Mem Inst Oswaldo Cruz, 2004 Nov, 99(7), 749 - 52 Epub 2005 Jan 12. Drug susceptibility of Brazilian strains of Mycobacterium bovis using traditional and molecular techniques; Parreiras PM et al.; Transmission of Mycobacterium bovis from cattle to humans has been reported and can cause tuberculosis (Tb) and a problem in certain risk populations . Therefore, knowledge of resistance of M . bovis towards antibiotics used for therapy of human Tb could help avoiding cure delay and treatment cost increase when dealing with drug resistant organisms . We therefore evaluated the susceptibility of M . bovis isolates towards streptomycin, isoniazide, rifampicin, ethambutol, and ethionamide, the first line antibiotics for human Tb . Therefore, 185 clinical samples from cattle with clinical signs of tuberculosis were processed and submitted to culturing and bacterial isolates to identification and drug susceptibility testing using the proportion method . Among 89 mycobacterial strains, 65 were identified as M . bovis and none were resistant to any of the antibiotics used . Confirmation of present results by future studies, enrolling a large number of isolates and designed to properly represent Brazilian regions, may favor the idea of using isoniazide preventive therapy as part of a Tb control strategy in special situations . Also, nucleic acids from bacterial isolates were submitted to rifoligotyping, a recently described reverse hybridization assay for detection of mutations causing resistance towards rifampicin . Concordance between the conventional and the molecular test was 100%, demonstrating the use of such methodology for rapid evaluation of drug susceptibility in M . bovis. Aust Vet J, 2004 Dec, 82(12), 773 - 80 An evaluation of the role of antibodies to Actinobacillus pleuropneumoniae serovar 1 and 15 in the protection provided by sub-unit and live streptomycin-dependent pleuropneumonia vaccines; Tumamao JQ et al.; OBJECTIVE: To evaluate the serological response of pigs receiving either the Porcilis APP vaccine or a modified live vaccine based on a streptomycin-dependent (SD) strain of Actinobacillus pleuropneumoniae, and then challenged with an Australian isolate of A . pleuropneumoniae of either serovar 1 or 15 as a means of understanding the protection provided by both vaccines against serovar 1 but not against serovar 15 . DESIGN: The serological tests evaluated were serovar-specific polysaccharide ELISA tests (for serovar 1 and 15), ELISA tests for antibodies to three A . pleuropneumoniae toxins (ApxI, ApxII and ApxIII) as well as to a 42 kDa outer membrane protein (OMP), a haemolysin neutralisation (HN) assay and immunoblotting . The tests were used to detect antibodies in vaccinated pigs that had been shown to be protected against serovar 1 but not serovar 15 . RESULTS: In the polysaccharide antigen ELISA assays, both vaccines resulted in a significant rise in the titre in the serovar 1 ELISA but not the serovar 15 ELISA . The Porcilis APP vaccinated pigs showed a significant response in the ApxI, ApxIII and 42 kDa OMP ELISA . In the ApxII ELISA, all pigs tested (the Porcilis APP vaccinates and the controls) were positive on entry to the trial . In the HN assay, the Porcilis APP vaccinated pigs showed a significant response after one dose while the SD vaccinated pigs required two doses of vaccine before a marked rise in titre was induced . Immunoblotting revealed that neither vaccine generated antibodies that recognised the ApxIII produced by serovar 15 . CONCLUSIONS: The failure of these vaccines to provide protection against serovar 15 may be due to novel virulence factors possessed by serovar 15, significant differences between the ApxIII toxin of serovar 15 and those present in the Porcilis APP vaccine or failure by both vaccines to induce antibodies to the serovar 15 specific polysaccharide. Chir Organi Mov, 2004 Apr-Jun, 89(2), 177 - 80 Tuberculous trochanteric bursitis; Haritides J et al.; A case of tuberculous trochanteric bursitis in a 40-year old man is reported . The findings of x-rays, echo, bone scanning, CT scan and MRI are shown . After one month of anti-TB therapy the bursa was excised en bloc as well as the lateral part of the trochanter . Then a continuous suction irrigation system was applied for 3 weeks using streptomycin solution . The anti-TB therapy was continued for one year . The patient was asymptomatic with no signs of recurrence 5 years postoperatively. Yi Chuan, 2003 Jul, 25(4), 428 - 32 {The Occurrence of Actual F Factor Transfer during the Stationary-phase Mutation in Escherichia coli FC40.}; Zhang HB et al.; The experiment of adaptive mutation was performed by using Escherichia coli GM133 rif(r) as test cells and HB214 str(r) as scavenger cells.Transfer frequency between GM133 and HB214 was estimated,based on the number of revertants appeared on the selective plates when GM133 were killed by addition of M9 selective medium containing 100mug/mL of streptomycin at different time.After 30 minutes the cells of GM133 and HB214 were mixed,the estimated transfer frequency was about 0.07%,and two days,7.47%.After selection of 7 days,some HB214 cells with F' factor from GM133 cells and lac+ mutation were observed,but these cells failed to form the colonies which can be seen by the naked-eye.It was demonstrated that actual F' factor transfer events from test cells GM133 to scavenger cells HB214 occurred during the selection. Neurol Med Chir (Tokyo), 2004 Oct, 44(10), 562 - 7 Brucellar spine infection--four case reports; Turgut M et al.; Brucellosis can be difficult to diagnose because of the nonspecific and variable clinical picture . This systemic disease is still an important public health problem in the Mediterranean Basin . These four cases of brucellar spine infection originated from rural areas around Aydin, Turkey . The systemic complaints of fever, profuse sweating, malaise, polyarthromyalgia, and weight loss indicated the final diagnosis of brucellosis and presumptive diagnoses were made based on agglutination testing for brucellosis . Computed tomography and magnetic resonance imaging indicated signs of bone infection and soft tissue involvement . Good outcomes were obtained with prolonged treatment with antibrucellar drugs including streptomycin, rifampicin, and tetracycline . Early diagnosis is important and prompt antibrucellar chemotherapy is effective in most cases, but prolonged follow up is necessary in all patients with spinal brucellosis. Eur J Pediatr . 2004 Dec 23; {Epub ahead of print} Connatal tuberculosis in a very premature infant; Chang ML et al.; Connatal tuberculosis is increasing in incidence and the mortality and morbidity of this disease remains high . We report a 27-week-old, 896 g female premature infant who had mild respiratory distress syndrome after birth . She developed signs of infection, progressive pneumonia and atelectasis which did not respond to mechanical ventilation and antibiotics . At 41 days of age, Mycobacterium tuberculosis was isolated from the non-bronchoscopic bronchoalveolar lavage . The isolate was sensitive to isoniazid, rifampin, streptomycin, and pyrazinamide . Miliary tuberculosis was subsequently diagnosed in her mother on a chest X-ray film and sputum cultures . The infant was treated successfully with anti-tuberculosis drugs . She had normal growth and development at the chronological age of 20 months old . Conclusion:Connatal tuberculosis should be considered in premature infants with symptoms of sepsis refractory to antibiotics . Most premature infants with connatal tuberculosis have lung involvement, and non-bronchoscopic bronchoalveolar lavage can be a useful procedure to establish the diagnosis. Zhongguo Zhong Yao Za Zhi, 2003 Nov, 28(11), 1054 - 6 {Effect of tetradrine on electrophysilogic changes caused by rising of left ventricular preload in guinea pigs}; Wang XX et al.; OBJECTIVE: To investigate the changes of guinea pig heart electrophysiological properties caused by increasing left ventricular preload, and to assess the effects of tetradrine on these changes . METHOD: Working model preparation of guinea pig hearts in vitro was used, and the preload of left ventricle was increased by adjusting the prefusion pressure of left atria . The changes of heart electrophysiologic parameters including monophasic action potential duration (MAPD90), monophasic action potential amplitude (MAPA), effective refractory period (ERP) and ventricular fibrillation threshold (VFT) were observed before and after altering the preload of left ventricle, and compared in the absence and presence of tetradrine, streptomycin or verapamil . RESULT: The rising of left ventricular preload led to shortening of MAPD90, ERP, and to descent of MAPA, VFT (all P<0.01) . Both Tetradrine and streptomycin inhibited these changes of heart electrophysiologic parameters caused by elevation of left ventricular afterload (all P<0.01) . In contrast, verapamil had no effects on the preload-related electrophysiological changes (all P>0.05) . CONCLUSION: Electrophysiologic changes caused by increasing left ventricular preload may be inhibited by tetrandrine, through inhibition of stretch-activated ion channels. Ann N Y Acad Sci, 2004 Nov, 1026, 273 - 6 Local utilization of metacresolsulfonic acid combined with streptomycin in the treatment of actinomycosis; Silva LA et al.; The effectiveness of combining metacresolsufonic acid with streptomycin in the treatment of actinomycosis, diagnosed either clinically or in the laboratory, was evaluated in 12 bovines and 2 equines . Eighty-seven percent of treated animals were considered clinically cured and did not show any signs of relapse after a six-month follow-up period . Therapeutic diagnosis by clinical observation was the procedure of choice when it was not possible to obtain laboratory diagnosis. J Bacteriol, 2005 Jan, 187(1), 135 - 42 Dual transcriptional control of amfTSBA, which regulates the onset of cellular differentiation in Streptomyces griseus; Ueda K et al.; The amf gene cluster encodes a probable secretion system for a peptidic morphogen, AmfS, which induces aerial mycelium formation in Streptomyces griseus . Here we examined the transcriptional control mechanism for the promoter preceding amfT (PamfT) directing the transcription of the amfTSBA operon . High-resolution S1 analysis mapped a transcriptional start point at 31 nucleotides upstream of the translational start codon of amfT . Low-resolution analysis showed that PamfT is developmentally regulated in the wild type and completely abolished in an amfR mutant . The -35 region of PamfT contained the consensus sequence for the binding of BldD, a pleiotropic negative regulator for morphological and physiological development in Streptomyces coelicolor A3(2) . The cloned bldD locus of S . griseus showed high sequence similarity to the S . coelicolor counterpart . Transcription of bldD occurred constitutively in both the wild type and an A-factor-deficient mutant of S . griseus, which suggests that the regulatory role of BldD is independent of A-factor . The gel retardation assay revealed that purified BldD and AmfR recombinant proteins specifically bind PamfT . Overproduction of BldD in the wild-type cell conferred a bald phenotype (defective in aerial growth and streptomycin production) and caused marked repression of PamfT activity . An amfT-depleted mutant also showed a bald phenotype but PamfT activity was not affected . Both the bldD-overproducing wild-type strain and the amfT mutant were unable to induce aerial growth of an amfS mutant in a cross-feeding assay, which indicates that these strains are defective in the production of an active AmfS peptide . The results overall suggests that two independent regulators, AmfR and BldD, control PamfT activity via direct binding to determine the transcriptional level of the amf operon responsible for the production and secretion of AmfS peptide, which induces the erection of aerial hyphae in S . griseus. J Chromatogr A, 2004 Nov 26, 1058(1-2), 137 - 42 Determination of streptomycin and dihydrostreptomycin in milk and honey by liquid chromatography with tandem mass spectrometry; van Bruijnsvoort M et al.; Two liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were developed for the determination of streptomycin (STR) and its derivative dihydrostreptomycin (DHSTR) in milk and honey . These aminoglycoside antibiotics are used as veterinary drugs . In the EU, the presence of dihydro- and streptomycin residues in honey is forbidden, the maximum residue level (MRL) in milk is 200 microg/kg . The methods were optimised with regard to sensitivity and chromatographic efficiency, and validated by a procedure consistent with EU directive 2002/657 . Average recoveries and accompanying standard deviations were satisfactory . The limit of quantification of STR was 2 microg/kg in honey and 10 microg/kg in milk, of DHSTR it was a factor two lower . The precision of the milk analysis was improved by using STR as the internal standard for DHSTR and vice versa . In a survey of 186 honeys available on the Dutch market, 26% of the honeys of foreign origin were positive for (DH)STR . This occurence rate was consistent with previous surveys, but lower concentrations were found. Indian J Med Res, 2004 Nov, 120(5), 468 - 71 Drug susceptibility of Mycobacterium tuberculosis to primary antitubercular drugs by nitrate reductase assay; Sethi S et al.; BACKGROUND & OBJECTIVES: Rapid susceptibility testing of Mycobacterium tuberculosis strains is imperative for therapy selection but traditional drug susceptibility tests take weeks or are expensive . In this study we evaluated nitrate reductase assay which utilizes the detection of nitrate reduction as an indication of growth and therefore results can be obtained faster than by visual detection of colonies . METHODS: One hundred clinical isolates of M . tuberculosis were tested for four first line