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Parkinson's disease is neuropathologically characterized by the presence of Lewy bodies, whose major component is alpha-synuclein. We had previously generated transgenic mice that expressed human alpha-synuclein carrying an Ala53Thr point mutation (halpha-syn140m) under the control of the rat tyrosine hydroxylase promoter and found that halpha-syn140m was localized not only in the cytoplasm but also in the nuclei of mesencephalic dopaminergic neurons. In the present study, we carried out immunohistochemical analysis of the brain of Tg mice using anti-PSer129, an antibody that specifically recognizes alpha-synuclein phosphorylated at Ser129. The antibody detected only phosphorylated halpha-syn140m, whereas phosphorylation of endogenous alpha-synuclein, if any, was below the detection limit of the method employed. The analysis showed that approximately one-third of the halpha-syn140m-positive neurons in the midbrain of heterozygous Tg mice were concomitantly reactive to anti-PSer129. The ratio almost doubled in homozygotes, indicating that the phosphorylation level depends directly on the amount of substrate. In addition, the ratio did not change at least up to 48 weeks of age. These data strongly suggest that halpha-syn140m underwent constitutive phosphorylation and that the phosphorylation level was maintained to a certain level until the aged stages. Remarkably, halpha-syn140m localized in the nuclei seemed to be preferentially phosphorylated compared with that in the cytoplasm. Among kinases that have been reported to be involved in the phosphorylation of alpha-synuclein, the beta subunit of casein kinase-2 was detected in the nuclei by immunohistochemistry. These data imply that at least casein kinase-2 is involved in the phosphorylation of halpha-syn140m in the Tg mice., (Copyright (c) 2007 Wiley-Liss, Inc.)

The purpose of this study was to determine whether primary sensory neurons that innervate the uterus receive convergent input from the colon. To test this, in the rat, cell bodies of colonic and uterine dorsal root ganglia were retrogradely labeled with fluorescent tracer dyes microinjected into the colon/rectum and bilaterally into the uterine horns. Ganglia were harvested, cryoprotected and cut into 20 microm slices to identify positively stained cells for fluorescent microscopy. Up to 5% of neurons were colon-specific or uterus-specific, and 10-15% of labeled ganglion neurons innervate both viscera in the L1, L2, L6 and S1-S3 levels. These results suggest a novel form of visceral sensory integration in the dorsal root ganglion that may underlie comorbidity of many functional pain syndromes.

Two methoxy-substituted TQEN (N,N,N',N'-tetrakis(2-quinolylmethyl)ethylenediamine) derivatives, T(MQ)EN (N,N,N',N'-tetrakis(6-methoxy-2-quinolylmethyl)ethylenediamine) and T(TMQ)EN (N,N,N',N'-tetrakis(5,6,7-trimethoxy-2-quinolylmethyl)ethylenediamine), have been prepared, and their fluorescence properties with respect to Zn2+ coordination were investigated. Introduction of a methoxy substituent at 6-position of the quinoline ring enhances the fluorescence intensity by 10-fold, and the three methoxy substituents in the 5,6,7-positions afford significant enhancement of the long-wavelength component of the fluorescence of zinc complex. The substituents did not alter the binding affinity of these compounds toward zinc ion significantly. T(MQ)EN was proved to be effective in detection of zinc ion in cells by fluorescent microscopy.

We examined the development of cardiac hypertrophy in juvenile visceral steatosis (JVS) mice, a model of systemic carnitine deficiency, by varying the amount of lipid in the diet. Cardiac hypertrophy was markedly attenuated by decreasing soy bean oil (SBO) from 5% (w/w) to 1%. Triglyceride contents of the ventricles of JVS mice fed 1% SBO were significantly lower than in JVS mice fed 5% SBO. The addition of medium-chain triglycerides metabolically utilized by JVS mice did not affect the development of cardiac hypertrophy. On the other hand, the mRNA levels of atrial natriuretic peptide and skeletal alpha-actin, which are related to cardiac hypertrophy, were also attenuated by decreasing lipid in the diet. Adenylate energy charge and creatine phosphate in the heart of JVS mice at the early stage of hypertrophy were not significantly different from control mice given the same laboratory chow (4.6% of lipid). Although urinary prostaglandin F(2alpha) levels were found to be increased in JVS mice at 15 days of age when they developed cardiac hypertrophy, administration of aspirin was not efficacious. We, therefore, propose that the proportion of lipid in the diet is important in the development of cardiac hypertrophy in carnitine-deficient JVS mice, and that this is not related to prostaglandin formation.

