Research and comparative characteristic of the factors determining elasticity of the human skin

Changes in skin elasticity have been analyzed under different conditions (upon skin stretching, at different thickness of the skin, at different contents of collagen, intercellular and endocellular liquids, upon changes of venous pressure, and during contraction and relaxation of smooth muscles in skin vessels - vasomotions). Elasticity was defined by the acoustic method from the speed of diffusion of a superficial sheared acoustic wave in the skin, by the autoresonant method from the mechanical resonance frequency of skin, and from the vacuum pressure needed for skin site deformation of constant volume. It was shown that the major factors determining the elasticity of skin are it stretching, thickness, and the contents of collagen and liquids in it. The influence on elasticity of venous pressure and the contraction activity of smooth muscles in vessels is not essential. This suggests that the parameters of skin elasticity can be used as indicators of systemic and local lesions of the connective tissue.     

Histone complements of human tissues,carcinomas, and carcinoma-derived cell lines

Summary The pattern of subtypes of the nucleosomal histories and of historic Hl was investigated in human cells from adult and fetal lung and liver, from carcinoma tissues and from carcinoma-derived cell lines, with the object of comparing these patterns, and their relationship to cell growth rate, with those in cells of other species. The subtype pattern of the nucleosomal histories H2A and H3 shows a correlation with replication rate. In adult tissues, subtype H3-3 predominates over H3-2 and H3-1, and the subtype H2A-1 and H2A-2 are approximately equally abundant. In fetal tissues, lung carcinoma and cultured carcinoma-derived cell lines, the subtype H3-1 is predominant and H2A-1 is more abundant than H2A-2. The subtype pattern of H 1 also differs between normal and carcinoma cells, among different tissues, and in different cell lines derived from the same type of carcinoma. In particular, the relative level of H1° differs in several cell lines showing relatively high rates of replication, and in some cases represents more than 25% of the total H1, similar to the level in slowly replicating normal adult liver and lung tissues. The relative level of H1° does not therefore appear to be correlated in a simple manner with cell growth rate in these human cells.Abbreviations PhMeS0² phenylmethylsulfonyl fluoride - SDS sodium dodecyl sulphate     

Accurate determination of the structural elasticity of human hair by a small-scale bending test

This paper reports on a small-scale bending method for human hair. The test sample, which is elliptical in cross-section, is fixed to a hollow steel needle using resin to form a cantilever. A loading probe is used to subject this to a lateral load, where the load is applied parallel to either the long or short axis of the elliptical cross-section. From these tests, load-displacement relationships for the hair were obtained. From the experimental data and analysis, we found that the structural elasticity determined is independent of the direction of bending, and precise measurements of the structural elasticity of human hair with scattering of less than 5% were realized using this test scheme. Finally, changes in the structural elasticity of hair due to hair treatments were detected and the changes are discussed based on a theoretical model of the multi-layered structure.     

Purification and characterization of L-DOPA decarboxylase from human kidney

L-DOPA decarboxylase has been purified to homogeneity from post mortem removed human kidneys. Homogeneity was examined by polyacrylamide gel electrophoresis (PAGE) analysis both in the presence and absence of SDS. The enzyme has a molecular weight of 100,000 daltons estimated by gel filtration and 50,000 daltons determined after SDS-PAGE. Human L-DOPA decarboxylase therefore is a dimer. Polyclonal antibodies produced against human L-DOPA decarboxylase react with the 50,000 daltons enzyme subunit after immuno-blotting and also precipitates enzyme activity. Activity against L-DOPA is partially inhibited by 5-hydroxytryptophan (5-HTP). The effect of various cations on L-DOPA decarboxylase activity has also been tested.     

