Expression of a novel matrix metalloproteinase gene during Cynops early embryogenesis.

Matrix metalloproteinases (MMPs) are thought to play important roles in the gastrulation of Cynops pyrrhogaster embryos. MMP cDNAs were cloned from Cynops pyrrhogaster and we report here a novel MMP called CyMMP, which has strong similarity to MMP-21 (XMMP) in Xenopus. Reverse transcription-polymerase chain reaction (RT-PCR) analysis demonstrated that CyMMP mRNA was already present in cleavage stage embryos. The amount of the mRNA then gradually decreased, but increased again starting in late gastrula. There were regional differences in the level of CyMMP mRNA expression at late gastrula: the involved archenteron roof was the predominant site of expression of the gene, while there was weak expression in the neuroectoderm and epidermal ectoderm. We also found that the gene was activated in artificially mesodermalized ectoderm. The present findings indicate that CyMMP mRNA expression is activated in differentiating mesoderm during gastrulation, suggesting that CyMMP plays a role in gastrulation-related cell movement.

Mutational analysis of the anion exchanger 3 gene in familial paroxysmal dystonic choreoathetosis linked to chromosome 2q.

Familial paroxysmal dystonic choreoathetosis (PDC) is an autosomal dominant neurological disorder characterized by episodes of involuntary movement precipitated by caffeine, alcohol, or emotional stress. The locus for PDC has recently been mapped to chromosome 2q32-36, but its causative gene has not yet been identified. PDC is most likely a kind of channelopathy, as suggested by the fact that other paroxysmal neurological disorders are caused by various ion channel mutations. Although no ion channel is located in this candidate region, anion exchanger 3 (AE3) has been mapped to 2q36 and has also been reported to be the most promising candidate gene of PDC. In this study we performed sequencing of the coding region of the AE3 gene in patients with familial PDC linked to chromosome 2q and excluded the AE3 gene as the causative gene for PDC. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 88:733-737, 1999.

Familial paroxysmal dystonic choreoathetosis: clinical findings in a large Japanese family and genetic linkage to 2q.

BACKGROUND: Paroxysmal dystonic choreoathetosis (PDC) is a rare familial movement disorder that has been mapped to chromosome 2q31-36. OBJECTIVE: To study the first Japanese family with PDC clinically and genetically. PATIENTS AND METHODS: We studied a large Japanese family in which at least 17 members in 6 generations have been affected by PDC. We interviewed and examined 26 family members, 8 of whom revealed choreoathetosis-like and dystonialike involuntary movement and 1 of whom revealed no involuntary movement but only muscle stiffness such as the aura of paroxysmal dystonic choreoathetosis (PDC). Genetic linkage studies of this family were carried out with polymorphic DNA markers. RESULTS: The attacks of involuntary movement or muscle stiffness were precipitated by ovulation, menstruation, emotional stress, or caffeine or alcohol ingestion. Magnetic resonance imaging of the brain revealed no abnormalities. Clonazepam therapy was effective for reducing the attacks, and ingestion of garlic was believed by patients to be effective for softening the attacks. An affected woman with only muscle stiffness showed remission after hysterectomy for hysteromyoma. This woman also had the disease haplotype and transferred it to her typical PDC-affected daughter. Maximal pairwise logarithm of odds scores exceeding 2.00 were obtained at D2S2250, D2S1242, D2S377, D2S2148, and D2S126. The PDC gene was demonstrated by linkage analyses to be located in a 15.3-centimorgan interval lying between D2S371 and D2S339 based on pairwise and multipoint logarithm of odds scores and obligate recombination events in affected individuals. CONCLUSIONS: Linkage of PDC to chromosome 2q32-36 was confirmed in a Japanese family. The clinical characterizations of this family with PDC include that ovulation seems also to be a precipitating factor of the attacks and that hysterectomy seems to be effective for softening the attacks. Although low-dose clonazepam treatment was most effective, garlic use was believed by affected members to be effective for softening the attacks. Furthermore, based on the results of clinical and genetic analyses, we suggest that muscle stiffness without involuntary movement may represent a forme fruste of PDC.

Three cases of spinal decompression sickness treated by U.S. Navy Treatment Table 7.

For patients of type 2 decompression sickness, recompression therapy using U.S. Navy Treatment Table 6 (TT6) and its extensions is the most common means of treatment. However, some cases are resistant to the recompression therapy, and the outcome of TT6 is not always satisfactory. Although a new table, the U.S. Navy Treatment Table 7 (TT7) was described in 1985 in the U.S. Navy Diving Manual, to date few cases who were treated using TT7 have been reported. Here, we report three cases of spinal decompression sickness who received treatment according to TT7. Two were sports scuba divers, and the other a commercial diver. TT7 was applied later than 4 d after onset in all three cases; two patients were remarkably improved during the recompression therapy, while the other improved to a certain extent after additional repetitive TT6. Mild impairment of lung function, probably due to pulmonary oxygen toxicity, was observed on lung function testing in one case. In all cases, after additional TT6 and/or rehabilitation, patients were able to return to active daily living.