The effects of systemic administration of bovine beta-casomorphin-5 (Tyr-Pro-Phe-Pro-Gly), a mu-opioid receptor agonist derived from milk beta-casein, on spontaneous alternation behavior in the Y-maze (spatial short-term memory) and step-down-type passive avoidance response (non-spatial long-term memory) were investigated in mice. Intraperitoneal (i.p.) administration of beta-casomorphin-5 (0.1-20 mg/kg) did not have a significant effect on either spontaneous alternation behavior or passive avoidance response. However, a low dose (1 mg/kg, i.p.) of beta-casomorphin-5 improved scopolamine (1 mg/kg, s.c.)-induced impairment of spontaneous alternation behavior and passive avoidance response. Pretreatment with intracerebroventricular injections of beta-funaltrexamine (a mu-opioid receptor antagonist, 0.1 microg/mouse) and naloxonazine (a mu(1)-opioid antagonist, 5 microg/mouse), which did not improve scopolamine-induced impairment, prevented the ameliorating effect of beta-casomorphin-5 on scopolamine-induced impairment of passive avoidance response. These results indicated that systemic administration of a low dose (1 mg/kg, i.p.) of beta-casomorphin-5 improves the disturbance of learning and memory resulting from cholinergic dysfunction through central mediation involving mu(1)-opioid receptors.

A new heptadentate N6-O1 ligand, N,N,N',N'-tetrakis(2-quinolylmethyl)-2-hydroxy-1,3-propanediamine (Htqhpn), was synthesized and used to generate compounds with linearly ordered MnIIMnIIIMnIIIMnII tetranuclear cores. This is the lowest valent tetranuclear manganese complex that exhibits a (mu2-O)2Mn2 core in the molecule. The electron paramagnetic resonance and magnetic measurements of these tetranuclear complexes suggest moderately strong antiferromagnetic coupling for the central MnIII2 core, with weak coupling between the MnII and MnIII centers.

QT dispersion is defined as the difference in QT interval among the different leads of the standard 12-lead electrocardiogram and reflects inhomogeneity of myocardial repolarization. Dispersion of repolarization is an important electrophysiologic feature that is considered fundamental for the initiation of ventricular fibrillation. However, no data exist regarding the diurnal variation of QT dispersion measured from simultaneous 12-lead recording in chronic heart failure patients. The aim of this study was to identify diurnal variation in QT dispersion in patients with chronic heart failure. QT dispersion was measured in the 12-lead standard electrocardiogram in 11 patients with chronic heart failure. QT dispersion in these patients was increased in the afternoon compared to the morning. It is concluded that QT dispersion has a clear diurnal variation in patients with chronic heart failure. These findings have potentially significant implications for therapy and prevention of sudden cardiac death in patients with chronic heart failure.

A new fluorescent sensor for zinc that binds 1 equivalent of zinc ion, N,N,N',N'-tetrakis(2-quinolylmethyl)ethylenediamine (TQEN), has been prepared and characterized. Zinc-bound TQEN exhibits fluorescence around 383 nm upon excitation at 317 nm, while free TQEN emits very weak fluorescence. UV-Vis and 1H NMR spectral changes also detected the binding of TQEN with zinc ion. The crystal structure of zinc complex with TQEN was determined by X-ray crystallography and compared with that of the TPEN-Zn complex (TPEN =N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine). The binding affinity of TQEN with zinc ion is very high (Kd < 1 microM in aqueous dmf solution). Competition experiments reveal that the zinc-binding affinity of TQEN is lower than the parent, strong metal ion chelator TPEN, and comparable to EGTA (EGTA = ethylene glycol-bis(2-aminomethyl)-N,N,N',N'-tetraacetic acid).

