Legionella gresilensis sp. nov. and Legionella beliardensis sp. nov., isolated from water in France.

Novel Legionella-like isolates, strains Montbéliard A1T and Gréoux 11 D13T, isolated from two different French water sources, were studied taxonomically and phylogenetically. Morphological and biochemical characterization revealed that they were Gram-negative, aerobic, non-spore-forming bacilli with a cut-glass appearance that grew only on L-cysteine-supplemented buffered charcoal yeast extract agar. Phenotypic characterization using fatty acid and ubiquinone profiles and SDS-PAGE analysis confirmed that they were closely related, but distinct from, other species of the genus Legionella, since serotyping could not relate them to any existing serogroup. Genotypic profiles generated by randomly amplified polymorphic DNA and 16S-23S rDNA spacer region PCR analyses were unique for each of these isolates. DNA-DNA relatedness values of strains Montbéliard A1T and Gréoux 11 D13T to each other and to other Legionella type strains were less than 25%. Phylogenetic affiliation of these organisms obtained by 16S rDNA sequence comparisons confirmed that they were distinct from any other known Legionella species. All the above results confirm that these strains constitute two novel species for which the names Legionella gresilensis sp. nov. (type strain Gréoux 11 D13T = ATCC 700509T = CIP 106631T) and Legionella beliardensis sp. nov. (type strain Montbéliard A1T = ATCC 700512T = CIP 106632T) are proposed.

Transrectal high-intensity focused ultrasound: minimally invasive therapy of localized prostate cancer.

BACKGROUND AND PURPOSE: Criteria for determining the durability of the response to transrectal high-intensity focused ultrasound (HIFU) ablation of prostate cancer have been established by calculating progression-free probability. PATIENTS AND METHODS: A series of 82 patients (mean age 71 +/- 5.7 years) with biopsy-proven localized (stage T1-T2) cancer who were not suitable candidates for radical surgery underwent transfectal HIFU ablation with the Ablatherm machine. The mean follow-up was 17.6 months (range 3-68 months). The mean serum prostate specific antigen (PSA) value and mean prostate volume were 8.11 +/- 4.64 ng/mL and 34.9 +/- 17.4 cm3, respectively. Progression was rigidly defined as any positive biopsy result, regardless of PSA concentration, or three successive PSA increases for patients with a negative biopsy (PSA velocity > or = 0.75). Times to specific events (positive biopsy and PSA elevation) were analyzed with the Kaplan-Meier survival method. RESULTS: Overall, 62% of the patients exhibited no evidence of disease progression 60 months after transrectal HIFU ablation. In particular, the disease-free rate was 68% for the moderate-risk group of 50 patients (PSA < 15.0 ng/mL, Gleason sum < 8, prostate volume < 40 cm3, and number of positive biopsies < 5). For the low-risk group of 32 patients (PSA < 10 ng/mL and Gleason sum < 7), the disease-free survival rate was 83%. CONCLUSION: Transrectal HIFU prostate ablation is an effective therapeutic alternative for patients with localized prostatic adenocarcinoma.

Staphylococcus fleurettii sp. nov., isolated from goat's milk cheeses.

A new coagulase-negative and novobiocin-resistant species of the genus Staphylococcus, Staphylococcus fleurettii, isolated from raw-milk cheeses, is described. This species is differentiated from the other novobiocin-resistant staphylococci on the basis of ribotype and intergenic transcribed spacer patterns, DNA-DNA reassociation reactions, cell wall composition and phenotypic characteristics. S. fleurettii could be distinguished by its oxidase activity, by its ability to produce acid aerobically from D-trehalose, D-mannose, D-turanose and maltose and by its inability to produce acid from D-cellobiose. The type strain of S. fleurettii is CIP 106114T (= DSM 13212T).

Peritoneal clearance of leptin in continuous ambulatory peritoneal dialysis.

