Predictors of type 2 diabetes remission in the Diabetes Remission Clinical Trial (DiRECT).

AIM: To identify predictors of type 2 diabetes remission in the intervention arm of DiRECT (Diabetes Remission Clinical Trial). METHODS: Participants were aged 20-65 years, with type 2 diabetes duration of <6 years and BMI 27-45 kg/m2 , and were not receiving insulin. Weight loss was initiated by total diet replacement (825-853 kcal/day, 3-5 months, shakes/soups), and weight loss maintenance support was provided for 2 years. Remissions (HbA1c <48 mmol/mol [<6.5%], without antidiabetes medications) in the intervention group (n = 149, mean age 53 years, BMI 35 kg/m2 ) were achieved by 68/149 participants (46%) at 12 months and by 53/149 participants (36%) at 24 months. Potential predictors were examined by logistic regression analyses, with adjustments for weight loss and effects independent of weight loss. RESULTS: Baseline predictors of remission at 12 and 24 months included being prescribed fewer antidiabetes medications, having lower triglyceride and gamma-glutamyl transferase levels, and reporting better quality of life with less anxiety/depression. Lower baseline HbA1c was a predictor at 12 months, and older age and male sex were predictors at 24 months. Being prescribed antidepressants predicted non-remission. Some, but not all effects were explained by weight loss. Weight loss was the strongest predictor of remission at 12 months (adjusted odds ratio per kg weight loss 1.24, 95% CI 1.14, 1.34; P < 0.0001) and 24 months (adjusted odds ratio 1.23, 95% CI 1.13, 1.35; P <0.0001). Weight loss in kilograms and percentage weight loss were equally good predictors. Early weight loss and higher programme attendance predicted more remissions. Baseline BMI, fasting insulin, fasting C-peptide and diabetes duration did not predict remission. CONCLUSIONS: Other than weight loss, most predictors were modest, and not sufficient to identify subgroups for which remission was not a worthwhile target.

Views, experience and adherence among pregnant women with gestational diabetes participating in a weight loss study (WELLBABE).

AIMS: To investigate the views and experience of pregnant women newly diagnosed with gestational diabetes mellitus participating in a 1200 kcal/day diet to achieve moderate weight loss (the WELLBABE study), and to explore barriers to and facilitators of adherence. METHODS: Twelve participants engaged in semi-structured interviews after completion of the 4-week diet. An interview schedule was devised using open-ended questions guided by the Theoretical Domains Framework. Transcript responses were analysed thematically. RESULTS: Participants were anxious about their diagnosis of gestational diabetes, but concerns related to dieting in pregnancy were allayed by reassurance from the research team. Participants expected health benefits, improved knowledge and support from enrolling on the study. The participants' primary motivator to diet adherence was their baby's wellbeing. Other facilitatory factors included improving their own health and reducing any future risk of diabetes. Trying to provide reliable results and receiving extra care also facilitated adherence. Partners, friends and family were an important source of social support and no barrier caused by concern about weight loss in pregnancy was encountered. Observed and experienced physical changes and feedback from the research team positively reinforced adherence. The main barrier was that learning new skills was initially time-consuming. CONCLUSIONS: Weight loss was acceptable to women with gestational diabetes provided with clear information about likely benefit. A randomized controlled trial of this intervention is now required, employing clear information and feedback of glycaemic benefit to facilitate efficacy.

Can type 2 diabetes be reversed and how can this best be achieved? James Lind Alliance research priority number one.

The James Lind Alliance, in association with Diabetes UK, conducted a survey of people with Type 2 diabetes to establish their priorities for research. The number one research priority was found to be 'Can Type 2 diabetes be cured or reversed, what is the best way to achieve this, and is there a point beyond which the condition cannot be reversed?'. The present review summarizes the current understanding of weight loss-induced reversal of Type 2 diabetes. It considers the diagnostic criteria for remission and describes the clinical features of post-diabetes. It is of great importance to recognize these, as post-diabetes differs considerably from the high cardiovascular risk state of prediabetes. Current data demonstrate long-term stable ß-cell function, providing weight regain is prevented. If an individual, having previously demonstrated susceptibility to Type 2 diabetes, returns to their previous weight then recurrence of the condition is certain. Appropriate use of the terms 'reversal' and 'remission' is discussed, with emphasis that the word 'cure' is inappropriate. Evidence-based means of achieving and maintaining remission of Type 2 diabetes are described, together with a summary of the information on the steadily diminishing chance of achieving reversal with increasing duration of Type 2 diabetes.