Anthocyanins are the chemical components that give the intense color to many fruits and vegetables, such as blueberries, red cabbages and purple sweet potatoes. Extensive studies have indicated that anthocyanins have strong antioxidant activities. To investigate the mechanism of anthocyanidins as an anticancer food source, six kinds of anthocyanidins representing the aglycons of most anthocyanins, were used to examine their effects on tumor promotion in mouse JB6 cells, a validated model for screening cancer chemopreventive agents and elucidating the molecular mechanisms. Of the six anthocyanins tested, only those with an ortho-dihydroxyphenyl structure on the B-ring suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cell transformation and activator protein-1 transactivation, suggesting that the ortho-dihydroxyphenyl may contribute to the inhibitory action. Delphinidin, but not peonidin, blocked the phosphorylation of protein kinases in the extracellular signal-regulated protein kinase (ERK) pathway at early times and the c-Jun N-terminal kinase (JNK) signaling pathway at later times. p38 kinase was not inhibited by delphinidin. Furthermore, two mitogen-activated protein kinase (MAPK) specific inhibitors (SP600125 for JNK and UO126 for ERK) could specifically block the activation of JNK and ERK and cell transformation. Those results demonstrate that anthocyanidins contribute to the inhibition of tumorigenesis by blocking activation of the MAPK pathway. These findings provide the first molecular basis for the anticarcinogenic action of anthocyanidins.

The effects of intracerebroventricular (i.c.v.) injection of bovine beta-casomorphin-5 (beta-CM-5: Tyr-Pro-Phe-Pro-Gly), a micro-opioid agonist derived from milk beta-casein, on step-down type passive avoidance tasks were investigated in mice. Intracerebroventricular administration of a high dose (10 microg) of beta-CM-5 produced a significant decrease in step-down latency. beta-Funaltrexamine (5 microg, i.c.v.) almost completely reversed the beta-CM-5-induced shortening of step-down latency, although neither naltrindole (5 ng, i.c.v.) nor nor-binaltorphimine (5 microg, i.c.v.) had any significant influence on the effect of beta-CM-5. Meanwhile, a low dose (0.5 microg, i.c.v.) of beta-CM-5 inhibited scopolamine (1 mg/kg)-induced impairment of passive avoidance response. These results indicated that a high dose of beta-CM-5 induces amnesia, whereas a low dose ameliorates scopolamine-induced amnesia.

In cyanobacterium Synechococcus sp. PCC 7942, SmtB, functioning as the sensor to heavy-metal ions (notably Zn-ion) in the dimer form, represses the transcription of smtA gene encoding metallothionein-like protein. There are two recognition DNA sequences in the operator/promoter region of smtA gene, and the binding of SmtB with Zn-ions reduces the affinities to these sequences. This induces the activation of smtA transcription. In this study, we have studied the functional differences between these two recognition DNA sequences, with spectroscopic (hetero-nuclear NMR and UV-resonance Raman) and biochemical methods. On the basis of the results obtained here, we clarified the regulation mechanism of the smtA expression. Similar regulatory system for the tolerance to Zn-ion stress is found in another cyanobacterium. We also found that the functionally important differences between these two systems are mainly due to the structural differences of recognition DNA sequences.

SmtB is a trans-acting dimeric repressor of the metal-regulated smtA gene, and the release of SmtB from the smtA operator/promoter region is essential for the tolerance to Zn(2+) stress by SmtA expression. Gel retardation assaying demonstrated that different sizes of SmtB-DNA complexes were formed depending on the DNA sequences, and the amounts of these complexes decreased in the presence of Zn(2+). Here, we present the first direct evidence that Zn(2+ )(>4 micro M) inhibits the SmtB-DNA complex formation in vitro, which ensures the physiological functions of SmtB as a Zn(2+) sensor and a transcription factor.