Leptin is a 16-kd protein that increases energy expenditure and limits food intake. Serum leptin (S-leptin) is elevated in dialysis patients, and little data have been reported on leptin clearance (Cl) during dialysis. We analyzed the peritoneal dialysis (PD) Cl of leptin in 15 continuous ambulatory peritoneal dialysis (CAPD) patients and compared the results to beta(2)-microglobulin (beta(2)-m), urea, and creatinine PD Cl. S-leptin was significantly elevated (Kruskal-Wallis, P < 0.005) in CAPD women (58.4 +/- 42.4 [SE] microg/L, n = 5) as compared with CAPD men (13.9 +/- 7.1, n = 10) and with healthy women (11.0 +/- 1.4, n = 13) and men (5.1 +/- 0. 9, n = 14). Correlations were found between percent of fat mass and S-leptin (P < 0.05); between S-leptin and the 24-hour PD leptin (P < 0.05); and between dialysate-to-plasma (D/P) beta(2)-m and D/P leptin (P < 0.01). PD leptin Cl (1.80 +/- 0.43 mL/min/1.73 m(2)) was higher than beta(2)-m Cl (1.22 +/- 0.31) (P < 0.01), but reduced as compared with urea Cl (8.84 +/- 1.20) (P < 0.005) and creatinine Cl (7.71 +/- 0.99) (P < 0.005). These results indicate that leptin is eliminated through the peritoneum membrane. However, peritoneal leptin clearance, as beta(2)-m, appears to be clearly restricted as compared with peritoneal transport of smaller molecules. Hence, leptin could use the same diffusion transport pathway as beta(2)-m. In addition, leptin, which has a higher molecular weight than beta(2)-m, was significantly more eliminated into the peritoneal dialysate. More studies are necessary to clarify whether this is an active leptin elimination process by peritoneal secretion or by a different restriction coefficient of diffusion through the peritoneum membrane.

Differential RNA expression of the pS2 gene in the human benign and malignant prostatic tissue.

PURPOSE: The pS2 trefoil protein has been detected in close association with neuro-endocrine differentiation in prostate cancer and prostatic intraepithelial neoplasia. These preliminary results have suggested that pS2 is a candidate as a specific marker for prostate cancer tissue. To ascertain the specificity of pS2 in prostate cancer tissue, we have used an RT-PCR method from prostate biopsies provided from human malignant and benign prostatic hyperplasia (BPH) tissue. MATERIALS AND METHODS: Prostate biopsies were obtained from transrectal biopsies from 153 patients with an abnormal DRE or a PSA more than 4 ng./ml. or symptoms of BPH and a PSA more than 4 ng./ml. Total RNA was extracted from fresh frozen specimens of tissue samples. Detection of pS2 transcript compared with GADPH transcripts was done using RT-PCR. RESULTS: Biopsy results showed that 108 patients had prostate cancer (average Gleason score 6.39+/-0.74) and 45 patients had BPH. PS2 RT-PCR results showed that PS2 RNA expression was negative in 83% of the BPH cases. Conversely, 92% of prostate cancer specimens were positive (Chi-square: 86.09, p<0.001). There was no correlation with tumor stage or the Gleason score. Comparing the expression of pS2 in BPH and localized prostate cancer, we found a sensitivity of 92% and a specificity of 82%. CONCLUSIONS: On this large sample of prostate biopsies from patients at risk of having prostate cancer, pS2 was demonstrated as an interesting marker significantly associated with prostate cancer. Further work on the expression of pS2 according to differentiation and hormonal status is in progress.

Legionella taurinensis sp. nov., a new species antigenically similar to Legionella spiritensis.

A group of 42 Legionella-like organisms reacting specifically with Legionella spiritensis serogroup 1 antisera were collected throughout Europe by the Centre National de Référence (French National Reference Centre) for Legionella. This group of isolates differed somewhat from L. spiritensis in terms of biochemical reactions, ubiquinone content and protein profile. The latter two analyses revealed that one of these L. spiritensis-like isolates, Turin I no. 1T, was highly related, but not identical to any of the red autofluorescent species of Legionella. In fact, this strain was the first of these particular isolates recognized to emit a red autofluorescence when exposed to UV light. Profile analysis of randomly amplified polymorphic DNA established that the red autofluorescent L. spiritensis-like isolates constituted a homogeneous group distinct from Legionella rubrilucens and Legionella erythra. DNA-DNA hybridization studies involving the use of S1 nuclease confirmed that the indicated group of isolates are a new species of Legionella, for which the name Legionella taurinensis is proposed with strain Turin I no. 1T (deposited as ATCC 700508T) as the type strain.

Local control of prostate cancer by transrectal high intensity focused ultrasound therapy: preliminary results.

