Zimmermann-Laband syndrome: a case report.

Zimmermann-Laband syndrome is a very rare disorder characterized by gingival fibromatosis, abnormalities of soft cartilages of the nose and/or ears, hypoplastic or absent nails and terminal phalanges, joint hypermobility, hypatoslenomegaly, mild hirsutism and learning difficulties. Early presentation of Zimmermann-Laband syndrome in a newborn has rarely been described. This paper describes a newborn patient with Zimmermann-Laband syndrome.

Expression of osteoclast differentiation factor and osteoclastogenesis inhibitory factor in rat osteoporosis induced by immunosuppressant FK506.

Immunosuppressant drugs are currently required by transplant recipients for the remainder of their lives, despite the many adverse effects associated with these therapies. Acute osteoporosis is one such effect, and a reproducible osteoporosis model has been established through the administration of the immunosuppressant drug FK506 in rats. The cause of this osteoporosis has been shown to be abnormal osteoclast proliferation, altering the process of bone remodeling. However, the reasons why FK506 induces osteoclast proliferation and whether this process is mediated by cytokine changes or an increase in bone resorption factors have been unclear. An investigation was therefore conducted focusing on the recent discoveries of osteoclast differentiation factor (ODF) and osteoclastogenesis inhibitory factor (OCIF). These factors led to elucidation of the osteoclast differentiation-maturation mechanism. An osteoporosis model was produced in rats utilizing intramuscular FK506 injection (1 mg/kg) for 28 consecutive days. Trabecular bone resorption was observed inferior to enchondral ossification in the FK506 group, and tartrate resistant acid phosphatase (TRAP) staining revealed a clear increase in osteoclasts at the site of enchondral ossification, relative to the control group. Real-time PCR and in situ hybridization (ISH) demonstrated minimal differences in OCIF expression between control and the treatment groups. However, Real-time PCR revealed clearly increased ODF expression in the treatment group. ODF expression was also shown to be increased in the treatment group using ISH. This was histologically consistent with a region of osteoclast proliferation inferior to enchondral ossification. The results of this study support the hypothesis that FK506-mediated osteoporosis occurs by action of the drug on osteoclasts, promoting expression of ODF messenger ribonucleic acid (mRNA) and thus prompting osteoclast differentiation and maturation.

Immunohistochemical study on expression of alpha-defensin and beta-defensin-2 in human buccal epithelia with candidiasis.

OBJECTIVES AND DESIGN: It has been previously reported that alpha-defensin (HNPs) and beta-defensin-2 (HBD-2) peptides with antifungal and cytotoxic activities can be detected in oral carcinomas and the saliva of patients with oral carcinomas. The present study investigated the presence of HNPs and HBD-2 in oral epithelia with candidiasis. MATERIALS AND METHODS: Tissue sections (4 microm) were prepared from biopsy and surgically removed specimens diagnosed as oral candidiasis (n = 10). The sections were examined immunohistochemically with antibodies directed against HNPs and HBD-2. RESULTS: Tissue sections of oral candidiasis were immunostained with antidefensin antibodies. Neutrophils in the inflamed lamina propria were positively immunostained with anti-HNPs antibody. The cytoplasm of cells in the upper spinous layer, in the lower spinous layer and in the parakeratinized layer of buccal epithelia with candidiasis was immunostained intensely with anti-HBD-2 antibody. In contrast, the expression of HBD-2 in the normal spinous layer was much weaker than that in oral candidiasis. No signals of HNPs were found in normal buccal epithelium. CONCLUSION: Buccal specimens from individuals with oral candidiasis show greater levels of expression of both HNPs and HBD-2. There might be a dual protection manner by defensins against fungal inflammation in infected buccal epithelia locally. Generally, HBD-2 signals have been found everywhere in the buccal epithelium; however, in an infected area, the signal intensity of HBD-2 has increased. HNPs signals have not been found in the normal buccal epithelium; however, HNPs signals have increased when the infection occurred.

Human alpha-and beta-defensin immunoreactivity in oral mucoepidermoid carcinomas.

The purpose of this study was to demonstrate the immunohistochemical localization and distribution of human alpha- and beta-defensins, peptides with antimicrobial activity, in oral mucoepidermoid carcinoma tissue. Tissue samples were embedded in paraffin and alpha- and beta-defensins were immunostained by the streptavidin-biotin coupled peroxidase method. Cancer cells that constituted the ducts, as well as neutrophils, were positively immunostained with the anti-alpha-defensin antibody (HNPs). On the other hand, epidermoid cells and intermediate cells were intensely stained with the anti-beta-defensin-2 (HBD-2) antibody. Mucous-secreting cells were clearly not immunostained with the anti-HBD-2 antibody. The epithelial hyperplasia region adjacent to the tumor tissues was also positively immunostained with the anti-HBD-2 antibody.

