Causal involvement of mammalian-type cryptochrome in the circadian cuticle deposition rhythm in the bean bug Riptortus pedestris.

Mammalian-type CRYPTOCHROME (CRY-m) is considered to be a core repressive component of the circadian clock in various insect species. However, this role is based only on the molecular function of CRY-m in cultured cells and it therefore remains unknown whether CRY-m is indispensable for governing physiological rhythms at the organismal level. In the present study, we show that RNA interference (RNAi) targeting of cry-m in the bean bug Riptortus pedestris disrupts the circadian clock governing the cuticle deposition rhythm and results in the generation of a single cuticle layer. Furthermore, period expression was induced in cry-m RNAi insects. These results verified that CRY-m functions as a negative regulator in the circadian clock that generates physiological rhythm at the organismal level.

Helicobacter pylori infection enhances N-methyl-N-nitrosourea-induced stomach carcinogenesis in the Mongolian gerbil.

No previous report has demonstrated the relationship between Helicobacter pylori (HP) infection and gastric carcinogenesis in an experimental animal model. A total of 170 male Mongolian gerbils (MGs) were divided into nine groups (18 < or = n < or = 20 for each group). MGs of four groups were inoculated with HP before or after continuous N-methyl-N-nitrosourea (MNU) administration via the drinking water. Both intestinal-type and diffuse-type adenocarcinomas, including signet ring-cell carcinomas, were found at 40 weeks after the study commenced, but only in the HP inoculation groups with MNU exposure and not in the MNU alone or HP inoculation alone control groups. The present findings demonstrate that HP infection increases the incidence of MNU-induced adenocarcinoma of the glandular stomach in MGs.

Timing of N-methyl-N-nitrosourea administration affects gastric carcinogenesis in Mongolian gerbils infected with Helicobacter pylori.

In Mongolian gerbils, the gastric mucosa shows dramatic changes after Helicobacter pylori inoculation. The influence of the timing of N-methyl-N-nitrosourea (MNU) administration after H. pylori inoculation on gastric carcinogenesis was investigated. Ninety-two gerbils were divided into four groups. One group was given MNU at 24 weeks, and another at 1 week, after H. pylori inoculation. The other groups received only MNU. Stomachs were excised for histological examination 20 weeks after completing the MNU treatment. Carcinomas arose only in animals given MNU 1 week after H. pylori inoculation. In the relationship between H. pylori and gastric carcinogenesis, H. pylori is a promoter rather than an initiator, and other environmental factors (the timing of MNU administration) play important roles.

Purification and characterization of a trypsin-like protease in the submandibular gland of rats.

A trypsin-like protease (named RSP-V) was purified to homogeneity from rat submandibular glands by isoelectric focusing and high-performance liquid chromatography. The purified enzyme had an isoelectric point of 5.3 and an apparent molecular weight of 25,000, and consisted of two subunits with molecular weights of 19,500 and 6,000. RSP-V hydrolyzed BAEE, BAPA, and TAME, but not ATEE or BTPA. It had an optimum pH at around 10.0. RSP-V was strongly inhibited by soybean trypsin inhibitor, aprotinin, leupeptin, antipain, and benzamidine, but not by ovomucoid trypsin inhibitor, p-CMB, or iodoacetic acid. This enzyme partly resembled, but was not identical with, tonin. It was also different from kallikrein, salivain, and glandulain in rat submandibular gland. Although the physiological role of RSP-V has not yet been clarified, this enzyme inactivated dopa decarboxylase alone among catecholamine-synthesizing enzymes.

Optically active beta-ketoiminato cationic cobalt(III) complexes: efficient catalysts for enantioselective carbonyl-ene reaction of glyoxal derivatives.

