Mycobacterium tuberculosis multi-drug-resistant strain M induces IL-17+ IFNγ- CD4+ T cell expansion through an IL-23 and TGF-ß-dependent mechanism in patients with MDR-TB tuberculosis.

We have reported previously that T cells from patients with multi-drug-resistant tuberculosis (MDR-TB) express high levels of interleukin (IL)-17 in response to the MDR strain M (Haarlem family) of Mycobacterium tuberculosis (M. tuberculosis). Herein, we explore the pathways involved in the induction of Th17 cells in MDR-TB patients and healthy tuberculin reactors [purified protein derivative healthy donors (PPD+ HD)] by the M strain and the laboratory strain H37Rv. Our results show that IL-1ß and IL-6 are crucial for the H37Rv and M-induced expansion of IL-17+ interferon (IFN)-γ- and IL-17+ IFN-γ+ in CD4+ T cells from MDR-TB and PPD+ HD. IL-23 plays an ambiguous role in T helper type 1 (Th1) and Th17 profiles: alone, IL-23 is responsible for M. tuberculosis-induced IL-17 and IFN-γ expression in CD4+ T cells from PPD+ HD whereas, together with transforming growth factor (TGF-ß), it promotes IL-17+ IFN-γ- expansion in MDR-TB. In fact, spontaneous and M. tuberculosis-induced TGF-ß secretion is increased in cells from MDR-TB, the M strain being the highest inducer. Interestingly, Toll-like receptor (TLR)-2 signalling mediates the expansion of IL-17+ IFN-γ- cells and the enhancement of latency-associated protein (LAP) expression in CD14+ and CD4+ T cells from MDR-TB, which suggests that the M strain promotes IL-17+ IFN-γ- T cells through a strong TLR-2-dependent TGF-ß production by antigen-presenting cells and CD4+ T cells. Finally, CD4+ T cells from MDR-TB patients infected with MDR Haarlem strains show higher IL-17+ IFN-γ- and lower IL-17+ IFN-γ+ levels than LAM-infected patients. The present findings deepen our understanding of the role of IL-17 in MDR-TB and highlight the influence of the genetic background of the infecting M. tuberculosis strain on the ex-vivo Th17 response.

Extraction and partial characterization of proteolytic activities from the cell surface of Lactobacillus helveticus Zuc2.

Proteolytic activities were extracted from a dairy Lactobacillus helveticus strain and partially characterized. A first cell envelope proteinase (CEP) was extracted using a high ionic strength buffer, both in the presence and in the absence of Ca2+. Moreover, cell treatment by 5 M LiCl allowed for the selective removal of the S-layer protein and CEP, suggesting an enzyme ionic linkage to the cell envelope similar to that observed for the Slayer structure. The enzyme specificity against alpha(s1)-CN (f1-23) showed unusual activity on the Lys3-His4 bond compared with other proteinases of the same species. A second proteinase appeared to be linked to the cell membrane because it was extractable only after membrane disgregation by detergents. Its specificity against CN fractions and alpha(s1)-CN (f1-23) was different from that of the first CEP; moreover, the measured activity was lower than that of CEP.

Antimicrobial compounds from Lactobacillus casei and Lactobacillus helveticus.

Three strains of Lactobacillus casei and one of Lactobacillus helveticus were examined for antagonistic activity toward twenty five indicator strains of different species. Under conditions eliminating the effects of organic acid and hydrogen peroxide, culture supernatants of all the Lactobacillus strains exhibited a wide spectrum of inhibitory activity toward microorganisms of different genera. The inhibitory compound secreted by one strain of L. casei was active against Clostridium tyrobutyricum. The active components were insensitive to proteolytic enzyme and heat treatment.

Intestinal surgery in patients with ovarian carcinoma.

