Cloning of genes encoding polyketide synthases (PKSs) has allowed us to identify a gene cluster for ML-236B biosynthesis in Penicillium citrinum. Like lovastatin, which is produced by Aspergillus terreus, ML-236B (compactin) inhibits the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. Genomic sequencing and Northern analysis showed that nine predicted genes for ML-236B biosynthesis were located within a 38-kb region and were transcribed when ML-236B was produced. The predicted amino acid sequences encoded by these nine genes, designated mlcA- mlcH and mlcR, were similar to those encoded by the genes for lovastatin synthesis, and were therefore assumed to be involved either directly or indirectly in ML-236B biosynthesis. Targeted disruption experiments provided evidence that two PKS genes in the cluster, mlcA and mlcB, are required for the biosynthesis of the nonaketide and the diketide moieties, respectively, of ML-236B, suggesting that the gene cluster as a whole is responsible for ML-236B biosynthesis in P. citrinum. Bioconversion of some of the predicted intermediates by an mlcA-disrupted mutant was also investigated in order to analyze the ML-236B biosynthetic pathway. The molecular organization of the gene cluster and proposed functions for the ML-236B biosynthetic genes in P. citrinum are described.

OVERVIEW OF THE LARGE HELICAL DEVICE PROJECT. The Large Helical Device (LHD) has successfully started running plasma confinement experiments after a long construction period of eight years. During the construction and machine commissioning phases, a variety of milestones were attained in fusion engineering which successfully led to the first operation, and the first plasma was ignited on 31 March 1998. Two experimental campaigns are planned in 1998. In the first campaign, the magnetic flux mapping clearly demonstrated a nested structure of magnetic surfaces. The first plasma experiments were conducted with second harmonic 84 and 82.6 GHz ECH at a heating power input of 0.35 MW. The magnetic field was set at 1.5 T in these campaigns so as to accumulate operational experience with the superconducting coils. In the second campaign, auxiliary heating with NBI at 3 MW has been carried out. Averaged electron densities of up to 6 × 10 19 m-3 , central temperatures ranging from 1.4 IAEA-F1-CN-69/OV1/4 2 to 1.5 keV and stored energies of up to 0.22 MJ have been attained despite the fact that the impurity level has not yet been minimized. The obtained scarling of energy confinement time has been found to be consistent with the ISS95 scaling law with some enhancement.

High-spatial-resolution multichannel Thomson scattering measurement scheme is applied to the diagnostics of a microplasma produced between two knife-edge electrodes separated by a 100 µm microgap and driven by 2.45 GHz microwave. The method provides spatial profile of electron density, electron temperature, and gas temperature simultaneously in a single measurement, with a spatial resolution of ∼25 µm; the gas temperature is derived from the spectrum of rotational Raman scattering caused by molecules contained in the discharge gas. Measurements are carried out for discharges with air and He/N2(5%) mixture gas at atmospheric pressure and the differences in the basic plasma parameters are discussed.

To determine whether elderly patients have an increased incidence of postoperative pneumonia and/or an increased mortality, we conducted a retrospective study of 4,380 patients who underwent thoracic, abdominal or neurosurgery under general anesthesia. Thirty of these patients developed postoperative pneumonia (incidence, 0.68%). The elderly patients aged 65 years or older who underwent thoracic surgery had the highest incidence of pneumonia. Nine of the 30 patients with pneumonia died (mortality, 30%). Elderly patients did not show a higher mortality than the younger patients. The extent of pulmonary infiltrates on chest roentgenogram and the serum bilirubin level, both of which were determined at the onset of pneumonia, independently had a significant association with mortality. Our study suggests that advanced age is a risk factor for postoperative pneumoniain patients whoundergo thoracic surgery. (Internal Medicine 32: 274-277, 1993)

The gene cluster responsible for ML-236B (compactin) biosynthesis has recently been characterized from P. citrinum No. 41520. Here, we describe how the ML-236B-producing strain was improved using a cosmid-mediated recombination technique. The introduction of the cosmid pML48, which contains seven of the nine ML-236B biosynthetic genes, into P. citrinum No. 41520 resulted in transformants which produced increased amounts of ML-236B. Southern analysis showed that pML48 had been incorporated by a homologous recombination event, and all high producers possessed two copies of each of the seven genes, mlcA- mlcF and mlcR, suggesting that increased dosage of the biosynthetic gene cluster was responsible for the enhanced production of ML-236B. On the other hand, various kinds of mutants with decreased titers of ML-236B were also obtained. Characterization of one such mutant, designated as T48.28, which was more sensitive to ML-236B than the parental strain, suggested that one of the ML-236B biosynthetic genes, mlcD, which encodes a putative HMG-CoA reductase, was involved in conferring resistance to ML-236B.