[see reaction]. Optically active beta-ketoiminato cationic cobalt(III) complexes were synthesized as effective Lewis acid catalysts for the enantioselective carbonyl-ene reaction. In the presence of a catalytic amount of cobalt(III) hexafluoroantimonate derived from the optically active 1,2-diphenyl-1,2-ethanediamine, the carbonyl-ene reaction with a variety of alkenes and glyoxal derivatives smoothly proceeded to afford the corresponding homoallylic alcohols in good-to-high yields with high enantioselectivities.

Reductive desymmetrization of 2-alkyl-1,3-diketones catalyzed by optically active beta-ketoiminato cobalt complexes.

[reaction: see text] The reductive desymmetrization of acyclic 1,3-diketones was achieved for the first time by catalytic borohydride reduction in the presence of optically active beta-ketoiminato cobalt(II) complex catalysts. In this reaction, various 2-substituted-1,3-diaryl-1,3-propanediones were converted into the corresponding optically active 2-substituted-1,3-diaryl-3-hydroxypropanone in good-to-high yields with excellent diastereo- and enantioselectivities and high catalytic efficiencies.

Highly chemo-, diastereo-, and enantioselective reduction of 1,2-dialkyl-3-aryl-1,3-diketones for preparation of aldol-type compounds.

[reaction: see text]. Highly chemo-, diastereo-, and enantioselective borohydride reduction of 2-substituted-1,3-diketones was achieved in the presence of the optically active beta-ketoiminato cobalt complex catalysts to afford the optically active 2-substituted-3-hydroxyketones. The present catalytic and enantioselective reduction could provide an alternative potential for preparation of optically active anti-aldol-type compounds.

Revascularization using the short gastric vessels of the gastric tube after subtotal esophagectomy for intrathoracic esophageal carcinoma.

BACKGROUND: Maintaining sufficient blood flow to the substitutive organ after esophagectomy is essential to decrease the risk of anastomotic leakage. STUDY DESIGN: Forty-one patients underwent subtotal esophagectomy for intrathoracic esophageal carcinoma and reconstruction using the gastric tube. Additional vascular anastomosis between the short gastric vessels and the vessels in the neck was performed in 15 patients. Tissue blood flow was measured by laser Doppler flowmetry before and after vascular anastomosis. The incidence of anastomotic leakage in the revascularization group was compared with that in the remaining 26 patients. RESULTS: Venous anastomosis was performed in 14 patients and arterial anastomosis in 9. There was a significant increase in tissue blood flow after venous anastomosis alone (mean percent increase: 36%; p < 0.01), and after arterial and venous anastomoses (mean percent increase: 108%; p < 0.01). No anastomotic leakage was observed in the revascularization group; six patients (23.1%) in the control group had leakage (p < 0.05). Patients in the revascularization group started taking a meal 10.0 +/- 0.4 days postoperatively, compared with 15.1 +/- 1.8 days in the control group (p < 0.05). CONCLUSIONS: Additional vascular anastomosis in esophageal reconstruction after subtotal esophagectomy achieved good results. This procedure can reduce the risk of anastomotic leakage and may be useful for esophageal reconstruction.

Quantitative changes in amylase activity in the salivary glands, pancreas, saliva, and serum after administration of isoproterenol, pilocarpine, and acetylcholine.

Amylase activity in various tissues--i.e., submandibular/sublingual and parotid glands, the pancreas, saliva, and serum--in rats was measured after injection of isoproterenol, pilocarpine, and acetylcholine. All agents reduced amylase activity in the parotid gland and increased the enzyme activity in the submandibular/sublingual glands, in saliva and serum.

Activities of sulfatases for the degradation of acidic glycosaminoglycans in cultured skin fibroblasts from two siblings with multiple sulfatase deficiency.

