Biological properties of the native and synthetic lipid A of Porphyromonas gingivalis lipopolysaccharide.

INTRODUCTION AND METHODS: A pentaacyl and diphosphoryl lipid A molecule found in the lipid A isolated from Porphyromonas gingivalis lipopolysaccharide (LPS) was chemically synthesized, and its characteristics were evaluated to reconfirm its interesting bioactivities including low endotoxicity and activity against LPS-unresponsive C3H/HeJ mouse cells. RESULTS: The synthesized P. gingivalis lipid A (synthetic Pg-LA) exhibited strong activities almost equivalent to those of Escherichia coli-type synthetic lipid A (compound 506) in all assays on LPS-responsive mice, and cells. LPS and native lipid A of P. gingivalis displayed overall endotoxic activities, but its potency was reduced in comparison to the synthetic analogs. In the assays using C3H/HeJ mouse cells, the LPS and native lipid A significantly stimulated splenocytes to cause mitosis, and peritoneal macrophages to induce tumor necrosis factor-alpha and interleukin-6 production. However, synthetic Pg-LA and compound 506 showed no activity on the LPS-unresponsive cells. Inhibition assays using some inhibitors including anti-human Toll-like receptor 2 (TLR2) and TLR4/MD-2 complex monoclonal antibodies showed that the biological activity of synthetic Pg-LA was mediated only through the TLR4 signaling pathway, which might act as a receptor for LPS, whereas TLR2, possibly together with CD14, was associated with the signaling cascade for LPS and native lipid A of P. gingivalis, in addition to the TLR4 pathway. CONCLUSION: These results suggested that the moderated and reduced biological activity of P. gingivalis LPS and native lipid A, including their activity on C3H/HeJ mouse cells via the TLR2-mediated pathway, may be mediated by bioactive contaminants or low acylated molecules present in the native preparations having multiple lipid A moieties.

BK virus nephropathy in a patient with ABO-incompatible renal transplantation.

A 43-year-old woman with end-stage renal disease originating from IgA nephropathy entered chronic haemodialysis therapy. She then received an ABO-incompatible living related renal transplantation. Initial immunosuppression consisted of azathioprine, methylprednisolone and tacrolimus. At 155 days after transplantation, the azathioprine was changed to mycophenolate mofetil for continuous graft dysfunction. Furthermore, a total of three courses of anti-rejection therapy was given. At 665 days after transplantation, diagnosis of BK-virus nephropathy was made by immunohistochemical analysis and viral DNA assay. Therefore the immunosuppression therapy was reduced for graft dysfunction. All five renal biopsy specimens were examined retrospectively in order to determine when the BK virus nephropathy had developed. The expressions of SV40 large T antigens were detected from the third (117 days) to the fifth (665 days) biopsies, with increasing numbers of SV40 large T antigen positive cells. In addition, many cells contained inclusion bodies which were already present in the urinary sediment for 3 months post-transplantation. Although it is difficult to make a diagnosis of early stage of BKVN, we have to consider with caution if urinary cells with inclusion body are seen. Awareness of BKVN at the earliest opportunity is important in order to avoid over-immunosuppression.

Tratamento clínco de perfuração esofágica após tratamento endoscópico de divertículo de Zenker: relatro de caso / Clinical treatment of esophageal perforation after endoscopic treatment for Zenker

O divertículo de Zenker é uma alteração da anatomia esofágica que acomete adultos de idade avançada, caracterizadao pela presença de um divertículo posterior, proximal ao músculo cricofaríngeo. De forma precoce, apresenta-se como disfagia transitória e, como o evoluir da doença, como sensação de massa no pescoço e regurgitação. seu tratamento pode ser realizado por endoscopia flexível ou por correção cirúrgica através de miotomia cricofaríngea com diverticulectomia. O objetivo deste trabalhoo é relatar uma possível complicação do tratamento do divertículo de Zenker por endoscopia flexível. Descreve-se o caso de uma paciente feminina, 83 anos, portadora de divertículo de Zenker com início da sintomatologia havia seis meses, submetida a diverticulotomia endoscópica e que algumas horas após o procedimento evoluiu dor e efisema subcutâneo bilateral em região cervical. Esofagografia com contrate iodado evidenciou extravasamento em região cervical. Foi então, instituido manejo conservador, com alta hospitalar assintomática no sexto dia pós-procedimento. Portanto, apesar de considera modalidade de tratamento segura, a diverticulotomia endoscópica não é isenta de riscos

Prevention of secondary osteoporosis postmenopause in hemiplegia.

