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We show that a higher incidence of campylobacteriosis is found in young children (age, <5 years) living in rural, compared with urban, areas. Association of this difference with particular animal sources was evaluated using multilocus sequence typing. This evaluation was achieved by comparing Campylobacter isolates originating from these children, retail poultry, and a range of animal sources by use of source attribution and phylogenetic analysis methods. The results indicate that chicken is a major source of infection in young urban children, although not in their rural counterparts, for which ruminant and other avian sources are more important.

Pairs of individuals tested at the 13 CODIS core STR loci to determine sibship were used as a source of familial data that was seeded into a larger data set of 12,000 plus DNA profiles simulating a CODIS-like offender database. To determine whether known sibs could be found in the larger database two methods were used: degree of allele sharing and a kinship matching approach. The allele sharing method detected 62 of 109 of the known sib pairs (57%) while kinship matching detected 90 of the sib pairs (83%). Although kinship matching was the more efficient method of the two, the number of false positives generated prior to finding a true match was inversely related to the likelihood of sibship suggesting that many true siblings would not be easily found in a large forensic database via familial searching techniques.

We have examined 389 father/son sample pairs from U.S. Caucasians, African Americans, Hispanics and Asians using the 17 Y-STR loci in the Yfilertrade mark kit and observed a total of 24 differences between father and son. Thirteen mutations resulted in the gain of a repeat in the son and 11 resulted in a loss of a repeat. All samples resulted in single repeat mutations except one sample which contained a two repeat loss at Y-GATA-H4. Furthermore, two different sample pairs were found to have two mutations. An African American sample pair had a mutation at DYS458 and a second at DYS635 and an Asian sample pair had mutations at DYS439 and Y-GATA-H4.

Leucine is recognized as a nutrient signal; however, the long-term in vivo consequences of leucine signaling and the role of branched-chain amino acid (BCAA) metabolism in this signaling remain unclear. To investigate these questions, we disrupted the BCATm gene, which encodes the enzyme catalyzing the first step in peripheral BCAA metabolism. BCATm(-/-) mice exhibited elevated plasma BCAAs and decreased adiposity and body weight, despite eating more food, along with increased energy expenditure, remarkable improvements in glucose and insulin tolerance, and protection from diet-induced obesity. The increased energy expenditure did not seem to be due to altered locomotor activity, uncoupling proteins, sympathetic activity, or thyroid hormones but was strongly associated with food consumption and an active futile cycle of increased protein degradation and synthesis. These observations suggest that elevated BCAAs and/or loss of BCAA catabolism in peripheral tissues play an important role in regulating insulin sensitivity and energy expenditure.

Likelihood ratios (LRs) were calculated for a cohort of 60 pairs of true half-sibs and compared with LR values calculated for unrelated, paired children. STR results for the half-sib group were obtained from 60 archived cases involving a true mother, two children, and an alleged father subjected to typing with a multiplex STR kit (Identifiler multiplex, Applied Biosystems) and in which the alleged father was excluded as the father of only one of the two children (half-sib pairs). The distribution of LR values among true half-sibs was compared to those produced from paired, unrelated children selected in two ways: One method for producing unrelated pairs was to randomly select Identifiler profiles from children in 120 distinct paternity cases and group them into 60 ethnically matched pairs (random pairs). In a second approach, the children in the true half-sib group were shuffled and ultimately paired with someone from a different case. A total of 49 ethnically matched, unrelated pairs were created (shuffled pairs). In the shuffled pairs group, comparisons were thus based on a constant set of phenotypes. LRs comparing the probability of half-sibship versus being unrelated were produced for all groups with standard methods. Among pairs of known half-sibs, LRs ranged from a low of 0.1 to a maximum of 3763. Among random and shuffled pairs, LRs ranged from a low of 0.0001 to 12 for shuffled pairs or 42 for random pairs. LRs of greater than 2 were produced in 8 instances among random pairs and in 4 instances among the shuffled pairs. Overall, results suggest that half-sib indices of 30 or greater are fairly characteristic of individuals who are related as half-sibs. In contrast, half-sib indices of 0.1 or less are fairly characteristic of unrelated individuals who claim to be half-sibs. LRs falling between 0.1 and 10.0 are uninformative, as this region represents the overlap in the LR distributions produced from the true and false half-sib groups when the Identifiler multiplex kit is used for testing.

