The influence of nitric oxide synthase 2 on cutaneous wound angiogenesis.

BACKGROUND: Inducible nitric oxide synthase (nitric oxide synthase 2, NOS 2) inhibition significantly suppresses chronically ischaemic skin flap survival, possibly because of reduced angiogenesis. OBJECTIVES: To investigate the effect of genetic NOS 2 inhibition on cutaneous wound angiogenesis in two in vivo murine models. The impact of NOS 2 manipulation on vascular endothelial growth factor (VEGF)-A stimulated and fibroblast growth factor (FGF)-2 stimulated angiogenesis was also investigated in the Matrigel(®) plug assay. METHODS: (i) Matrigel plugs/incisional wounds: two groups of NOS 2-/- mice and two groups of wild-type (WT) mice had bilateral Matrigel plugs containing 500 ng mL(-1) VEGF-A or 1000 ng mL(-1) FGF-2 injected subcutaneously in the abdomen. A 2·5 cm long dorsal incisional skin wound was created and sutured closed in the same animals. Wounds and plugs were explored at 7 or 12 days. (ii) Excisional wounds: dorsal 0·5 × 1·0 cm excisional skin wounds were created in four groups (two NOS 2-/- and two WT) and explored at 7 or 14 days. Wounds and Matrigel plugs were examined histologically and morphometrically for determination of percentage vascular volume (PVV). RESULTS: The PVV in NOS 2-/- incisional wounds and excisional wounds was significantly less than in WT wounds (P = 0·05 and P < 0·001, respectively). The PVV was significantly less in VEGF-A stimulated Matrigel plugs compared with FGF-2 stimulated plugs in NOS 2-/- mice (P < 0·01), but not in WT mice. CONCLUSIONS: NOS 2 is significantly involved in angiogenic signalling in healing skin wounds, particularly within the first 7 days. However, Matrigel plug vascularization suggests that the role of NOS 2 in angiogenesis is related to VEGF-A but not FGF-2 stimulated angiogenesis.

Use and abuse of faecal occult blood tests in an acute hospital inpatient setting.

BACKGROUND/AIMS: The role of the faecal occult blood test (FOBT) is untested. The aims of this study were to define the use of FOBT in a general hospital setting and to determine its influence on patient management. METHODS: Case notes and laboratory reports were retrospectively reviewed in all FOBTs performed in 2006 across three acute hospitals, with specific reference to clinical setting, indication, influence over clinical decision-making and management. Both guaiac and immunological tests were performed on all specimens. RESULTS: A total of 330 patients aged 2-104 (mean 74) years, 47% men, had 461 tests performed. A positive result was recorded in one or both tests in 64% of patients. Evidence of dietary restriction was found in only eight (2%) of patients and 218 (66%) patients took one or more medications that could have caused a false positive result. Indications were mostly for overt or suspected gastrointestinal blood loss with or without anaemia and/or iron deficiency, but 5% were for non-bloody diarrhoea and 3% screening for colorectal cancer. Patient care was adversely affected or delayed in 54 patients (16%), mostly because of the result being the stimulus for the decision to refer or not for endoscopy. Only one was considered appropriate as a screening test for colorectal cancer. CONCLUSIONS: The FOBT was applied in clinically inappropriate settings without consideration to confounding issues, and often led to inappropriate clinical decisions with considerable cost to hospital and patient. There is no place for FOBT in an acute hospital setting.

Hydrogen peroxide modulates angiotensin II-induced contraction of mesenteric arteries from streptozotocin-induced diabetic rats.

Hydrogen peroxide (H(2)O(2)) contributes in the regulation of vascular tone, especially in pathological states. The role of H(2)O(2) and superoxide anion free radicals in angiotensin II (Ang II)-induced contraction of diabetic tissues was examined with the aim of elucidating the underlying mechanisms. Isometric tension in response to various drug treatments was measured in isolated superior mesenteric arteries of streptozotocin (STZ)-induced diabetic WKY rats using the Mulvany wire myograph. Compared to the normal (euglycaemic) arteries, the Ang II-induced contraction was significantly reduced in diabetic arteries. Superoxide dismutase (SOD; converts superoxide to H(2)O(2)) significantly reduced the contraction in both types of arteries -- an effect abolished by catalase (H(2)O(2) scavenger), suggesting that the SOD effect was mediated by H(2)O(2). Treatment with catalase had no effect on the Ang II contraction in euglycaemic arteries, but it raised the contraction in diabetic arteries to euglycaemic levels. This increase was similar to that observed with diabetic arteries incubated with L-NAME. Combined catalase and L-NAME treatment further enhanced the contraction in diabetic arteries, suggesting that the catalase effect was not mediated by nitric oxide (NO). The catalase effect was abolished by indomethacin treatment. These results suggest that attenuation of Ang II-induced contraction in diabetic tissues is modulated by endogenous H(2)O(2), the scavenging of which unmasks an indomethacin-sensitive (and therefore cyclooxygenase product-mediated) Ang II-induced contraction.

Increased cutaneous blood flow but impaired post-ischemic response of nutritional flow in obese children.