Epizootics of Streptococcus agalactiae infection in captive rays from Queensland, Australia.

The aim of this study was to describe two epizootics of high mortalities from infection with Streptococcus agalactiae, occurring in captive rays held in a marine display aquarium in south-east Queensland, Australia, in 2009 and 2010. Five different species of rays were affected, including mangrove whiprays (Himantura granulata), estuary rays (Dasyatis fluviorum), eastern shovelnose rays (Aptychotrema rostrata), white-spotted eagle rays (Aetobatus narinari) and blue-spotted mask rays (Neotrygon kuhlii). This report describes the history of both epizootics including collection, quarantine and husbandry of rays, the disease epizootics, clinico-pathological features of the disease, antimicrobial therapy, autogenous vaccine production, and laboratory studies including clinical and histopathology, bacteriology, PCR, molecular serotyping and sequencing of the bacterium S. agalactiae.

Infection and pathology in Queensland grouper, Epinephelus lanceolatus, (Bloch), caused by exposure to Streptococcus agalactiae via different routes.

Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, (Bloch), have been found dead in NE Australia. In some cases, Streptococcus agalactiae (Group B Streptococcus, GBS) was isolated. At present, a GBS isolate from a wild grouper case was employed in experimental challenge trials in hatchery-reared Queensland grouper by different routes of exposure. Injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimming. Death occurred in, and GBS was re-isolated from, 98% fish injected and was detected by PCR in brain, head kidney and spleen from all fish, regardless of challenge dose. Challenge by immersion resulted in lower morbidity with a clear dose response. Whilst infection was established via oral challenge by admixture with feed, no mortality occurred. Histology showed pathology consistent with GBS infection in organs examined from all injected fish, from fish challenged with medium and high doses by immersion, and from high-dose oral challenge. These experimental challenges demonstrated that GBS isolated from wild Queensland grouper reproduced disease in experimentally challenged fish and resulted in pathology that was consistent with that seen in wild Queensland grouper infected with S. agalactiae.

Isolation and characterization of koi herpesvirus (KHV) from Indonesia: identification of a new genetic lineage.

Koi herpesvirus (KHV) is the aetiological agent of an emerging disease (KHVD) associated with mass mortalities in koi and common carp and reported from at least 30 countries. We report the first isolation of KHV from koi and common carp in Indonesia and initial characterization of the isolates. Clinical signs, histopathology and virion morphology are similar to those of isolates from other countries. Phylogenetic analyses using the thymidine kinase gene amplified from each isolate and from carp tissue samples collected from KHVD outbreaks throughout Indonesia indicated that the Indonesian isolates are more closely related to the Asian than the European KHV lineage. Sequence analysis of two other variable regions between ORF29 and ORF31 (marker I) and near the start of ORF 133 (marker II) indicated that all Indonesian isolates displayed a marker I allele (I(++)) previously identified only in isolates of the Asian lineage. However, in the marker II region, all Indonesian isolates displayed the II(-) allele, which has been reported previously only amongst isolates of the European lineage, and nine of these displayed a mixed genotype (II(+)II(-)). The I(++)II(-) genotype has not been reported previously and appears to represent a new intermediate lineage that may have emerged in Indonesia.

Clinical validation of an autoantibody test for lung cancer.

