Analysis of six prophages in Lactococcus lactis IL1403: different genetic structure of temperate and virulent phage populations.

We report the genetic organisation of six prophages present in the genome of Lactococcus lactis IL1403. The three larger prophages (36-42 kb), belong to the already described P335 group of temperate phages, whereas the three smaller ones (13-15 kb) are most probably satellites relying on helper phage(s) for multiplication. These data give a new insight into the genetic structure of lactococcal phage populations. P335 temperate phages have variable genomes, sharing homology over only 10-33% of their length. In contrast, virulent phages have highly similar genomes sharing homology over >90% of their length. Further analysis of genetic structure in all known groups of phages active on other bacterial hosts such as Escherichia coli, Bacillus subtilis, MYCOBACTERIUM: and Streptococcus thermophilus confirmed the existence of two types of genetic structure related to the phage way of life. This might reflect different intensities of horizontal DNA exchange: low among purely virulent phages and high among temperate phages and their lytic homologues. We suggest that the constraints on genetic exchange among purely virulent phages reflect their optimal genetic organisation, adapted to a more specialised and extreme form of parasitism than temperate/lytic phages.

Nucleotide sequence of the putative regulatory gene and major promoter region of the Streptomyces griseus glycerol operon.

Nucleotide sequencing of the deduced major promoter region of the glycerol utilization operon and an upstream regulatory gene of Streptomyces griseus reveals extensive similarity to the previously sequenced homologous S. coelicolor region [Smith and Chater, J. Mol. Biol. 204 (1988) 569-580]. However, regions showing extensive divergence are found in the noncoding parts of the sequence. These may help to evaluate the significance of various sequence features in relation to promoter activity.

Cloning, sequence and expression in Escherichia coli of the Methylobacillus flagellatum recA gene.

By means of interspecific complementation of an Escherichia coli recA- mutation with phasmids containing a gene bank from an obligate methylotroph, Methylobacillus flagellatum (Mf), the recA+ gene from this bacterium was identified. When expressed in an E. coli recA- host, it can function in recombination, DNA repair, and prophage induction. The nucleotide sequence of the gene has been determined. The coding region consists of 1032 bp specifying 344 amino acids. The deduced RecA protein structure shows a striking homology with RecA from other bacteria, except for the C-terminal region and some residues which were proposed to be responsible for the coprotease ability of RecA proteins. The region preceding the recA-Mf gene start codon has no SOS box--the LexA repressor binding site. Expression of the recA-Mf gene in E. coli proved to be DNA-damage independent.

Construction and transduction of a shuttle vector bearing the cos site of Streptomyces phage phi C31 and determination of its cohesive ends.

A plasmid has been constructed which is a bifunctional vector for Escherichia coli and for streptomycetes, containing the cos site of Streptomyces phage phi C31. It can be efficiently transduced into S. lividans 66 and into several other Streptomyces species. The nucleotide sequence of a 311-bp DNA fragment containing the cohesive ends has been determined. The cohesive ends consist of 10 bases protruding at the 3'-end of the phage genome.

Living with a child with familial Mediterranean fever: does it affect the quality of life of the parents?

OBJECTIVE: The aim of the present study was to assess the quality of life (QOL) and the psychological status of parents of children with familial Mediterranean fever (FMF). METHODS: The QOL, anxiety and depression of the parents of 35 children with FMF were evaluated and compared to the parents of 23 healthy children. RESULTS: Mothers of FMF children had lower QOL scores than mothers of healthy children: 5.5 +/- 1.1 versus 6.0 +/- 0.6 (p = 0.048). They also expressed higher levels of anxiety and depression. Within each group, mothers were more anxious and depressed than fathers. Parents with several FMF children were not significantly different from parents with only one FMF child. CONCLUSION: The QOL and psychological well being of parents with FMF children were found to be slightly impaired, especially that of the mothers.

Assessment of quality of life of parents of children with juvenile chronic arthritis.

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low.

The effect of balneotherapy at the Dead Sea on the quality of life of patients with fibromyalgia syndrome.

