Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440.

Pseudomonas putida is a metabolically versatile saprophytic soil bacterium that has been certified as a biosafety host for the cloning of foreign genes. The bacterium also has considerable potential for biotechnological applications. Sequence analysis of the 6.18 Mb genome of strain KT2440 reveals diverse transport and metabolic systems. Although there is a high level of genome conservation with the pathogenic Pseudomonad Pseudomonas aeruginosa (85% of the predicted coding regions are shared), key virulence factors including exotoxin A and type III secretion systems are absent. Analysis of the genome gives insight into the non-pathogenic nature of P. putida and points to potential new applications in agriculture, biocatalysis, bioremediation and bioplastic production.

Progress in Arabidopsis genome sequencing and functional genomics.

Arabidopsis thaliana has a relatively small genome of approximately 130 Mb containing about 10% repetitive DNA. Genome sequencing studies reveal a gene-rich genome, predicted to contain approximately 25000 genes spaced on average every 4.5 kb. Between 10 to 20% of the predicted genes occur as clusters of related genes, indicating that local sequence duplication and subsequent divergence generates a significant proportion of gene families. In addition to gene families, repetitive sequences comprise individual and small clusters of two to three retroelements and other classes of smaller repeats. The clustering of highly repetitive elements is a striking feature of the A. thaliana genome emerging from sequence and other analyses.

[Geriatric health system, economic profit and the debate on euthanasia]. / Geriatrische Versorgung, ökonomischer Nutzen und die Euthanasiedebatte.

Three characteristic developments in modern western societies usually are considered to be independent variables in the ethical discussion:1. An explosion-like increase in medical and social expenditures following a rapid multiplication of old and multiply disabled people in this century. 2. the increasing economic importance of the "health industry", and 3. the "new" debate of euthanasia. All these developments are discussed controversially. The volume of geriatric support is mostly considered to be insufficient, but usually it is restricted by both, scarcity of resources as well as increasing demands to focus on "evidence-based medicine" (which might exclude a lot of medical procedures in old age). The mutation of health systems from - originally - social activities to business branches more and more gives priority to economical based decisions in medicine, but otherwise has advanced an increasing number of new health professions. The origin of the actual debate on euthanasia is the development and judicial certification of individual's self-determination in modern societies. However, euthanasia is still refused world-wide because it is considered to be linked with a process of weakening basic ethical principles. The 3 seemingly independent developments certainly are facts in modern societies. They hardly can be influenced by the medical profession, being forced to conform to them. However, there are significant connections between them.A geriatric health system, primarily denying individual demands and basic convictions of old people, contributes to an attitude of non-acceptance towards daily practice medicine. The same effect may result from the economic transformation of medicine when creating a system of self-perpetuating demand (being characteristic for an "ideal" business branch) by "unlimited" prolongation of life of the very old and highly disabled patients. The result from this development undoubtedly will be an increasing demand for self-determination at the end of life, including medical assistance in suicide and euthanasia, which cannot successfully be confronted with moral appeals.Alternatively, a basic correction of the geriatric health system must be introduced mainly including psychodynamic factors in medical decision making, or euthanasia will be accepted by the majority, as already has happened in the Netherlands.

Identification of the essential EPE1 gene involved in retention of secreted proteins on the cell surface of Saccharomyces cerevisiae cells.

Saccharomyces cerevisiae yeast cells secrete extracellularly low amounts of a few proteins. The reasons for retardation of secreted proteins on the cell surface remain obscure. We describe here a mutant able to export enhanced amount of proteins. Classical genetic methods, nucleic acids manipulations and cloning procedures were used to isolate and characterize the mutant and to clone and sequence the corresponding wild type gene. The isolated Saccharomyces cerevisiae mutant MW11, is temperature sensitive and exports on average twenty-fold more proteins at 37 degrees C than parental wild type strain (80 micrograms of proteins/1 x 10(8) mutant cells, SEM +/- 5, n22; versus 3 micrograms of proteins/1 x 10(8) parental cells, SEM +/- 1, n22). Protein overexport in the mutant requires a functional SEC1 pathway and is independent of cell lysis. Cloning and sequencing of the corresponding wild type gene identified an open reading frame of 786 bp coding for a hydrophilic protein with predicted molecular mass of 30 kDa and cytosolic localization. The newly identified gene, designated EPE1, is an essential gene. Its DNA and amino acids sequence showed no homology with other yeast genes and proteins. It is concluded that the function of unknown yet genes, such as EPE1 is needed for retention of secreted proteins on the surface of Saccharomyces cerevisiae cells.