OBJECTIVES: To study secondary osteoporosis postmenopause in women with hemiplegia and to show the therapeutic effects of etidronate and how osteoporotic conditions relate to the activities of daily living (ADL). DESIGN: Eighty-one postmenopausal women with hemiplegia were admitted within 6 mo of their first cerebrovascular accident. The bone mineral density (BMD) and biochemical markers of bone turnover were measured at the time of admission. Forty women (treatment group) received a 2-wk administration of etidronate. Forty-one women (control group) were not administered etidronate. RESULTS: After completing a 3-mo rehabilitation program, BMD levels were remeasured. ADL was evaluated by FIM. The low ADL group had a larger decrease in BMD than the high ADL group. For the control group, the BMD rate of change on the paretic side of the femoral neck was -9.6%/3 mo for the low ADL group. BMD loss was reduced significantly by the administration of etidronate for the low ADL group. CONCLUSIONS: Results indicate that ADL corresponds to the progression of osteoporosis in postmenopausal women with hemiplegia and that increasing the level of ADL will reduce the progression of osteoporosis. Use of etidronate has also been proven to have a suppressive effect on the BMD decrease in women.

Histomorphometric characteristics and cellular kinetics of colorectal polyps with epithelial serrated proliferation adjacent to carcinoma.

Four cases of colorectal polyps with epithelial serrated proliferation (CP-ESP) with malignant transformation were studied. In CP-ESP adjacent to carcinoma, if the nuclear size in the surface layer was significantly smaller than those in the bottom and the middle layers of the crypts, the specimen was defined as zone formation positive. If there was no significant difference among the layers, the specimen was defined as zone formation negative. Cell kinetics were evaluated using Ki-67 immunostaining. The CP-ESP regions of cases 1 and 2 showed zone formation with inferior and lateral glandular branching, and were qualitatively hyperplastic on cell kinetics. Cases 3 and 4 showed inferior and lateral glandular branching with no zone formation, and were kinetically neoplastic (adenoma). The histogenesis of hyperplastic polyps with atypia (cases 1 and 2) involves the hyperplastic polyp-carcinoma sequence. In contrast, the development of tubulovillous adenoma or serrated adenoma (cases 3 and 4) may involve the tubulovillous adenoma-carcinoma or serrated adenoma-carcinoma sequence.

2'-O,4'-C-ethylene-bridged nucleic acids (ENA) with nuclease-resistance and high affinity for RNA.

Novel nucleosides, 2'-O,4'-C-ethylene nucleosides and their corresponding phosphoramidites, were synthesized as building blocks for antisense oligonucleotides. The 2'-O,4'-C-ethylene linkage of these nucleosides, as well as the linkage of 2'-O,4'-C-methylene nucleosides which are known as bridged nucleic acids (BNA) or locked nucleic acids (LNA), restricts the sugar puckering to the N-conformation. The ethylene-bridged nucleic acids (ENA) showed a high binding affinity for the complementary RNA strand (delta Tm = degrees C/modification) and were approximately 400 and 80 times more nuclease-resistant than natural DNA and BNA/LNA, respectively. These results indicate that ENA have better antisense activity than BNA/LNA.

Loss of left-sided volitional movements caused by a combined lesion of the corpus callosum and right hemisphere:'initiation pseudohemiakinesia'.

We report a right-handed patient who showed a marked loss of unilateral volitional movements of the left limbs after the onset of a cerebral infarction in the combined territories of the right anterior and middle cerebral arteries. The same limbs retained their mobility in acts requiring bilateral sides of the body. This left-sided abnormal behavior resembled motor neglect resulting from lateralized brain damage. Behavioral and neuroradiological findings presented by this patient, however, suggested that callosal disconnection was definitely involved in this symptomatology. We postulate that in this patient, the diseased right hemisphere could no longer initiate movements of the left limbs despite its potential ability to realize them, and that the injured callosum prevented the intact left hemisphere from initiating unilateral voluntary movements of the left limbs. We suggest that this so far undescribed symptomatology be called 'initiation pseudohemiakinesia' in order to be distinguished from other rare forms of unilateral voluntary movements like motor neglect, extinction or directional hypokinesia.

Isolation of the antiulcer compound in essential oil from the leaves of Cryptomeria japonica.