BACKGROUND: Autoantibodies may be present in a variety of underlying cancers several years before tumours can be detected and testing for their presence may allow earlier diagnosis. We report the clinical validation of an autoantibody panel in newly diagnosed patients with lung cancer (LC). PATIENTS AND METHODS: Three cohorts of patients with newly diagnosed LC were identified: group 1 (n = 145), group 2 (n = 241) and group 3 (n = 269). Patients were individually matched by gender, age and smoking history to a control individual with no history of malignant disease. Serum samples were obtained after diagnosis but before any anticancer treatment. Autoantibody levels were measured against a panel of six tumour-related antigens (p53, NY-ESO-1, CAGE, GBU4-5, Annexin 1 and SOX2). Assay sensitivity was tested in relation to demographic variables and cancer type/stage. RESULTS: The autoantibody panel demonstrated a sensitivity/specificity of 36%/91%, 39%/89% and 37%/90% in groups 1, 2 and 3, respectively, with good reproducibility. There was no significant difference between different LC stages, indicating that the antigens included covered the different types of LC well. CONCLUSION: This assay confirms the value of an autoantibody panel as a diagnostic tool and offers a potential system for monitoring patients at high risk of LC.

Bacterial diversity of the digestive gland of Sydney rock oysters, Saccostrea glomerata infected with the paramyxean parasite, Marteilia sydneyi.

AIMS: To determine whether the infestation by the protozoan paramyxean parasite, Marteilia sydneyi, changes the bacterial community of the digestive gland of Sydney rock oysters, Saccostrea glomerata. METHODS AND RESULTS: Six 16S rDNA clone libraries were established from three M. sydneyi-infected and three un-infected oysters. Restriction enzyme analysis followed by sequencing representative clones revealed a total of 23 different operational taxonomic units (OTUs) in un-infected oysters, comprising the major phyla: Firmicutes, Proteobacteria, Cyanobacteria and Spirocheates, where the clone distribution was 44, 36, 7 and 5%, respectively. Close to half of the OTUs are not closely related to any other hitherto determined sequence. In contrast, S. glomerata infected by M. sydneyi had only one OTU present in the digestive gland. Phylogenetic analysis of the 16S rDNA sequence reveals that this dominant OTU, belonging to the alpha-Proteobacteria, is closely related to a Rickettsiales-like prokaryote (RLP). CONCLUSIONS: The microbiota of the digestive gland of Sydney rock oysters is changed by infection by M. sydneyi, becoming dominated by a RLP, and generally less diverse. The bacterial community of un-infected S. glomerata differs from previous studies in that we identified the dominant taxa as Firmicutes and alpha-Proteobacteria, rather than heterotrophic gamma-Proteobacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first culture-independent study of the microbiota of the digestive glands of edible oysters to the species level. The commercial viability of the Sydney rock oyster industry in Australia is currently threatened by Queensland Unknown disease and the changes in the bacterial community of S. glomerata corresponding with infection by M. sydneyi sheds further light on the link between parasite infection and mortality in this economically damaging disease.

Isolation of a novel strain of Bacillus pumilus from penaeid shrimp that is inhibitory against marine pathogens.

A bacterium was isolated from the mid-gut of healthy black tiger shrimp, Penaeus monodon, based on a large zone of inhibition in mixed culture on solid medium. The isolate was a Gram-positive, motile spore former, with an optimum pH range for growth in tryptone soya broth containing 2% NaCl of between pH 6 and 9. The bacterium was highly salt tolerant with concentrations between 0% and 8% having no detrimental effect on growth. The isolate was identified as Bacillus pumilus based on physiological capabilities using the API50CHB and Biolog systems. Amplification and sequencing of the 16S rRNA gene followed by phylogenetic analysis confirmed its identity. The Bacillus pumilus isolate was strongly inhibitory against the marine bacterial pathogens Vibrio alginolyticus, V. mimicus and V. harveyi, and weakly inhibitory against V. parahaemolyticus in cross-streaking assays on solid medium. The organism was marginally self-inhibitory, and inhibited B. licheniformis and B. subtilis. The suitability of the B. pumilus isolate for use as a probiotic in farmed shrimp was further supported by the absence of any of the known B. cereus enterotoxin genes. Based on these in vitro results, in vivo safety and efficacy trials are underway to determine suitability of the novel strain as a commercial probiotic.

Autoantibodies in lung cancer: possibilities for early detection and subsequent cure.