The three main pathways of Escherichia coli O157 infection are foodborne, environmental (including direct contact with animals and their faeces and contaminated water supplies) or person to person contact. The disease is often nicknamed the 'burger bug' but it appears that environmental risk factors may be more important. In this study we use four techniques (outbreak analysis, case-control studies, disease mapping and quantitative microbial risk assessment (QMRA)) to determine whether burgers or environmental pathways present the greater risk in Scotland. Analysis of E. coli O157 outbreaks in Scotland from 1994 to 2003 associated with either meat or dairy foods, or with environmental transmission shows that approximately 40% [M1] of these outbreaks were foodborne, 54% were environmental and 6% involved both transmission routes. However, the largest outbreaks tend to be foodborne accounting for 83% of outbreak cases. Case-control studies indicate strong risk associations with environmental exposure in Scotland, the UK as a whole and the USA, but burgers appear to be more of a risk in the USA. Canadian, Scottish and Swedish disease mapping studies found positive association with indicators of cattle density. In Grampian (North-East Scotland) we found that there was a positive association with cattle and sheep density (divided by human population density) as well as percentage of population on private water supplies. We found 63% of cases in rural postcodes compared with 37% urban after correcting for population differences suggesting that at least 26% of cases may be classified as environmental. QMRA showed that on average, the risk was 100 times greater when visiting a pasture than eating a burger in Grampian. However, it is difficult to determine which pathway actually causes most illnesses as it is unknown how many burgers are consumed daily and what is the frequency of human visits to pasture. The implementation of hygienic food processing post-1996 Central Scotland outbreak and the preference for 'well done' burgers may account for this food being a relatively low risk thus making the 'burger bug' term less appropriate in the UK.

Objective: To compare the effect of a combination of 20% citric acid solution and photo-activated disinfection with the use of 20% citric acid and 2.25% sodium hypochlorite solutions on bacterial load on the dentine walls in prepared canals in vivo., Subjects and Methods: Sixty-four randomly selected cases were evaluated and allocated to one of two groups. In Group 1, after gaining access to the root canal, bacterial load on the canal walls was sampled using endodontic files. A further sample was taken after apex location and initial widening of the canal had been completed and the photo-activated disinfection process carried out. A final sample was taken after completion of the canal preparation using citric acid and sodium hypochlorite solutions. In Group 2, the initial sample was taken as described previously. A second sample was taken after conventional preparation using 20% citric acid and sodium hypochlorite solutions as co-irrigants. A final sample was then taken after a subsequent PAD treatment. All samples were cultured for facultative anaerobic bacteria., Results: Of the canals treated in Group 1 only two of the 23 canals infected showed culturable bacteria after the use of citric acid and photo-activated disinfection. Of these two canals, one was free of culturable bacteria on completion of conventional treatment but the other still contained culturable bacteria. In Group 2, four canals of the 23 infected initially, remained contaminated after conventional treatment. After subsequent photo-activated disinfection three of these four canals were free of culturable bacteria., Conclusion: Results indicate that the use of a chelating agent acting as a cleaner and disrupter of the biofilm and photo-activated disinfection to kill bacteria is an effective alternative to the use of hypochlorite as a root canal cleaning system.

Objective: To determine the microbiological effect of photoactivated disinfection (PAD) as an adjunct to normal root canal disinfection in vivo., Design: A randomised trial carried out in general dental practice., Subjects and Methods: Patients presenting with symptoms of irreversible pulpitis or periradicular periodontitis requiring endodontic therapy were selected at random. A microbiological sample of the canal was taken on accessing the canal, after conventional endodontic therapy, and finally after the PAD process (photosensitiser and light) had been carried out on the prepared canal. All three samples from each canal were plated within 30 minutes of sampling and cultured anaerobically for five days. Growth of viable bacteria was recorded for each sample to determine bacterial load., Results: Thirty of the 32 canals were included in the results. Cultures from the remaining two did not reach the laboratory within the target time during which viability was sustained. Of the remaining 30, 10 canals were negative to culture. These were either one of the canals in multi rooted teeth where the others were infected or where a pre-treatment with a poly-antibiotic paste had been applied to hyperaemic vital tissue. Sixteen of the remainder were negative to culture after conventional endodontic therapy. Three of the four which had remained infected cultured negative after the PAD process. In the one canal where culturable bacteria were still present, a review of the light delivery system showed a fracture in the fibre reducing the effective light output by 90%., Conclusions: The PAD system offers a means of destroying bacteria remaining after using conventional irrigants in endodontic therapy.

Background: Campylobacter infection is a major cause of bacterial gastrointestinal disease. Exposure to Campylobacter is known to produce an immune response in humans that can prevent future symptomatic infections. Further, studies of the general population have shown that seroprevalence to Campylobacter increases with age., Methods: A large collection of serotyped Campylobacter isolates, obtained from human clinical faecal samples, were analysed by comparing the ratio of uncommon to common serotypes by different age groups, using chi2 tests., Results: We have identified that older age groups, as well as having generally lower incidence, are significantly less likely to be infected by the more common serotypes., Conclusion: These results are indicative of acquired immunity, however, further studies are needed to rule out the confounding effects of the variations in exposure pathways experienced by different age groups.

Fifty known siblings and fifty unrelated pairs were genotyped using the ABI Identifiler STR system and sibship indices computed for each pair. Combined sibship indices (CSIs) for the known siblings ranged from less than 10 to greater than 1 billion. CSIs for the unrelated pairs ranged from 4.5 x 10(-8) to 0.12. In the known sibling group the percentage of loci where both alleles matched was approximately 40%, while the percentage of loci where neither matched was approximately 10%. In the non-sibling group, the percentage of loci where both alleles matched was approximately 6%, while the percentage of loci where neither matched was approximately 45%. Interestingly, the percentage of loci where a single allele matched was the same in both the known siblings and unrelated pairs, approximately 50%.