PURPOSE: We conducted a phase I/II study to evaluate the efficacy of transrectal high intensity focused ultrasound in the treatment of localized prostate cancer and to assess associated complications. The efficacy of a new high intensity ultrasound device was evaluated using post-treatment prostate specific antigen (PSA) levels and histological results from prostate biopsies as end points. MATERIALS AND METHODS: A total of 113 transrectal high intensity focused ultrasound sessions were performed in 50 patients with localized prostate cancer, who were not suitable candidates for radical prostatectomy. Of these patients 2 underwent salvage ultrasound treatment for locally recurrent cancer following definitive radiation therapy. Mean plus or minus standard deviation patient age, PSA and prostate volume were 70.7+/-4.54 years, 9.61+/-7.42 ng./ml. and 37.3+/-19.1 cc. The 2 different high intensity ultrasound prototypes were successfully used, and the latter prototype included several safety devices to reduce morbidity. Median followup was 24 months (range 3 to 46). Control parameters were changes in PSA and random control sextant biopsies at 1 to 3, 3 to 12, 12 to 24, 24 to 36 and 36 to 48 months. RESULTS: For the evaluation of therapy patients were divided into 4 groups. Group 1 (complete response) included 28 patients (56%) with no residual cancer and PSA less than 4 ng./ml. (mean 0.93), group 2 (biochemical failure) 3 patients (6%) with no residual cancer and PSA greater than 4 ng./ml. (mean 6.22), group 3 (biochemical control) 9 patients (18%) with residual cancer (mean positive biopsy 1.1 of 6) and PSA less than 4 ng./ml. (mean 0.90), and group 4 (failures) 10 patients (20%) with residual cancer (mean positive biopsies 1.9 of 6) and PSA greater than 4 ng./ml. (mean 8.9). Of the 10 cases in group 4 hormone therapy was required in 3 and radiotherapy in 5. Complication rate with the first prototype device was 50% and it decreased to 17% with the second prototype. CONCLUSIONS: Morbidity associated with high intensity focused ultrasound treatment is currently minimal. Local control of the localized prostate cancer was observed in groups 1, 2 and 3 (80%). Repeat sessions were deferred in groups 2 and 3 based on changes in PSA. These preliminary data suggest that high intensity focused ultrasound represents a valid alternative treatment strategy for patients with localized prostate cancer who are unsuitable for surgery.

Acute leptin regulation in end-stage renal failure: the role of growth hormone and IGF-1.

BACKGROUND: Leptin, a recently discovered peptide involved in nutrient intake and energy expenditure, has been shown to be abnormally regulated in certain conditions such as obesity. In chronic renal failure, leptin appears to be increased. However, little is known about leptin regulation during chronic renal failure (CRF). METHODS: We measured serum leptin in eight well nourished, chronic hemodialysis patients (seven males, one female) receiving anabolic factors for three days as either recombinant insulin-like growth factor-1 (rhIGF-1) or a combination of recombinant growth hormone (rhGH) plus recombinant IGF-1, in a random cross-over trial. RESULTS: Serum leptin values were in the range of normal volunteers matched for body mass index. As reported in other conditions, serum leptin was strongly correlated with patients dry body wt (P = 0.01) and body fat (P = 0.0001). Both treatments affected serum leptin in a rapid and opposite manner. RhIGF-1 decreased serum leptin from 11.2+/-20.8 (SD) to 4.3+/-3.8 microg/liter (P = 0.011), whereas the combination of rhGH + rhIGF-1 increased serum leptin from 7.4+/-9.4 to 21.0+/-32.9 microg/liter (P = 0.011). Regression analyses indicated a linear regression between serum leptin and insulin variations after treatment. CONCLUSIONS: This study shows for the first time that both rhIGF-1 and rhGH acutely regulate serum leptin in dialysis patients. Whether leptin changes are explained by the concomitant insulin variation should be further studied under renal failure conditions.

Non-linear patterns of growth in very low birth weight infants.

The deviations of daily weight, weekly length and head circumference from linear growth were analyzed in 87 very low birth weight infants. The deviations exhibited a highly significant polynomial oscillation not only for weight, but also for length and head circumference. The weight amplitudes were larger for males than for females. They were also larger in infants appropriate for gestational age than in small-for-date infants. The difference with trophicity may be related to the process of adjustment of catch-up in small-for-date infants. However, the mechanisms of these oscillatory deviations could also be explained by clinical events, method of feeding, or homeostatic regulation. Further studies are required to elucidate the role of the different factors. Am. J. Hum. Biol. 10:637-646, 1998. © 1998 Wiley-Liss, Inc.

