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GC-NPD and GC-MS analysis of preserved tissue of Bhopal gas disaster: evidence of methyl carbamylation in post-mortem blood.

Twenty-five preserved autopsy blood samples of Bhopal toxic gas exposed victims were analysed by gas chromatography (GC) coupled with either Nitrogen-Phosphorous detector (NPD) or mass spectrometer (MS) for the presence of methyl carbamyl valine in terms of valine methyl hydantoin (VMH). 84% of these samples showed a positive test for VMH on GC-NPD and the identity of the peaks were further confirmed on GC-MS. The concentration of VMH in the gas-affected positive blood samples ranged from 2.56 to 51.28 nanomoles. These results indicate entry of methyl isocyanate (MIC), one of the constituents of the toxic cloud caused by the disaster, into the blood stream of victims who had inhaled gas.

Accidents, Occupational↗

Histoenzymatic study of human renal tissue preservation: II--Catalase activity in proximal tubular cells is uncorrelated with transplant evolution.

Enzyme histochemical activity of catalase, a peroxisomal enzyme involved in cellular antioxidant systems, was studied in proximal tubular cells of human renal transplants as a marker of ischemia-reperfusion injury in the prediction of the evolution of renal transplants. A low enzymatic activity was observed in all renal biopsies performed at 30 min. reperfusion with no difference between the several evolution types of renal transplants. Reduced catalase activity due to ischemia-reperfusion injury could not be correlated with renal function or used as an index of renal function recovery.

Adenosine↗

Preserved tissue allografts in reconstructive surgery.

Results of treatment with three various kinds of allografts: lyophilized bone, deep frozen bone and cartilage preserved in physiological solution, all of them radiation-sterilized are presented. We believe that this presentation may be helpful in estimating the tissue bank's allografts and in establishing indications and contraindications in the application of allografts in surgery.The 'indices of coincidence' were compared in a group of 1014 patients after bone (lyophilized and radiation-sterilized) transplantation. It seems that such a variable as 'rebuilding of graft' may be of prognostic value in analysing the results of treatment in this group.The application of frozen and radiation-sterilized allogenic bone grafts for reconstructions is also described. An analysis of the results of treatment in 1125 patients reveals that the use of preserved bone reduces the extent and duration of surgery. Almost total substitution of grafts may be seen in 3-8 months after surgery.Allogenic, preserved cartilage is often used in facial reconstructions of face. Human costal cartilage, preserved in 0.9% NaCl and radiation-sterilized, was used for reconstruction. The patients were examined 24-190 months after surgery (in several clinical units) and results were collected in a special questionnaire by the team that performed surgery. In an analysed group of 437 patients after cartilage transplantation, 42.2% were operated because of posttraumatical changes, 29.0% because of congenital malformations and in 16.7% non-specific inflammations were the cause of reconstructive operations. Malformations were located mainly in the nose (59%), the ear concha (16.5%) and 10.9% were mandible.The results of treatment were compared with ages of patients, diagnosis and the locations of the changes. Very good results were achieved in 33.5% of the patients, and satisfactory in 41.8% of the patients. However, in 19.9% of the patients the result of treatment was unsatisfactory. Correlation between some clinical and biological characteristics and the result of treatment is under discussion.

Journal Article↗

Application of cryotechniques in cartilage tissue preservation and immunoelectron microscopy: potentials and problems.

Cryotechnical processing of cartilage has the potential to solve many of the tissue-specific problems associated with various routine chemical fixation protocols. This is particularly the case with respect to extracellular matrix architecture, the distortion or destruction of which (caused by extraction and/or precipitation of proteoglycan molecules) may be prevented. Adoption of such techniques also permits high-sensitivity immunoelectron-microscopy of the extracellular matrix space (carbohydrate epitopes). However, a number of difficulties still remain to be resolved, particularly that of matrix-cell interface separation occurring during freeze substitution and low temperature embedding. These problems are briefly addressed and possible solutions outlined.

Animals↗

A new system for cutting brain tissue preserving vessels: water jet cutting.

The water jet cutting system allows transaction and dissection of biological structures with little bleeding. Structures of higher tissue rigidity remain unchanged while softer tissues are mechanically dissected. In brain tissue, all vessels larger than 20 microns are left intact after the passage of the jet stream with a pressure of up to 5 bar, and therefore vessels can be isolated selectively from the surrounding tissue. Oedema is present adjacent to the cut and no increase of temperature occurs.

Animals↗

Antibacterial peptide PR-39 affects local nitric oxide and preserves tissue oxygenation in the liver during septic shock.