Immunological characterization of human acid phosphatase gene products

The immunological cross-reactivity of heterogeneous acid phosphatase isozymes from different human tissues has been studied using monospecific antisera prepared against four homogeneous acid phosphatases. The enzyme characterized as tartrate-inhibitable, prostatic acid phosphatase is also found to be present in leukocytes, kidney, spleen, and placenta. The tartrate-inhibitable (liver) lysosomal enzyme is also found in kidney, fibroblasts, brain, placenta, and spleen, but it is not detectable in erythrocytes and prostate. In several tissues, 10–20% of the tartrate-inhibitable enzyme is not precipitated by any of the antisera used; an exceptionally high amount (54%) of such an enzyme is present in human brain. Antiserum against a low molecular weight tartrate-resistant liver enzyme (14 kDa) does not cross-react with the erythrocyte enzyme. (10–20 kDa). All other tissues except placenta, prostate, and fibroblast cells show a cross-reactivity with the 14-kDa acid phosphatase antiserum. Thus, the low molecular weight human liver acid phosphatase is distinct from the erythrocyte enzyme, and there are also at least three different tartrate-inhibitable acid phosphatases in human tissues. Chromosomal assignments have been made for only two of the (at least) five acid phosphatases that are present in adult human tissues.This study was supported by DHHS Research Grant GM 27003 from the U.S. National Institute of General Medical Sciences and by Grant SFB-104 from the Deutsche Forschungsgemeinschaft.     

In vitro deregulation of markers characteristic of human prostate epithelial cells

We have screened primary cultures of human prostate for the expression of markers reported to be characteristic of specific cell lineages in vivo, in order to ascertain whether human prostate cells in vitro maintain and reflect their in vivo differentiated phenotypes and to evaluate the homogeneity of the populations of cells that can be derived from this tissue. Using single and dual stain immunofluorescent microscopy to analyse very early organoid and subsequently derived monolayer stage cultures, we have observed that expression of markers characteristic of human prostate epithelial cells in vivo is deregulated within 48h, indicating that dissociation of human prostate tissue and cultivation of prostate epithelial cells in culture can result in promiscuous expression of cell type specific markers of prostate epithelial cells. These observations have important implications for studies of cell lineage and differentiation of prostate cells in vitro.     

Cell contraction caused by microtubule disruption is accompanied by shape changes and an increased elasticity measured by scanning acoustic microscopy

The state of crosslinking of microfilaments and the state of myosin-driven contraction are the main determinants of the mechanical properties of the cell cortex underneath the membrane, which is significant for the mechanism of shaping cells. Therefore, any change in the contractile state of the actomyosin network would alter the mechanical properties and finally result in shape changes. The relationship of microtubules to the mechanical properties of cells is still obscure. The main problem arises because disruption of microtubules enhances acto-myosin-driven contraction. This reaction and its impact on cell shape and elasticity have been investigated in single XTH-2 cells. Microtubule disruption was induced by colcemid, a polymerization inhibitor. The reaction was biphasic: a change in cell shape from a fried egg shape to a convex surface topography was accompanied by an increase in elastic stiffness of the cytoplasm, measured as longitudinal sound velocity revealed by scanning acoustic microscope. Elasticity increases in the cell periphery and reaches its peak after 30 min. Subsequently while the cytoplasm retracts from the periphery, longitudinal sound velocity (elasticity) decreases. Simultaneously, a two- to threefold increase of F-actin and alignment of stress fibers from the cell center to cell-cell junctions in dense cultures are induced, supposedly a consequence of the increased tension.     

Radiological toxicity of some fish and meat tissues consumed in southwestern Nigeria

The activity concentrations of natural radionuclides in 30 fish tissues and 30 meat organs have been determined using high purity germanium (HPGe) detector. For the fish tissues, the mean activity concentrations of ²³⁸U and ²³²Th were highest in Catfish (Clarias gariepinus) with values of 5.31 ± 1.30 and 6.09 ± 0.91 Bq/kg, respectively. The lowest mean activity concentrations of ²³⁸U and ²³²Th were seen in Titus (Scomberomorus tritor) and Croaker (Micropogonias undulatus), Croaker with values of 1.51 ± 2.08 and 64.42 ± 6.33 Bq/kg, respectively. The mean activity concentration of ⁴⁰K was highest in Micropogonias undulatus and lowest in Tilapia (Oreohronis niloticus) with values of 64.42 ± 6.33 and 6.53 ± 0.98 Bq/kg. For the meat organs, the highest mean activity concentration of ²³⁸U, ²³²Th, and ⁴⁰K was highest in kidney, liver, and heart, respectively with values of 2.82 ± 0.47, 4.57 ± 0.69, and 52.07 ± 7.81 Bq/kg. Small intestine had the lowest mean activity concentrations of ²³⁸U and ²³²Th with values of 1.14 ± 0.16 and 0.89 ± 0.08 Bq/kg, respectively. Beef had the lowest mean activity concentration of ⁴⁰K with a value of 17.61 ± 2.14 Bq/kg.     