antitubercular drugs by nitrate reductase assay (NRA) and were compared with standard proportion method . The bacteria were inoculated on Lowenstein-Jensen (LJ) medium with primary antitubercular drugs and potassium nitrate was incorporated . After incubation for 7- 14 days, nitrate reduction indicating growth could be detected by colour change when reagents were added . RESULTS: Resistance of isolates as determined by both methods for isoniazid, rifampicin, streptomycin and ethambutol was 32, 35, 62 and 15 per cent respectively . Agreement between NRA and proportion method was 99 per cent for isoniazid and ethambutol . Complete agreement (100%) was found for rifampicin and streptomycin . Results were available in 7-14 days by NRA as compared to proportion method which takes 4-6 wk . INTERPRETATION & CONCLUSION: Nitrate reductase assay is a rapid and inexpensive method for susceptibility testing of M . tuberculosis for primary antitubercular drugs and could be an appropriate alternative to existing methods, particularly in resource-poor settings. J Med Microbiol, 2005 Jan, 54(1), 77 - 81 Safe susceptibility testing of Mycobacterium tuberculosis by flow cytometry with the fluorescent nucleic acid stain SYTO 16; Pina-Vaz C et al.; The time needed to obtain susceptibility results of Mycobacterium tuberculosis using classical methodologies is still too long, and flow cytometry is a promising technique in the setting of the clinical laboratory, giving fast results . A safe, reliable and rapid method to study susceptibility to streptomycin, isoniazide, rifampicin and ethambutol is described . Isolates of mycobacteria, grown for 72 h in the absence or presence of antimycobacterial drugs in the mycobacteria growth indicator tube (MGIT), were heat-killed, stained with SYTO 16 (a nucleic acid fluorescent stain that only penetrates cells with severe lesion of the membrane) and then analysed by flow cytometry . Sixteen strains with different susceptibility patterns were tested and an excellent correlation with the BACTEC MGIT 960 protocol for susceptibility was shown . In contrast to resistant strains, sensitive strains lose their cellular integrity after incubation with antimycobacterial drugs, allowing SYTO 16 to penetrate the cells . Comparing the intensity of fluorescence of Mycobacterium cells incubated with antimycobacterial drugs with that of drug-free cells, after staining with SYTO 16, it was possible to distinguish between sensitive, intermediate and resistant phenotypes . Other cytometric assays have been described for mycobacteria susceptibility testing but these have lower accuracy and safety . The described flow cytometric assay is a simple, fast, safe and accurate way to determine susceptibility of M . tuberculosis. Probl Tuberk Bolezn Legk, 2004, (10), 36 - 9 {Use of dissolved ozone in the treatment of experimental tuberculosis in mice}; Characterisation of rpsL et al.; Laboratory of Molecular Microbiology, Biomedical Research and Study Centre, University of Latvia, Ratsupites 1, Riga LV 1067, Latvia . tatjanap@biomed.lu.lv In order to characterise molecular mechanisms of first-line drug resistance in Mycobacterium tuberculosis and to evaluate the use of molecular markers of resistance (gene point mutations), we analysed 66 multi-drug-resistant (MDR) isolates from Latvian tuberculosis patients . They were all resistant to rifampin (RIF), isoniazid (INH) and streptomycin (SM), and 33 were resistant to ethambutol (EMB) . Enzymatic digestion by MboII and nucleotide sequencing of the rpsL gene fragment detected a single nucleotide substitution K43R in 40 (61%) of the 66 SM-resistant M . tuberculosis isolates . Of the other 26 SM-resistant isolates, 16 (24%) had mutations at positions 513A-->C and 516C-->T of the rrs gene and 10 (15%) had the wild-type sequence . The single-stranded DNA conformation polymorphism (SSCP) method was used to detect mutations in the embB gene associated with EMB resistance . Substitutions in the embB gene were found by SSCP analysis in 15 (45%) and by sequencing in 17 (52%) of the 33 EMB-resistant isolates . Surprisingly, SSCP revealed a nucleotide mutation at codon M306 in five (15%) of 33 in vitro EMB-susceptible MDR isolates. J Commun Dis, 2003 Jun, 35(2), 82 - 9 Prevalence of initial drug resistance to M . tuberculosis in new sputum positive RNTCP patients; Dhingra VK et al.; There is an increased prevalence of drug resistant M . tuberculosis strains and of these, multi drug resistant organisms are of particular concern . With the implementation of Revised National Tuberculosis Control Programme (RNTCP) allover the state of Delhi, Initial drug resistance (IDR) to Isoniazid and Rifampicin assumes great importance and needs to be monitored on a regular basis . We undertook to study the IDR against the first line essential drugs i.e . Isoniazid (H), Rifampicin (R), Ethambutol (E) and Streptomycin (S) from April 1999 to March 2000 in newly diagnosed sputum positive cases of pulmonary tuberculosis attending TB clinics under RNTCP in Delhi . A total of 157 consecutive new smear positive patients attending TB clinics under RNTCP were taken into the study . All sputum samples were subjected to culture and drug sensitivity tests on LJ medium after decontamination of samples by Petroff's method . Resistance was expressed as the percentage of colonies that grow on critical concentration of the drugs . To determine the proportion of resistance, the number of colonies on the control and the number of colonies on the drug medium were determined . A total of 94.77% samples were sensitive to the four first line essential drugs and IDR to any drug was 5.22% . The resistance to Rifampicin alone was nil but the resistance to Isoniazid alone was 2.24% . Combined resistance to both Rifampicin and Isoniazid was 2.98 % . The incidence of resistance to first line drugs in tuberculosis is not very high among new sputum positive patients attending TB clinics under RNTCP. Indian J Chest Dis Allied Sci, 2004 Jul-Sep, 46(3), 171 - 7 Antituberculosis drug resistance and associated risk factors in the European section of Turkey; Karabay O et al.; BACKGROUND: We carried out this study in order to establish the prevalence of antituberculosis drug resistance in Mycobacterium tuberculosis strains and to determine risk factors for the development of resistance in Trakya region of Turkey . METHODS: Pattern of drug resistance in 214 M . tuberculosis isolates from patients with tuberculosis treated at the regional tuberculosis dispensaries were included in the study . RESULTS: Isolates of 105 (49.1%) were resistant to only one drug, and 62 (29.0%) were resistant to more than one drug . The total resistance rates to streptomycin, isoniazid, rifampicin, ethambutol and isoniazid + rifampicin were 29.0%, 27.1%, 21.5%, 10.3% and 11.6%, respectively . The secondary resistance rates in all drugs and combinations were higher than primary resistance rates (p<0.001) . Step wise logistic regression revealed that (i) non-compliance with treatment increases the chances of development of resistance by 15 times {p<0.00001, 95% confidence intervals (95% CI) : 4.16 to 56.70}, and (ii) a regimen of inadequate treatment increases the chance of development of drug resistance by 10.5 times (p<0.01, 95% CI=2.66 to 49.80) . CONCLUSIONS: We propose that specially trained physicians should institute antituberculosis therapy and medication should be practiced under direct observation in this region. J Neurobiol . 2004 Nov 16; {Epub ahead of print} Aminoglycosides block the Kv3.1 potassium channel and reduce the ability of inferior colliculus neurons to fire at high frequencies; Liu SQ et al.; The Kv3.1 potassium channel is expressed at high levels in auditory nuclei and contributes to the ability of auditory neurons to fire at high frequencies . We have tested the effects of streptomycin, an agent that produces progressive hearing loss, on the firing properties of inferior colliculus neurons and on Kv3.1 currents in transfected cells . We found that in inferior colliculus neurons, intracellular streptomycin decreased the current density of a high threshold, noninactivating outward current and reduced the rate of repolarization of action potentials and the ability of these neurons to fire at high frequencies . Furthermore, potassium current in CHO cells transfected with the Kv3.1 gene was reduced by 50% when cells were cultured in the presence of streptomycin or when streptomycin was introduced intracellularly in the pipette solution . In the presence of intracellular streptomycin, the activation rate of Kv3.1 current increased and inhibition by extracellular TEA become voltage-dependent . The data indicate that streptomycin inhibits Kv3.1 currents by inducing a conformational change in the Kv3.1 channel . The hearing loss caused by aminoglycoside antibiotics may be partially mediated by their inhibition of Kv3.1 current in auditory neurons . (c) 2004 Wiley Periodicals, Inc . J Neurobiol, 2005. Ann Pharmacother, 2005 Jan, 39(1), 165 - 8 Epub 2004 Nov 16. A rare case of streptomycin-induced toxic epidermal necrolysis in a patient with tuberculosis: a therapeutic dilemma; Hmouda H et al.; OBJECTIVE: To report a case of streptomycin-induced toxic epidermal necrolysis (TEN) . CASE SUMMARY: A 55-year-old woman was admitted for treatment of active pulmonary tuberculosis (TB) . She was given standard oral anti-TB chemotherapy including isoniazid, rifampin, pyrazinamide, and streptomycin . On the fourth day of therapy, she experienced high fever at 39 degrees C, chills, vomiting, pruritus, and diffuse erythema, followed by extensive bullae formation and skin denudation . Diagnosis of TEN was considered, and all anti-TB drugs were discontinued . Skin biopsy disclosed complete epidermal necrosis with dermal-epidermal cleavage and absence of inflammatory infiltrate, highly suggestive of TEN . The patient was transferred to the intensive care unit . Her general condition and skin lesions improved . A staged-fashion exposure test to the 4 anti-TB drugs allowed the incrimination of streptomycin as the offending agent . DISCUSSION: Anti-TB drugs, mainly rifampin, ethambutol, and isoniazid, have been incriminated in TEN . Streptomycin-induced TEN remains an extremely rare event . However, minor allergic skin reactions (rash, urticaria) have been described with this drug . Our patient presents a rare case of streptomycin-related TEN . Even though dangerous, a step-wise exposure test was necessary to allow safe treatment of active pulmonary TB . It also provided a strong argument of a cause-effect relationship between TEN and streptomycin . An objective causality assessment using the Naranjo rating scale revealed that the adverse drug event was highly probable . CONCLUSIONS: Streptomycin should be added to the list of drugs that induce TEN. Cell Calcium, 2005 Jan, 37(1), 69 - 80 Inhibition of the Na+-H+ exchanger with cariporide abolishes stretch-induced calcium but not sodium accumulation in mouse ventricular myocytes; Kondratev D et al.; We address the question whether activation of the sodium-proton exchanger (NHE) does contribute to the stretch-induced accumulation of intracellular sodium and calcium in mouse ventricular myocytes . NHE-blocker cariporide (10 microM) were applied to the bath for 10 min . Axial stretch was applied for 2 min by increasing the distance between an adherent glass stylus and the patch pipette by 20% . Myocytes (stimulated at 3 Hz) were shock-frozen in diastole and the membrane currents monitored till cryofixation . Controls were treated identically, but not stretched . Total sodium and calcium concentrations ({Na}, {Ca}=sum of free and bound Na and Ca) were measured by electron probe microanalysis (EPMA) in peripheral and central cytosol, mitochondria, nucleus and nuclear envelope . Cariporide did not reduce the stretch-activated negative current . The stretch-induced rise in {Na} was not different in the presence and in the absence of cariporide . Cariporide significantly reduced diastolic {Ca} in the cytosol of stretched myocytes . Since cariporide does not prevent the stretch-induced {Na} accumulation, we suggest that not NHE but the stretch-activated streptomycin-sensitive current I(SAC) causes the well documented stretch-induced {Na} accumulation . The discovery that cariporide prevents the stretch-induced rise in cytosolic {Ca} demonstrates an important additional effect of the drug on calcium handling. J Physiol . 