Risk assessment of a cold argon plasma jet in respect to its mutagenicity.

Cold atmospheric pressure plasmas represent a favorable option for the treatment of heat sensitive materials and human or animal tissue. Beneficial effects have been documented in a variety of medical conditions, e.g., in the treatment of chronic wounds. It is assumed that the main mechanism of the plasma's efficacy is mediated by a stimulating dissipation of energy via radiation and/or chemical energy. Although no evidence on undesired side effects of a plasma treatment has yet been presented, skepticism toward the safety of the exposure to plasma is present. However, only little data regarding the mutagenic potential of this new treatment option is available. Accordingly, we investigated the mutagenic potential of an argon plasma jet (kinpen) using different testing systems in accordance with ISO norms and multiple cell lines: a HPRT1 mutation assay, a micronucleus formation assay, and a colony formation assay. Moderate plasma treatment up to 180 s did not increase genotoxicity in any assay or cell type investigated. We conclude that treatment with the argon plasma jet kinpen did not display a mutagenic potential under the test conditions applied and may from this perspective be regarded as safe for the use in biomedical applications.

Induction of proliferation of basal epidermal keratinocytes by cold atmospheric-pressure plasma.

BACKGROUND: Over the past few decades, new cold plasma sources have been developed that have the great advantage of operating at atmospheric pressure and at temperatures tolerable by biological material. New applications for these have emerged, especially in the field of dermatology. Recently it was demonstrated that cold atmospheric-pressure plasma positively influences healing of chronic wounds. The potential of cold plasma lies in its capacity to reduce bacterial load in the wound while at the same time stimulating skin cells and therefore promoting wound closure. In recent years, there have been great advances in the understanding of the molecular mechanisms triggered by cold plasma involving signalling pathways and gene regulation in cell culture. AIM: To investigate cold plasma-induced effects in ex vivo treated human skin biopsies. METHODS: Human skin tissue was exposed to cold plasma for different lengths of time, and analysed by immunofluorescence with respect to DNA damage, apoptosis, proliferation and differentiation markers. RESULTS: After cold plasma treatment, the epidermal integrity and keratin expression pattern remained unchanged. As expected, the results revealed an increase in apoptotic cells after 3 and 5 min of treatment. Strikingly, an induction of proliferating basal keratinocytes was detected after cold plasma exposure for 1 and 3 min. As these are the cells that regenerate the epidermis, this could indeed be beneficial for wound closure. CONCLUSION: We investigated the effect of cold plasma on human skin by detecting molecules for growth and apoptosis, and found that both processes are dependent on treatment time. Therefore, this approach offers promising results for further applications of cold plasma in clinical dermatology.

Senile hair graying: H2O2-mediated oxidative stress affects human hair color by blunting methionine sulfoxide repair.

Senile graying of human hair has been the subject of intense research since ancient times. Reactive oxygen species have been implicated in hair follicle melanocyte apoptosis and DNA damage. Here we show for the first time by FT-Raman spectroscopy in vivo that human gray/white scalp hair shafts accumulate hydrogen peroxide (H(2)O(2)) in millimolar concentrations. Moreover, we demonstrate almost absent catalase and methionine sulfoxide reductase A and B protein expression via immunofluorescence and Western blot in association with a functional loss of methionine sulfoxide (Met-S=O) repair in the entire gray hair follicle. Accordingly, Met-S=O formation of Met residues, including Met 374 in the active site of tyrosinase, the key enzyme in melanogenesis, limits enzyme functionality, as evidenced by FT-Raman spectroscopy, computer simulation, and enzyme kinetics, which leads to gradual loss of hair color. Notably, under in vitro conditions, Met oxidation can be prevented by L-methionine. In summary, our data feed the long-voiced, but insufficiently proven, concept of H(2)O(2)-induced oxidative damage in the entire human hair follicle, inclusive of the hair shaft, as a key element in senile hair graying, which does not exclusively affect follicle melanocytes. This new insight could open new strategies for intervention and reversal of the hair graying process.

In vivo and in vitro evidence for autocrine DCoH/HNF-1alpha transcription of albumin in the human epidermis.

The presence of albumin in the human epidermis has been reported more than a decade ago, but until now, it was assumed that this protein is synthesized in the liver and transported to the avascular skin. To our knowledge, transcription of albumin in the human epidermis was never considered. In this report, we present for the first time evidence for autocrine synthesis of albumin in the human epidermis in keratinocytes in situ and in vitro. Using double immunofluorescence labelling, we identified that albumin colocalized together with its transcription factor PCD/DCoH/HNF-1alpha in suprabasal keratinocytes in human full-thickness skin sections and in keratinocytes cultured in serum-free medium. Moreover, albumin and HNF-1alpha protein expression was confirmed by Western blotting in undifferentiated and differentiated keratinocytes as well as in human epidermal suction blister roof extracts. Reverse-transcriptase polymerase chain reaction analysis from human epidermal keratinocytes and epidermal suction blister roofs revealed the transcription of albumin. Using in vivo fluorescence excitation spectroscopy at the surface of human skin, we confirmed albumin as a major constituent yielding a lambda(max) at 295 nm, which was assigned to the single tryptophan 214 fluorophore in this protein. This in vivo result is in agreement with albumin concentrations of 10(-3) M, underlining the importance of this protein in epidermal homeostasis.