Essential oil from the leaves of Tateyamasugi (Cryptomeria japonica) exhibited strong inhibitory activity on ulceration induced by HCl/ethanol, HCl/aspirin, water-immersion stress and pylorus-ligation. We separated the antiulcer compounds from cedar essential oil by use of distillation and chromatography. As a result, terpinen-4-ol, a monoterpene, and elemol, a sesquiterpene, were isolated as active compounds. The antiulcer activity of the former was more potent than that of the latter. Terpinen-4-ol was a mixture of optical isomers and each possessed potent antiulcer activity. Secretion of gastric juice and output of acid and pepsin activity were lowered by terpinen-4-ol. These results suggest that terpinen-4-ol isolated from cedar essential oil could be a valuable antiulcer agent.

Ultrastructural evaluation of apoptosis induced by Helicobacter pylori infection in human gastric mucosa: novel remarks on lamina propria mucosae.

It has been considered that Helicobacter pylori (H. pylori) infection is a major cause of human gastritis and gastroduodenal ulcers (G-DU). Many investigations of the relationship between H. pylori and apoptosis have been reported recently. However, these studies focused mostly on epithelium, using the TUNEL method. In the present study, we evaluated by electron microscopy the occurrence of apoptosis in the mesenchymal cells of lamina propria mucosae infected with H. pylori. Gastric biopsy specimens from 37 H. pylori-infected G-DU patients and 8 noninfected volunteers were examined with both light and electron microscopy and analyzed by the TUNEL method. The TUNEL method showed no significant difference between H. pylori-infected and noninfected cases. In contrast, electron microscopy revealed significant numbers of apototic fibroblasts and smooth muscle cells in H. pylori-infected lamina propria mucosae, with a diminished number of collagen fibers in surrounding areas. These areas showed edematous changes histopathologically. These results indicated that H. pylori infection induces apoptosis of fibroblasts and smooth muscle cells in lamina propria, with decrease in the numbers of collagen fibers, suggesting that these alterations may be affected by exaggerate acid secretion, decrease mucus protecting factors, and result in ulcer formation.

A case of ABO-incompatible renal transplant patient with non-insulin-dependent diabetes mellitus; long-standing observation of serial glomerular change by protocol biopsy.

A 41-yr-old patient with non-insulin-dependent diabetes mellitus (NIDDM), before and after ABO-incompatible renal transplant, is reviewed using serial protocol biopsy. Although she recovered from delayed hyperacute rejection (DHAR) immediately post-transplantation, her graft function deteriorated gradually. A mild acute transplant glomerulitis, noted at the 155th day post-transplantation, progressed to pronounced chronic transplant glomerulopathy over 5 yr. In the specimen of the last biopsy, at 5 yr post-transplantation, glomeruli demonstrated an exudative hyaline lesion, which was characteristic of diabetic nephropathy in addition to chronic transplant glomerulopathy. Therefore, we made a diagnosis of this glomerular lesion as chronic transplant glomerulopathy complicated by diabetic glomerulopathy. Considering the result of this case, the protocol biopsy is a useful procedure to diagnose an accurate cause of graft dysfunction in individual cases. It is concluded that the protocol biopsy is apparently useful for the detection of various pathological processes occurring in allograft and may contribute to a strategy for improvement of graft survival.

Martefragin A, a novel indole alkaloid isolated from red alga, inhibits lipid peroxidation.

Martefragin A (1), a novel indole alkaloid, was isolated from a red alga, Martensia fragilis, by repeated column chromatography. The structure of 1 was elucidated on the basis of spectral analysis of its methyl ester (2), including 1H- and 13C-NMR, 1H-1H correlation spectroscopy (COSY), and 13C-1H COSY. A single crystal X-ray analysis of the hydrochloride of 1 confirmed the assignment. Martefragin A (1) showed inhibitory activity on NADPH-dependent lipid peroxidation in rat liver microsomes. The IC50 values of 1, alpha-tocopherol and ascorbic acid were 2.8, 87 and 200 microM, respectively.

Prevalence and conditions of urinary incontinence among the elderly.

