Involvement of autoimmunity to REG, a regeneration factor, in patients with primary Sjögren's syndrome.

The regenerating gene (Reg) was isolated originally as a gene specifically over-expressed in regenerating pancreatic islets and constitute a growth factor family. Reg gene product (Reg) is important in the pathophysiology of various human inflammatory diseases. Recently, the possible involvement of human REG in the regeneration of salivary ductal epithelial cells of patients with primary Sjögren's syndrome (SS) was reported. However, the expression of the REG family genes in minor salivary glands (MSG) and the occurrence of anti-REG Iα autoantibodies in SS patients were obscured. In this study, we examined the expression of REG family genes in the MSG of SS and screened anti-REG Iα autoantibodies in SS. The mRNA levels of REG family genes in MSG were quantified using real-time reverse transcription-polymerase chain reaction (RT-PCR) and REG Iα expression in the MSG was analysed by immunohistochemistry. The mRNA level of REG Iα in the MSG of SS patients was significantly higher than that of control. REG Iα protein was expressed highly in SS ductal epithelial cells. Anti-REG Iα autoantibodies in the sera were found in 11% of SS. All the MSG in the anti-REG Iα autoantibody-positive group showed REG Iα expression, whereas only 40% showed REG Iα expression in the anti-REG Iα autoantibody-negative group. The anti-REG Iα autoantibody-positive group showed significantly lower saliva secretion and a higher ratio of grade 4 (by Rubin-Holt) in sialography. These data suggest strongly that autoimmunity to REG Iα might play a role in the degeneration of MSG ductal epithelial cells in primary SS.

GTP-dependent modification of a 21-kDa substrate with NAD+ in bovine brain soluble fraction is not ADP-ribosylation of small G-protein but tailing of tRNA.

Labeling of 21-kDa material was observed when bovine brain soluble fraction was incubated with [adenylate-32P]NAD+ in the presence of GTP. The 21-kDa substrate, slightly smaller than C3 substrate in size, was labeled even without C3 exoenzyme. GTP could be replaced by nucleoside triphosphates other than ATP while ATP inhibited the GTP-induced labeling of 21-kDa substrate. After incubation of the soluble fraction with [adenylate-32P]NAD+ in the presence of GTP, [32P]ADP and [32P]ATP were detected in addition to [32P]AMP and [32P]ADP-ribose while only the last two nucleotides were observed without GTP. The 21-kDa substrate was labeled with [alpha-32P]ATP even in the absence of GTP, suggesting adenylylation rather than ADP-ribosylation. The labeled 21-kDa substrate, was extractable by phenol, disappeared with RNase treatment but not with tryptic digestion. Alkaline treatment of the phenol extract yielded an equal mixture of 3'-[32P]CMP and 2'-[32P]CMP. From these results we concluded that the 21-kDa labeling is a result of tRNA tailing with [alpha-32P]ATP generated from the [32P]AMP moiety of [adenylate-32P]NAD+. Results from reconstitution experiments using enzymes and tRNA purified from bovine brain soluble fraction, which are involved in this pathway, confirmed our conclusion.

Schizophrenia-associated idiopathic unconjugated hyperbilirubinemia (Gilbert's syndrome).

BACKGROUND: Idiopathic unconjugated hyperbilirubinemia (Gilbert's syndrome) is a benign hyperbilirubinemia found in the general population. There has been only 1 previous report of Gilbert's syndrome occurring in schizophrenic patients. The present study was conducted to determine the frequency of Gilbert's syndrome in schizophrenic patients relative to patients with other psychiatric disorders. METHOD: Plasma bilirubin concentrations of every patient admitted to the psychiatric hospital during a 3-year period were collected, and patients were examined to exclude all other causes of hyperbilirubinemia. In addition, the psychiatric symptoms of schizophrenic patients (ICD-10 criteria) with hyperbilirubinemia were evaluated by the Positive and Negative Syndrome Scale (PANSS). RESULTS: Schizophrenic patients showed a significantly higher incidence of hyperbilirubinemia (p < .05) relative to patients suffering from other psychiatric disorders, and schizophrenic patients with hyperbilirubinemia showed significantly higher scores on the positive and general psychiatric subscales of the PANSS (p < .0001) than patients without hyperbilirubinemia. CONCLUSION: The apparently higher frequency of Gilbert's syndrome in schizophrenic patients may reflect a relationship between hyperbilirubinemia and schizophrenic psychosis. Hypothetical explanations, such as a possible genetic disposition for Gilbert's syndrome, an increased vulnerability of red cell membranes, and the role of estrogens in schizophrenic patients, are discussed.

