A small-molecule compound targeting CCR5 and CXCR3 prevents airway hyperresponsiveness and inflammation.

Asthma is associated with increased numbers of T-cells in the lung. CC chemokine receptor (CCR)5 and CXC chemokine receptor (CXCR)3 have been reported to play important roles in the lung T-cell homing pathway, and may be potential targets for asthma therapy. The aim of the present study was to investigate the role of CCR5 and CXCR3 in allergen-induced acute asthma and to determine whether a novel small-molecule compound, TAK-779, targeting CCR5 and CXCR3 can attenuate allergic airway responses. Mice were sensitised with ovalbumin (OVA). mRNA expression of chemokine receptors in the lung were measured after the challenge with either aerosolised phosphate-buffered saline or OVA. OVA-sensitised mice were also treated with TAK-779. Respiratory function was measured, bronchoalveolar lavage was performed, and blood and lung samples were obtained. OVA challenge increased CCR3, CCR5 and CXCR3 expression in the lung. Treatment with TAK-779 significantly attenuated altered respiratory function and pulmonary allergic inflammation. The beneficial effect was associated with reduced expression of CCR5 and CXCR3 in the lung. These data demonstrate that blockade of CC chemokine receptor 5 and CXC chemokine receptor 3 using TAK-779, a synthetic nonpeptide compound, can prevent the development of asthma features in a mouse model. Thus, CC chemokine receptor 5 and CXC chemokine receptor 3 may be potential targets for asthma therapy.

Purification and characterization of cytochromes P-450 from liver microsomes of 3-methylcholanthrene-treated crab-eating monkeys.

Two forms of cytochrome P-450 (P-450MC1 and P-450MC2) were purified from liver microsomes of crab-eating monkeys (Macaca irus) treated with 3-methylcholanthrene (MC). Monkey P-450MC1 preparation had a specific content of 14.0 nmol/mg protein and showed a main protein band with a minimum molecular weight of 52,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Monkey P-450MC2 preparation had a specific content of 12.1 nmol/mg protein and a minimum molecular weight of 54,000. The carbon monoxide-reduced difference spectral peaks of monkey P-450MC1 and P-450MC2 were at 448 and 447 nm, respectively. In the reconstituted system, monkey P-450MC2 had high activities for benzo[a]pyrene 3-hydroxylation and 7-ethoxycoumarin O-deethylation. Monkey P-450MC1 had low activities toward these two substrates and a high activity for benzphetamine N-demethylation. Monkey P-450MC1 and P-450MC2 were detected by immunoblotting using an antibody prepared against rat cytochrome P-450c, which is a major form of cytochrome P-450 in liver microsomes of MC-treated rats. These results suggested that the molecular properties of cytochrome P-450 in liver microsomes of crab-eating monkeys treated with MC are similar to those in rats.

Effect of endothelin-1(1-31) on p38 mitogen-activated protein kinase in cultured human mesangial cells.

It was reported that human chymase cleaves big endothelins (ETs) at the Tyr31-Gly32 bond and produces 31-amino acid ETs(1-31). In this study, we investigated the effect of ET-1(1-31) on p38 mitogen-activated protein kinase (p38-MAPK) activity in human mesangial cells (HMCs). By measuring the kinase activity, we demonstrated that ET-1 (1-31) activated the p38-MAPK dose-dependently (10(-9) M to 10(-7) M), which was inhibited by SB203580. The p38-MAPK activation induced by ET-1(1-31) peaked at 10 minutes. BQ123 almost abolished ET-1(1-31)-induced p38-MAPK activation, whereas BQ788 failed to inhibit it. These findings suggest that the stimulatory effect of ET-1(1-31) on p38-MAPK activation is mediated through ET(A) or ET(A)-like receptor. In conclusion, ET-1(1-31) induced increase in p38-MAPK activation in cultured HMCs.

A new technique for variable-load cholangiomanometry. Implication for biliary tract surgery.

