Analysis of structural changes in bleached keratin fibers (black and white human hair) using Raman spectroscopy

To investigate the influence of bleaching treatments on keratin fibers, the structure of cross-sections at various depths of bleached human hair (black and white human hair) was directly analyzed without isolating the cuticle and cortex, using Raman spectroscopy. The S-S band intensity existing from the cuticle region to the center of cortex region of virgin white human hair decreased, while the S-O band intensity at 1040 cm(-1), assigned to cysteic acid, increased by performing the bleaching treatment. Especially, the S-O band intensity of the cuticle region increased remarkably compared with that of the cortex region. Also, the amide III (unordered) band intensity in the cortex region increased, indicating that some of the proteins existing throughout the cortex region changed to the random coil form. Moreover, it has been found that the S-S band intensity existing from the cuticle region to the center of the cortex region of the virgin black human hair decreased remarkably, while the S-O band intensity increased significantly compared with that of the virgin white human hair by performing the bleaching treatment. From these experiments, we concluded that the melanin granules including metal ions act as a decomposition accelerator for the oxidizing agent, thereby leading to a higher level of disulfide (-SS-) group cleavage in the black human hair compared with that of the white human hair.     

Morphological changes in hair melanosomes by aging

Various changes appear in hair by aging, and graying is the most remarkable one. Changes in melanocytes have been well studied as the cause; however, little is known about the change in melanosomes which have a role of carrying melanin pigments into hair shafts. Using pigmented hairs of Japanese females from their age of 4–75, I isolated melanosomes and observed them. As a result, I found a significant change in the morphology of hair melanosomes with age. They were ellipsoidal on the whole and there was no age dependence in the major axis, while the minor axis significantly increased and its frequency distribution broadened with age. The anticipated volume of the melanosome of the oldest person hairs was about twice larger than that of child hairs. This enlargement of melanosome seems to be a cause of the age‐related color change in pigmented hairs from brown to black.     

Epidermal patterning and induction of different hair types during mouse embryonic development

An intriguing question in developmental biology is how epidermal pattern formation processes are established and what are the molecular mechanisms involved in these events. The establishment of the pattern is concomitant with the formation of ectodermal appendages, which involves complex interactions between the epithelium and the underlying mesenchyme. Among ectodermal appendages, hair follicles are the “mini organs” that produce hair shafts. Several developmental and structural features are common to all hair follicles and to the hair shaft they produce. However, many different hair types are produced in a single organism. Also, different characteristics can be observed depending on the part of the body where the hair follicle is formed. Here, we review the mechanisms involved in the patterning of different hair types during mouse embryonic development as well as the influence of the body axes on hair patterning. Birth Defects Research (Part C) 87:263–272, 2009. © 2009 Wiley‐Liss, Inc.     

Polarized Raman microspectroscopy on intact human hair

Polarization‐resolved Raman microspectroscopy with near‐infrared laser excitation was applied to intact human hair in order to non‐invasively investigate the conformation and orientation of the polypeptide chains. By varying the orientation of the hair shaft relative to the polarization directions of the laser/analyzer, a set of four polarized Raman spectra is obtained; this allows to simultaneously determine both the secondary structure of hair proteins and the orientation of the polypeptide strands relative to the axis of the hair shaft. For the amide I band, results from a quantitative analysis of the polarized Raman spectra are compared with theoretically expected values for fibers with uniaxial symmetry. Based on the polarization behavior of the amide I band and further vibrational bands, a partial ordering of α‐helical polypeptide strands parallel to the hair shaft can be concluded. We suggest that this microspectroscopic approach may be used for human hair diagnostics by detecting structural or orientational alterations of keratins. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Determination of copper and zinc levels in human hair

The Cu and Zn levels of both 607 men (1–85 y old) and 649 women (1–92 y old) were determined by atomic absorption spectrometry. Sex does not influence Cu (14.89±0.89 μg/g and 15.26±0.79 μg/g hair for males and females, respectively) and Zn contents (200.97±9.68 μg/g for men and 209.81±9.49 μg/g hair for women). Age influences Cu and Zn concentrations, but only significantly in females: Cu levels decrease over 60 y of age; whereas Zn levels increase significantly from age groups 2–5 to 20–40 years. Hair color influences Cu concentrations in both males and females. In males, white hair containes less Cu than black hair; in females, white hair's Cu levels are significantly lower than those of dark blond, red, light brown, and brown hair. There are no significant differences in Zn concentrations with respect to different hair colors, in either males or females.     