Activation of Clostridium botulinum C3 exoenzyme-catalyzed ADP-ribosylation of RhoA by K+ in a Mg2+ -dependent manner.

The effect of KCl on ADP-ribosylation of the recombinant RhoA protein catalyzed by the Clostridium botulinum C3 enzyme was studied. When the recombinant glutathione S-transferase-RhoA fusion protein (GST-RhoA) was incubated with C3 and [adenylate-32P]NAD, incorporation of radioactivity into the recombinant RhoA increased in the presence of KCl. The increase in ADP-ribose incorporation into RhoA due to KCl appeared in the presence of MgCl2 and was abolished by EDTA. C3 was stabilized by KCl, but the stabilization was also seen with BSA. The KCl-induced increase in the ADP-ribosylation was observed even in the presence of BSA during the modification reaction, thus the effect of KCl was not due to the stabilization of C3. While the initial rate of the reaction was increased by KCl, maximum incorporation of ADP-ribose per GST-RhoA molecule did not increase in the presence of KCl. Kinetic analysis revealed that KCl increased Vmax but did not alter Km for either NAD or RhoA. The NAD glycohydrolase activity of C3 was also increased by KCl. These results indicate that KCl directly activates the C3 enzyme.

Increased expression of Wnt-1 in schizophrenic brains.

The regulated expression of Wnt-1, one member of the wingless/Wnt pathway, in the brain is critical for many neurodevelopmental processes. Recently, it has been reported that the wingless/Wnt pathway participates in a complex behavioral phenomenon and suggested that this pathway's molecules are candidate genes for neuropsychiatric disorders. Thus, we investigated the expression of Wnt-1 in the hippocampal region, which is believed to be closely involved in the pathophysiology of schizophrenia, of postmortem brains from 10 schizophrenic and 10 control individuals. Immunohistochemical analysis with polyclonal antibodies recognizing Wnt-1 revealed immunoreactivity primarily in the pyramidal cell layer, particularly in CA3 and CA4 regions. We observed a significant elevation in the number of Wnt-1-immunoreactive neurons in the great majority of schizophrenic brains relative to that in controls. The expression of Wnt-1 may be related to cell adhesion, synaptic rearrangement, and plasticity. Therefore, the increase in Wnt-1 immunoreactivity in schizophrenic hippocampi suggests an altered plasticity of this structure in a large proportion of schizophrenic brains. These findings suggest an abnormality of the wingless/Wnt pathway present in the schizophrenic brain and may support the 'neurodevelopmental hypothesis' of schizophrenia.

Immunohistochemical localization of ADP-ribosylarginine hydrolase in rodent CNS.

Polyclonal antibodies were generated against ADP-ribosylarginine hydrolase (AAH), using recombinant fusion protein of rat AAH and glutathione-S-transferase as a immunogen, and affinity-purified. Western blotting showed that the antibodies recognized in mouse brain homogenate a single protein with a molecular mass of 38 kDa, the expected size for mouse AAH. An analysis using the antibodies revealed that heavy labelings were apparent in various brain regions. In the cerebral cortex, pyramidal cells in layers III and V were the most heavily labeled. In the hippocampal formation, labeling was present on the pyramidal neurons and granule cells. The most heavily immunostained cell type was the pyramidal neuron of CA3. In the cerebellum, Purkinje cells were the most heavily labeled. Less intense staining was present over the granule cells. In the basal ganglia, neurons in the caudate nucleus and large multipolar cells in the amygdaloid complex were immunoreactive. Heavy labeling was seen in many midbrain and brainstem nuclei. Neurons in the habenula and ependymal cells were stained heavily. On Western blot analysis of rat cerebrospinal fluid (CSF), the anti-AAH antibodies recognized a protein with a molecular mass of 38 kDa. This is apparently the first evidence of a widespread but distinctive distribution of AAH in neurons of mouse brain and the presence of extracellular AAH in rat CSF.

