AIMS This study was designed to evaluate the prevalence of bacterial colonization of generator pockets in implantable cardioverter defibrillator (ICD) patients without signs of infection and to analyse the impact of bacterial colonization on the incidence of device infection during follow-up. METHODS AND RESULTS In 122 ICD patients undergoing generator replacement or surgical lead revision between January 2006 and July 2008, microbiological cultures of generator pockets and extracted leads were consecutively obtained. Patients with clinical evidence of a device infection were excluded. Positive cultures from the generator pocket and leads were found in 40 (33%) patients. The most common bacteria isolated were coagulase negative staphylococci (68%). During a median follow-up time of 203 days after the revision device infection occurred in three [7.5%, confidence interval (CI) 1.6-20.4%] patients with a positive culture vs. two (2.4%, CI 0.3-8.5%) patients with a negative culture (P = 0.33). Time from revision to infection was 108 +/- 73 days in patients with positive culture vs. 60 +/- 39 days in patients with negative culture (P = 0.50). CONCLUSION A third of ICD patients undergoing generator replacement or lead revision have an asymptomatic bacterial colonization of generator pockets. After revision 7.5% of these patients develop a device infection with the same species of microorganism.

Allograft bone is commonly used in reconstructive orthopaedic surgery and needs to be assessed for bioburden before transplant. The Microbiology Department of the South Eastern Area Laboratory Services (SEALS), located at the St. George Hospital, Sydney, has provided this service to the New South Wales (NSW) Bone Bank. This study reviewed the organisms isolated from femoral head allografts of living donors from the NSW Bone Bank over a 7-year period. It was found that growth was reported from 4.9% of samples with the predominant organism being coagulase-negative staphylococci. This review will focus on the micro-organisms isolated, the interaction of the laboratory with the bone bank, the relevance of the bioburden assessment in the overall quality process and patient safety.

Transport media should preserve the viability and stability of microorganisms in clinical specimens. In this study, the Port-A-Cul transport system and the Copan transport system without charcoal, both designed to preserve anaerobes, were evaluated. Dacron swabs were inoculated with two combinations of facultative and anaerobic organisms typically found in vaginal swab samples. Combination I contained Candida albicans, Escherichia coli, Enterococcus spp., group B streptococci, Lactobacillus crispatus, and Staphylococcus aureus. Combination II contained Lactobacillus iners, Peptoniphilus asaccharolyticus, Mycoplasma hominis, Prevotella bivia, Prevotella corporis, Porphyromonas asaccharolytica, Mobiluncus curtisii, Peptostreptococcus anaerobius, and Gardnerella vaginalis. Duplicate swabs were placed into the two transporters and held for 24, 48, 72, and 96 h at 4 and 24 degrees C. Both transporters maintained the viability of organisms better at 4 degrees C than at 24 degrees C. Prevotella bivia and Prevotella corporis had a loss of viability in both transporters at both temperatures. However, at 24 degrees C, there was a significantly greater loss of viability for Mycoplasma hominis, Prevotella bivia, Prevotella corporis, and Peptoniphilus asaccharolyticus when the organisms were stored in Copan transport medium than when they were stored in Port-A-Cul transport medium for 96 h (P < 0.002). Some organisms proliferated in the transport media, but when transporters were held at 24 degrees C for 96 h, a significantly greater increase in the concentrations of group B streptococci and Candida albicans, Escherichia coli, and Enterococcus spp. organisms in Copan medium than in Port-A-Cul medium was observed (P < 0.002). At room temperature, the Port-A-Cul system is superior to the Copan system with respect to the preservation of fastidious microorganisms and the prevention of the proliferation of facultative organisms.

Eno powder (GlaxoSmithKline), an antacid preparation readily available over the counter, was used instead of a CO(2) generator for the growth of 15 strains of Neisseria gonorrhoeae obtained from men with urethritis. Due to its easy accessibility and low cost, Eno powder can be useful in developing countries for transporting clinical specimens from resource-poor peripheral labs to reference laboratories.