BACKGROUND/AIMS: Variable-load cholangiomanometry was performed to obtain data on terminal biliary function during the surgical treatment of cholelithiasis. The decision of whether or not to perform a definitive biliary drainage procedure was based on the results of this test. MATERIAL AND METHODS: The rate of perfusion was reduced in four steps from 15.3 ml/min to 1.2 ml/min, and the resultant perfusion pressures were plotted. The gradient produced by the straight line was considered the resistance, R. The baseline pressure without perfusion was regarded as the static pressure, P. RESULTS: A review of 444 patients with cholelithiasis who underwent intraoperative cholangiomanometry during the past 11 years led to the following indications for definitive biliary drainage: (1) R > 10 units and P > 200 mm H2O, (2) if only R or P is elevated, priority is given to R, and (3) if the elevation of R or P is borderline, the presence of a type I curvature in the segment of low flow rate is an indication for surgery. CONCLUSION: By performing an intraoperative cholangiomanometry concrete indications for a biliary drainage procedure can be defined.

Assessment of postoperative digestive function after pancreatico-duodenectomy: a comparison of reconstruction techniques.

BACKGROUND/AIMS: Malabsorption is a frequent complication following pancreatico-duodenectomy (PD) for pancreatic carcinoma. Gastrojejunostomy followed by Billroth I type of reconstruction (PD III) has been advanced to prevent this disorder. We compared postoperative digestion and absorption determined by 131I-triolein, D-xylose and pancreatic function diagnostant (PFD) after extensive PD followed by one of two reconstructive procedures. MATERIAL AND METHODS: Postoperative digestive and absorptive functions in patients operated by Child's method (PD II) were compared with patients in whom the stomach emptied proximal to the pancreas and bile duct (PD III). RESULTS: The absorption of D-xylose was significantly higher (1.24 +/- 0.36 g vs. 0.72 +/- 0.21 g) in the PD III group. No difference was noted in pancreatic endocrine secretion between the two groups. Biliary scintigraphy revealed increased bile secretion in the PD III group. CONCLUSIONS: These results suggest that PD III is superior to PD II following extensive PD.

Endometriosis in cynomolgus monkeys retired from breeding.

Twenty-seven cases of endometriosis were found in 94 female cynomolgus monkeys retired from breeding. The ages of the affected monkeys (28.7% of all the monkeys) ranged from 11 years to 23 years of age with an average of 15.3 years. There was no significant difference in the incidence of endometriosis between groups which had or had not undergone a cesarean section. Time after the last pregnancy was significantly longer in the affected monkeys than in the non-affected ones. These cases were histopathologically classified into four types: 1) pelvic endometriosis with formation of endometrial cysts (51.8% of the affected monkeys), 2) pelvic endometriosis invading other organs without formation of endometrial cysts (3.7%), 3) uterine adenomyosis (22.2%), 4) combined pelvic endometriosis and uterine adenomyosis (22.2%).

Efficacy of systemic corticosteroid treatment for anosmia with nasal and paranasal sinus disease.

Systemic administration of corticosteroids was attempted in the treatment of olfactory loss resistant to topical corticosteroid treatment in patients with nasal and paranasal disease and post-upper respiratory infection. Significant efficacy was achieved with a short course of high-dose oral corticosteroids in patients with non-allergic sinus disease. On the other hand, anosmia induced by upper respiratory infection failed to respond to systemic corticosteroid treatment, suggesting permanent damage to the olfactory receptor cell. The underlying mechanism of effectiveness observed in patients with sinus disease may be explained by improvement of the mucosal thickening of the olfactory fissure, leading to the access of an odorant to the olfactory neuroepithelium.

Concentrations of C-reactive protein in normal monkeys (Macaca irus) and in monkeys inoculated with Bordetella bronchiseptica R-5 and measles virus.

