Severe hypoglycaemia and its association with psychological well-being in Australian adults with type 1 diabetes attending specialist tertiary clinics.

AIM: To investigate severe hypoglycaemia (SH) in adults with type 1 diabetes and its associations with impaired awareness of hypoglycaemia (IAH), clinical, psychological and socio-demographic factors. METHODS: Attendees of three specialist diabetes clinics in Melbourne, Australia completed questions about frequency of SH in the past six months; impaired awareness of hypoglycaemia (Gold score); and measures of general emotional well-being (WHO-5), diabetes-specific positive well-being (subscale of W-BQ28), diabetes-related distress (PAID) and fear of hypoglycaemia (HFS). RESULTS: Of 422 participants (mean ± SD age 37.5 ± 15.0 years; 54% women), 78 (18.5%) reported at least one SH event and 86 (20.5%) had IAH. SH and IAH frequencies were similar at all clinics. In total, 194 SH events were reported, with 10 people experiencing 40% of events. Compared with those without SH, participants with SH had longer diabetes duration, were younger at diabetes onset and more likely to have IAH (p<0.01). Those with SH had greater fear of hypoglycaemia and diabetes-related distress, poorer general emotional well-being, and lower diabetes-specific positive well-being, (p<0.01). There were no associations with age, gender, insulin regimen or HbA1c. CONCLUSIONS: This study has identified that SH and IAH in Australian adults with type 1 diabetes exist at similar levels to those reported in US and European research. SH was significantly associated with IAH and fear of hypoglycaemia. Assessment of hypoglycaemia, IAH and psychological well-being as part of a routine diabetes clinic visit was well accepted by attendees and enabled identification of those who may benefit from medical, educational or therapeutic interventions.

The relationship between family functioning and child and adolescent overweight and obesity: a systematic review.

There is mounting evidence that family functioning is linked to childhood overweight and obesity, and that both of these are associated with health-related behaviours and adverse health outcomes in children and adolescents. This paper systematically examines the peer-reviewed evidence regarding the relationship between child and adolescent overweight and obesity and family functioning. Peer-reviewed literature published between 1990 and 2011 hosted in Scopus, Pub Med or Psyc INFO were searched, in addition to the reference lists of included papers. Twenty-one studies met the selection criteria. Of the 17 identified cross-sectional and longitudinal studies, 12 reported significant associations between family functioning and childhood overweight and obesity. The instruments used to measure family functioning in the identified studies were heterogeneous. Poor family functioning was associated with increased risk of obesity and overweight in children and adolescents, and obese children and adolescents were more likely to come from families with poor family functioning. Aspects of family functioning which were associated with increased risk of child and adolescent obesity included poor communication, poor behaviour control, high levels of family conflict and low family hierarchy values. Half (2/4) of the identified intervention studies showed a significant relationship between family functioning and changes in child weight. The results demonstrate that family functioning is linked to obesity; however, higher level evidence and greater understanding of the mechanisms behind this relationship are required. The results indicate a need for a standardised family functioning measure applicable across populations. The results provide evidence of the value of considering family functioning in childhood obesity research and intervention.

A controlled release pilocarpine buccal insert in the treatment of Sjögren's syndrome.

OBJECTIVES: To assess the efficacy of a novel hydrogel polymer buccal insert containing 5 mg pilocarpine in releasing the pilocarpine in a controlled fashion over a three hour period, and to assess the effects of this on quantitative tear and saliva production and the acceptability of the insert to the patient. DESIGN: This was an open, uncontrolled pilot study for which Ethics Committee approval was obtained prior to starting. Hydrogel buccal inserts containing 5 mg pilocarpine were used three times a day for seven days. SETTING: The Department of Oral Medicine, Glasgow Dental Hospital & School. SUBJECTS: Eight patients with Sjögren's syndrome. MAIN OUTCOME MEASURES: Changes over baseline in (1) Schirmer test, (2) whole saliva flow rate, (3) oral comfort score (VAS), (4) ocular comfort score (VAS), (5) patient acceptability. RESULTS: The buccal inserts successfully released in excess of 85% of their 5 mg pilocarpine load over three hours. There was a general improvement in oral and ocular comfort scores assessed by visual linear analogue scale, and saliva and tear production generally increased. The inserts were well tolerated by all patients except one (who wore dentures). Adverse events were few and none was serious. CONCLUSION: This novel form of buccal pilocarpine delivery demonstrated potential for use in treating patients with Sjögren's syndrome.

Crystallization and preliminary diffraction studies of native and selenomethionine CcmG (CycY, DsbE).

