Naturally occurring growth factors in porcine wounds treated with autologous keratinocytes in a liquid environment

A step toward an improved understanding of the complex mechanisms of growth factor interactions may lie in the detection of endogenous growth factors during normal wound healing. The findings of this study on standardized full thickness wounds in swine, provide direct evidence that growth factors were present in the wound fluid in the picogram range (highest concentrations ranging from 1273 pg/ml for transforming growth factor-beta (TGFβ) to 85.6 pg/ml for platelet-derived growth factor-AB (PDGF-AB ) during healing. The presence of transplanted autologous keratinocyte suspensions and cultured epithelial sheet graft had no significant effect upon the observed growth factor levels, although transplanted keratinocyte cell suspensions (KCS) and cultured epidermal autografts (CEA) did accelerate healing in comparison to control wounds in our model (KCS treated wounds healed in 13.2±0.9 days, CEA in 13.7 days±0.8 and control wounds in 14.7 days±0.3). The variable occurrence of growth factors during normal wound healing may suggest possible mechanisms of growth factor interaction which could have an impact on the future design of their therapeutic use. Received: 28 September 1998 / Accepted: 16 November 1998     

Epidemiology of pneumococcal infections in Swedish children

Objective: This paper provides an overview of pneumococcal infections in Swedish children.
Method: Data supplied by the Swedish Institute for Infectious Disease Control (SMI) provided information on invasive pneumococcal isolates and on isolates with reduced susceptibility to penicillin. Disease burden was estimated from data collected in northern Stockholm and Malmöhus County. Results: Only 3–6% of the total number of invasive pneumococcal isolates came from children 0–15 years of age. Predominant serotypes in descending frequency were 7, 6, 14, and 23. Strains from all sources with reduced sensitivity to penicillin (MIC 0.5 mg/l) were found in 3% of children and varied between 0.2% and 11%, with the highest value found in Southern Sweden (predominating strains were 9, 19, 15, 6, and 23). A 10-year review of all cases of meningitis in Northern Stockholm reflected an incidence of 10/100000 (0–2 years) or 5.8/100000 (0–5 years), with severe sequelae occurring in 20% of children. This information can be used to predict an annual incidence of 30 cases of meningitis in Sweden.
Conclusion: The large proportion of serotype 7 among invasive isolates is distressing since this serotype is not represented in the present 7- and 9-valent protein-conjugated vaccines under development. However, the heptavalent vaccine, including serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F would (at a serotype level) provide coverage against 83% of the resistant isolates in Southern Sweden.     

Wheezing following lower respiratory tract infections with respiratory syncytial virus and influenza A in infancy

We examined the incidence of subsequent wheezing in 292 children, hospitalized for influenza A or respiratory syncytial virus (RSV) lower respiratory tract infection, during two consecutive seasons (November–December, 1993 and March–April, 1995). Questionnaires concerning episodes of wheezing and known risk factors for wheezy bronchitis were mailed to parents 1 year after hospitalization. Sixty per cent of parents reported two or more episodes of wheezing following either influenza A or RSV. Hospitalization as a result of wheezing was necessary in 15% of the patients. The severity of the primary infection, as indicated by the need for treatment in the intensive care unit (ICU), was correlated with later wheezing. No additional significant risk factors predicting later wheezing could be identified.     

Receptor-activated Ca2+ increases in vibrodissociated cortical astrocytes: A nonenzymatic method for acute isolation of astrocytes

A new nonenzymatic method for the acute isolation of astrocytes from rat cerebral cortex is described. A vibratory device was used to dissociate the cells from thin brain slices, and the method yielded fresh and relatively well-preserved astrocytes without previous enzyme incubation. These cells were examined in a microspectrofluorometric system for measurement of changes in intracellular free calcium concentrations ([Ca²⁺]_i), and their expression of various neurotransmitter receptors was determined. Acutely isolated glial fibrillary acidic protein (GFAP)-positive astrocytes (p7–p18) were seen to respond to the metabotropic glutamate receptor agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD, 10^-4 M) with increases in [Ca²⁺]_i, and this response was blocked by (RS)-1-aminoindan-1,5 dicarboxylic acid (AIDA, 10^-3 M), an antagonist to group 1 metabotropic glutamate receptors. The δ-opioid receptor agonist D-Pen², D-Pen⁵-enkehalin (DPDPE, 10^-6 M) evoked [Ca²⁺]_i increases that were blocked by the δ-opioid antagonist ICI 174.388 (10^-5 M). The astrocytes failed to respond to 5-hydroxytryptamine (5-HT, 10^-5 M), although the same cells subsequently were found to respond to other agonists. Furthermore, [Ca²⁺]_i responses evoked by phenylephrine (10^-5 M) were blocked by prazosin (0.2⋅10^-6 M), suggesting the expression of α₁-adrenergic receptors on the acutely isolated astrocytes. The cells were also shown to react with [Ca²⁺]_i increases in response to depolarization with high extracellular potassium concentrations (50⋅10^-3 M). The signals induced by depolarization were not seen in Ca²⁺-free buffer, indicating the presence of voltage-activated calcium channels in these cells. Thus, the present study confirms some of the results earlier obtained in cell cultures, suggesting that cortical astrocytes in vivo express glutamate, opiate, and adrenergic receptors, coupled to increases in [Ca²⁺]_i, whereas no receptors for 5-HT could be detected. J. Neurosci. Res. 54:390–401, 1998. © 1998 Wiley-Liss, Inc.     