Root hair-specific expansins modulate root hair elongation in rice

Root hair growth requires intensive cell‐wall modification. This study demonstrates that root hair‐specific expansin As, a sub‐clade of the cell wall‐loosening expansin proteins, are required for root hair elongation in rice (Oryza sativa L.). We identified a gene encoding EXPA17 (OsEXPA17) from a rice mutant with short root hairs. Promoter::reporter transgenic lines exhibited exclusive OsEXPA17 expression in root hair cells. The OsEXPA17 mutant protein (OsexpA17) contained a point mutation, causing a change in the amino acid sequence (Gly104→Arg). This amino acid alteration is predicted to disrupt a highly conserved disulfide bond in the mutant. Suppression of OsEXPA17 by RNA interference further confirmed requirement for the gene in root hair elongation. Complementation of the OsEXPA17 mutant with other root hair EXPAs (OsEXPA30 and Arabidopsis EXPA7) can restore root hair elongation, indicating functional conservation of these root hair EXPAs in monocots and dicots. These results demonstrate that members of the root hair EXPA sub‐clade play a crucial role in root hair cell elongation in Graminaceae.     

Elemental anomalies in hair as indicators of endocrinologic pathologies and deficiencies in calcium and bone metabolism

Analytical results obtained by ICP-MS of hair samples from a group of women from Rio de Janeiro city show that abnormal Ca and P concentrations in this compartment can be an indication of pathologies affecting the metabolism of these elements. The study was conducted initially on 900 women (outpatients, >40 years). From this group, approx. 24% showed anomalously high or low Ca concentrations in hair, in some cases correlated to anomalies of other elements. In 144 cases (16%), very high concentrations of Ca (up to 8,285 mg/kg) were measured, frequently correlated with a high concentration of P (up to 4,720 mg/kg), exceeding by far the reference intervals for this age/sex group. Follow-up studies of a few individuals from this group gave first indications that their abnormal hair compositions were related to endocrinologic pathologies affecting calcium/bone metabolism. Very low hair Ca-concentrations were observed in older women (72 cases, age >60 years) and related to senile osteoporosis. Complementary investigations of patients with recognized endocrinologic pathologies (hyperthyroidism, hyper- and hypoparathyroidism) and osteomalacia gave statistical support for the hypothesis that hair concentrations of Ca, P and various other trace elements are influenced characteristically by these diseases. In patients with hyperparathyroidism and hyperthyroidism, both elements showed significant increase in hair, whereas patients with rickets/osteomalacia had only elevated Ca concentrations, together with suspiciously high toxic levels of Cd and various other elements (Fe, Mn, Mg, Sr, Ba). Patients with hypoparathyroidism had significantly decreased Ca and P concentrations in hair. Statistical evaluation of these data by multivariant analysis (MANOVA) using a contrast matrix and by discriminant analysis showed that elemental hair anomalies can be used to diagnose correctly the above-mentioned pathologies, demonstrating the usefulness of hair analysis as a complementary tool for the detection of disturbances in calcium/bone metabolism.     