Involvement of DNA methylation in the control of the expression of an estrogen-induced breast-cancer-associated protein (pS2) in human breast cancers.

pS2 gene has been used to investigate the relationship between alterations of DNA methylation patterns in human tumors and gene expression. The expression of pS2, which is transcriptionally controlled by estrogens in breast cancer cell lines, is restricted to estrogen-receptor-rich human breast tumors. We found that the CCGG site within the promoter/enhancer sequence of pS2 was hypomethylated in estrogen-receptor-rich breast tumors expressing this gene. The amount of DNA molecules unmethylated at this site was related to the amount of pS2 mRNA detected in the samples. The demethylation of this region, which contains the estrogen responsive element, was confirmed by genomic sequencing. Transient expression of functional human estrogen receptors stimulated the expression of the endogenous pS2 in HeLa cells, but failed, in BT-20 cells, to stimulate expression of this gene. Since the promoter/enhancer region of pS2 is unmethylated in HeLa cells and methylated in BT-20 cells, these data also support the hypothesis that DNA methylation might be involved in the control of pS2 expression.

Identification of atypical strains of Staphylococcus epidermidis by use of molecular tools.

Four atypical coagulase-negative staphylococcal (CNS) isolates from clinical sources were compared with Staphylococcus epidermidis strains by ribotyping. The ribotypes of the four strains shared close rDNA restriction profiles with those of the S. epidermidis strains used. The DNA sequence encoding 16S rRNA demonstrated 99.9% homology with S. epidermidis. S1 nuclease experiments showed that these atypical strains formed a homogeneous genomic group. DNA-DNA homologies between the S. epidermidis type strain CCM 2124 and the four CNS isolates ranged from 70 to 89%. The guanine-plus-cytosine content of the deoxyribonucleic acid of the four strains ranged from 31 to 32 mol%.

Staphylococcus saprophyticus subsp. bovis subsp. nov., isolated from bovine nostrils.

A new coagulase-negative subspecies, Staphylococcus saprophyticus subsp. bovis, is described on the basis of a study of five strains isolated from the anterior nares of cows. This subspecies is differentiated from the other novobiocin-resistant staphylococci by its phenotypic properties, cell wall composition, and levels of genetic relatedness. The type strain of the new subspecies is KV 12 (=CCM 4410).

Comparison of six serum ferritin immunoassays and isoferritin spectrotypes in malignancies.

Serum ferritin was measured by six enzyme immunoassays in specimens from patients with digestive cancers (n = 30) and hematologic malignancies (n = 33). Most mean comparisons show significant differences in both groups of patients. In digestive cancers correlations between any two methods are very satisfactory (r > 0.99) but a proportional bias is often observed. In hematologic malignancies, correlations are bad (r < 0.80 in 8 out of 15 correlations) because of many discrepant values. Isoelectric focusing separation of isoferritins was performed in most specimens and the pattern of each serum was compared to the between kit CV. We conclude that an 'acid' spectrotype increases between-kit analytical variability. We try to explain the results taking into account the nature of the immunological systems and the cross-reactions with tissular isoferritins. In conclusion, our results indicate that large differences may be observed in sera from hematologic malignancies (leukemias, lymphomas ... ) We recommend that monitoring be achieved by the same method of measurement.

Expression of an estrogen-induced breast cancer-associated protein (pS2) in benign and malignant human ovarian cysts.

BACKGROUND: In human mammary tumors, pS2 expression is directly controlled by estrogens and restricted to a subclass of breast carcinomas. In addition, recent studies have suggested that this gene is expressed in both the invasive and preinvasive forms of breast cancer. EXPERIMENTAL DESIGN: pS2 gene expression was investigated in benign and malignant ovary tumors and whenever possible, pS2 expression was also studied in cells collected from cystadenoma fluids. In several cases, particularly with cells from cystadenoma fluids, the limited amount of material available prevented the used of the traditional RNA detection methods such slot/dot blots or Northern blots. Therefore, a rapid and sensitive polymerase chain reaction assay of pS2 expression has been developed and used in this study. RESULTS: In human ovarian tumors, data obtained show that pS2 transcripts and proteins are present in all mucinous cystadenomas studied and at a lower frequency in endometrioid cystadenomas. Quantitation of the CA 19-9 mucin concentration in ovarian fluids indicate that pS2 expression is always associated with high mucin concentrations, but mucin-positive and pS2-negative samples are also frequently observed. CONCLUSIONS: These data suggest that pS2 expression is restricted to subclasses of human ovarian cystadenomas.

[Osteocalcin and smoking]. / Ostéocalcine et tabagisme.