Fibromyalgia (FS) is an idiopathic chronic pain syndrome defined by widespread non-articular musculoskeletal pain and generalised tender points. As there is no effective treatment, patients with this condition have impaired quality of life (QoL). The aim of this study was to assess the possible effect of balneotherapy at the Dead Sea area on the QoL of patients with FS. Forty-eight subjects participated in the study; half of them received balneotherapy, and half did not. Their QoL (using SF-36), psychological well-being and FS-related symptoms were assessed prior to arrival at the spa hotel in the Dead Sea area, at the end of the 10-day stay, and 1 and 3 months later. A significant improvement was reported on most subscales of the SF-36 and on most symptoms. The improvement in physical aspects of QoL lasted usually 3 months, but on psychological measures the improvement was shorter. Subjects in the balneotherapy group reported higher and longer-lasting improvement than subjects in the control group. In conclusion, staying at the Dead Sea spa, in addition to balneotherapy, can transiently improve the QoL of patients with FS. Other studies with longer follow-up are needed to support our findings.

CLINHAQ scale--validation of a Hebrew version in patients with fibromyalgia. Clinical Health Assessment Questionnaire.

Assessment of health status in patients with rheumatic disease, including fibromyalgia (FM), using structured questionnaires has become an important approach to evaluate treatment and outcome. The objectives of this study were to validate a translated version of the Clinical Health Assessment Questionnaire (CLINHAQ) to be used by Hebrew-speaking populations, and specifically to evaluate its usefulness in fibromyalgia syndrome (FM). The CLINHAQ was translated into Hebrew and administered to 90 women with FM along with the Hebrew versions of the Fibromyalgia Impact Questionnaire (FIQ) and the Quality of Life (QOL) Scale. The CLINHAQ includes scales of functional disability, helplessness, anxiety and depression, as well as assessment of current health status and satisfaction with this. All subjects were asked about the presence and severity (assessed by visual analogue scale) of current FM symptoms (pain, fatigue, anxiety etc.); a count of 18 tender points was conducted by thumb palpation, and tenderness thresholds were measured by dolorimetry. Test-retest reliability was assessed by Pearson correlation coefficients, and internal consistency was evaluated with Cronbach's alpha coefficient of reliability. Construct validity was tested by correlating the CLINHAQ items with measures of symptom severity, count of tender point, tenderness thresholds, physical functioning measured by FIQ, and with a score of QOL. Test-retest reliability coefficients ranged from 0.82 to 0.99, and Cronbach's alpha coefficients from 0.725 to 0.929. Significant moderate to high correlations were obtained between most subscales of CLINHAQ and measures of physical functioning, quality of life and severity of FM symptoms. In conclusion, the CLINHAQ is a reliable and valid instrument for measuring health status and physical functioning in Israeli women with FM.

Dietary and clastogenic factors in children who immigrated to Israel from regions contaminated by the Chernobyl accident.

The authors evaluated the possible association between dietary history and plasma clastogenic factors in children who immigrated to Israel between 1989 and 1993 from regions contaminated by the Chernobyl accident. The authors compared questionnaire data about demographic variables, dietary histories before and after immigration occurred, and health status with clastogenic factor scores for 162 immigrants. Logistic regression analysis revealed a negative association between clastogenic factor scores and frequency of consumption of fresh vegetables and fruit among children < or = 7 yr of age during the postimmigration period. Intake of eggs and fish by boys who were < or = 7 yr of age prior to immigration was associated positively with clastogenic factor scores. Consumption of fresh vegetables and fruits afforded protection to the immune systems of children who were < or = 7 yr of age.

[Structure of the Bacillus brevis metalloprotease gene]. / Struktura gena metalloproteazy Bacillus brevis.

Primary structure of DNA fragment of 2355 b.p., encoding metalloprotease gene of Bac. brevis, had been determined. Open reading frame for a protein with size of 528 amino acid residues was found in this sequence. The encoding protein is homologous to metalloproteases of Bac. stearothermophilus, Bac. cereus, Bac. subtilis and Bac. amyloliquefaciens. The structure of Bac. brevis metalloprotease gene reveals that this enzyme is synthesised as pre-pro-protease with signal peptide and pro-region, which are cut during its synthesis. The proposed size of mature protease is 304 amino acid residues. The residues, essential for catalysis, binding of Zn ion and Ca ions were found on the basis of Bacilli metalloproteases structures comparison.

[Cloning and expression in Bacillus subtilis of the gene for neutral protease of Bacillus brevis]. / Klonirovanie i ékspressiia v Bacillus subtilis gena neitral'noi proteazy Bacillus brevis.

Plasmids pCB20 and pCB22 were used for cloning and expression of the Bac brevis 7882 neutral protease gene in Bac. subtilis cells. The protease-containing fragments of 13 and 14 kb were cloned in pCB20 plasmid based on replication region of Streptococci plasmid pSM19035. Expression of the gene was shown to take place in Bac. subtilis. Application of vegetative promoters of the previously identified expression unit EU19035 greatly increases the expression of the protease in Bac. subtilis. Bac. subtilis cells, expressing the gene of Bac. brevis neutral protease, do not sporulate, are considerably larger than the cells which do not contain the gene and form multicellular structures.