Disulfide-bond (Dsb) proteins are a family of redox proteins containing a Cys-X-X-Cys motif. They are essential for disulfide-bond exchange in the bacterial periplasm and are necessary for the correct folding and function of many secreted proteins. CcmG (DsbE) is a reducing Dsb protein required for cytochrome c maturation. Crystals of Bradyrhizobium japonicum CcmG have been obtained that diffract X-rays to 1.14 A resolution. The crystals are orthorhombic, space group P2(1)2(1)2(1), with unit-cell parameters a = 35.1, b = 48.2, c = 90.2 A. Selenomethionine CcmG was expressed without using a methionine auxotroph or methionine-pathway inhibition and was purified without reducing agents.

Characterization of baboon (Papio hamadryas) milk proteins.

The major proteins of baboon milk were identified as beta-lactoglobulin (beta LG), alpha-lactalbumin (alpha LA), lysozyme, lactoferrin, casein, and albumin by immobiline isoelectric focusing, SDS-PAGE, immunoblotting of gels with rabbit antisera to human alpha LA, lysozyme, and albumin and bovine beta LG and casein, and N-terminal sequencing of proteins blotted from gels. The first 30 N-terminal residues of baboon beta LG are identical to those of macaque (Macaca fasicularis) beta LG except for a (D/N) polymorphism at residue 2. The complete cDNA sequence and derived amino acid composition of beta LG were elucidated using RT-PCR amplification of poly(A)+ mRNA purified from lactating mammary gland. Baboon beta LG consists of 168 amino acid residues (M(r) 20,750) and is the longest beta LG identified to date. beta LG and alpha LA polymorphisms with three (A, B, and C) and two (A and B) variants, respectively, were detected by immobiline IEF, pH 4-6, of individual baboon milk samples at varying stages of lactation.

A rapid, semi-automated method for detection of Galbeta1-4GlcNAc alpha2,6-sialyltransferase (EC 2.4.99.1) activity using the lectin Sambucus nigra agglutinin.

Sialyltransferase activity has traditionally been studied by determining the rate at which the enzyme transfers a labeled donor sugar to an acceptor substrate. These types of assays can be difficult to quantitate, and the separation of untransfered donor sugar from the sialylated acceptor is time-consuming. The biosensor-based method described here is both rapid and semi-automated. The NeuAc-alpha2-6Gal-R-specific lectin Sambucus nigra agglutinin (SNA) immobilized to the carboxymethyl dextran surface of a BIAcore sensor chip was used to detect and measure the formation of the NeuAc-alpha2-6Gal-R moieties. The sialyltransferase assays were carried out using modified protocols based on the method described in Rearick, J.I., Sadler, J.E., Paulson, J.C., and Hill, R.L. (1979) Enzymatic characterization of betaD-galactoside alpha2-3 sialyltransferase from porcine submaxillary gland. J. Biol. Chem., 254, 4444-4451. The complete assay mixture was simply diluted before injection into the instrument. All injections were performed automatically using the robotics of the BIAcore instrument. Using this technique it is possible to detect product from 0.4 microU of commercial Galbeta1-4GlcNAc alpha2,6-sialyltransferase (EC 2.4.99.1) (ST6Gal I). One unit of sialyltransferase is defined as the quantity that will transfer 1 micromol of N-acetylneuraminic acid from cytidine monophosphate (CMP)-N-acetylneuraminic acid to asialofetuin per min at pH 6.5 and 37 degrees C. The method described here requires as little as 10 microl total assay volume, thus reducing the consumption of reagents. In addition, the sample is completely recoverable from the sensor chip surface, which allows for downstream analysis of the reaction product if desired. This method eliminates the need for labeled donor and acceptor molecules and does not require the separation of the substrates from the product before analysis. Although some kinetic properties of the enzyme can be estimated using this method, further development and validation is required. The method is most useful in determining qualitative estimates of ST6Gal I activity in tissue extracts and in characterizing the production of enzymes in cultured cell systems. The use of a microtiter plate assay format enables the rapid screening of multiple fractions for sialyltransferase activity.

Host-specificity of monogenean (platyhelminth) parasites: a role for anterior adhesive areas?