The effects of the antibacterial peptide PR-39 on nitric oxide (NO) and liver oxygenation (pO(2)) in a mouse model of endotoxaemia have been explored. In vivo electron paramagnetic resonance (EPR) spectroscopy was used to make direct measurements of liver NO and pO(2). Measurements of pO(2) were made at two different anatomical locations within hepatic tissue to assess effects on blood supply (hence oxygen supply) and lobule oxygenation; selectively from the liver sinusoids or an average pO(2) across the liver lobule. PR-39 induced elevated levels of liver NO at 6 h following injection of lipopolysaccharide (LPS) as a result of increased iNOS expression in liver, but had no effect on eNOS or circulatory NO metabolites. Sinusoidal oxygenation was preserved, and pO(2) across the hepatic tissue bed improved with PR-39 treatment. We propose that the beneficial effects of PR-39 on liver in this septic model were mediated by increased levels of local NO and preservation of oxygen supply to the liver sinusoids.

Animals↗

Ovarian tissue preservation and future fertility: emerging technologies and ethical considerations.

Both adult women and prepubertal girls facing cancer treatment may have the option to harvest and cryopreserve ovarian tissue. If they have successful therapy for their disease, the tissue may be autotransplanted back into the woman's body or follicles may be harvested and matured in vitro. These techniques all remain experimental, however, and should be performed only by specialized centers that can provide a multidisciplinary team. The procedures should be done under approval from an Institutional Review Board with proper assurance of informed consent. Although the costs for research could be assessed to patients if no research funds are available, clinical fees should not be charged at this stage in the development of these technologies. Consideration should be given to the protection of minors by ensuring parental informed consent and child assent whenever possible. Disposition of cryopreserved gonadal tissue in the event of the participant's death should be formally designated. A registry should be established to follow the health of participants and their eventual offspring.

Adolescent↗

Tissue preservation for single-tooth anterior esthetics.

Placing implants in the esthetic zone can be difficult because of the expectations of the patient. Inevitably, the hard and soft tissue architecture is lost after extraction in the traditional two-stage procedure, and additional surgical procedures are required to restore the tissue that has been lost. The authors describe techniques that can be applied to retain ideal soft tissue if it is present and predictably restore tissue that is less than ideal.

Adolescent↗

Regulation of fibrillin-1 by biglycan and decorin is important for tissue preservation in the kidney during pressure-induced injury.

There is growing evidence that the two small leucine-rich proteoglycans biglycan and decorin regulate the assembly of connective tissues and alter cell behavior during development and pathological processes. In this study, we have used an experimental animal model of unilateral ureteral ligation and mice deficient in either biglycan or decorin. We discovered that pressure-induced injury to the wild-type kidneys led to overexpression of decorin, biglycan, fibrillin-1, and fibrillin-2. In contrast, in biglycan-deficient kidneys the overexpression of fibrillin-1 was markedly attenuated and this was associated with cystic dilatation of Bowman's capsule and proximal tubules. Notably, we found that in ligated kidneys from decorin-null mice, fibrillin-1 expression was initially enhanced to the same extent as in wild-type animals. However, long-term obstruction resulted in down-regulation of fibrillin-1 and concurrent cystic dilatation of Bowman's capsule in 33% of kidneys at 5 months after obstruction. In all of the genotypes, no differences in fibrillin-2 expression were observed. These in vivo data correlated with a significant induction of fibrillin-1 expression in renal fibroblasts and mesangial cells by recombinant biglycan and decorin. Our results indicate a novel role for decorin and biglycan during pressure-induced renal injury by stimulating fibrillin-1 expression.

Animals↗

[Distal biceps tendon ruptures--experiences with soft tissue preserving reinsertion by bone anchors].

The repair of the distal biceps tendon rupture with bone anchors achieves excellent results. The important function of the M. biceps brachii as a powerful supinator and flexor can be reestablished and the soft tissue can be protected. We describe the repair of the rupture of distal biceps tendon by an anatomic refixation of the tendon to the tuberositas radii with bone anchors through a single anterior incision. In a four year period we treated eight patients with this technique. All patients were men with a mean age of 50 years. In each case the mechanism of injury was a single traumatic event with a large load on the flexed elbow. In average surgery was performed within five days and the patients stayed in the hospital for two days. Clinical follow up after almost two years showed no nerve damage or heterotopic bone formation. The range of motion and isometric muscle testing were almost symmetrical. All patients were able to return to preinjury activity level without any limitations.

Adult↗

[Soft tissue preservation and reconstruction in non-supporting foot parts].