Emerin, deficiency of which causes Emery-Dreifuss muscular dystrophy, is localized at the inner nuclear membrane.

X-linked recessive Emery-Dreifuss muscular dystrophy (EDMD) is an inherited muscle disorder characterized by the clinical triad of progressive wasting of humero-peroneal muscles, early contractures of the elbows, Achilles tendons and postcervical muscles, and cardiac conduction block with a high risk of sudden death. The gene for EDMD on Xq28 encodes a novel protein named emerin that localizes at the nuclear membrane of skeletal, cardiac and smooth muscles and some other non-muscle tissues. To investigate a possible physiological role for emerin, we examined the ultrastructural localization of the protein in human skeletal muscle and HeLa cells, using ultrathin cryosections. We found that the immune-labeled colloidal gold particles were localized on the nucleoplasmic surface of the inner nuclear membrane, but not on the nuclear pore. Emerin stayed on the cytoplasmic surface of the nuclear lamina, even after detergent treatment that solubilizes membrane lipids and washes out membrane proteins. These results suggest that emerin anchors at the inner nuclear membrane through the hydrophobic stretch, and protrudes from the hydrophilic region to the nucleoplasm where it interacts with the nuclear lamina. We speculate that emerin contributes to maintain the nuclear structure and stability, as well as nuclear functions, particularly in muscle tissues that have severe stress with rigorous contraction-relaxation movements and calcium flux.

[A Japanese family with paroxysmal dystonic choreoathetosis].

We present the first report of Japanese family with paroxysmal dystonic choreoathetosis (PDC). At least seventeen individuals of this family in six generations were affected by PDC by autosomal dominant inheritance. The affected individuals had attacks of choreoathetosis/dystonia-like involuntary movements without loss of consciousness. These attacks were precipitated by stress, caffeine, menstruation or ovulation, but not by sudden voluntary movements, which are common precipitating factors in paroxysmal kinesigenic choreoathetosis (PKC). MRI studies of the brain revealed no abnormalities. Some of the family members received clonazepam, or garlic with significant therapeutic effects, while one female case improved after hysterectomy. The pathogenesis of PDC remains unknown, but it could be attributed to a form of channelopathy. Recently, two linkage studies were reported and the PDC locus was identified on chromosome 2q, which may lead to clarify the PDC gene. The family described here may be important for the future biochemical and genetic analyses. We also suspect that PDC may relate to endocrinological abnormalities besides channelopathy.

[A case of vertical gaze palsy associated with a unilateral infarct in the thalamo-mesencephalic junction on MR imaging].

We report a case of a 54-year-old man with sudden-onset double-vision. On admission, neurological examination showed upward and downward gaze palsy on voluntary and smooth pursuit movements, and vertical oculocephalic maneuver elicited a full upward and downward response. Bell's phenomenon, horizontal eye movements and convergence were not impaired. Based on these findings, supranuclear dissociated vertical gaze palsy was diagnosed. T1-weighted MR images revealed low intensity on the medial side of the right thalamo-mesencephalic junction, which impaired the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF). T2-weighted MR images revealed a high-intensity area. The posterior commissure was spared. The clinical signs gradually improved, and the vertical gaze palsy almost disappeared one month after onset. Based on these findings, unilateral infarct in the thalamo-mesencephalic junction in the distribution of the right paramedian thalamic artery was diagnosed. Only two cases of upward and downward gaze palsy in association with unilateral upper midbrain lesion without posterior commissure have been previously reported. Since we did not perform a pathological examination, we cannot deny that there may have been some denervation of fibers at the posterior commissure. Cases of upward and downward gaze palsy in association with unilateral upper midbrain lesion without posterior commissure are rare, and it is very interesting that the lesion in our patients, like that seen in the two pathological reports, was right-sided.

Dystrophin-related protein is found in the central nervous system of mice at various developmental stages, especially at the postsynaptic membrane.

Dystrophin deficiency is known to be the cause of X-linked Duchenne muscular dystrophy (DMD). A recently cloned B3-cDNA shares 80% homology with the C-terminus and actin-binding portion of dystrophin. This autosome-derived gene product is called dystrophin-related protein (DRP). DRP is known to exist in fetal muscles even in mdx mice, an animal model for X-linked DMD, but not in mature mouse muscles. We raised a polyclonal antibody against a B3-unique amino acid sequence (Ab-LDP) and investigated the existence and distribution of DRP in the central nervous system (CNS) tissues of mdx and normal control B10 mice at various stages of development using immunoblotting and immunohistochemical methods. The former shows that DRP exists in the CNS of both B10 and mdx mice, regardless of the developmental stage, with the exception that the 420 kDa DRP band of the 15-day fetus is faint. In immunohistochemical studies, the choroid plexus, some neurons, glial cels, pia mater, and blood vessels were stained with Ab-LDP. Staining intensity did not differ between B10 and mdx mice or between developmental stages except that the 15-day fetus stained only faintly. This is in contrast to the results obtained for muscles in which DRP localized to muscle membrane in embryo decreases and is assembled at the neuromuscular junction in adults. In addition, an electron microscopic study on the cerebral cortex from adult B10 mice was also performed and revealed Ab-LDP staining of the postsynaptic membrane of dendrite and the rough endoplasmic reticulum of neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

MR imaging findings of tremors associated with lesions in cerebellar outflow tracts: report of two cases.