2004 Nov 4; {Epub ahead of print} Effects of stretch-activated channel blockers on {Ca2+}i and muscle damage in the mdx mouse; Yeung EW et al.; The mdx mouse lacks dystrophin and is a model of human Duchenne muscular dystrophy . Single mdx muscle fibres were isolated and subjected to a series of stretched (eccentric) contractions while measuring intracellular calcium ({Ca2+}i) with fluo-3 and confocal microscopy . Following the stretched contractions there was a slow rise in resting {Ca2+}i and after 30 min both the {Ca2+} i during a tetanus (tetanic {Ca2+}i ) and the tetanic force were reduced . Two blockers of stretch-activated channels, streptomycin and the spider venom toxin GsMTx4, prevented the rise of resting {Ca2+}i and partially prevented the decline of tetanic {Ca2+}i and force . Reducing extracellular calcium to zero also prevented the rise in resting {Ca2+}i and prevented some of the decline in tetanic {Ca2+}i and force . Patch clamping experiments identified a stretch-activated channel in both wild-type and mdx muscle membranes which was blocked by GsTMx4 . These data suggest that blockers of stretch-activated channels can reduce some components of muscle damage after stretched contractions in isolated mdx muscle . We therefore tested whether streptomycin, added to the drinking water, was capable of reducing muscle damage in intact mdx mice . Mdx mice show a period of muscle damage from 20-40 days of life of which the appearance of centralized nuclei is one marker . We measured the frequency of central nuclei in control mdx mice compared to streptomycin-treated mdx mice and showed that the incidence of central nuclei was significantly reduced by streptomycin treatment . This data suggests that blockers of stretch-activated channels may protect against muscle damage in the intact mdx mouse. J Obstet Gynaecol, 1997, 17(2), 119 - 22 Gynaecological tuberculosis since 1951; Sutherland AM; A study has been made of 711 patients with proved gynaecological tuberculosis investigated between 1 January 1951 and 31 December 1994 . The main presenting symptoms in order of frequency were infertility, pelvic pain, excessive menstrual loss and amenorrhoea . The average age at first attendance was 31 years, only 16% of married women had been pregnant and palpable adnexal masses were found in 47% . Eight drug programmes were used, the best results being obtained with streptomycin, para-aminosalicyclic (PAS) acid and isoniazid or with rifampicin, ethambutol and isoniazid . Toxic drug reactions occurred in 114 patients, the drugs responsible in the majority being streptomycin and PAS . Where drug treatment failed, surgery under further drug cover was employed in 85 patients, with no deaths and no fistulae . After treatment, there were 95 pregnancies in 57 patients . In recent years there has been a rise in the incidence of tuberculosis in general in the Western world . Many factors play a part in this, by far the most important being the spread of AIDS. J Assoc Res Otolaryngol, 2004 Sep, 5(3), 323 - 36 Epub 2004 Aug 12. Posture, head stability, and orientation recovery during vestibular regeneration in pigeons; Dickman JD et al.; Compensatory behavior such as oculomotor, gaze, and postural responses that occur during movement largely depend upon a functioning vestibular system . In the present study, the initial loss and subsequent recovery of postural and head stability in pigeons undergoing vestibular regeneration were examined . Adult pigeons were trained to manipulate a straight run chamber to peck an illuminated key for fluid reward . Six behavioral measures assessing performance, posture, and head stability were quantified . These included run latency, steps (walking), path negotiation (lane changes), gaze saccades, head bobs, and head shakes . Once normative values were obtained for four birds, complete lesion of all receptor cells and denervation of the epithelia in the vestibular endorgans were produced using a single intralabyrinthine application of streptomycin sulfate . Each bird was then tested at specific times during regeneration and the same behavioral measures examined . At 7 days post-streptomycin treatment (PST), all birds exhibited severe postural and head instability, with tremors, head shakes, staggering, and circling predominating . No normal trial runs, walking, gaze saccades, or head bobs were present . Many of these dysfunctions persisted through 3-4 weeks PST . Gradually, tremor and head shakes diminished and were replaced with an increasing number of normal head bobs during steps and gaze saccades . Beginning at 4 weeks PST, but largely inaccurate, was the observed initiation of directed steps, less staggering, and some successful path negotiation . As regeneration progressed, spatial orientation and navigation ability increased and, by 49 days PST, most trials were successful . By 70 days PST, all birds had recovered to pretreatment levels . Thus, it was observed that ataxia must subside, coincident with normalized head and postural stability prior to the recovery of spatial orientation and path navigation recovery . Parallels in recovery were drawn to hair cell regeneration and afferent responsiveness, as inferred from present results and those in other investigations. J Mol Biol, 2004 Nov 5, 343(5), 1183 - 94 Conformational changes of the small ribosomal subunit during elongation factor G-dependent tRNA-mRNA translocation; Peske F et al.; Translocation, a coordinated movement of two tRNAs together with mRNA on the ribosome, is catalyzed by elongation factor G (EF-G) . The reaction is accompanied by conformational rearrangements of the ribosome that are, as yet, not well characterized . Here, we analyze those rearrangements by restricting the conformational flexibility of the ribosome by antibiotics binding to specific sites of the ribosome . Paromomycin (Par), viomycin (Vio), spectinomycin (Spc), and hygromycin B (HygB) inhibited the tRNA-mRNA movement, while the other partial reactions of translocation, including the unlocking rearrangement of the ribosome that precedes tRNA-mRNA movement, were not affected . The functional cycle of EF-G, i.e . binding of EF-G.GTP to the ribosome, GTP hydrolysis, Pi release, and dissociation of EF-G.GDP from the ribosome, was not affected either, indicating that EF-G turnover is not coupled directly to tRNA-mRNA movement . The inhibition of translocation by Par and Vio is attributed to the stabilization of tRNA binding in the A site, whereas Spc and HygB had a direct inhibitory effect on tRNA-mRNA movement . Streptomycin (Str) had essentially no effect on translocation, although it caused a large increase in tRNA affinity to the A site . These results suggest that conformational changes in the vicinity of the decoding region at the binding sites of Spc and HygB are important for tRNA-mRNA movement, whereas Str seems to stabilize a conformation of the ribosome that is prone to rapid translocation, thereby compensating the effect on tRNA affinity. Med Arh, 2004, 58(3), 183 - 4 {Unusual therapeutic approach in treatment of pulmonary tuberculosis in six year old girl}; Saracevic E et al.; We report about interesting case of treatment of pulmonary tuberculosis in 6-year old girl . Antituberculotic therapy induced toxic hepatitis (rise of transaminases, indirect hiperbilirubinemia) . She was treated with streptomycin, etambutol, ciprofloxacyn and systemic corticosteroides . Two gallbladder stones and high-density sediment were diagnosed after ultrasound examination which could be a side effect of the therapy or previous findings. Probl Tuberk Bolezn Legk, 2004, (8), 35 - 41 {Drug resistance of Mycobacterium tuberculosis of the genotype Beijing in imprisonment places in the Arkhangelsk Region}; Tungusova OS et al.; The paper describes the molecular genetic characteristics of M . tuberculosis (MT) strains isolated from 114 patients with pulmonary tuberculosis in the penitentiary system of the Arkhangelsk Region . The sensibility to the first-line antituberculous drugs was determined by the radiometric technique BACTEC; rpoB gene mutations were identified by Inno-Lipa, genotyping was made by the RFLPA assay and spoligotyping . The spread of MT of the genotype Beijing in the penitentiary system was found to be 76.3% . The high rate (79.8%) of the clustering pattern is indicative of active transmission of tuberculosis . The transmission of the Beijing genotype MBT strains was higher (96.6%) than that of other genetypes (25.9%) . A multifactorial analysis has shown that streptomycin resistance is independently associated with the infection with the strain of MBT of the genotype Beijing (p = 0.02); the circulation and active transmission of Beijing among the population present a human health threat and an endemic risk from the spread of drug-resistant pulmonary tuberculosis. Laryngoscope, 2004 Sep, 114(9), 1630 - 2 Effect of edaravone on streptomycin-induced vestibulotoxicity in the Guinea pig; Horiike O et al.; OBJECTIVES/HYPOTHESIS: The effect of topical administration of edaravone to the inner ear was investigated in guinea pigs with streptomycin-induced vestibulotoxicity . METHODS: Vestibulotoxicity was induced in 20 animals by delivery of streptomycin into the inner ear through osmotic pump for 24 hours . Edaravone (n = 8, systemic administration group) or saline (n = 6, control group) was injected intraperitoneally once a day for 7 days or edaravone-soaked Gelfoam was placed on the round window before wound closure (n = 6, topical administration group) . RESULTS: Yaw head tilt and spontaneous nystagmus were observed in all animals after the operation . The number of spontaneous nystagmus beats in the topical administration group was statistically less than that in other two groups at 12, 18, and 24 hours after the operation . CONCLUSION: The study results suggest that topical administration of edaravone better suppresses streptomycin-induced vestibulotoxicity than systemic administration. Monaldi Arch Chest Dis, 2004 Jan-Mar, 61(1), 65 - 70 Drug-resistance of Mycobacterium tuberculosis in Patras, Greece; Trakada G et al.; BACKGROUND: The global distribution of drug resistant tuberculosis reflects the quality of tuberculosis control worldwide and is still a major public health-problem . To our knowledge, no data exists in literature about resistance to anti-tuberculosis drugs in Greece . METHODS: The aim of this study was to determine the prevalence of resistance to the main anti-tuberculosis drugs in newly and previously treated tuberculosis patients, in the region of Patras, in Greece and to evaluate the contribution of foreign-born and human immunodeficiency virus (HIV) positive cases to drug resistance . RESULTS: A total of 207 initial isolates of Mycobacterium tuberculosis were analysed . All clinical specimens submitted for cultural diagnosis were collected before chemotherapy commenced . The age of the patients varied from 16 to 78 years old . Sex distribution was 144 males (69.56%) and 63 females (30.43%) . Nineteen patients (9.17%) were recent immigrants in Greece . All the patients were HIV-negative . One hundred-one initial isolates of Mycobacterium tuberculosis (48.79%) were susceptible to all first-line anti-tuberculosis drugs, isoniazid, streptomycin, ethambutol and rifampicin . The prevalence of primary mono- and poly-drug resistance was lower (isoniazid 5.79%, streptomycin 4.34%, ethambutol 1.93%, isoniazid + streptomycin 3.38%, isoniazid + ethambutol 0.9%, isoniazid + streptomycin + ethambutol 1.44%) when compared with the prevalence of secondary (acquired) mono- and poly-drug resistance (isoniazid 8.69%, streptomycin 10.14%, ethambutol 3.86%, isoniazid + streptomycin 3.86%, isoniazid + rifampicin 1.44%, isoniazid + streptomycin + ethambutol 4.83%, isoniazid + streptomycin + rifampicin 1.44%) . No primary mono - resistance to rifampicin, or primary multi - drug resistance (defined here as Mycobacterium tuberculosis resistant to at least isoniazid and rifampicin) were observed . CONCLUSIONS: According to our data, overall drug resistance was high . Fortunately, multi-drug resistance tuberculosis was not common but special efforts are needed to monitor the prevalence of drug resistance and to ensure adequate treatment in all the population. Clin Infect Dis, 2004 Sep 1, 39(5), e35 - 45 Epub 2004 Aug 11. New times for an old disease: intracranial mass lesions caused by Mycobacterium tuberculosis in 5 HIV-negative African immigrants; Ripamonti D et al.; BACKGROUND: The tuberculosis epidemic is still a global emergency, and its spread in the past 20 years has been fueled by the acquired immune deficiency syndrome pandemic and increasing drug resistance . International travel and migration may increase the incidence of tuberculosis in industrialized countries . METHODS: We reviewed the clinical charts of patients admitted to the infectious diseases unit of Ospedali Riuniti (Bergamo, Italy) to identify patients with intracranial mass lesions caused by Mycobacterium tuberculosis . RESULTS: During the past 6.5 years, 5 of 30 patients with a mass of infectious origin in the brain had tuberculous brain lesions diagnosed . All 5 were human immunodeficiency virus (HIV)-negative adults and African immigrants . No patient had concomitant meningitis, 1 had a concomitant pulmonary disease, and 3 subjects reported a past history of tuberculosis . At presentation, no patient had fever and 3 had seizures . Examination of cerebrospinal fluid revealed normal findings for 4 of 4 subjects, and neuroimaging showed multiple intracranial mass lesions in 4 of 5 patients . The diagnosis was definite for 2 subjects (based on analysis of brain specimens) and presumptive for 3 subjects (1 had concomitant pulmonary tuberculosis, and 2 had clinical response to therapy) . Results of susceptibility tests for M . tuberculosis were available for 2 patients: both isolates were resistant to isoniazid, and 1 was also resistant to streptomycin . Duration of medical treatment ranged from 11 to 23 months, and 2 subjects underwent surgical procedures at the time of diagnosis . All 5 patients recovered . CONCLUSIONS: Clinicians in western countries should consider the possible role of tuberculosis in causing mass lesions in the brain, particularly in immigrants from regions where tuberculosis is endemic. J Antimicrob Chemother, 2004 Oct, 54(4), 722 - 9 Epub 2004 Sep 08. Effect of n-octanesulphonylacetamide (OSA) on ATP and protein expression in Mycobacterium bovis BCG; Parrish NM et al.; OBJECTIVE: To determine the effect on BCG of n-octanesulphonylacetamide (OSA), a novel compound of the class beta-sulphonylcarboxamides, which has potent in vitro activity against pathogenic mycobacteria . METHODS AND RESULTS: The effect of OSA in BCG was examined using two-dimensional protein electrophoresis . Treatment of BCG with OSA resulted in overexpression of two proteins identified as the b-subunit of ATP synthase (Rv1306) and a 17 kDa heat shock protein (Rv0251c) . {35S}Methionine pulse-labelling revealed that overexpression occurred within as little as 3.5 h post-exposure . These results were confirmed by RT-PCR . ATP levels decreased in OSA-treated BCG at 5 min, and 1, 3 and 24 h, with a 64%, 45%, 54% and 73% reduction in ATP, respectively . Only dicyclohexylcarbodiimide (DCCD), a known ATP synthase inhibitor, had a similar effect . No appreciable difference in ATP level was observed in BCG treated with standard antimycobacterial drugs, additional respiratory chain inhibitors or a fatty acid synthase inhibitor at a comparable time-point . Protein synthesis decreased within 5 min of exposure to OSA (56%), DCCD (74%) and thenoyltrifluoroacetone (TTFA) (77%) . Ethanol (2.3%) potentiated the activity of OSA . In contrast, no synergic effect was observed with streptomycin and ethanol . Mycolic acid levels decreased 79% with DCCD, 46% with TTFA, a complex II inhibitor, and 43% with OSA compared with untreated controls . CONCLUSIONS: Our results suggest that OSA may interfere directly or indirectly with ATP synthase and possibly other components of the mycobacterial respiratory chain . These effects may hinder energy production, leading to interruption in the synthesis of large macromolecules including proteins and mycolic acids. Zh Mikrobiol Epidemiol Immunobiol, 2004 May-Jun, (3), 95 - 8 {Decreased resistance of multiresistant mycobacteria to isoniazid during the treatment of experimental tuberculosis with ozone and isoniazid}; Belianin II; Mycobacteria (MB) of the clinical strain resistant to streptomycin, isoniazid (IN), rifampicin and kanamycin were injected intravenously into 68 BALB/c mice . The animals were divided into 5 groups: two control groups 0 and 1 (intact and infected without subsequent treatment), group 2 (treated with IN), group 3 (treated with IN and injected intraperitoneally with dissolved ozone, or dO3), group 4 (injected with dO3) . The animals started to die by month 4 after the infection . By month 5 all mice died with the exception of intact mice and those treated with dO3) . By month 4 the study of MB cultures isolated from the lungs revealed a decrease in their resistance to IN in the groups undergoing treatment with dO3 . Hepatic and splenic lesions were observed after treatment with IN only were greater than in the absence of treatment . After the use of IN + dO3 such lesions were the least . The mechanism of a decrease in the medicinal resistance of MB under the action of dO3 and the expediency of the simultaneous use IN and dO3 in cases of the unknown medicinal resistance of MB are discussed. J Chemother, 2004 Aug, 16(4), 347 - 9 The contribution of a novel ribosomal S12 mutation to aminoglycoside resistance of Escherichia coli mutants; Gill AE et al.; This study characterised the contribution of a novel ribosomal S12 mutation to aminoglycoside resistance in Escherichia coli via step-wise mutation analysis . Mutants of E . coli NCTC 10418 were selected in four separate progressive series (I-IV) on plates containing increasing aminoglycoside (streptomycin, neomycin, gentamicin, tobramycin, and kanamycin) concentrations . Minimum inhibitory concentrations (MICs) of these aminoglycosides were established for the most resistant mutants in each series . There was no cross-resistance between streptomycin and the other aminoglycosides tested; however there was cross-resistance between the neomycin, tobramycin and kanamycin resistant mutants . DNA sequencing of a 423bp region of the rpsL gene encoding S12 revealed a novel Lys87-->Glu mutation in the streptomycin selected resistant mutants, while there were no S12 mutations in resistant mutants resulting from selection with neomycin, gentamicin, and tobramycin and kanamycin. Int J Antimicrob Agents, 2004 Sep, 24(3), 304 - 6 Drug susceptibility testing of Mycobacterium tuberculosis with nitrate reductase assay; Coban AY et al.; The nitrate reductase assay (NRA) was evaluated for susceptibility testing of Mycobacterium tuberculosis using 80 clinical isolates of M . tuberculosis and H37Rv as a control strain . All isolates were tested by the proportion method and the NRA for isoniazid (INH), rifampicin (RIF), streptomycin (STR) and ethambutol (ETM) . The proportion method was carried out according to NCCLS on Lowenstein-Jensen (LJ) medium and the NRA on LJ medium containing 1000 microg/ml potassium nitrate (KNO(3)) . After incubation for 7, 10, 14 and 21 days, Griess reagent was added to each LJ medium and nitrate reduction was determined by a colour change . Comparing the NRA with the proportion method, sensitivities were 100, 100, 82.1 and 92.2% for INH, RIF, STR and ETM, respectively . Specificities were 100, 100, 92.3 and 100% for INH, RIF, STR and ETM, respectively . The results of 2, 22 and 56 isolates were obtained after 7, 10 and 14 days, respectively . The proportion method result were read at 21-28 days . The NRA is rapid, inexpensive and easy to perform . Our results indicated that the NRA is suitable for the early determination of INH and RIF resistance in countries where sophisticated procedures are not always available. J Antibiot (Tokyo), 2004 Jun, 57(6), 383 - 9 Generation of Streptomyces globisporus SMY622 strain with increased landomycin E production and it's initial characterization; Gromyko O et al.; Landomycin E (LaE) overproducing strain Streptomyces globisporus SMY6222 has been developed using UV induced mutagenesis and selection for streptomycin resistance . SMY622 has been shown by HPLC to produce 200-fold higher amounts of LaE when comparing with parental strain . The levels of transcription of regulatory gene lndI and oxygenase gene lndE are two times higher in the mutant than in the wild type . Gene rpsL for ribosomal protein S12 from SMY622 was shown to contain point mutation K43R . Possible reasons for increased LaE synthesis in SMY622 are discussed. Biol Chem, 2004 Jul, 385(7), 615 - 21 GAPDH enhances group II intron splicing in vitro; Bock-Taferner P et al.; Group II introns are autocatalytic RNAs which self-splice in vitro . However, in vivo additional protein factors might be involved in the splicing process . We used an affinity chromatography method called 'StreptoTag' to identify group II intron binding proteins from Saccharomyces cerevisiae . This method uses a hybrid RNA consisting of a streptomycin-binding affinity tag and the RNA of interest, which is bound to a streptomycin column and incubated with yeast protein extract . After several washing steps the bound RNPs are eluted by addition of streptomycin . The eluted RNPs are separated and the proteins identified by mass-spectrometric analysis . Using crude extract from yeast in combination with a substructure of the bl1 group II intron (domains IV-VI) we were able to identify four glycolytic enzymes; glucose-6-phosphate isomerase (GPI), 3-phosphoglycerate kinase (PGK), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and triosephosphate isomerase (TPI) . From these proteins GAPDH increases in vitro splicing of the bl1 group II intron by up to three times . However, in vivo GAPDH is not a group II intron-splicing factor, since it is not localised in yeast mitochondria . Therefore, the observed activity reflects an unexpected property of GAPDH . Band shift experiments and UV cross linking demonstrated the interaction of GAPDH with the group II intron RNA . This novel activity expands the reaction repertoire of GAPDH to a new RNA species. J Heart Lung Transplant, 2004 Aug, 23(8), 1003 - 7 Antagonists of stretch-activated ion channels restore contractile function in hamster dilated cardiomyopathy; Nicolosi AC et al.; BACKGROUND: Stretch-activated ion channels (SACs) mediate abnormal ion currents in dilated cardiomyopathy (DCM), but their role in the contractile defect of DCM is undefined . We hypothesized that SAC antagonists would enhance contractile function in a hamster model of DCM . METHODS: Left ventricular papillary muscles from Syrian hamsters with a genetic DCM (n = 26), and from non-myopathic controls (n = 26), were superfused and stimulated to contract . Maximum active force (F(max); milli-Newtons per square millimeter) was determined before (baseline) and after subjecting the muscle to a 60-minute period of overstretch (resting length associated with a 20% decay in baseline maximum force {F(max)}) . Gadolinium (10 micromol/liter) and streptomycin (40 micromol/liter) were used separately to antagonize SACs . RESULTS: In the absence of SAC antagonist, baseline F(max) was greater in controls (1.79 +/- 0.26) vs DCM (0.69 +/- 0.12; p < 0.05) . Overstretch caused further decrease in F(max) in DCM (to 0.50 +/- 0.08; p = 0.03 vs baseline), but not in controls . The SAC antagonists increased baseline F(max) in DCM to equal that of untreated controls (gadolinium 1.64 +/- 0.34, streptomycin 2.13 +/- 0.33), but neither agent increased baseline F(max) in controls (gadolinium 1.91 +/- 0.20, streptomycin 2.25 +/- 0.49) . Both agents abolished the stretch-induced decrease in contractile function in DCM . CONCLUSIONS: Antagonists of SACs enhance contractile function in DCM to equal that of normal controls, and abolish sensitivity to further stretch . They do not alter contractile function in normal muscle . These data suggest an important role of SACs in the contractile dysfunction of DCM and further suggest that SAC antagonists may represent novel therapy in heart failure. Int J Tuberc Lung Dis, 2004 Aug, 8(8), 1032 - 5 Molecular characterisation of streptomycin-resistant Mycobacterium tuberculosis strains isolated in Poland; Brzostek A et al.; Primary drug resistance of Mycobacterium tuberculosis strains in Poland increased two-fold between 1997 and 2000 . Among 3705 drug-resistant strains investigated in 2000, 169 were resistant to streptomycin alone or in combination with isoniazid, rifampicin and/or ethambutol . The molecular basis of streptomycin resistance for 88 (52%) of these strains in comparison with 15 susceptible controls was determined . The most prevalent mutation was the single substitution Lys43Arg in the rpsL gene, found in 30.7% of the strains analysed . However, as many as 51% of the strains investigated carried no mutation in the rpsL or rrs genes . The multiple mutations present in two Beijing family strains were also identified. J Comp Physiol A Neuroethol Sens Neural Behav Physiol, 2004 Sep, 190(9), 747 - 58 Epub 2004 Aug 05. Lateral line-mediated rheotactic behavior in tadpoles of the African clawed frog (Xenopus laevis); Simmons AM et al.; Tadpoles (Xenopus laevis) have a lateral line system whose anatomical structure has been described, but whose functional significance has not been closely examined . These experiments tested the hypothesis that the lateral line system is involved in rheotaxis . Tadpoles in developmental stages 47-56 oriented toward the source of a water current . Orientation was less precise after treatment with cobalt chloride or streptomycin, but was similar to that of untreated animals after exposure to gentamicin . In no current conditions, tadpoles exhibited a characteristic head-down posture by which they held themselves in the water column at an angle around 45 degrees . This body posture became significantly less tilted in the presence of water current . Treatment with cobalt chloride or streptomycin increased the angle of tilt close to that seen in no current conditions, while gentamicin treatment tended to decrease tilt angle . The data are consistent with anatomical and physiological findings that tadpole neuromasts are similar to superficial, but not canal, neuromasts in fishes, and they suggest that the lateral line system is involved in both directional current detection and current-related postural adjustments in Xenopus. Clin Exp Pharmacol Physiol, 2004 Aug, 31(8), 551 - 6 Stretch-activated channels in stretch-induced muscle damage: role in muscular dystrophy; Yeung EW et al.; 1 . Stretch-induced muscle injury results in the damage that causes reduced force and increased membrane permeability . This muscle damage is caused, in part, by ionic entry through stretch-activated channels and blocking these channels with Gd3+ or streptomycin reduces the force deficit associated with damage . 2 . Dystrophin-deficient muscles are more susceptible to stretch-induced muscle injury and the recovery from injury can be incomplete . We have found that Na+ entry associated with stretch-induced injury is enhanced in dystrophin-deficient muscles and that blockers of stretch-activated channels are capable of preventing ionic entry and reducing muscle damage . 3 . A model is presented that proposes links between stretch-induced injury, opening of stretch-activated channels, increased levels of intracellular ions and various forms of muscle damage . Although changes in Na+ accompany stretch-induced muscle injury, we believe that changes in Ca2+ probably have a more central role in the damage process. Clin Exp Pharmacol Physiol, 2004 Aug, 31(8), 485 - 93 Skeletal muscle function: role of ionic changes in fatigue, damage and disease; Allen DG; 1 . Repeated activity of skeletal muscle causes a variety of changes in its properties: muscles become weaker with intense use (fatigue), may feel sore and weak after repeated contractions involving stretch and can degenerate in some disease conditions . The present review considers the role of early ionic changes in the development of each of these conditions . 2 . Single fibre preparations of mouse muscle were used to measure ionic changes following activity induced changes in function . Single fibres were dissected with intact tendons and stimulated to produce force . Fluorescent indicators were microinjected into the fibres to allow simultaneous ionic measurements with determination of mechanical performance . 3 . One theory to explain muscle fatigue is that fatigue is caused by the accumulation of lactic acid, producing an intracellular acidosis that inhibits the myofibrillar proteins . In contrast, we found that during repeated tetani there was little or no pH change, but that failure of calcium release was a major contributor to fatigue . Currently, it is proposed that precipitation of calcium and phosphate in the sarcoplasmic reticulum contributes to the failure of calcium release . 