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We describe the identification and expression cloning of two novel enzymes, a beta-glucanase and an aspartic protease, secreted from the basidiomycetous yeast Phaffia rhodozyma. A cDNA library from P. rhodozyma CBS 6938 was constructed, and full-length cDNA encoding an endo-1,3(4)-beta-glucanase (bg1) and an aspartic protease (pr1) were cloned by expression cloning in Saccharomyces cerevisiae W3124. The bg1 cDNA encodes a 424-residue precursor protein with a putative signal peptide. The pr1 cDNA encodes a 405-residue prepropolypeptide with an 81-residue leader peptide. The aspartic protease was purified and characterized. It has a molecular mass of 36 kDa, an isoelectric point of pH 7.5, a pH activity optimum at 4.0-6.0, and a temperature activity optimum around 40 degrees C. Both enzymes show only low sequence identity to other known enzymes.

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The aim of this study was to describe the changes in frequency and character of interpersonal violence in Aarhus during a twelve year period. A one year (1993-1994) prospective registration of all persons exposed to violence and attending the casualty wards or the Department of Forensic Medicine was performed. The results were compared with similar studies performed in 1981-1982 and 1987-1988. The incidence in the last one year period had decreased significantly to 5.5 cases/1000 inhabitants compared to 6.5 in 1981-1982 and 7.5 in 1987-1988. The decrease was most distinct among men between 15-24 years of age. Ninety percent of the lesions were minor and the severity did not differ from the former study. Among risk factors associated to severe violence were men between 25-49 years of age, foreigners, use of knives and domestic violence. Firearms were only involved in 0.8% of the cases. Significantly higher proportion of the victims reported the incidence to the police in 1993-1994 than previously.

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In the period 1972-1992, 26 women had surgery for congenital vaginal agenesis, with a modified McIndoe's technique, in Aarhus University Hospital. Their medical records were reviewed and 23 of the patients were evaluated clinically. Complications comprised lack of skin graft take, bleeding, urethrovaginal fistula, perforation of the rectum, rectovaginal fistula and vaginal stricture. All complications were treated and the final results were satisfactory, i.e. complete graft take, satisfactory dimensions of the vagina and no strictures or fistulas giving symptoms. Of 23 patients reviewed, 18 reported satisfactory sexual relationships and vaginal intercourse was anatomically possible in a further 3 patients. We conclude that the McIndoe procedure is simple and, though the course of treatment is often protracted, serious complications are few and the final results are good.

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From 1970 to 1986 a total of 4578 hip fractures were registered in persons > or = 40 years in a Danish urban municipality. During that period, the annual number almost tripled. The statistical analysis revealed an increase in the age-specific incidence for both females and males (p < 0.0001). Furthermore, the number of fractures increased due to demographic aging. The overall incidence for individuals > or = 40 years increased 9-10% annually; from 2.1 to 5.4 per 1.000 females, and from 0.9 to 2.1 per 1.000 males. The female:male ratio was 2.8 without changes over time. If the observed increase in incidence is extrapolated to the next 17 years, a doubling of the annual number of fractures is predicted by the year 2002, whereas an unchanged incidence would result in a 20% increase in number.

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A Danish investigation of skiing injuries is presented from the casualty wards of two hospitals. The material consisted of 119 skiing injuries. Fifty-three per cent of the patients were women. Eighty-three injuries occurred abroad, mainly in Austria and Norway. Injury rate was not related to sex. The highest injury rate was found in the age group 20 to 29 years old. The most frequent injuries were sprains (knee 43, finger 14), fractures (39) and contusions (19). Injuries requiring hospitalization were common. None of these resulted in permanent major disability.

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The TM1 yeast mutant was transformed with a 2 micron-derived plasmid (YEp24) which carries a chimaeric gene containing the Escherichia coli chloramphenicol acetyl transferase (CAT) gene fused to the 5'- and 3'-flanking regions of the soybean leghemoglobin (Lb) c3 gene. Expression of the chimaeric CAT gene is controlled specifically by heme at a post-transcriptional level, most likely by regulating the efficiencies of translation. Expression of another chimaeric gene consisting of the neomycin phosphotransferase (NPTII) gene fused to only the 5'-flanking region of the Lbc3 gene is regulated by heme in a similar way. Thus, in yeast, heme modulates the translation of the chimaeric mRNAs through interactions with the 5' Lbc3 non-coding region.

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A DNase, designated Exonuclease II, has been purified 2,000-fold from whole cell extracts of bakers' yeast. It degrades single-stranded but not double-stranded DNA. The enzyme has a pH optimum around 8 and requires at least 2 mM Mg2+ for activity. It is slightly stimulated by dithiothreitol and inhibited by N-ethylmaleimide, suggesting that --SH groups are essential for activity; heparin also inhibits the activity. With 0.1 enzyme unit the Km has been determined to 4 nM of DNA ends and Vmax to 0.52 pmol of liberated nucleotides per min. The Mr is around 120,000. The enzyme does not degrade circular single-stranded M13 DNA, whereas linearized M13 DNA is degraded. The products are 5'-deoxyribomononucleotides exclusively. Using 5'-end labeled and 3'-end labeled DNA fragments as substrates, partially degraded DNA is only detectable in the latter case, showing that the exonuclease solely attacks DNA from the 5'-end. This distinguishes Exonuclease II from other exonucleases degrading DNA from the 5'-end, since they all either produce a mixture of 5'-mono- and oligonucleotides or 3'-mononucleotides. Analysis of 3'-end labeled fragments after incubation shows that the rate of exonucleolytic cleavage was dependent on the DNA sequence at the 5'-end.

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The ribonucleoside triphosphate, deoxyribonucleoside triphosphate, 3' -diphosphate guanosine 5' -diphosphate (ppGpp), and 5-phosphoribosyl 1-pyrophosphate (PRPP) pools in Escherichia coli B were determined by thin-layer chromatography during changing conditions to ammonium starvation. The intracellular concentrations of all nucleotides were found to change in a well-defined order several minutes before andy observed change in the optical density of the culture. The levels of purine nucleoside triphosphates (adenosine 5' -triphosphate [CTP], dCTP) and uridine nucleotides (uridine 5' -triphosphate, deoxythymidine 5'-triphosphate). The deoxyribonucleotides thus behaved as the ribonucleotides. The levels of ppGpp increased 11-fold after the decrease in uridine nucleotides, when the accumulation of stable ribonucleic acid (RNA) stopped. The level of the nucleotide pool did not stabilize until 30 min after the change in optical density. The pool of dGTP dropped concomitantly with the pool of CTP. The nucleotide precursor PRPP exhibited a transient increase, wtih maximum value of four times the exponential levels at the onset of starvation. Apparently the cell adjusts early to starvation by reducing either the phosphorylating activity or the nucleotide biosynthetic activity. As in other downshift systems, the accumulation of stable RNA stopped before the break in optical density and before the stop in protein accumulation. Cell divisions were quite insensitive to the control mechanisms operating on RNA and protein accumulation under ammonium starvation, since the cells continued to divide for 21 min without any net accumulation of RNA.

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