Formation of 8-nitroguanine in tobacco cigarette smokers and in tobacco smoke-exposed Wistar rats

Our previous studies have shown that 8-nitroguanine (8-NO(2)-G) could serve as a specific biomarker of DNA damage induced by gaseous nitrogen oxides (NO(x)) exposure. To evaluate the effect of tobacco cigarette smoking on the DNA damage in peripheral lymphocytes of cigarette smoke ones, we randomly collected and determined the level of 8-NO(2)-G in DNA extracted from peripheral lymphocyte of 15 each of light-smoking healthy volunteer (L-S, less than one pack per day), moderate-smoking healthy volunteers (M-S, one to two pack per day for 5-10 years), heavy-smoking healthy volunteers (H-S, over two packs per day for 10 years), lung cancer patients with heavy smoking (cancer H-S) and non-smoking healthy controls. Both of the mean level of the 8-NO(2)-G levels in peripheral lymphocyte (0.90+/-1.0, 1.23+/-1.14, 1.43+/-0.79, 3.62+/-1.38 ng per microg DNA) and serum nitrite (38.99+/-9.58, 46.70+/-9.38, 55.46+/-10.45, 70.1+/-18.54 microM) of L-S, M-S, H-S and cancer H-S groups were higher than that of non-smoking healthy controls (0.02+/-0.04 and 18.96+/-4.31 for 8-NO(2)-G level and serum nitrite, respectively). Furthermore, in animal experiment, a dose-dependent increase in 8-NO(2)-G was observed in rat lung and peripheral lymphocyte DNA of Wistar rats after tobacco cigarette smoke exposure twice a day, for 1 month. The level of 8-NO(2)-G is 0.17+/-0.41, 1.65+/-3.15, 23.50+/-20.75 and 37.58+/-17.55 ng per microg lung DNA for rat exposed with tobacco cigarette smoke from 0, 5, 10, 15 cigarettes per day, respectively. It was also found that count of peripheral lymphocytes and nitrite concentration in serum of rat increased after the tobacco smoke exposure. It is postulated that tobacco cigarette smoking could induce DNA damage (8-NO(2)-G formation) by exo- and endogenous NO(x).     

Success rate of primary human endothelial cell culture from umbilical cords is influenced by maternal and fetal factors and interval from delivery

Summary There is increasing interest in human umbilical cord vein as a source of endothelial cells. This paper shows that success in setting up cultures of human endothelial cells from umbilical cords depends not only on culture conditions, as so far proposed, but also on factors preceding the harvesting of the cells. In particular, the mother's smoking habit and the use of umbilical cord within 1 h of delivery have been shown to impair success of the culture. Age, parity, diabetes, and hypertension of the mother, type of delivery, and sex and weight of the newborn did not significantly influence the possibility of establishing successful endothelial cell culture. This work was supported by the Italian National Research Council, Grant 82.00151.04.     

Quantitation of Type I to type III collagen ratios in small samples of human tendon,blood vessels,and atherosclerotic plaque

A method is described whereby the ratio of the major interstitial collagens (Types I and III) can be measured in biopsy specimens of human tissue weighing as little as 25 mg. Marker peptides are solubilized from the tissue by digestion with cyanogen bromide. These peptides which are not known to be involved in collagen crosslinking are isolated and quantified by a combination of carboxymethyl-cellulose chromatography and polyacrylamide gel electrophoresis. The peptides used are α1(I)-CB7 and α1(III)-CB5. The use of the method is illustrated by analyzing the collagen type ratio in small specimens of tendon, aorta, and vena cava.     

