[Changing pattern of the respiratory function associated with resected lung lobe].

Respiratory function before and 2 months after lung lobectomy was analyzed associated with resected lobe. Post- or preoperative ratios of FEV1.0 or VC were compared among (1) predicted value by the number of subsegments using bronchofiberscopy, (2) predicted value by the lobar volume ratio using computed tomography (CT), and (3) actually measured value. Using subsegments method, post- or preoperative predicted VC ratios were 85 +/- 1% after right upper lobectomy (RU), 69 +/- 1% after right lower lobetomy (RL), 74 +/- 1% after left upper lobectomy (LU), and 75 +/- 1% after left lower lobectomy (LL). Using CT method, post- or preoperative predicted VC ratios were 80 +/- 2% after RU, 76 +/- 4% after RL, 74 +/- 2% after LU, and 79 +/- 3% after LL. Actually measured post- or preoperative FEV1.0 ratios were 82 +/- 3% after RU, 89 +/- 8% after RL, 73 +/- 3% after LU, and 86 +/- 5% after LL, and the VC ratios were 88 +/- 5% after RU, 79 +/- 3% after RL, 77 +/- 4% after LU, and 94 +/- 3% after LL. In the FEV1.0 analysis using both subsegments method and CT method, the predicted value was correlated with upper lobectomy but was overestimated in case of lower lobectomy. This phenomenon might be caused by the postoperative bronchial branching deformity after upper lobectomy. In the VC analysis using subsegments method, the predicted value was correlated with upper lobectomy but was overestimated in case of lower lobectomy. Meanwhile, in the VC analysis using CT method, the predicted value was correlated with RL or LU but was overestimated in case of RU or LL. This may due to the fact that RL and LU had large lobar volumes. In conclusion, postoperative predicted and actually measured values were different associated with resected lobe. In the FEV1.0 and VC analysis using subsegments method, the predicted value was strongly correlated with upper lobectomy but was overestimated (10%) in case of lower lobectomy.

[New subjects for exceeding conventional on-pump CABG].

Coronary artery bypass grafting (CABG) used to be performed under cardiac arrest and cardiopulmonary bypass (CPB). During the last decade, efforts were made to minimize CPB-related complications. The technique of off-pump CABG (OPCAB) has been established during the last 5 years. Elimination of CPB and OPCAB has successfully reduced a number of perioperative complications and has provided early patient recovery. A compression type of coronary stabilizer was used early phase of OPCAB. Off-pump revascularization using the compression device was limited to the anterior wall of the heart. Bypass to the posterior wall under a beating heart was not popular until the suction type of stabilizer had become available. A suction device assisted by the Lima's pericardial suture allowed us to perform bypass grafting any aspects of the heart. Recently, we are skeltonizing the arterial grafts using the Harmonic scalpel. Applying skeltonizing technique to the radial artery or internal thoracic artery, we can successfully perform sequential grafting in selected cases. The number of distal anastomoses has been gradually increased as the device and technique were advanced (2.1 distal anastomoses with a compression device, 2.9 with a suction device, and 3.2 with the skeltonization technique). The frequency of the complete revascularization also increased. On the other hand, the complications associated with the procedure were comparable among these three off-pump methods, but were significantly fewer than on-pump CABG. Currently performed OPCAB can provide almost same number of distal anastomoses as on-pump CABG, with less frequency of postoperative mortality and morbidity, and with early patient recovery. These favorable results were attributed to the progress of the device and technique.

Experimental myocardial preservation study of adding perfluorochemicals (FC43) in lidocaine cardioplegia.

OBJECTIVE: Lidocaine exhibits a cardioplegic action via acute inhibition of sodium influx into the myocardial cells. In terms of the cardiac function and calcium dynamics in the myocardial cells, we investigated the myocardial protective effect of addition of FC43 of Perfluorochemicals, which has an excellent oxygen transport function to meet the myocardial oxygen demand, on lidocaine-induced cardioplegia. METHODS: Isolated rat hearts were perfused with Langendorff mode and were divided to three experimental groups. During of preservation, these hearts were perfused continuously with the next three solution, potassium chloride was added to Krebs-Henseleit bicarbonate buffer to make potassium concentration of 20 mM in the first group (Group A), 2 mM lidocaine was added to Krebs-Henseleit bicarbonate buffer in the second group (Group B), and 2 mM lidocaine and 20% FC43 were added to Krebs-Henseleit bicarbonate buffer in the third group (Group C). After 60 minutes of continuous perfusion, the cardiac function and the intracellular calcium concentration in Groups A and B during cardioplegia were measured. Furthermore, after 360 minutes of continuous coronary perfusion, the cardiac function were measured in Group B and Group C. RESULTS AND CONCLUSIONS: Lidocaine cardioplegia showed a good recovery of cardiac function, because lidocaine induced prompt cardiac arrest by blocking sodium influx and inhibited the intracellular calcium overload by the following inhibition of sodium-calcium channels. Moreover, our results suggested that combining Perfluorochemicals with lidocaine produced a more effective myocardial-preservation that meets the myocardial oxygen demand during long-term cardiac arrest.

