The differential expression of mgl mRNA by Porphyromonas gingivalis affects the production of methyl mercaptan.

OBJECTIVE: A large number of individuals have halitosis. The total amount of volatile sulfur compounds, which are the main cause of halitosis, has been correlated with periodontitis following bacterial infection. In this study, Porphyromonas gingivalis (Pg), a major periodontopathogenic bacterium, was isolated from patients with halitosis by the amplification of 16S rRNA, and the ability of isolated Pg to produce methyl mercaptan (CH3 SH) was determined to clarify the relationship between halitosis and Pg infection. MATERIALS AND METHODS: CH3 SH concentrations were measured in patients using Oral Chroma. The production of CH3 SH by Pg standard and clinical strains was also measured in vitro. Real-time PCR was performed to compare the expression of mgl mRNA (which encoded l-methionine-a-deamino-g-mercaptomethane-lyase) among the Pg strains. The production of CH3 SH and the expression of mgl mRNA were also determined to assess the effects of oriental medicine. RESULTS: The production of CH3 SH and the expression of mgl mRNA strongly correlated with each other in the presence of l-methionine. The expression of mgl mRNA by Pg W83 was strongly inhibited by magnoliaceae. CONCLUSION: The production of CH3 SH was correlated with the expression of mgl. Furthermore, the oriental medicine, magnoliaceae, may represent a potential treatment for halitosis.

Relationship between xerostomia and gingival condition in young adults.

BACKGROUND AND OBJECTIVE: Xerostomia is a subjective symptom of dryness in the mouth. Although a correlation between xerostomia and oral conditions in the elderly has been reported, there are few such studies in the young adults. The aim of this study was to examine the relationship of xerostomia with the gingival condition in university students. MATERIAL AND METHODS: A total of 2077 students (1202 male subjects and 875 female subjects), 18-24 years of age, were examined. The disease activity and severity of the gingival condition were assessed as the percentage of teeth with bleeding on probing (%BOP) and the presence of teeth with probing pocket depth of ≥ 4 mm, respectively. Additional information on xerostomia, oral health behaviors, coffee/tea intake and nasal congestion was collected via a questionnaire. Path analysis was used to test pathways from xerostomia to the gingival condition. RESULTS: One-hundred and eighty-three (8.8%) students responded that their mouths frequently or always felt dry. Xerostomia was related to %BOP and dental plaque formation, but was not related to the presence of probing pocket depth ≥ 4 mm. In the structural model, xerostomia was related to dental plaque formation (p < 0.01), and a lower level of dental plaque formation was associated with a lower %BOP. Xerostomia was associated with coffee/tea intake (p < 0.01) and nasal congestion (p < 0.001). CONCLUSION: Xerostomia was indirectly related to gingival disease activity through the accumulation of dental plaque. Nasal congestion and coffee/tea intake also affected xerostomia. These findings suggest that xerostomia should be considered in screening for gingivitis risk in young adults.

Involvement of regulatory elements on corticotropin-releasing factor gene promoter in hypothalamic 4B cells.

INTRODUCTION: Corticotropin-releasing factor (CRF) plays a central role in controlling the hypothalamic-pituitary-adrenal (HPA) axis during stressful periods. CRF is synthesized and secreted in the hypothalamic paraventricular nucleus (PVN) in response to stress, and stimulates ACTH in the pituitary corticotrophs. ACTH stimulates the release of glucocorticoids from the adrenal glands, and glucocorticoids sequentially inhibit hypothalamic PVN production of CRF and pituitary production of ACTH. The effects of glucocorticoids on CRF gene regulation, however, are possibly tissue-specific since glucocorticoids stimulate CRF gene expression in the placenta and the bed nucleus of the stria terminalis, while they inhibit it in the hypothalamus. METHODS AND RESULTS: In a hypothalamic cell line, 4B, we found that forskolin-stimulated CRF gene transcription was mediated by a functional cAMP-response element (CRE), which included -220 to -233 bp on the CRF 5'-promoter region. Protein kinase A, protein kinase C, and p38 mitogen-activated protein kinase pathways contributed to forskolin-induced transcriptional activity of CRF in hypothalamic 4B cells. Glucocorticoid-dependent repression of cAMP-stimulated transcriptional activity of CRF was localized to promoter sequences between -278 and -233 bp, which included a glucocorticoid regulatory element and a serum response element. CONCLUSION: Taken together, these findings indicate that the regulatory elements, including CRE, negative glucocorticoid regulatory element, and a serum response element on the promoter, contribute to the regulation of CRF gene transcription in hypothalamic 4B cells.