The concentrations of C-reactive protein (CRP) in serum from normal crab-eating monkeys (Macaca irus) were measured by means of a monkey-specific turbidimetric immunoassay (TIA), and the changes in the serum CRP concentrations in crab-eating monkeys inoculated with Bordetella bronchiseptica R-5 and measles virus (Ichinose or NK 3 strain) were also examined. The CRP concentrations in sera from 54 normal crab-eating monkeys ranged from 0 to 8.3 microg/ml (mean 2.2 +/- 1.9). No significant difference was found in the CRP concentrations between males and females (p > 0.05). The concentrations of CRP in the sera from four crab-eating monkeys inoculated intrabronchially with 10(9) live B. bronchiseptica increased gradually to a peak at 2 days after inoculation. The peak concentrations of CRP were from 102.4 to 313.2 microg/ml, 54-96 times the preinoculative values of 1.9-5.6 microg/ml. When the same four crab-eating monkeys were inoculated intrabronchially with measles virus 34 days after inoculation of B. bronchiseptica, the serum CRP concentrations did not increase. Monitoring of CRP is useful for assessing monkeys with acute B. bronchiseptica infection and will probably be of value in the diagnosis of other bacterial infections.

[Progress of preoperative diagnosis of pancreatic tumor--with special reference to extension of pancreatic cancer].

Preoperative diagnosis of the extension of pancreatic cancer is very important for determining the range of resection. We evaluated the preoperative imaging diagnosis of the tumors and compared it with the pathological diagnosis. Out of 68 pancreatic head cancers, the percentage of correct diagnosis of serosal invasion was 69.1%, that of over-estimate was 23.5% and that of under-estimate was 7.4%. In the case of retroperitoneal invasion, a correct diagnosis had been made in 79.1%, over-estimate in 4.5% and under-estimate in 16.4%. In the case of portal vein invasion, these percentages were 52.8%, 39.6% and 7.5%, respectively. In the case of lymph node involvement, these percentages were 85.3%, 2.9% and 11.8%, respectively. The percentage of preoperative over-estimate was higher for serosal invasion and that of preoperative under-estimate was higher for retroperitoneal invasion. In the case of portal vein invasion, the percentage of preoperative over-estimate was relatively high. Of the 12 patients who had paraortic lymph node metastases, 6 patients had a single metastatic node in the paraortic area. The minimum size of these nodes was 2 x 2 mm. Only 5 cases could be diagnosed preoperatively. We evaluated the usefulness of abdominal CT scan and superior mesenteric arteriography for the diagnosis of superior mesenteric arterial wall invasion. In both methods, the over-estimate rate was 0%, but the under-estimate rate was relatively high. Conclusively the preoperative diagnosis of the extension of the pancreatic cancer by conventional imaging methods has some limits.

[Usefulness of power Doppler sonography for the assessment of subpleural lesions].

We investigated the usefulness of color Doppler power mode imaging for the assessment of subpleural lesions in 48 patients (27 with pneumonia, 4 with pulmonary abscesses, 12 with primary lung cancer, and 5 with metastatic lung cancer). We classified the patterns obtained by color flow imaging of subpleural lesions into six groups: type 0, no color flow; type I, spotty color flow; type II, linear color flow; type III, branchy color flow; and type IV, tortuous color flow, with type IV-A, for partial tortuous flow and type IV-B, for general tortuous flow. The color Doppler power mode proved better than velocity mode in terms of ability to generate clear color flow patterns. Color flow patterns obtained in power mode on the patients with pneumonia differed significantly from the patterns obtained on the lung cancer patients. Although the color flow patterns observed in power mode differed significantly for the benign and malignant groups, no statistically significant differences were observed in velocity mode. These findings illustrated the usefulness of color Doppler power mode imaging as a means of diagnosing benign and malignant subpleural lesions.

[Dynamic behavior of guinea pig middle ear].

The dynamic behavior of the middle ear was studied in living guinea pigs by using a laser Doppler vibrometer coupled to a compound microscope. Placing glass microbeads 20 microns in diameter on four points of the tympanic membrane and six points of the ossicles, their velocity amplitudes were measured under a constant stimulus of 65 dB SPL. Velocity responses of the four points on the tympanic membrane differed over the frequency range from 0.1 kHz to 3 kHz. At low frequencies, the malleus and incus rotated around an axis running from the malleus head to the incus short process. At middle frequencies, the whole ossicle had a piston-like motion. At high frequencies, the malleus and incus rotated around a vertical axis running through the ossicles, while the stapes had a hinge-like movement.

