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.

[High-pressure suction drainage for poststernotomy mediastinitis].

We present a case of high-pressure suction drainage for poststernotomy mediastinitis. A 67-year-old man who underwent coronary artery bypass grafting because of angina pectoris was noted massive dirty exudate fluid (putrid secretions) from median sternal wound on the 6th postoperative day. The sternal wounds were completely reopened and all sternal wires removed. Thorough debridement was performed. The mediastinum was then washed out with warn normal saline. The polyurethane foam was shaped to fit the resulting sternal defect and placed within the cavity just below the skin edge. The suction tube was inserted inside the foam and the area was covered with the adhesive drape. After 45 days suction drainage the sternal wound was closed, after which he was discharged.

Urinary trypsin inhibitor suppresses vascular smooth muscle contraction by inhibition of Ca2+ influx.

Urinary trypsin inhibitor (UTI) and its precursor form inter-alpha trypsin inhibitor (ITI) are present in plasma. To determine the action of UTI on blood vessels, we performed isometric vascular muscle contraction tests, microcirculation studies and measurement of cytosolic free Ca2+ in vascular smooth muscle cells. An isometric vascular muscle contraction test showed that the contractions stimulated by endothelin-1 or norepinephrine were suppressed in the presence of UTI, and that the contractions were not inhibited in the presence of ITI. The microcirculation study showed that the contraction of mesenteric arterioles of WKY rats induced by norepinephrine were inhibited by treatment of UTI, and that they did not alter by treatment of ITI. Pre-incubation of UTI, but not ITI, with vascular smooth muscle cells inhibited the increase of cytosolic free Ca2+ induced by endothelin-1 or norepinephrine. Cell-binding study by biotinylated UTI showed that vascular smooth muscle cells have specific binding site for UTI, but not for ITI. We propose that circulating UTI converted from ITI has a regulatory effect on local vascular tone by regulation of Ca2+ influx into smooth muscle cells.

Direct expression in Escherichia coli and characterization of bovine adrenodoxins with modified amino-terminal regions.

Four forms of bovine adrenodoxin with modified amino-termini obtained by direct expression of cDNAs in Escherichia coli are Ad(Met1), Ad(Met-1), Ad(Met-12), and Ad(Met6). The shoulder numbers represent the site of translation initiator Met at the amino-termini. The adrenodoxins, except for Ad(Met-1), were purified from the cell lysate and the ratios of A414-to-A276 of the purified proteins were over 0.92. NADPH-cytochrome c reductase activities of the three forms of adrenodoxin in the presence of adrenodoxin reductase were the same as that of purified bovine adrenocortical adrenodoxin. However, as cytochrome P-450SCC reduction catalyzed by Ad(Met6) was about 60% of that by Ad(Met1), the contribution of the amino-terminal region for the electron transfer or binding to cytochrome P-450SCC would need to be considered.

The impact of vasoactive peptides on nitric oxide production in cultured sympathetic neurons.

The concentration of nitric oxide was found to be decreased in a hypersympathetic condition. We carried out experiments on cultured sympathetic neurons from 12-14-days-old chick embryos to investigate the role of vasoactive peptides and amine on nitric oxide production. Stimulation of cultured neurons with endothelin-1, norepinephrine and angiotensin-II initially increases nitric oxide production and subsequently decreases it in a dose-dependent manner (P<0.05, n = 7). Stimulation of Fura-2 acetoxymethyl ester-loaded neurons with endothelin-1, norepinephrine and angiotensin-II increases the calcium influx (within 30-90 s) and it is then restored to the initial level (P<0.05, n = 7). An additional observation was that specific stimulator L-arginine significantly increases the nitric oxide release and calcium influx into the cells, whereas N(W)-nitro-L-arginine methyl ester blunts nitric oxide release dose dependently (P<0.05, n = 7) and does not change the calcium concentration in the cells. We propose that vasoactive peptides and amines inhibit nitric oxide production in the cultured sympathetic neuron by regulation of intracellular calcium concentration.

Induction of hypercoagulability condition by chronic localized cold stress in rabbits.

To evaluate the effect of cold-induced stress on renal and hepatic blood flow and coagulation parameters, rabbits' soles were exposed to ice pad (0 degrees C). Renal and hepatic blood flow was measured after 1 h and 15 days of cold stress. Coagulation parameters (0, 8th and 15th days of stress) and histological studies were performed. Renal and hepatic blood flow was significantly reduced after cold-stress. Decreased platelet count, antithrombin III (AT III) activity, increased fibrinogen (Fbg) level, shortened activated partial thromboplastin time (aPTT) and prothrombin time (PT) was found after 8 and 15 days of cold-stress. Histology showed enlarged glomeruli with fibrin deposition in kidney, ischemic changes and fibrin deposition in liver and hemorrhagic necrosis in adrenal cortex. We conclude that undesirable localized cold induced sympathetic stimulation in daily life may be a predisposing factor for coagulopathy.