Tolerance to the heavy-metal ion stress in cyanobacterial cell is regulated by the transcriptional repressor SmtB that senses the Zn/Cd concentration in the cell. SmtB regulated the transcriptional level of class II metallothionein, SmtA. There are two recognition DNA sequences (Bbs1 and Bbs2) in the operator/promoter region of smtA gene. To clear the functional meaning of the presence of these two sequences, we have compared the affinities of native and point-mutated SmtBs to these two sites. We also have compared the sizes of the protein-DNA complexes being formed with these two sites. SmtB forms protein-DNA complex in an unique size with Bbs1, in three different sizes with Bbs2. We have further designed the modified Bbs1 and Bbs2 sequences, and the results obtained with these sequences indicate that the differences observed between the cases of Bbs1 and Bbs2 are caused by the presence of direct repeat sequence and the differences in the linker sequences.

We have cloned the gene for chicken 2',5'-oligoadenylate synthetase (ChOAS) by the method of polymerase chain reaction with use of ChOAS cDNA sequence. The ChOAS gene is composed of five introns and six exons containing all of the sequence of the ChOAS cDNA from the start to the stop codon. The first five exons of ChOAS gene which encode the OAS catalytic domain have a similar structure to HuOAS1 gene including the exon-intron boundaries. However, the length of introns of ChOAS gene is only 1/7 of those of HuOAS1 gene. The sixth exon of the ChOAS gene encodes the ubiquitin-like (UbL) domain of two consecutive sequence (UbL1 and UbL2) homologous to ubiquitin. ChOAS encoded in a single copy gene has at least two alleles, OAS(*)A and OAS(*)B. The differences between these two alleles are in the sixth exon of the gene; a 96-nucleotide sequence in the UbL1 portion of OAS(*)A is deleted from OAS(*)B. No OAS(*)B gene was detected in nine lines of chickens tested other than Leghorns. Almost the same levels of ChOAS-A and -B proteins induced physiologically in erythrocytes were detected in infant chickens (2-week-old), but in grown-up chickens (6-month-old) the level of erythrocyte OAS-B was markedly reduced in most of B/B chickens. Thus, the UbL domain of ChOAS is responsible for the maintenance of the OAS level in the tissue.

Improvement of the methods for oligonucleotide delivery into cells is necessary for the development of antisense therapy. In the present work, a new strategy for oligonucleotide delivery into cells was tested using cationic peptides as a vector. At first, to understand what structure of the peptide is required for binding with an oligonucleotide, several kinds of alpha-helical and non-alpha-helical peptides containing cationic amino acids were employed. As a result, the amphiphilic alpha-helix peptides were best for binding with the oligonucleotide, and the long chain length and large hydrophobic region in the amphiphilic structure of the peptide were necessary for the binding and forming of aggregates with the oligonucleotide. In the case of non-alpha-helical peptides, no significant binding ability was observed even if their chain lengths and number of cationic amino acid residues were equal to those of the alpha-helical peptides. The remarkable ability of oligonucleotide delivery into COS-7 cells was observed in the alpha-helical peptides with a long chain length and large hydrophobic region in the amphiphilic structure, but was not observed in the non-alpha-helical peptides. It is considered that such alpha-helical peptides could form optimum aggregates with the ODN for uptake into cells. Based on these results, the alpha-helical peptide with a long chain length and large hydrophobic region is applicable as a vector for the delivery of oligonucleotides into cells.

The autopsy of a 76-year-old Japanese female patient, which revealed thymic carcinoma with various tumor markers such as NSE, CYFRA, and CA-125, is presented. The patient died from hepatic failure because the liver was overtaken by the tumors. At autopsy, the thymic carcinoma was found to have metastased only in the liver. From microscopical analyses and electron microscopical findings, we diagnosed poorly differenciated squamous cell carcinoma of thymic origin. In the histochemical analyses, the tumor cells were positively stained in CA 125, CA 19-9, EMA, NSE, AE 1, AE 3, CEA, S-100, glimerius and Bcl-2. These date suggest that the tumor cells produced various tumor markers. In 222 autopsy cases of thymic malignant tumor observed in Japan over a period of 4 years, the dominant pathohistological image was squamous cell carcinoma. It is interesting that the greatest number of combined malignant tumors with thymic malignancies were thyroid papillary carcinomas.