Aims: To compare immunomagnetic separation (IMS) protocols (enrichment media and temperature) for the isolation of Escherichia coli serotypes O26 and O111 from four different foods., Methods and Results: Foods (minced beef, cheese, apple juice and pepperoni) spiked with low numbers (<100 g(-1)) of stressed nalidixic mutant E. coli serotypes O26 and O111 were enriched in media based on buffered peptone water (BPW), tryptone soya and EC broths incubated at temperatures of 37 and 42 degrees C to optimize the IMS technique. BPW enrichments gave increased recoveries of both serotypes compared with tryptone soya and EC broths. Elevated temperatures of incubation at 42 degrees C were superior to 37 degrees C., Conclusions: Positive detection of low numbers of stressed target pathogens in all replicate tests was only possible using BPW enrichments. The majority of tests from alternative enrichments resulted in zero or single colonies recovered post-IMS., Significance and Impact of the Study: The optimum IMS protocol would improve isolation rates of E. coli O26 and O111 from foods and lead to increased safety for the consumer. Sub-optimal IMS protocols could lead to foods being incorrectly labelled free from these pathogens.

Objective: To determine the potential for asphalt fume exposure to increase DNA damage, we conducted a cross-sectional study of roofers involved in the application of roofing asphalt., Methods: DNA strand breaks and the ratio of 8-hydroxydeoxyguanosine (8-OHdG) to 2-deoxyguanosine (dG) were measured in peripheral blood leukocytes of roofers. In addition, urinary excretion of 8-OHdG and 8-epi-prostaglandin F2alpha (8-epi-PGF) was also measured. The study population consisted of 26 roofers exposed to roofing asphalt and 15 construction workers not exposed to asphalt during the past 5 years. A subset of asphalt roofers (n = 19) was exposed to coal-tar pitch dust (coal tar) during removal of existing roofs prior to applying hot asphalt. Personal air monitoring was performed for one work-week to measure exposure to total particulates, benzene-soluble fraction of total particulates, and polycyclic aromatic compounds (PACs). Urinary 1-OH-pyrene levels were measured as an internal biomarker of PAC exposure., Results: Full-shift breathing zone measurements for total particulates, benzene-solubles and PACs were significantly higher for coal-tar exposed workers than for roofers not exposed to coal tar. Similarly, urinary 1-OH-pyrene levels were higher in coal-tar exposed roofers than roofers not exposed to coal tar. Total particulates or benzene-soluble fractions were not associated with urinary 1-OH-pyrene, but PAC exposure was highly correlated with urinary 1-OH-pyrene. When stratified by 1-OH-pyrene excretion, DNA strand breaks increased in a dose-dependent manner, and leukocyte 8-OHdG/dG decreased in a dose-dependent manner. Significant changes in DNA damage appeared to be linked to PACs from coal-tar exposure, although asphalt fume alone was associated with a small but significant increase in urinary 1-OH-pyrene and DNA strand breaks., Conclusions: Results are consistent with previous reports that asphalt or coal-tar exposure can cause DNA damage. Urinary 8-epi-PGF remained relatively constant during the week for virtually all subjects, regardless of exposure indicating that neither asphalt nor coal-tar exposure induces an overt oxidative stress. A small, but statistically significant increase in 8OHdG was evident in end-of-week urine samples compared with start-of-week urine samples in roofers exposed to coal-tar. The increase in urinary 8OHdG coupled with the decrease in leukocyte 8-OHdG/dG, suggests that coal-tar exposure induces protective or repair mechanisms that result in reduced levels of steady-state oxidative-DNA damage.

An outbreak of E. coli O157 infection occurred in the Highland Region of Scotland in the summer of 1999. The source of the outbreak was traced to an untreated private water supply. All six cases identified arose in visitors to the area, and most had very limited exposure to the contaminated water. Permanent residents on the same supply were unaffected. The E. coli O157 isolates from the water, sheep faeces collected from around the source and the human stool samples were indistinguishable using pulsed field gel electrophoresis. Previously reported outbreaks of E. coli O157 linked to potable water supplies have resulted from structural or treatment failures, which allowed faecal contamination of source water. Here, contamination of the water supply and subsequent human infection was due to the use of an untreated, unprotected private water source in a rural area where animals grazed freely.

Hepatitis B virus (HBV) infection was transmitted by a locum cardiothoracic surgeon to two patients during coronary artery bypass surgery. Both patients presented 12 weeks after surgery and developed serious clinical illness. The surgeon was known to be hepatitis B surface antigen (HBsAg) positive, hepatitis B e antigen (HBeAg) negative, and to have antibodies to HBeAg (anti-HBe). Sequences of regions of the HBV surface and core genes from the patients and surgeon were indistinguishable. An exercise was undertaken to notify all patients on whom the surgeon had operated while employed at the hospital where the transmissions occurred. One hundred and twenty-three out of 126 patients were tested. No evidence of transmission to any other patient was found. Revised recommendations by the UK Health Departments as to which health care workers should be permitted to perform exposure prone procedures have recently been published.