Impaired parathyroid hormone response to hypocalcemic stimuli in a patient with hypomagnesemic hypocalcemia.

Magnesium (Mg) deficiency sometimes causes hypocalcemia with impaired PTH secretion although the precise mechanism remains unclear. We examined the PTH secretion in response to physiological hypocalcemic stimuli in a patient with hypomagnesemic hypocalcemia. We adopted sodium bicarbonate infusion test, which we recently developed, to evaluate the PTH response to acute decrease in plasma ionized Ca. The results showed that, before Mg replacement and when the patient was mildly hypocalcemic, absolutely no PTH release to hypocalcemic stimuli was observed. In contrast, the plasma Ca was promptly normalized following the start of Mg replacement, and brisk PTH response to hypocalcemic stimuli was obtained during the same test carried out a week after the Mg replacement. The data in this case thus suggest that: a) the acute regulation of PTH release by plasma ionized Ca is lost in the patient with hypomagnesemic hypocalcemia, and b) Mg deficiency itself is likely to be a primary cause of this disorder because the hormone response was clearly restored after shortterm Mg replacement alone.

Suppressive effects of dehydroepiandrosterone and the nuclear factor-kappaB inhibitor parthenolide on corticotroph tumor cell growth and function in vitro and in vivo.

Dehydroepiandrosterone (DHEA) is believed to have an anti-tumor effect, as well as anti-inflammatory, antioxidant, and anti-aging effects. To clarify the possible inhibitory action of DHEA on pituitary tumor cells, we tested the effects of DHEA, alone or in combination with the nuclear factor-kappaB (NF-kappaB) inhibitor parthenolide (PRT), on AtT20 corticotroph cell growth and function both in vitro and in vivo. We found that, in vitro, DHEA and PRT had potent inhibitory effects on pro-opiomelanocortin and NF-kappaB-dependent gene expression. They also suppressed the transcription activity of survivin, a representative anti-apoptotic factor, and induced apoptosis in this cell line. Furthermore, using BALB/C nude mice with xenografts of AtT20 cells in vivo, we found that the combined administration of DHEA and PRT significantly attenuated tumor growth and survivin expression. The treatment also decreased the elevated plasma corticosterone levels and ameliorated the malnutrition induced by tumor growth. Altogether, these results suggested that combined treatments of DHEA and PRT potently inhibit the growth and function of corticotroph tumor cells both in vitro and in vivo. This effect may, at least partly, be caused by the suppressive effects of these compounds, such as survivin and other inhibitor of apoptosis proteins, on NF-kappaB-mediated gene transcription.

Regulatory mechanisms of corticotropin-releasing hormone and vasopressin gene expression in the hypothalamus.

Tuberoinfundibular corticotropin-releasing hormone (CRH) neurones are the principal regulators of the hypothalamic-pituitary-adrenal (HPA)-axis. Vasopressin is primarily a neurohypophysial hormone, produced in magnocellular neurones of the hypothalamic paraventricular and supraoptic nuclei, but parvocellular CRH neurones also coexpress vasopressin, which acts as a second 'releasing factor' for adrenocorticotropic hormone along with CRH. All stress inputs converge on these hypothalamic neuroendocrine neurones, and the input signals are integrated to determine the output secretion of CRH and vasopressin. Aminergic, cholinergic, GABAergic, glutamatergic and a number of peptidergic inputs have all been implicated in the regulation of CRH/vasopressin neurones. Glucocorticoids inhibit the HPA-axis activity by negative feedback. Interleukin-1 stimulates CRH and vasopressin gene expression, and is implicated in immune-neuroendocrine regulation. cAMP-response element-binding protein phosphorylation may mediate transcriptional activation of both CRH and vasopressin genes, but the roles of AP-1 and other transcription factors remain controversial. Expression profiles of the CRH and vasopressin genes are not uniform after stress exposure, and the vasopressin gene appears to be more sensitive to glucocorticoid suppression.

Reflex sympathetic dystrophy following transbrachial cardiac catheterization.

We experienced a rare case of complication by reflex sympathetic dystrophy (RSD) following transbrachial cardiac catheterization which may have been caused by poorly executed hemostasis using a hemostatic device. The symptoms of RSD markedly limited the patientOs daily work activities. Although the transbrachial approach is a useful procedure for cardiac catheterization, interventionalists should be aware that RSD may cause serious complications.

Metabolism of 1-acyl-2-oleoyl-sn-glycero-3-phosphoethanolamine in castor oil biosynthesis.