[Multiple primary cancers of oral cavity and pharynx].

Multiple primary cancers were found in 23 of 68 patients (34%) with an index cancer in the oral cavity or pharyngeal area treated in our institute from June 1995 to July 1998. Four cases had triple primary cancers. All 68 cases underwent upper and lower gastrointestinal endoscopy as well as ultrasonography of the liver. Lung CT was performed in cases with abnormal findings on chest roentgenograms. Multiple cancers were found in 5 of 25 oral cavity cases (20%), 6 of 14 mesopharynx cases (43%) and 12 of 29 hypopharynx cases (41%). Nine of 23 cases (39%) were synchronous and 14 (61%) were metachronous. Eighteen of 27 (69%) secondary cancers occurred in the upper aerodigestive tract with an especially high incidence (22%) in the esophagus. Gastroendoscopy also revealed 7 neoplastic lesions, aside from cancers, with the total abnormal rate of 24% (24/68). Thus, gastroendoscopy is useful for the diagnosis of multiple primary cancers. The frequency of multiple primary cancers in males (33%) was not different from that in females (35%). The average age of multiple primary cancer patients (65.1 years) was a little higher than that of single cancer patient (62.7 years). Smoking or drinking was not related to the incidence of multiple cancers. The interval between the first and the second cancer in metachronous cancer cases was 25.5 months on average, and within 4 years in 71% (10/14) of the cases. This result suggests that close follow-up including endoscopy should be required for at least 4 years after treatment of oral or pharyngeal cancer. Radical treatment for each of the multiple cancer lesions was performed in 22 of 23 cases, and the mortality rate of multiple primary cancer cases was not significantly different from that of single cancer cases. Among 7 cases who died of disease, 5 cases died of distant metastasis, suggesting that control of distant metastasis is an important issue in the treatment of multiple primary cancers.

[Canonical discriminant analysis for pregnancy-related changes in hematological and serum biochemical values in tamarins (Saguinus spp].

Hematological and biochemical values obtained from 9 monkeys (Saguinus labiatus and S. mistax) during pre- and postpartum periods were analyzed by canonical discriminant analysis (discriminant analysis with reduction of dimensionality). All animals used were of wild origin and had been maintained under uniform environmental conditions at N. I. H., Japan. The items examined were as follows: white blood cell count (WBC), red blood cell count (RBC), hematocrit value (Ht), hemoglobin concentration (Hb), total protein concentration (TP), blood urea nitrogen (BUN), albumin concentration (ALB), albumin-globulin ratio (A/G), glutamic oxaloacetic transaminase activity (GGT), glutamic pyruvic transaminase activity (GPT), alkaline phosphatase activity (ALP) and total cholesterol concentration (CHO). The data obtained in the pre- and postpartum periods were divided into six chronological groups. The prepartum period was divided into Group I: weeks 15-10; Group II: weeks 9-7; Group III: weeks 6-4; and Group IV: weeks 3-0. The postpartum period was divided into Group V: weeks 0-4 and Group VI: weeks 5-7). In the later pregnancy period (Groups III and IV), significant decreases in RBC, Ht, Hb, TP and ALB, and a significant increase in CHO were observed. These values in the blood and serum continued after delivery (Groups V and VI). Results of canonical discriminant analysis showed that the value of the first canonical variate decreased according to the progress of pregnancy. The postpartum groups showed negative values. Although groups in the early

Weight and size of the placenta in cynomolgus monkeys (Macaca fascicularis).