In Japan, elderly disorders and diseases have markedly increased in recent years, because of rapid aging and an increasing number of older persons. The situation is creating serious social and community problems. These disorders, particularly dysuria and urinary incontinence (UI), disturb the quality of life (QOL) in latelife. Few reports on UI have been published, but precise investigation into the community level remains to be made. Our presentation is the development, implementation and evaluation of elderly UI in Kumamoto Prefecture. This study includes 2,304 people (male: 856, female: 1,448), over 65 years of age, living in two different communities; one is an urban (K) and the other is a typical rural area (S). The rate of UI was in homebound elderly persons, male: 4.7%, female: 11.3%, and in nursing home residents, male: 16.2%, female: 23.2%. The condition of UI was: almost Urinary Urgency in male (61.5%), and Stress Incontinence (such as, caused by coughing, sneezing, and exercise) in female (46.3%). The influence of UI on the activity of daily life was investigated. Most of the male cases were giving concerns for family and community. In contrast, females hesitated to participate in group excursions and outdoor exercise, and had a tendency to live alone or indoors. However, most persons (81.5%) with UI did not visit a physician. From this investigation, we conclude that a community health care program and public support system are essential for proper understanding and solution of the elderly UI problem.

Dietary diosgenin attenuates subacute intestinal inflammation associated with indomethacin in rats.

We investigated the effects of dietary diosgenin (Dio), a plant-derived sapogenin, on indomethacin (Indo)-induced intestinal inflammation and alterations in bile secretion in rats. In anesthetized rats, bile secretion, intestinal inflammation, and blood chemistry were assessed 3 days after two subcutaneous injections of Indo given 24 h apart. Dio (> 80 mg.kg-1.day-1) pretreatment significantly inhibited weight and food intake decreases and intestinal inflammation. This protective effect was confirmed by examination of gross and histological findings and intestinal myeloperoxidase activity. Dio significantly increased biliary cholesterol (Chol) output and prevented the decreases in bile flow, bile acid output, and biliary alpha-muricholic acid and the increases in biliary hyodeoxycholic acid, deoxycholic acid, and hydrophobicity index of bile. Significantly more biliary Chol and phospholipids were present in macromolecules separate from bile acids and Indo in Dio-treated rats. Dio significantly increased the elimination constant of Indo and reduced plasma Indo levels at 3 and 12 h but did not influence biliary secretion of Indo for 3.5 h after injection. Although Dio dose-dependently attenuated subacute intestinal inflammation and normalized bile secretion in this model, it may also compromise the anti-inflammatory action of indo.

Interleukin-6 is a useful marker for early prediction of the severity of acute pancreatitis.

Twelve patients with acute pancreatitis admitted to our department between January 1993 and December 1994 were studied prospectively and classified into two groups (severe group, five patients; mild group, seven patients), according to the criteria for grading severity of acute pancreatitis proposed by the Research Committee for Intractable Diseases of the Pancreas, Japanese Ministry of Health and Welfare (1990). To evaluate markers for early estimation of the severity of acute pancreatitis, we measured serum changes in various parameters. In the severe group interleukin-6 (IL-6) levels were increased significantly 5, 24, 72, and 120 h after the onset (p < 0.01), compared with the mild group. C-reactive protein (CRP), thrombin antithrombin III, and alpha 2-plasmin inhibitor plasmin complex levels were significantly increased only at the 72-h time point. Peak values of interleukin-8 (IL-8) and soluble human E selectin were observed at 5 and 72 h, respectively, after the onset. There was a significant correlation between IL-6 at 5 h and both pancreatic secretory trypsin inhibitor (r = 0.85) and CRP (r = 0.94) at 72 h. We therefore conclude that IL-6 is a useful marker for assessment of the severity of acute pancreatitis in its early stages.

[Production of medicinal plants by soilless culture system. I. Studies of morphological characteristics and saikosaponins content in Bupleurum falcatum cultivated by Ebb & Flood system].

We studied that the morphological and histological characteristics, and the content (%DW) of saikosaponins on the root of Bupleurum falcatum cultivated in an Ebb & Flood system (E & F), a kind of soilless culture system, by both the direct sowing and the transplanting methods, and that effects of pinching on the root growth and the content (%DW) of saikosaponins in each part of root. Yield of root and content (%DW) of saikosaponins in each part of root, 8-months-old, cultivated in E & F by both methods were at the same level as that cultivated for the same period in soil condition by generally standard procedures. Morphological characteristics of the root cultivated by the direct sowing method were the same appearance as that by soil condition, but by the transplanting method main root branched off in all direction and the lateral root were more developed than by the direct sowing method. By pinching lignification in xylem on the main root were inhibited, but the dried weight of total root part and content (%DW) of saikosaponins in each part of the root were not shown to be significantly changed.

Duodenal carcinoid tumor: endosonographic imaging and endoscopic resection.