BACKGROUND: People with lung cancer usually present at a late stage in the course of their disease when their chances of long-term survival are low. At present there is little to offer for early diagnosis, even in those at high risk of developing the disease. Autoantibodies have been shown to be present in the circulation of people with various forms of solid tumour before cancer-associated antigens can be detected, and these molecules can be measured up to 5 years before symptomatic disease. OBJECTIVE: To assess the potential of a panel of tumour-associated autoantibody profiles as an aid to other lung cancer screening modalities. METHODS: Plasma from normal controls (n = 50), patients with non-small cell lung cancer (n = 82) and patients with small cell lung cancer (n = 22) were investigated for the presence of autoantibodies to p53, c-myc, HER2, NY-ESO-1, CAGE, MUC1 and GBU4-5 by enzyme-linked immunosorbent assay. RESULTS: Raised levels of autoantibodies were seen to at least 1/7 antigens in 76% of all the patients with lung cancer plasma tested, and 89% of node-negative patients, with a specificity of 92%. There was no significant difference between the detection rates in the lung cancer subgroups, although more patients with squamous cell carcinomas (92%) could be identified. CONCLUSION: Measurement of an autoantibody response to one or more tumour-associated antigens in an optimised panel assay may provide a sensitive and specific blood test to aid the early detection of lung cancer.

Wildlife as reservoirs for parasites infecting commercial species: host specificity and a redescription of Kudoa amamiensis from teleost fish in Australia.

Parasites of the genus Kudoa (Phylum Myxozoa) have long been known to cause considerable losses to finfish aquaculture. One such parasite species, Kudoa amamiensis, causes unsightly white cysts in the skeletal muscle of yellowtail kingfish, Seriola quinqueradiata, in Japan rendering the fillets unmarketable. The authors who characterized K. amamiensis, Egusa & Nakajima, 1980, hypothesized that yellowtail kingfish, as non-natives to the area, were accidental hosts of the parasite and that it normally infects native reef fish (damselfish, Family Pomacentridae). Since then, we have found parasites that are consistent with the description of K. amamiensis in two species of damselfish and one species of carangid fish in Australia, and it has been recorded previously in another species of reef-associated fish. Our morphometric, histological and DNA results suggest that these specimens are K. amamiensis, and are new host records for that species. Furthermore, our observations show that reef fish may act as a reservoir of myxozoan infection for commercial species, and as such should be considered an infection pathway for species in aquaculture.

X-ray and neutron scattering studies of the hydration structure of alkali ions in concentrated aqueous solutions.

The presence of ions in water provides a rich and varied environment in which many natural processes occur with important consequences in biology, geology and chemistry. This article will focus on the structural properties of ions in water and it will be shown how the 'difference' methods of neutron diffraction with isotopic substitution (NDIS) and anomalous X-ray diffraction (AXD) can be used to obtain direct information regarding the radial pair distribution functions of many cations and anions in solution. This information can subsequently be used to calculate coordination numbers and to determine ion-water conformation in great detail. As well as enabling comparisons to be made amongst ions in particular groups in the periodic table, such information can also be contrasted with results provided by molecular dynamics (MD) simulation techniques. To illustrate the power of these 'difference' methods, reference will be made to the alkali group of ions, all of which have been successfully investigated by the above methods, with the exception of the radioactive element francium. Additional comments will be made on how NDIS measurements are currently being combined with MD simulations to determine the structure around complex ions and molecules, many of which are common in biological systems.

Novel behaviour and structure of new glasses of the type Ba-Al-O and Ba-Al-Ti-O produced by aerodynamic levitation and laser heating.

Novel barium aluminate (BaAl(2)O(4)) and barium alumino-titanate (BaAl(2)TiO(6)) glasses have been produced by aerodynamic levitation and laser heating. BaAl(2)O(4) forms a clear and colourless glass under containerless and rapid quenching conditions. Under similar rapid quenching conditions BaAl(2)TiO(6) forms an opaque and black glass, while under slower and controlled quenching conditions it is possible to form a clear and colourless glass. The formation of the opaque or clear glass is reversible and purely dependent on the quench rate used. By slowing the quench rate further, it is possible to produce a milky glass suggestive of liquid-liquid phase separation in the liquid before glassification. High-energy x-ray diffraction experiments confirm the glassy state of these materials and show coordination structures and bond distances similar to their crystalline analogues.