Two xylanase genes (xyn1 and xyn2) were amplified by the polymerase chain reaction (PCR) technique from first-strand cDNA prepared from mRNA of Trichoderma reesei QM9414. The genes were located under the human cytomegalovirus gene promoter (CMVp) on copy-number-controlled plasmids (pTLxyn1 and pTLxyn2). When both plasmids were introduced into Schizosaccharomyces pombe, functional xylanases (XYN I and XYN II) were secreted by the recombinant yeasts. The secreted XYN I protein had a molecular mass of 21 kDa whereas XYN II was produced as two molecular forms with sizes of 21 and 28 kDa, the former being not glycosylated and the latter N-glycosylated. XYN I was secreted in the culture medium at a level of about 25 microg/ml and XYN II at about 170 microg/ml. The recombinant xylanases had the same characteristics with respect to the effects of temperature and pH on the enzyme activity as the native ones.

Blood glucose and ketone body (3-hydroxy-butyrate) concentrations were measured in 8 children (4.2 +/- 2.8 years old) during the repair of lipomyelomeningocele (9.0 +/- 2.6 hours) to elucidate whether lipid mobilization can be prevented by glucose infusion at the rate of 0.2 +/- 0.05 g.kg-1.h-1. The operation was followed in about 4 hours by a significant increase in blood glucose and ketone body concentrations, with their maximum value of 195 +/- 75 mg.dl-1 and 656 +/- 75 mumol.l-1, respectively (P < 0.05), as compared with the preoperative level. The lipid mobilization soon returned to normal state by increasing the rate of glucose infusion, as indicated by ketone body level of 92 +/- 58 mumol.l-1 at the end of operation. These results show that glucose infusion rate should be adjusted based on serial analysis of both blood sugar and ketone body concentrations in the anesthetic management of children undergoing a long operation.

A case of mixed connective tissue disease (MCTD) in a female with positive ribonucleoprotein antibody and overlapped manifestations of systemic lupus erythematosus and dermatomyositis is presented. During the last 9 years of her 13 year clinical course, she repeatedly manifested abdominal distension, pneumoperitoneum and malabsorption syndrome. She died, aged 54 years, of respiratory failure secondary to pulmonary compression from marked abdominal distention. Autopsy revealed esophageal fibrosis with ulceration and the typical appearance of pneumatosis cystoides intestinalis (PCI), in association with atrophy and fibrosis of the inner circular muscle layer of the small bowel. The association of PCI with MCTD is reviewed briefly.

We studied the effects of prostaglandin E1 (PGE1) on hand-foot temperature gradients (foot temperature-hand temperature) induced by lumbar epidural anesthesia. Thirty-six ASA class 1 patients undergoing rotatory acetabular osteotomy were randomly assigned to receive 0 gamma (n = 13), 0.005 gamma (n = 10) and 0.02 gamma (n = 13) of PGE1. Thermometric probes were placed on the palm and sole of each patient. Bladder temperature was also monitored to evaluate the change of core temperature. Epidural catheter was indwelled at the level between L2 and L3 prior to induction and 2 % lidocaine was administered epidurally. All patients were warmed by water blanket at 37 degrees C and fluids for infusion warmed at 37 degrees C. Humidity and moisture exchanger was placed in breathing circuit. Hand-foot temperature gradients of patients given 0 and 0.005 gamma of PGE1 increased to 2.8 +/- 1.5 and 2.3 +/- 2.3 degrees C at extubation, respectively. In contrast, temperature gradient of patients given 0.02 gamma of PGE1 was 0.0 +/- 1.8 degrees C (P < 0.01). There was no difference among the values of bladder temperature in each group. The effects of PGE1 on mean arterial pressure, heart rate and central venous pressure were minimal. These data suggest that 0.02 gamma of PGE1 administration is effective to reduce hand-foot temperature gradient during epidural anesthesia.

