New epitopes and function of anti-M3 muscarinic acetylcholine receptor antibodies in patients with Sjögren's syndrome.

M3 muscarinic acetylcholine receptor (M3R) plays a crucial role in the secretion of saliva from salivary glands. It is reported that some patients with Sjögren's syndrome (SS) carried inhibitory autoantibodies against M3R. The purpose of this study is to clarify the epitopes and function of anti-M3R antibodies in SS. We synthesized peptides encoding the extracellular domains of human-M3R including the N-terminal region and the first, second and third extracellular loops. Antibodies against these regions were examined by enzyme-linked immunosorbent assay in sera from 42 SS and 42 healthy controls. For functional analysis, human salivary gland (HSG) cells were preincubated with immunoglobulin G (IgG) separated from sera of anti-M3R antibody-positive SS, -negative SS and controls for 12 h. After loading with Fluo-3, HSG cells were stimulated with cevimeline hydrochloride, and intracellular Ca(2+) concentrations [(Ca(2+) )i] were measured. Antibodies to the N-terminal, first, second and third loops were detected in 42·9% (18 of 42), 47·6% (20 of 42), 54·8% (23 of 42) and 45·2% (19 of 42) of SS, while in 4·8% (two of 42), 7·1% (three of 42), 2·4% (one of 42) and 2·4% (one of 42) of controls, respectively. Antibodies to the second loop positive SS-IgG inhibited the increase of (Ca(2+) )i induced by cevimeline hydrochloride. Antibodies to the N-terminal positive SS-IgG and antibodies to the first loop positive SS-IgG enhanced it, while antibodies to the third loop positive SS-IgG showed no effect on (Ca(2+) )i as well as anti-M3R antibody-negative SS-IgG. Our results indicated the presence of several B cell epitopes on M3R in SS. The influence of anti-M3R antibodies on salivary secretion might differ based on these epitopes.

Physical mapping of the conjugative bacteriocin plasmid pPD1 of Enterococcus faecalis and identification of the determinant related to the pheromone response.

The pheromone-responding conjugative bacteriocin plasmid pPD1 (59 kb) of Enterococcus faecalis was mapped physically by using a relational clone approach, and transposon analysis with Tn917 (Emr) or Tn916 (Tcr) facilitated the location of the bacteriocin-related genes in a segment of about 6.7 kb. Tn917 insertions within a 3-kb region resulted in constitutive clumping. The nucleotide sequence of the region that included the insertions giving rise to constitutive clumping was determined. The region of pPD1 spanned about 8 kb and was found to contain a number of open reading frames, some of which were named on the basis of homologies with two other pheromone-responding plasmids, pAD1 and pCF10. The genes were arranged in the sequence repB-repA-traC-traB-traA-ipd-traE-traF- orfY-sea-1 with all but repB and traA oriented in the same (left-to-right) direction. traC and traB corresponded, respectively, to traC and traB of pAD1 and to prgY and prgZ of pCF10.

The configuration of the laminas and facet joints in degenerative spondylolisthesis. A clinicoradiologic study.

The configuratory variation of laminas and facet joints was discussed with reference to development of degenerative spondylolisthesis and its clinical symptoms. The authors have classified the configuration of laminas in the lower lumbar spine into three different types, (WI, W2, and N), based on the two characteristic features of the lamina: 1) whether or not the inferior articular processes are wider than the waist part of the lamina, and 2) whether or not the facet joint spaces can be recognized on plain anteroposterior (AP) radiographs. Types W1 and W2 laminas have the laterally prominent inferior articular processes, but the facet joint spaces are not visible on AP view in Type W1. Type N lamina has narrow inferior articular processes and its facet joint spaces can be recognized on AP view. Computed tomography (CT) scanning revealed that the direction of facet joints of Types W2 and N laminas was more sagittal than that of Type W1, and that all of Type N laminas with the narrow inferior articular processes have a sagittal facet. Type N lamina was rarely seen in the 257 controls, but was quite frequent in degenerative spondylolisthesis, 3.5% and 45.8%, respectively. Patients with Type N laminas were likely to show more severe clinical symptoms such as difficulty walking and neurologic deficits.(ABSTRACT TRUNCATED AT 250 WORDS)

Histomorphometric study of trabecular channels in normal iliac bone.