Two cases with severe tremors were studied by means of electromyograms using surface electrodes and also by magnetic resonance (MR) imaging. The first case was associated with multiple sclerosis and demonstrated a severe postural cerebellar tremor and an alternate activation of antagonist muscles in the right arm. The second case, with hemorrhage in the brainstem, demonstrated a severe tremor at rest and mixed synchronous and alternating activation of antagonist muscles in the left forearm. MR imaging studies localized lesions possibly responsible for these tremors. In the first case a lesion was located in the superior cerebellar peduncle just under the decussation, and in the second case a lesion was found between the red nucleus and the thalamus, with possible involvement of both the cerebellothalamic and nigrostriatal pathways. The first case accords with the theory that a lesion located in the dentate nucleus and its projection can cause severe postural cerebellar tremor. The lesion demonstrated in the second case may be responsible for "cerebellar tremor at rest" or "static (resting) cerebellar tremor."

[A case of acute myocardial infarction due to coronary spasm].

The patient was a 47 year-old man, who has been known to have effort angina since September 1989. His exercise stress ECG has revealed ST elevation in V2-V4 with maximum exercise. He experienced severe chest pain lasting for an hour on the way to his office in the early morning on November 16, 1989, and was admitted to our hospital. His ECG and laboratory findings indicated typical acute anteroseptal myocardial infarction, but the coronary arteriography (CAG) which was performed 7 hours after the onset showed no significant stenotic lesion. After administrating nitrate and calcium antagonist, he has had no attack of angina pectoris and his exercise stress test has revealed no ST-T changes on his ECG. 1 month later, while antianginal drugs were discontinued in order to perform an ergonovine stress test, the patient frequently complained of left anterior chest pain with remarkable ST elevation in precordial leads on his ECG. The CAG at chronic stage revealed that there was a 99% stenosis at Segment 6 of the left anterior descending artery (LAD) which was supplied with good collateral flow from the right coronary artery. The LAD was completely occluded at Segment 6 after intracoronary administration of ergonovine maleate 0.005 mg to the left coronary artery. After the intracoronary infusion of isosorbide dinitrate, there was no significant stenosis seen in the LAD except the minimum wall irregularity at Segment 6. These findings suggested that coronary spasm might play a major role of the occurrence of acute myocardial infarction in this case.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrastructural features of acidic complex carbohydrates in the intercellular matrix of the ovarian follicles in adult mice.

In the intercellular matrix of the granulosa layer of the mouse ovarian follicles, ultrastructural features of acidic complex carbohydrates have been studied by means of dialyzed iron (DI) staining in combination with procedures of digestion with Streptomyces and testicular hyaluronidases. In the intercellular matrix, DI reactive structures containing acidic complex carbohydrates consist of layers of a variable thickness coating the plasma membrane of the granulosa cells and reticular elements distributed in the spaces between the cells. The latter exists in two appearances; one is clumped masses of irregular shapes and different sizes, whereas the other being filamentous figures radiating from the masses. The effects of digestion with Streptomyces and testicular hyaluronidases upon the DI staining of the tissues indicate that the DI reactive structures in the intercellular matrix contain at least three types of acidic complex carbohydrates; hyaluronic acid, isomeric chondroitin sulfates and other acidic glycosaminoglycans. The histophysiological activities played by these particular complex carbohydrates have been briefly discussed.

The histochemistry of complex carbohydrates in the ovarian follicles of adult mice.

In the ovarian follicles of adult mice, complex carbohydrate-containing structures have been studied by means of light microscopic histochemical methods. In the ovarian follicles, the zona pellucida of oocytes, follicular fluid and intercellular matrix of the granulosa layer are found to exhibit positive reactions for complex carbohydrates with 1,2-glycol and acidic groups and alpha-D-mannosyl and alpha-D-glucosyl residues. The present histochemical analyses have revealed that the complex carbohydrates common to the three types of the structures are hyaluronic acid, sulfated glycosaminoglycans other than isomeric chondroitin sulfates and neutral glycoproteins and that sialic acid is a particular moiety of the zona pellucida, whereas isomeric chondroitin sulfates being that of the follicular fluid and intercellular matrix of the granulosa layer. The histophysiological activities of the carbohydrate-containing structures have been discussed with special reference to their histochemical properties determined in the present study.