[Site-specific restrictases from Bacillus thuringiensis var. Kumantoensis]. / Saitspetsificheskie restriktazy iz shtamma Bacillus thuringiensis var.Kumantoensis.

The efficiency of bacteriophages CP-54 and CP-55 plating on Bacillus thuringiensis var. kumantoensis H18 (Kum) is decreased about 10-fold as compared with the efficiency of plating on Bacillus thuringiensis var. galleriae H5 (Gal). Bacteriophages having propagated for one cycle in Kum cells might be further grown in this strain without growth restriction. Two site-specific restriction enzymes isolated from Bacillus thuringiensis var. kumantoensis were designated BtkI and BtkII. The endonuclease BtkI recognises the same nucleotide sequence CGCG in DNA as recognised by the restriction endonuclease FnuDII; BtkII recognises the same nucleotide sequence GATC as the endonuclease Sau3A.

[A new site-specific endodeoxyribonuclease EcoHI]. / Novaia saitspetsificheskaia éndodezoksiribonukleaza EcoHI.

A new sitespecific endonuclease of the II class EcoHI has been isolated from Escherichia coli strain and characterized. Restriction endonuclease EcoHI recognises the nucleotide sequence C C (C/G) G G with the cleavage site between the fourth and fifth nucleotide. It is an isoshizomer of the restriction endonuclease CauII. The yield of enzyme is 2500 units of activity per 1 g of biomass. The producing strain Escherichia coli HI is nonpathogenic, easily grown with the antibiotic resistance markers permitting to cultivate the strain under selective conditions.

[Intergeneric Escherichia coli-Streptomyces conjugation as a means for the transfer of conjugative plasmids into producers of antibiotics chlortetracycline and bialaphos]. / Mezhrodovaia kon"iugatsiia Escherichia coli-Streptomyces kak sposob peredachi kon"iugativnykh plazmid v produtsenty antibiotikov khlortetratsiklina i bialafosa.

A system for introduction of plasmids into industrial producers of antibiotics chlortetracycline and bialaphos using intergeneric conjugation of Escherichia coli and Streptomyces was developed. Low level stability of inheritance of autonomously replicating DNA in recipient strains was shown. Site-specific integration of the conjugative-integrative vector pTO1 provided stable plasmid maintenance within the chromosomes of Streptomyces aureofaciens and S. hygroscopicus. Phenomenon of disturbance in differentiation and antibiotic production, resulting from pTO1 integration into S. hygroscopicus chromosome, was discovered.

[Streptavidin secretion by Streptomyces lividans strains]. / Sekretsiia streptavidina shtammov Streptomyces lividans.

Recombinant strains of S. lividans capable of secreting streptavidin were isolated. Various constructions containing either streptavidin gene copies integrated within a chromosome or a streptavidin gene within the secretory vector were investigated. S. lividans SA2 containing a gene copy integrated within the chromosome had the maximum productivity amounting to 60-80 per cent of the S. avidinii productivity. New vectors for Streptomyces used in the study are described.

Probiotics improve bowel movements in hospitalized elderly patients--the PROAGE study.

OBJECTIVE: To determine the impact of probiotics on the prevention of problems with bowel movements malnutrition and infection. DESIGN: A randomized, double-blind, placebo-controlled trial. SETTING: Peripheral Geriatric Hospital. PARTICIPANTS: 243 elderly patients age ≥ 65 y who were hospitalized in a Geriatric Orthopedic Rehabilitation Department. INTERVENTION: Participants were randomized into treatment or control groups (daily probiotics or placebo for 45 consecutive days, respectively). MEASUREMENTS: The main outcomes were: number of days of constipation or diarrhea and the number of days of laxative use. Secondary measures were nutritional status and blood measurements. RESULTS: Of 599 patients admitted to the Geriatric Rehabilitation ward, 345 were eligible and agreed to participate. During a 7-day pre-trial period, 102 patients dropped out (45 and 57 in the probiotic and placebo groups respectively). Out of the 243 patients who entered the study, 28 dropped out during the study (11.5%), leaving 215 patients. Throughout the 45 days of follow-up, the incidence of diarrhea was significantly lower among the study group (HR=0.42, p=0.04) with a more pronounced difference among participants aged ≥ 80 y (HR=0.32, p=0.026). Laxative use (as an indicator of constipation severity) was significantly lower in the study group compared with the control group (HR=0.74, p=0.032). Serum albumin, prealbumin and protein increased significantly more in the treatment group compared with the control group among participants age ≥ 80 y (P=0.047, p=0.07, p=0.03 respectively) but not in the younger age group. CONCLUSION: We showed that probiotic supplements may have a positive effect on bowel movements among orthopedic rehabilitation elderly patients.