Twenty patients affected with ovarian carcinoma underwent intestinal operations between 1979 and 1987 during the course of the disease. The patients were divided into two groups on the basis of the time and the aim of the adopted surgery. The first group had intestinal surgery during the initial laparotomy for debulking of the tumour and amelioration of bowel function, while the patients of the second group underwent re-exploration at different times from the first laparotomy for symptomatic intestinal involvement by tumour, with the aim of palliation for obstruction. In all the cases the intestinal obstruction, complete or incomplete, was caused by tumoral involvement of the loops histologically proven. The patients were affected with ovarian carcinoma at stage III (80%) and IV (20%) FIGO. The survival at the intestinal surgery, correlated to the FIGO stage and the kind of surgery performed are presented.

A mixture of mivacurium and rocuronium is comparable in clinical onset to succinylcholine.

STUDY OBJECTIVES: To compare the clinical onset and duration of a combination of mivacurium and rocuronium with succinylcholine, and to determine the efficacy of this mixture for rapid tracheal intubation. DESIGN: Observer-blind prospective study. SETTING: Teaching hospital. PATIENTS: 70 ASA status I and II patients having general anesthesia for elective surgery. MEASUREMENTS AND MAIN RESULTS: After induction of general anesthesia, patients randomly received succinylcholine 1.0 mg/kg, rocuronium 0.6 mg/kg, or a combination of rocuronium 0.6 mg/kg and mivacurium 0.15 mg/kg. Evoked muscular response at the adductor pollicis was measured by mechanomyography. The time from injection of muscle relaxant(s) to ablation of T1 (clinical onset) and recovery of T1 to 25% of control height (clinical duration) was recorded. Intubating conditions 45 seconds after administration of muscle relaxants were assessed. There was no significant difference in clinical onset time between succinylcholine (mean +/- SD, 47.4 +/- 6.5 seconds) and the combination of mivacurium-rocuronium (51.2 +/- 13.4 seconds). Intubating conditions with mivacurium-rocuronium were comparable to those of succinylcholine. The clinical duration of rocuronium 0.6 mg/kg (38.9 +/- 12.3 minutes) was prolonged by the addition of mivacurium (49.0 +/- 9.6 minutes). CONCLUSIONS: This combination of mivacurium and rocuronium is comparable to succinylcholine in both clinical onset time and quality of intubating conditions. When rapid onset of dense neuromuscular blockade and intermediate clinical duration is desirable, this mixture may be an acceptable alternative to succinylcholine.

Pain evaluation during carbon dioxide laser vaporization for cervical intraepithelial neoplasia: a randomized trial.

63 pts affected by CIN of various degrees were randomly divided into 3 groups in order to evaluate the pain experienced during laser vaporization of the lesion. All pts were premenopausal and ages ranged between 19 and 39 years. 21 pts received Naproxene Sodium (550 mg) 30 minutes before surgery; 21 pts received placebo and 21 pts received no drug. Laser vaporization was performed with a Coherent System 451 CO2 laser with a power setting of 28 W/cm2 and a spot size of 1.8 mm. The severity of pain was assessed by means of a Visual Analogue Scale. The mean VAS value was 19 for the group treated with Naproxene Sodium; the mean VAS value was 20 for the placebo group and 23 for the group which received no pre-operative drug. Analysis of data from the 3 groups showed no statistically significant difference. Analgesia or anaesthesia before laser surgery for CIN is not a necessity.

Drug resistance plasmids in Lactobacillus acidophilus and Lactobacillus reuteri.

Sixteen strains of Lactobacillus reuteri and 20 strains of Lactobacillus acidophilus were tested for resistance to 22 antibiotics by using commercially available sensitivity disks. Evidence suggesting linkage of these resistances to plasmids was obtained by "curing" experiments with acridine dyes and high growth temperatures. Examination of plasmid patterns of agarose gel electrophoresis provided further evidence of loss in plasmid DNA under curing conditions in some of the strains examined.

Partial characterization and plasmid linkage of a non-proteinaceous antimicrobial compound in a Lactobacillus casei strain of vegetable origin.

