Human Hair Roots Research Articles

FACILE IN VITRO GLASS SLIDE LIGHT MICROSCOPY METHOD USING TETRACYCLINE TO VISUALIZE A REPETITIVE PATTERN IN AERIAL PLANT ROOT TIPS FILAMENTOUS NETWORK

The main purpose of this manuscript is to introduce a facile light microscopy methodology to visualize plant roots filaments. In a previous manuscript in vitro experiments on freshly plucked human hair roots documented the commonly used antibiotic Tetracycline (TE) deleterious effect on soft tissue, severe enough to allow for visualization of an underlying filamentous skeleton. In this manuscript, TE was also evaluated in a similar fashion of in vitro experiments, this time aerial plant roots were immersed in liquid Tetracycline. Images and video recordings are presented where plant aerial root tissue cells appeared to interact with Tetracycline, thus allowing for exposure of an underlying filamentous network. These filaments were documented undergoing biosorption of Tetracycline, thus indicating a probable cellulose base. It is emphasized that a literature search showed similar, albeit visually different displays of roots filaments obtained by using a Scanning Electron Microscopy. The method herein introduced could be an adjunct to existing established methodology in root function research. Two salient advantages are identified, firstly that the essential minimal material and equipment is limited to a light microscope, glass slides, chosen biological material, water and powder Tetracycline. Secondly, the speed in obtaining results would offer researchers a preliminary or perhaps a final correct conclusion.

From Hair to iPSCs-A Guide on How to Reprogram Keratinocytes and Why.

Keratinocytes, as a primary somatic cell source, offer exceptional advantages compared to fibroblasts, which are commonly used for reprogramming. Keratinocytes can beat fibroblasts in reprogramming efficiency and reprogramming time and, in addition, can be easily and non-invasively harvested from human hair roots. However, there is still much to know about acquiring keratinocytes and maintaining them in cell culture. In this article, we want to offer readers the profound knowledge that we have gained since our initial use of keratinocytes for reprogramming more than 10 years ago. Here, all hints and tricks, from plucking the hair roots to growing and maintaining keratinocytes, are described in detail. Additionally, an overview of the currently used reprogramming methods, viral and non-viral, is included, with a special focus on their applicability to keratinocytes. This overview is intended to provide a brief but comprehensive insight into the field of keratinocytes and their use for reprogramming into induced pluripotent stem cells (iPSCs). © 2020 The Authors.

TRANSFER OF HUMAN HAIR EXOCUTICLES ONTO GLASS BY LIQUID CATALASE AND WHOLE BLOOD (REPORTING BLOOD CLUMPING FORMATION WHEN CONTACT WITH KERATIN AS POSSIBLE FACTOR IN ATHEROSCLEROSIS)

The genesis of cardiovascular diseases, namely intra-arterial plaques formation leading to heart attacks, strokes and other vascular complication remains unanswered. To date there is accumulated evidence attributing antioxidant activity (toxic materials) of biochemical events in the human body as probable cause for the illness. Experts in the area of cardiovascular diseases have identified a lack of standardization in disease management; therefore, clinical trials comparing antioxidant agents (such as catalase) have been difficult to compare. Experts are also calling for new knowledge in relation to blood antioxidants physiology. Keratin is a chemical present in cellular structures in the body, also present in the human hair root and shaft. This manuscript presents a new finding, which is documentation of a binding capacity of catalase with keratin. Red blood cell are the main carriers of the catalase present in the circulatory system; in this manuscript experiments are introduced in support of a hypothesis adding one additional factor in the genesis of atherosclerosis; which is the protein enzyme catalase ever present in erythrocytes reacting with the keratin present in human arteries via an unknown mechanism.

Mode and Structure of the Bacterial Community on Human Scalp Hair.

Bacterial communities on various parts of the human body are distinct. We were the first to report the existence of a stable bacterial community on human scalp hair and demonstrated that an analysis of its structure by terminal restriction fragment length polymorphism (T-RFLP) is helpful for individual discrimination. However, the ecology of the bacterial community on human scalp hair has not yet been elucidated in detail. We herein investigated the mode, quantity, and phylogeny of bacterial communities on the human hair shaft and root and showed the results obtained from one representative individual. Direct SEM observations of hair, without a pretreatment, confirmed the ubiquitous presence of bacteria-like coccoids and rods on the shaft and root of hair from the human scalp, with 105–106 cells cm−2 of hair and 107 cells cm−2 of hair, respectively. These values corresponded to the 16S rRNA gene copy numbers obtained by qPCR. These numbers were not significantly affected by detergent washing. These results represented those obtained from many individuals with different hair lengths, ages, and gender. The major OTUs on the human scalp hair shaft and root were the same and included two species of Pseudomonas (phylum Proteobacteria), Cutibacterium and Lawsonella (phylum Actinobacteria), and Staphylococcus (phylum Firmicutes). These results suggest that major bacteria on the human hair shaft are indigenous and derived from the hair root.

