Mutagenesis and reconstitution of middle-to-long-wave-sensitive visual pigments of New World monkeys for testing the tuning effect of residues at sites 229 and 233

The colour vision polymorphism of New World monkeys results from allelic variations of the middle-to-long-wave-sensitive (M/LWS) visual pigments. On the basis of sequence comparison, spectral differences among the alleles have been ascribed to amino acid residues at sites 180, 229, 233, 277, and 285. While the significant spectral effects have been demonstrated for sites 180, 277, and 285 by site-directed mutagenesis for a large number of vertebrate M/LWS pigments (the "three-site rule"), effects at sites 229 and 233 remain untested. Here we measured absorption spectra of the reconstituted M/LWS pigments from the tri-allelic squirrel monkey (Saimiri sciureus) and the mono-allelic owl monkey (Aotus trivirgatus). The peak absorption spectra (lambdamax) of Saimiri pigments were 532, 545, and 558 nm and that of Aotus pigment 539 nm, being consistent with the prediction from the three-site rule. Our site-directed mutagenesis for sites 229 and 233 showed that their mutational effects for lambdamax values were negligible. These results preclude the necessity of examining exon 4, encoding the residues at sites 229 and 233, of M/LWS pigment genes for colour-vision typing of New World monkeys.     

Involvement of 90K/Mac-2 binding protein in cancer metastases by increased cellular adhesiveness in lung cancer

90K/Mac-2 Binding Protein (M2BP) plays a role in regulation of immune responses and cell adhesive ability in patients with cancer and infectious diseases. We previously reported that M2BP was highly expressed in lung cancer and that immune responses to M2BP were increased in many patients with lung cancer. To determine the involvement of M2BP in metastatic processes of cancer progression, we examined the ability of M2BP DNA-transduced lung carcinoma cell lines to adhere to extracellular matrices. Although expressions of cell-surface integrins were not modulated in the M2BP transfectants, they showed increased adhesiveness to fibronectin and collagen IV. We next analyzed the serum levels of M2BP in patients with lung cancer and normal donors and the relationships between M2BP expression and clinicopathological factors in the patients. The M2BP level was markedly elevated in the patients and was strongly correlated with nodal involvement and clinical staging. To determine whether expression of M2BP by cancer cells is modulated in the environment of tumor-bearing hosts, M2BP expression in M2BP-positive QG56 cells following exposure of the cells to pro-inflammatory cytokines was examined. The M2BP expression in QG56 cells was up-regulated by many of the cytokines that activate host protective immunity. The findings in this study suggest that M2BP plays a role in cancer metastasis by increased adhesiveness of cancer cells and that M2BP is increasingly produced even in a state of exposure to the host immune system. This molecule may be useful as a predictive factor of disease progression in lung cancer.     

Medical therapy corresponding to the morbid state of hyperlipidemia

An appearance of statin did not only cause fall of onset rate and the death rate of coronary arteries disease, but also a fall of the total death rate. These results are restraint of cardiovascular event, development and regression restraint of plaque, function improvement of plaque constitution cells, resulting in the decrease of the total death. However, most of the clinical lipid intervention trial were European and USA results, and there was a difference of a quantity of dosage, too, and results in a Japanese were waited for. Results of a clinical lipid intervention trials in a Japanese appeared by degrees, too though a little too late. From those results, it is proposed that hyperlipidemia is managed whether there are several risk factors of atherosclerosis except hyperlipidemia. Needless to say, it is for the greatest purpose of hyperlipidemia treatment to prevent the development and progression of atherosclerosis. Therefore, it is necessary to treat hyperlipidemia after having fully understood the function mechanism of lipid-lowering agents and after having thought about the morbid state of a patient. In addition, on the occasion of a combination treatment it is necessary to administer it after having understood the each other's function mechanism.     

The development of new method for the assessment of perfusion reserve using split dose iodine-123-IMP SPECT: one-day protocol by modified ARG method

Evaluation of the regional cerebrovascular reactivity (rCVR) to a cerebral vasodilatory stimulus is important in the investigation of patients with ischemic cerebrovascular disease. We devised a simplified one-day protocol technique using [123I]N-isopropyl-p-iodoamphetamine (IMP) autoradiography (ARG) with SPECT. To validate the accuracy of IMP-ARG for quantifying rCVR to acetazolamide, we compared rCVR measured using IMP-ARG with rCVR calculated using IMP split dose method of microsphere model. Twenty patients with chronic steno-occlusive disease in a unilateral major cerebral artery underwent 123I-SPECT. On rCBF SPECT image above 3.5 cm from OM line, large cortical regions of interest (ROI) was bilaterally determined for bilateral middle cerebral artery and anterior cerebral artery. Based on rCBF values in each ROI, rCVR to acetazolamide was calculated. Significant correlation was observed between rCVR values obtained using IMP-ARG and microsphere model IMP methods in the 80 ROIs examined in the 20 patients (r = 0.72; p < 0.001). The result demonstrated that [123I]IMP-ARG split dose method can quantify rCVR non-invasively in a short time.     

Did the widespread use of long-acting calcium antagonists decrease the occurrence of variant angina?