Heterologous expression, purification, and kinetic comparison of the cytoplasmic and mitochondrial glyoxalase II enzymes, Glo2p and Glo4p, from Saccharomyces cerevisiae.

The aims of the present study are (i) to purify a mitochondrial glyoxalase II to homogeneity for the first time from any organism and (ii) to compare its kinetic properties with those of the cytoplasmic enzyme. Both the cytoplasmic and the mitochondrial glyoxalases II from Saccharomyces cerevisiae, which are the products of two distinct genes, GLO2 and GLO4, were purified from yeast and in recombinant form from Escherichia coli. To obtain a higher protein yield (compared to wild-type expression) in yeast, the genes were placed under the control of the strong GAL1 promoter on a multicopy plasmid. Amino-terminal sequencing and molecular mass determination by MALDI-TOF mass spectrometry of the mitochondrial Glo4 protein revealed Met-11 of the primary translation product of the gene as the N-terminal amino acid. Judged by enzyme kinetic properties the recombinant and natural proteins were equivalent. The cytoplasmic and the mitochondrial enzyme differed in the pH dependence of the kinetic parameters for the main substrate, S-d-lactoylglutathione. Whereas the cytoplasmic protein showed a pronounced peak of enzyme activity between pH 7-8 and a continuous up to fivefold increase of the K(M) value with increasing pH (from 5. 5-9.0), the mitochondrial protein had a nearly constant K(M) value and an activity maximum over a broad pH range (6.5-9.0). The kinetic parameters (at pH 7.5) of both the cytoplasmic and the mitochondrial enzyme for S-D-lactoylglutathione were of the same order of magnitude as reported recently for the human and Arabidopsis thaliana enzymes which are presumably of cytoplasmic origin. However, both yeast enzymes showed a severalfold lower preference for the more hydrophobic substrate, S-d-mandeloylglutathione.

Identification and phenotypic analysis of two glyoxalase II encoding genes from Saccharomyces cerevisiae, GLO2 and GLO4, and intracellular localization of the corresponding proteins.

We have isolated and characterized two genes coding for the glyoxalase II enzyme from Saccharomyces cerevisiae. The coding region of the GLO2 gene corresponds to a protein with 274 amino acids and a molecular mass of 31,306 daltons. The open reading frame of the GLO4 gene could be translated into a protein with 285 amino acids and a molecular mass of 32,325 daltons. The amino acid sequences of the deduced proteins are 59.1% identical and show high similarities to the sequence of the human glyoxalase II. When grown on either glucose or glycerol as a carbon source, a glo2 glo4 double deletion strain contains no glyoxalase II activity at all and shows no obvious phenotype during vegetative growth. However, this strain showed a similar high sensitivity against exogenous methylglyoxal as compared with a glyoxalase I-deficient strain. Whereas the GLO2 gene is expressed on both glucose and glycerol, the GLO4 gene is only active on glycerol. The active Glo2p protein is localized in the cytoplasm and the active Glo4p in the mitochondrial matrix. Heterologous expression of the full-length GLO2 coding region in Escherichia coli resulted in an active protein. However, to get an active Glo4p protein in E. coli, the putative mitochondrial transit peptide at the N-terminal end had to be removed by shortening the 5' end of the GLO4 open reading frame.

Revised analysis of aadA2 gene of plasmid pSa.

The nucleotide sequence of gene aadA2 of plasmid pSa, coding for aminoglycoside-3"-adenyltransferase, has been reexamined. We found differences with respect to the sequence previously determined by Tait et al. (R. C. Tait, H. Rempel, R. L. Rodriguez, and C. I. Kado, Gene 36:97-104, 1985). These deviations are located in the coding region and in the 3' noncoding region. By making deletions in the region for initiation of protein synthesis, we identified a GTG triplet as the most probable start codon for translation.

The gene coding for the major birch pollen allergen Betv1, is highly homologous to a pea disease resistance response gene.

Pollen of the white birch (Betula verrucosa) is one of the main causes of Type I allergic reactions (allergic rhinoconjunctivitis, allergic bronchial asthma) in Middle and Northern Europe, North America and the USSR. Type I allergies are a major threat to public health in these countries, since 10-15% of the population suffer from these diseases. BetvI, an allergenic protein with an Mr of 17 kd is a constituent of the pollen of white birch and is responsible for IgE binding in more than 95% of birch pollen allergic patients. Here, we report the complete nucleotide sequence and deduced amino acid sequence of a cDNA clone coding for the major pollen allergen (BetvI) of white birch. It is similar to the N-terminal peptide sequences of the allergens of hazel, alder and hornbeam (close relatives) but it has no significant sequence homology to any other known allergens. However, it shows 55% sequence identity with a pea disease resistance response gene, indicating that BetvI may be involved in pathogen resistance of pollen.