A comparative map of chicken chromosome 24 and human chromosome 11

To improve the physical and comparative map of chicken chromosome 24 (GGA24; former linkage group E49C20W21) bacterial artificial chromosome (BAC) contigs were constructed around loci previously mapped on this chromosome by linkage analysis. The BAC clones were used for both sample sequencing and BAC end sequencing. Sequence tagged site (STS) markers derived from the BAC end sequences were used for chromosome walking. In total 191 BAC clones were isolated, covering almost 30% of GGA24, and 76 STS were developed (65 STS derived from BAC end sequences and 11 STS derived within genes). The partial sequences of the chicken BAC clones were compared with sequences present in the EMBL/GenBank databases, and revealed matches to 19 genes, expressed sequence tags (ESTs) and genomic clones located on human chromosome 11q22-q24 and mouse chromosome 9. Furthermore, 11 chicken orthologues of human genes located on HSA11q22-q24 were directly mapped within BAC contigs of GGA24. These results provide a better alignment of GGA24 with the corresponding regions in human and mouse and identify several intrachromosomal rearrangements between chicken and mammals.     

Genome-wide profiling of gene expression in 29 normal human tissues with a cDNA microarray.

We have performed a comprehensive analysis of the expression profiles in 25 adult and 4 fetal human tissues by means of a cDNA microarray consisting of 23,040 human genes. This study revealed a number of genes that were expressed specifically in each of those tissues. Among the 29 tissues examined, 4,080 genes were highly expressed (at least a five-fold expression ratio) in one or only a few tissues and 1,163 of those were expressed exclusively (more than a ten-fold higher expression ratio) in a particular tissue. Expression of some of the genes in the latter category was confirmed by northern analysis. A hierarchical clustering analysis of gene-expression profiles in nerve tissues (adult brain, fetal brain, and spinal cord), lymphoid tissues (bone marrow, thymus, spleen, and lymph node), muscle tissues (heart and skeletal muscle), or adipose tissues (mesenteric adipose and mammary gland) identified a set of genes that were commonly expressed among related tissues. These data should provide useful information for medical research, especially for efforts to identify tissue-specific molecules as potential targets of novel drugs to treat human diseases.     

The stoichiometric activation of human skin fibroblast pro-collagenase by factors present in human skin and rat uterus

Purified human skin fibroblast collagenase zymogen was used as a substrate for activators partially purified from the medium of cultured human skin and rat uterus. Analysis of the activated enzyme by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that these activators do not produce measurable changes in the molecular weight of the zymogen. Kinetic analysis indicated a noncatalytic mechanism of action for these activators, since zymogen activation was independent of incubation time, and dependent only upon the concentration of the activator fractions. These results are consistent with a Stoichiometric mechanism of procollagenase activation by these macromolecules.     

Determination of multielement concentrations in normal human organs from the Japanese

In order to elucidate the distribution of elements in organs from healthy Japanese, instrumental neutron activation analysis (INAA), based on the preliminary examination, was applied to quantitative determination of multielements in nine organs autopsied (brain, heart, kidney, liver, lung, muscle, pancreas, spleen, and thyroid). The following results were obtained: (1) The values obtained for each element could be considered to be representative as "normal values" and "ranges" in organs from healthy Japanese males; (2) the essential elements Br, Cl, Co, Cu, Fe, K, Mn, Na, Rb, Se, and Zn were not affected by external environmental factors or by racial difference; (3) renal and hepatic Cd levels were very high in several cases and the accumulation has still been in progress in the Japanese, whereas the contaminant elements are low in each organ except for lung.     