Region-specific changes of GABAA receptors by tolerance to and dependence upon pentobarbital.

An experimental model for inducing tolerance to and dependence upon pentobarbital was characterized. Rats were infused with pentobarbital, 300 micrograms/10 microliter per h i.c.v. for 6 or 7 days, through pre-implanted cannulas by osmotic minipumps. Measurement of brain and serum levels showed that pentobarbital remained mainly inside of the brain. Measurements of sleeping time and susceptibility to convulsant-induced seizures indicated a substantial degree of tolerance and dependence (24 h after termination of infusion). Results of GABAA receptor binding assays showed marked regional variations. While [35S]t-butylbicyclophosphorothionate (TBPS) binding sites were increased in frontal cortices and striata of pentobarbital-dependent animals, KD was increased in striata of tolerant animals. Dependence upon pentobarbital was correlated with increased [3H]flunitrazepam binding sites in all three regions examined. Both high and low affinity [3H]muscimol binding sites were increased in dependent animals, but low affinity sites were decreased in frontal cortices of tolerant animals. KDs of [3H]muscimol high affinity sites were increased in cerebellum after animals developed dependence upon pentobarbital. KDs of [3H]muscimol low affinity sites were decreased in striata of pentobarbital tolerant animals. These findings further support the hypothesis that GABAA receptors in discrete areas of the brain have different subunit compositions and are regulated differentially by pharmacological modulators.

Parkinson's disease associated with argyrophilic grains clinically resembling progressive supranuclear palsy: an autopsy case.

A 70-year-old male began to show akinesia, rigidity of extremities, finger tremor, disturbed vertical external ocular movement, and nuchal dystonia, which progressed slowly. Brain CT scan and magnetic resonance images showed slight atrophy of the frontal lobe and slight enlargement of the lateral ventricles. Hasegawa's dementia rating scale-revised version gave a moderate score of 11/30 points. He died of pneumonia at the age of 76. The clinical diagnosis was progressive supranuclear palsy (PSP). However, there were no neuropathological characteristics of PSP. Neuropathologically, Parkinson's disease was diagnosed. In addition, many argyrophilic grains (ArGs) in the gray matter were stained, especially in the insula, amygdala, hippocampus, parahippocampal gyrus, lateral occipitotemporal gyrus, and substantia nigra, by the Gallyas-Braak method. We consider that ArGs could modify the symptoms of Parkinson's disease and that Parkinson's disease with ArGs may show a PSP-like clinical course.

Effects of pentobarbital tolerance and dependence on convulsant and GABAA receptor antagonist binding.

Experiments were performed which examined the effects of pentobarbital tolerance and dependence on GABAA receptor antagonist binding. In rats implanted with pentobarbital pellets for 7 days, followed by 24 hours of withdrawal, there was a significant decrease in the latency of TBPS-induced seizures and an increase in [35S]TBPS binding in the frontal cortex. The pentobarbital tolerant rats had a significant increase in the low affinity KD of [3H]SR95531 binding. Removal of the pellets for 24 hours caused a reversal of the effect on the low affinity KD and caused a decrease in the number of low affinity binding sites. In vitro addition of pentobarbital to binding assays produced a decrease in the number of high affinity [3H]SR95531 binding sites without changing low affinity binding. In the cerebellum, the binding in none of the treatment groups was significantly different from placebo. These observations suggest that pentobarbital tolerance and withdrawal cause changes in the properties of the GABAA receptor antagonist binding site which are different from those caused by in vitro exposure to the drug.

Reversible pitch perception deficit caused by carbamazepine.