Chlorpyrifos is a moderately toxic organophosphate pesticide. Houses and lawns in the United States receive a total of approximately 20 million annual chlorpyrifos treatments, and 82% of U.S. adults have detectable levels of a chlorpyrifos metabolite (3,5, 6-trichloro-2-pyridinol; TCP) in the urine. The U.S. Environmental Protection Agency has estimated that there are 5,000 yearly reported cases of accidental chlorpyrifos poisoning, and approximately one-fourth of these cases exhibit symptoms. Organophosphates affect the nervous system, but there are few epidemiologic data on chlorpyrifos neurotoxicity. We studied neurologic function in 191 current and former termiticide applicators who had an average of 2.4 years applying chlorpyrifos and 2.5 years applying other pesticides, and we compared them to 189 nonexposed controls. The average urinary TCP level for 65 recently exposed applicators was 629.5 microg/L, as compared to 4.5 microg/L for the general U.S. population. The exposed group did not differ significantly from the nonexposed group for any test in the clinical examination. Few significant differences were found in nerve conduction velocity, arm/hand tremor, vibrotactile sensitivity, vision, smell, visual/motor skills, or neurobehavioral skills. The exposed group did not perform as well as the nonexposed group in pegboard turning tests and some postural sway tests. The exposed subjects also reported significantly more symptoms, including memory problems, emotional states, fatigue, and loss of muscle strength; our more quantitative tests may not have been adequate to detect these symptoms. Eight men who reported past chlorpyrifos poisoning had a pattern of low performance on a number of tests, which is consistent with prior reports of chronic effects of organophosphate poisoning. Overall, the lack of exposure effects on the clinical examination was reassuring. The findings for self-reported symptoms raise some concern, as does the finding of low performance for those reporting prior poisoning. Although this was a relatively large study based on a well-defined target population, the workers we studied may not be representative of all exposed workers, and caution should be exercised in generalizing our results.

Occupational nitrosamine exposures from a rubber vehicle seal (VS) curing operation were compared with the peripheral blood lymphocyte concentrations of two nitrosamine-related DNA adducts, N(7)-methylguanine (N(7)mdG) and O(6)-methylguanine (O(6)mdG), and with the activity of the enzyme that repairs O(6)mdG adducts, O(6)-alkylguanine-DNA alkyltransferase (AGT). The occupational personal breathing zone (PBZ) nitrosamine exposures ranged from 0.4 to 9.3 microg/m(3) in the VS area, from 0.1-2 microg/m(3) in an area remote from the VS and were not detected at a nearby rubber plant. Workers from all three of these locations had detectable concentrations of N(7)mdG adducts, ranging from 0.1 to 133.2 adducts/10(7) deoxyguanosine nucleosides. Although N(7)mdG concentrations were elevated for those who worked in the VS area (median 3.60 compared with 1.44), the difference was not statistically significant after controlling for confounding factors. The O(6)mdG adduct concentrations were much lower than those of N(7)mdG, ranging from non-detectable to 12.7 O(6)mdG adducts/10(7) deoxyguanosine nucleosides and many of the participants (40/78 successfully analyzed) did not have detectable amounts of these adducts (limit of detection 0.03 O(6)mdG adducts/10(7) deoxyguanosine nucleosides). Analysis of the ordinal exposure categories (high, medium/high, medium/low, low and no exposure) yielded a statistically significant association with having detectable O(6)mdG adducts (Kendall's taub = -0.253, asymptotic SE = 0.096). There was no significant association between AGT activity and nitrosamine exposure or exposure category (P > 0.30). Although no association was found between PBZ exposure and either the N(7)mdG adduct concentrations or AGT activity, the significant positive association between working in and near the VS department and the presence of O(6)mdG adducts, which have mutagenic potential, provides evidence to link nitrosamine exposure one step closer to human cancer by demonstrating an association between external nitrosamine exposures and cancer-related biological effects.

Four patients with Salmonella enteritidis infection are reported. All were body builders who regularly consumed substantial quantities of raw eggs. They presented with a severe febrile illness and diarrhoea--presumably reflecting a large bacterial inoculum. Advice regarding the potential hazards of raw egg ingestion has been repeatedly issued by the Department of Health--but this report highlights the fact that this practice continues in spite of this. The epidemiology of S. enteritidis infection in relation to raw egg ingestion is discussed.

The genotoxic potential of two occupationally significant chemicals, 4,4'-methylene-bis-2-chloroaniline (MOCA) and 2-phenyl-1,4-benzoquinone (PBQ), was explored by monitoring the induction of mutations at the HPRT locus of AHH-1 human lymphoblastoid cells. Exposure of AHH-1 cells to the putative carcinogenic metabolite of MOCA, N-OH-MOCA, induced a 6-fold increase in mutant frequency and resulted in base pair substitutions primarily at A:T base pairs. In contrast, exposure to PBQ did not result in an increased mutant frequency although this compound was significantly more cytotoxic than N-OH-MOCA at equimolar doses. The induction of mutations at A:T sites by N-OH-MOCA is consistent with the type of DNA damage known to be produced by MOCA and provides a specific marker of genotoxic damage for exposed populations.