Subcutaneous and mediastinal emphysema associated with hypersensitivity pneumonitis.

We report a rare case of a patient in whom severe subcutaneous and mediastinal emphysema occurred in association with summer-type hypersensitivity pneumonitis and in whom overdistention or disruption of alveoli with obliteration of the respiratory bronchioles was revealed on open lung biopsy. This case suggests that obstructive bronchiolitis with hypersensitivity pneumonitis is an etiologic factor of mediastinal emphysema.

Neutrophilia in bronchoalveolar lavage fluid of diffuse panbronchiolitis.

The clinical and pathologic features of diffuse panbronchiolitis (DPB) have been well reported to date, although its pathogenesis remains unknown. In the present study, we performed bronchoalveolar lavage (BAL) on three patients with biopsy specimen-proven DPB and eight patients with highly probable DPB (six women and five men; one smoker and ten nonsmokers), nine patients with chronic bronchitis (all men, five smokers and four nonsmokers), and nine normal control subjects (six women and three men, all nonsmokers) to clarify the cell populations in the lower respiratory tract. Neutrophils comprised 55.3 +/- 24.4 percent of recovered cells by BAL in DPB patients but only 6.6 +/- 6.4 percent in chronic bronchitis patients, and 1.8 +/- 1.5 percent in normal control subjects (p less than 0.001, all comparisons). The DPB patients also exhibited a decreased percentage of alveolar macrophages (34.9 +/- 23.5 percent) compared with chronic bronchitis patients and normal control subjects (p less than 0.001, all comparisons). The percentage of lymphocytes of the recovered lavage cells in DPB patients did not differ from that in chronic bronchitis patients and normal control subjects. These results indicate that neutrophils play an important role in the pathogenesis of DPB. They also suggest that neutrophilia of BAL-recovered fluid is a common finding in diseases associated with bronchiolar inflammation despite some clinical and pathophysiologic differences.

Anemia in patients with colorectal cancer.

Although anemia is one of the signs of colorectal cancer, the relationships between histological findings and hematological findings other than hemoglobin level have not been adequately investigated. We investigated the relationship between hematological findings, serum iron, and histological findings in 358 patients (207 men and 157 women) with colorectal cancer. Their mean (+/-SD) ages were 64.3 +/- 12.4 and 63.8 +/- 13.3 years. A hemoglobin level of less than 10 g/dl was the criterion for anemia, and 20.8% of the men and 25.8% of the women met this criterion. Univariate analysis showed that carcinoma of the cecum, ascending colon, and transverse colon; large-size carcinoma, invasion beyond the proper muscle layer; positive lymph node metastasis: and clinical stage (Dukes' B, C, and D) were factors associated with high incidence of anemia. Histological type did not affect the hematological findings. Multivariate analysis showed that age, tumor site, and tumor size were significant factors related to anemia. Depth of invasion, the presence or absence of lymph node metastasis, and Dukes' classification were not significant factors. In the presence of these factors, mean corpuscular volume and mean corpuscular hemoglobin concentration values were low, and red blood cells were microcytic and hypochromic. The incidence of a low serum iron level was about twice the frequency of a hemoglobin level of less than 10 g/dl. The results of the multivariate analysis showed that none of the factors were significantly related to iron deficiency.

A new treatment of severe pre-eclampsia by long-term epidural anaesthesia.

We hypothesised that long term epidural anaesthesia suppresses sympathetic overactivity in pre-eclampsia. Two equal groups of severe pre-eclamptic patients were treated either with long-term epidural anaesthesia (epidural group) or bed rest, diet control, and an antihypertensive drug (control group). After 7 days of epidural block, mean arterial blood pressures decreased, platelet count and serum total protein increased in all cases while proteinuria decreased in four cases. All patients treated with a long-term epidural therapy continued their pregnancies for more than 3 weeks after admission. The infants in the epidural group had 1-min Apgar scores above 8, a body weight of 2240+/-310 g (mean +/- s.d.) and normal neonatal progress. In contrast, the pregnancies of eight patients in the control group were terminated within 2 weeks of admission due to severe pre-eclampsia or foetal distress, the birth weight of the infants was 1590+/-380 g (mean +/- s.d.) and four had neonatal distress. Progressive worsening in the mean arterial pressure, proteinuria, platelet count and serum total protein was found in these patients. Long-term epidural anaesthesia suppresses the sympathetic hyperactivity and thus improves pre-eclamptic condition which may open a new treatment in case of progressive severe pre-eclampsia.