Carbamazepine (CBZ) is an antiepileptic drug frequently used to treat a variety of neurologic diseases or symptoms. In addition, the drug is used as a mood stabilizer in patients with affective or schizophrenic disorders. Among its adverse effects, auditory disturbance is described rarely. In this report, we describe a 25-year-old woman who noted falsely higher pitch perception after starting CBZ treatment for schizoaffective disorder. We also review the literature reporting CBZ-associated abnormal pitch perception.

Contribution of sodium valproate to the syndrome of inappropriate secretion of antidiuretic hormone.

We report the case of a 62-year-old man who was administered sodium valproate (VPA) and who subsequently developed the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). He had been taking VPA for treatment of idiopathic generalized tonic-clonic convulsions since he was 56 years old. After substituting VPA with zonisamide, the serum sodium level returned to normal. We consider this episode of SIADH to be the result of a combination of factors including a weakness of the central nervous system and the long-term administration of VPA.

Two-year oral chronic toxicity and carcinogenicity study in rats of diets fumigated with methyl bromide.

A chronic toxicity and carcinogenicity study was conducted by feeding diets containing 80, 200 or 500 ppm total bromine following fumigation with methyl bromide, a diet containing 500 ppm potassium bromide, or a basal diet alone to groups of 60 male and 60 female Fischer (F344) rats for up to 2 yr. Ten males and ten females from each group were killed after wk 52 and 104 for urinalysis, haematology, blood biochemistry and pathology. Rats that were killed in extremis or found dead during the study and all rats that survived to the end of the study were also subjected to pathological examinations. In rats fed the diets fumigated with methyl bromide there were no marked toxic changes, except for a slight depression of body-weight gain from wk 60 onwards in males of the 500-ppm group, and tumour incidence was unaffected. Rats given a diet containing potassium bromide did not show any treatment-related changes. It was concluded that residues of up to 500 ppm total bromine in diets fumigated with methyl bromide are not carcinogenic in F344 rats of either sex and that the maximum no-effect level is 200 ppm (6.77 mg total bromine/kg body weight/day) in males. The maximum no-effect level could not be determined in females.

Thirteen-week subacute toxicity study of 4-bromophenyl chloromethyl sulfone (BCS) in rats.

Four groups of 12 Wistar SPF rats of both sexes were fed diets containing 4-bromophenyl chloromethyl sulfone (BCS) at the levels of 0, 20, 100 or 500 ppm for 13 weeks to evaluate the subacute toxicity. Males and females in the 500 ppm group showed increases in plasma protein and calcium and a decreased A/G ratio. The values of plasma total cholesterol and uric acid increased in this female group. In addition, significant increases in both absolute and relative weights were seen in the liver, thyroid and adrenal of this group of both sexes, together with the increased kidney weight in males. All males and females in the 500 ppm group had centrilobular hepatocellular swelling of the liver consisting of proliferation of the smooth endoplasmic reticulum. Males of this group also showed increased incidences of hyalin droplet degeneration of the proximal tubular epithelium and focal tubular atrophy of the kidney. The maximum no-effect level of BCS was determined to be 100 ppm (males: 6.25 mg/kg/day, females: 6.80 mg/kg/day).

Quantitative comparison between serum components and somatic cells in bovine quarter milk.

Quantitative comparisons between serum components and somatic cells in 202 bovine quarter milks collected at 3 dairy farms were conducted. Detection of serum components was carried out by an immunological technique, and cell counts were calculated by Breed's method. A high correlation (r = 0.77) between cell counts and gamma-globulin contents was demonstrated, while correlation between cell counts and serum albumin was not close (r = 0.45). In a comparison between cell counts and gamma-globulin contents, 19 (65.4%) out of 29 milks containing over 500,000 cells/ml had gamma-globulin less than 1.138 mg/ml, which was also seen in 97% of 172 milks having less than 500,000 cells/ml. From these results, it was suggested that the gamma-globulin content of milk might be of use for differentiating normal milk from abnormal ones.

[A case of a tumor of the spermatic cord metastatic from cecal cancer].