Diagnostic value of hair shafts and squamous cells in peritoneal washing cytology

OBJECTIVE: Little attention has been given to hair shafts and squamous cells in peritoneal fluid. To investigate their diagnostic value in peritoneal washing specimens, we reviewed peritoneal washing cytology preparations from 83 cases of ovarian tumors. STUDY DESIGN: We reviewed peritoneal washing specimens and histologic sections of 86 cases of ovarian tumors and tumorous conditions, including 22 teratomas, 16 serous adenocarcinomas, 10 clear cell adenocarcinomas, 9 endometrioid adenocarcinomas, 5 cases of endometriosis, 4 mucinous adenomas, 3 serous cystadenocarcinomas and 17 other tumors. RESULTS: We observed both squamous cells and hair shafts surrounded by inflammatory cells in 5 of the 22 cases of ovarian teratoma. Rupture of an ovarian teratoma was clinically and histologically found in one of the five cases. Hair shafts were not observed in the other tumors or in nonneoplastic conditions. The diameter of hair shafts in peritoneal washing specimens ranged from 10 to 28.8 microns (average, 16.6), and such hair shafts were present within an ovarian teratoma examined histologically. The diameter of hair shafts from six normal adults who were examined as controls ranged from 61.5 to 118.6 microns (average, 89.4). CONCLUSION: Hair shafts and squamous cells surrounded by inflammatory cells in peritoneal washing specimens are a diagnostic clue to ovarian teratoma and can be observed even when rupture of the tumor is not detected clinically or microscopically.     

Analysis of internal structure changes in black human hair keratin fibers with aging using Raman spectroscopy

To investigate the internal structure changes in virgin black human hair keratin fibers due to aging, the structure of cross-sections at various depths of virgin black human hair (sections of new growth hair: 2 mm from the scalp) from a group of eight Japanese females in their twenties and another group of eight Japanese females in their fifties were analyzed using Raman spectroscopy. For the first time, we have succeeded in recording the Raman spectra of virgin black human hair, which had been impossible due to high melanin granule content. The key points of this method are to cross-section hair samples to a thickness of 1.50-microm, to select points at various depths of the cortex with the fewest possible melanin granules, and to optimize laser power, cross slit width as well as total acquisition time. The reproducibility of the Raman bands, namely the alpha-helix (alpha) content, the beta-sheet and/or random coil (beta/R) content, the disulfide (--SS--) content, and random coil content of two adjoining cross-sections of a single hair keratin fiber was clearly good. The --SS-- content of virgin black human hair from the Japanese females in their fifties for the cortex region decreased compared with that of the Japanese females in their twenties. On the other hand, the beta/R and alpha contents of the cortex region did not change.     

A new method for DNA extraction from feces and hair shafts of the South China tiger (Panthera tigris amoyensis)

It is commonly known that tigers (Panthera tigris) groom themselves by licking their coats, which leads to an abundance of hairs in their feces. These hairs are designated specially as “fecal hairs”. In our study, in order to explore fecal hairs potential as a DNA source for genetic analysis, 55 fecal hair samples were collected from 23 captive South China tigers (P. t. amoyensis). According to the amplification of mitochondrial primers loop F and loop R, DNA quality of noninvasive samples were grouped into three grades: grade I—the highest-quality DNA, grade II—high-quality DNA, and grade III—poor-quality DNA. No failed amplifications on microsatellite primers and only 0.27% genotyping errors occurred with grade I fecal hair DNA, as compared with 9.4% failed amplifications on microsatellite primers and 9.5% genotyping errors with grade II fecal hair DNA. It was found that 25.45% of fecal hair DNA was grade I and 65.45 and 10.00% of fecal hair DNA were grades II and III, respectively, as compared with 4.35% grade I fecal DNA and 34.78 and 60.87% grades II and III fecal DNA, respectively. Thus, higher-quality DNA can be extracted from fecal hairs than feces. In addition, DNA could be extracted from hair shafts of tigers and a minimum of 2000 hair shafts were required for visible DNA bands on a 1% agarose gel. These findings demonstrate that fecal hairs may serve as a convenient and reliable genomic DNA source for genotype analysis. Zoo Biol 28:49–58, 2009. © 2008 Wiley-Liss, Inc.     