The probable human carcinogen 4,4'-methylene-bis(2-chloroaniline) (MOCA) was utilized to develop biomarkers of exposure to occupational carcinogens. The 32P postlabeling assay, utilizing the nuclease P1 enhancement procedure, was used to evaluate MOCA-DNA adduct formation in target tissues. Male Sprague-Dawley rats were treated with different dosing regimens of MOCA, and DNA was isolated from the liver. Additionally, a human uroepithelial cell (HUC) line was treated with N-hydroxy-MOCA for 24 hr, cells were harvested, and DNA was isolated. DNA was analyzed for MOCA-DNA adduct formation by the 32P postlabeling assay. Five MOCA adducts were detected in rat liver DNA. Adduct A, which corresponded to N-(deoxyadenosin-8-yl)-4-amino-3-chlorobenzyl alcohol, was the major adduct in rat liver DNA appearing in all treatment groups. Levels of adduct A were higher when MOCA was administered by ip injection versus oral gavage. Phenobarbital pretreatment increased the amount of adduct A approximately 12-fold. The pathway leading to the formation of adduct A in DNA from HUC appeared to be saturated at the concentrations used: 2.5, 5, and 10 microM. However, an additional adduct (E) was observed at the 10 microM treatment level only. A major DNA adduct was detected in the target tissue of rats and target human cells for MOCA-induced carcinogenesis, thus making it useful as a biomarker of exposure. Other DNA adducts were also observed with the different doses and routes of exposure investigated.

We report three examples of community-acquired toxigenic Clostridium difficile diarrhoea in elderly patients who had neither received antimicrobial therapy nor been institutionalised. These cases stimulated interest in the non-antimicrobial changes which might predispose the host to C. difficile-related disease and raised the spectre of bowel ischaemia as a possible aetiological factor.

One hundred and sixty-five consecutive patients ( > 2 years of age) with acute group A streptococcal (GAS) pharyngitis randomly received co-amoxyclav (79 patients) or phenoxymethyl penicillin (86 patients). beta-Lactamase activity in saliva was determined for each patient. At follow up after seven days, tonsillar cultures from seven patients (9.6%) in the penicillin V group grew group A streptococcus; three of these patients had tonsillitis clinically. In the co-amoxiclav group these figures were three (3.8%) and two respectively (P > 0.05). Within the 12 month follow up period, there were four clinical recurrences (6.1%) in the penicillin V group and seven (9.3%) in the co-amoxiclav group (P > 0.1). beta-Lactamase activity in the saliva was demonstrated in 29 patients (19.2%). Fourteen (74%) of 19 bacteriological failures or clinical recurrences had beta-lactamase activity, versus 15 (12%) of 129 successfully treated patients (P < 0.001). There is no evidence that oral co-amoxiclav is better than oral penicillin V for the first treatment of acute GAS pharyngitis, but bacteriological failure and clinical recurrence are strongly associated with the presence of beta-lactamase activity in commensal flora.

Objective: To study the effect of prophylactic antibiotics on the incidence of bacteraemia following hysteroscopic surgery., Design: Prospective randomized study., Setting: Aberdeen Royal Infirmary., Subjects: One hundred and sixteen women about to undergo either endometrial laser ablation (ELA) or transcervical resection of the endometrium (TCRE)., Intervention: Fifty-five women were randomised to receive 1.2 g of Augmentin (co-amoxiclav) i.v. at induction of anaesthesia. Sixty-one women received no antibiotic prophylaxis. Blood cultures were obtained at the end of the surgical procedure., Results: Incidence of bacteraemia in the non-antibiotic group (16%) was significantly higher than that in the antibiotic group (2%) (95% confidence interval for difference from 5% to 25%). The majority of organisms were of dubious clinical significance and contamination could not be excluded in 7 cases out of 10., Conclusion: There is no convincing evidence that antibiotics are of value in this clinical setting.

Verotoxin-producing strains of Escherichia coli (VTEC), in particular serotype O157:H7, are now recognised as the major cause of haemorrhagic colitis and the haemolytic uraemic syndrome (HUS) in the U.K. and in North America, and increasingly so in other countries. Over a 3-year period (1989-1991), 16 cases of E. coli 0157 infection occurred in one town (Peterhead) in north-east Grampian. Four patients required admission to hospital, of whom three developed HUS. The bovine source of VTEC infection has now been clearly established with foodborne, waterborne, person-to-person and zoonotic transmission described. Despite extensive local enquiries, the source(s) of infection of the 16 cases in Peterhead was not established. Much still needs to be learned about the epidemiology, risk factors and long-term clinical sequelae of VTEC infection and HUS. Close collaboration between the medical and veterinary professions is of paramount importance in order to provide better understanding of the prevalence of E. coli O157 infection in cattle and the route(s) of transmission to humans.