In the present study, we followed the CLSI procedure M40-A to evaluate three specimen transport systems [the new BD CultureSwab MaxV(+), the new Remel BactiSwab, and the Medical Wire & Equipment Transwab] for the survival of fastidious and nonfastidious organisms for 0, 6, 24, and 48 h at room temperature. BD CultureSwab MaxV(+) outperformed the other two swabs for the recovery of the three fastidious organisms, Haemophilus influenzae, Neisseria gonorrhoeae, and Neisseria meningitidis for up to 48 h. Indeed, BD CultureSwab MaxV(+) maintained a constant number of viable H. influenzae and N. meningitidis for up to 48 h, and only a 2 log reduction was noted for N. gonorrhoeae, fulfilling the requirements of M40-A guidelines. However, unlike Remel BactiSwab and the Medical Wire & Equipment Transwab, which fulfilled the M40-A requirements for maintaining the viability of Streptococcus pneumoniae, BD CultureSwab MaxV(+) could not maintain the viability of S. pneumoniae reference or clinical strains past 6 h. Excellent overall sensitivity (98%)(95% confidence interval, 89.5 to 99.7) was observed when the BD CultureSwab MaxV(+) rectal swabs were compared to the "gold standard" stool cultures. Thus, the BD CultureSwab MaxV(+) rectal swab can be used when investigating gastrointestinal bacterial outbreaks or when health care providers face difficulties in obtaining stool samples, particularly from children.

Despite limitations of swab transport systems (STS), many clinical samples and bacterial isolates are sent on these devices. We evaluated the performance of 4 commercial STS: M40 Transystemtrade mark (S1, Copan Italia, Bovezzo, Italy), UNI-TERtrade mark (S2, MEUS, Piove di Sacco, Italy), Euromedtrade mark (S3, LAB SERVICE, Surbo, Italy), and Eurotubotrade mark (S4, Deltalab, Rubí, Spain). Survival of ATCC and clinical strains with characterized resistance mechanisms, stored at room temperature and using the roll-plate method (NCCLS M40-A) after holding times of 0, 6, 24, 48, and 72 h, and 7 days, was assessed. After 24 h, only S1 was able to maintain Streptococcus pneumoniae (ATCC 6305), Haemophilus influenzae (10211), and Neisseria meningitidis (13090) viability with a percentage range of recovery with respect to the baseline count at zero time of 104-184%. Neisseria gonorrhoeae (43069) was uncultivable after 6 h with all STS except S1 (46% recovery). Eighteen percent recovery after 24 h and 7% after 6 h was observed for Fusobacterium nucleatum (25586) and Peptostreptococcus anaerobius (27337), respectively, but only with S1. Resistance mechanisms in clinical isolates do not affect survival with any of the STS.

BACKGROUND The isolation of a pathogen is vital in the diagnosis and treatment of a device infection. A swab culture, despite poor sensitivity, is the most common method used in specimen collection. OBJECTIVE To determine the relative value of swab and tissue specimen cultures in patients with implantable cardiac pacemakers and defibrillators. DESIGN Prospective patient cohort study. SETTING A 1,000-bed tertiary referral center in Cleveland, Ohio. PATIENTS Consecutive patients with implantable cardiac pacemaker or defibrillator presenting for lead extraction from October 1, 2000 to March 31, 2001. METHODS Tissue and swab cultures were prospectively collected during pacemaker and implantable defibrillator surgeries that required lead extraction. Clinical manifestations, microbiology, and echocardiographic data were recorded in patients with and without a clinical diagnosis of device system infection. RESULTS Seventy-one patients with implantable pacemaker (n = 49, 69%), implantable defibrillator (n = 18, 25%), or both devices (n = 4, 6%) requiring lead extraction had pocket swab and tissue cultures for analysis. Infection was evident clinically in 35 (49%) of the patients and absent in the remainder. The most common bacteria isolated were coagulase-negative Staphylococcus (37%) and Staphylococcus aureus (10%). Patients with clinical infection had positive cultures more frequently (P = 0.002) by pocket tissue culture (n = 24, 69%) than by swab culture (n = 11, 31%). However, patients without clinical infections had positive cultures at similar rates by pocket tissue culture (n = 10, 28%) and by swab culture (n = 8, 22%; P = 0.48). Patients without clinical infection were not treated with other than perioperative antibiotics, and did not develop clinical infections. CONCLUSION Pocket tissue cultures are more effective than pocket swab cultures for the isolation and identification of the infectious pathogens in cardiac device infections. Positive cultures by pocket swab or tissue cultures in the absence of clinical signs and symptoms of infection does not imply infection or the need for specific therapy.