Lactobacillus casei IMPC LC34 of vegetable origin produces a non-proteinaceous inhibitory compound with a broad spectrum of activity towards Gram-positive and Gram-negative bacteria, including pathogens. The active substance, mainly produced in the stationary phase of growth, is insensitive to proteolytic enzymes, lipase and catalase, and is stable at 121 degrees C for 30 min. The inhibitory activity was detected either at 8 degrees C or at 37 degrees C. The active compound does not contain glucidic groups, is inactivated by Na-metaperiodate, and its molecular mass is between 2000 and 5000 Da. Plasmid curing experiments showed that both antimicrobial compound immunity and production determinants were encoded by an 8.8 kbp plasmid. The effectiveness of the active agent was verified on ready-to-use vegetables, using either the Lact. casei strain or its culture supernatant fluid as inoculant, compared with cured clone. The application potential of the Lact. casei strain or its culture supernatant fluid for assuring the microbiological safety of ready-to-use vegetables is discussed.

Inhibitory effect of selected lactic acid bacteria on microflora associated with ready-to-use vegetables.

The addition of selected lactic acid bacteria strains had a remarkable inhibitory effect on the growth dynamics of microflora associated with ready-to-use vegetables, during refrigerated storage. In particular, coliforms and enterococci were strongly reduced or eliminated from the products from the third day of storage. Lactobacillus casei strains proved more effective than pediococci. The use of lactic cultures able to produce bacteriocins and to grow at low temperatures could be a useful tool to preserve fresh vegetables and to ensure their microbiological safety.

Fast and slow milk-coagulating variants of Lactobacillus helveticus HLM 1.

Slow milk-coagulating variants were isolated from Lactobacillus helveticus HLM 1, a fast strain which coagulates milk in 16 h at 42 degrees C. Variants were isolated after subculturing in reconstituted skim milk or complex broth media. Analysis of plasmid content revealed that in slow variants a 3.5-megadalton plasmid was missing.

Study on crop adhesion genetic determinant in Lactobacillus reuteri.

The genetic determinant responsible for adherence in one strain of Lactobacillus reuteri isolated from chicken crop has been investigated. After curing experiments carried out on this strain, a plasmid-free derivative was obtained, resulting unchanged as regards adhesion ability and carbohydrate fermentation pattern, but showing a loss of antibiotic resistances. It is suggested a possible localization on chromosome of adhesion determinant.

Partial characterization of a bacteriocin produced by Lactobacillus helveticus.

AIMS: To investigate the antimicrobial activity of a strain of Lactobacillus helveticus. METHODS AND RESULTS: The culture supernatant fluid Lact. helveticus G51 showed antimicrobial activity against thermophilic strains of Lactobacillus. Purification of the active compound was achieved after gel filtration and ion exchange chromatography. As revealed by SDS-PAGE, active fractions were relatively homogeneous, showing a protein with a molecular mass of 12.5 kDa. The antimicrobial compound was heat labile, inactivated by proteolytic enzymes and had a bactericidal mode of action. CONCLUSION: The antimicrobial activity expressed by Lact. helveticus G51 was correlated with the production of a bacteriocin with properties that were different to other helveticins. SIGNIFICANCE AND IMPACT OF THE STUDY: The study has provided further data on Lact. helveticus bacteriocins. The strong activity of the bacteriocin towards various thermophilic lactobacilli warrants further investigation for its potential to obtain attenuated cultures for the enhancement of the cheese-ripening process.

DNA-free liposome-assisted transfection of Streptococcus faecium SF 77R protoplasts.

Two virulent phages of Streptococcus faecium SF 77R were used in DNA-free liposome assisted polyethylene-glycol-mediated protoplast transfection. This procedure, established to develop an effective gene-cloning system, optimized the conditions for DNA uptake by the protoplasts of Str. faecium. Transfection frequencies of up to 10(7)-10(8) P.F.U./microgram of DNA were achieved.

Antagonism and adhesion among isogenic strains of Lactobacillus reuteri in the caecum of gnotobiotic mice.

Interactions between a wild adhesive Lactobacillus reuteri strain isolated from chicken crop and its isogenic plasmid-free derivative were studied. Antagonism between these strains was tested in vitro or associated with gnotobiotic mice. The cured strains repressed the establishment of the wild, plasmid bearing strain in the gastrointestinal tract of gnotobiotic mice. Interactions between isogenic strains were independent of the amount of cells administered to mice. The cured derivative strain maintains its specific ability to adhere to epithelial cells of chicken crop.

