 7 patients (14%), TSBAb in 7 (14%), and both in 5 (10%). TBII with TSBAb activity was low or moderate in group A, but strongly positive in group B. No relationship was apparent in either group between TBII or TSBAb activity and any clinical or laboratory parameter examined. Moreover, no clinical or laboratory findings distinguished patients with TRAb from those without. The low prevalence of TRAb in primary hypothyroidism suggests that intrathyroidal cell-mediated destructive mechanisms may be more important in the pathogenesis of hypothyroidism in autoimmune thyroiditis.     

Simultaneous measurement of TSH-binding inhibitor immunoglobulin and thyroid stimulating autoantibody activities using cultured FRTL-5 cells in patients with untreated Graves' disease

A new and practical assay was developed using cultured FRTL-5 cells for simultaneous assessment of TS-ab and TSH-binding inhibitor immunoglobulin, allowing direct comparison of these two activities under the same conditions. Subsequent to the TS-ab assay in which extracellular cAMP concentration in Hanks' medium without NaCl was determined, [125I]b TSH in this medium was added to observe the ability of serum Ig to inhibit the binding of [125I]b TSH to FRTL-5 cells. We found a much higher specific binding of [125I]b TSH to FRTL-5 cells and a much greater inhibition of [125I]bTSH binding to the cells exposed to Graves' Ig in hypotonic NaCl-free than in NaCl containing Hanks' medium, indicating that the binding of both TSH and Graves' Ig to the TSH receptor was salt-sensitive. The inhibitory activity of [125I]bTSH binding to the cells was 0.2 +/- 4.6% (mean +/- SD) in 45 normals. Inter-assay coefficients of variation in two positive controls with the mean values of 18.0 and 65.8% were 15.8 and 16.5%, respectively. Among 46 patients with untreated hyperthyroidism owing to Graves' disease, 45 (97.8%) were positive for TS-ab; 35 (76.1%) and 40 (87.0%) were positive for TSH-binding inhibitor in Ig assays using FRTL-5 cells and solubilized porcine thyroid membranes, respectively. TS-ab activities correlated less closely with TSH-binding inhibitory activities determined using FRTL-5 cells (r = 0.576, p less than 0.001) than with those determined using porcine thyroid membranes (r = 0.745, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular characterization of naturally occurring glycoprotein C-negative herpes simplex virus type 1

Summary We previously isolated glycoprotein C (gC)-negative herpes simplex virus type 1 (HSV-1) mutants, TN-1, TN-2 and TN-3, from a patient with recurrent herpetic keratitis at one-year intervals. In the present study, the molecular basis for the inability of these clinical isolates to express gC was examined. The nucleotide sequence of the gC gene of the TN-1 strain was compared with that of the HSV-1 KOS strain. In the open reading frame of the gC gene, there were 12 nucleotide differences between the TN-1 and KOS strains, seven of which led to amino acid substitutions. Importantly, one of them was the codon change from CAG for glutamine at position 280 to TAG for the amber termination codon. Accordingly, the TN-1 strain produced a truncated gC with a predicted molecular weight, which was secreted into the extracellular fluid. These results suggest that this amber mutation in the TN-gC gene results in a premature termination of gC translation and is the cause of the gC-negative phenotype of the TN strains. It is expected that these extremely rare HSV-1 strains will provide us with valuable information concerning the in vivo functions of gC, especially in ocular diseases.     

Construction of radiation-reduced hybrids and their use in mapping of microclones from chromosome 10p11.2-q11.2

Summary Radiation-reduced hybrids for mapping of DNA markers in the pericentromeric region of chromosome 10 were developed. A Chinese hamster/human somatic cell hybrid (762-8A) carrying chromosomes 10 and Y as the only human material were exposed to 40,000 rads of irradiation and then rescued by fusion with non-irradiated recipient Chinese hamster cells (GM459). Southern hybridization analyses revealed that 10 of 128 HAT-resistant clones contained human chromosomal fragments corresponding to at least one marker locus betweenFNRB (10p-11.2) andRBP3 (10q11.2). These hybrids were then used to map microdissection clones previously isolated and roughly mapped to this chromosomal region by fluorescencein situ hybridization (FISH). Two of the six microclones studied could be mapped to the proximity of the D10-S102 locus. These radiation hybrids are useful for the construction of refined genetic maps of the pericentromeric region of chromosome 10.     