Clostridium perfringens is a pathogen of great concern in veterinary medicine, because it causes enteric diseases and different types of toxaemias in domesticated animals. It is important that bacteria in tissue samples, which have been collected in the field, survive and for the classification of C. perfringens into the correct toxin group, it is crucial that plasmid-borne genes are not lost during transportation or in the diagnostic laboratory. The objectives of this study were to investigate the survival of C. perfringens in a simulated transport of field samples and to determine the stability of the plasmid-borne toxin genes cpb1 and etx after storage at room temperature and at 4 degrees C. Stability of the plasmid-borne genes cpb1 and etx of C. perfringens CCUG 2035, and cpb2 from C. perfringens CIP 106526, JF 2255 and 6 field isolates in aerobic atmosphere was also studied. Survival of C. perfringens was similar in all experiments. The cpbl and etx genes were detected in all isolates from samples stored either at room temperature or at 4 degrees C for 24-44 h. Repeated aerobic treatment of C. perfringens CCUG 2035 and CIP 106526 did not result in the loss of the plasmid-borne genes cpb1, cpb2 or etx. Plasmid-borne genes in C. perfringens were found to be more stable than generally reported. Therefore, C. perfringens toxinotyping by PCR can be performed reliably, as the risk of plasmid loss seems to be a minor problem.

PURPOSE To compare the microbiological yield of corneal ulcer cultures established by direct inoculation of culture media vs indirect inoculation by means of transport medium (Amies without charcoal). DESIGN Single masked, prospective clinical trial. METHODS Scrapings were obtained for Gram and potassium hydroxide (KOH) stains from eyes with presumed infectious keratitis and cultured by direct plating onto standard media. Samples were also held in transport media (Amies without charcoal) at room temperature and then plated after 4 and 24 hours. Yields from direct plating vs cultures by means of transport media were compared. RESULTS Of 100 consecutive eyes examined with presumed infectious keratitis, Gram or KOH stain revealed a bacterial or fungal agent in 69 cases (69%). Of these, 26 were bacterial and 43 fungal. Twenty-two bacterial infections produced positive cultures by direct plating, and all produced the same organism with Amies medium after 4 and 24 hours, respectively. For 43 fungal infections identified by KOH stain, 29 (67%) yielded a positive result after 4 hours in Amies transport medium and 27 (63%) after 24 hours in Amies medium. A total of three cases (7%) that showed fungal infection on KOH stain but did not yield organisms by direct plating did so after inoculation with Amies transport medium. For all comparisons, there was no difference in recovery rates by means of transport medium compared with direct plating (McNemar exact P >.05). CONCLUSIONS In the clinical setting, Amies transport medium may be a useful alternative to direct inoculation onto blood agar for the laboratory evaluation of infectious keratitis.

A new type of swab (Cellswab; Cellomeda, Turku, Finland), utilizing a highly absorbent cellulose viscose sponge material, was compared to some traditional swabs. The survival of 14 aerobic and 10 anaerobic and microaerophilic bacterial species in the Cellswab, two commercial swab transport systems (Copan, Brescia, Italy, and Orion Diagnostica, Espoo, Finland), and one Dacron swab (Technical Service Consultants Ltd.[TSC], Heywood, United Kingdom) was evaluated. Bacteria were suspended in broth, into which the swabs were dipped. The Cellswab absorbed 1.3 times more fluid and released 3.5 times more fluid upon plating than the other swabs. Aerobic bacteria were stored in dry tubes, the others in transport medium, at 4 degrees C and room temperature (RT), for up to 14 days. Swab samples were transferred to plates at 0, 1, 2, 4, 7, and 14 days. For 10 strains the Cellswab yielded > or =10% of the original CFU for longer than all the other swabs. In the clinical study, the ability of the Cellswab to detect beta-hemolytic streptococci from throat samples (n = 995) was compared to that of the TSC Dacron swab. The swabs performed equally, both when their samples were transferred to plates immediately and after storage for 1 day at 4 degrees C or RT. The changes in normal microbiota after storage were also similar. The Cellswab was found to perform at least as well as ordinary swabs. It was better at storing fastidious strains, and at keeping bacteria viable for long storage times; it might well be a useful replacement or complement to ordinary swabs.