Presence of human beta-defensin-2 in oral squamous cell carcinoma.

The purpose of this study was to demonstrate immunohistochemically the localization and distribution of human beta-defensin-2 (HBD-2), a peptide with antimicrobial activity, in oral carcinoma tissues. Tissue samples were embedded in paraffin, and HBD-2 was immunostained by the streptavidin-biotin coupled peroxidase method. Cancer cells in the cornified region of well differentiated squamous cell carcinomas were stained intensely. Stained cancer cells detected by anti-HBD-2 antibody were scattered among the cells of the non-cornified region.

Pharmacological characterization of an 18-kDa protein associated with the peripheral-type benzodiazepine receptor in salivary glands.

Pharmacological characterization of peripheral type benzodiazepine receptors in rat, rabbit, mouse and human salivary glands was determined by receptor binding and photoaffinity labeling analysis using [3H]PK14105 (1-(2-fluoro-5-nitrophenyl)-3-isoquinolinecarboxylic acid). [3H]PK14105 bound to the membranes of salivary glands in rats, rabbits, mice and humans with high affinity at the nanomolar level. The rank order of receptor density in submandibular glands among several species was as follows: human > or = rat > or = mouse > rabbit. Competitive potency of receptor ligands against [3H]PK14105 was as follows: PK1195 > or = Ro5-4864 > diazepam > clonazepam > Ro15-1788. The rank order of potency against calcium channel ligands and co-transport inhibitors was as follows: nitrendipine > BAY K 8644 > bumetanide > furosemide. Pretreatment with nitrendipine or BAY K 8644 decreased the affinity of [3H]PK14105 binding to rat parotid gland membranes, without changing the density. The photoaffinity labeling with [3H]PK14105 indicated the presence of the 18-kDa protein in all salivary glands of our experiment. The inhibition of photolabeling by some receptor ligands was the same results as the receptor binding assay. In conclusion, the peripheral type benzodiazepine receptors include the 18-kDa protein photolabeled with [3H]PK14105 in salivary glands of rat, mouse, rabbit and human.

Effects of anticonvulsants on local anaesthetic-induced neurotoxicity in rats.

The effects of various anticonvulsants on local anaesthetics procaine- and lidocaine-induced convulsions were investigated in rats. Pretreatment with diazepam (2.5-5 mg/kg, intraperitoneally) and clonazepam (5-10 mg/kg, intraperitoneally) completely protected the rats against both local anaesthetic-induced convulsions. Phenobarbital (12.5-50 mg/kg, subcutaneously) also significantly decreased the incidence of both convulsions and prolonged their latencies. Carbabazepine (10-40 mg/kg, intraperitoneally) did not completely repress both convulsions, but it prolonged their latencies. Phenytoin (5-20 mg/kg, intraperitoneally) and primidone (30-60 mg/kg, intraperitoneally) markedly enhanced both local anaesthetic-induced convulsions, as shown by shortening of latency and increase in mortality. Valproate (100-200 mg/kg, intraperitoneally) produced a protective effect against procaine-induced convulsions, while it strongly enhanced lidocaine-induced convulsions. These results suggest that the benzodiazepines are effective drugs to prevent neurotoxicity induced by local anaesthetics, while phenytoin and primidone potentiate them.

[A case of teratoma in both the mediastinum and the intrapulmonary system].

A rare case of mature teratoma in both the mediastinum and the intrapulmonary system is presented. A 30-year-old male was admitted to our hospital due to tumor masses in the mediastinum and the left lung. We performed mediastinal tumor resection and left upper partial lobectomy. Neither tumor communicated with each other. Pathological findings revealed teratoma in the mediastinal lymph node and the intrapulmonary system including no malignant cells in either tumor. In this case, because metastasis and perforation were negative, we proposed that both tumors occurred at the same time in the early embryo.

Relationship between pathogenicity of Coxiella burnetii isolates and gene sequences of the macrophage infectivity potentiator (Cbmip) and sensor-like protein (qrsA).

