[The effects and limitations of myofunctional therapy].
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Biomedical subjects
Publications and source records attributed to T Ohno.
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The complete nucleotide sequence of TMV RNA (common strain) reported in [Proc. Natl. Acad. Sci. USA (1982) 79, 5818] its 5'-end to be represented by two variants which differed in length. We have tested that result and sequenced the 5'-terminal regions of two strains of TMV RNA (common strain OM and tomato strain L) using cloned cDNA copies. The results showed that the 5'-terminal region of the TMV genome is not polymorphic and that one of the two variants cited above represents a tomato strain but not the common strain.
The complete cDNA of hop stunt viroid (HSV) has been cloned by the method of Okayama and Berg (Mol.Cell.Biol.2,161-170. (1982] and the complete nucleotide sequence has been established. The covalently closed circular single-stranded HSV RNA consists of 297 nucleotides. The secondary structure predicted for HSV contains 67% of its residues base-paired. The native HSV can possess an extended rod-like structure characteristic of viroids previously established. The central region of the native HSV has a similar structure to the conserved region found in all viroids sequenced so far except for avocado sunblotch viroid. The sequence homologous to the 5'-end of U1a RNA is also found in the sequence of HSV but not in the central conserved region.
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The cDNA copies of tobacco mosaic virus (TMV)-tomato strain (L) genome were cloned by the method of Okayama and Berg (Mol. Cell. Biol. 2, 161-170. (1982)) and the sequence of 1,614 nucleotides at the 3' end was determined. The sequence encompasses the 30K and the coat protein cistron which are located in residues 685-1, 479 and 203-682 from the 3' end of the genome respectively. The close relationship between the tomato and the common strain was shown on the level of the nucleotide sequence. Highly homologous regions are found in the 3' non-coding region, the assembly origin and the 5' flanking region of the 30K protein cistron. The comparison of the deduced amino acid sequence between the tomato and the common strain shows that the 30K protein is composed of the conserved N-terminal four-fifth and the highly divergent region near the C-terminus.
Horse kidney Na,K-ATPase solubilized with dodecyloctaethyleneglycolether was subjected to high performance gel chromatography (HPLC) on TSK G 4000 SW in the presence of 0.01% C12E8. Successive on-line measurements of low angle laser light scattering (LS) and refractive index (RI) were made, and values of refractive index increment (dn/dc) were measured with the same differential refractometer under the same conditions. Two peaks (peak-2 and peak-3) with Na,K-ATPase activity besides that at the void volume were detected in the HPLC effluent fractions, and from their (dn/dc) values and the linear plot of protein standards, the M.W.s of these peaks were calculated to be roughly 535K and 175K respectively. Both peaks showed alpha and beta, but not gamma, bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The activity peak fractions obtained after glycerol density gradient centrifugation to remove detergent micelles showed HPLC peak-2 and peak-3. The ratios of (Output) LS/(Output)RI of peak-2 to that of peak-3 and (Output) LS/(Output)UV 280 of the peak-2 to that of peak-3 were both nearly 3. The above findings and the results of electron microscopy of findings and the results of electron microscopy of negatively stained preparations strongly suggested that peak-2 and peak-3 of enzyme activity consist of the trimer (alpha beta)3 and the monomer (alpha beta), respectively.
Adrenal demedullation combined with chemical sympathectomy with 6-hydroxydopamine (ACS) lowered plasma glucagon and insulin levels in rats. Acute cold exposure increased plasma glucagon in both ACS and control rats, while it increased plasma insulin only in ACS rats. ACS rats responded to cold with a smaller increase in plasma glycerol and a more pronounced elevation of plasma free fatty acids.
Our previous study provided a new 2-dimensional echocardiographic technique to detect peripheral right coronary artery aneurysms in Kawasaki disease, with use of the subcostal approach. An additional study was performed to detect peripheral left coronary artery aneurysms. Because the left anterior descending artery runs along the anterior interventricular sulcus and the left circumflex artery along the mitral valve ring, these regions were searched for coronary aneurysms by use of the subcostal imaging approach. Among 143 patients with Kawasaki disease, 44 left coronary aneurysms were visualized in 22 patients. Three aneurysms at the origin of the obtuse marginal artery and 1 in the further peripheral site of the left circumflex artery were observed in 3 patients. Two aneurysms at the origin of the second diagonal branch of the peripheral left anterior descending artery were detected. These echocardiographic studies were done prospectively, and their features coincided well in size, shape, and anatomic location with confirmatory angiographic appearances.