1. The transport of negatively charged drugs, xenobiotics, and metabolites by epithelial tissues, particularly the kidney, plays critical roles in controlling their distribution, concentration, and retention in the body. Thus, organic anion transporters (OATs) impact both their therapeutic efficacy and potential toxicity. 2. This review summarizes current knowledge of the properties and functional roles of the cloned OATs, the relationships between transporter structure and function, and those factors that determine the efficacy of transport. Such factors include plasma protein binding of substrates, genetic polymorphisms among the transporters, and regulation of transporter expression. 3. Clearly, much progress has been made in the decade since the first OAT was cloned. However, unresolved questions remain. Several of these issues--drug-drug interactions, functional characterization of newly cloned OATs, tissue differences in expression and function, and details of the nature and consequences of transporter regulation at genomic and intracellular sites--are discussed in the concluding Perspectives section.

Doxepin is a tricyclic antidepressant that is widely prescribed for the treatment of mild depression. In this study, hair samples collected from a patient receiving 25 mg of doxepin daily were analyzed. Doxepin was administered to the patient for 4 months (June 15 to October 15, 1996). Five hair samples were collected: 1 and 3 months after doxepin therapy began and 1, 3, and 5 months after drug therapy ended. Solid-phase extraction was employed to isolate doxepin and its major metabolite desmethyldoxepin from the hair matrix, and gas chromatography-mass spectrometry (GC-MS) was used for quantitation of both drugs. Six-point standard curves (0.25-20 ng/mg) were prepared for both compounds with an internal standard (doxepin-d3). The standard curves for doxepin and desmethyldoxepin were linear over the range reported and had correlation coefficients of 0.984 and 0.985, respectively. The limit of quantitation for both analytes was 0.25 ng/mg of hair. In addition, the replicate analysis of control hair preparations was performed at two levels (2 ng/mg and 15 ng/mg) to determine intra- and interday variability. Doxepin and desmethyldoxepin were not detected in the patient's sample collected 1 month after doxepin therapy began. The samples collected 3 months after doxepin therapy began and 5 months after drug therapy was terminated had detectable amounts of doxepin and desmethyldoxepin. The highest concentrations of doxepin (mean, 0.59 ng/mg) and desmethyldoxepin (mean, 0.40 ng/mg) were found 5 months after doxepin therapy began, which was also 1 month after the patient had stopped using the drug. Five months after doxepin therapy was terminated, the drug and its metabolite were still present in the patient's hair. The concentration of doxepin in hair was always significantly higher than the concentration of desmethyldoxepin.

A mutant of Cochliobolus heterostrophus lacking the outer layer of extracellular matrix around its germ tubes and hyphae was obtained by mutagenizing protoplasts. The mutant not only lacks the outer matrix, but also produces much smaller lesions on corn leaves than nonmutant strains; the area of mutant lesions averages 0.6 mm2 compared with 5.8 mm2 for nonmutant lesions. Genetic analysis demonstrated that the failure to produce the outer matrix cosegregates with the reduced lesion size, indicating that the two traits are controlled by the same locus, designated Ecm1 (Extracellular Matrix Deficient). The mutant retains normal growth on media and normal abilities to germinate, form appressoria, and penetrate corn leaves. This indicates that the outer matrix is not necessary for infection prior to entrance of the fungus into the leaf. It also indicates that the pathogenicity defect in this mutant is manifested after penetration. To facilitate future tests of whether the pathogenicity defect is caused by the lack of the outer matrix, Ecm1 was mapped. Seven markers linked to Ecm1 were found by analysis of amplified fragment length polymorphisms. Ecm1 maps to chromosome 4; the closest markers to Ecm1 are 5 cM distant, which is estimated to represent about 115 kb.