Background: The efficacy of an antibiotic is related to its concentration at the site of infection. Previous studies of the concentrations of amoxycillin and clavulanic acid (co-amoxiclav) in respiratory secretions or whole lung tissue have suffered from methodological problems. The concentration of amoxycillin and clavulanic acid was determined in bronchial mucosal biopsy samples obtained at bronchoscopy following five different dosing regimens., Methods: Bronchial biopsy and serum samples were obtained from 50 patients undergoing diagnostic bronchoscopy. Ten patients each received 375 mg, 625 mg, 750 mg, and 3.25 g oral, and 1.2 g intravenous co-amoxiclav 1-3 hours before bronchoscopy. The concentrations of clavulanic acid and amoxycillin were determined by high performance liquid chromatography using a microbore column, solid phase extraction, and preconcentration to improve sensitivity tenfold over previous methods., Results: Concentrations of both clavulanic acid and amoxycillin in bronchial mucosa were dose related and were well above the MIC90 of co-amoxiclav for the common bacterial respiratory pathogens including Haemophilus influenzae, Micrococcus catarrhalis and Streptococcus pneumoniae for all dosing regimens. Mean mucosal levels were 200% and 118% of the corresponding serum levels for amoxycillin and clavulanic acid respectively., Conclusions: Amoxycillin and clavulanic acid are concentrated in bronchial mucosa and, even at the lowest dose of 375 mg orally, are likely to produce tissue levels in the lung sufficient to inhibit all the common community acquired respiratory pathogens.

A total of 900 consecutive admissions to the Regional Infection Unit at the City Hospital Aberdeen in 1991 have been analysed and the results compared with a similar study during 1980 and 1981. The annual number of admissions increased from 605 to 900, of which 72% in 1991 had proven infections compared with 60% a decade earlier. More patients were admitted with gastroenteritis, tonsillitis and soft tissue infection in 1991 and fewer with non-infectious jaundice. HIV-related conditions contributed 4% of the admissions and 29% of the mortality. Brucellosis disappeared as a reason for requesting hospital admission in North East Scotland.

To assess the contribution of reactive oxygen species (ROS) to metal-induced mutagenesis, we have determined the spectrum of mutations in the lacZ alpha gene after exposure of M13mp2 DNA to Fe2+, Cu2+, and Ni2+. With iron and copper ions, mutations are clustered and are predominantly single-base substitutions. Fe, Cu, and phorbol ester-stimulated neutrophils also produced tandem double CC-->TT mutations. This mutation may provide a marker for the role of oxidative damage in carcinogenesis. Mutagenesis by Ni2+ required the complexing of the metal to a tripeptide and the addition of H2O2. To assess the contribution of ROS in mammalian cells, we determined the spectrum of mutations produced when purified DNA polymerases-alpha and -beta synthesized DNA using a template that had been damaged by ROS. The mutation spectra produced by the two polymerases indicates that these enzymes substitute different nucleotides opposite the same lesions.

In this review we will summarize recent data on reactive oxygen species-induced mutagenesis and consider its relationship to tumorigenesis in humans. With the use of a single-stranded DNA template it has been possible to correlate oxygen radical-induced chemical alterations at specific nucleotides with the types of mutations that occur when these altered bases are copied by DNA polymerases. This has allowed us to identify the types of mutations that occur secondary to a variety of oxidative stresses and study several of the mechanisms by which they arise. The most frequent mutations that result from reactive oxygen species-induced damage to DNA in bacteria are C to T transitions. These mutations, however, are not pathoneumonic for mutagenesis by oxygen-free radicals since they result from DNA damage caused by other genotoxic agents as well as by DNA polymerase errors. One type of mutation, a tandem CC to TT double substitution, has been shown to be induced by reactive oxygen species generated by a variety of systems and may be diagnostic for such damage. In studies with mammalian DNA polymerases, DNA damaged by reactive oxygen species yields mutations different from those observed in Escherichia coli. This diversity of mutagenic changes in these in vitro studies highlights the role of DNA replicating enzymes in specifying the types of mutations produced by reactive oxygen species. In conclusion, we will consider the role of reactive oxygen species in the pathogenesis of three common tumors, carcinoma of the liver, lung, and prostate with consideration on the possible use of antioxidant preventive therapy to slow tumorigenesis sufficiently to prevent clinical presentation of these cancers during the life span of a patient.

Cytosine to thymine transitions are among the most common types of mutations produced by oxygen damage to DNA. One possible mechanism for these transitions is deamination of cytosine to uracil. Using both a forward mutation assay as well as a reversion assay specific for damage to cytosines we show that direct deamination to uracil does not play a significant role in mutagenesis induced by reactive oxygen free radicals. In contrast, lesions sensitive to repair by E. coli endonuclease III play a major role in oxidative mutagenesis as evidenced by the ability of endonuclease III to modulate the extent of mutagenesis that results from exposure of DNA to oxygen free radicals.