Preoperative evaluation by whole-body 18F-fluorodeoxyglucose positron emission tomography in patients with primary colorectal cancer.

Whole-body positron emission tomography (WB PET) was performed preoperatively in 24 patients with primary colorectal cancer, and the results were compared with the histopathological findings. The PET positive rate was 95.8% (23/24 patients) for the primary tumor. Among nine patients with histologically confirmed lymph node metastasis, two (22.2%) were positive by PET, including one with n1 and one with n4 disease. Among 15 patients without lymph node metastasis histopathologically (n0), two (13.3%) showed false-positive nodes on PET, being diagnosed as n1 and n3 disease, respectively. These results suggest that preoperative PET is useful for the diagnosis of primary colorectal cancer, but it is of limited value for detecting metastasis to the regional lymph nodes surrounding the primary lesion.

The expression of MMP-2 and TIMP-2 in patients with primary colorectal adenocarcinoma: correlation with liver metastasis.

The expression of MMP-2 and TIMP-2 was examined immunohistochemically in a total of 36 colorectal cancer patients with synchronous liver metastasis at the time of surgery, and the serum MMP-2 and TIMP-2 levels were also measured in a total of 58 colorectal cancer patients with/without liver metastasis following primary colorectal surgical resection for serological comparison. Although MMP-2 exhibited a significant expression immunohistochemically in the primary colorectal cancer (p<0.05) and TIMP-2 in the synchronous liver metastasis (p<0.01), there was no relationship between the absence/presence of liver metastasis and serum MMP-2 and TIMP-2 levels, respectively. These results suggest that it can be considered difficult to use serum MMP-2 and TIMP-2 levels to predict the status of liver metastasis following primary resection in patients with colorectal adenocarcinoma.

Long-term survival after immunochemotherapy for juvenile colon cancer with peritoneal dissemination: a case report.

A 20 year-old man was hospitalized with an abdominal mass and abdominal distension. Investigations resulted in a diagnosis of ileus caused by advanced colon cancer with peritoneal dissemination to the pouch of Douglas. Palliative surgery was performed to relieve bowel obstruction and debulk the tumor. Histopathological examination showed that the tumor was a mucinous adenocarcinoma invading the serosa without lymph node metastasis. Ascites collected during the operation was diagnosed as class V. Administration of PSK (3.0 g/day) and UFT (600 mg/day) as adjuvant immunochemotherapy was started postoperatively to achieve tumor dormancy. He has been followed as an outpatient for 2.5 years with no ascites or abdominal symptoms.

The expression of MMP-2 and TIMP-2 in patients with colorectal adenocarcinoma invaded to the submucosal and proper muscle layer.

The expression of MMP-2 and TIMP-2 was examined immunohistochemically in a total of 87 colorectal cancer patients. The expression of MMP-2 was in cancer cells: 76.5% (sm), 16.7% (pm), in cancer interstitial region: 92.2% (sm), 58.3% (pm), in transitional mucosal cells: 74.3% (sm), 17.1% (pm) and in transitional mucosal interstitial region: 80.0% (sm), 31.4% (pm). The expression of TIMP-2 was in cancer cells: 41.2% (sm), 16.7% (pm), in cancer interstitial region: 82.4% (sm), 69.4% (pm), in transitional mucosal cells: 57.1% (sm), 22.9% (pm) and in transitional mucosal interstitial region: 89.3% (sm), 57.1% (pm). These results suggest that the expression of MMP-2 and TIMP-2 is induced in the sm cancer stage and is closely related to interactions in the cancer interstitial region for invasion and metastasis in patients with colorectal adenocarcinoma.

A novel weekday-on/weekend-off UFT schedule.

In a step toward a clinical trial, the tumor response and survival of a weekday-on/weekend-off schedule of UFT was compared with its conventional daily schedule in a cancer-bearing rat model. The dose-intensive schedule--600 mg of UFT for 5 days followed by 2 drug-free days--amounts to a weekly dose similar to the conventional schedule of 400 mg/day. The weekday-on/weekend-off schedule provided increased survival and significantly greater antitumor activity than the conventional daily schedule, with no difference in adverse reactions. A study was also conducted in human subjects to measure fluorouracil (5-FU) concentrations that identified the pharmacokinetic activity during the 2 drug-free days of the weekday-on/weekend-off schedule. The plasma 5-FU concentration declined markedly after 24 hours, but the concentration in the tumor remained at a relatively high level after 2 days off the drug. A one-year clinical study evaluated the compliance and toxicity of the weekday-on/weekend-off UFT schedule as adjuvant chemotherapy for colorectal cancer. Based on the findings of all these studies, the weekday-on/weekend-off schedule for UFT as adjuvant chemotherapy for colorectal cancer can be recommended for a clinical trial.

