Behavioural, morphological and electrophysiological assessment of the effects of type 2 diabetes mellitus on large and small nerve fibres in Zucker diabetic fatty, Zucker lean and Wistar rats.

BACKGROUND: Peripheral neuropathy is a common complication in type 2 diabetes mellitus (T2DM). The most common presentation is in the form of a distal axonal sensory-motor polyneuropathy that involves large and small nerve fibres in variable proportion. METHODS: Zucker Diabetic Fatty (ZDF), Zucker Lean (ZL) and Wistar Han (WH) rats were used to assess the behavioural, morphological and electrophysiological effects that T2DM have on peripheral large and small nerve fibres of 6- to 40-week-old rats. RESULTS: ZDF rats presented mechanical hypersensitivity that initially worsened in parallel to the progression of diabetes and eventually reverted at later stages of the disease. The reversal from hypersensitivity to hyposensitivity paralleled a reduction in the number of intraepithelial skin nerve terminals and in the nerve fibre lengths. However, no increased levels of degeneration of dorsal root ganglion neurons were observed. Nerve conduction studies showed a reduction in sensory and motor nerve conduction velocity (CV) in hyperglycaemic ZDF rats. Microneurography showed significant alterations in several parameters of activity-dependent slowing (ADS) of mechano-insensitive C-nociceptors in ZDF rats. Surprisingly, some of these changes were also observed in ZL rats. Moreover, we found spontaneous activity in all three strains implying that C-nociceptors become hyperexcitable and spontaneously active not only in ageing hyperglycaemic ZDF rats but also in age-matched and apparently normoglycaemic ZL and WH rats fed with the same diet. CONCLUSIONS: ZDF rats presented a diabetic neuropathy involving large and small nerve fibres; additionally, ZL and WH rats also showed early small abnormalities in C-fibres, clearly detected by microneurography SIGNIFICANCE: This study provides a functional description of large and small nerve fibre function in a diabetic model that recapitulates many of the findings observed in patients suffering from type 2 diabetes mellitus.

Differential MMP-9 activity in CD34⁺progenitor cell-derived foam cells from diabetic and normoglycemic patients.

BACKGROUND: Upon coincubation with platelet aggregates, CD34(+) progenitor cells have the potential to differentiate into foam cells. There is evidence that progenitor cells from diabetic and nondiabetic patients have different properties, which may affect the patients' prognosis. In this study we investigated an in vitro model of foam cell formation based on patient-derived CD34(+) progenitor cells. We analyzed the growth characteristics as well as the M-CSF-release and matrix metalloproteinase (MMP) synthesis from CD34(+) progenitor cell-derived foam cells originating from diabetic and nondiabetic patients. METHODS AND RESULTS: Bone marrow samples were obtained from 38 patients who were elected for thoracic surgery. CD34(+) progenitor cells from diabetic and nondiabetic patients were isolated and incubated with platelets from healthy volunteers. Foam cell formation was confirmed by immunostaining (CD68) and quantified by light microscopy. Whereas the absolute number of foam cells was not affected, the negative slope in the growth curve was seen significantly later in the diabetic group. In supernatants derived from"diabetic" CD34(+) progenitor cells, MMP-9 was significantly enhanced, whereas MMP-2 activity or M-CSF-release was not affected significantly. CONCLUSION: In a coculture model of CD34(+) progenitor cells with platelets, we show for the first time that"diabetic" CD34(+) progenitor cells exhibit functional differences in their differentiation to foam cells concerning growth characteristics and release of MMP-9.

The recombinant bifunctional protein αCD133-GPVI promotes repair of the infarcted myocardium in mice.

BACKGROUND: Bone-marrow-derived progenitor cells are important in myocardial repair mechanisms following prolonged ischemia. Cell-based therapy of diseased myocardium is limited by a low level of tissue engraftment. OBJECTIVES: The aim of this study was the development of the bifunctional protein αCD133-glycoprotein (GP)VI as an effective treatment for supporting vascular and myocardial repair mechanisms. RESULTS: We have generated and characterized a bifunctional molecule (αCD133-GPVI) that binds both to the subendothelium of the injured microvasculature and to CD133(+) progenitor cells with high affinity. αCD133-GPVI enhances progenitor cell adhesion to extracellular matrix proteins and differentiation into mature endothelial cells. In vivo studies showed that αCD133-GPVI favors adhesion of circulating progenitor cells to the injured vessel wall (intravital microscopy). Also, treatment of mice undergoing experimental myocardial infarction with αCD133-GPVI-labeled progenitor cells reduces infarction size and preserves myocardial function. CONCLUSIONS: The bifunctional trapping protein αCD133-GPVI represents a novel and promising therapeutic option for limiting heart failure of the ischemic myocardium.