Involvement of the tobacco mosaic virus (TMV) coded 30K protein in a virus transport function within the infected plant has been suggested. Previously a temperature sensitive mutant, TMV Ls 1, that is defective in cell-to-cell movement at a restrictive temperature, was reported. To demonstrate a relationship between the 30K protein and the transport function, the nucleotide sequences of the 30K and coat protein cistrons of the mutant, TMV Ls 1, and the wild type, TMV L (tomato strain) were compared. A single base substitution which causes replacement of a proline codon in the L strain by a serine codon was found in the Ls 1 mutant. Results support the notion that the 30K protein is responsible for the virus transport function.
We examined the metabolites of o-phenylphenol (OPP) in the urine of male and female rats dosed with 2% sodium o-phenylphenate (OPP-Na) in food from the age of 5 wk for 136 days. The urinary metabolites of OPP-Na produced during the 24 hr after OPP-Na feeding accounted for 55% of the dose in male rats and 40% in females. The main metabolites were OPP-glucuronide and 2,5-dihydroxybiphenyl (2,5-DHBP)-glucuronide. OPP metabolites in the free form accounted for only 1% of the total phenolic metabolites excreted. 2,5-DHBP was rapidly converted to the corresponding quinone in aqueous solvents but not in organic solvents. There was a clear sex difference in the proportions of urinary metabolites; the amount of 2,5-DHBP excreted by male rats in 24-hr urine was more than seven times that excreted by females. This result may be related to the finding that bladder tumours occur in male but not female rats fed OPP-Na in the diet.
Human skin explants obtained from 2- to 5-yr-old patients with harelips were cultured in NCTC 168 medium at 37 degrees C, in a humidified atmosphere containing 5% CO2 in air. After a 2-week incubation period, the newly grown cells were studied with special reference to tight junctions by freeze-fracture electron microscopy. Many completely formed tight junctions were observed between the uppermost living cells of migrating epithelium, and fragmented tight junctions were seen between the lower layer cells. The tight junctions in the uppermost cells developed so well that they formed a belt-like network consisting of two to six rows of strands. This observation may suggest that human keratinocytes have the ability to produce tight junctions perfectly enough to serve as a barrier, although no complete tight junctions were formed in situ.
An autopsy case of pseudomesotheliomatous carcinoma of the right lung in a 56-year-old man occupationally exposed to stone dust is presented. From open biopsy specimens in which polyhedral epithelium-like cells appeared arranged in nests, sheets, and trabecula without apparent tubular pattern, a malignant pleural mesothelioma was suspected. At autopsy, the right pleural cavity was obliterated by the tumor mass which covered the collapsed pulmonary parenchyma and was clearly demarcated from it. The gross appearance of the tumor was similar to that of malignant pleural mesothelioma. Histologically, marked interstitial fibrosis of the subpleural parenchyma of both lungs was observed, and the tumor tissue was interspersed in the parenchyma adjacent to the tumor mass. The tumor showed both intracytoplasmic and intercellular positive materials with colloidal iron, alcian blue (pH 2.5), and toluidine blue stains, which entirely disappeared after streptomyces hyaluronidase digestion. A small amount of intracytoplasmic PAS-positive material resistant to diastase digestion was also observed. Immunohistochemical staining for carcinoembryonic antigen, which is said to be negative in malignant pleural mesothelioma, was positive intracellularly. There were no histologic findings indicating asbestos exposure. From these findings, the authors made a diagnosis of poorly differentiated adenocarcinoma, which was characterized by the production of hyaluronic acid.
The topographical distribution of interneurones mediating disynaptic inhibition of dorsal spinocerebellar tract (d.s.c.t.) cells from group I muscle afferents in the cat was investigated using both physiological and morphological techniques. Lesions of either the dorsal funiculi or of the lateral and ventral funiculi were made between L4 and L5 segments in two groups of cats. I.p.s.p.s. evoked from group I afferents were seen after both these lesions, showing that the i.p.s.p.s were evoked by interneurones located more caudally as well as by interneurones in the same segments as Clarke's column. Distribution of the caudally located interneurones in the lower lumbar segments was investigated after marking these interneurones with horseradish peroxidase retrogradely transported from Clarke's column. The horseradish peroxidase was injected along L3-L4 segments of Clarke's column in two cats with transected dorsal funiculi. The marked cells were found in L5, L6, L7 and S1 segments, with a highest density in L6 and L7. They were seen in laminae V, VI and VII. A search was made for interneurones which could be antidromically invaded following stimuli applied in Clarke's column and were monosynaptically excited by group I afferents. Such interneurones were found at locations corresponding to laminae V-VI of Rexed. The latencies of antidromic and orthodromic responses were within ranges allowing them to mediate disynaptic inhibition of d.s.c.t. cells.