Generation and suppression of singlet oxygen in hair by photosensitization of melanin

We have studied the spectroscopic properties of hair (white, blond, red, brown, and black) under illumination with visible light, giving special emphasis to the photoinduced generation of singlet oxygen (¹O₂). Irradiation of hair shafts (λ_ex > 400 nm) changed their properties by degrading the melanin. Formation of C3 hydroperoxides in the melanin indol groups was proven by ¹H NMR. After 532-nm excitation, all hair shafts presented the characteristic ¹O₂ emission (λ_em = 1270 nm), whose intensity varied inversely with the melanin content. ¹O₂ lifetime was also shown to vary with hair type, being five times shorter in black hair than in blond hair, indicating the role of melanin as a ¹O₂ suppressor. Lifetime ranged from tenths of a nanosecond to a few microseconds, which is much shorter than the lifetime expected for ¹O₂ in the solvents in which the hair shafts were suspended, indicating that ¹O₂ is generated and suppressed inside the hair structure. Both eumelanin and pheomelanin were shown to produce and to suppress ¹O₂, with similar efficiencies. The higher amount of ¹O₂ generated in blond hair and its longer lifetime is compatible with the stronger damage that light exposure causes in blond hair. We propose a model to explain the formation and suppression of ¹O₂ in hair by photosensitization of melanin with visible light and the deleterious effects that an excess of visible light may cause in hair and skin.     

Determination of triazolam involving its hydroxy metabolites in hair shaft and hair root by reversed-phase liquid chromatography with electrospray ionization mass spectrometry and application to human hair analysis

A sensitive method using reversed-phase liquid chromatography coupled with electrospray ionization mass spectrometry has been developed for simultaneous determination of triazolam and its hydroxy metabolites in hair. After the addition of deuterium-labeled 1-hydroxymethyltriazolam as an internal standard, the analytes in hair shaft and hair root samples were extracted with a basic medium, CH(2)Cl(2):MeOH:28% NH(4)OH (20:80:2) at room temperature overnight. The chromatographic separation of the analytes was achieved using a semimicro HPLC column (3-microm particle size; 100 x 2.0-mm i.d.) by gradient elution with acetonitrile in water containing 1% acetic acid as eluent. The mass spectrometer was operated in selected-ion monitoring mode at quasi-molecular ions [M+H](+) of triazolam and its metabolites. The method has been applied to determine the incorporation of triazolam and its metabolites into the hair shafts and hair roots of Dark Agouti rats administered 3 or 6 mg/kg triazolam intraperitoneally twice a day for 5 days. Triazolam, 1-hydroxymethyltriazolam, and 4-hydroxytriazolam were incorporated into the hair shafts and the hair roots. The concentration of 4-hydroxytriazolam was the highest of all compounds detected. An unknown substance considered to be 1,4-dihydroxytriazolam also appeared in the hair samples. The structural elucidation was performed with online HPLC-MS after acetylation of the substance with acetic anhydride and pyridine. The time course studies of triazolam and the metabolites in both rat hair roots and plasma were carried out after single intraperitoneal administration of triazolam. The concentrations of triazolam and the metabolites in the hair roots reflected those in the plasma. The proposed method using selected-reaction monitoring was applied to the determination of triazolam and the metabolites in human hairs of a triazolam addict. Triazolam, 1-hydroxymethyltriazolam, and 4-hydroxytriazolam were identified in the black hair shafts, whereas only triazolam was detected in the hair roots and the white hair shafts. This is the first report on the detection of triazolam and its metabolites in human hairs.     

Selenium levels in human plasma and hair in northern poland

The aim of this study was to (1) estimate the concentration of selenium in the plasma of 146 residents (65 men and 81 women) and in the hair of 34 persons from the Gdańsk region in northern Poland, aged 19–70 and (2) compare the obtained results with data corresponding to healthy populations living in different European countries. Selenium in plasma was determined by atomic absorption spectrometry using the hydride generation method. The mean selenium concentration in plasma of the investigated persons was 73.3 ± 14.1 μg/L, 76.7 ± 13.2 μg/L in men, and 70.4 ± 14.7 μg/L in women. No age — dependent differences in plasma selenium were found in the investigated population. In 20% of the investigated persons, the selenium level in plasma was lower than 60 μg/L. The mean selenium concentration in hair was 0.30 ± 0.11 μg/g. A positive, statistically significant correlation between selenium concentrations in the plasma and hair of the investigated persons was found. The obtained results indicate that the selenium level in significant part of this population is suboptimal and should be elevated by supplementation with this element.     