Coxiella burnetii, the Q fever agent, is an obligate intracellular bacterium and survival in phagolysosomes is an important virulence factor. The present study was performed to determine the relationship between its pathogenicity and genes related to its survival in macrophages, i.e. macrophage infectivity potentiator and Q fever agent regulatory sensor-like protein. The sequence similarity was more than 99% among Japanese, European and American strains, and no relationship was found between pathogenicity in guinea pigs and these genes.

Homogeneity of Borrelia japonica and heterogeneity of Borrelia afzelii and 'Borrelia tanukii' isolated in Japan, determined from ospC gene sequences.

Borrelia afzelii, B. japonica, and 'B. tanukii' isolated from various sources and geographical origins in Japan were characterized by restriction fragment length polymorphism (RFLP) analysis and sequencing analysis of the outer surface protein C (OspC) amplicon. B. afzelii and 'B. tanukii' generated variable RFLP patterns and differences in ospC gene sequence were confirmed. In contrast, 26 isolates of B. japonica generated one OspC RFLP type, and sequence similarity between B. japonica ranged from 96.4 to 99.7%. These finding suggests that B. japonica is unique in comparison with other members of B. burgdorferi sensu lato species with respect to homogeneity of the ospC gene.

Identification of rickettsiae isolated in Japan as Coxiella burnetii by 16S rRNA sequencing.

The 16S rRNA genes of Japanese Coxiella isolates obtained from various sources and geographical areas were directly sequenced by dideoxynucleotide chain termination methods in which Taq DNA polymerase was used. The levels of sequence similarity among Japanese, European, and American isolates were more than 99%, and the Japanese isolates were identified as Coxiella burnetii, C. burnetii strains isolated worldwide, including Japan, were found to be very similar.

Pharmacological analysis of local anaesthetic tolycaine-induced convulsions by modification of monoamines in rat brain.

The effects of a local anaesthetic, tolycaine, on brain monoamine levels were investigated during the convulsive process in rats. The influence of central monoamine modifications on tolycaine-induced convulsions was also examined. Tolycaine (140 mg/kg, intraperitoneally) produced a significant elevation of noradrenaline and 5-hydroxytryptamine levels in all brain regions in the convulsive state from the levels in the non-convulsive state. Their levels returned to normal during the postconvulsive state. Dopamine levels were depleted in the cerebral cortex, the striatum, and the ponsmedulla oblongata during the convulsive process and increased in the cerebellum. Pretreatment with alpha-methyl-p-tyrosine, which depletes brain catecholamine, suppresses the tolycaine-induced convulsions, as shown by a decrease in the incidence; L-3,4-dihydroxyphenylalanine and bis-(1-methyl-4-homopiperazinyl-thiocarbonyl)-disulfide, which increase brain catecholamine, intensified the convulsions, as shown by shortening of the latency and increase in the mortality. Antagonists of beta-adrenergic and dopamine receptors, such as propranolol, chlorpromazine and pimozide, markedly suppressed the convulsions, but an antagonist of alpha-adrenergic receptor, phenoxybenzamine, had no effect. Furthermore, 5-hydroxytryptophan, which increases brain 5-hydroxytryptamine, suppressed the convulsions, and DL-p-chlorophenylalanine, which depletes brain 5-hydroxytryptamine, intensified them. Antagonists of 5-hydroxytryptamine receptor, methysergide and methiothepin, suppressed the convulsions. These results suggest that brain noradrenaline and 5-hydroxytryptamine are major regulators in the tolycaine-induced convulsive process and that central catecholaminergic neurones act in a stimulatory way on the tolycaine-induced convulsions, while serotonergic neurones act suppressively.

Decreased m3-muscarinic and alpha 1-adrenergic receptor stimulation of PIP2 hydrolysis in parotid gland membranes from aged rats: defect in activation of G alpha q/11.

m3-Muscarinic cholinergic receptor (m3-AChR) and alpha 1-adrenergic receptor (alpha 1-AR) stimulation of phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis (by a PIP2-specific phospholipase C, PLC) in rat parotid gland membranes is mediated via activation of alpha subunits of the Gq/11 family of G-proteins. This study examines m3-AChR and alpha 1-AR stimulation of PIP2 hydrolysis in membranes isolated from parotid glands of old (24 months) and young (3 months) rats (old and young rat membranes). Old rat membranes exhibited reduced stimulation of PIP2 hydrolysis in response to the addition of guanosine-5'-O-(3-thiotrisphosphate) (GTP gamma S) alone or GTP gamma S plus either carbachol (m3-AChR agonist) or epinephrine (alpha 1-AR agonist). This reduction in receptor-stimulated PIP2 hydrolysis was not due to a decrease in PLC activity per se since cholate-solubilized PLC activity was similar in old and young rat membranes. Additionally, these membranes exhibited comparable, immunologically detectable, levels of PLC beta 3, G alpha q/11, and G beta. In the presence of 10 microM AlCl3 and 10 mM NaF, stimulation of PIP2 hydrolysis in both old and young rat membranes was similar. Preincubation of membranes from old rats with GTP gamma S induced a time-dependent increase in the rate of PIP2 hydrolysis and, with 20 min preincubation, the rates of hydrolysis in old and young rat membranes were not statistically different. In aggregate, these data indicate that there is a defect in the activation of G alpha q/11 in parotid gland membranes from old rats.

