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Protein structural changes in keratin fibers induced by chemical modification using 2-iminothiolane hydrochloride: a Raman spectroscopic investigation.

For the purpose of investigating in detail the influence of chemical modification using 2-iminothiolane hydrochloride (2-IT) on keratin fibers, the structure of cross-sections at various depths of white human hair, treated with 2-IT and then oxidized, was directly analyzed without isolating the cuticle and cortex, using Raman spectroscopy. In particular, the beta-sheet and/or random coil content (beta/R) and the alpha-helix (alpha) content in human hair fibers were estimated by amide I band analysis. The S-S band intensity, amide III (unordered) band intensity, and beta/R content existing from the cuticle region to the center of cortex region of virgin white human hair remarkably increased by performing the chemical modification using 2-IT. On the other hand, not only the S-S band intensity, but also S-O band intensity existing throughout the cortex region of the bleached (damaged) white human hair increased by performing chemical modification using 2-IT. In particular, beta/R content existing throughout the cortex region of the bleached white human hair decreased, while the skeletal C-C stretch (alpha) band intensity at 935 cm(-1) and the alpha content remarkably increased. This indicates a secondary structural change from the random coil form to the alpha-helix form in the proteins existing throughout the cortex region. From these experiments, we concluded that the formation of new disulfide (-SS-) groups resulting from chemical modification using 2-IT induced the secondary structural changes of proteins existing throughout the cortex region.

Cross-Linking Reagents↗

Reduction mechanism of L-cysteine on keratin fibers using microspectrophotometry and Raman spectroscopy.

In order to investigate the reduction mechanism of L-cysteine (Cys) on keratin fibers, cross-sectional samples of virgin white human hair treated with Cys were prepared. The heterogeneous reaction between Cys and keratin fibers involving the diffusion of Cys into human hair was analyzed at the molecular level using microspectrophotometry and Raman spectroscopy. The diffusion pattern of Cys into human hair showed non-Fickian type characteristics, thus indicating the free amino groups of electrostatically interacted with the anionic ions of the fiber surface. The disconnected relative concentration of -SS- groups at various depths of the hair samples with pH 9.0 was less than the Cys relative concentration, indicating that the reaction rate (the disconnection of -SS- groups) was slower than the diffusion rate of Cys into human hair. From these experiments, we concluded that the free amino groups of Cys electrostatically interacted with the anionic ions of the fiber surface, thereby decreasing the reaction rate (the disconnection of -SS- groups) of Cys at pH 9.0.

China↗

Genetic control of retinal ganglion cell projections.

We have assessed the effects of 15 pigmentation mutations on the development of retinal ganglion cell projections in mice in two ways: (1) by analyzing the pattern of innervation of the ipsilateral lateral geniculate nucleus as mapped in autoradiograms of brains of animals killed 12 days after intravitreal injection of 3H-proline into one eye and (2) by determining the ratio of axonally transported radioactive protein in the contralateral and ipsilateral optic tracts after similar intravitreal injections. Analysis of the ratio of transported protein in the two optic tracts provides a new and useful assay of the degree of decussation in experimental animals. The effects of the mutations on eye pigmentation, whole eye melanin content and relative tyrosinase activity also were examined. The degree of ipsilateral innervation generally correlates with the degree of pigmentation of the retinal pigment epithelium and with tyrosinase activity. However, discrepancies have been found in ch and ce mutants. In these animals the pigment epithelium is well pigmented, and the area of ipsilateral innervation in the lateral geniculate nucleus is extensive, despite a high ratio of label in contralateral to ipsilateral optic tracts and low tyrosinase activity. Furthermore, mice heterozygous for the c2J allele have pigmentation and optic projections that are normal even though tyrosinase is reduced to 40% of normal. The few anomalous results suggest that alternative or additional factors may control optic axon projections.

Alleles↗

New semidominant mutations that affect mouse development.

