"I have got diabetes!" - interviews of patients newly diagnosed with type 2 diabetes.

BACKGROUND: To be diagnosed with type 2 diabetes is a challenge for every patient. There are previous studies on patients' experience in general but not addressing the increased cardiovascular risk and multifactorial treatment. The aim of this study was to explore the thoughts, experiences and reactions of newly diagnosed patients with diabetes to this diagnosis and to the risk of developing complications. METHODS: Ten adults (7 men/3 women, aged 50-79) diagnosed with type 2 diabetes within the last 12 months were interviewed at a primary health care center in Sweden. An interview guide was used in the semi-structured interviews that were transcribed verbatim. The analysis was qualitative and inspired by systematic text condensation (Malterud). The text was read several times and meaning units were identified. Related meaning units were sorted into codes and related codes into categories during several meetings between the authors. Finally, the categories were merged and formed themes. RESULTS: We defined three main themes: Reaction to diagnosis, Life changes and Concerns about the future. Most patients reacted to the diagnosis without intensive feelings. Lifestyle changes were mainly accepted but hard to achieve. The patients' major concerns for the future were the consequences for daily life (being able to drive and read) and concerns for relatives rather than anxieties regarding medical issues such as laboratory tests. There were considerable differences in how much patients wanted to know about their future risks. CONCLUSIONS: The results of this study might help to focus doctor-patient communication on issues highlighted by the patients and on the importance of individualizing information and recommendations for each patient.

C-peptide concentration, mortality and vascular complications in people with Type 2 diabetes. The Skaraborg Diabetes Register.

AIM: To study the prognosis of patients with newly diagnosed Type 2 diabetes in primary care in relation to their baseline C-peptide concentration. METHODS: C-peptide concentrations were determined in 399 patients aged < 65 years with newly diagnosed Type 2 diabetes using the Skaraborg Diabetes Register, Sweden. Data on cardiovascular complications and death were extracted from national registers and a local study of retinopathy. Statistical analyses were performed using Cox regression. RESULTS: An analysis of C-peptide concentrations in quartiles, after adjusting for confounders, showed that patients in the highest quartile had a 2.75-fold higher risk of death from all causes compared with those in the lowest quartile (CI 1.17-6.47). By contrast, C-peptide concentration was not associated with the incidence of cardiovascular events or the development of retinopathy. CONCLUSIONS: Measurement of C-peptide concentration at diagnosis could help identify patients who are at high risk and who presumably would benefit from more intensive treatment.

Association between copeptin and declining glomerular filtration rate in people with newly diagnosed diabetes. The Skaraborg Diabetes Register.

AIMS: Copeptin has shown association with development of chronic kidney disease (CKD) in people with diabetes. Early detection of individuals having the highest risk could help avoid this complication. Therefore we decided to study copeptin concentrations and estimated glomerular filtration rate (eGFR) retrospectively in people with newly diagnosed diabetes. METHODS: People with newly diagnosed type 2 diabetes in 1996-1998 from Skaraborg Diabetes Register (SDR) were reinvestigated in 2008-2010. Copeptin concentration at the time of diagnosis was determined. Creatinine and cystatin C were used for determination of eGFR at baseline and at reinvestigation (n=161). Data on cardiovascular complications were extracted from national registers. Analyzes were done with logistic regression. RESULTS: From baseline to follow up eGFR decreased with 33ml. Twenty-nine individuals (18.1%) developed CKD stage 3. There was a significant association between elevated copeptin concentrations and development of CKD stage 3 (OR=1.78, 95% CI=1.01-3.16). When adjusting for GFR at baseline the association between copeptin and GFR decline was borderline significant (OR=1.79, 95% CI=0.99-3.25, p=0.055). CONCLUSIONS: Determination of copeptin may early identify people with diabetes and high risk for CKD. To prevent complications for these individuals aggressive treatment should be discussed.

Screening methods for the detection of antibiotic residues in slaughter animals: comparison of the European Union Four-Plate Test, the Nouws Antibiotic Test and the Premi®Test (applied to muscle and kidney).

Microbial growth inhibition tests are widely used as the primary screening approach for the detection of antibiotic residues in slaughter animals. In this study we evaluated and compared the performance of the European Union Four-Plate Test (EU4pt), the Nouws Antibiotic Test (NAT), and a commercial ampoule test, the PremiTest (applied to both muscle and kidney), by parallel analysis of 735 slaughter animals. The EU4pt only showed significant inhibition with two muscle samples containing 305 µg kg(-1) doxycycline and 648 µg kg(-1) tulathromycin, while an maximum residue limit (MRL) violation of 1100 µg kg(-1) sulfamethazine remained unnoticed. PremiTest-muscle only detected the sulfamethazine containing sample, all other (1.1%) suspect samples appeared false-positive results. The same test applied to kidney yielded 4.1% suspect samples, while the NAT screening (based on analysis of renal pelvis fluid) showed 4.9% suspect results. The vast majority of these samples contained tetracycline and/or aminoglycoside residues. PremiTest-kidney appeared to be more sensitive to aminoglycosides than the NAT screening, which failed to detect an MRL violation of 870 µg kg(-1) gentamicin in kidney. Detection of less than MRL levels of tetracycline residues by the NAT proved its suitability for this residue group. Whether PremiTest is sufficiently sensitive for accurate tetracycline detection in kidney remains doubtful, although changing over to kidney definitely improved the suitability of PremiTest for the detection of residues in slaughter animals.