Exosomes derived from uMSCs promote hair regrowth in alopecia areata through accelerating human hair follicular keratinocyte proliferation and migration

Alopecia areata (AA) is a complex genetic disease that results in hair loss due to an autoimmune-mediated attack on the hair follicle. Mesenchymal stem cells (MSCs) have great potential to induce hair regeneration due to their strong secretion ability and multidirectional differentiation. Recent studies have revealed that the therapeutic potential of MSCs comes from their secretion ability, which can produce large amounts of bioactive substances and regulate the key physiological functions of subjects. The secretion products of MSCs, such as vesicles, exosomes, and conditioned media, have significant advantages in preparing of biological products derived from stem cells. Human umbilical cord mesenchymal stem cells (uMSCs) are the best choice for exosome production. uMSCs are obtained from the human umbilical cord. The umbilical cord is easy to obtain, and the efficiency of uMSCs isolation and culture higher than that of obtaining MSCs from bone marrow or adipose tissue. In this study, we investigated the effects of exosomes released from uMSCs in AA mice. In summary, due to easy isolation and cultivation, simple preparation, and convenient storage, it is possible to obtain uMSCs, or uMSCs exosomes for research and clinical treatment.     

Telogen elongation in the hair cycle of ob/ob mice

Alopecia impairs the physical and mental health of patients. We have previously shown that 8-week-old ob/ob mice have no reactivity to depilation, which is a stimulus that induces anagen transition in normal mice, while no hair cycle abnormalities have been reported in other studies until mice reach 7 weeks of age. Therefore, we hypothesized that ob/ob mice have abnormalities in hair cycle progression beyond 7 weeks of age. We examined 6- to 24-week-old ob/ob and 6- to 10-week-old normal mice. After acclimation, the dorsal skin was harvested and the hair cycle phase was identified histologically and immunohistochemically. Normal mice showed catagen–telogen and telogen–anagen transitions at 6 and 8–9 weeks old, respectively. In contrast, the anagen–catagen transition was observed in 7-week-old mice and the telogen phase was maintained from 10 to 24 weeks in most ob/ob mice. These results suggests that ob/ob mice are a possible model animal for telogen effluvium.     

DNA repair investigations in nine Italian patients affected by trichothiodystrophy.

Trichothiodystrophy (TTD) is a rare autosomal recessive disorder characterized by brittle hair, mental and growth retardation, peculiar face, ichthyosis, and in 20% of the reported cases photosensitivity. Cellular photosensitivity due to the same genetic defect present in xeroderma pigmentosum group D (XP-D) has been described in several patients. Nine patients with clinical symptoms diagnostic for TTD have been identified in Italy to date. We report the results of DNA repair investigations performed in cultured fibroblasts from these patients and 8 TTD parents. Survival, DNA repair synthesis and RNA synthesis following UV irradiation were all normal in the 8 TTD heterozygous cell strains. Among the 9 TTD-affected individuals, normal cellular UV sensitivity was observed in the 2 patients without signs of clinical photosensitivity. In contrast, the other 7 TTD cell strains showed a notable reduction in UV-induced DNA repair synthesis (UDS) levels, ranging between 40% and 5-15% of normal values. Complementation analysis indicated that in the repair-deficient TTD cell strains the genetic defect is the same as that present in XP-D cells. The biochemical heterogeneity of the XP-D defect in TTD patients characterized by different degrees of defective UDS results in different patterns of response to the killing effect of UV light in non-proliferating cells.     