Dominantly acting mutations that produce visible phenotypes are frequently recovered, either during routine maintenance of colonies or from mutagenesis experiments. We have studied 12 dominant mouse mutations that cause a tail dysmorphology, a coat spotting phenotype, or a combination of these. The majority of these mutations act in a semidominant manner with the homozygous state associated with embryonic lethality and a visible phenotype at or before midgestation. The homozygous phenotypes include axis truncation and neural crest cell defects, as may be expected from the heterozygous phenotypes. The majority of mutations, however, also produced other phenotypes that include neural tube closure defects and aberrant heart looping. In one coat spotting mutant the homozygous condition is lethal before neural crest cell production commences. The mutated genes often function in processes additional to those alluded to by the heterozygous phenotype.

Alkylating Agents↗

Four novel variants in MC1R in red-haired South African individuals of European descent: S83P, Y152X, A171D, P256S.

Skin, hair and eye pigmentation is a polygenic multifactorial trait determined by the cumulative effects of multiple genetic variants and environmental factors. MC1R is one of the genes involved in pigmentation, and has been implicated in the red hair and pale skin trait in human Caucasoid individuals. This study was undertaken to investigate variants in the MC1R gene in Caucasoid individuals in South Africa, who are of European decent. Seven unrelated individuals were studied, all of whom were found to be either homozygous for a single mutation or compound heterozygous for two different mutations. We report four novel MC1R missense mutations: S83P, Y152X, A171D and P256S. This study supports the view that two mutations are necessary, but not necessarily sufficient, to give rise to the red hair and pale skin phenotype.

Codon↗

Common acquired naevi and the risk of malignant melanoma.

The relationship between common acquired naevi and malignant melanoma of the skin was assessed in a case-control study. IN 183 patients and 183 controls matched for sex, age and area of residence, there was a strong association between the presence of pigmented naevi on the arms and melanoma. In comparison with persons having no naevi on the arms, the crude relative risk of melanoma was 28.0 when naevi were present. After adjusting for hair colour, propensity to sunburn and lifetime sun exposure, the final risk estimate was 30.1. Family history did not appear to be a determinant of disease independent of the above risk factors.

Arm↗

The Danish case-control study of cutaneous malignant melanoma. I. Importance of host factors.

The relationship between cutaneous malignant melanoma and possible host factors was investigated in a population-based case-control study from East Denmark over a 3-year period. A total of 474 melanoma patients and 926 population controls aged 20-79 years were interviewed. Patients with lentigo maligna melanoma were not included. The major constitutional risk factors were: number of raised naevi on the arms (RR = 5.1 for 5+ vs. none), degree of freckling (RR = 2.9 for many vs. none), and light hair colour (RR = 1.7 for blond/fair vs. dark brown/black), which were independent of one another. An apparent synergy between number of raised naevi on the arms and degree of freckling was found. Thus, persons at high risk of melanoma may be identified by a simple assessment of naevi and degree of freckling. No significant difference was found between superficial spreading melanoma and nodular melanoma with regard to the most important host factors.

Adult↗

Rescue of the albino phenotype by introduction of a functional tyrosinase gene into mice.

The c-locus of the mouse is thought to encode tyrosinase, the key enzyme for melanin synthesis in melanocytes of the skin and the eye. Recently, a mouse cDNA was isolated and shown to confer tyrosine activity on a cell line which expressed no specialized functions for melanin synthesis. To verify that the isolated tyrosinase gene is encoded at the genetically well characterized c-locus, a minigene was assembled from tyrosinase cDNA and tyrosinase genomic DNA and used for generation of transgenic mice. Following microinjection of this construct into fertilized eggs of an albino mouse strain, transgenic mice were obtained which showed pigmentation in skin and eyes. By in situ hybridization, we show expression of the transgene in melanocytes of the hairbulb and in the pigmented cell layers of the eye. We conclude that we have rescued the albino mutation (c/c) by introduction and expression of a functional tyrosinase gene.

Albinism↗

Multiphoton fluorescence lifetime imaging of human hair.