We have examined the role of 2-oleoyl-PE (phosphatidylethanolamine) in the biosynthesis of triacylglycerols (TAG) by castor microsomes. In castor microsomal incubation, the label from 14C-oleate of 1-palmitoyl-2-[1-(14)C]oleoyl-sn-glycero-3-phosphoethanolamine is incorporated into TAG containing ricinoleate. The enzyme characteristics, such as optimal pH, and the effect of incubation components of the oleoyl-12-hydroxylase using 2-oleoyl-PE as incubation substrate are similar to those for 2-oleoyl-PC (phosphatidylcholine). However, compared to 2-oleoyl-PC, 2-oleoyl-PE is a less efficient incubation substrate of oleoyl-12-hydroxylase in castor microsomes. Unlike 2-oleoyl-PC, 2-oleoyl-PE is not hydroxylated to 2-ricinoleoyl-PE by oleoyl-12-hydroxylase and is not desaturated to 2-linoleoyl-PE by oleoyl-12-desaturase. We have demonstrated the conversion of 2-oleoyl-PE to 2-oleoyl-PC and vice versa. The incorporation of label from 2-[14C]oleoyl-PE into TAG occurs after its conversion to 2-oleoyl-PC, which can then be hydroxylated or desaturated. We detected neither PE-N-monomethyl nor PE-N,N-dimethyl, the intermediates from PE to PC by N-methylation. The conversion of 2-oleoyl-PE to 2-oleoyl-PC likely occurs via hydrolysis to 1,2-diacyl-sn-glycerol by phospholipase C and then by cholinephosphotransferase. This conversion does not appear to play a key role in driving ricinoleate into TAG.

Autologous blood donation with recombinant human erythropoietin for abdominal aortic aneurysm surgery.

We evaluated the efficacy of autologous blood donation using recombinant human erythropoietin for elective abdominal aortic aneurysm (AAA) surgery regarding postoperative recovery. Twenty-five AAA patients who completed surgery without receiving a homologous blood transfusion were divided into two groups, consisting of a control group (n = 12) who did not bank any autologous blood, and a donation group (n = 13) who did bank more than 800 ml of autologous blood with the use of erythropoietin. The hematocrit (Ht) level of the control group decreased from 41.1% +/- 1.2% before the operation to 36.2% +/- 0.9% just afterwards (P > 0.01). In the donation group, however, the Ht did not change significantly during either the donation period or the perioperative period. The postoperative period before oral food intake and natural defecation were both significantly shorter in the donation group than in the control group. The first day of mobilization was also earlier in the donation group. In conclusion, autologous blood donation using erythropoietin for AAA surgery is therefore considered to promote the early recovery of patients.

T-588, a novel neuroprotective agent, delays progression of neuromuscular dysfunction in wobbler mouse motoneuron disease.

R(-)-1-(benzo[b]thiophen-5-yl)-2-[2-(N,N-diethylamino) ethoxy]ethanol hydrochloride (T-588) enhances acetylcholine release from the frontal cortex and hippocampus in rats, and can ameliorate cognitive dysfunction in various amnesia models of rodents. T-588 protects rat cerebellar granule cells from glutamate neurotoxicity in culture. This agent also inhibits facilitation in the crayfish neuromuscular junction and mammalian cerebellum. Clinical trials of T-588 are underway in patients with Alzheimer's disease. We attempted to determine whether T-588 treatment ameliorates neuromuscular dysfunction in the wobbler mouse, an animal model of motoneuron disease (MND). After the initial diagnosis of MND at the age of 3-4 weeks, wobbler mice were orally administered T-588 (3, 10, 30 mg/kg) or vehicle daily for 4 weeks in a blinded fashion. We compared symptomatic, pathological and biochemical changes among the groups. In comparison with vehicle, T-588 administration potentiated grip strength, attenuated forelimb contracture and increased the weight of the biceps muscles. T-588-treated mice had retarded denervation muscle atrophy and elevated activities of choline acetyltransferase (ChAT) or lactate dehydrogenase in the biceps muscles. T-588 treatment also enhanced ChAT activities and promoted formation of cyclic adenosine monophosphate in the cervical cord. Pharmacokinetic study also showed that T-588 was transported efficiently into the cerebrum and spinal cord following oral administration. Thus, T-588 treatment delayed the progression of wobbler murine MND. Our findings suggest that this agent has therapeutic potential in human motor neuropathy or MND.

[Preoperative laparoscopy by local anesthesia for advanced gastric cancer].