The aim of the study was to investigate whether inhibition of dorsal spinocerebellar tract (d.s.c.t.) cells evoked from group I afferents is mediated by the same interneurones which mediate the non-reciprocal inhibition of hind-limb motoneurones. The origin of inhibition of d.s.c.t. cells from group I afferents was compared in intact preparations, after lesions of the dorsal funiculi (when it could only be mediated by lower lumbar interneurones) and after lesions of the lateral and ventral funiculi (when it would be expected to be evoked by upper lumbar interneurones). In all three preparations extensors were the most common source of inhibition, as in motoneurones. Lower lumbar interneurones inhibiting d.s.c.t. cells were found to be co-excited by group I (Ia and/or Ib) and cutaneous and joint afferents, and by rubrospinal tract fibres, as are interneurones mediating inhibition of motoneurones. Co-excitation by group I and rubrospinal fibres was also found for upper lumbar interneurones. I.p.s.p.s were evoked in hind-limb motoneurones from within Clarke's column in cats with the dorsal funiculi cut between L4 and L5 segments; they were evoked at thresholds as low as 2 microA, i.e. by stimuli with very local actions. The latencies of these i.p.s.p.s were short enough to allow them to be evoked monosynaptically via axonal branches of the same interneurones which projected to Clarke's column. Correspondingly, i.p.s.p.s were evoked in d.s.c.t. cells from within motor nuclei in L7 segments; they were evoked at similarly low thresholds and with similar latencies. In confirmation of previous reports i.p.s.p.s of Ia origin evoked in d.s.c.t. cells were not found to be depressed by Renshaw cells, which excludes their mediation by interneurones responsible for Ia reciprocal inhibition. The study leads to the conclusion that the inhibition of d.s.c.t. cells from group I afferents is, at least in part, collateral to the non-reciprocal inhibition of lumbar motoneurones.
Postural abnormalities in a 66-year-old woman with ataxic myxoedema were recorded by stabilography during treatment. After replacement therapy, 3 Hz power in the sagittal plane was greater than before treatment; this peak shifted to a higher frequency and finally its ratio became normal. This is the first report of recovery of a 3 Hz body oscillation in ataxic myxoedema.
Leukocyte components which can inhibit or enhance the uptake of 3H-thymidine by peripheral lymphocytes can be prepared by ultrafiltration of leukocyte extracts followed by gel filtration on Sephadex G-10. A potent inhibitor of lymphocyte transformation is present in an ultrafiltrate fraction having a nominal molecular weight of 5,000-10,000. Three other components having inhibitory, and one component having enhancing, activity on lymphocyte transformation were found in an ultrafiltrate fraction having a molecular weight less than 5,000. The inhibitory components in the latter were principally derived from polymorphs and inhibited both T and B lymphocyte proliferation. Enhancing components were mainly derived from mononuclear cells and augmented the proliferation of T cells only.
A granulocyte/macrophage colony-stimulating factor (Peak-1 CSF) was partially purified from the medium of a serum-free culture of Yoshida sarcoma cells (Line YSSF-212T). Its elution position in gel-filtration chromatography corresponded to a molecular weight of about 22,000. The factor had an isoelectric point at pH 4.5 and a sedimentation coefficient of 2.3 S. The major part of its activity was not bound by Concanavalin A-Sepharose. Although CSF activity behaved as a single component in the gel-filtration and isoelectrofocussing procedures, subsequently it was resolved into two species by preparative discontinuous polyacrylamide gel-electrophoresis. This resolution indicates microheterogeneity of the CSF molecule. Oxidation with periodate readily inactivated L . P3-cell CSF, but the YSSF-cell CSF was fairly resistant. Moreover, titration with anti-L cell CSF serum showed a definite difference between L . P3-cell CSF and YSSF-cell CSF.
Serum-free culture is a useful method for identifying the growth requirements of normal cells of human origin and comparing them to the requirements of transformed human cells. We examined differential growth requirements of serum factors (bovine serum albumin (BSA) and low density lipoprotein (LDL) on normal (HEL), Co-60 gamma ray-transformed (CT-1) and 4-nitroquinoline 1-oxide-transformed (SUSM-1) human fibroblast cells in the absence of serum. A decreased requirement for serum was closely related to a decreased requirement for BSA in both types of transformed cells. The growth rate and saturation density of HEL and CT-1 cells grown in serum-free medium supplemented with the optimal concentrations of BSA were almost equal to the rates of cells grown in serum-supplemented medium. When BSA alone was added to the serum-free medium it did not support the continuous growth of SUSM-1 cells because of cellular detachment. An addition of LDL to serum-free cultures not only promoted the moderate growth of HEL cells, it promoted the continuous growth of SUSM-1 cells as well. In contrast, LDL was not required for the optimal growth of CT-1 cells. Transformed human cells specifically had reduced quantitative and qualitative growth requirements for one or more serum factors.