Aldolase deficiency of red blood cell is a rare cause of hereditary hemolytic anemia and now there exists only three patients in the world. We had a 24-year-old man operated on for gallbladder stone secondary to this uncommon disease. He underwent a cholecystectomy under general anesthesia combined with thoracic epidural block, using isoflurane, fentanyl, vecuronium, midazolam and lidocaine. During the surgery serum concentrations of bilirubin, free hemoglobin and LDH showed no change, suggesting a lower incidence of drug-induced hemolysis in the case of aldolase deficiency than in other enzyme deficiency. This fact also provides a useful guide to the choice of anesthetics and related agents. In the postoperative period, however, we found a hemolytic response to fever with a drop in hemoglobin level to 2.5 g.dl-1. Aldolase activity of his red cell is heat labile and an increase in body temperature may aggravate a disturbance in the glycolytic pathway leading to hemolytic crisis. It is thus important to prevent the body temperature from rising when a patient is suffering from hemolytic anemia due to red cell aldolase deficiency.

We have measured blood concentrations of glucose, 3-hydroxybutyrate (3-OHBA), non-esterified fatty acid (NEFA) and lactate before and after glucose administration in 105 patients to determine the optimal glucose dose during the period of preoperative fasting state. The patients, scheduled for laparotomy in the afternoon, received fluid infusion containing an arbitrary glucose dose for 4 hours prior to surgery. The glucose dose showed a negative correlation with the ratio of 3-OHBA or NEFA prior to and after glucose administration (r = 0.40, r = 0.41 respectively, P < 0.05). There was no correlation between lactate ratio and glucose dose (r = 0.06). Glucose administration more than 0.4 g.kg-1 significantly suppressed 3-OHBA production (P < 0.05). Hyperglycemia (> 200 mg.dl-1) developed more often in the groups given 0.5 g.kg-1 or more glucose (P < 0.05), while significant hypoglycemia (< 60 mg.dl-1) occurred in the groups given less than 0.2 g.kg-1 of glucose (P < 0.05). This study demonstrates that preoperative infusion of glucose 0.4-0.5 g.kg-1 is useful to maintain the energy metabolism during the fasting state in patients scheduled for operation in the afternoon.

We studied the effects of prostaglandin E1 (PGE1) on renal function during and after deliberate hypotension induced by epidural anesthesia. Twenty-seven ASA class 1 patients undergoing rotatory acetabular osteotomy were divided into three groups according to the dose of PGE1. Deliberate hypotension was induced and maintained during the surgery by epidural anesthesia combined with general anesthesia. PGE1 increased urine output and creatinine clearance (Ccr) during hypotension in dose dependent manner, and the changes were statistically significant compared with control at the dose of 0.02 micrograms.kg-1 x min-1 (gamma). PGE1 also tended to increase postoperative urine output and Ccr. Regardless of PGE1 administration, urine NAG/Cr ratio tended to increase during and after deliberate hypotension. Though total dose of local anesthetics was attenuated by PGE1, serum catecholamine levels of all patients were within normal ranges. In conclusion, PGE1 0.02 gamma is effective to maintain urine output during deliberate hypotension with epidural anesthesia. However, PGE1 is unlikely to prevent renal tubular damage at the dose less than 0.02 gamma.

Ninety total hip replacements (THRs) performed by one surgeon were reviewed to compare the effects of different anesthetic techniques on the perioperative bleeding. Half of the THRs were performed under either combined spinal and epidural (CSE) anesthesia with lidocaine or general anesthesia with N2O/O2, vecuronium, enflurane or isoflurane. Deliberate hypotensive technique was not employed in any THR. All patients were female and had suffered from osteoarthritis of the hip joints. The patients in the two anesthesia groups were similar as to age, body weight, height, perioperative hemoglobin level, duration of anesthesia and operation, and blood transfusion requirement. A positive linear correlation existed between total operative blood loss and operation time in each group (P < 0.01). The rate of operative blood loss was significantly higher in the CSE anesthesia (6.2 +/- 3.0 ml.min-1) than in the general anesthesia (5.1 +/- 2.0 ml.min-1), while the amount of blood loss itself was without statistical difference. A significant increase in total volume of perioperative blood loss was also observed in the group with CSE anesthesia (1520 +/- 90 ml) compared with the general anesthesia group (1279 +/- 58 ml). These results show that spinal and/or epidural anesthesia can not always lead to reductions in operative blood loss during THR.