Existence of gamma-aminobutyric acid and its biosynthetic and metabolic enzymes in rat salivary glands.

To obtain more insight into the physiological role of gamma-aminobutyric acid (GABA) in rat salivary glands, we measured the concentration of GABA and the activities of its biosynthetic and metabolic enzymes, glutamate decarboxylase (GAD) and GABA transaminase (GABA-T). The GABA concentrations in rat parotid and submandibular glands were 10.0 and 14.3 nmol/g weight, respectively, which were 0.6-0.8% of the levels in the brain (cerebellum and medulla oblongata), whereas glutamic acid (Glu) was abundant in the two glands. These GABA levels in the two glands were significantly decreased by administration of semicarbazide (200 mg/kg, i.p.), a GAD inhibitor, and increased by gabaculine (50 mg/kg, i.p.), a GABA-T inhibitor. The activities of both GAD and GABA-T were also detected in homogenates of the two salivary glands, but they were lower than those in the brain. However, kinetic analysis showed that the values of Michaelis constants for Glu and GABA in both enzyme reactions in these two glands were similar to those in the brain. These results indicate that GABA and its biosynthetic and metabolic enzymes are present in rat salivary glands as well as the brain.

Reaction of (1R,2S,3S)-3-methylcyclohexanediamineplatinum(II) with DNA: isolation and characterization of the platinum-nucleotide adducts by means of HPLC and NMR spectroscopy.

Reaction products of calf thymus DNA with (1R,2S,3S)-3-methylcyclohexanediamineplatinum (abbreviated as Pt(RSS-dach)Cl2) were investigated by enzymatic degradation of the platinated DNA and subsequent HPLC analysis. Five platinated adducts involving d(GpG), d(ApG) and (dG)2 residues were identified by HPLC after complete digestion using deoxyribonuclease I, nuclease P1, and alkaline phosphatase. The adducts with d(GpG) and d(ApG) consisted of two geometrical isomers, because Pt(RSS-dach)Cl2 lacks a C2 symmetry element. The d(GpG) and d(ApG) adducts were intrastrand compounds crosslinked between the N7 atoms of the adjacent purine bases. The two d(GpG) adducts were most abundant and comprised more than 65% of all the platinated adducts. The relative ratio of the two d(GpG) isomers was 3:2 for reaction with DNA, whereas the ratio was 1:1 for reaction with a single stranded oligonucleotide. The detailed structure of the two d(GpG) adducts is also described based on NMR spectroscopic data.

Alpha 1-adrenergic and m3-muscarinic receptor stimulation of phosphatidylinositol 4,5-bisphosphate-specific phospholipase C are independently mediated by G alpha q/11 in rat parotid gland membranes.

Stimulation of m3-muscarinic cholinergic receptors (m3AChR) in the rat parotid gland increases the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) via activation of the G alpha/11 family of G-proteins (Sawaki et al. (1993) Arch. Biochem. Biophys., 546-550). Herein we report that alpha 1-adrenergic receptor (alpha 1-AR) stimulation of PIP2 hydrolysis is also mediated via alpha subunits of the G q/11 family of G-proteins. The alpha 1-AR agonist, epinephrine, induced a dose-dependent increase (1.5-fold maximum) of exogenously added PIP2 in the presence of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S), which was inhibited by the alpha 1-AR antagonist, phentolamine, but not by the beta-adrenergic receptor antagonist, propranolol. The epinephrine-stimulated component of PIP2 hydrolysis was significantly inhibited by pretreating the membranes with an antiserum against G alpha q/11. When carbachol and epinephrine were present simultaneously (with GTP gamma S), the increase in PIP2 hydrolysis obtained was not significantly different from the sum of the increases in PIP2 hydrolysis obtained with each agonist alone. PIP2 hydrolysis stimulated in the presence of carbachol and epinephrine was inhibited by alpha 1-AR and m3AChR antagonists, phentolamine and atropine respectively, to the level obtained with each agonist alone. Notably, in the presence of both agonists the inhibition of PIP2 hydrolysis by anti-G alpha q/11 antiserum was not significantly different from the sum of the inhibitions obtained with each agonist alone. These results indicate that m3AChR and alpha 1-AR stimulation of PIP2 hydrolysis in rat parotid gland membranes are independently mediated by the G alpha q/11 family of G-proteins.