Monogeneans (flatworms) are among the most host-specific of parasites in general and may be the most host-specific of all fish parasites. Specificity, in terms of a restricted spatial distribution within an environment, is not unique to parasites and is displayed by some fungi, insects, birds, symbionts and pelagic larvae of free-living marine invertebrates. The nature of cues, how "habitats" are recognised and how interactions between partners are mediated and maintained is of interest across these diverse "associations". We review some experiments that demonstrate important factors that contribute to host-specificity at the level of infective stages (larvae of oviparous monogeneans; juveniles of viviparous gyrodactylids) and adult parasites. Recent research on immune responses by fish to monogenean infections is considered. We emphasise the critical importance of host epidermis to the Monogenea. Monogeneans live on host epidermis, they live in its products (e.g. mucus), monopisthocotyleans feed on it, some of its products are "attractants" and it may be an inhospitable surface because of its immunological activity. We focus attention on fish but reference is made to amphibian hosts. We develop the concept for a potential role in host-specificity by the anterior adhesive areas, either the specialised tegument and/or anterior secretions produced by monogeneans for temporary but firm attachment during locomotion on host epithelial surfaces. Initial contact between the anterior adhesive areas of infective stages and host epidermis may serve two important purposes. (1) Appropriate sense organs or receptors on the parasite interact with a specific chemical or chemicals or with surface structures on host epidermis. (2) A specific but instant recognition or reaction occurs between component(s) of host mucus and the adhesive(s) secreted by monogeneans. The chemical composition of fish skin is known to be species-specific and our preliminary analysis of the chemistry of some monogenean adhesives indicates they are novel proteins that display some differences between parasite families and species.

Preparation and characterization of polyclonal antibodies against human chaperonin 10.

Early pregnancy factor (EPF) has been identified as an extracellular homologue of chaperonin 10 (Cpn10), a heat shock protein that functions within the cell as a molecular chaperone. Here, we report the production of polyclonal antibodies directed against several different regions of the human Cpn10 molecule and their application to specific protein quantitation and localization techniques. These antibodies will be valuable tools in further studies to elucidate the mechanisms underlying the differential spatial and temporal localization of EPF and Cpn10 and in studies to elucidate structure and function.

Genetic analysis of mutagenesis in aging Escherichia coli colonies.

Bacteria live in unstructured and structured environments, experiencing feast and famine lifestyles. Bacterial colonies can be viewed as model structured environments. SOS induction and mutagenesis have been observed in aging Escherichia coli colonies, in the absence of exogenous sources of DNA damage. This cAMP-dependent mutagenesis occurring in Resting Organisms in a Structured Environment (ROSE) is unaffected by a umuC mutation and therefore differs from both targeted UV mutagenesis and recA730 (SOS constitutive) untargeted mutagenesis. As a recB mutation has only a minor effect on ROSE mutagenesis it also differs from both adaptive reversion of the lacI33 allele and from iSDR (inducible Stable DNA Replication) mutagenesis. Besides its recA and lexA dependence, ROSE mutagenesis is also uvrB and polA dependent. These genetic requirements are reminiscent of the untargeted mutagenesis in lambda phage observed when unirradiated lambda infects UV-irradiated E. coli. These mutations, which are not observed in aging liquid cultures, accumulate linearly with the age of the colonies. ROSE mutagenesis might offer a good model for bacterial mutagenesis in structured environments such as biofilms and for mutagenesis of quiescent eukaryotic cells.

Teacher initiated improvement of asthma policy in schools.

OBJECTIVE: To assess whether information about asthma acquired by individual teachers, following attendance at an inservice seminar, would be communicated to other school staff, and whether policies about asthma management would be subsequently developed in schools. METHODOLOGY: A telephone interview was conducted of 50 primary school teachers in the Hunter Region who were selected randomly from those who had represented their school at an inservice seminar about asthma. RESULTS: All 48 respondents had provided feedback to their colleagues, mostly at a staff meeting. After the seminar the number of schools with a written policy about the management of asthma had increased from 1 to 20. Initially the school's first aid kits contained a bronchodilator inhaler in 25 schools (increasing to 43 after the seminar), a spacer device in six schools (increasing to 47) and written guidelines for management of an asthma attack in seven schools (increasing to 32). Teachers from four schools indicated that all school staff knew how to recognize an acute attack prior to the seminar compared with 22 schools afterwards. Before the seminar 18 schools asked parents to supply written instructions for the management of their children at school but this increased to 44 after the seminar. CONCLUSIONS: A programme of one-off seminars attended by one teacher from each primary school resulted in large increases in the proportion of schools with appropriate policies for the management of asthma in the school environment.

Inversions with deletions and duplications.

Complex mutational events, including de novo inversion with deletion and duplication of sequence, have been observed but are difficult to model. We propose that nascent leading-strand misalignment upon the lagging-strand template during DNA replication can result in the inversion of sequence. The positioning of this misalignment and of the realignment of the leading strand back into the leading-strand template will determine if the inversion is accompanied by deletion and duplication of sequence. We suggest that such strand misalignment-realignment events may occur at the replication fork during concurrent DNA replication.

Parental asthma knowledge: its association with readmission of children to hospital.