Effect of equilibrium pH on the structure and properties of bleach-damaged human hair fibers

Hair proteins are significantly affected by environmental pH. This impact tends to increase with prior hair damage. To understand how pH affects bleached hair properties, we utilized a number of techniques allowing for the determination of hair thermal properties, swelling and water sorption, and dry and wet tensile properties. At pH 5, hair proteins had the best structural integrity, as determined by differential scanning calorimetry and the highest tensile modulus. At pH 10, protein cross-linking density decreased, water content and hair cross-sectional diameter increased. Alkaline treatment, when compared with pH 5, did not reduce intermediate filament conditions (evaluated via enthalpy measurement) nor mechanical property performance in the wet state. In contrast to alkaline-treated hair, bleached hair equilibrated at pH 3 behaved very differently: it contained two different crosslink density zones, was the least stiff in dry and stiffest in wet conditions. Additionally, it absorbed less water and had the lowest diameter because of reduced water binding by protonated carboxyl groups. The pH 3 to 10 did not affect the mechanical strength of bleached hair in dry or wet conditions.     

Improving stable isotopic interpretations made from human hair through reduction of growth cycle error

A recent trend in stable isotopic analysis involves the reconstruction of short-term variations in diet using hair segments. However, bulk hair samples typically contain a growth cycle error, which may conceal or confound the most recently incorporated isotopic information. It is assumed that, at any given time, ～85-90% of scalp hairs are actively growing, while the remaining 10-15% have transitioned into a resting or inactive phase, which lasts up to 4 months before hairs are shed. This study uses growth phase to determine the effects of age, sex, and health status on carbon and nitrogen isotopic ratios of hair analyzed in sequential segments. For this study, we selected archaeological hair samples from 10 individuals from Dakhleh Oasis, Egypt. Isotopic analyses of actively growing hair segments were compared to those for mixed growth phase segments from each individual. These data demonstrate the presence of growth cycle error and show that an understanding of structural-functional relationships is essential for interpreting normal versus pathological changes in hair follicle and fiber production. In situations where diet change and mobility produce variations in an individual's isotopic composition, elimination of positional-temporal error in sequential segment hair analyses can facilitate greater understanding of intraindividual metabolic reactions and changes in hair growth cycles. Phase identification may aid in determining the presence of pathological conditions in individuals, especially in those lacking skeletal indications, and provide a more precise estimation of seasonal dietary patterns, access to changing food resources, and metabolic equilibration to a new locality.     

A study of body hair density and distribution in normal women

The distribution and density of terminal hair growth have been studied in 19 different body regions in 100 normal women of fertile age and 25 normal postmenopausal women. In accordance with earlier investigations an age-dependent variation in hair growth pattern was observed. None of the women in either group had terminal hair growth on the cheek, and on the breast region hair was limited to a small circumareolar area. Whereas facial terminal hair growth appeared to be less common than that reported in earlier studies of women from other countries, the frequency of terminal hair in other regions was found to be significantly higher in the Norwegian women. No difference in hair growth pattern between blondes and brunettes could be detected.     

Sex hormone levels and body hair growth in !Kung San and Kavango men from Namibia

The relation between hair growth and levels of sex hormones in serum and saliva was investigated in 256 !Kung San and Kavango men (ages 18 to 39 years) from Namibia/Southern Africa. Serum concentrations of total testosterone (Tser), 5α-dihydrotestosterone (DHT), and estradiol (E2) as well as the level of bioavailable non-SHBG-bound testosterone in the saliva (Tsal) were determined by radioimmunoassay. The distribution and density of scalp and facial hair as well as the development of terminal hair on the chest, abdomen, pubic area, arms, fingers, and legs were categorized using objective criteria. Covariance analyses revealed marked differences in the distribution of body hair in the San and the Negro sample. This is partly explained by a significant influence of androgen and estrogen levels on the growth of terminal hair. DHT and the ratio DHT/Tser are significantly positively related to midphalangeal hair growth and negatively to pubic hair development. Tsal, the bioavailable fraction of total testosterone, exerts a weak positive influence on the degree of arm and leg hair growth; the most significant positive effect on the growth of abdominal, arm, and leg hair in our samples is caused by E2. The ratio Tser/E2 correlates significantly negatively with the arm and leg hair development and the ratio DHT/E2 with the degree of abdominal, pubic, arm, and leg hair, whereas lower DHT concentrations occur in men with stronger hair development. © 1993 Wiley-Liss, Inc.