[Novel software-based and validated evaluation method for objective quantification of bone regeneration in experimental bone defects]. / Eine neue objektivierte und validierte Methode zur Quantifizierung der Knochenneubildung in Tiermodellen auf Basis eines frei verfügbaren Bildbearbeitungsprogramms.

AIM: The quantification of newly formed bone in experimental defect models is a problem in various experimental set-ups. Several methods have been described to evaluate and quantify the regeneration of newly formed bone in various animal models. Most methods only describe the amount of regenerated tissue on a semi-quantitative level, the results significantly depend on the subjective rating of the observer and such evaluation methods have not been validated in terms of objectivity and reliability. The aim of the present study was to introduce a novel evaluation method for the accurate quantification of bone regeneration on digital X-ray images using a freely available digital image software analysis programme (GIMP, GNU General Public Licence). METHODS: The method introduced here contains 5 steps: standardisation of size and colour, determination of range of interest (ROI), defining different qualities of mineralisation, pixel analysis with histogram function, similar to the Hondsfield index, and quantification. In order to evaluate the objectivity and reliability, the quantification method was compared to semi-quantitative scores described by Mosheiff and Werntz for inter- and intraobserver variability. Six observers were asked to determine bone regeneration in 16 X-ray images of 2 different animal models. In order to describe intraobserver variability, the evaluation was repeated after a period of 4 weeks. Statistical analysis including determination of intra- and interobserver variability (Bland-Altman coefficient of reproduction) was performed using SAS software. RESULTS: For both experimental set-ups analysed in this project (rabbit and sheep bone defects), the objectivity was significantly higher in the GIMP-based evaluation compared to the evaluation according to Mosheiff and Werntz using the Bland-Altman coefficient (rabbit: GIMP: 0.095, Mosheiff: 0.272, Werntz: 0.283; sheep: GIMP: 0.098, Mosheiff: 0.658, Werntz: 0.668). Analogous results were obtained for reliability (rabbit: GIMP: 0.086, Mosheiff: 0.221, Werntz: 0.385; sheep: GIMP: 0.102, Mosheiff: 0.339, Werntz: 0.623). CONCLUSION: This quantification method introduced here has proved to be a reliable and "easy-to-use" tool in order to perform objective quantification of bone regeneration in 2 different experimental set-ups. It offers a more detailed and quantitative way for precise determination of regenerated tissue and is characterised by higher objectivity and reliability compared to other semi-quantitative evaluation methods. The objectivity seems to be independent of the animal model to which the method is applied.

Orbital photogalvanic effects in quantum-confined structures.

We report on the circular and linear photogalvanic effects caused by free-carrier absorption of terahertz radiation in electron channels on (001)-oriented and miscut silicon surfaces. The photocurrent behaviour upon variation of the radiation polarization state, wavelength, gate voltage, and temperature is studied. We present the microscopic and phenomenological theory of the photogalvanic effects, which describes well the experimental results. In particular, it is demonstrated that the circular (photon-helicity sensitive) photocurrent in silicon-based structures is of pure orbital nature originating from the quantum interference of different pathways contributing to the absorption of monochromatic radiation.

Operative treatment of acute Achilles tendon rupture: Open end-to-end-reconstruction versus reconstruction with Mitek-anchors.

PURPOSE: To retrospectively compare the open end-to-end repair versus repair using the Mitek-anchor system in acute Achilles tendon rupture. METHOD: Forty-seven consecutive patients with Achilles tendon rupture, all operated on between 2004 and 2005, were included. Their medical records were reviewed and they were interviewed for surveillance of post-operative function at follow-up. Functional outcome was determined using an adapted VISA tendinopathy questionnaire and by testing the isometric ankle plantar flexion strength. Post-operative complications and recurrence rate of rupture were noted. RESULTS: Seven patients were lost to follow-up. From a total of 40 patients, twenty-eight (68% of total) underwent classic repair and 12 (32%) were treated by the Mitek-anchor system. Median age was 43 years (range 29-63). Median post-operative follow-up was 29 months (range 17-40). Median time to resume work was nine weeks in the classic group versus 12 weeks in the Mitek-group. Median time to resume sports was 19 versus 31 weeks respectively. Wound infections occurred in five percent of the total (one in each group) and rupture recurrence rate was three percent of the total (nil in classic group, one in Mitek-group). Loss of strength in the injured leg compared to the non-injured leg was greater in the Mitek-group than in the classic group. CONCLUSION: We conclude that in comparing classical end-to-end repair of acute Achilles tendon ruptures with repair using Mitek-anchors, it took patients in the Mitek-group longer to return to work and sport activities than in the classic group. Greater loss of strength in the injured leg was seen in the Mitek-group. Therefore we do not advocate the use of Mitek-anchors for the repair of acute ruptured Achilles tendons.