Hypolumbarlordosis: a predisposing factor for preeclampsia.

OBJECTIVE: The aim of this research was to study the relationship between lumbar lordosis and preeclampsia. STUDY DESIGN: We studied lumbar lordosis of 52 primipara patients with pregnancy-induced hypertension and 59 normal primipara pregnant women. We determined the lumbar lordotic deviation (LLD) from lumbar lordosis measurement device in the lateral recumbent position and standing position. Roll-over test was performed for both groups. Resistance index (RI) of internal iliac artery was also measured by color Doppler sonography. RESULTS: The LLD was 33 +/- 6.1 degrees week 20-29 and 34 +/- 6.9 degrees week 30-40, respectively. In patients with preeclampsia, the LLD was 19.8 +/- 6.3 degrees week 20-29, 21.3 +/- 7.9 degrees week 30-40. The LLD in patients with preeclampsia was significantly lower than in the normal pregnant women. The LLD correlated to the increase diastolic pressure after roll-over test significantly. Hypolumbarlordosis was frequently associated with high resistance of peripheral vessel. CONCLUSIONS: We conclude that hypolumbarlordosis is frequently associated with preeclampsia. It is also closely related to the mechanism for positive roll-over test and increase of resistance of vessels. As lumbar lordosis does not change greatly in individuals before pregnancy, hypolumbarlordosis may be a predisposing factor for preeclampsia.

Urinary trypsin inhibitor: a new drug to treat preterm labor: a comparative study with ritodrine.

UNLABELLED: Prevention of preterm delivery is one of the difficult problems facing obstetricians. beta Adrenergic agonists, especially ritodrine, are commonly used in these cases. OBJECTIVES: The aim of this research was to study and compare the effect of urinary trypsin inhibitor (UTI) which has anti-inflammatory anti-cytokine effects with ritodrine in treating preterm labor. STUDY DESIGN: Patients in preterm delivery were randomly selected to be treated either by ritodrine or UTI. In the ritodrine group, uterine contractions were initially suppressed by high doses of ritodrine (up to 300 micrograms/min) and then a maintenance dose was given until 35 weeks of gestation. In the UTI group one vaginal suppository (5000 U) was used daily for 2 weeks. Patients with recurrent preterm uterine contraction during the initial 14 days of treatment, who needed course of other drugs to suppress the contractions, were excluded from the study. Patients responding to the drugs were followed until delivery. Tocolytic index and elastase concentration in the cervical mucus was calculated. Recurrence rate of uterine contraction and time of elongation of pregnancy since the beginning of treatment was calculated. RESULTS: UTI was more effective than ritodrine in inhibition of recurrent uterine contraction and elongation of pregnancy. No side effects could be observed after treatment with UTI for the mother or the fetus. CONCLUSION: UTI may be a new therapeutic method for the inhibition of preterm delivery through suppression of cytokines and inflammatory mediators.

A 72-year-old female with adult Still's disease.

We treated a 72-year-old woman with adult Still's disease. The diagnosis was made on the basis of a prolonged, high grade, quotidian fever, polyarthritis, maculopapular skin rashes and exclusion of other possible diseases. A high serum ferritin value was a key factor both in making the diagnosis and in the follow-up. The patient responded to the administration of oral prednisolone at 30 mg/day, which was tapered to 10 mg/day, with no recurrence of symptoms. This disorder can be an important cause of prolonged fever in the elderly as well as in the younger population.

Follicular bronchiolitis associated with rheumatoid arthritis.

Follicular bronchiolitis is a rare disorder, though it has been recognized as a pulmonary involvement of rheumatoid arthritis in recent years. A 57-year-old woman with rheumatoid arthritis was admitted to the hospital with persistent productive cough and breathlessness on exertion. An open lung biopsy was performed to establish a definite pathologic diagnosis of her disease, and she was diagnosed as having follicular bronchiolitis on the basis of the histopathological findings. It is essential to differentiate this disease from other bronchiolar or lymphoproliferative disorders of the lung.

Longevity in space; experiment on the life span of Paramecium cell clone in space.

Life span is the most interesting and also the most important biologically relevant time to be investigated on the space station. As a model experiment, we proposed an investigation to assess the life span of clone generation of the ciliate Paramecium. In space, clone generation will be artificially started by conjugation or autogamy, and the life span of the cell populations in different gravitational fields (microgravity and onboard 1 x g control) will be precisely assessed in terms of fission age as compared with the clock time. In order to perform the space experiment including long-lasting culture and continuous measurement of cell division, we tested the methods of cell culture and of cell-density measurement, which will be available in closed environments under microgravity. The basic design of experimental hardware and a preliminary result of the cultivation procedure are described.