OBJECTIVE: To assess whether poor parental knowledge about asthma was a risk factor for readmission of their children to hospital. METHODOLOGY: Parents of children who were admitted to the paediatric facility at Newcastle Mater and John Hunter Hospitals were administered a previously validated asthma knowledge questionnaire with a maximum possible score of 31. RESULTS: The mean parental knowledge scores for three groups of children were determined: those with a previous diagnosis of asthma being readmitted to hospital (170 children, mean score 18.3); those with a previous diagnosis of asthma being admitted for the first time (114 children, mean score 17.2); and children being admitted with their first attack of asthma (65 children, mean score 15.6). There was a significant difference in parental asthma knowledge between the first and third group (P < 0.001) but not between the first and second group. A linear regression model with knowledge score as an outcome found that increased knowledge was associated with previously diagnosed asthma, higher parental occupation and no smoker in the home. CONCLUSIONS: Knowledge was poor in all groups. Readmission rate was related to the severity of the child's asthma.

Editing DNA replication and recombination by mismatch repair: from bacterial genetics to mechanisms of predisposition to cancer in humans.

A hereditary form of colon cancer, hereditary non-polyposis colon cancer (HNPCC), is characterized by high instability of short repeated sequences known as microsatellites. Because the genes controlling microsatellite stability were known in bacteria and yeast, as was their evolutionary conservation, the search for human genes responsible for HNPCC became a 'targeted' search for known sequences. Mismatch-repair deficiency in bacteria and yeast produces multiple phenotypes as a result of its dual involvement in the editing of both replication errors and recombination intermediates. In addition, mismatch-repair functions are specialized in eukaryotes, characterized by specific mitotic (versus meiotic) functions, and nuclear (versus mitochondrial) localization. Given the number of phenotypes observed so far, we predict other links between mismatch-repair deficiency and human genetic disorders. For example, a similar type of sequence instability has been found in HNPCC tumours and in a number of neuro-muscular genetic disorders. Several human mitochondrial disorders display genomic instabilities reminiscent of yeast mitochondrial mismatch-repair mutants. In general, the process of mismatch repair is responsible for the constant maintenance of genome stability and its faithful transmission from one generation to the next. However, without genetic alteration, species would not be able to adapt to changing environments. It appears that nature has developed both negative and positive controls for genetic diversity. In bacteria, for example, an inducible system (sos) exists which generates genetic alterations in response to environmental stress (e.g. radiation, chemicals, starvation). Hence, the cost of generating diversity to adapt to changing conditions might be paid as sporadic gene alterations associated with disease.

Two hour seminar improves knowledge about childhood asthma in school staff.

Teachers and support staff are often called upon to manage asthma at school but may have little knowledge and understanding of the condition. The aim of this study was to determine whether a short seminar would improve knowledge in non-health professionals. Seventy school staff attended one of five 2 h seminars presented by an asthma educator. Participants completed the Newcastle Asthma Knowledge Questionnaire (AKQ) before, and 2 months after, the seminar. The mean initial score (maximum possible 31) was 16.4 (range 4-26), increasing to 23 (range 13-29, P = 0.0001). There was a large increase in knowledge about symptoms, pathophysiology, preventive medications and side effects of medications. Initially almost all of the participants had deficiencies in knowledge about reliever medications and the management of exercise-induced asthma. After the seminar, knowledge in these areas was still poor, with only one-third of the participants answering them correctly. The results have been used to modify the content of the 2 h seminar with increased emphasis on those areas in which deficiencies in knowledge were demonstrated. No data are available yet as to whether this improved knowledge has improved management.

Headache and asthma.

The aim of this study was to investigate the association between headache and asthma, bronchodilators and atopy in school children. A cross-sectional survey of all primary school children was conducted in two towns near Newcastle, New South Wales, Australia; one in the vicinity of two coal-fired power stations, the other free of outdoor industrial air pollution. The main outcome measures were frequent headache, wheezing, bronchial reactivity, use of bronchodilators and atopy. Eight hundred and fifty-one primary school children aged 5-12 years participated (92% response rate). Twenty-three per cent of the children were reported to have had a history of frequent headache. Crude odds ratios indicated that the odds of frequent headache was significantly higher in children with asthma and atopy and where there was a smoker in the home, but that there was no association between frequent headache and use of bronchodilators or the sex of the child or socio-economic status measured as father's occupation. Stepwise logistic regression with frequent headache as the outcome of interest showed that, after adjusting for age and smoking in the home, the odds ratio for asthma (defined as current wheeze) was 3.24 (95% confidence interval [CI] 2.19-4.77). The similarly adjusted odds ratio for asthma defined as bronchial hyperreactivity (BHR) was 1.60 (95% CI 1.09-2.37). Atopy was not statistically significantly associated with headache for either model. Asthma (defined as wheeze or BHR) is an independent risk factor for frequent headache. The relationship between headache and asthma is an association with bronchial hyperresponsiveness rather than atopy.