Sequence and analysis of chromosome 4 of the plant Arabidopsis thaliana.

The higher plant Arabidopsis thaliana (Arabidopsis) is an important model for identifying plant genes and determining their function. To assist biological investigations and to define chromosome structure, a coordinated effort to sequence the Arabidopsis genome was initiated in late 1996. Here we report one of the first milestones of this project, the sequence of chromosome 4. Analysis of 17.38 megabases of unique sequence, representing about 17% of the genome, reveals 3,744 protein coding genes, 81 transfer RNAs and numerous repeat elements. Heterochromatic regions surrounding the putative centromere, which has not yet been completely sequenced, are characterized by an increased frequency of a variety of repeats, new repeats, reduced recombination, lowered gene density and lowered gene expression. Roughly 60% of the predicted protein-coding genes have been functionally characterized on the basis of their homology to known genes. Many genes encode predicted proteins that are homologous to human and Caenorhabditis elegans proteins.

Renal abscesses in an AIDS patient caused by Aspergillus fumigatus.

BACKGROUND: Aspergillus abscesses of the kidneys are very rare complications of AIDS which usually have fatal outcome. The 4 patients described in the literature, 2 of whom had bilateral involvement, died due to this opportunistic infection. METHODS: Case report of a 33-year-old patient with AIDS in stage C3 and aspergillus abscesses of the kidney. RESULTS: Successful treatment of bilateral aspergillus abscesses of the kidneys with purely conservative antimycotic measures. This was largely due to stabilization of the immune status by supplementing antiretroviral treatment with the proteinase inhibitor indinavir. CONCLUSIONS: The options now available for improving immune status in AIDS also improve the treatment chances in renal aspergillosis.

Analysis of 1.9 Mb of contiguous sequence from chromosome 4 of Arabidopsis thaliana.

The plant Arabidopsis thaliana (Arabidopsis) has become an important model species for the study of many aspects of plant biology. The relatively small size of the nuclear genome and the availability of extensive physical maps of the five chromosomes provide a feasible basis for initiating sequencing of the five chromosomes. The YAC (yeast artificial chromosome)-based physical map of chromosome 4 was used to construct a sequence-ready map of cosmid and BAC (bacterial artificial chromosome) clones covering a 1.9-megabase (Mb) contiguous region, and the sequence of this region is reported here. Analysis of the sequence revealed an average gene density of one gene every 4.8 kilobases (kb), and 54% of the predicted genes had significant similarity to known genes. Other interesting features were found, such as the sequence of a disease-resistance gene locus, the distribution of retroelements, the frequent occurrence of clustered gene families, and the sequence of several classes of genes not previously encountered in plants.

A temperature-sensitive lambda cI repressor functions on a modified operator in yeast cells by masking the TATA element.

We describe the construction and analysis of derivatives of the yeast TDH3 promoter in which the TATA box element has been replaced by a portion of the phage lambda operator containing a consensus TATA site flanked by binding sites for the cI repressor. Transcription of a reporter gene under the control of such a promoter is reduced in cells that express the cI repressor protein. Deletion of the native TATA element of the TDH3 promoter reduces transcription to the same extent. The cI repressor may act by "masking" the TATA element located between the repressor binding sites. Furthermore, the use of a temperature-sensitive cI repressor allowed temperature-dependent transcription of the reporter gene.

Chromosome polymorphisms close to the cm-ADE1 locus of candida maltosa.