A 75-yr-old Japanese woman was shown by endoscopy to have a sessile polyp in the duodenal bulb. Endoscopic biopsy confirmed the diagnosis of carcinoid tumor. The endoscopic ultrasound probe showed a 6-mm hypoechoic tumor that was confined to the submucosal layer; the underlying muscularis propria layer was intact. On the basis of these findings, endoscopic resection was chosen, and the tumor was completely resected by the strip biopsy technique with prior hypertonic saline and epinephrine injection; there were no complications.

Circulatory and plasma catecholamine responses to mental stress in young subjects with two different types of hypertension.

To determine whether any differences exist between young male subjects with elevated diastolic and systolic blood pressure (BP) and those with only an elevated systolic BP, the responses of BP and plasma catecholamines to a mental arithmetic test were studied in 11 young men (mean age of nineteen years) with BP of > or = 140/90 mm Hg at a routine health check-up (group I-hypertension [HT]), 26 age-matched men with only elevated systolic BP (> or = 140 mm Hg) (group II-HT), and 12 age-matched normotensive (NT) men (< 140/90 mm Hg). During an arithmetic test, group I-HT showed a significantly higher increment of systolic BP (+14.5-18.0%) than group II-HT (+9.3-10.2%) and NT controls (+6.4-8.2%). However, no significant difference in BP response was seen between group II-HT and NT controls. Plasma norepinephrine in group I-HT showed a significant increase after the test, (171 +/- 23-->202 +/- 27 pg/mL), whereas group II-HT and NT controls showed no change. No significant difference occurred in epinephrine response among the three groups. These results suggest that the young male subjects with screening BP above 140/90 mm Hg are hyperreactive to a mental stress as compared with subjects with only elevated systolic BP or NT controls.

Increased urinary dopamine excretion in young patients with essential hypertension.

The evidence that some older patients with essential hypertension have low urinary dopamine excretion has brought into question the levels of urinary dopamine and plasma dopa, the major source of urinary dopamine, in young patients with essential hypertension. Twenty-four-hour urine sodium, creatinine, dopamine and noradrenaline and plasma dopa were evaluated in 48 patients with essential hypertension aged 18 to 27 years and 25 normotensive subjects. In comparison with age-matched normotensive subjects, the hypertensive patients had higher urinary dopamine (1920 +/- 80 vs 1520 +/- 130 nmol/day, p < 0.01) and noradrenaline (216 +/- 11 vs 179 +/- 12 nmol/day, p < 0.05) excretion. There was a significant correlation between urinary dopamine and noradrenaline excretion. There was no difference in plasma dopa levels between normotensive and hypertensive subjects. These results suggest that the elevated conversion of dopa to dopamine in the kidney is leading to increased urinary dopamine excretion in young patients with essential hypertension.

Leisure time physical activity and insulin resistance in young obese students with hypertension.

To investigate the hypothesis that insulin resistance plays a role in the etiology of hypertension and hyperlipidemia, we measured serum lipid levels, the fasting glucose/insulin ratio, and the insulin response to oral glucose (GTT) in a group of young obese subjects (n = 21) with hypertension and normal glucose tolerance and in normotensive subjects (n = 36) with normal glucose tolerance, matched for age and body mass index. Leisure time physical activity was evaluated by a questionnaire outlining three levels of physical activities during leisure time. Subjects with hypertension had higher fasting serum insulin (19 +/- 2 v 13 +/- 1 microU/mL, P < .01) and lower fasting glucose/insulin ratio (5.3 +/- 0.2 v 7.1 +/- 0.5 mg/dL/microU/mL, P < .01) than normotensive subjects. Subjects with hypertension had higher peak serum insulin and lower plasma glucose area/insulin area ratio in response to glucose (1.8 +/- 0.2 v 2.4 +/- 0.2 mg/dL/microU/mL, P < .05) than normotensive subjects. Serum total cholesterol, low-density cholesterol, and triglycerides were higher in the obese hypertensive subjects than in obese normotensive ones. Blood pressure correlated with either fasting serum insulin, fasting glucose/insulin ratio, or glucose area/insulin area ratio during GTT. The level of leisure time physical activities was lower in obese hypertensive subjects than in obese normotensive ones. There were significant correlations between the levels of physical activity and the fasting plasma glucose/insulin ratio (r = 0.371, P < .01) or the fasting serum insulin concentration (r = -0.282, P < .05). The study provided evidence that a low level of leisure time physical activity is associated with insulin resistance and resultant hyperinsulinemia, which are the key metabolic abnormalities that link hypertension, obesity, and hyperlipidemia in young subjects.