Preoperative laparoscopy by local anesthesia was performed in 8 patients with advanced gastric cancer, whose lesions had been diagnosed to be more than T3 or suspected to have peritoneal seeding, and its usefulness was assessed. After insertion of the trocars, the abdominal cavity was inspected, and biopsy and/or abdominal lavage sampling was performed. Three patients out of 8 were diagnosed as P3, and 5 patients were diagnosed as P0 and CY0. Based on these results, 6 patients underwent operation. The accuracy rate of diagnosis was 83% in P category, and 100% in CY category. In conclusion, it is considered that laparoscopy by local anesthesia is a useful preoperative examination for advanced gastric cancer.

[A case of type 4 gastric cancer with peritoneal dissemination treated with intra-aortic chemotherapy].

A 45-year-old woman was admitted to our hospital because of lower abdominal pain and anorexia. A barium gastrography and gastroscopy showed a type 4 gastric cancer in the upper gastric body. Histologic study on biopsy specimens from the tumor revealed poorly differentiated adenocarcinoma. Computed tomography revealed bilateral hydronephrosis, and barium enema showed diffuse stenosis of the sigmoid colon because of peritoneal dissemination. This patient was treated by intra-aortic infusion therapy with sequential MTX and 5-FU. After five courses of the administration, barium enema revealed reexpansion of the lumen of sigmoid colon with normalization of the tumor markers. The patient was discharged without symptoms. Intra-aortic infusion therapy with sequential MTX and 5-FU was considered an effective treatment for unresectable gastric cancer.

Neuronal nitric oxide synthase inhibitor, 7-nitroindazole, delays motor dysfunction and spinal motoneuron degeneration in the wobbler mouse.

Gene mutations of superoxide dismutase (SOD) have been discovered in familial amyotrophic lateral sclerosis (ALS). Neuronal nitric oxide synthase (NOS), endothelial NOS and 3-nitrotyrosine immunoreactivities are selectively increased in the spinal motoneurons of sporadic ALS. Other study suggests that 3-nitrotyrosine immunoreactivity is enhanced in the spinal motoneurons of sporadic and familial ALS patients. The hypothesis is postulated that increased production of radical species, such as superoxide and peroxynitrite, may cause motoneuron degeneration in ALS. There are increased amounts of nitric oxide and SOD hypoactivities in the brain and spinal cord of wobbler mice. NOS is also induced in the vacuolated spinal motoneurons or axons in this animal. Free radicals might contribute to the pathogenesis of wobbler mouse motoneuron disease. Lecithinized SOD treatment has retarded the progression of this disease. This evidence allowed us to determine whether NOS inhibitors delay progression of wobbler mouse motoneuron disease. After clinical diagnosis at age 3-4 weeks, wobbler mice were injected with intraperitoneal non-selective NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME, 50 mg/kg), two doses of neuronal NOS inhibitor, 7-nitroindazole (5 or 50 mg/kg) or a vehicle solution, daily for 4 weeks in a blind fashion. In comparison with vehicle, 7-nitroindazole-treated mice potentiated grip strength and attenuated deformities in the forelimbs. 7-Nitroindazole treatment increased the biceps muscle weight, reduced denervation muscle atrophy, and suppressed degeneration of spinal motoneurons. To a lesser degree, L-NAME-treated mice displayed slowed progression of disease. The present studies indicate that neuronal NOS inhibitor may be a candidate for promising therapy in lower motoneuron disease or motor neuropathy.

Interpretation of accelerants in blood of cadavers found in the wreckage after fire.

Accelerants in the blood of 73 cadavers found in wreckage after fire were analyzed by gas chromatography (GC) and a combination of gas chromatography-mass spectrometry (GC-MS) to decide whether accelerants containing petroleum components had been used and whether the cadavers had been exposed to fire before or after death. In 16 of 26 cases in which accelerants were used to start a fire before death, accelerants were detected in the blood. In 7 cases in which accelerants were used to start a fire, the victims were determined to have been exposed to the vapor of accelerants after death because no accelerants were detected in the blood, no soot was found in the airways, and carboxyhemoglobin (COHb) concentrations were not higher than those found in smokers. In 9 of 34 cases in which accelerants were suspected to have been used to start a fire before death, accelerants were detected in the blood. When soot is not detectable by the unaided eye in the airways of a victim found in debris of a fire in which the use of accelerants is suspected, or the COHb concentration in the blood is no higher than in a smoker, analysis of accelerants in the blood seems to be helpful in determining the cause of death and whether inflammable were used.

[Evaluation of clinical trials on adjuvant chemotherapy for curatively resected gastric cancer].