In vivo and in vitro multiphoton imaging was used to perform high resolution optical sectioning of human hair by nonlinear excitation of endogenous as well as exogenous fluorophores. Multiphoton fluorescence lifetime imaging (FLIM) based on time-resolved single photon counting and near-infrared femtosecond laser pulse excitation was employed to analyze the various fluorescent hair components. Time-resolved multiphoton imaging of intratissue pigments has the potential (i) to identify endogenous keratin and melanin, (ii) to obtain information on intrahair dye accumulation, (iii) to study bleaching effects, and (iv) to monitor the intratissue diffusion of pharmaceutical and cosmetical components along hair shafts.

Adult↗

The development of regional pigmentation patterns in black and tan (at) mice.

Mice which express the black and tan (at)allele at the agouti locus have black dorsal and yellow ventral hairs. To determine the site of action of this allele, the kind of hair pigment produced by various dermal-epidermal recombinations of dorsal and ventral black and tan (at/at), ventral nonagouti (a/a) and ventral yellow (Ay/a) embryonic skin has been investigated. The results indicate that it is the dermis which is responsible for the development of regional pigmentation patterns in black and tan mice.

Alleles↗

Interactions between normal and pigment cell populations mutant at the dominant-spotting (W) and steel (Sl) loci in the mouse.

Through the use of dermal-epidermal recombination methods a competition between mouse embryo melanoblasts of the genotype Wv/w C/C, w/w c/c, Sld/sl C/C and sl/sl c/c was established. Control combinations were made between C/C and c/d components. The extent of pigment found in hair of grafts after three weeks growth in mouse testes was used as evidence of an interaction between populations. Normal and albino melanoblasts were found to be similar in viability, whereas melanoblasts of the genotype Wv/w C/C were largely excluded from hair follicles when placed in competition with w/w c/c melanoblasts. No difference in competitive advantage was observed between Sld/sl C/C and sl/sl c/c populations. These results confirm that the W and sl loci act at different sites. In addition they suggest that Wv/w melanoblasts are marginally viable cells that cannot compete with normal melanoblasts when the popuolations interact. The Wv/w melanoblast failure can also explain the spotting pattern and pigment dilution characteristic of dominant-spotting heterozygous mice.

Alleles↗

Pigment synthesis in the Himalayan mouse.

The effect of temperature on pigment synthesis in adult and juvenile Himalayan mice was investigated. Since pigment synthesis only occurs in actively growing hair, adult mice were plucked to induce hair growth. The extent of darkening of the hair was recorded by photography against a reference scale. The presence of pigment granules in hair follicles was investigated histologically. Housing adult and juvenile mice at 15 degrees C results in the synthesis of pigment in growing hair follicles whereas housing at 30 degrees C results in the absence of pigment granules in the growing hair follicles.

Animals↗

Seasonal variations in hair pigmentation of white-tailed deer and their relationship to sexual activity and plasma testosterone.

Intensity of hair pigmentation of dorsal scrotum, nose, cheek and forehead areas of seven mature, male white-tailed deer were determined from close-up colour slides taken once a month during a 2-year period. Blood samples and skin biopsies from forehead areas were taken at the same time as the photographs. Plasma testosterone (T) levels were measured by radioimmunoassay and T in the skin was investigated by immunohistology. Seasonal variations of hair pigmentation are most pronounced in the forehead region followed by the cheek, scrotum, and nose area. Peak blood levels of T (15.4 ng/ml) were detected in November. The highest correlation between T levels and pigmentation of the forehead area (R = 93%; R2 = 0.87), was established when pigmentation values were shifted two months ahead. Immunohistologically detectable T was localized in hair follicles, hair sheets and apocrine glands but not in the sebaceous glands. It is hypothesized that pigmentation of head regions might serve as a visual cue indicating the sexual status of an individual.

Acclimatization↗

Tyrosinase activity and the expression of the agouti gene in the mouse.