Molecular evidence that halo in Sutton's naevus is not vitiligo.

Both halo naevus and vitiligo are acquired leucodermas of unknown aetiology. To date a significant contribution of oxidative stress through accumulation of hydrogen peroxide (H2O2) has been documented in the pathomechanism of vitiligo but not in halo naevus. Both epidermal pterin-4a-carbinolamine dehydratase (PCD) and catalase are sensitive markers to follow H2O2 concentration-dependent deactivation of these proteins. In situ protein expression of PCD and catalase was examined in full-skin biopsies from skin phototype-matched controls (n=5), untreated and treated vitiligo patients (n=5) and patients with untreated halo naevus in association with vitiligo (n=3). Vitiligo was treated with pseudocatalase (PC-KUS) only. Catalase levels were determined in epidermal suction blister extracts using fast protein liquid chromatography (FPLC). In addition, epidermal H2O2 levels were followed in vivo by Fourier-transform Raman spectroscopy. The results of this study ruled out a contribution of H2O2 in the millimolar range in the depigmentation process of halo naevus as previously documented in vitiligo. Therefore, it can be concluded that both leucodermas exercise distinct concentration-dependent H2O2 signalling in their pathomechanisms.

Separation of free iodide from other I-species in human serum - Quantification in serum pools and individual samples.

A method for the determination of free iodide in human serum has been developed. Iodide from pooled serum samples has been separated from the organic matter by SEC, subsequently freeze-dried and analyzed by ion chromatography. Investigations for recovery and precision have been carried out and provided sufficient results. For quality assurance ICP-MS has been taken additionally as a total I-detector. The iodide results of ICP-MS agree well with IC values. Iodine containing SEC- fractions from iodide-spiked samples has shown no increased I-values except that in the iodide fractions, proving that there has been no iodide conversion into other I-species (and vice versa) during the whole procedure. Free iodide from two serum pools of different healthy persons has been determined as 2.25 and 2.43 microg I(-)/l, respectively. The values are related to total iodine levels determined by ICP-MS. For comparative reasons a table of individual iodine and iodide values is presented.

cDNA sequences of Schistosoma japonicum coding for two cathepsin B-like proteins and Sj32.

Sequences of three cDNAs of Schistosoma japonicum are presented. Two of them code for the antigens Sj32 (hemoglobinase) and cathepsin B. The third clone represents a mutant derivative of the cathepsin B gene. The DNA and amino acid sequences are compared to their homologues from S. mansoni. The mutant cathepsin B, in which the Cys29 in the active center is replaced by Ser, is discussed with respect to a possible function.

Changes in resting energy expenditure and body composition in anorexia nervosa patients during refeeding.

Accurate prediction of the energy level necessary to promote weight restoration in patients with anorexia nervosa would be clinically useful. Resting energy expenditure (REE), respiratory quotient, and body composition were measured in 10 nonmedicated women with anorexia nervosa during a vigorous refeeding protocol. REE was measured three times per week by open-circuit indirect calorimetry after an overnight fast. Subjects ranged in age from 19 to 38 years and weighed 39.9 +/- 4.3 kg (mean +/- standard deviation) at admission. The refeeding protocol was as follows: phase 1, 1,200 kcal/day for 1 week (baseline); phase 2, an increase of 300 kcal/day for 1 week; phase 3, 3,600 kcal/day until target weight was reached; phase 4, 1,800 to 2,800 kcal/day (stabilization). REE was 30.0 +/- 6.4, 33.5 +/- 6.7, 37.3 +/- 6.6 and 34.5 +/- 4.4 kcal/kg body weight during phases 1, 2, 3, and 4, respectively. The Harris-Benedict equation overestimated phase 1 24-hour REE by a mean of 14% and underestimated REE in phases 2, 3, and 4 by a mean of 8%, 24%, and 23%, respectively. Skinfold measurements revealed percent body fat to be 12 +/- 4% at admission and 19 +/- 5% at discharge, with a mean of 48% of the weight gained during refeeding attributable to increased body fat. These findings indicate that refeeding in anorexia nervosa is associated with increased REE, which cannot be explained by increased body mass, and that caloric requirements for weight restoration in patients with anorexia nervosa are best determined by monitoring individual response.

Mineral- and trace element concentrations in human breast milk, placenta, maternal blood, and the blood of the newborn.