Application of a luminescent bacterial biosensor for the detection of tetracyclines in routine analysis of poultry muscle samples.

Tetracyclines are extensively used in veterinary medicine. For the detection of tetracycline residues in animal products, a broad array of methods is available. Luminescent bacterial biosensors represent an attractive inexpensive, simple and fast method for screening large numbers of samples. A previously developed cell-biosensor method was subjected to an evaluation study using over 300 routine poultry samples and the results were compared with a microbial inhibition test. The cell-biosensor assay yielded many more suspect samples, 10.2% versus 2% with the inhibition test, which all could be confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Only one sample contained a concentration above the maximum residue limit (MRL) of 100 microg kg(-1), while residue levels in most of the suspect samples were very low (<10 microg kg(-1)). The method appeared to be specific and robust. Using an experimental set-up comprising the analysis of a series of three sample dilutions allowed an appropriate cut-off for confirmatory analysis, limiting the number of samples and requiring further analysis to a minimum.

Bioactivity-based screening of antibiotics and hormones.

Bioactivity-based screening methods are relatively cheap, quick and easy to use tools. Especially with respect to antimicrobial residues and compounds with hormonal activity, they form a very cost-effective alternative to physical chemical methods in large-scale surveillance and monitoring programs, where their main purpose is to identify samples that require additional chemical confirmation. A major advantage is their intrinsic capability to detect unknown compounds and new hazards. This review shows an overview of the available methods and their potential and limitations for regulatory control.

Improved microbial screening assay for the detection of quinolone residues in poultry and eggs.

An improved microbiological screening assay is reported for the detection of quinolone residues in poultry muscle and eggs. The method was validated using fortified tissue samples and is the first microbial assay to effectively detect enrofloxacin, difloxacin, danofloxacin, as well as flumequine and oxolinic acid, at or below their EU maximum residue limits (MRL). The accuracy of the assay was shown by analysing incurred tissue samples containing residue levels around the MRL. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) quantification of the quinolone concentration in these samples showed that the test plate can be used semi-quantitatively, allowing the definition of an "action level" as an inhibition zone above which a sample can be considered "suspect". The presented assay is a useful improvement or addition to existing screening systems.

Enigmatic Gratuitous Induction of the Covalent Flavoprotein Vanillyl-Alcohol Oxidase in Penicillium simplicissimum.

When Penicillium simplicissimum is grown on veratryl alcohol, anisyl alcohol, or 4-(methoxymethyl)phenol, an intracellular covalent flavin-containing vanillyl-alcohol oxidase is induced. The induction is highest (up to 5% of total protein) during the growth phase. In addition to vanillyl-alcohol oxidase, an intracellular catalase-peroxidase is induced. Induction of vanillyl-alcohol oxidase in P. simplicissimum is prevented by the addition of isoeugenol to veratryl alcohol-containing media, but growth is unaffected. The inhibitory effect of isoeugenol on induction is not observed when anisyl alcohol or 4-(methoxymethyl)phenol is used as the growth substrate. Based on the induction experiments and the degradation pathways for veratryl and anisyl alcohol, we propose that induction of vanillyl-alcohol oxidase is superfluous when P. simplicissimum is grown on these aromatic alcohols. However, the enzyme plays an essential role in the degradation of the methyl ether of p-cresol, 4-(methoxymethyl)phenol.

Crystallographic and kinetic evidence of a collision complex formed during halide import in haloalkane dehalogenase.

Haloalkane dehalogenase (DhlA) converts haloalkanes to their corresponding alcohols and halide ions. The rate-limiting step in the reaction of DhlA is the release of the halide ion. The kinetics of halide release have been analyzed by measuring halide binding with stopped-flow fluorescence experiments. At high halide concentrations, halide import occurs predominantly via the rapid formation of a weak initial collision complex, followed by transport of the ion to the active site. To obtain more insight in this collision complex, we determined the X-ray structure of DhlA in the presence of bromide and investigated the kinetics of mutants that were constructed on the basis of this structure. The X-ray structure revealed one bromide ion firmly bound in the active site and two bromide ions weakly bound on the surface of the enzyme. One of the weakly bound ions is close to Thr197 and Phe294, near the entrance of the earlier proposed tunnel for substrate import. Kinetic analysis of bromide import by the Thr197Ala and Phe294Ala mutants of DhlA at high halide concentration showed that the rate constants for halide binding no longer displayed a wild-type-like parabolic increase with increasing bromide concentrations. This is in agreement with an elimination or a decrease in affinity of the surface-located halide-binding site. Likewise, chloride binding kinetics of the mutants indicated significant differences with wild-type enzyme. The results indicate that Thr197 and Phe294 are involved in the formation of an initial collision complex for halide import in DhlA and provide experimental evidence for the role of the tunnel in substrate and product transport.