Cancellous bone channels in the normal iliac crest have been studied histologically and by histomorphometry, and their biological role has been considered. Eighty percent of trabecular channels were typical osteons with the same structural and remodeling features as cortical osteons. The similarity of osteons in these two locations was corroborated by the comparability of morphometric features. The points of difference between osteons in the two types of bone were irregular configurations of trabecular osteons and marrow cells in the central canal in some. Since the number of trabecular osteons decreases with age, and since active trabecular resorption cavities were few in number, it is unlikely that additional osteons are formed in normal bone after the active modeling phase of bone growth. It is improbable that they make a significant contribution to bone remodeling since their number decreases with age, and since the available surface of trabecular channels for remodeling is extremely small. However, it is probable that they aid the intraosseous microcirculation and mineral exchange in thick trabecula and bifurcations, where they are mainly located. The demarcation curves at the 95% confidence interval, which suggest the normal range for numbers of channels, was calculated from the scatter diagram against bone area.

[Concentration of cefotaxime and its metabolite in human bone marrow blood].

Cefotaxime (CTX) was intravenously administered in an amount of 2.0 g to each of 34 adult patients before the surgery mainly of the hip joint. Samples of the blood from the bone marrow around the trochanter were taken at the time of the operation. At the same time blood samples were taken from peripheral veins. The sample was centrifuged and the supernatant was analyzed for CTX and desacetyl-CTX. The concentration of CTX in the marrow blood was 150.9 micrograms/ml and that in the blood was 182.5 micrograms/ml in the earliest samples taken at 20 minutes after injection. In the 44 pairs of samples, the concentration of CTX in the marrow blood was lower than that in the peripheral blood in all the cases except 4. The concentration of desacetyl-CTX (Des-CTX), however, in the marrow blood was higher than in the peripheral blood in 33 of the 44 pairs of specimens. Since the degradation of the drug progresses with time, the ratio of Des-CTX to CTX increased with time. This trend was particularly marked in the bone marrow blood and can be expressed as Y = 113.0 + 0.32 t, when Y is the ratio percentage (Des-CTX/CTX) and t is time after the injection of the drug in minute. Thus, CTX transferred into the bone marrow tends to remain there and transformed into the desacetyl form.

The functional significance of human notochord in the development of vertebral column. An electron microscopic study.

Twenty-one human embryos or fetuses from four to twenty-one weeks in gestation age were submitted to light and electron microscopic observations of the notochord and its related structures. The notochord cells were found to have densely developed rER and Golgi areas, which were correlated with an activity to secrete into their surroundings specific ground substance probably containing proteoglycan. The highest differentiation of this cell as estimated by the peak secretory activity was reached at about ten weeks, while signs of degeneration emerged shortly thereafter and became increasingly apparent with the advancing gestation. The synthesis and secretion of matrices were likely to be taken over by the fibrocartilage cells surrounding the notochord by about 20 weeks when the notochord cells had undergone severe degenerative changes. Another characteristic feature of the notochord cells at their fully differentiated stage was abundance in intracytoplasmic glycogen that coexisted with striking paucity of mitochondria. This was interpreted as suggesting the metabolic dependence of these cells upon anaerobic glycolysis even at the peak of their secretory performance.

Change of prostaglandin E level in joint fluids after treatment with flurbiprofen in patients with rheumatoid arthritis and osteoarthritis.