Tyrosinase activity at the time of phaeomelanin synthesis in neonatal mice is lower in agouti than in black skin and hair bulb tissue, and this depressed activity is associated with a reduction in the electrophoretically distinct de novo form of the enzyme. Direct chemical measurements of sulphydryl compounds show elevated levels in agouti hair bulb tissue at this stage of development. The addition of exogenous copper to hair bulb extracts raises the activity of tyrosinase in agouti to approximately the black level but has no affect on black itself. These results are discussed in relation to the role of sulphydryl compounds and copper availability in regulating tyrosinase activity and turnover.

Animals↗

Prolactin-dependent seasonal changes in pelage: role of the pineal gland and dopamine.

The Siberian hamster displays seasonal changes in pelage that are dependent upon fluctuations in circulating prolactin levels. Pinealectomy prevented the decrease in serum prolactin and molt to the winter pelage displayed by castrated males housed under a short-day photoperiod. A dopaminergic antagonist, pimozide, enhanced prolactin levels in both pinealectomized and sham-operated animals under both long and short photoperiods. In the short-day animals, this effect of pimozide was associated with a prevention of the development of winter pelage. These results indicate that seasonal prolactin levels and related pelage changes are dependent upon the integrity of the pineal gland. However, basal prolactin levels under different photoperiod conditions appear to be only partly regulated by the actions of the dopaminergic system.

Analysis of Variance↗

Cold exposure and food restriction facilitate physiological responses to short photoperiod in Djungarian hamsters (Phodopus sungorus).

We investigated the influence of ambient temperature (Ta) and food availability on seasonal timing and extent of physiological responses to short photoperiod (SP), in particular daily torpor, in Djungarian hamsters (Phodopus sungorus). Exposure of hamsters to cold temperature (Ta = 5 degrees C), relative to warm Ta (23 degrees C), resulted in: 1) a significant advance (P < 0.05) of the first occurrence of torpor among cold-exposed hamsters (days 52-97 vs. days 83-99 in SP); 2) a higher (P < 0.01) incidence of torpor (48% vs. 20% torpid animals/day); 3) a higher (P < 0.05) degree of molt into the winter pelt; and 4) an accelerated reduction of body weights (P < 0.001). However, within SP/cold-Ta exposed groups, individual hamsters clearly showed different tendencies for torpor (torpor on 0-95% of days observed). Therefore, we evaluated the effects of small changes in Ta on torpor frequency and extension by subjecting the same SP-adapted individuals to varying temperatures. Lowering of Ta from 15 degrees C to 10 degrees C and 5 degrees C caused significant (P < 0.05) increases in the incidence of torpor (20%, 33%, and 40%, respectively) and lower minimal body temperatures during hypothermia (P < 0.05). When the same animals were subjected to 24-48 h lasting periods of food restriction (60% of the ad libitum intake), torpor frequency further increased 1.8- to 2.6-fold at all Tas. These results show that Ta and food availability are effective in modifying both seasonal timing and extent of photoperiodically controlled adaptations. This integration of multiple environmental cues, combined with a pronounced within-species variability of winter adjustments, indicates that Djungarian hamsters are capable of flexible physiological responses towards unpredictable climatic changes in the environment.

Animals↗

Molting in the Djungarian hamster (Phodopus sungorus Pallas): seasonal or continuous process?

After transfer into a short daylight regimen, the brownish summer pelage of the Djungarian hamster (Phodopus sungorus) changes into the whitish winter phenotype. Although changes in serum prolactin levels are identified as the initiating hormonal signal, morphological data about molting in that species are sparse. The aim of this study was to characterize in detail the summer and winter pelage of the Djungarian hamster and to analyze the alterations in the skin and pelage induced by photoperiodic changes. The main difference between summer and winter hair types is the pattern of pigmentation. In contrast to other mammalian species showing seasonal changes, the winter coat of the Djungarian hamster is not characterized by an increase in hair density. Molting patches were observed at all times, even in the winter coat, showing that the light regimen does not control the process of molting itself but the pattern of pigmentation and eventually the loss of hair during the single molting wave.

Animals↗