Smoking is a risk factor for osteoporosis. Nicotine and nonnicotine tobacco smoke components have been shown to depress osteoblast activity in a number of in vitro and animal studies. To determine whether smoking is associated with depressed osteoblast activity in humans, we measured serum osteocalcin levels (using a radioimmunological method based on an antibody to human osteocalcin) in 24 male or female smokers and 24 matched nonsmokers. Overall, osteocalcin levels were significantly lower in smokers (15 +/- 6.95 ng/ml) than in nonsmokers (21.27 +/- 8.34 ng/ml) (p = 0.007). The difference between smokers and nonsmokers was significant in males (15.3 +/- 4.5 vs 23.27 +/- 9.7; p = 0.02) but not in females (16.27 +/- 8.9 vs 19.45 +/- 6.7; p = 0.2). These data suggest that smoking may induce osteoblast depression, either directly or via hormonal changes.

Serum ferritin and isoferritins are tools for diagnosis of active adult Still's disease.

OBJECTIVE: Still's disease is an acute systemic inflammatory disorder. There are no pathognomonic symptoms or specific laboratory abnormalities. Serum ferritin concentration in rheumatoid arthritis together with some plasma glycoproteins such as alpha 2-glycoprotein and C-reactive protein are part of the response to inflammation. Ferritin in plasma is glycosylated and the sialoglycosylated forms increase its microheterogeneity. Our purpose was to confirm in a large series that high values of ferritin can be found in adult Still's disease (ASD) and to see if a specific isoferritin can be isolated in this disease compared with the other systemic diseases. METHOD: Thirty-one sera were investigated from 11 men and 9 women with ASD and compared with 27 sera from 27 patients with systemic diseases. We studied the course of one case of ASD for 15 months. Serum ferritin was determined by immunoenzymology (Abbott Ferrizin). The isoferritins were investigated by isoelectric focussing and the percentage of glycosylation by affinity for concanavalin A (Con-A). RESULTS: In patients with active ASD, the ferritin levels were higher than in patients with inactive ASD or other systemic diseases: p < 0.001. The glycoforms of ferritin were basic and the proportion of ferritin bound to Con-A was lower than other ASD: p < 0.001. CONCLUSIONS: Serum ferritin levels have a diagnostic value for acute ASD. The study of sialylation and abnormalities in the glycosylation of ferritin helps to discriminate ASD from arthritis or other systemic diseases. In conclusion, the glycoform of isoferritins and the percentage of glycosylation offers an additional tool for the diagnosis of Still's disease.

Specific binding between human neutrophils and heparin.

Heparin binding on polymorphonuclear leucocytes (PMNL) was characterized. Heparin binding was specific, rapid, saturable and reversible. One single class of heparin binding sites was found with a dissociation constant of 1.22 mumol/l and 7.7 x 10(6) sites per PMNL. The binding was independent of the anticoagulant activity of heparin. Heparin affinity chromatography on radio-iodinated cell lysates followed by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate revealed a 130 kD heparin binding protein. Heparin binding was inhibited by disodium salt of ethylenediamine tetraacetic acid. Cell surface bound heparin was functionally inactive and did not affect the inactivation of thrombin by antithrombin III. Our study demonstrates that heparin interacts with PMNL by a cell-surface binding protein. These instructions could be consistent with the modifications of some PMNL functional properties in the presence of heparin.

[Echographic guided puncture of ovarian cysts. Possibilities and limitations]. / Ponction échoguidée des kystes de l'ovaire. Possibilités et limites.

Ultrasound-guided puncture is a simple and easy to perform procedure. It would seem to be a good idea to suggest simple puncture as a first intention in cases of an image of ovarian cyst. In theory, the advantages are obvious: a puncture is performed, the liquid is analyzed and an appropriate treatment is administered. Coelio-surgery could surely be avoided in cases of functional cysts and perhaps in some non-malignant ovarian cysts. In fact, it must be remembered that a cancer of the ovary in its early stages may have the appearance of a banal cyst, and that puncture does not allow pathological examination. Cytological examination is insufficient to totally rule out malignancy or to allow detailed histological diagnosis. There is, therefore, a risk of leaving in place the pocket of a cyst which may be organic and which may recur or even develop. For these reasons, ultrasound-guided puncture can be undertaken only in pre-selected patients and in the context of a specific protocol: 1) The ultrasound image of the cyst must be liquid, anechoic, unilocular (or bilocular with a fin wall), with no vegetation, the serum level of CA 125 must be low; 2) it the puncture liquid is oily, tarry or viscous, a celioscopy must be carried out as soon as possible, only a yellow-colored liquid can justify waiting; 3) the analysis of the cyst fluid is not always determinant, and the cytology findings are conclusive only if positive. A high 17 beta-estradiol level suggests a functional cyst.(ABSTRACT TRUNCATED AT 250 WORDS)