Sense of coherence and its impact on quality of life of patients with systemic lupus erythematosus.

The sense of coherence (SOC) construct refers to a global orientation, which significantly determines the link between stressors, coping with disease and health. The aim of this work was to assess possible associations between SOC and quality of life (QOL) scores among women with SLE. Sixty consecutive SLE women and 88 healthy women were included in the study. QOL was assessed using the SF-36 and the WHO QOL-Bref scales. The SOC has three main sub-scales: comprehensibility, manageability and meaningfulness. Regression analyses were used to study associations between various parameters of SF-36, WHO QOL-Bref, SOC, SLEDAI, indices of end organ damage (SDI), and demographic variables. Mean SLEDAI and SDI scores were 4.5 (SD = 5.6) and 1.29 (SD=2). SLE patients had significantly lower scores for all individual and summary sub-scales in the two QOL questionnaires compared with controls. SLE patients had significantly lower scores for the general, comprehensibility and meaningfulness sub-scales of SOC. No significant correlation was seen between SOC scores and measures of disease activity or end-organ damage. A strong linear correlation was seen between the scores of SOC, general WHO QOL-Bref, and the mental and physical component summary (MCS & PCS) scores of SF36. Age, SOC and SDI significantly affected the PCS score. SOC was the only variable independently associated with MCS. Education and SOC were significantly associated with the general WHO QOL-Bref. Age, education, SDI and SOC were independently associated with QOL of women with SLE.

The complete genome sequence of Lactobacillus bulgaricus reveals extensive and ongoing reductive evolution.

Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) is a representative of the group of lactic acid-producing bacteria, mainly known for its worldwide application in yogurt production. The genome sequence of this bacterium has been determined and shows the signs of ongoing specialization, with a substantial number of pseudogenes and incomplete metabolic pathways and relatively few regulatory functions. Several unique features of the L. bulgaricus genome support the hypothesis that the genome is in a phase of rapid evolution. (i) Exceptionally high numbers of rRNA and tRNA genes with regard to genome size may indicate that the L. bulgaricus genome has known a recent phase of important size reduction, in agreement with the observed high frequency of gene inactivation and elimination; (ii) a much higher GC content at codon position 3 than expected on the basis of the overall GC content suggests that the composition of the genome is evolving toward a higher GC content; and (iii) the presence of a 47.5-kbp inverted repeat in the replication termination region, an extremely rare feature in bacterial genomes, may be interpreted as a transient stage in genome evolution. The results indicate the adaptation of L. bulgaricus from a plant-associated habitat to the stable protein and lactose-rich milk environment through the loss of superfluous functions and protocooperation with Streptococcus thermophilus.

Fine particles and meteorological conditions are associated with lung function in children with asthma living near two power plants.

Fine particles are thought to pose a risk to health, especially for vulnerable groups such as children with asthma. These children are also known to be affected by meteorological and seasonal changes. We assessed the association between air pollution and lung function via peak expiratory flow (PEF), controlling for seasonal changes, meteorological conditions and personal physiological, clinical and sociodemographic measurements, in a panel of schoolchildren with asthma living near two power plants in Israel. Two hundred and eighty-five children with confirmed asthma performed PEF tests and completed a respiratory symptoms diary twice a day. Particulate matter <10 microm in diameter (PM10), particulate matter <2.5 microm in diameter (PM2.5) and meteorological conditions were measured at six fixed stations. Data were analysed using time series analysis-generalized linear model and generalized estimating equations. The models were built under the assumption that any health outcome belongs to a multivariate hierarchical system and depends on meteorological, geophysical and sociocultural variables and pollution factors. No significant differences were found in the demographic (age, gender, mean parental education level, parental smoking habits, place of birth and housing density), physiological (body mass index) and clinical factors (illness severity) between the communities participating in the study. A significant direct effect of PM2.5 on the PEF was found in Ashdod (P=0.000). In Sderot, this effect was through an interaction between PM10 and the sequential day of the year (P=0.000). The main conclusion of this study is that children with asthma are at risk from air pollution and geophysical conditions. Policy makers should take these results into consideration when setting thresholds for environmental protection.