The imperfect yeast Candida maltosa has an ill-defined genetic constitution; it is nominally diploid, but probably highly aneuploid, in nature. We report on polymorphisms specifically affecting those chromosomes which bear the cm-ADE1 gene. This gene encodes phosphoribosylaminoimidazole-succino-carboxamide synthetase, an enzyme in the adenine biosynthetic pathway. By electrophoretic karyotype analysis, three differently sized chromosomes were demonstrated to carry cm-ADE1; the size (but not the number) of these chromosomes was also found to vary, both between strains and during the mitotic growth of a single strain. Four different alleles of cm-ADE1 have been cloned and sequenced from one prototrophic strain. DNA sequence divergence between these different alleles is as high as 8%, with the greatest divergence being found in the upstream region. Mitotic recombination events that led to changes in the karyotype were followed by using cm-ADE1 DNA as an hybridization probe. A recombination hot-spot in the neighbourhood of the gene appears to be responsible for the instability of the chromosomes on which it residues.

IASP task force on euthanasia and assisted suicide.

The first meeting of the IASP Task Force on Euthanasia and Assisted Suicide took place in Venice on June 7, 1995. Several interested observers were present. It was decided that at the public IASP meeting the following day each speaker should address, briefly, the current legal situation and the pressure for change, as well as give a personal statement. David Clark spoke for North America, Bob Goldney for Australia, Michael Kelleher for Britain and Ireland, Jerzy Wasserman for Scandinavia, and Hans Wedler for the German-speaking world. Their views are published in this article. Ad Kerkhof requested that the Dutch television film "Death on Request" be discussed. The committee was of the opinion that clear definitions were essential. In their view, these should take into account the differences between active and passive euthanasia, as well as between professionally assisted and lay-assisted suicide.

Efficient electropulse transformation of intact Candida maltosa cells by different homologous vector plasmids.

Conditions for efficient and quick transformation by electroporation were developed in Candida maltosa. To investigate the efficiency of transformation with integrative as well as with autonomously replicating plasmids, a series of vectors was constructed for homologous transformation of this species. Transformants were obtained with different plasmids as covalently closed circular molecules and as linearized DNA. The influence of recipient strain and plasmid type as well as of cell number and parameters of the supplied electrical pulse on the transformation efficiency have been investigated. A maximum of 7000 transformants per 100 ng of plasmid DNA was reached. The efficiency of transformation was compared with that of the LiCl method.

Correlation of biochemical blocks and genetic lesions in leucine auxotrophic strains of the imperfect yeast Candida maltosa.

The four enzymatic steps in the conversion of alpha-ketoisovaleriate to leucine were examined in the wild type and in 13 leucine auxotrophic strains of Candida maltosa. The genetic lesions in the auxotrophs, involve at least five different loci and are correlated with three enzymatic steps. This was confirmed by gene cloning, protoplast fusion, and enzyme assays. The pathway for leucine biosynthesis in C. maltosa shows general similarity to that of other lower eukaryotes but there are individual differences in the numbers of genes responsible for single enzymatic steps. A disomic state of the chromosomes carrying genes coding for alpha-isopropylmalate synthase and beta-isopropyl-malate dehydrogenase was elucidated.

[The concept of integrated internal medicine-psychosomatic patient management--experiences and results with its application in a general hospital]. / Das Konzept der integrierten internistisch-psychosomatischen Patientenversorgung--Erfahrungen und Ergebnisse bei der Umsetzung im Allgemeinen Krankenhaus.

Psychosomatic therapy has been realized by integration into general ward's therapeutic program in a general hospital. Within 3 1/2 years of development the medical staff has been qualified to perform basic elements of psychosomatic therapy, whilst the structure of the medical department has been changed in different aspects. During a period of 28 days the real extend of psychosomatic therapy now has been evaluated by questionnaire: 438 psychosomatic and psychosocial interventions were documented in this time, concerning 149 patients (= 53% of all patients treated in the department). Causes, trigger mechanic, settings, frequency, median duration, and different kinds of all these interventions are described. According to the clientèle of our medical department, with a lot of elderly and multi-handicapped persons, the main topics of interventions concerned--besides of specific psychosomatic treatment--addiction and problems of alcoholism, coping strategies and compliance, as well as further support of handicapped singles after discharge.