4 . Muscles can be used to shorten and produce force or they can be used to de-accelerate loads (stretched or eccentric contractions) . One day after intense exercise involving stretched contractions, muscles are weak, sore and tender, and this damage can take a week to recover . In this condition, sarcomeres are disorganized and there are increases in resting intracellular Ca2+ and Na+ . Recently, we demonstrated that the elevation of Na+ occurs through a stretch-activated channel that can be blocked by either gadolinium or streptomycin . Preventing the increase in {Na+}i with gadolinium also prevented part of the muscle weakness after stretched contractions . 5 . Duchenne muscular dystrophy is a lethal degenerative disease of muscles in which the protein dystrophin is absent . Dystrophic muscles are more susceptible to stretch-induced muscle damage and the stretch-activated channel seems to be one pathway for the increases in intracellular Ca2+ and Na+ that are a feature of this disease . We have shown recently that blockers of the stretch-activated channel can minimize some of the short-term damage in muscles from the mdx mouse, which also lacks dystrophin . Currently, we are testing whether blockers of the stretch-activated channels given systemically to mdx mice can protect against some features of the disease. Nucleic Acids Res, 2004 Aug 04, 32(14), 4119 - 26 Print 2004. A minimum structure of aminoglycosides that causes an initiation shift of trans-translation; Konno T et al.; Trans-translation is an unusual translation in which transfer-messenger RNA plays a dual function--as a tRNA and an mRNA--to relieve the stalled translation on the ribosome . It has been shown that paromomycin, a typical member of a 4,5-disubstituted class of aminoglycosides, causes a shift of the translation-resuming point on the tmRNA by -1 during trans-translation . To address the molecular basis of this novel effect, we examined the effects of various aminoglycosides that can bind around the A site of the small subunit of the ribosome on trans-translation in vitro . Tobramycin and gentamicin, belonging to the 4,6-disubstituted class of aminoglycosides having rings I and II similar to those in the 4,5-disubstituted class, possess similar effects . Neamine, which has only rings I and II, a common structure shared by 4,5- and 4,6-disubstituted classes of aminoglycosides, was sufficient to cause an initiation shift of trans-translation . In contrast, streptomycin or hygromycin B, lacking ring I, did not cause an initiation shift . The effect of each aminoglycoside on trans-translation coincides with that on conformational change in the A site of the small subunit of the ribosome revealed by recent structural studies: paromomycin, tobramycin and geneticin which is categorized into the gentamicin subclass, but not streptomycin and hygromycin B, flip out two conserved adenine bases at 1492 and 1493 from the A site helix . The pattern of initiation shifts by paromomycin fluctuates with variation of mutations introduced into a region upstream of the initiation point. J Dent Res, 2004, 83 Spec No C, C6 - 14 The classic caries clinical trial: constraints and opportunities; Stamm JW; The history of clinical trials would include events in 1747 on board the Salisbury, a British Navy vessel at sea with 12 seamen critically ill with scurvy . Involving these 12 sailors in a study, an officer on board by the name of Lind evaluated six potential treatments for scurvy, and rapidly reached the conclusion that daily consumption of citrus fruits returned the men fit for duty in approximately six days (Bull, 1959) . The concept of experimental randomization was first developed by Sir R.A . Fisher (1925, 1926), and the method was introduced to medical research via a study of tuberculosis treatment by Amberson and co-workers (1931), who randomized 24 TB patients into two groups, one to receive the experimental therapy, the other serving as the control . Amberson et al . also incorporated the concept of blinding into their study . Sir Austin Bradford Hill codified and built on the principles of scientific experimentation developed by Fisher, and introduced the use of random numbers in the allocation of patients in the British Medical Research Council (1948) study of the effect of streptomycin in the treatment of tuberculosis (Daniels and Hill, 1952; Hill, 1952) . The first applications of clinical trial methodology for testing interventions on dental, oral, and maxillofacial diseases and conditions are more difficult to determine . For dental caries prevention, however, Chilton and Fertig (1958) and Slack and Martin (1964) were certainly among the early caries clinical trial pioneers . As clinical trials have come into the mainstream of clinical research in medicine and dentistry, a great deal of developmental work has focused on their methodological enhancement . The most successful of these efforts have come from fruitful, ongoing collaborations among clinician investigators, biostatisticians, data management specialists, biomedical ethicists, and others with an academic interest in clinical trial design and utilization . During the past 25 years, the emergence of systematic reviews and the evidence-based medicine (EBM) movement have also contributed significantly to the increasing reliance on randomized clinical trial outcomes for the advancement of better clinical practice (Richards et al., 1997; Straus and Sackett, 1998; 2002). Aust Vet J, 2004 Jun, 82(6), 370 - 4 Comparison of the efficacy of a subunit and a live streptomycin-dependent porcine pleuropneumonia vaccine; Tumamao JQ et al.; OBJECTIVE: To evaluate the efficacy of two new-generation porcine pleuropneumonia vaccines when challenged with Australian isolates of Actinobacillus pleuropneumoniae of serovars 1 and 15 . DESIGN: The Porcilis APP vaccine and an experimental streptomycin-dependent strain of A pleuropneumoniae were evaluated in a standardised pen trial . Each vaccine/challenge group consisted of 10 pigs . RESULTS: With the serovar 1 challenge, the Porcilis APP vaccine and the live vaccine, compared with the control group, gave significant protection in terms of clinical signs, lung lesions, re-isolation scores and average daily gain (ADG) postchallenge . Only the Porcilis APP vaccine provided significant protection against mortality . In the serovar 15 challenged pigs, the only significant difference detected was that the Porcilis APP vaccinated pigs had a better postchallenge ADG than the controls . None of the Porcilis APP vaccinated pigs showed signs of depression postvaccination and none were euthanased after challenge with either serovar 1 or 15 . The pigs vaccinated with the live vaccine showed obvious depression after each vaccination and a total of 3 pigs were euthanased after challenge (one with serovar 1 and two with serovar 15) . CONCLUSIONS: Both of the vaccines provided significant protection against a severe challenge with serovar 1 A pleuropneumoniae . Neither vaccine was effective against a serovar 15 A pleuropneumoniae challenge . There was evidence that the Porcilis APP vaccine did provide some protection against the serovar 15 challenge because the ADG, after challenge of pigs given this vaccine, was greater than the control pigs. Pflugers Arch, 2004 Nov, 449(2), 195 - 204 Epub 2004 Nov. Extracellular Mg(2+) blocks endothelin-1-induced contraction through the inhibition of non-selective cation channels in coronary smooth muscle; Ko EA et al.; This study investigated the effects of changing the extracellular {Mg(2+)} ({Mg(2+)}(o)) on endothelin-1 (ET-1)-induced contraction of rabbit coronary artery smooth muscle and the involvement of non-selective cation (NSC) channels in this response . Increased {Mg(2+)}(o) shifted the concentration/contraction relationship curve of ET-1 to the right . In whole-cell patch clamp recordings, ET-1 (10(-7) M) induced a long-lasting inwards current (94.7+/-7.2 pA) that was inhibited by 8 mM {Mg(2+)}(o) (45.3+/-4.4%) and NSC channel blockers (10(-3) M streptomycin and 10(-3) M La(3+)), but not by the voltage-dependent Ca(2+) channel blocker nicardipine . The current/voltage (I/V) curve was linear . Furthermore, in pressurized arteries, the ET-1-induced contraction was also inhibited by La(3+) and streptomycin, but not by nicardipine . U-73122, a selective phospholipase C (PLC) inhibitor and staurosporine and GF 109203X, which block protein kinase C (PKC), reduced ET-1-activated NSC currents by 54.2+/-5.1%, 60.3+/-5.5% and 48.5+/-2.9%, respectively . The inwards current was increased by 1-oleoyl-2-acetyl-sn-glycerol (OAG) and phorbol 12,13-dibutyrate (PDBu), which activate PKC selectively . Like transient receptor potential channel (TRPC3) currents, ET-1-activated NSC currents had a linear I/V relationship, were blocked by flufenamate and activated by a diacylglycerol analogue . These results suggest that {Mg(2+)}(o) blocks ET-1-induced contraction of coronary arteries by inhibiting NSC channels . Activation of PLC and PKC might be involved in activation of NSC channels. J Physiol, 2004 Aug 15, 559(Pt 1), 205 - 14 Epub 2004 Jul 02. Activation of Na+-H+ exchange and stretch-activated channels underlies the slow inotropic response to stretch in myocytes and muscle from the rat heart; Calaghan S et al.; We present the first direct comparison of the major candidates proposed to underlie the slow phase of the force increase seen following myocardial stretch: (i) the Na(+)-H(+) exchanger (NHE) (ii) nitric oxide (NO) and the ryanodine receptor (RyR) and (iii) the stretch-activated channel (SAC) in both single myocytes and multicellular muscle preparations from the rat heart . Ventricular myocytes were stretched by approximately 7% using carbon fibres . Papillary muscles were stretched from 88 to 98% of the length at which maximum tension is generated (L(max)) . Inhibition of NHE with HOE 642 (5 microm) significantly reduced (P < 0.05) the magnitude of the slow force response in both muscle and myocytes . Neither inhibition of phosphatidylinositol-3-OH kinase (PtdIns-3-OH kinase) with LY294002 (10 microm) nor NO synthase with L-NAME (1 mm) reduced the slow force response in muscle or myocytes (P > 0.05), and the slow response was still present in the single myocyte when the sarcoplasmic reticulum was rigorously inhibited with 1 microm ryanodine and 1 microm thapsigargin . We saw a significant reduction (P < 0.05) in the slow force response in the presence of the SAC blocker streptomycin in both muscle (80 microm) and myocytes (40 microm) . In fura 2-loaded myocytes, HOE 642 and streptomycin, but not L-NAME, ablated the stretch-induced increase in {Ca(2+)}(i) transient amplitude . Our data suggest that in the rat, under our experimental conditions, there are two mechanisms that underlie the slow inotropic response to stretch: activation of NHE; and of activation of SACs . Both these mechanisms are intrinsic to the myocyte. BMC Infect Dis . 2004 Jun 23;4(1):18. Ofloxacin plus rifampicin versus doxycycline plus rifampicin in the treatment of brucellosis: a randomized clinical trial {ISRCTN11871179}; Karabay O et al.; BACKGROUND: The combination therapies recommended by the World Health Organization for treatment of brucellosis are doxycycline plus rifampicin or doxycycline plus streptomycin . Although highly successful results have been obtained with these two regimens, relapse rates as high as 14.4% . The most effective and the least toxic chemotherapy for human brucellosis is still undetermined . The aim of the present study was to investigate the efficacy, adverse effects and cost of ofloxacin plus rifampicin therapy, and doxycycline plus rifampicin therapy and evaluate in the treatment of brucellosis . METHODS: The open trial has been carried out prospectively by the two medical centers from December 1999 to December 2001 in Duzce region Turkey . The diagnosis was based on the presence of signs and symptoms compatible with brucellosis including a positive agglutination titre (>/=1/160) and/or a positive culture . Doxycycline and rifampicin group consisted of 14 patients who were given doxycycline 200 mg/day plus rifampicin 600 mg/day during 45 days and this group Ofloxacin plus rifampicin group was consisted of 15 patients who were given ofloxacin 400 mg/day plus rifampicin 600 mg/day during 30 days . RESULTS: Regarding clinical and/or demographic characteristics no significant difference was found between two groups of patients that underwent two different therapeutic regimens . At the end of the therapy, two relapses were seen in both groups (p = 0.695) . Although duration of therapy was two weeks shorter in group treated with rifampicin plus ofloxacin, the cure rate was similar in both groups of examinees . Fever dropped more rapidly in the group that treated with rifampicin plus ofloxacin, 74 +/- 30 (ranges 48-216) vs . 