It is well known that cardiovascular risk factors and dyslipidemia along with obesity can be identified early in life. Serum lipids and lipoproteins influence skin vessel reactivity in adults. The purpose of this study was to investigate the cutaneous microcirculation and its correlation to dyslipidemia in obese children. Thirty-four obese children and an equal number of age- and sex-matched healthy children were enrolled in this study. Laser Doppler flowmetry and dynamic capillaroscopy were performed to measure total cutaneous blood flow and nutritional blood flow, respectively. A significantly higher cutaneous baseline flow and peak flow after a 1-min arterial occlusion was noticed in obese group as compared to normal control. Results of dynamic capillaroscopy revealed values of resting capillary blood cell velocity that did not differ from controls but a significantly lower peak CBV after a 1-min arterial occlusion in obesity. Plasma lipids examination showed a high triglycerides and low high density lipoprotein cholesterol levels in obesity. Pearson analysis detected no significant correlations between pCBV, baseline flow and peak flow and dyslipidemia in obesity. These results may be explained by the fact that the duration of dyslipidemia in our study population was not long enough to cause vascular damage or the dyslipidemia was not pronounced enough to have an impact on skin vessel activity tests. It is also possible, that the combination of methods used in the present study was not sensitive enough to detect an existing correlations between flow values and high triglycerides/low HDL-cholesterol values in the obese group. Our results suggest that all the responses of the cutaneous microcirculation were mainly due to physiological compensatory rather than pathological reactions in young obese children. The results of cutaneous microcirculatory assessment in obese children provide follow-up parameters for assessing the risk of cardiovascular diseases later in life.

Changes in relative light fluence measured during laser heating: implications for optical monitoring and modelling of interstitial laser photocoagulation.

Dynamic changes in internal light fluence were measured during interstitial laser heating of tissue phantoms and ex vivo bovine liver. In albumen phantoms, the results demonstrate an unexpected rise in optical power transmitted approximately I cm away from the source during laser exposure at low power (0.5-1 W), and a decrease at higher powers (1.5-2.5 W) due to coagulation and possibly charring. Similar trends were observed in liver tissue, with a rise in interstitial fluence observed during 0.5 W exposure and a drop in interstitial fluence seen at higher powers (1-1.5 W) due to tissue coagulation. At 1.5 W irradiation an additional, later decrease was also seen which was most likely due to tissue charring. Independent spectrophotometric studies in Naphthol Green dye indicate the rise in fluence observed in the heated albumen phantoms may have been primarily due to light exposure causing photobleaching of the absorbing chromophore. and not due to heat effects. Experiments in liver tissue demonstrated that the observed rise in fluence is dependent on the starting temperature of the tissue. Correlating changes in light fluence with key clinical endpoints/events such as the onset of tissue coagulation or charring may be useful for on-line monitoring and control of laser thermal therapy via interstitial fluence sensors.

Laser thermal therapy: utility of interstitial fluence monitoring for locating optical sensors.

Multipoint optical fluence measurements can potentially be used to detect coagulation-induced changes in optical propagation during interstitial laser thermal therapy. Estimating the dimensions of coagulation using on-line optical monitoring, which is applicable to treatments where the tip of the source fibre is not precharred, may be limited by the accuracy of the placement of optical sensors with respect to source fibres. A strategy has been developed to determine accurately the position of a four-sensor linear array, prior to treatment, using optical fluence data obtained from the sensors for low-power (< or = 0.5 W) irradiation. A minimum of four sensors in an array was required in order to develop a mathematical formulation for position determination that did not require tissue optical properties or laser power as input. Optical propagation was based on diffusion theory for homogeneous tissues in spherical geometry. Low input laser power is needed to ensure that there are no thermally induced changes in tissue optical properties not accounted for in the mathematical description. Experimental evaluation was performed in a tissue-equivalent liquid phantom using 0.5 W of 805 nm optical energy and a translatable isotropic optical sensor. For sensor locations with 2 mm spacing, placement accuracy of 0.67 mm was achieved. The accuracy improved to 0.13 mm as the sensor spacing increased to 5 mm.

Evaluation of codon optimized recombinant Plasmodium knowlesi merozoite surface protein-119 (pkMSP-119) expressed in Pichia pastoris.

Malaria causes high global mortality and morbidity annually. Plasmodium knowlesi has been recognised as the fifth human Plasmodium sp. and its infection is widely distributed in Southeast Asia. Merozoite surface protein-119 (MSP-119) appears as a potential candidate for malaria blood stage vaccine as it could induce protective immunity. In this study, codon optimized P. knowlesi MSP-119 (pkMSP-119) was expressed and purified in yeast Pichia pastoris expression system. The purified recombinant protein was further evaluated using Western blot assay using knowlesi malaria, non-knowlesi human malaria, non-malarial parasitic infections and healthy serum samples (n = 50). The sensitivity of purified pkMSP-119 towards detection of knowlesi infection was as 28.6% (2/7). pkMSP-119 did not react with all non-malarial parasitic infections and healthy donor sera, yet reacted with some non-knowlesi human malaria sera, therefore lead to a specificity of 86.0% (37/43).