A 71-year-old man was admitted with the complaint of painless tumor in the right inguinal region, one month after right hemicolectomy for cecal cancer. The tumor seemed to exist in the right spermatic cord, so right radical orchiectomy was done. A tumor was found in the right spermatic cord, but the right spermatic duct, right epididymis and right testicle were intact. Histopathological examination of the tumor revealed metastatic adenocarcinoma from the cecal cancer. Twenty cases of metastatic tumors of the spermatic cord from the gastrointestinal cancers have been reported in Japan including this case.

[Effects of muscle relaxant E.M.P.P. on afferent discharges of muscle spindle in man--an microneurographic analysis (author's transl)].

Effects of a new muscle relaxant E.M.P.P.(4'-ethyl-2-methyl-3-piperidino-propiophenone . hydrochloride) on afferent discharges of muscle spindle were studied in 10 healthy volunteer subjects. Single unit afferent discharges of muscle spindle were recorded microneurographically from the median or tibial nerve using a tungsten microelectrode with a tip diameter of about 1 micrometer (impedence: about 10 M omega). Effects of an oral administration of 150--300 mg of E.M.P.P. on spontaneous afferent discharges as well as dynamic and static responses of muscle spindle were analysed in the present study. 1. E.M.P.P. suppressed remarkably the frequency of spontaneous afferent discharges of muscle spindle. Meanwhile, mental activations such as speech of mental arithmetic recovered temporarily the frequency of afferent discharges to that before E.M.P.P. administration. 2. E.M.P.P. suppressed the dynamic and static responses of muscle spindle to stretch. It also suppressed the dynamic responses of muscle spindle in the decontraction phase of electrically-induced twitch contractions of the receptor-bearing muscle. Based on these results, it is concluded that E.M.P.P. suppresses the static and dynamic activities of muscle spindle in man. These effects seem to be due to modifications of descending influences from central structures on the static and dynamic gamma motoneurons which innervate the muscle spindle.

[Equal-sensation contours and a threshold curve for low-frequency vibrotactile stimuli on the glabrous skin of the human hand].

Using vibrotactile stimuli, equal-sensation contours and a threshold curve were determined on the glabrous skin of the human hand. Equal-sensation contours were measured between 1 Hz and 16 Hz. When plotted on log-log coordinates, they monotonically decreased as a function of frequency and were downward by concaved. The threshold curve was measured between 1 Hz and 50 Hz. Its values decreased linearly from 1 Hz to 16 Hz, became constant at 16-35 Hz, and decreased again at 35-50 Hz. Therefore, at the stimulus range of 1-16 Hz, equal-sensation contours were downward concave and the threshold curve was more or less linear. This discrepancy seems to be caused by the differences in mechanoreceptor types which take part in subjective responses: suprathreshold sensation depends on several types of mechanoreceptors, whereas vibrotactile thresholds depends mainly on RA receptors (Meissner corpuscles).

Characteristic brain hypoperfusion by 99mTc-ECD single photon emission computed tomography (SPECT) in patients with the first-episode schizophrenia.

OBJECTIVE: In this study, we evaluated brain perfusion in patients with first-episode medicated schizophrenia using the new analytical method, statistical parametric mapping (SPM) applied to single photon emission computed tomography (SPECT). METHOD: We performed SPECT with 99-Tc-ethyl cysteinate dimer (99mTc-ECD) of the brain and magnetic resonance imaging (MRI) in patients with schizophrenia (n=30) and control subjects matched for age and gender (n=37). A voxel-by-voxel group analysis was performed using SPM2 (Z>3.0, P<0.001, uncorrected for multiple comparisons). RESULT: In comparison with control subjects, the volumes of the bilateral frontal areas were found to be decreased on MRI. Blood flow was found to be reduced in the bilateral temporal areas in the patients with schizophrenia on SPECT. CONCLUSION: In this study, patients with first-episode schizophrenia appeared to have significant bilateral temporal hypoperfusion, although temporal volumes were not significantly decreased in comparison with control subjects. Abnormality of temporal lobe blood flow in schizophrenia may show that functional changes occur earlier than structural changes, and may assist in the diagnosis of schizophrenia.