A comparative study of reactive hyperemia in human forearm skin and muscle

Reactive hyperemia (RH) in forearm muscle or skin microcirculation has been considered as a surrogate endpoint in clinical studies of cardiovascular disease. We evaluated two potential confounders that might limit such use of RH, namely laterality of measurement and intake of non-steroidal anti-inflammatory drugs (NSAIDS). Twenty-three young non-smoking healthy adults were enrolled. In Experiment 1 (n=16), the RH elicited by 3 min of ischemia was recorded in the muscle (strain gauge plethysmography, hand excluded) and skin (laser Doppler imaging) of both forearms. In Experiment 2 (n=7), RH was determined in the dominant forearm only, one hour following oral acetylsalicylic acid (1 g) or placebo. In Experiment 1, peak RH was identical in both forearms, and so were the corresponding durations of responses. RH lasted significantly less in muscle than in skin (p=0.003), a hitherto unrecognized fact. In the skin, acetylsalicylate reduced duration (43 vs. 57.4 s for placebo, p=0.03), without affecting the peak response. In muscle, duration tended to decrease with acetylsalicylate (21.4 vs. 26.0 s with placebo, p=0.06) and the peak increase in blood flow was blunted (27.2 vs. 32.4 ml/min/100 ml tissue with placebo, p=0.003). We conclude that, when using RH as a surrogate endpoint in studies of cardiovascular disease, a confounding by laterality of measurement need not be feared, but NSAIDS may have an influence, although perhaps not on the peak response in the skin.     

Determining heat and mechanical pain threshold in inflamed skin of human subjects

In a previous article in the Journal of Visualized Experiments we have demonstrated skin microdialysis techniques for the collection of tissue-specific nociceptive and inflammatory biochemicals in humans. In this article we will show pain-testing paradigms that are often used in tandem with microdialysis procedures. Combining pain tests with microdialysis provides the critical link between behavioral and biochemical data that allows identifying key biochemicals responsible for generating and propagating pain.Two models of evoking pain in inflamed skin of human study participants are shown. The first model evokes pain with aid of heat stimuli. Heat evoked pain as described here is predominantly mediated by small, non-myelinated peripheral nociceptive nerve fibers (C-fibers). The second model evokes pain via punctuated pressure stimuli. Punctuated pressure evoked pain is predominantly mediated by small, myelinated peripheral nociceptive nerve fibers (A-delta fibers). The two models are mechanistically distinct and independently examine nociceptive processing by the two major peripheral nerve fiber populations involved in pain signaling.Heat pain is evoked with aid of the TSA II, a commercially available thermo-sensory analyzer (Medoc Advanced Medical Systems, Durham, NC). Stimulus configuration and delivery is handled with aid of specific software. Thermodes vary in size and shape but in principle consist of a metal plate that can be heated or cooled at various rates and for different periods of time. Algorithms assessing heat-evoked pain are manifold. In the experiments shown here, study participants are asked to indicate at what point they start experiencing pain while the thermode in contact with skin is heated at a predetermined rate starting at a temperature that does not evoke pain. The thermode temperature at which a subject starts experiencing pain constitutes the heat pain threshold.Mechanical pain is evoked with punctuated probes. Such probes are commercially available from several manufacturers (von Frey hairs). However, the accuracy of von Frey hairs has been criticized and many investigators use custom made punctuated pressure probes. In the experiments shown here eight custom-made punctuated probes of different weights are applied in consecutive order, a procedure called up-down algorithm, to identify perceptional deflection points, i.e., a change from feeling no pain to feeling pain or vice versa. The average weight causing a perceptional deflection constitutes the mechanical pain threshold.     

Marxism and human sociobiology: A comparative study from the perspective of modern socialist economic reforms

Modern socialist economic reforms which center on the establishment of a commodity based economic system, demand a reconsideration of human nature. Marxism and human sociobiology give different answers to questions about human nature, but neither is complete in itself. It seems timely, therefore, to suggest that a combination of biological understanding with a Marxist-based social understanding would produce a more adequate notion of human nature, thereby helping us to resolve a number of problems posed by reforms currently taking place in socialist countries. We might also hope to face new challenges posed in the future.