Bone-forming cells with pronounced spread into the third dimension in polymer scaffolds fabricated by two-photon polymerization.

The main aim of this work was to stimulate bone‐forming cells to produce three‐dimensional networks of mineralized proteins such as those occurring in bones. This was achieved by a novel approach using a specific type of mesenchymal progenitor cells (i.e., primary fibroblast cells from human hair roots) seeded on to polymer scaffolds. We wrote polymer microstructures with one or more levels of quadratic pores on to a flexible substrate by means of two‐photon polymerization using a Ti‐sapphire femtosecond laser focused into a liquid acrylate‐based resin containing a photoinitiator. Progenitor cells, differentiated into an osteogenic lineage by the use of medium supplemented with biochemical stimuli, can be seeded on to the hydrophilic three‐dimensional scaffolds. Due to confinement to the microstructures and/or mechanical interaction with the scaffold, the cells are stimulated to produce high amounts of calcium‐binding proteins, such as collagen type I, and show an increased activation of the actin cytoskeleton. The best results were obtained for quadratic pore sizes of 35 µm: the pore volumes become almost filled with both cells in close contact with the walls of the structure and with extracellular matrix material produced by the cells. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 891–899, 2017.

Duplex PCR-RFLP for the Simultaneous Genotyping of Single Nucleotide Polymorphisms in ADH1B and ALDH2 Genes.

Single nucleotide polymorphisms (SNPs) in ADH1B and ALDH2, which encode alcohol dehydrogenase 1B and aldehyde dehydrogenase 2, respectively, are responsible for ethanol-metabolizing activity and alcohol-related disease. We developed a simple and rapid polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method for the simultaneous genotyping of the genes in a single-tube reaction. Using an alkaline lysate of a human hair root as a template, SNP-containing regions of each gene were PCR-amplified simultaneously. The PCR products were directly applied to double restriction digestion, followed by agarose gel band analysis. The duplex PCR-RFLP allows for the simultaneous and accurate determination of the SNP genotypes of ADH1B and ALDH2 within 1.2 h.

Rapid Naked-Eye Discrimination of Cytochrome P450 Genetic Polymorphism through Non-Crosslinking Aggregation of DNA-Functionalized Gold Nanoparticles.

Involvement of single‐nucleotide polymorphism (SNP) genotyping in healthcare should allow for more effective use of pharmacogenomics. However, user‐friendly assays without the requirement of a special instrument still remain unavailable. This study describes naked‐eye SNP discrimination in exon 5 of the human cytochrome P450 2C19 monooxygenase gene, CYP2C19*1 (the wild‐type allele) and CYP2C19*2 (the variant allele with G681A point mutation). The present assay is composed of allele‐specific single‐base primer extension and salt‐induced aggregation of DNA‐modified gold nanoparticles (DNA–AuNPs). Genetic samples extracted from human hair roots are subjected to this assay. The results are verified by direct sequencing. This study should promise the prospective use of DNA–AuNPs in gene diagnosis.

Simultaneous genotyping of single-nucleotide polymorphisms in alcoholism-related genes using duplex and triplex allele-specific PCR with two-step thermal cycles.

We developed a time- and cost-effective multiplex allele-specific polymerase chain reaction (AS-PCR) method based on the two-step PCR thermal cycles for genotyping single-nucleotide polymorphisms in three alcoholism-related genes: alcohol dehydrogenase 1B, aldehyde dehydrogenase 2 and μ-opioid receptor. Applying MightyAmp(®) DNA polymerase with optimized AS-primers and PCR conditions enabled us to achieve effective and selective amplification of the target alleles from alkaline lysates of a human hair root, and simultaneously to determine the genotypes within less than 1.5 h using minimal lab equipment.

Estrone Sulfate Source of Estrone and Estradiol Formation in Isolated Human Hair Roots: Identification of a Pathway Linked to Hair Growth Phase and Subject to Site-, Gender-, and Age-Related Modulations

