Use of an autologous leucocyte and platelet-rich fibrin patch on hard-to-heal DFUs: a pilot study.

OBJECTIVE: Leucopatch is a leukocyte and platelet-rich fibrin patch that provides concentrated blood cells and signal substances to the surface of an ulcer. It is produced by centrifugation of the patient's own venous blood. The aim of this pilot multicentre cohort study was to evaluate effects of the leucocyte patch in patients with hard-to-heal diabetic foot ulcers (DFUs). METHOD: Non-ischaemic Wagner grade 1 or 2 DFUs with a duration of more than 6 weeks and a maximal area of 10cm² were included. Patients with >40% ulcer area change during a two-week run-in period were excluded. The treatment was applied once a week for up to 19 treatments or until the foot ulcer was completely epithelialised. The primary endpoint was healing within 20 weeks. RESULTS: Of the 60 patients who gave consent 16 were excluded during run-in period, 44 patients initiated study treatment and 39 were included in the per-protocol analysis. Complete epithelisation was achieved in 34% (per-protocol analysis 36%) at 12 weeks and 52% (59%) at 20 weeks. In patients with ulcer duration less than 6 months, 73% of ulcers healed within 20 weeks. Patients with healed ulcers had larger ulcer area reduction during the first two treatment weeks compared to non-healers. Adverse events were mild and rare. CONCLUSION: The leucocyte patch is well-tolerated, easy to use and has potential in the armamentarium of the DFU treatment, provided this outcome is confirmed in an appropriately powered randomised clinical trial.

Successful heterologous expression of a novel chitinase identified by sequence analyses of the metagenome from a chitin-enriched soil sample.

Chitin and its derivative chitosan are abundant natural polysaccharides with many potential industrial applications. Metagenomic analysis of chitin-enriched soil samples using the Roche Genome Sequencer FLX platform led to the identification of several novel genes for chitin and chitosan modifying enzymes (CCMEs) which may be used to produce novel chitosans. The sequencing approach yielded 2,281,090 reads with an average length of 378 bp amounting to a total sequence information of approximately 851 Mb. Assembly of the obtained sequences comprised 699,710 reads representing 30.68% of all reads. A total of 6625 contigs larger than 500 bp containing 16,289 predicted genes are included in the assembly. Taxonomic profiling of the indigenous microbial community by applying the software CARMA revealed that 96.1% of the reads were of bacterial origin including 17% assigned to the family Xanthomonadaceae. Several putative genes encoding CCMEs were identified by comparison against the GenBank database, inclusive a full-length chitinase gene which was codon optimized for Escherichia coli and heterologously synthesized as a Strep-tagged protein in E. coli Rosetta 2 using the pET vector system. Approximately 5mg of the novel active chitinase was purified as demonstrated by dot assay analysis using glycol chitin as a substrate. Next generation metagenomic sequencing, thus, emerges as a new and powerful tool for the identification of potentially novel biocatalysts of biotechnological value.

Investigation of solid-state reactions using variable temperature X-ray powder diffractrometry. I. Aspartame hemihydrate.

PURPOSE: The object of this study was to demonstrate the applicability of variable temperature X-ray powder diffractometry (XRD) to investigate solid-state reactions using aspartame as a model compound. METHODS: Aspartame exists as a hemihydrate (ASH) under ambient conditions and converts to aspartame anhydrate (ASA) at approximately 130 degrees C. ASA on further heating to approximately 180 degrees C undergoes decomposition (intramolecular cyclization) to form a diketopiperazine derivative (DKP). The dehydration as well as the decomposition kinetics were studied isothermally at several temperatures. The unique feature of this technique is that it permits simultaneous quantification of the reactant as well as the product. RESULTS: While the dehydration of ASH appeared to follow first-order kinetics, the cyclization of ASA was a nucleation controlled process. The rate constants were obtained at various temperatures, which permitted the calculation of the activation energies of dehydration and cyclization from the Arrhenius plots. The activation energy of dehydration was also calculated according to the method described by Ng (Aust. J. Chem., 28:1169-1178, 1975) and the two values were in good agreement. CONCLUSIONS: The study demonstrates that XRD is an excellent complement to thermal analysis and provides direct information about the solid-states of various reaction phases.

Identifying male college students' perceived health needs, barriers to seeking help, and recommendations to help men adopt healthier lifestyles.