The prostaglandin E (PGE) level in the knee joint fluid was determined by radioimmunoassay before and after anti-inflammatory therapy with flurbiprofen in 8 patients with rheumatoid arthritis (RA) and 4 patients with osteoarthritis (OA). The level of PGE in RA joint fluids before the anti-inflammatory treatment was 9.5-1.2 ng/ml and in proportion with the leucocyte count in the joint fluid. A marked decrease of the PGE level was attained with flurbiprofen treatment in 4 patients whose initial PGE levels had been higher than 3.2 ng/ml, while 4 patients with lower PGE levels, namely, 1.4-1.2 ng/ml, did not respond to the drug treatment. In all the OA patients the PGE levels was no higher than 1.5 ng/ml and refractory to the anti-inflammatory therapy.

Lipid composition of adipose tissue from "membranous lipodystrophy".

The lipid composition of the adipose tissue from the bone marrow and that of the subcutaneous depot fat of four patients of membranous lipodystrophy were investigated. It was found that the lipid composition of the samples from the cases of membranous lipodystrophy was almost similar to that in controls. Also discussions were made on mechanism related to the accumulation of abnormal depot fat.

Excretion of glucose-containing oligosaccharides in urines of orthopedic patients.

Urinary neutral oligosaccharides of various connective tissue diseases were studied by gel-filtration through Sephadex G--10 after treatments with cetylpyridinium chloride (CPC), Dowex 50 (H+ form) and Dowex 1 (Cl- form), in succession. Increased excretion of urinary glucose-containing oligosaccharides, specifically glucosylgalactose was observed in most of the patients with chondrosarcoma, rheumatoid arthritis, Werner's syndrome, Rothmund Thomson syndrome and Morquio's disease. However, urinary excretion of neutral oligosaccharides in the patients with osteosarcoma and other tumorous conditions, and some systemic disorders in the connective tissues, examined in the present study, showed almost normal values. It is indicated, therefore, that the activity of glucosidase in insufficient for the glucose-containing oligosaccharides produced from the ground substance(s) in the former type connective tissue diseases.

Comparison of roentgenological and histological features of bone changes in primary hyperparathyroidism.

Fifteen cases of bone disease in primary hyperparathyroidism were investigated by comparing X-ray findings with metabolic data and histological features. Besides the usual features of subperiosteal resorption, diffuse demineralization and localized destruction, the less common features of osteosclerosis, and the infrequently described response of the growth plate were also demonstrated. The bone changes in roentgenographs were graded on a five point scale, 0-IV. All cases in Grade IV had parathyroid cancers and extremely high serum and urinary calcium values.

Increased excretion of urinary glycosaminoglycans in a case of Rothmund Thomson syndrome.

The exercretion of urinary total glycosaminoglycans (GAG) in a case of Rothmund Thomson syndrome associated with osteosarcoma was increased about 2--3 times that of normal control. Since the excretion of urinary total GAG in two cases of osteosarcoma was within normal range, the increased excretion of total GAG might be on account of the metabolic disorder of GAG in Rothmund Thomson syndrome. The prominent fractions obtained by Dowex 1 column chromatography from this syndrome were 0.75 M Fr and 1.0 M Fr, in which the major GAG were indicated to be partially degraded forms of heparan sulfate and chondroitin sulfate, respectively.

Hyaluronuria in a case of progeria. (Hutchinson-Gilford syndrome).

A classic case of progeria (Hutchinson-Gilford syndrome) in a 9-year-old Japanese boy is presented. The characteristic clinical features in this patient were similar to those reported in the literature. The total amount of acid glycosaminoglycans excreted in the urine was within the normal range, but there was an increase in hyaluronic acid excretion. The hyaluronuria was a novel finding in progeria, providing a common linkage with the hyaluronuria found in Werner's syndrome.

Postmortem study of a case of Werner's syndrome.

The postmortem study of a classic case of Werner's syndrome is presented. The subject was a Japanese man who died at the age of 39. Major findings included general microsplanchnia, extreme atrophy of the testes and skin, calcified aortic atherosclerosis, an increase of basophils in the pituitary, aspiration pneumonia, chronic pyelonephritis and a meningioma in the occipital area of the brain. Histologically, no osteoporosis was evident in the lumbar spine and iliac bone. The findings suggest that in Werner's syndrome the dominant pathologic factor may be found in connective tissue other than bone.