The effects of pacing-induced left bundle branch block on left ventricular systolic and diastolic performances

To assess the effects of pacing-induced left bundle branch block on left ventricular (LV) systolic and diastolic performance, we performed digital subtraction ventriculography while simultaneously measuring LV pressure with a catheter tip micromanometer. The subjects included 10 patients with a sinus rhythm, a normal QRS duration and PR interval within 0.22 sec. LV performance was assessed during both right atrial pacing (AP) and atrioventricular sequential pacing (AVP) at the same pacing rate. The atrioventricular pacing interval during AVP was adjusted to be the maximal interval that showed the QRS configuration seen during complete right ventricular pacing. LV end-diastolic pressure and volume during AVP did not differ from those during AP. Peak positive and negative dp/dt during AVP were significantly lower than those during AP. Time constants were also significantly longer during AVP. The QRS duration during AVP significantly correlated with end-systolic volume and time constants, and inversely correlated with ejection fraction and +dp/dt. These observations indicated that conduction disturbance per se, induced by AVP, could not only impair LV systolic performance but also diastolic performance, possibly due to asynchronous contraction and relaxation of the left ventricle.     

Cell culture studies of a patient with congenital lipoatrophic diabetes—Normal insulin binding with alterations in intracellular glucose metabolism and insulin action

Insulin binding and the action of insulin on several aspects of glucose metabolism have been investigated in cultured fibroblasts derived from a patient with congenital lipoatrophic diabetes and compared to cultures from 9 nondiabetic controls. Incorporation of glucose was elevated in the patient's cells at glucose levels above 0.1 mM. When distribution of labelled glucose was examined, cell associated glycogen and acid soluble material were increased, but the greatest increment was in lactate production. Insulin binding, as indicated by maximum specific ¹²⁵I-insulin binding and concentration of unlabelled insulin for 50% displacement, was normal, although insulin regulation of the insulin receptor was diminished. Insulin stimulation of total glucose incorporation was reduced in cells from the patient. When insulin stimulation of glycogen synthase was measured directly, the response to insulin was also attenuated. On the other hand, insulin stimulation of hexose transport appeared to be unimpaired. The data indicate alterations in both cell glucose metabolism and insulin response which may be related to the observed insulin resistance of this disorder.     

N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, a calmodulin antagonist, inhibits cell proliferation.

N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) and its derivatives are putative calmodulin antagonists that bind to calmodulin and inhibit Ca2+/calmodulin-regulated enzyme activities. Autoradiographic studies using tritiated W-7 showed that this compound penetrates the cell membrane, is distributed mainly in the cytoplasm, and inhibits proliferation of Chinese hamster ovary K1 (CHO-K1) cells. Cytoplasmic [3H]W-7 was excluded completely within 6 hr after removal of [3H]W-7 from the culture medium. N-(6-aminohexyl)-1-naphthalenesulfonamide, an analogue of W-7 that interacts only weakly with calmodulin, proved to be a much weaker inhibitor of cell proliferation. CHO-K1 cells were synchronized by shaking during mitosis and then released into the cell cycle in the presence of 25 microM W-7 or 2.5 mM thymidine for 12 hr. Cell division was observed approximately 6 hr later. The results suggest that the effect of W-7 on cell proliferation might be through selective inhibition of the G1/S boundary phase, which is similar to the effect of excess thymidine. This pharmacological demonstration that cytoplasmic calmodulin is involved in cell proliferation is significant; W-7 and its derivatives may be useful tools for research on calmodulin and cell biology-related studies.     

Effects of the Implant Design on Peri‐Implant Bone Stress and Abutment Micromovement: Three‐Dimensional Finite Element Analysis of Original Computer‐Aided Design Models

Background: Occlusal overloading causes peri‐implant bone resorption. Previous studies examined stress distribution in alveolar bone around commercial implants using three‐dimensional (3D) finite element analysis. However, the commercial implants contained some different designs. The purpose of this study is to reveal the effect of the target design on peri‐implant bone stress and abutment micromovement. Methods: Six 3D implant models were created for different implant–abutment joints: 1) internal joint model (IM); 2) external joint model (EM); 3) straight abutment (SA) shape; 4) tapered abutment (TA) shapes; 5) platform switching (PS) in the IM; and 6) modified TA neck design (reverse conical neck [RN]). A static load of 100 N was applied to the basal ridge surface of the abutment at a 45‐degree oblique angle to the long axis of the implant. Both stress distribution in peri‐implant bone and abutment micromovement in the SA and TA models were analyzed. Results: Compressive stress concentrated on labial cortical bone and tensile stress on the palatal side in the EM and on the labial side in the IM. There was no difference in maximum principal stress distribution for SA and TA models. Tensile stress concentration was not apparent on labial cortical bone in the PS model (versus IM). Maximum principal stress concentrated more on peri‐implant bone in the RN than in the TA model. The TA model exhibited less abutment micromovement than the SA model. Conclusion: This study reveals the effects of the design of specific components on peri‐implant bone stress and abutment displacement after implant‐supported single restoration in the anterior maxilla.