We have determined the specificity of mutations produced by nickel(II), a known human carcinogen, in a forward mutation assay and also used a sensitive reversion assay to show that Ni(II), like iron and copper, can produce tandem double CC-->TT mutations, a hallmark of damage to DNA by either UV irradiation or oxygen free radicals. A reduction in mutation frequencies by the addition of oxygen radical scavengers also supports the involvement of reactive oxygen species in DNA damage and mutagenesis by Ni(II). Mutagenesis by Ni(II) is enhanced by the addition of both hydrogen peroxide and a tripeptide glycyl-glycyl-L-histidine. The enhancement of mutagenesis of Ni(II) by the tripeptide indicates that these complexes could serve to localize Ni(II) in nuclei and mediate DNA damage and mutagenesis via the generation of short-lived oxygen free radicals. These data suggest that Ni(II) carcinogenesis may proceed via the generation of active oxygen species and furthermore provide a model for nickel carcinogenesis based on the binding of Ni(II) to nuclear proteins.

Aztreonam blood agar, a new selective medium for Gram positive aerobic bacteria, was evaluated in comparison with conventional media for skin swabs. Aztreonam agar increased the number of isolates of Staphylococcus aureus by 17%. By producing purer growths on primary isolation, it significantly speeded up the identification and sensitivity testing of staphylococci and streptococci. All major Gram positive aerobic pathogens grow on this medium. Aztreonam agar is now an established addition to our culture media. It is used for swabs which are likely to have a mixed Gram positive and Gram negative flora, such as ears, burns, ulcers, and for the sputa of patients with cystic fibrosis.

Oxidative damage to DNA is mutagenic and thus may play a role in carcinogenesis. Because of the large number of different DNA lesions formed by oxidative species, no genetic alteration so far identified is exclusively associated with oxygen damage. Tandem double CC-->TT mutations are known to occur via UV damage to DNA and are thought to be a specific indicator of UV exposure. Using a sensitive reversion assay that can detect both single and double mutations within the same codon of the M13-encoded lacZ alpha gene, we show that treatments that produce reactive oxygen species can also produce tandem double CC-->TT mutations. The frequency at which these mutations occur is less than that for single base mutations by a factor of approximately 30. The induction of these mutations is inhibited by treatment that scavenges hydroxyl radicals. This unique mutation provides a marker of oxygen free radical-induced mutagenesis in cells that are not exposed to UV-irradiation and an indicator for assessing the involvement of oxidative damage to DNA in aging and tumor progression.

Background: Familial cold urticaria is a rare cutaneous and systemic reaction to cold with autosomal dominant inheritance, distinctive clinical features, and unknown pathogenesis. Release of a chymotrypsinlike substance has been postulated. To date, no effective treatment has been reported., Observations: Eight cases from a large family pedigree are described. Three members showed a very favorable response in their cold urticaria, when treated with stanozolol; the response was reproducible. Histologic examination of an early lesion in one case revealed evidence of mast cell degranulation., Conclusions: The biochemical observations are probably secondary epiphenomena. Correction of a deficiency of an inhibitory protein is a possible mechanism of action of stanozolol as in hereditary angioedema.

An important source of endogenous oxygen radicals are phagocytic cells such as neutrophils and macrophages. The human leukemia cell line HL-60 can be induced to differentiate into a neutrophil-like cell population. Among the properties of these differentiated cells is the ability to produce reactive oxygen species when stimulated by tumor promoters. Mutagenesis induced by HL-60-generated free radicals was assessed using the M13mp2 forward mutation assay. Single-stranded M13mp2 DNA was coincubated with phorbol ester-stimulated HL-60 cells, after which mutations were scored by transfecting the DNA into SOS-induced Escherichia coli. The mutation frequency was increased 6-fold above background in DNA incubated with HL-60 cells. The majority of the mutations were single-base substitutions. However, approximately 6% of the mutations were tandem double substitutions that occurred in runs of adjacent cytidines. Overall, the mutations were clustered at apparent "hot spots," many of which were similar to sites seen using iron to generate oxygen radicals. These results suggest that human cells able to produce oxygen radicals in response to tumor promoters might play a significant role in the generation of tumors.

Teicoplanin, a new glycopeptide antibiotic similar to vancomycin, was evaluated in treating 36 hospitalized patients suffering from various Gram-positive infections. The 36 patients received teicoplanin once daily as a mean intravenous injection of 550 mg/day (range 200-800 mg/day). Previous antimicrobial therapy was used in 28% of patients. The mean duration of therapy was 7.5 days (range 3-38 days). The overall clinical success rate was 94%. 24/36 patients (66%) had positive microbiology. Elimination of the pathogens was seen in 75% of all evaluable cases. Four patients with early prosthetic valve endocarditis due to coagulase negative Staphylococcus (3 patients) and Propionibacterium acnes (1 patient) had a favorable clinical and microbiological outcome. No adverse drug reactions were observed. Teicoplanin is safe and effective in the therapy of many different infections caused by Gram-positive bacteria.

