An investigation into the association between demographic and morbidity factors, and sleep disturbance.

BACKGROUND: The recognition of sleep disorders is important because in the long term, they are associated with numerous deleterious health outcomes. Despite the high prevalence of sleep disorders, they are widely under-diagnosed at general practice level. AIM: This study aims to investigate the association between demographic and morbidity factors, and self-reported sleep disturbance symptoms. METHODS: A quantitative cross-sectional study design was used. The data collection tool was an anonymous questionnaire consisting of 22 sleep symptoms categorised into four subscales: 1. Insomnia, 2. Daytime Distress, 3. Sleep Disorder, 4. Psychological Distress. Participants were adults ≥18 years of age attending their general practitioner. RESULTS: A total of 281 questionnaires were analysed (70.3% response rate). Participants with a diagnosis of depression and those who experienced low mood 'very frequently' had significantly higher median scores on all four subscales. Those with a body mass index (BMI) >30 kg/m2 had a higher median score on subscale 3, compared to those with lower BMIs. Smokers had higher median scores on subscales 1-3 compared to non-smokers. Participants >65 years of age had lower median scores on all subscales compared to younger participants. Married participants had lower median scores on subscales 1-3 compared to unmarried participants. A total of 37% reported that they would be willing to participate in an overnight sleep study, and 5.3% had been formally diagnosed with a sleep disorder. CONCLUSIONS: A number of factors are significantly associated with sleep disturbance, particularly depression, low mood, elevated BMI and smoking. General practitioners should consider these factors to increase recognition of patients who would benefit from sleep disorder investigation.

Microarray-based comparative genomic indexing of the Cronobacter genus (Enterobacter sakazakii).

Cronobacter (Enterobacter sakazakii) is a recently defined genus consisting of 6 species. To extend our understanding of the genetic relationship between Cronobacter sakazakii BAA-894 and the other species of this genus, microarray-based comparative genomic indexing (CGI) was undertaken to determine the presence/absence of genes identified in the former sequenced genome and to compare 276 selected open reading frames within the different Cronobacter strains. Seventy-eight Cronobacter strains (60 C. sakazakii, 8 C. malonaticus, 5 C. dublinensis, 2 C. muytjensii, 1 C. turicensis, 1 C. genomospecies 1, and 1 Cronobacter sp.) representing clinical and environmental isolates from various geographical locations were investigated. Hierarchical clustering of the CGI data showed that the species grouped as clusters. The 5 C. dublinensis and 2 C. muytjensii strains examined formed distinct species clusters. Moreover, all of the C. sakazakii and 3 of 8 C. malonaticus strains formed a large cluster. The remaining C. malonaticus strains formed a sub-group within a larger cluster that also contained C. turicensis, C. genomospecies 1, and an unknown Cronobacter sp. Cronobacter sakazakii and 3 of 8 C. malonaticus strains could be distinguished from the others within the collection by the presence of 10 fimbrial related genes. Similarly, capsule and/or lipopolysaccharide (LPS) related glycosyltransferases differentiated several of the C. sakazakii strains from each other.

Evaluation of an automated repetitive sequence-based PCR system for subtyping Enterobacter sakazakii.

Enterobacter sakazakii is regarded as a ubiquitous organism that can be isolated from a wide range of foods and environments. Infection in at-risk infants has been epidemiologically linked to the consumption of contaminated powdered infant formula. Preventing the dissemination of this pathogen in a powdered infant formula manufacturing facility is an important step in ensuring consumer confidence in a given brand together with the protection of the health status of a vulnerable population. In this study we report the application of a repetitive sequence-based PCR typing method to subtype a previously well-characterized collection of E. sakazakii isolates of diverse origin. While both methods successfully discriminated between the collection of isolates, repetitive sequence-based PCR identified 65 types, whereas pulsed-field gel electrophoresis identified 110 types showing > or =95% similarity. The method was quick and easy to perform, and our data demonstrated the utility and value of this approach to monitor in-process contamination, which could potentially contribute to a reduction in the transmission of E. sakazakii.

Dissemination of Cronobacter spp. (Enterobacter sakazakii) in a powdered milk protein manufacturing facility.

The microbial contamination of air filters and possible links to contaminated product in a powdered milk protein-processing facility were investigated. Over a 10-month period, seven air filters, the environment, and powdered product were analyzed for the presence of Cronobacter spp. The effects of air filter installation, maintenance, and subsequent dissemination of Cronobacter were investigated. A total of 30 isolates were characterized by pulsed-field gel electrophoresis (PFGE). PFGE revealed the presence of three clonal populations distributed throughout the manufacturing site. This study highlights the need for proper installation of air filters to limit the dissemination of microorganisms into processing sites.

Molecular analysis of the Enterobacter sakazakii O-antigen gene locus.

Nucleotide polymorphism associated with the O-antigen-encoding locus, rfb, in Enterobacter sakazakii was determined by PCR-restriction fragment length polymorphism analysis. Based on the analysis of these DNA profiles, 12 unique banding patterns were detected among a collection of 62 strains from diverse origins. Two common profiles were identified and were designated serotypes O:1 and O:2. DNA sequencing of the 12,500-bp region flanked by galF and gnd identified 11 open reading frames, all with the same transcriptional direction. Analysis of the proximal region of both sequences demonstrated remarkable heterogeneity. A PCR assay targeting genes specific for the two prominent serotypes was developed and applied for the identification of these strains recovered from food, environmental, and clinical samples.