Conjugal Transfer of Broad-Host-Range Plasmid pAMbeta1 into Enteric Species of Lactic Acid Bacteria.

The broad-host-range plasmid pAMbeta1, which codes for erythromycin and lincomycin resistance, was transferred by conjugation into Lactobacillus acidophilus, Lactobacillus reuteri, and Lactobacillus salivarius. A novel 17-megadalton plasmid molecule was detected in the transconjugants, confirming the introduction of pAMbeta1 into each species.

Tracheal intubation using alfentanil and no muscle relaxant: is the choice of hypnotic important?

Administration of alfentanil followed by propofol intravenously (IV) without neuromuscular blockade for induction of anesthesia provides adequate conditions for tracheal intubation. Other hypnotic drugs have not been thoroughly investigated in this regard. Accordingly, 140 ASA physical status I and II premedicated outpatients were randomly assigned to one of seven groups (n = 20/group). Patients in Groups I-VI received alfentanil 40 microg/kg followed by etomidate 0.3 mg/kg, propofol 2 mg/kg, or thiopental 4 mg/kg. One half of these patients (Groups II, IV, VI) also received lidocaine 1 mg/kg IV prior to the administration of the above drugs. Patients in group VII received d-tubocurarine 3 mg followed by thiopental 4 mg/kg and succinylcholine 1 mg/kg. Ninety seconds after induction, laryngoscopy and endotracheal intubation were attempted and graded. Patients in Group V (alfentanil/thiopental) were significantly (P < 0.05) more likely to have a clinically unacceptable response to intubation (55%) (e.g., vigorous coughing, purposeful movement, or requirement for succinylcholine to complete intubation) compared with patients who received propofol (35%) or etomidate (20%). Alfentanil/etomidate yielded intubation conditions comparable to those achieved with alfentanil/propofol and d-tubocurarine/thiopental/succinylcholine. Lidocaine appeared to improve intubating conditions, although this improvement did not reach statistical significance. The results suggest that healthy, premedicated patients with favorable airway anatomy who have received alfentanil 40 microg/kg can be reliably tracheally intubated 90 s after administration of propofol 2 mg/kg or etomidate 0.3 mg/kg.

Application of antimicrobial-producing lactic acid bacteria to control pathogens in ready-to-use vegetables.

Five psychrotrophic strains of lactic acid bacteria (Lactobacillus casei, Lact. plantarum and Pediococcus spp.) were isolated from 22 samples of commercial salads. These strains were shown to inhibit Aeromonas hydrophila, Listeria monocytogenes, Salmonella typhimurium and Staphylococcus aureus on MRS agar, in salads and in juice prepared from vegetable salads. Lactobacillus casei IMPCLC34 was most effective in reducing total mesophilic bacteria and the coliform group; Aer. hydrophila, Salm. typhimurium and Staph. aureus disappeared after 6 d of storage, while the counts for L. monocytogenes remained constant. The potential application of antimicrobial-producing lactic acid bacteria as biopreservatives of ready-to-use vegetables is suggested.

Resistencia única a Rifampicina en Tuberculosis asociada al Síndrome de Inmunodeficiencia Adquirida / Single resistance at Rifampicin in Tuberculosis associated at Acquired Immunodeficiency Syndrome

Se analizaron las historias clínicas de 12 pacientes de Tuberculosis con resistencia única a rifampicina asociada al Síndrome de Inmunodeficiencia Adquirida, cuyas muestras biológicas fueron estudiadas en el Laboratorio de Bacteriología "Dr. A. Cetrángolo" durante el período comprendido entre julio de 1991 y febrero de 1994. De los 12 pacientes, 11 eran hombres y 1 pertenecía al sexo femenino. La edad estuvo comprendida entre los 20 y 35 años. Los 10 pacientes con conducta de riesgo conocida eran adictos a drogas intravenosas. La tuberculosis fue pulmonar en 8 pacientes, extrapulmonar en 3 y localización combinada en 1 caso.