We have studied common problems associated with the anesthetic management of Forestier's disease in 8 patients undergoing neurosurgery. This disease is a systemic degenerative disorder and is characterized by osteo-spur formation at anterior spinal body. In two of eight patients the trachea was intubated with great difficulty even by well-trained anesthesiologist. Its cause could be found not only in limited cervical mobility but also in constrained epiglottic elevation, suggesting that Forestier's disease requires special anesthetic considerations. It is thus important to predict the possibility of difficult tracheal intubation on both physical examination and the preoperative X-ray photography.

We have studied the concentrations of catecholamines in collected autologous blood. The measured levels of the blood samples from Cell-stat collecting chamber were epinephrine 4.74 ng.ml-1 and norepinephrine 2.39 ng.ml-1. First wash of collected blood with 700 ml of saline diluted the catecholamine concentrations to epinephrine 3.13 ng.ml-1 and norepinephrine 1.6 ng.ml-1. The concentrations of catecholamines after second wash were epinephrine 2.19 ng.ml-1 and norepinephrine 1.11 ng.ml-1. These values were three to twenty folds of normal ranges even after the second wash, and still the same levels as intraoperative plasma catecholamine (the measured values; epinephrine 0.81-2.81 ng.ml-1 and norepinephrine 0.96-3.15 ng.ml-1). Since platelets actively concentrate catecholamines during their life span, the destruction of platelets by suction or centrifugation may probably play the most important role in the elevation of catecholamine concentrations in the collected autologous blood. We concluded that intraoperative autotransfusion in the resection of pheochromocytoma is likely to result in the elevation of systemic blood pressure by catecholamines of the transfused blood.

The pros and cons as to which anesthesia is more beneficial, either spinal or epidural, prompted us to perform combined spinal and epidural block in the elderly undergoing lower limb surgery. The selected epidural space was entered with a 17-gauge Tuohy needle and a longer 26-gauge spinal needle was passed through it and into the subarachnoid space. Following the injection of required dose of 2% preservative-free lidocaine (isobaric), spinal needle was withdrawn and an epidural catheter was inserted. We could use this combined technique on 17 patients older than 80 yr (mean age; 84.5 +/- 3.9 years) with satisfactory results and without any serious cardiovascular change, as with 17 middle-aged patients (57.5 +/- 5.2 years). This combination of techniques provides a rapid onset and reliability of spinal block with high quality analgesia by supplementation through the epidural catheter during and after surgery. In the orthopaedic procedures on the lower limbs, combined spinal and epidural block is more useful even for the elderly over the age of 80 yr than spinal or epidural anesthesia alone.

In a double blind test, the effects of ONO 3708, thromboxane A2 (TXA2) receptor antagonist, on hemodynamic responses after neutralization of heparin by protamine, were evaluated in 19 patients undergoing coronary artery bypass graft. Severe circulatory disturbances were not observed in all patients after intravenous administration of protamine (3 mg.kg-1) over 5 minutes, and in particular ONO 3708 (2.5 micrograms.kg-1.min-1 given with continuous infusion) group (n = 10) showed no deleterious hemodynamic responses to protamine. On the other hand, in placebo group (n = 9) the mean pulmonary artery pressure (mPAP) and the mean pulmonary/systemic artery pressure ratio (Pp/Ps) increased significantly, immediately following protamine administration, compared with the baseline values and ONO 3708 group. The results suggest that pulmonary hypertension after protamine is associated with TXA2 release and that ONO 3708 is useful to avoid this reaction.

With the development of medical ultrasonics, anesthesiologists have recognized the usefulness of transesophageal echocardiography as an intraoperative monitoring. Intraoperative transesophageal echocardiography is now used to monitor left ventricular function and to confirm the result of surgical correction immediately after cardiac operation. In this study, we evaluated the detectability of air embolism and the effect of hemodynamic changes on physiological mitral regurgitant flow by transesophageal Doppler echocardiography (TEDE). TEDE was more sensitive than precordial Doppler ultrasound in detecting intracardiac air. TEDE could detect air inflow to the right atrium earlier than alterations in pulmonary hemodynamics. The monitoring of intracardiac air by TEDE minimized the complication by air embolism. Regarding physiological mitral regurgitation, the degree of regurgitation increased with the elevation of BP in 70% of patients. An increase in afterload seemed to cause accidental volume loading on the left heart resulting in physiological mitral regurgitation. Intraoperative monitoring of regurgitant flow by TEDE was useful for the anesthesiologist as a guide of hemodynamic management. Transesophageal echocardiography is a useful intraoperative monitoring method regardless of its several disadvantages. With further improvement of mechanical device, transesophageal echocardiography will be more often and safely performed in operating room.