Seven focus groups at a university campus were formed to identify college men's health concerns, barriers to seeking help, and recommendations to help college men adopt healthier lifestyles. Content analysis was used to identify and organize primary patterns in the focus-group data. Results of the study revealed that the college men were aware that they had important health needs but took little action to address them. The participants identified both physical and emotional health concerns. Alcohol and substance abuse were rated as the most important issues for men. The greatest barrier to seeking services was the men's socialization to be independent and conceal vulnerability. The most frequently mentioned suggestions for helping men adopt healthier lifestyles were offering health classes, providing health information call-in service, and developing a men's center. Implications of the results are discussed.

[Clinical classification of diabetic foot as a guideline for selection of treatment]. / Kliniczne klasyfikacje owrzodzenia w zespole stopy cukrzycowej jako przewodnik w wyborze sposobu leczenia.

Foot ulceration in patients with diabetes mellitus is always a serious risk factor for lower limb amputation. To amputate or to continue conservative treatment is usually a very difficult decision and not without mistakes. The main objective of the study was assessment of the diabetic wound depth as a clinical guideline for decision-making of conservative or aggressive surgical treatment. Local foot ulceration and results of management of 62 diabetics were analysed. Lower leg amputations were performed in 40% of the patients. The foot wound was evaluated according to University of San Antonio or Wagner's scoring system. Conservative treatment resulted in failure in 19% of patients of grade I, 54% of grade II, 60% of grade III and 87% of grade IV according to Wagner's. Using the San Antonio classification there were 20% of amputations in grade I, 58% in grade II and 65% in grade III. Taking into account infection and peripheral vascular disease there were 37% of amputations in stage B, 41% in stage C and 80% in stage D. Data analysis of the study suggests better clinical usefulness of the San Antonio classification system in prognosis of limb salvage. There is high probability of failure in conservative treatment of diabetic foot lesion of grade II and III and in stage D according to University of San Antonio scoring system.

Hemostatic activity of organs in declamping shock.

The aim of the study was to determine the haemostatic components activity of organs in declamping shock. The abdominal aorta was cross-clamped below or above renal arteries. No significant changes in tromboplastic and antithrombin activities were found in the kidney, liver, lung, heart and skeletal muscle. Renal cortex and medulla as well as the lungs show higher plasminogen activator activity and considerably higher antiplasmin activity. Diminished fibrinolysis in the kidney and the lung may promote thrombotic complications.

Activity of membrane, cytosol and lysosome enzymes in organs and blood serum during declamping shock.

The aim of the study was to evaluate the enzyme activity of cellular membranes (GGT), cytosol (ALT, AST) and lysosome (AP, AcP) in the cytosol, whole homogenate and blood serum during declamping shock, following release of abdominal aorta cross-clamping. The aorta was clamped for 60 minutes. An increase in GGT, AP and AcP activities in the cytosol and whole homogenate of the renal cortex, renal medulla, liver, lung, heart and the skeletal muscle occurs after declamping. Rise in the enzymatic activity, especially of acid phosphatase is higher when the aorta above renal arteries was clamped. However, its activity in the blood serum remains unchanged. Alterations in the distribution and the activity of the studied enzymes may indicate that aortic clamping damages the endoplasmic reticulum and lysosomal membranes. Yet, cellular membranes preserve their structural and functional integrity.

An objective indicant of binocular vision in humans: size-specific interocular suppression of visual evoked potentials.

Evoked cortical potentials (VEPs) to grid patterns flashed to one eye were suppressed in amplitude when grid patterns were continuously presented to the other eye. The degree of interocular suppression of VEPs was influenced by the stereoacuity of the subjects. VEPs were obtained to each of two grid sizes flashed to one eye (individual squares subtending 15 and 60 min of arc) and changes in amplitude of these VEPs were considered as a function of four stimuli continuously presented to the other eye (diffuse light, 15, 30, and 60 min of arc squares in grids). Interocular suppression of VEPs was greater (a) when the continuously presented grid was of high (38.00 mL) as compared to low (00.38 mL) intensity, (b) when the continuous and flashed grids were of the same as compared to different sizes, and (c) in six subjects who had good as compared to six subjects who had poor binocularity. Eleven of the twleve subjects could be classified correctly as having good or poor binocularity on the basis of statistically significant interocular suppression of VEPs. The results were interpreted in terms of centrally located binocular neurons responsive to specific grid sizes or spatial frequencies and the decreased functioning of such neurons in subjects with poor binocularity.