Cultured skin fibroblasts from two siblings with multiple sulfatase deficiency (MSD) were assayed for the activities of sulfatases known to degrade acidic glycosaminoglycans (AGAG). There were iduronate sulfatase, arylsulfatase B, heparan sulfate (HS) sulfatase, N-acetylgalactosamine-6-sulfate sulfatase, HS-derived N-acetylglucosamine-6-sulfate sulfatase, and two keratan sulfate (KS)-derived N-acetylglucosamine-6-sulfate sulfatases. The activities of sulfatases required for the degradation of HS were reduced to a greater extent than those for the degradation of dermatan sulfate (DS), and those of sulfatases associated with basic defect of Morquio disease type A were moderately decreased or normal. On the other hand, urinary excretion of AGAG in both patients was increased about 10-fold compared to controls, and especially, the excretion of HS and DS was increased about 150-fold and 50-fold, respectively. Keratan sulfate was not detected. The results suggest that in patients with MSD the degradation of HS might be affected to a greater extent than that of DS.

Additional microvascular anastomosis in reconstruction after total esophagectomy for cervical esophageal carcinoma.

BACKGROUND: Maintaining sufficient blood flow to the substitute organ after total esophagectomy is essential for decreasing the risk of anastomotic leakage. Additional venous, or arterial and venous, anastomoses between the vessels of the gastric tube and the vessels in the neck after total esophagectomy are described for 11 patients with cervical esophageal carcinoma. METHODS: The tissue blood flow was measured by laser Doppler flowmetry before and after anastomosis. Venous anastomosis was performed for all 11 patients, and arterial anastomosis was added for 7 patients. RESULTS: A significant increase in tissue blood flow was observed after venous anastomosis alone (mean, 19%; P < 0.05) and after arterial and venous anastomoses (mean 43%; P < 0.01). There was no anastomotic leakage or hospital death. CONCLUSIONS: This procedure may reduce the risk of anastomotic leakage especially in the case of pharyngogastrostomy following total esophagectomy.

Effects of prolonged parotid duct ligation, parotidectomy and acute hepatitis of rats on amylase activity.

Amylase activity in the parotid gland was extremely reduced (to 2.4 per cent) by prolonged duct ligation but that in the pancreas and serum as similar to controls. The serum amylase activity of parotidectomized rats was remarkably reduced within a week of surgery, but recovered to approximately the previous level in 4 weeks. The amylase activities in the pancreas and liver were not different between control and parotidectomized rats. The serum amylase activity of rats with acute hepatitis (1 day after administration of D-galactosamine) decreased to 18.5 per cent compared with controls; however, the activity recovered to its original level after 4-7 days. There was no pancreatic amylase in the serum either of parotidectomized rats or those with acute hepatitis. These observations show that the parotid glands may not be the only source of serum amylase. Serum amylase is mainly derived from the parotid gland, but liver may contribute to serum amylase in compensation for the parotid glands.

Relationship between serum-amylase activity and intraductal pressures in the rat parotid and submandibular salivary glands after administration of pilocarpine or isoprenaline.

Ligation of the parotid ducts induced marked elevation of serum-amylase activity after administration of pilocarpine or isoprenaline. Secretion of saliva and serum-amylase activity increased most rapidly in the first 15 min after injection of pilocarpine, whereas isoprenaline induced a constant rate of secretion and serum-amylase activity over 120 min. Amylase activity in the serum was closely connected with intraductal pressure due to saliva secretion.

Transport to the bloodstream of amylase following retrograde infusion of amylase into the parotid glands in the rat.

Amylase secretion into parotid saliva was increased more by isoprenaline whereas the secretion rate of parotid saliva was increased more by pilocarpine. Retrograde infusion of parotid-amylase solution into excretory ducts of the parotid gland elevated the activity of the enzyme in the serum. Direct evidence that retrogradely-infused amylase reaches the blood from the parotid gland was obtained using amylase prepared from Bacillus subtilis. Pretreatment with pilocarpine accelerated the release of amylase into the blood. The elevation of intraductal pressure may be the primary mechanism for the release of amylase into the blood from the parotid glands.

Effects of prolonged duct ligation of the rat salivary glands on the activity of trypsin-like protease.