The concentrations of the essential trace elements Cu, Fe, and Zn, and of the mineral elements Ca, K, Mg, and P during the perinatal period in human placenta and in the blood of the mother and the newborn (cord blood) were determined. Breast milk (colostrum and transitory milk) was also included to permit correlations between the different compartments. No correlations were found. The uptake by nutrition and the body-pools of the mother and their mobilization for these elements seem to be sufficiently high for an adequate supply of the fetus and the milk in the geographical region of Munich (Bavaria, FRG) under these investigations. Differences in the mineral-and trace element concentrations of colostrum and transitory milk for the elements P and Zn and to a lesser extent for Ca and Mg were observed. Additionally, breast milk samples from different geographical regions in Bavaria were investigated. Results for the heavy metals Cd, Hg, and Pb, and for the essential trace element Se are also presented for these samples, and can be seen as a reflection of the overall environmental and dietary influences during pregnancy in these geographical regions. ICP (Inductively Coupled Plasma)-emission spectrometry was used for the determination of the elements Cu, Fe, Zn, Ca, K, Mg, and P. For the additionally given elements in the milk-samples anodic stripping voltammetry (DPASV) (Cd, Pb), hydride atomic absorption spectrometry (AAS) (Se), and cold vapor AAS (Hg) were applied.

Selenium, cadmium, lead, and mercury concentrations in human breast milk, in placenta, maternal blood, and the blood of the newborn.

The concentrations of the trace elements Cd, Hg, Pb, and Se during the perinatal period in human placenta and in the blood of the mother and the newborn (cord blood) were determined. Breast milk (colostrum and mature milk) was also included to permit correlations between the different compartments. For Cd, a placental barrier exists, in accord with previous observations. For Pb, a strong correlation between the concentrations in the blood of the mother and of the newborn was found. The concentration of Hg was in most cases below the detection limit. Its concentration in colostrum was higher than in the mature milk. The results for Se reflect the knowledge about an essential trace element. Strong positive correlations were noted between maternal blood and cord blood and maternal milk. Anodic stripping voltammetry (DPASV) was used for the determination of Cd and Pb, cold vapor AAS (CVAAS) for the determination of Hg, and instrumental neutron activation analysis (INAA) for the determination of Se.

[Mineral and trace elements in human urine]. / Mineral- und Spurenelemente im menschlichen Urin.

Mineral and trace elements (Al, B, Ca, Cu, Fe, K, Li, Mg, Na, P and Zn) in 24 hour human urine samples from approximately 100 healthy persons (age between 20 and 50 years) were analysed by ICP (Inductively Coupled Plasma) and DCP (Direct Current Plasma) emission spectroscopy. The obtained values can be taken as "reference values" owing to the large number of individual samples. Mean values and the range of substance concentrations and substance excretion per 24 hours are given. Physiological reference values are reported for Ba, Sr and Ti, which are of particular interest in the field of occupational medicine; examples of increases in these three elements as a result of exposure at the work place are also given. Additionally some average values for Cd, Co, Ni and Pb have been measured by a voltammetric technique.

[Effect of an oxypolygelatine solution (Gelifundol) on the blood viscosity of healthy subjects]. / Einfluss einer Oxypolygelatinelösung (Gelifundol) auf die Fliesseigenschaften des Blutes gesunder Probanden.

Infusion of 500 ml oxypolygelatin (Gelifundol, 5.5% solution) in eight healthy volunteers led to hemodilution wiht consecutive changes of rheological parameters. Blood viscosity was statistically significantly lowered whereas plasma viscosity values showed a slight increase. RBC aggregation was slightly increased, RBC deformability remained unchanged.

[Improvement of blood viscosity after infusion of low-molecular hydroxyethyl starch (Expafusin) in healthy subjects]. / Verbesserung der Fliesseigenschaften des Blutes nach Infusion von niedermolekularer Hydroxyäthylstärke (Expafusin) bei gesunden Probanden.

In healthy volunteers, 500 ml of a 6% low molecular weight hydroxyethyl starch solution (Expafusin) was infused intravenously within 30 minutes. Blood samples for the measurement of rheological parameters were obtained before and after the infusion. Blood and plasma viscosity were significantly decreased after the infusion of HES. Red cell deformability as measured by a filtration technique was slightly improved. Red cell aggregation as measured by a light transmission method was markedly reduced. Repeated infusion of 500 ml Expafusin indicates that the improvement of the flow properties of blood can be maintained for more than 3 hours.

Health centre x-ray unit.

An analysis of the types and numbers of x-ray films requested in the first year of a health centre x-ray unit showed that chest films represented the largest proportion of these. The unit is most valuable when it is inmediately available to the patient and general practitioner at the time of consultation, and thus it should be open for at least five sessions per week. The likely referral rate for the health centre x-ray unit is 84 patients per 1,000 at risk, and a unit functioning for five sessions a week can examine 60 patients during that time. This minimum of five sessions would be fully used by a population of 30,000 patients. The running costs were found to be about the same as those of a hospital x-ray unit.