An anomalous x-ray diffraction study of the hydration structures of Cs+ and I- in concentrated solutions.

Anomalous x-ray diffraction experiments were carried out on concentrated aqueous solutions of sodium iodide (6 molal) and cesium iodide (3 molal). Data were gathered at two energies below the absorption edges of the Cs+ and I- ions in order to avoid contributions from fluorescence. The statistics and quality of the raw data were improved by the use of a focusing analyzer crystal. Differences were taken between the data sets and used to calculate the hydration structures of Cs+ and I-. The structures found are more complex than anticipated for such large ions with relatively low charge densities and show evidence of ion-pair formation in both solutions. A two-Gaussian fit to the Cs+ data gives information about the Cs+-O and Cs+-I- correlations. The central position of the Gaussian representing the Cs+-O was fixed at 3.00 A, that is, the maximum of this contribution. The other parameters were allowed to vary freely, giving a Cs+-I- distance of 3.84+/-0.05 A and coordination numbers of 7.9 and 2.7, respectively, for the Cs+-O and Cs+-I- correlations. The results on the structure of I- in the 6 molal NaI aqueous solution were also fitted to a model based on Gaussians; this gives correlations for I- -O and I- -Na+ at 3.17+/-0.06 and 3.76+/-0.06 A with respective coordination numbers of 8.8 and 1.6. The structure of I- in the 3 molal CsI solution shows overlapping contributions due to I- -H, I- -O, and I- -Cs+. The best Gaussian fit gives two peaks centered at 3.00+/-0.08 and 3.82+/-0.04 A and shows that the latter two correlations are unresolved. The hydration structures are compared with those of other alkali and halide ions. The results are also found to be in good agreement with those obtained from standard x-ray diffraction and computer simulation.

Iron acquisition mechanisms of Flavobacterium psychrophilum.

Forty strains of Flavobacterium psychrophilum were tested for the production of siderophores using the universal Chrome Azurol S (CAS) assay. The majority of the strains (85%) were CAS positive (CAS+) and some (15%) were CAS negative (CAS-). The cryptic plasmid pCP1 was carried by all positive strains and was lacking from negative strains. While a weak catechol reaction was detectable in CAS+ culture supernatants, the CAS reaction was, to some extent, heat sensitive, questioning whether the positive reaction was caused only by siderophores. The ability to grow in vitro under iron-restricted conditions did not correlate with the CAS reactivity, as growth of both CAS+ and CAS- strains was similarly impaired under iron restriction induced by 2,2 dipyridyl. Suppressed growth under these conditions was restored by addition of FeCl3, haemoglobin and transferrin for both CAS+ and CAS- strains.

Update on bacterial vaccines: Photobacterium damselae subsp. piscicida.

Photobacterium damselae subsp. piscicida is the causative agent of pasteurellosis in wild and farmed marine fish worldwide. Although serologically homogeneous, recent molecular advances have led to the discovery of distinct genetic clades, depending on geographical origin. Further details of the strategies for host colonisation have arisen including information on the role of capsule, susceptibility to oxidative stress, confirmation of intracellular survival in host epithelial cells, and induced apoptosis of host macrophages. This improved understanding has given rise to new ideas and advances in vaccine technologies, which are reviewed in this paper.

The hydration structure of guanidinium and thiocyanate ions: implications for protein stability in aqueous solution.

Neutron diffraction experiments were carried out on aqueous solutions containing either guanidinium or thiocyanate ions. The first-order difference method of neutron diffraction and isotopic substitution was applied, and the hydration structures of two of nature's strongest denaturant ions were determined. Each ion is shown to interact weakly with water: Guanidinium has no recognizable hydration shell and is one of the most weakly hydrated cations yet characterized. Hydration of thiocyanate is characterized by a low coordination number involving around one hydrogen-bonded water molecule and approximately two water molecules weakly interacting through "hydration bonds." The weak hydration of these denaturant ions strongly supports suggestions that a major contribution to the denaturant effect is the preferential interaction of the denaturant with the protein surface. By contrast, solute species such as many sugars and related polyols that stabilize proteins are strongly hydrated and are thus preferentially retained in the bulk solvent and excluded from the protein surface.