This retrospective cross-sectional study explored the associations of personality characteristics with parenting problems among 25 couples, one or both members of which were identified as alcoholics by virtue of their voluntary past completion of a residential program for alcoholics. Most of them (90%) scored lower, indicating their more problematic parental attitudes and behaviors, on all four scales of the Adult-Adolescent Parenting Inventory (AAPI: inappropriate parental expectations of children, lack empathy for children's needs, value physical punishment, and parent-child role reversal) than average "normal" nonalcoholic, nonabusive adults. Such parenting problems were found to be very highly associated with clients' personality characteristics. For example, schizoid, schizotypal, histrionic, and passive aggressive characteristics (DSM-III-R-based) along with a few other personal characteristics of the couples, accounted for nearly all (90.2%, R2 =.902) of their propensity to reverse roles with their children. Findings also suggested that the identified parenting problems among alcoholic couples are amenable to programmatic intervention: the longer couples had participated in aftercare programs offered by the treatment facility the more appropriate and empathetic was their parenting.

The duration of behavioral impairment after marijuana smoking remains a matter of some debate. Alcohol and marijuana are frequently used together, but there has been little study of the effects of this drug combination on mood and behavior the day after use. The present study was designed to address these issues. Fourteen male and female subjects were each studied under four conditions: alcohol alone, marijuana alone, alcohol and marijuana in combination, and no active treatment. Mood and performance assessments were made during acute intoxication and twice the following day (morning and mid-afternoon). Acutely, each drug alone produced moderate levels of subjective intoxication and some degree of behavioral impairment. The drug combination produced the greatest level of impairment on most tasks and "strong" overall subjective ratings. There were few significant interactions between the two drugs, indicating that their effects tended to be additive. Only weak evidence was obtained for subjective or behavioral effects the day after active drug treatments, although consistent time-of-day effects (morning versus afternoon) were observed on several subjective and behavioral measures. In sum, this study provided little evidence that moderate doses of alcohol and marijuana, consumed either alone or in combination, produce behavioral or subjective impairment the following day.

Alcohol and marijuana are frequently used together, yet there has been little study of how the presence of one drug might affect consumption of the other. The present study examined the effects of alcohol pretreatments on marijuana self-administration in a group of 15 males and 5 females who were users of both drugs. During evening sessions in a recreational setting, pairs of subjects consumed drinks containing 0.0, 0.3 or 0.6 g/kg alcohol 30 min before a 60-min period of ad libitum marijuana smoking. Marijuana self-administration was assessed in several ways: by measuring the number of cigarettes smoked, the increase in expired air carbon monoxide resulting from marijuana smoke inhalation, and the increase in heart rate due to THC absorption. None of these variables was significantly affected by the alcohol pretreatments, although substantial individual differences were observed. These results indicate that low to moderate doses of alcohol do not systematically influence marijuana self-administration.

Three commercially available systems, the 4-h Minitek Enterobacteriaceae III, the Haemophilus Trio-Tube, and the Micro-ID, were evaluated for their capacities to identify and biotype 308 respiratory isolates of Haemophilus spp. When compared with aminolevulinic acid test results, the definitive identification method used in this study, these systems demonstrated no significant differences in their capacities to differentiate Haemophilus influenzae from Haemophilus parainfluenzae. They were in agreement with the standard method of species identification approximately 50% of the time. When sucrose was added to the Minitek and Trio-Tube configurations, the efficiency rate of species identification increased to more than 95%. The Micro-ID could not be modified to incorporate this additional biochemical parameter. The performance of the sucrose-supplemented Minitek and Trio-Tube systems, compared to the combined results of Micro-ID and aminolevulinic acid, produced correlations of 94 and 90%, respectively. Rapid and accurate methodologies are available for combined species identification and biotyping of Haemophilus spp.

Microscopy and leukocyte esterase activity, both employed as screening techniques for urine cultures, were evaluated with respect to two distinct populations, male and female. When 424 urine specimens from males were examined, 95% of the Gram-stained smears and 91% of the leukocyte esterase tests correctly correlated with culture results, indicating significant bacteriuria. There were no significant differences between the Gram stain and leukocyte esterase activity in predicting a negative culture: 99% and 98%, respectively. Neither microscopy nor esterase activity proved as sensitive or as efficient in predicting a negative culture with the female population.