The effects of ligation on body weight, protease activity and hair kynurenine content were observed. The body weights of parotid gland(PT)-ligated and of combined submandibular (SM)-, sublingual (SL)- and PT-ligated rats were much less than those of sham-operated controls. However, the weights of SM,SL-ligated rats were similar to those of controls. The activity of trypsin-like protease in the duct-ligated submandibular gland was significantly reduced to 0.3-0.7 per cent of that in controls. Protein concentration also decreased in the duct-ligated submandibular gland. Kynurenine content in the hair of duct-ligated rats was higher than that in controls.

Effect of leupeptin, a protease inhibitor, on dopa decarboxylase and protease activities in homogenates of submandibular salivary glands from young rats.

Dopa-decarboxylase activity was high at 2-3 weeks of age, low at 5-6 weeks, and very low in adult rats; protease activity increased with age. Trypsin-like proteases, which decrease dopa-decarboxylase activity, were detected in the gland and were inhibited by a protease inhibitor, leupeptin. High concentrations (20 mg/ml) of leupeptin increased dopa-decarboxylase activity in the glands of adult rats to the level found at 5-6 weeks of age. The low activity of dopa decarboxylase in adult gland may be due in part to endogenous proteases.

Assay of trypsin-like protease in rat submandibular saliva and gland.

Trypsin-like protease activity was high in the submandibular gland, low in the pancreas, and absent in the liver, kidney and brain of adult rats. The alpha-sympathomimetic agent, norepinephrine, induced a marked secretion of the enzyme into submandibular saliva and the remaining protease activity within the gland decreased to about 64 per cent of control levels. Six peaks of protease activity, found by isoelectric focusing in intact submandibular glands, were also detected in the saliva of norepinephrine-stimulated rats. Isoprenaline also increased the protease activity in saliva, but far less so than norepinephrine. The parasympathomimetic agent, pilocarpine, had little effect on the secretion of the enzyme into saliva.

Mechanisms of increase in amylase activity in rat submandibular and sublingual glands after administration of pilocarpine.

The increase in parotid rather than pancreatic-type amylase activity in the submandibular and sublingual glands of rats caused by administration of pilocarpine was abolished or diminished when pilocarpine was injected into rats which had been parotidectomized, sympathectomized by superior cervical ganglionectomy or pretreated with reserpine. These results suggest that the increases in amylase activity in the submandibular and sublingual glands by pilocarpine are not due to increase in enzyme synthesis, but to uptake of enzyme released into the blood in large quantities from the parotid gland and that the release from the parotid gland by pilocarpine is primarily mediated by sympathetic nerves.

Retroperitoneal malignant fibrous histiocytoma surgically removed six times using a harmonic scalpel.

This paper describes a patient with retroperitoneal malignant fibrous histiocytoma who underwent surgical removal of the tumor 6 times. It is difficult to remove locally recurrent retroperitoneal malignant fibrous histiocytoma many times by repeat surgery, since the tumor tends to adhere tightly to the surrounding scar tissue. In the 5th and 6th operations we conducted on the patient, a harmonic scalpel was useful for separating the tumor from the scar tissue at the previous surgical site.

Technetium-99m-tetrofosmin imaging of lung cancer: relationship with histopathology.

Tc-99m-tetrofosmin is an agent to delineate cancer. To elucidate the usefulness of Tc-99m-tetrofosmin scintigraphy, we analyzed the relationship between the uptake of Tc-99m-tetrofosmin and histopathology in patients with lung cancer. SPECT studies were conducted twice: 15 minutes (early scan), and 60 minutes (delayed scan), after intravenous injection of 740 MBq Tc-99m-tetrofosmin. We calculated the retention index in order to evaluate the degree of Tc-99m-tetrofosmin retention in the primary tumor. The retention indices were significantly lower in squamous cell carcinoma than those of small cell carcinoma or adenocarcinoma. As the retention indices of Tc-99m-tetrofosmin were different in each histopathology, the index might play a part as a tumor marker of lung cancer.