A gross anatomical survey of the placental development was carried out in 24 cynomolgus monkeys (Macaca fascicularis) of 4, 5, 6, 7, 8, 9, 10, 15 and 20 weeks of gestational age. Eight of the 24 placentas examined were found to be single discoidal and the remaining 16 bidiscoidal. The weight and size of placentas of each animal were measured. The average weight of the 20th week placentas was about 94 times heavier than that of the 4th week ones; the average diameter of the former was about 5 times larger than that of the latter.

Detection of macaque chorionic gonadotropin in the placenta of the cynomolgus monkey (Macaca Fascicularis).

The biologically detectable period of macaque chorionic gonadotropin (mCG) was estimated by the mouse uterine weight method with 22 placentas of the cynomolgus monkey at various pregnancy stages. The hormone was detectable in the placental extracts of the 4th and 5th week of gestation, but not in those of the 6th week or later. This fact may be compatible with our previous finding that mCG was not detectable in urine after the 6th week of gestation in the cynomolgus monkey.

Quercetin inhibits Shc- and phosphatidylinositol 3-kinase-mediated c-Jun N-terminal kinase activation by angiotensin II in cultured rat aortic smooth muscle cells.

Angiotensin II (Ang II) induces vascular smooth muscle cell (VSMC) hypertrophy, which results in various cardiovascular diseases. Ang II-induced cellular events have been implicated, in part, in the activation of mitogen-activated protein (MAP) kinases. Although it has been proposed that daily intake of bioflavonoids belonging to polyphenols reduces the incidence of ischemic heart diseases (known as "French paradox"), the precise mechanisms of efficacy have not been elucidated. Thus, we hypothesized that bioflavonoids may affect Ang II-induced MAP kinase activation in cultured rat aortic smooth muscle cells (RASMC). Our findings showed that Ang II stimulated rapid and significant activation of extracellular signal-regulated kinase (ERK) 1/2, c-Jun N-terminal kinase (JNK), and p38 in RASMC. Ang II-induced JNK activation was inhibited by 3,3',4',5,7-pentahydroxyflavone (quercetin), a major bioflavonoid in foods of plant origin, whereas ERK1/2 and p38 activation by Ang II were not affected by quercetin. Ang II caused a rapid tyrosine phosphorylation of Src homology and collagen (Shc), which was inhibited by quercetin. Quercetin also inhibited Ang II-induced Shc.p85 association and subsequent activation of phosphatidylinositol 3-kinase (PI3-K)/Akt pathway in RASMC. Furthermore, LY294002, a PI3-K inhibitor and a quercetin derivative, inhibited Ang II-induced JNK activation as well as Akt phosphorylation. Finally, Ang II-induced [(3)H]leucine incorporation was abolished by both quercetin and LY294002. These findings suggest that the preventing effect of quercetin on Ang II-induced VSMC hypertrophy are attributable, in part, to its inhibitory effect on Shc- and PI3-K-dependent JNK activation in VSMC. Thus, inhibition of JNK by quercetin may imply its usefulness for the treatment of cardiovascular diseases relevant to VSMC growth.

Three acylated saponins and a related compound from Pithecellobium dulce.

Four new oleanane-type triterpene glycosides, pithedulosides H-K (1-4), were isolated from the seeds of Pithecellobium dulce. Their structures were established by extensive NMR experiments and chemical methods. Compounds 1-3 comprised acacic acid as the aglycon and either monoterpene carboxylic acid and its xyloside or monoterpene carboxylic acid as the acyl moiety at C-21. The oligosaccharide moieties linked to C-3 and C-28 were determined as alpha-L-arabinopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-(1 -->6)- [beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl and alpha-L-arabinofuranosyl-(1-->4)-[beta-D-glucopyranosyl-(1-->3)]-alpha- rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl ester, respectively. Compound 4 was established as an echinocystic acid 3-O-glycoside having the same sugar sequences as 1-3. Also obtained in this investigation was the known compound 5, which was identified as echinocystic acid 3-O-beta-D-xylopyranosyl- (1-->2)-alpha-L-arabinopyranosyl-(1-->6)-[beta-D-glucopyranosyl-(1 -->2)]- beta-D-glucopyranoside.