106 +/- 26 (ranges 48-262) hours (p = 0.016) . CONCLUSIONS: Ofloxacin plus rifampicin therapy has advantages of shorter treatment duration and provided shorter course of fever with treatment than in doxycycline plus rifampicin therapy . However, cost of ofloxacin plus rifampicin treatment is higher than doxycycline plus rifampicin treatment . Because of the similar effects, adverse effects and relapses rates between two regimens, we still advice doxycycline plus rifampicin for the treatment of brucellosis for countries, which have limited resources. Kekkaku, 2004 May, 79(5), 341 - 8 {Leukopenia due to anti-tuberculous chemotherapy including rifampicin and isoniazid}; Nagayama N et al.; OBJECTIVES: To examine the incidence rate by age and gender of leukopenia caused by chemotherapy including rifampicin (RFP) and isoniazid (INH), and to study the relationships between the leukopenia and the hepatic side effect or other haematological disorders such as thrombocytopenia . SUBJECTS: Out of the tuberculosis patients who were admitted to our hospital in 1987-88, 1991-92, and 1996-2000, 1,525 patients (1,153 men, 372 women) were chosen for our study who had the white blood cell counts (WBC) in the peripheral blood more than 3,000/mm3 before chemotherapy, and underwent haematologic examination at least twice within 3 months after starting chemotherapy . METHODS: The definition of leukopenia was as follows: 1) WBC became less than 3,000/mm3 during chemotherapy for patients with pre-treatment WBC more than 4,000/mm3, or 2) WBC decreased more than 1,000/mm3 in patients with pre-treatment WBC between 3,000 and 4,000/mm3 . The incidence rates of leukopenia by age, gender, and regimens of chemotherapy were calculated . The case-control study was done between the control and the leukopenia groups excluding patients suffered from agranulocytosis to clarify the hematological and biochemical characteristics of the leukopenia group . The control patients were chosen in the following way . For each patient with leukopenia, a patient with the same admission year, same gender, same regimen of chemotherapy, and the nearest age was chosen as a control patient . The changes in counts of white blood cell, granulocyte, and platelet, in hemoglobin concentration, and in hepatic enzyme levels before and during chemotherapy were compared between the leukopenia and the control groups . Thrombocytopenia was defined as platelet count less than 15 x 10(4)/mm3 and hepatic dysfunction as either asparate aminotransferase (AST) higher than 31 IU/l or alanine aminotransferase (ALT) higher than 34 IU/l . RESULTS: (1) Incidence rate of leukopenia The leukopenia appeared in 36 patients (14 men, 22 women), two (one man, one woman) of whom showed agranulocytosis . The incidence rate was 1.2% (14/1,153) for men and 5.9% (22/372) for women . The incidence rate of women was higher than that of men in the age groups between 20 to 79 y.o., but no difference was seen in the age groups elder than 80 y.o . There were no differences in the incidence rate among groups treated with HRE (E: ethambutol), HRS (S: streptomycin), and HREZ (Z: pyrazinamide) . The chemotherapy was continued in 30 patients after the appearance of leukopenia, and the natural recovery from leukopenia was seen in 19 patients, while the leukopenic state lasted during the chemotherapy in the remaining 11 patients . In two patients who exhibited agranulocytosis all drugs were discontinued . In the remaining 4 patients one or more drugs were discontinued . (2) Case-control study between leukopenia (N = 34) and the control (N = 34) groups There were no differences in age, sputum culture positivity on admission, degree of roentgenographic extent of the disease, ratio of cavity formation, and quantity of daily doses between the two groups . There was also no difference between the days until leukopenia appeared after starting chemotherapy (47.6 +/- 29.5 days) in the leukopenia group, and the days until WBC count became minimum within 3 months after starting chemotherapy (41.7 +/- 21.0 days) in the control group . The negativity of tuberculin skin testing was higher in the leukopenia group {7/14 (50%)} than in the control group {1/10 (10%)}, however, the difference was statistically not significant due to rather small size of cases . Before the starting chemotherapy, the counts of WBC (7,230 +/- 1,530 vs 5,500 +/- 1,510/mm3, p < 0.001), neutrophil (5,230 +/- 1,450 vs 4,320 +/- 1,620/mm3, p < 0.05), lymphocyte (1,440 +/- 830 vs 830 +/- 440/mm3, p < 0.001) and platelet (34.9 +/- 12.2 vs 24.1 +/- 6.4 x 10(4)/mm3, p < 0.001) in the peripheral blood and the globulin level (3.71 +/- 0.61 vs 3.35 +/- 0.61 g/dl, p < 0.05) in the serum were significantly higher in the control group than in the leukopenia group . The decrements in the counts of WBC and granulocyte during chemotherapy were larger in the leukopenia group than in the control group (delta WBC: 2,880 +/- 1,530 vs 1,910 +/- 1,520/mm3, and delta Neut: 2,840 +/- 1,510 vs 1,820 +/- 1,380/mm3, p = 0.01, respectively), but the counts of lymphocyte were similar in both groups . The platelet counts also decreased in both groups, but to the mid-normal level in the control group, and to the lowest normal level in the leukopenia group, in which 15 out of 34 patients (44%) showed thrombocytopenia . The levels in the serum of hepatic enzymes such as AST, ALT, and gamma-GTP (gamma-glutamyl aminotransferase) increased during chemotherapy in the leukopenia group, while decreased in the control group, and the facts indicate that in the former not only bone marrow cells but also hepatic cells were impaired by anti-tuberculosis drugs . CONSIDERATIONS: Leukopenia may occur in the course of treatment with anti-tuberculosis drugs, but it is not necessary to stop the chemotherapy immediately, because the WBC count recovers spontaneously or remains under stable leukopenic state during chemotherapy in most cases . But when leukopenia appears, the peripheral blood counts must be checked cautiously, and the chemotherapy should be stopped if the WBC count progressively decreases . The patients who showed leukopenia due to anti-tuberculosis drugs may have had weaker natural and acquired (cell-mediated) immunologic response to tuberculosis infection, and more vulnerable bone marrow cells and hepatic cells to anti-tuberculosis drugs than the control. Methods Mol Biol, 2004, 274, 301 - 7 A simple method for chloroplast transformation in Chlamydomonas reinhardtii; Ramesh VM et al.; Photosystem (PS)I is a multi-subunit pigment-protein complex that uses light energy to transfer electrons from plastocyanin to ferredoxin . Application of genetic engineering to photo-synthetic reaction center proteins has led to a significant advancement in our understanding of primary electron transfer events and the role of the protein environment in modulating these processes . Chlamydomonas reinhardtii provides a system particularly amenable to analyze the structure-function relationship of PSI . Chlamydomonas reinhardtii is also a favorable organism for chloroplast transformation because it contains a single chloroplast and grows heterotrophically when supplemented with acetate . Chlamydomonas has served as a model organism for the development of chloroplast transformation procedures and the study of photosynthetic mutants generated using this method . Exogenous cloned cpDNA can be introduced into the chloroplast by using this biolistic gene gun method . DNA-coated tungsten or gold particles are bombarded onto cells . Upon its entry into chloroplasts, the transforming DNA is released from the particles and integrated into the chloroplast genome through homologous recombination . The most versatile chloroplast selectable marker is aminoglycoside adenyl transferase (aadA), which can be expressed in the chloroplast to confer resistance to spectinomycin or streptomycin . This chapter describes the procedures for chloroplast transformation. Acta Otolaryngol, 2004 Apr, 124(3), 258 - 61 A novel mitochondrial mutation, 1556C --> T, in a Japanese patient with streptomycin-induced tinnitus; Tanimoto H et al.; OBJECTIVE: Aminoglycoside antibiotics are associated with ototoxicity . The 1555A --> G mutation in the 12S ribosomal RNA gene of mitochondrial DNA has been considered to be associated with susceptibility to aminoglycoside antibiotics . In this study we examined a 79-year-old Japanese patient with a 45-year history of streptomycin-induced tinnitus in an attempt to find a mitochondrial mutation . MATERIALS AND METHODS: DNA was extracted from the patient's leukocytes . Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) screening for the 1555A --> G mutation was done in order to detect a mitochondrial mutation and then nucleotide sequencing analysis was performed to identify the mutation . RESULTS: PCR-RFLP screening detected the presence of a mitochondrial mutation in the patient . However, the nucleotide sequencing analysis showed that the mutation was not the 1555A --> G mutation but a novel mutation, 1556C --> T . The mutation was not found in 112 unrelated Japanese control subjects, suggesting that the mutation was specific to our patient . CONCLUSIONS: The 1556C --> T mutation may be a genetic risk factor for aminoglycoside-induced hearing impairment . Our result also suggests that patients with the 1556C --> T mutation exist among those expected to have the 1555A --> G mutation as a result of PCR-RFLP analysis. J Microbiol Methods, 2004 Jun, 57(3), 323 - 35 Multicenter evaluation of reverse line blot assay for detection of drug resistance in Mycobacterium tuberculosis clinical isolates; Mokrousov I et al.; A multicenter study was conducted with the objective to evaluate a reverse line blot (RLB) assay to detect resistance to rifampin (RIF), isoniazid (INH), streptomycin (STR), and ethambutol (EMB) in clinical isolates of Mycobacterium tuberculosis . Oligonucleotides specific for wild type and mutant (drug resistance linked) alleles of the selected codons in the genes rpoB, inhA, ahpC, rpsL, rrs, embB, were immobilized on a nylon membrane . The RLB assay conditions were optimized following analysis of DNA samples with known sequences of the targeted genes . For validation of the method at different geographical locations, the membranes were sent to seven laboratories in six countries representing the regions with high burdens of multudrug-resistant tuberculosis . The reproducibility of the assay for detection of rpoB genotypes was initially evaluated on a blinded set of twenty reference DNA samples with known allele types and overall concordant results were obtained . Further mutation analysis was performed by each laboratory on the local strains . Upon RLB analysis of 315 clinical isolates from different countries, 132 (85.2%) of 155 RIF-resistant and 28 (51.0%) of 55 EMB-resistant isolates were correctly identified, showing applicability of the assay when targeting the rpoB hot-spot region and embB306 . Mutations in the inhA and ahpC promoter regions, conferring resistance to INH, were successfully identified in respectively 16.9% and 13.2% of INH-resistant strains . Likewise, mutations in rrs513 and rpsL88 that confer resistance to STR were identified in respectively 15.1% and 10.7% of STR-resistant strains . It should be mentioned that mutation analysis of the above targets usually requires rather costly DNA sequencing to which the proposed RLB assay presents rapid and inexpensive alternative . Furthermore, the proposed method requires the same simple equipment as that used for spoligotyping and permits simultaneous analysis of up to 40 samples . This technique is a first attempt to combine different targets in a single assay for prediction of antituberculosis drugs resistance . It is open to further development as it allows easy incorporation of new probes for detection of mutations in other genes associated with resistance to second-line (e.g., fluoroquinolones) and new antituberculosis compounds. Braz J Med Biol Res, 2004 May, 37(5), 697 - 700 Epub 2004 Apr 22. BCG lymphadenopathy detected in a BCG-vaccinated infant; Barouni AS et al.; Large-scale vaccination with BCG, the live attenuated strain of Mycobacterium bovis, is being adopted around the world, although sporadic complications have occurred after the procedure . Lymphadenopathy is not uncommon especially in babies under one year (0.73% of vaccinated infants), but the swelling subsides within 2 months in most cases, with no medical or surgical treatment . Brazil adopted BCG vaccination program earlier in the seventies and by 1995 more than 96% of the infant population received this immunization . We report here the occurrence of lymphadenopathy in a two-year-old child vaccinated with the Brazilian BCG strain . The diagnosis was made using a lymph node biopsy and intestinal aspirates that yielded a positive mycobacterial culture . The isolate was resistant to isoniazid, rifampicin, pyrazinamide and thiophen-2-carbonic acid hydrazide, sensitive to streptomycin, ethambutol, and p-nitrobenzoic acid, and reacted positively to cyclo-serine and negatively to niacin . The pncA gene involved in bacterial activation of pyrazinamide contains in M . bovis a point mutation that renders pyrazinamidase unable to catalyze drug activation . Therefore, this polymorphism is a good option for developing methods to differentiate M . bovis and M . tuberculosis . Taking advantage of this difference we further analyzed the isolates by single-stranded conformation polymorphism electrophoresis of DNA following PCR of the pncA gene . The isolate identity was confirmed by RFLP electrophoretic analysis of the amplified fragment following Eco065I digestion, which selectively cleaves M . tuberculosis DNA . From this result it is proposed that RFLP of pncA gene represents an alternative for differential diagnosis of M . bovis. Circ Res, 2004 May 28, 94(10), 1392 - 8 Epub 2004 Apr 22. Functional relevance of the stretch-dependent slow force response in failing human myocardium; von Lewinski D et al.; Stretch induces immediate and delayed inotropic effects in mammalian myocardium via distinct mechanosensitive pathways, but these effects are poorly characterized in human cardiac muscle . We tested the effects of stretch on immediate and delayed force response in failing human myocardium . Experiments were performed in muscle strips from 52 failing human hearts (37 degrees C, 1 Hz, bicarbonate buffer) . Muscles were stretched from 88% of optimal length to 98% of optimal length . The resulting immediate and delayed (ie, slow force response {SFR}) increases in twitch force were assessed without and after blockade of the sarcoplasmic reticulum (SR; cyclopiazonic acid and ryanodine), stretch-activated ion channels (SACs; gadolinium, streptomycin), L-type Ca2+-channels (diltiazem), angiotensin II type-1 (AT1) receptors (candesartan), endothelin (ET) receptors (PD145065 or BQ123), Na+/H+ exchange (NHE1; HOE642), or reverse-mode Na+/Ca+ exchange (NCX; KB-R7493) . We also tested the effects of stretch on SR Ca2+ load (rapid cooling contractures {RCCs}) and intracellular pH (in BCECF-loaded trabeculae) . Stretch induced an immediate (<10 beats), followed by a slow (5 to 10 minutes), force response . Twitch force increased to 232+/-6% of prestretch value during the immediate phase, followed by a further increase to 279+/-8% during the SFR . RCC amplitude significantly increased, but pHi did not change during SFR . Inhibition of SACs, L-type Ca2+ channels, AT1 receptors, or ET receptors did not affect the stretch-dependent immediate or SFR . In contrast, the SFR was reduced by NHE1 inhibition and almost completely abolished by reverse-mode NCX inhibition or blockade of sarcoplasmic reticulum function . The data demonstrate the existence of a functionally relevant, SR-Ca2+-dependent SFR in failing human myocardium, which partly depends on NHE1 and reverse-mode NCX activation. J AOAC Int, 2004 Jan-Feb, 87(1), 39 - 44 Evaluation of analyte stability and method ruggedness in the determination of streptomycin residues in honey by liquid chromatography with post-column derivatization; Pang GF et al.; This study demonstrated that streptomycin in honey is quite stable, and the results showed no obvious differences for 3 samples containing incurred analyte during continuous testing for 4 months . Fifteen laboratories evaluated method performance at 4 fortification levels ranging from 0.010 to 0.100 mg/kg; the recoveries ranged from 73.7 to 78.5%, the reproducibility relative standard deviations ranged from 5.76 to 15.85%, and the repeatability relative standard deviations ranged from 1.64 to 3.80% . In 1999-2002, the method was used to determine streptomycin residues in 5106 lots of honey samples from >20 provinces all over China . All of the honey samples were found to be in conformity with the requirements of customs clearance for exports to Europe, the United States, and Japan . The continuous 4-year quality analysis also found that C18 solid-phase extraction cartridges should be standardized to ensure that the analytical results are accurate when different lots of cartridges are used. Avian Dis, 2004 Jan-Mar, 48(1), 19 - 25 Attenuation of an avian pathogenic Escherichia coli strain due to a mutation in the rpsL gene; Amoako KK et al.; The diseases caused by pathogenic Escherichia coli constitute a major economic loss to the poultry industry . The development of a live oral E . coli vaccine to prevent or reduce diseases in poultry had been the objective of our work . Four spontaneous streptomycin-dependent (str-dependent) mutants were generated from a virulent avian strain that contains a mutation in the fur region of the chromosome . Genetic analysis of the mutants indicated that the str-dependent phenotype was due to a base change of C --> T at base 272 in the rpsL gene . The mutants were tested for attenuation using the day-old chick model . Day-old birds, in groups of 20, were either challenged with 10(6) colony-forming units (CFU) of the str-dependent mutant, the parent strain (containing the fur mutation), or the wild-type strain without the fur mutation . The parent strain and the wild-type strain were highly virulent, and 80% or more of the birds died . None of the birds challenged with the str-dependent mutants died, indicating attenuation of the mutants . The protective effect of the mutant as a live vaccine against the challenge with 10(6) CFU of the wild-type strain EC317 was investigated . Vaccination by both aerosol (day 1) and oral (days 14 and 28) routes using 10(8) CFU of the str-dependent mutant (EC1598) had no effect on the occurrence of cellulitis in the birds . Two vaccinations given as aerosol on day 1 and given orally on day 14 also had no significant effect on the occurrence of systemic lesions . Three immunizations on days 1, 14, and 28 resulted in a significant reduction in the number of birds with systemic lesions . Antibody titers prior to challenge were not predictive of outcome of challenge. Plant Cell Physiol, 2004 Mar, 45(3), 333 - 9 High-frequency gene replacement in cyanobacteria using a heterologous rps12 gene; Takahama K et al.; Multiple targeted gene replacements are often required for functional analyses of cyanobacterial genomes . For this purpose, we previously devised a simple genetic method, termed rps12-mediated gene replacement, in a cyanobacterium Synechococcus elongatus PCC 7942 for construction of mutants free from drug resistance markers . Here, we improved the method by employing a heterologous rps12 gene encoding a ribosomal protein S12 from Synechocystis sp . PCC 6803 . Dominant streptomycin-sensitive phenotype of the Synechocystis rps12 gene was manifested only when it was expressed under the strong promoter of psbAI gene in S . elongatus PCC 7942 bearing a streptomycin-resistant rps12 allele . Transformation of the rps12 heteroallelic strains with non-replicating template plasmids permitted the selection of recombinants with gene replacement at frequencies up to 50% among streptomycin-resistant progeny. Plant Cell Rep, 2004 Feb, 22(7), 490 - 6 Epub 2003 Sep 30. Development of a non-lethal selection system by using the aadA marker gene for efficient recovery of transgenic rice (Oryza sativa L.); Oreifig AS et al.; The application of aminoglycoside-3"-adenyltransferase ( aadA) gene-mediated streptomycin resistance for non-lethal selection of transgenic rice resulted in plant regeneration frequencies under selection pressure as high as those in non-transformed controls without selection . Since streptomycin does not kill non-transgenic cells, and allows plant regeneration from them, a selection procedure was developed that made the visual identification of transgenic calli and regenerants possible . For callus-level selection, a vital pH indicator-Chlorophenol Red-was applied together with streptomycin, making use of the phenomenon that fast-growing cell lines lower the pH in the culture medium . Transgenic plants were selected according to their main distinctive features; their green colour (photomixotrophic assimilation), and more intense growth . At the same time, non-transgenic regenerants were bleached (heterotrophic assimilation), and growth was retarded in the presence of streptomycin and sucrose . The final efficiency of genetic transformation based on streptomycin resistance was found to be double that of transformations where the selective agent was l-phosphinothricin, and nearly three times more compared to transformations resulting in hygromycin-resistant regenerants . To the best of our knowledge, this is the first report on producing nuclear transformed rice plants by using a non-lethal selection strategy based on the chimaeric aadA gene. J Bacteriol, 2004 Apr, 186(7), 2206 - 11 A single target is sufficient to account for the biological effects of the A-factor receptor protein of Streptomyces griseus; Kato JY et al.; In the model of the A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) regulatory cascade in Streptomyces griseus, A-factor binds ArpA, the A-factor receptor protein, that has bound to the adpA promoter and dissociates it from the DNA, thus inducing the transcription of adpA . AdpA switches on the transcription of a number of genes required for secondary metabolism and morphological differentiation, forming an AdpA regulon . Consistent with this model, arpA null mutants produced streptomycin and a yellow pigment in larger amounts and formed aerial hyphae from an earlier growth stage than the wild-type strain . On the other hand, mutant MK2, expressing a mutant ArpA (Trp119Ala), neither produced secondary metabolites nor formed aerial hyphae, because this A-factor-insensitive mutant ArpA always bound to and repressed the adpA promoter due to the amino acid replacement of Trp-119 with Ala . Introduction of adpA under the control of a foreign promoter into mutant MK2 restored all of the phenotypes that we could observe, which suggests that the only significant target of ArpA is adpA . In contrast to other gamma-butyrolactone regulatory systems, disruption of arpA had no effect on A-factor production, indicating that ArpA does not regulate A-factor biosynthesis . Instead, A-factor production was found to be repressed by AdpA in a two-step regulatory feedback loop. Theor Appl Genet, 2004 Jul, 109(2), 333 - 41 Epub 2004 Mar 10. Mapping of transposable element Dissociation inserts in Brassica oleracea following plant regeneration from streptomycin selection of callus; McKenzie N et al.; To investigate the potential of heterologous transposons as a gene-tagging system in broccoli (Brassica oleracea var . italica), we have introduced a Dissociation ( Ds)-based two-element transposon system . Ds has been cloned into a 35S-SPT excision-marker system, with transposition being driven by an independent 35S-transposase gene construct . In three successive selfed generations of plants, there was no evidence of germinal-excision events . In a previous study, we overcame this apparent inability to produce B . oleracea plants with germinal excisions by performing a novel tissue-culture technique to select for fully green shoots from seed with somatic excision events . The results showed a very high efficiency of regeneration of fully green plants (up to 65%), and molecular analysis showed that the plants contained the equivalent of a germinal-excision event . In this study, we followed the previous work by using inverse and nested PCR to generate probes of flanking genomic DNA adjacent to independently reinserted Ds elements, and these were hybridised to DNA from a double-haploid mapping population of B . oleracea . Seventeen Ds insertions and the original Ds T-DNA site have been localised, and these are spread over six (out of nine) linkage groups . Distribution of inserts show that 15 were found on a different linkage group to the original 'launch' site, and of these 11 were found to be clustered on two separate groups . Previous studies in other plant species have found that germinal excision of Ds predominantly moves to sites linked close to the donor site . However, this study shows a potential to produce plants with Ds insertion scattered over many unlinked sites. Eur J Biochem, 2004 Mar, 271(6), 1127 - 34 Effects of Escherichia coli ribosomal protein S12 mutations on cell-free protein synthesis; Chumpolkulwong N et al.; We examined the effects of Escherichia coli ribosomal protein S12 mutations on the efficiency of cell-free protein synthesis . By screening 150 spontaneous streptomycin-resistant isolates from E . coli BL21, we successfully obtained seven mutants of the S12 protein, including two streptomycin-dependent mutants . The mutations occurred at Lys42, Lys87, Pro90 and Gly91 of the 30S ribosomal protein S12 . We prepared S30 extracts from mutant cells harvested in the mid-log phase . Their protein synthesis activities were compared by measuring the yields of the active chloramphenicol acetyltransferase . Higher protein production (1.3-fold) than the wild-type was observed with the mutant that replaced Lys42 with Thr (K42T) . The K42R, K42N, and K42I strains showed lower activities, while the other mutant strains with Lys87, Pro90 and Pro91 did not show any significant difference from the wild-type . We also assessed the frequency of Leu misincorporation in poly(U)-dependent poly(Phe) synthesis . In this assay system, almost all mutants showed higher accuracy and lower activity than the wild-type . However, K42T offered higher activity, in addition to high accuracy . Furthermore, when 14 mouse cDNA sequences were used as test templates, the protein yields of nine templates in the K42T system were 1.2-2 times higher than that of the wild-type. J Clin Microbiol, 2004 Mar, 42(3), 1109 - 14 Evaluation of the fully automated BACTEC MGIT 960 system for testing susceptibility of Mycobacterium tuberculosis to pyrazinamide, streptomycin, isoniazid, rifampin, and ethambutol and comparison with the radiometric BACTEC 460TB method; Scarparo C et al.; The performance of the fully automated BACTEC MGIT 960 (M960) system for the testing of Mycobacterium tuberculosis susceptibility to streptomycin (SM), isoniazid (INH), rifampin (RMP), ethambutol (EMB), and pyrazinamide (PZA) was evaluated with 100 clinical isolates and compared to that of the radiometric BACTEC 460TB (B460) system . The agar proportion method and the B460 system were used as reference methods to resolve the discordant results for SM, INH, RMP, and EMB (a combination known as SIRE) and PZA, respectively . The overall agreements were 96.3% for SIRE and 92% for PZA . For SIRE, a total of 26 discrepancies were found and were resolved in favor of the M960 system in 8 cases and in favor of the B460 system in 18 cases . The M960 system produced 8 very major errors (VME) and 10 major errors (ME), while the B460 system showed 4 VME and 4 ME . No statistically significant differences were found . Both systems exhibited excellent performance, but a higher number of VME was observed with the M960 system at the critical concentrations of EMB and SM . For PZA, a total of eight discrepancies were observed and were resolved in favor of the M960 system in one case and in favor of the B460 system in seven cases; no statistically significant differences were found . The M960 system showed four VME and three ME . The mean times to report overall PZA results and resistant results were 8.2 and 9.8 days, respectively, for the M960 system and 7.4 and 8.1 days, respectively, for the B460 system . Statistically significant differences were found . The mean times to report SIRE results were 8.3 days for the M960 system and 8.2 days for the B460 system . No statistically significant differences were found . Twelve strains tested for SIRE susceptibility and seven strains tested for PZA susceptibility had been reprocessed because of contamination . In conclusion, the M960 system can