Development of multiple-locus variable-number tandem-repeat analysis for the molecular subtyping of Enterobacter sakazakii.

The genomic content of Enterobacter sakazakii strain ATCC BAA-894 was analyzed for variable-number tandem repeats (VNTRs). In this study we report the development of a multiple-locus VNTR analysis (MLVA) strategy for the subtyping of E. sakazakii. The method is based on a GeneScan analysis of four VNTR loci labeled with multiple fluorescent dyes. This approach was applied to a collection of 112 isolates representing all 16 of the currently defined E. sakazakii biogroups. MLVA successfully discriminated among these isolates and compared favorably with pulsed-field gel electrophoresis. The method was relatively fast and easy to perform. The potential value of MLVA as an epidemiological tool is discussed.

Enterobacter sakazakii an emerging bacterial pathogen with implications for infant health.

Enterobacter sakazakii (E. sakazakii) is an opportunistic pathogen and the aetiological agent in rare but life-threatening cases of meningitis, necrotizing enterocolitis, and sepsis in infants. Among infants, those at greatest risk are neonates (<28 days), particularly those born prematurely or of low birth weight (<2500 g). Consumption of contaminated powdered infant formula (PIF) has been epidemiologically linked with cases of infection. Contamination can occur during the manufacturing process or during postmanufacture reconstitution of formula. Development of rapid, sensitive and specific detection methods will facilitate manufacturers efforts to reduce the occurrence of E. sakazakii in the final powdered product. Furthermore, since PIF is not a sterile product, proper precautions should be taken during handling and reconstitution of formula prior to feeding in order to prevent contamination and proliferation of the bacterium.

Application of pulsed-field gel electrophoresis to characterise and trace the prevalence of Enterobacter sakazakii in an infant formula processing facility.

Enterobacter sakazakii (E. sakazakii) contamination of powdered infant formula (PIF) and its processing environment was monitored between April 2005 and March 2006. The purpose of the monitoring programme was to locate points of contamination, investigate clonal persistence, and identify possible dissemination routes along the processing chain. A total of 80 E. sakazakii isolates were recovered from the manufacturing facility. The overall frequency of isolation of E. sakazakii in intermediate and final product was 2.5%, while specific locations in the processing environment were contaminated at frequencies up to 31%. All E. sakazakii isolates were characterised by pulsed-field gel electrophoresis (PFGE). XbaI macrorestriction digests yielded 19 unique pulse-types that could be grouped into 6 clusters of between 5 and 32 isolates. The formation of large clusters was consistent with the presence of a number of clones in the manufacturing environment. While the majority of isolates were of environmental origin (72.5%), no cluster was confined to one specific location and indistinguishable PFGE profiles were generated from isolates cultured from the manufacturing environment, sampling points along the processing chain and from intermediate and final product. These findings suggest that the manufacturing environment serves as a key route for sporadic contamination of PIF. These data will support the development of efficient intervention measures contributing to the reduction of E. sakazakii in the PIF processing chain.

Detection of Enterobacter sakazakii in dried infant milk formula by cationic-magnetic-bead capture.

Enterobacter sakazakii has been associated with life-threatening infections in premature low-birth-weight infants. Contaminated infant milk formula (IMF) has been implicated in cases of E. sakazakii meningitis. Quick and sensitive methods to detect low-level contamination sporadically present in IMF preparations would positively contribute towards risk reduction across the infant formula food chain. Here we report on the development of a simple method, combining charged separation and growth on selective agar, to detect E. sakazakii in IMF. This protocol can reliably detect 1 to 5 CFU of E. sakazakii in 500 g of IMF in less than 24 h.

Enterobacter sakazakii: an emerging pathogen in powdered infant formula.

Enterobacter sakazakii represents a significant risk to the health of neonates. This bacterium is an emerging opportunistic pathogen that is associated with rare but life-threatening cases of meningitis, necrotizing enterocolitis, and sepsis in premature and full-term infants. Infants aged <28 days are considered to be most at risk. Feeding with powdered infant formula (PIF) has been epidemiologically implicated in several clinical cases. Infants should be exclusively breast-fed for the first 6 months of life, and those who are not should be provided with a suitable breast-milk substitute. PIF is not a sterile product; to reduce the risk of infection, the reconstitution of powdered formula should be undertaken by caregivers using good hygienic measures and in accordance with the product manufacturer's food safety guidelines.

Lack of effect of bromocriptine on ACTH levels in patients with bilateral adrenalectomy for pituitary-dependent Cushing's syndrome.

Bromocriptine lowers plasma ACTH levels in patients with pituitary-dependent Cushing's syndrome. It seemed possible that the drug would also be effective in patients who had been adrenalectomised for the disease. The effect of 5.0 mg of bromocriptine on plasma ACTH and prolactin levels in 13 patients, bilaterally adrenalectomised for pituitary-dependent Cushing's syndrome, was therefore studied. In only one patient was a fall in plasma ACTH observed. This may be of doubtful significance since marked spontaneous fluctuation in plasma ACTH levels were found in the five patients tested. Bromocriptine lowered the plasma prolactin in all of the patients, including two with hyperprolactinaemia.

Acute pancreatitis and vitamin K deficiency in pregnancy.

Two patients with acute pancreatitis in pregnancy are described. In both, bleeding from vitamin K deficiency occurred after the initial attack of pancreatitis and the bleeding tendency was successfully treated with vitamin K.