Metabolic parameters and anaphylatoxin activities in mixed venous blood were measured in 16 patients undergoing abdominal aortic reconstructive surgery to study the mechanism of pulmonary hypertensive response after aortic declamping. This reaction was confirmed by a rise in ratio between mean pulmonary arterial pressure and mean systemic arterial pressure (Pp/Ps). Aortic declamping was followed by a significant increase in lactate level and lactate-pyruvate ratio (L/P ratio) as compared with pre-declamping level ( P << 0.01). Although anaphylatoxin C3a concentration rose significantly after declamping ( P << 0.01), C5a showed no change at any stage. When the patients were divided into two groups according to the degree of Pp/Ps change by declamping, significant elevation of L/P ratio and C3a level were observed in the group with higher increase (Post-/Pre-declamp value >/=1.25) of Pp/Ps compared to the lower (>>1.25) group. After declamping, in the higher Pp/Ps group, a positive correlation existed not only between Pp/Ps change and aortic clamp time, but also between L/P ratio and C3a level. The present results suggest that muscular metabolic derangement distal to the aortic clamp may play an important role in the development of post-declamping pulmonary hypertensive response through anaphylatoxin C3a.

We have investigated the effects of anesthetic and related agents on Ca(2+)-induced Ca2+ release (CICR) in heavy sarcoplasmic reticulum isolated from rabbit skeletal muscle. The purpose of this study is to elucidate their possible role as triggering agents in malignant hyperthermia (MH). None of the agents (ketamine, procaine, lidocaine, succinylcholine, pancuronium and fentanyl) affected CICR at clinical concentrations. At higher concentrations, procaine, pancuronium and succinylcholine inhibited CICR, but ketamine rather potentiated it. It is unlikely that lidocaine is a potent facilitator of CICR at any concentrations. We conclude that procaine, lidocaine, non-depolarizing muscle relaxants and opiate can be used safely for MH susceptible patients and that ketamine and succinylcholine are not recommended.

Kinetic and thermodynamic parameters were evaluated for the acylation and the deacylation steps in the hydrolysis of p-nitrophenyl acetate by alpha-chymotrypsin at pH 7.8 and at temperatures between 15 and 35 degrees C by the use of stopped-flow and ordinary ultraviolet spectrophotometers. In contrast to the temperature dependencies of k2 and Ks reported in the literature (P.A. Adams and E.R. Swart, Biochem. J., 161, 83 (1977], no kinetic anomaly was observed in either of the steps, but reasonable straight lines were obtained in both Arrhenius and van't Hoff plots. On the other hand, in the chymotryptic hydrolysis of N-benzoyl-L-alanine methyl ester a sharp kinetic anomaly was found. The discrepancy in the case of p-nitrophenyl acetate is discussed in connection with a possible conformational change of the enzyme, an alteration of the rate-limiting step or differences in the experimental procedures. The cause of the anomaly observed in the case of N-benzoyl-L-alanine methyl ester is also discussed in detail.

The effects of succinylcholine and calcium (Ca2+), alone and together, on membrane-bound acetylcholinesterase ("true-type" cholinesterase) were examined using human erythrocyte ghosts to elucidate the combined pharmacological activity of succinylcholine and calcium in in vivo system. Succinylcholine inhibited the acetylcholinesterase by a mixed style. Calcium alone exhibited an inhibitory effect on the enzyme, but a biphasic effect together with succinylcholine: marked restoration of the enzyme activity at calcium concentrations lower than 6 mM and depression at its higher concentrations. It is suggested that calcium induces a conformational change of the enzyme protein leading to the altered binding capacity of succinylcholine. In anesthetic practice, therefore, the use of calcium may not be indicated for the treatment of SCh phase II block.