A highly efficient method is described for producing at room temperature anoxic solutions of 50 ml or less in test tubes or serum vials by combining negative pressure with strong vortexing so that the liquid-surface, gas exchange area is increased by orders of magnitude. Liquid media suitable for the cultivation of methanogens may be rendered anoxic after three short vacuum-vortex steps.

The spacing between optical amplifiers in a long-haul soliton system may be increased to 100 km by using only passive quantum-well saturable absorbers and narrow-band filters for soliton control. After transmission over 9000 km at 10 Gbits/s, the effects of soliton-soliton interaction and Gordon-Haus jitter in the proposed systemyield bit error rates of better than 10(-9).

The exclusion of plate counts outside of an acceptable range can bias the estimation of concentration. If these plates are too numerous to count (TNTC), then their microbes may be inhibited. If inhibition of the microbes on a plate is suspected, then its count may be best treated as a lower bound. When these lower bounds replace counts for some plates, the estimate and confidence interval must be calculated accordingly. Also, a measure of unusualness for plate counts is discussed.

A technic is described whereby poliomyelitis virus may be sedimented by centrifugation for four hours at 18,000 r.p.m. Virus has been recovered quantitatively in the sediment from a 20 per cent., but not a 1 per cent., saline suspension. The addition of 10 per cent. normal serum results in quantitative recovery from a 1 per cent. suspension. Virus has been recovered by intracerebral inoculation from two of four human stools tested by the method described.

In total, 172 isolates of Enterobacteriaceae, Acinetobacter spp., Pseudomonas aeruginosa and Stenotrophomonas maltophilia were tested for susceptibility to colistin by agar dilution, Etest and the Vitek 2 system. Isolates with a colistin MIC < or =2 mg/L were considered to be susceptible. Fifty-four (31%) Gram-negative isolates were resistant to colistin. Categorical agreement between agar dilution and Etest was 87%, and between agar dilution and Vitek 2 was 82%. Based on the data obtained, the Vitek 2 system was unreliable for detecting colistin resistance, and results obtained by Etest may require confirmation by a standard MIC susceptibility testing method.

Trismus is a more common symptom in NPC patients with young age and an indicator of advanced primary tumour. Overall response rate after treatment was 88%. Trismus recovered in majority of patients at the end of treatment and patients with complete recovery of trismus may have a better survival.

Cervical and mediastinal cystic lymphangioma are rare and benign tumours. They looked like a lateral and cervical mass. Medical imaging allows to suspect the diagnosis but histology only may to assert it. The best therapy is surgery. Sclerosis by chemical drugs is only an alternative if surgery is not possible. A recovery is completely obtained if the whole tumour is removed.

The Bactec MGIT 960 system (Becton Dickinson, USA), designed for the culture of mycobacteria, was compared with the Bactec 460 instrument and culture on two solid egg-based media using a total of 1024 clinical specimens. Mycobacteria could be identified from 99 (9.7%) specimens, 89 (90%) of which were identified by the Bactec 960 system, 90 (91%) by the Bactec 460 system, and 82 (83%) by culture on the two egg-based media. The Bactec 960 cultures became positive an average of 16.7 days after specimen collection, the Bactec 460 cultures 14.9 days after collection, and the cultures on egg-based media 26.2 days after collection. The Bactec 960 is a compact and highly automated nonradiometric system that may replace the Bactec 460 system.

A recent literature study has shown that after 3 days of use of a transdermic patch of fentanyl (Durogésic(R)), there was such an amount of drug remaining that may make its excessive use possible. First of all, we have developed a fentanyl assay by a spectrophotometric method and then, the remaining amount of fentanyl was determined in 29 used patches. This study has shown that a mean of 22% of fentanyl remained in the used patches, each original content astounded. Then, we have determined an easy and feasible method in a chemist's shop allowing inactivation of fentanyl in the used patches and so preventing its diversion.

The central nervous system may be affected by AIDS in the form of infections or neoplasia and by HIV itself. The spine and spinal cord are involved more commonly than documented. MR imaging is the most sensitive imaging technique for the detection of AIDS-related abnormalities in the spine and spinal cord. The early detection by spinal imaging will allow the prompt initiation of the appropriate therapy.