We have measured the accuracy of in vitro synthesis by DNA polymerase I (large fragment) during translesion synthesis past an aminofluorene (AF) adduct. These studies were carried out using a site-specifically modified template which contained a single AF adduct. The template was prepared by first modifying the lone guanine in a 17 base long oligonucleotide and extensively purifying and characterizing this product. The modified 17mer was then ligated to a synthetic duplex to produce a 31 nucleotide long template strand containing the AF adduct annealed to a 14mer, such that the 3'-hydroxyl primer terminus was four nucleotides before the modified guanine. Synthesis on this template by DNA polymerase I efficiently bypassed the AF adduct and produced full-length duplex 31mers. T7 DNA polymerase, on the other hand, was unable to utilize the AF-modified template though it was active on an identical unmodified one. The strand synthesized by DNA polymerase I was then separated from the modified strand, annealed to a complementary oligonucleotide, and the resulting heteroduplex cloned into M13. Each of the 49 clones isolated had sequences which indicated that cytidine had been incorporated opposite the AF-modified guanine.

Thirteen cases of group G streptococcal bacteraemia are described. All subjects were hospital in-patients between 1980-1988, and were either middle-aged or elderly. A primary site of infection was identified in 11, of whom nine had soft tissue infection and two had infections derived from the gut/biliary tree. Seven patients had underlying disease, two of whom had malignancy. One patient died of septicaemia. In the majority of patients early institution of treatment led to prompt recovery.

The pharmacokinetics and cardiovascular pharmacodynamics of two oximes were studied in unanesthetized pigs. Effects of 2-[(hydroxyimino)methyl]-1-methylpyridinium chloride (pralidoxime chloride; 2-PAM Cl; 50 mumol/kg) were compared with those of 1,1-methylene bis[4(hydroxyiminomethyl) pyridinium] dichloride (methoxime; MMB-4; 100 mumol/kg). Cardiopulmonary parameters were monitored and plasma concentrations of oximes were determined from arterial blood samples taken at intervals over a period of 5 hr postinjection. Plasma concentrations for both oximes were fitted to standard pharmacokinetic models using the computer program PCNONLIN. Average pharmacokinetic parameters were determined for each oxime. Only mild to moderate physiological side effects were detected following intramuscular administration. 2-PAM Cl was more rapidly absorbed and distributed in the blood than MMB-4. Although the latter had a slight lag time to attain detectable levels in the blood, retention time was longer than that of 2-PAM Cl.

We have analyzed two aspects concerning the relationship between mutations and cancer. Firstly, we consider the possibility that spontaneous mutations result from normal cellular processes. Errors in DNA replication, depurination of DNA, and damage to DNA by oxygen free radicals are important potential sources of endogenous mutations. Mutations produced by DNA polymerases in vitro are clustered, and consist predominantly of single-base substitutions. Analysis of termination sites during copying by DNA polymerase-alpha indicates that certain pause sites are associated with misincorporation of single non-complementary nucleotides, and thus pause sites could be associated with mutagenic hotspots. Depurination results predominantly in transversion mutations resultant from the incorporation of deoxyadenosine opposite abasic sites. Mutations resulting from oxygen free radical damage to DNA are predominantly single-base substitutions: C- > T, G- > C, and G- > T being the most frequent. In addition, there is an unusual mutation that results from oxygen free radical damage to DNA, two adjacent cytidines being substituted by two thymidines. The frequency of mutations observed in tumors appears to be much greater than that predicted from the known rates for spontaneous mutagenesis. Recent observations suggest that many human tumors contain four or more independent mutations. If these multiple mutations are each required for tumor development, then the occurrence of malignancies based on normal mutation rates would be exceedingly rare, and it may be necessary to postulate that tumors result from an increase in the rate of endogenous mutagenesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Twenty-five consecutive patients admitted to hospital with severe soft tissue infection were entered into an open trial designed to evaluate safety and efficacy of the drug combination tazobactam/piperacillin. The dosage regimen was 4 g piperacillin/500 mg tazobactam Q8H. There were twenty cases of uncomplicated cellulitis and five cases with associated abscess formation. These five cases required adjunctive surgical drainage. Mean duration of therapy was 7.6 days. No other antibiotics were administered unless treatment with the study drug failed. There were eight treatment failures, six related to the trial drug. Four patients developed an allergic response to the trial drug, necessitating a change of therapy. Three patients failed to respond; all three had acute cellulitis in association with peripheral vascular disease. Significant bacterial isolates were grown in thirteen patients; Group A streptococci in three, S. aureus in five, other pathogenic streptococci in four and coliforms or Ps. aeruginosa in five. The majority of isolates except the streptococci were resistant to piperacillin but all isolates except one strain of Ps. aeruginosa were susceptible to the combination. The combination is suitable for the treatment of serious soft tissue infection, but increased doses may be appropriate in infection at poorly perfused sites.

A swivel-tethering and jacket system was used in conjunction with vinyl patch electrodes and Holter recorders to obtain continuous ECG recordings in 12 rhesus monkeys on a long-term (12 day) study. Animals were custom-fitted with nylon mesh jackets that were connected to a swivel unit by a flexible, stainless steel tether. Lead wires from the chest electrodes passed through the tether to the electrical swivel apparatus located at the top of the cage. Wires from the upper part of the swivel were attached to a reel-to-reel Holter recorder. This technique was used to obtain 24-h continuous ECG recordings, which were later processed using a computer-assisted Holter analysis system.