Immunoisolating encapsulation of intrathecally implanted bovine chromaffin cells prolongs their survival and produces anti-allodynic effect in spinally injured rats

We have previously reported that intrathecal (i.t.) implantation of bovine chromaffin cells has an anti-allodynic effect in a rat model of mechanical and cold allodynia-like neuropathic pain after spinal cord injury. The technique of encapsulation of the cells by a semipermeable membrane has been developed recently. The present study was undertaken to investigate the effects of encapsulated bovine chromaffin cells on the allodynia-like pain in the same model. Capsules with bovine chromaffin cells or control capsules were implanted in the spinal subarachnoidal space in rats. Their response in behavioural tests were recorded for 2 months. At termination, the capsules were explanted and examined morphologically with tyrosine hydroxylase immunohistochemistry. The mechanical allodynia was totally abolished from week 2 after implantation of the cells and throughout the 8-week test period. The abnormal cold response was also attenuated in about half of the animals. The threshold to acute nociceptive stimulation was not affected. Eight weeks after implantation, 60–80% of the encapsulated chromaffin cells were still tyrosine hydroxylase positive. No effects were observed with control capsules. The results indicate that spinal implantation of encapsulated xenogeneic chromaffin cells may be useful in treating some refractory painful states associated with spinal cord injury. Immunoisolation of chromaffin cells by a semipermeable membrane may inhibit immunorejection, prolong the survival of the cells and enhance their anti-allodynic effect.     

Inositol 1,4,5-trisphosphate turnover enzymes--activities and subcellular distribution in hepatocarcinogenesis

The metabolism of inositol 1,4,5-trisphosphate and inositol 1,3,4,5- tetrakisphosphate in homogenates and sub-fractions from normal rat liver and premalignant liver nodules was investigated. The activities of 5-phosphatase, expressed as pmol converted substrate per minute and mg protein, were equal when using the two substrates, and did not differ between normal and nodular homogenates. Subcellular fractions were purified by sequential steps of differential centrifugation and density gradient fractionation procedures. The total phosphatase activity was found to be distributed between cytosol (15%) and membraneous fractions (75%), with most of the enzyme activity residing in the plasma membranes. A doubling of phosphatase specific activity was seen in the nodular low density membrane fraction, containing Golgi apparatus and endosomes, as compared with normal liver. Inositol 1,4,5- trisphosphate 3-kinase activity was found to be exclusively cytosolic. No difference in this enzyme was seen between the two tissue types studied. Vasopressin (0.2 or 2 microM) had no effect either on phosphatase or kinase activity. The compartmentalization of inositol polyphosphate 5-phosphatase activity presents a possible explanation of earlier findings that premalignant liver tissue was able to respond with inositol 1,4,5-trisphosphate, but not inositol 1,3,4,5- tetrakisphosphate formation after agonist stimulation.      

Patterns of dermal exposure to hazardous substances in European union workplaces

Workplace dermal exposure assessment is a complex task that aims to understand the dynamic interaction between the skin and the hazardous substances present in the surrounding environment. A European project known as RISKOFDERM gathered dermal exposure data in 85 workplaces (industrial and other types) in five countries in Europe. In order to optimize data collection and to develop a representative picture of dermal exposure, scenarios (tasks made up of a series of activities) were grouped together into dermal exposure operation units (DEOs). The allocation of scenarios to relevant DEOs was achieved on the basis of similarities of exposure routes, tasks and professional judgement. Sampling and quantification procedures were based on the approaches recommended by the OECD protocol. The laboratories involved in the analysis of the samples participated in quality assurance programmes. This exercise resulted in 419 body measurements and 437 measurements on hands expressed in terms of formulation (product) in use. Exposures for a given scenario varied by several orders of magnitude. The extent and patterns of exposure were found to be dependent on various exposure determinants, including inter- and intra-scenario variations. Hands were found to be the most contaminated parts of the body. Exposure patterns for liquid and solid contaminants were different. On the basis of the analysis of the data presented here, the averaged results (median and 95th percentile) for a given DEO unit should not be used as a representative measure of dermal exposure for all scenarios within that DEO without taking the exposure determinants into account. However, the data could be used to develop an exposure matrix (indicative exposure distributions) for different types of scenario and workplace, using determinants of exposure and a Bayesian approach to integrating expert opinion.