Involvement of G alpha q/11 in m3-muscarinic receptor stimulation of phosphatidylinositol 4,5 bisphosphate-specific phospholipase C in rat parotid gland membranes.

Carbachol stimulation of phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis by rat parotid gland membranes is dependent on the presence of GTP gamma S and is a result of m3-muscarinic receptor regulation of G-protein coupled, PIP2-specific phospholipase C (PLC). The PLC activity (> 80%) was solubilized with 1% Na-cholate but the solubilized enzyme was not stimulated by GTP gamma S and carbachol. Immunoblotting of rat parotid membranes with polyclonal antiserum, which recognizes alpha-subunits of the Gq/11 family, indicated the presence of two immunoreactive proteins of approximate molecular weights 41 and 42 kDa. Incubation of membranes with the common G alpha q/11 antiserum attenuated the stimulation of PIP2 hydrolysis, induced by GTP gamma S alone and by carbachol, in the presence of GTP gamma S. The antiserum had no effect on PIP2 hydrolysis in unstimulated membranes or in the cholate extract, where it is uncoupled from the G-protein. Antiserum against G alpha i, which is also coupled to the m3-muscarinic receptor in this tissue, had no effect on either basal or stimulated PIP2 hydrolysis. These results demonstrate that in rat parotid gland, activation of PIP2-specific PLC by m3-muscarinic receptor stimulation is mediated via alpha-subunits of the Gq/11 family of G-proteins.

Effects of kampo (Japanese herbal) medicine "sho-saiko-to" on DNA-synthesizing enzyme activity in 1,2-dimethylhydrazine-induced colonic carcinomas in rats.

Sho-Saiko-To (SST) is a modified Japanese traditional Chinese herbal medicine containing seven medical plants: Bupleuri radix, Pinelliae tuber, Suxtallariae radix, Zizyphi fructus, Ginseng radix, Glycyrrhizae radix, and Zingiberis recens rhizoma. This preparation has been used in the treatment of some inflammatory diseases of the respiratory system and chronic hepatitis. In the present study, the effects of SST were investigated on the activities of DNA-synthesizing enzymes in 1,2-dimethylhydrazine (DMH)-induced colonic carcinomas in rats. Six-week administration of SST prevented nearly 100% of the body weight loss and the final number of the colonic carcinomas compared to those in the rats treated with DMH alone, and suppressed the enhanced activities of thymidylate synthetase (TS) and thymidine kinase (TK) which were involved in the de novo and salvage pathways of pyrimidine synthesis, respectively, in DMH-induced colonic carcinomas. These results indicate that SST may show directly and/or indirectly inhibitory effects on the development of colonic carcinomas.

Effects of danazol on DNA synthesis in rat prostate.

Effects of danazol, an isoxazol derivative of the synthetic steroid 17 alpha-ethinyltestosterone, on activities of thymidylate synthetase and thymidine kinase, which are the DNA-synthesizing enzymes included in de novo and salvage pathways of pyrimidine metabolism, respectively, were investigated in rat prostate. Danazol markedly reduced plasma levels of luteinizing hormone and testosterone, and organ weight, both enzyme activities and bromodeoxyuridine-immunoreactive cells which were regarded as the S-phase cells in prostate. These results indicate that danazol shows a property as a potent antigonadotropin.

Effects of luteinizing hormone-releasing hormone analogue on DNA synthesis in rat prostate and uterus.

Buserelin, a potent LH-RH agonist, has been used for the treatment of hormonal disorders such as precocious puberty, endometriosis, cystic mastitis and prostatic carcinoma. Prolonged treatment with buserelin has been known to induce a refractory phase of pituitary desensitization. In the present study, we found that two-week treatment with buserelin strongly suppressed the activities of thymidylate synthetase and thymidine kinase, and markedly reduced the appearance of BrdU-immunoreactive (S-phase) cells in both prostate glands and uteri in male and female adult rats, respectively.