The foot has two skin surfaces which differ on a morphological and functional basis. The plantar skin is a highly specialized organ which has a dash-pot-like effect in weight bearing. The skin which does not participate in weight bearing covers tendons and a very rich syndesmotic complex. The internal soft tissue is segmented in tiny compartments and therefore ist not extensible. The skin and the rather thin subcutaneous layer are not extensible either because of the proximity and the rigidity of the underlying structures. In consequence, trauma to the soft tissue of the foot may cause compartment syndromes and skin defects, or secondary unstable scars which impede function. Compartment syndromes must be recognized early and treated by immediate release. Fresh skin defects need early coverage when tendons and joints are exposed. There is a variety of local flaps which are evenly good for replacement of retracting scars.

Foot Injuries↗

The interest of the neurological tissue preservation for the investigation of sudden infant death syndrome.

Sudden infant death syndrome (SIDS) is the main cause of post-natal infant mortality in developed countries with an incidence rate ranging from 1.5 to 2:1000 live births. Brainstem dysfunction of circuits which control respiration and cardiovascular stability may be involved in SIDS. Combined neuropathological and biochemical studies could elucidate some of these aspects. The relationships of these deaths to medico-legal problems could explain the scarce availability of these tissues. The need for a central SIDS tissue bank is emphasized and a protocol for the collection of CNS tissue is proposed. A detailed neuropathological study using this protocol allowed us to recognize pathological features in the CNS of 9 SIDS cases. Some features were related to the maturation of CNS, and others to possible infections. Gliosis was a common finding in most cases. The present state of SIDS investigation and results are also discussed.

Brain↗

Prolonged storage of fixative for electron microscopy: effects on tissue preservation for diagnostic specimens.

Prolonged storage of Karnovsky's fixative, at both 4 and -20 degrees C, is possible in the diagnostic electron microscopy setting. Ultrastructural detail was not compromised in specimens processed in fixative that had been stored for 6 months. Evidence of smooth muscle and neuroendocrine differentiation was present in the form of actin filaments/dense bodies and neurosecretory granules, respectively. No difference in preservation was detected between specimens fixed in freshly prepared Karnovsky's fixative and fixative that had been stored at either 4 or -20 degrees C for up to 6 months. Thus, community hospitals can be provided with Karnovsky's fixative on a semiannual basis for surgical pathology specimens that may require electron microscopy for diagnosis.

Fixatives↗

Adenovirus-mediated VEGF(121) gene transfer stimulates angiogenesis in normoperfused skeletal muscle and preserves tissue perfusion after induction of ischemia.

BACKGROUND: Administration of angiogenic factors stimulates neovascularization in ischemic tissues. However, there is no evidence that angiogenesis can be induced in normoperfused skeletal muscles. We tested the hypothesis that adenovirus-mediated intramuscular (IM) gene transfer of the 121-amino-acid form of vascular endothelial growth factor (AdCMV.VEGF(121)) could stimulate neovascularization in nonischemic skeletal muscle and consequently attenuate the hemodynamic deficit secondary to surgically induced ischemia. METHODS AND RESULTS: Rabbits and rats received IM injections of AdCMV.VEGF(121), AdCMV.Null, or saline in the thigh, 4 weeks (rabbits) or 2 weeks (rats) before femoral artery removal in the injected limb. In unoperated rats, at the site of injection of AdCMV.VEGF(121), we found 96% and 29% increases in length density of arterioles and capillaries, respectively. Increased tissue perfusion (TP) to the ischemic limb in the AdCMV.VEGF(121) group was documented, as early as day 1 after surgery, by improved blood flow to the ischemic gastrocnemius muscle measured by radioactive microspheres (AdCMV.VEGF(121)=5.69+/-0.40, AdCMV.Null=2.97+/-0.50, and saline=2.78+/-0.43 mL x min(-1) x 100 g(-1), P<0.001), more angiographically recognizable collateral vessels (angioscore) (AdCMV. VEGF(121)=50.58+/-1.48, AdCMV.Null=29.08+/-4.22, saline=11.83+/-1.90, P<0.0001), and improvement of the bioenergetic reserve of the gastrocnemius muscle as assessed by (31)P NMR spectroscopy. Follow-up studies showed that superior TP to the ischemic limb in the AdCMV.VEGF(121) group persisted until it was equalized by spontaneous collateral vessel development in untreated animals. CONCLUSIONS: IM administration of AdCMV.VEGF(121) stimulates angiogenesis in normoperfused skeletal muscles, and the newly formed vessels preserve TP after induction of ischemia.

Adenoviridae↗