Reports of the results of clinical trials on adjuvant chemotherapy for curatively resected gastric cancer in Japan and other countries were reviewed. The efficacy of adjuvant chemotherapy outside Japan was not recognized except in two trials. Japanese trials did not reveal a significant survival benefit compared with surgery alone. However, a consensus was achieved in Japan on its efficacy by understanding some results of subset analysis as definite proof. Recently, it has been recognized that the re-evaluation of adjuvant chemotherapy should be carried out with the control group of surgery alone. The results of the trials of JCOG and N.SAS-GC, comparing adjuvant chemotherapy versus surgery alone, are much awaited. At the present time, carefully designed randomized controlled trials with sufficient sample size, which include surgery alone for control arm, are the only way to establish the efficacy of adjuvant chemotherapy.

Metabolic and neurological patterns in chronic cerebral infarction: a single-voxel 1H-MR spectroscopy study.

The details of brain metabolism in chronic cerebral infarcts have not been clarified. Using proton MR spectroscopy (1H-MRS) at 1.5 T, we measured biochemical changes in 16 patients with large infarcts involving the motor cortex in the chronic phase (median 293.9 days) and related the findings to clinical data. Localised spectra were obtained using point-resolved spectroscopy, with an echo time of 270 ms. Regions of interest were placed on the frontal lobe, including the precentral gyrus and central sulcus. Motor function was assessed by the manual muscle power test at the time of the 1H-MRS study. Only three patients with severe paresis had no signal in the lesions and a lactate signal was obtained in 13 cases. N-acetyl aspartate (NAA) was observed in 4 cases with recanalisation of an occluded vessel. Motor function correlated strongly with the NAA/choline-containing compounds (Cho) ratio (P < 0.01) and lactate/Cho ratio (P < 0.01). We found various metabolic patterns, reflecting residual neurological function.

Effect of transforming growth factor beta 1 on spinal motor neurons after axotomy.

Glial cell line-derived neurotrophic factor (GDNF), a member of the transforming growth factor beta (TGF-beta) family, has potent effects on developing motor neurons. TGF are pluripotent cytokines that exert biological effects on a variety of neurons. TGF beta 1, on the other hand, promotes motor neuron survival in vitro and saves motor neurons from naturally occurring cell death. Here we investigate the neurotrophic effects of TGF beta 1 for axotomized motor neuron death. The sciatic nerve was cut in newborn rats and TGF beta 1 was injected, either by intraperitoneally or by lesion site, for 14 days after transection. Two or six weeks postlesion, the number and the diameter of motor neurons was assessed. TGF beta 1 significantly attenuated axotomy induced motor neuron death by intraperitoneal administration or by lesion site administration at 2 weeks after neonatal axotomy in a similar way. However, no effect was observed at 6 weeks after nerve lesion, despite continuous application of TGF beta 1 daily for 14 days. These results indicate that TGF beta 1 can prevent the death of motor neurons in vivo, but it cannot permanently rescue lesioned motor neurons.

Centrally administered neuropeptide FF inhibits arginine vasopressin release in conscious rats.

There is evidence indicating that neuropeptide FF (NPFF) is an endogenous modulator of opioid systems. In the present study, we investigated the effect of centrally administered NPFF on arginine vasopressin (AVP) release in conscious rats. The plasma AVP increase in response to either hyperosmolality [i.p. injection of hypertonic saline (600 mosmol/kg)] or hypovolemia [i.p. injection of polyethylene glycol (PEG)] was significantly blunted when NPFF was injected into the lateral ventricle so that the given drug could act at the hypothalamus and also reach the brain stem (hypertonic saline with 10 micrograms/rat NPFF, 3.28 +/- 0.48 pg/ml; hypertonic saline alone, 7.85 +/- 1.78 pg/ml; PEG with 10 micrograms/rat NPFF, 4.07 +/- 1.40 pg/ml; PEG alone, 8.25 +/- 1.90 pg/ml). The plasma AVP increase in response to PEG-induced hypovolemia was also attenuated significantly and more potently when NPFF was injected into the cisterna magna so that the given drug could be readily accessible to the dorsal medulla where the nucleus of solitary tract is located (10 micrograms/rat; 2.71 +/- 0.14 pg/ml). In contrast, the NPFF injected into the cisterna magna had no significant effect on hyperosmolality-induced AVP release. Treatment with naloxone (10 mg/kg BW, sc) significantly reversed the inhibitory effects of NPFF on AVP release. These results suggest that central NPFF might play an inhibitory role via the hypothalamus in the osmoregulation of plasma AVP and via both the hypothalamus and the nucleus of solitary tract in the baroregulation, and that the intrinsic opioid systems are involved in the action of NPFF.