represent a valid alternative to the B460 for M . tuberculosis susceptibility testing; however, the frequent contamination of the tests needs to be improved. J Clin Microbiol, 2004 Mar, 42(3), 1030 - 4 Multicenter evaluation of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis; Bemer P et al.; The reliability of the MB/BACT system for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide, rifampin, isoniazid, streptomycin, and ethambutol was compared to the BACTEC 460TB system . The proportion method was used to resolve discrepant results by an independent arbiter . Two interpretative methods were used, with an undiluted control (direct control) and a diluted control (10(-1) control) . As no significant difference was observed between the two controls, the method with the direct control was adopted as the most accurate one . One hundred sixty-six strains were tested, with an overall agreement of 98.3% . After resolution of the 18 discrepant results by the proportion method, the sensitivity and specificity of the MB/BACT system were 100% for rifampin, isoniazid, and pyrazinamide . For ethambutol, sensitivity was 92.3% at the critical concentration and 33% at the high concentration, and specificity was 100% at both concentrations . For streptomycin, sensitivity was 100% at the critical concentration and 80% at the high concentration, and specificity was 98.6% at the critical concentration and 100% at the high concentration . The rifampin, isoniazid, streptomycin, and ethambutol susceptibility test results were obtained in 6.6 days with the MB/BACT versus 5 days with the BACTEC 460TB . The pyrazinamide susceptibility test results were obtained in 7.8 days with the MB/BACT, versus 6.7 days with the BACTEC 460TB . These data demonstrate that the fully automated MB/BACT system is a very reliable method for rapid susceptibility testing of M . tuberculosis against rifampin, isoniazid, and pyrazinamide . Sensitivity results have to be improved for ethambutol and streptomycin, especially at the high concentration. Microb Drug Resist, 2003 Winter, 9(4), 361 - 6 Primary drug resistance and molecular epidemiology of Mycobacterium tuberculosis isolates from patients in a population with high tuberculosis incidence in Turkey; Durmaz R et al.; To determine the rate of primary drug resistance and compare the fingerprint pattern diversity of the resistant and sensitive Mycobacterium tuberculosis isolates, antituberculosis susceptibility testing and restriction fragment length polymorphism (RFLP) analysis were performed on 88 M . tuberculosis isolates of the patients who were diagnosed as new tuberculosis cases in 2000 . Primary resistance to isoniazid, rifampicin, ethambutol, and streptomycin were determined by the BACTEC method . IS6110 and pTBN12 were used as molecular markers . The frequency of resistance to at least one drug was 32.95%, whereas 10.23% of the isolates were resistant to more than one drug . Single-drug resistance to isoniazid, streptomycin, ethambutol, and rifampicin was found in 9 (10.22%), 7 (7.95%), 4 (4.54%), and 0 (0.0%) strains, respectively . Two M . tuberculosis strains (2.26%) showed multiple drug resistance . The combination of two fingerprinting procedures on a total of 88 isolates identified 58 (65.9%) strains as unique and clustered 30 strains in 11 clusters (clustering = 34.1%) . The clustering rate for resistant and sensitive isolates was 13.8% and 40.1%, respectively . In conclusion; drug susceptibility testing showed that the majority of the drug-resistant infections involved either isoniazid or streptomycin alone . In addition to the high tuberculosis incidence, elevated primary drug resistance and high clustering rate indicate problems in the present control programs . New control strategies supported by molecular typing might be more effective to reduce tuberculosis. Hinyokika Kiyo, 2003 Dec, 49(12), 761 - 4 {A case of suspected tuberculous retroperitoneal abscess effectively cured using sclerotherapy with minocycline}; Yamamoto T et al.; A twenty-five-year-old female was admitted with lower right abdominal pain, right coxalgia and an inability to extend her right inferior limb . She had a history of tuberculosis pleurisy two years before . Abdominal ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI) demonstrated a right retroperitoneal mass which was suspected to be an abscess or tumor . Percutaneous aspiration of the mass was followed by the administration (p.o.) of antituberculosis drugs (pyrazinamide, ethanbutol, isoniazide, refampicin) . One month after initial drainage, the tube was removed but intra-cystic fluid collection was still visible a month later using CT and MRI . Therefore, a second percutaneous aspiration was followed by the instillation of streptomycin and minocycline hydrochloride . Six months after employing this therapy, no fluid collection was found. Am J Hematol, 2004 Mar, 75(3), 139 - 41 Immune thrombocytopenia attributed to brucellosis and other mechanisms of Brucella-induced thrombocytopenia; Pappas G et al.; Thrombocytopenia often complicates the course of acute brucellosis, mainly due to bone marrow suppression or hypersplenism . Immune thrombocytopenia is also reported in brucellosis, resulting usually in massive thrombocytopenia, purpura, and spontaneous hemorrhage . We describe a case of acute brucellosis in an 85-year old woman, who presented with fever, purpuric skin lesions, anemia, and rhinorrhagia . The absolute platelet count was 1000/microL . Direct and indirect Coombs tests were positive, and a cold-agglutinin was detected . The patient was diagnosed as suffering from brucellosis on the basis of a strongly positive serologic reaction and was treated with doxycycline, streptomycin, and a short course of corticosteroids, with a rapid rise in platelet number . Infect Immun, 2004 Mar, 72(3), 1666 - 76 Glycolytic and gluconeogenic growth of Escherichia coli O157:H7 (EDL933) and E . coli K-12 (MG1655) in the mouse intestine; Miranda RL et al.; Escherichia coli EDL933, an O157:H7 strain, is known to colonize the streptomycin-treated CD-1 mouse intestine by growing in intestinal mucus (E . A . Wadolkowski, J . A . Burris, and A . D . O'Brien, Infect . Immun . 58:2438-2445, 1990), but what nutrients and metabolic pathways are employed during colonization has not been determined . In this study, when the wild-type EDL933 strain was fed to mice along with an EDL933 DeltappsA DeltapckA mutant, which is unable to utilize tricarboxylic acid cycle intermediates and gluconeogenic substrates for growth, both strains colonized the mouse intestine equally well . Therefore, EDL933 utilizes a glycolytic substrate(s) for both initial growth and maintenance when it is the only E . coli strain fed to the mice . However, in the presence of large numbers of MG1655, a K-12 strain, it is shown that EDL933 utilizes a glycolytic substrate(s) for initial growth in the mouse intestine but appears to utilize both glycolytic and gluconeogenic substrates in an attempt to maintain colonization . It is further shown that MG1655 predominantly utilizes glycolytic substrates for growth in the mouse intestine whether growing in the presence or absence of large numbers of EDL933 . Data are presented showing that although small numbers of EDL933 grow to large numbers in the intestine in the presence of large numbers of MG1655 when both strains are fed to mice simultaneously, precolonization with MG1655 affords protection against subsequent colonization by EDL933 . Moreover, in mice that are precolonized with EDL933, small numbers of MG1655 are able to grow rapidly in the intestine and EDL933 is eliminated . In situ hybridization experiments using E . coli-specific rRNA probes showed that while MG1655 is found only in mucus, EDL933 is found both in mucus and closely associated with intestinal epithelial cells . The data are discussed with respect to competition for nutrients and to the protection that some intestinal commensal E . coli strains might afford against infection by O157:H7 strains. Mol Genet Genomics, 2004 Apr, 271(3), 317 - 24 Epub 2004 Feb 14. The novel mutation K87E in ribosomal protein S12 enhances protein synthesis activity during the late growth phase in Escherichia coli; Hosaka T et al.; Resistance to streptomycin in bacterial cells often results from a mutation in the rpsL gene that encodes the ribosomal protein S12 . We found that a particular rpsL mutation (K87E), newly identified in Escherichia coli, causes aberrant protein synthesis activity late in the growth phase . While protein synthesis decreased with age in cells in the wild-type strain, it was sustained at a high level in the mutant, as determined using living cells . This was confirmed using an in vitro protein synthesis system with poly(U) and natural mRNAs (GFP mRNA and CAT mRNA) . Other classical rpsL mutations (K42N and K42T) tested did not show such an effect, indicating that this novel characteristic is typical of ribosomes bearing the K87E mutant form of S12, although the K87E mutation conferred the streptomycin resistance and error-restrictive phenotypes also seen with the K42N and K42T mutations . The K87E (but not K42N or K42T) mutant ribosomes exhibited increased stability of the 70S complex in the presence of low concentrations of magnesium . We propose that the aberrant activation of protein synthesis at the late growth phase is caused by the increased stability of the ribosome. Lik Sprava, 2003 Dec, (8), 16 - 20 {M . tuberculosis drug resistance and occurrence in patients with pulmonary tuberculosis}; Novozhylova IO; The paper presents the results of M . tuberculosis drug resistance analysis . The material was phlegm obtained from 1060 patients treated during 1994-2001 y.y . The tendency of rising of M . tuberculosis drug resistance rate and polyresistance has been established . Unfortunately, a great deal of patients were undercovered with drug resistance M . tuberculosis which can massively grow on cultural medium with peak concentration of streptomycin, kanamycin, rifampicin and PAS(A), as stated above it necessitates to modernize the inspection system directed to determine M . tuberculosis drug resistance. Nihon Kokyuki Gakkai Zasshi, 2004 Jan, 42(1), 103 - 7 {A case of pulmonary Mycobacterium gordonae infection with pleural effusion}; Saeki S et al.; A 65-year-old woman, treated with prednisolone (5 mg daily) for rheumatoid arthritis, visited our hospital because of right chest pain . Chest CT showed small nodular shadows in the right lung accompanied with right pleural effusion . A pulmonary Mycobacterium gordonae infection was diagnosed, since M . gordonae was identified twice from her sputum . She was treated with rifampicin, ethambutol and streptomycin for two months, and then streptomycin was replaced with clarithromycin . Three months after the initial treatment, M . gordonae was eradicated from her sputum . Pleural puncture revealed bloody, exudative, lymphocytotic pleural effusion, but no malignant cells were identified . Although pathological diagnosis by thoracoscopic pleural biopsy could not be performed, it is likely that the pleural effusion was associated with the pulmonary M . gordonae infection in the present case. Medicina (Kaunas), 2004, 40(1), 42 - 5 {Clinical forms of new cases of tuberculosis at Kaunas Romainiai Tuberculosis Hospital in 1998-2001}; Naudziunas A et al.; Totally 1427 patients with tuberculosis were investigated in Kaunas Romainiai Tuberculosis Hospital . All patients belonged to the first category (new cases positive for mycobacterium tuberculosis, or severe tuberculosis) . Infiltrated and disseminated tuberculosis were the most frequent clinical forms . The frequency of infiltrated tuberculosis ranged from 57.36 to 68.8%, and disseminated tuberculosis ranged from 18.52 to 30.36% . The most frequent complications were bleeding from the lungs and chronic cor pulmonale . The resistance to isoniazid, rifampicin and streptomycin was investigated . Multi-drug resistant tuberculosis did not exceed 1% . We suggest that so few cases of multi-drug resistant tuberculosis are due to implementation of WHO treatment standards. J Microbiol Methods, 2004 Feb, 56(2), 291 - 4 Evaluation of the fully automated Bactec MGIT 960 system for the susceptibility testing of Mycobacterium tuberculosis to first-line drugs: a multicenter study; Kontos F et al.; The accuracy of the Bactec MGIT 960 system for susceptibility testing of 177 clinical isolates of Mycobacterium tuberculosis to first line drugs (isoniazid, rifampicin, ethambutol and streptomycin) was compared with the agar reference method . The sensitivity, the ability to detect resistance, of the MGIT system was 100%, while the specificity, the ability to detect susceptibility, ranged from 98.6% to 100% for all drugs tested. Pneumologie, 2004 Jan, 58(1), 23 - 7 {Pleuritis tuberculosa - therapeutic value of repeated chest tapping}; Hoheisel G et al.; Tuberculous pleuritis tends to develop fibrosis to a high degree . The use of corticosteroids enhances the absorption of pleural effusions, the residual pleural thickening, however, remains unaffected . Whether repeated chest tapping in patients with persistent effusions in addition to antituberculous therapy favourably influences the outcome is not known . Therefore, patients with tuberculous pleuritis were examined in a prospective, randomized study . After confirmation of the diagnosis patients were randomized in group A with antituberculous treatment (Isoniazid, Rifampicin, Pyrazinamid, and Streptomycin) alone or in group B with additional pleural tapping for four weeks (phase I) . In phase II patients in both groups with persistent effusions received oral prednisolone (0,75 mg/kg body weight) tapered over four weeks . The extent of pleural effusions was determined by chest X-ray . Roentgenological changes were evaluated at the end of the observation period . Lung function tests b