BACKGROUND: We have not often encountered variant angina (VA) since the use of long-acting calcium antagonists (L-CAs) became widespread. OBJECTIVES: This study examined the frequency of VA retrospectively. METHODS: and results: We diagnosed angiographically confirmed coronary spastic angina (CSA) in 349 consecutive patients using selective spasm provocation tests from January 1991 to December 2002. During this period, 3,148 diagnostic cardiac catheterizations and 1,515 selective spasm provocation tests were performed. Seventy-four of these 349 patients (21.2%) had VA. Coronary spasms were defined as transient luminal narrowings of > 99%, and VA was defined as an ST elevation during spontaneous attacks or noninvasive stress tests. We classified the 12 years of the study into four periods of 3 years each. No tendency to decrease for the ratio of the number of patients with CSA and the number of selective spasm provocation tests was observed among the four time periods (18%, 24%, 32%, and 23%, respectively). However, the number of patients with VA (28, 33, 9, and 4) and the VA/CSA ratio (32%, 28%, 14%, and 5%, respectively) in the four group significantly decreased. The frequency of administration of calcium antagonists (CAs) before hospital admission (49% vs 33%, respectively; p < 0.05) was significantly higher in the last time period (from 2000 to 2002) than in the first period (from 1991 to 1993). L-CAs were administered in > 90% of CSA patients who had been medicated with CAs before hospital admission in the last period (from 2000 to 2002), while L-CAs were administered in only 20% in the former period (from 1991 to 1993). The administration of statins and angiotensin-converting enzyme inhibitors/angiotensin receptor blockers before hospital admission gradually increased according to the period passed, but not significantly. CONCLUSION: The frequency of VA has decreased in Japan, possibly due to the widespread use of therapy with L-CAs.     

Postoperative Evaluation of Pylorus-Preserving Procedures Compared with Conventional Distal Gastrectomy for Early Gastric Cancer

We evaluated postoperative function in 98 patients who underwent surgery for early gastric cancer between 1995 and 1998 to compare the results of pylorus-preserving procedures to those of conventional distal gastrectomy with Billroth I (B-I). The pylorus-preserving procedures included endoscopic mucosal resection (EMR), performed in 12 patients; local resection (Local), performed in 14 patients; segmental resection (Seg), performed in 8 patients; and pylorus-preserving gastrectomy (PPG), performed in 19 patients. B-I was performed in 45 patients. The nutritional status and serum albumin (Alb) levels after PPG, the hemoglobin (Hb) levels after EMR, Local, and PPG, and the present/preoperative body weight ratios after EMR, Local, Seg, and PPG were superior to those after B-I. The time before oral intake was recommenced after EMR and Local, the volume of oral intake tolerated after EMR, Local, Seg, and PPG, and the postoperative hospital stay after EMR were all superior to those after B-I. Moreover, significantly fewer patients suffered reflux symptoms after EMR, Local, and PPG, abdominal fullness after EMR, and early dumping syndrome after EMR, Local, and PPG than after B-I. There was also less evidence of gastritis after EMR, Local, and PPG, and of bile reflux after EMR, Local, and PPG, than after B-I. These findings indicate that pylorus-preserving procedures may result in a better postoperative quality of life for selected patients with early gastric cancer. Received: September 28, 2000 / Accepted: March 6, 2001     

Orally administered L-ornithine elevates brain L-ornithine levels and has an anxiolytic-like effect in mice

Intracerebroventricular injection of L-ornithine has demonstrated sedative and hypnotic effects in neonatal chicks exposed to acute stressful conditions. However, whether orally administered L-ornithine can reduce acute mental stress remains to be defined. To clarify the nutritional importance of L-ornithine in controlling the stress response, in Experiment 1 we first investigated whether orally administered L-ornithine can be transported into the brain of mice. Mice were orally administered L-ornithine (3 mmol/water 10 ml/kg, per os). L-Ornithine levels were significantly elevated in the cerebral cortex and hippocampus at 30 and 60 minutes post-administration. In Experiment 2, the effect of orally administered L-ornithine (0, 0.1875, 0.75 and 3 mmol/water 10 ml/kg, per os) on anxiety-like behavior in mice exposed to the elevated plus-maze test was examined at 30 minutes post-administration. There was a significant increase in the percentage of time spent and entries in the open arms in the group receiving 0.75 mmol of L-ornithine compared to the control group. Furthermore, locomotion activity in a novel environment was not significantly changed between the control group and 0.75 mmol of L-ornithine group in Experiment 3. Therefore, it appears that orally administrated L-ornithine is bioavailable to the rodent brain and reduces anxiety-like behavior as demonstrated by the elevated plus-maze test.     

Oral administration of chicken breast extract increases brain carnosine and anserine concentrations in rats

Carnosine (beta-alanyl-L-histidine) and its derivative anserine (beta-alanyl-1-methyl-L-histidine) are antioxidants and putative neurotransmitters in the brain. These dipeptides are rich in the commercially available supplement chicken breast extract (CBEX). To clarify the effects of CBEX on the brain, we examined whether single oral administration of CBEX (20 ml/kg) affects brain dipeptide and free amino acid concentrations in male Wistar rats. CBEX significantly and time-dependently increased carnosine and anserine levels in the plasma (at 120 min after injection, increase rates were 2976 and 4142%, respectively), hippocampus (64 and 78%), and hypothalamus (188 and 120%), but not in cerebral cortex. Significant and time-dependent increases in citrulline in the hippocampus (49%) and hypothalamus (41%) demonstrated generation of nitric oxide due to the increased carnosine and/or anserine levels in these brain regions. These findings suggest that CBEX modifies brain functions by increasing levels of these dipeptides.