The present study investigated the metabolism of estrone sulfate into bioactive estrogens in the human hair root, including the effects of hair growth phase, anatomical site, gender, and age. Healthy male (n = 18) and female (n = 20) subjects were investigated. Growing (anagen) and resting (telogen) hair roots were collected from selected scalp and body sites. Estrone sulfate metabolism in the hair root yielded substantial levels of estrone and estradiol. Estrogen synthesis exceeded that associated with aromatization of androgens in a previous study. In subjects <50 years old, estrogen synthesis in scalp hair was lower in men than in women. Comparable levels of estrogen formation were observed in 1) male and female axillary and pubic hair and 2) male beard hair. These levels were higher than the estrogen levels detected in the in scalp hair of men <50 years old. With increasing age, estrogen synthesis increased in men and decreased in women. In telogen hair from all body sites, the capacity to form estrone from estrone sulfate remained unaffected, whereas the ability to form estradiol decreased by 62% and 86% in men and women, respectively. Estrogen formation from estrone sulfate in sexually dimorphic hair is linked to the hair growth phase and is subject to gender- and age-related modulations. The magnitude of the in situ estrogen synthesis from estrone sulfate and the selective arrest of estradiol synthesis at the end of the hair cycle suggest that this pathway plays a crucial role in the regulation of human hair growth.

DNA damage in hair root cells as a biomarker for gamma ray exposure

The purpose of the present research is to examine whether human hair root cells can be used for dose assessment after in vitro exposure to ionizing radiation. Hair root samples plucked from random head regions were collected from 5 healthy human subjects. Some of these hair samples were used as control and some were irradiated with 0.5–5Gy of gamma ray using a Cs-137 gamma irradiator at a dose rate of 0.14Gy/s. DNA damage (single-strand breaks) was determined in hair root cells of these samples using the comet assay technique. The comet assay parameters, tail length (TL) and tail moment (TM), showed a significant increase (p<.05) in single-strand DNA breaks in hair roots cells of the exposed samples compared to control. A linear dose–effect relationship was observed when tail moment or tail length was plotted against the log of the radiation dose. This research suggests a possible use of human hair root cell DNA damage as a biomarker especially for low dose radiation.

Genetic analysis of the human hair roots as a tool for spaceflight experiments

The use of hair roots as experimental samples has been a research focus for understanding the effects of spaceflight on astronauts, because it has many advantages, one of which is the fact that hair matrix cells actively divide in a hair follicle and sensitively reflect the physical conditions of the human body. In 2009, a research program focusing on the analysis of astronauts’ hairs was initiated to examine the effects of long-term spaceflight on the gene expression and mineral metabolism in the human body. Since the number of samples per astronaut is limited to 5 strands of hairs at each sampling point, due to the ethical viewpoint of astronauts or limited resources in space, it is important to develop an effective method for the molecular analysis of small amounts of hair roots. In this study, mRNA successfully extracted from 1, 5, and 10 hair follicles was amplified and subjected to the DNA microarray analysis to compare the gene expression within subjects. The results indicated that (1) it was possible to perform the genetic analysis on hair samples stored at -80℃, even without a fixation buffer and (2) the newly modified method of mRNA extraction and analysis was effective in detecting differential gene expression in samples containing only 5 hairs. In conclusion, RNA was efficiently extracted from 5 hair roots, which is the same number of hair roots used in the space experiment; therefore, this method can be applied to genetically analyze astronauts’ hair samples.

Single-strand DNA breaks in human hair root cells exposed to mobile phone radiation

Purpose: To analyze the short-term effects of radiofrequency radiation (RFR) exposure on genomic deoxyribonucleic acid (DNA) of human hair root cells.Subjects and methods: Hair samples were collected from eight healthy human subjects immediately before and after using a 900-MHz GSM (Global System for Mobile Communications) mobile phone for 15 and 30 min. Single-strand DNA breaks of hair root cells from the samples were determined using the ‘comet assay’.Results: The data showed that talking on a mobile phone for 15 or 30 min significantly increased (p < 0.05) single-strand DNA breaks in cells of hair roots close to the phone. Comparing the 15-min and 30-min data using the paired t-test also showed that significantly more damages resulted after 30 min than after 15 min of phone use.Conclusions: A short-term exposure (15 and 30 min) to RFR (900-MHz) from a mobile phone caused a significant increase in DNA single-strand breaks in human hair root cells located around the ear which is used for the phone calls.

Single-Tube Genotyping from a Human Hair Root by Direct PCR

We have developed a simple, labor-saving, inexpensive and rapid SNP genotyping method that directly uses a human hair root as the template. This single-tube genotyping method was used to successfully and reliably genotype the ADH1B and ALDH2 polymorphisms using a hair root (without DNA isolation) and the polymerase chain reaction (PCR) enzyme kit KOD FX. Since the DNA extraction step was eliminated, the possibility of sample contamination was considerably decreased. The single-tube SNP genotyping was performed by coupling the PCR enzyme kit with allele-specific primer (ASP)-PCR. In the post-genome era, a simple and inexpensive method for diagnostic analysis is in high demand, and this PCR method with a hair root as a template will be very useful for genetic diagnoses in biological and medical laboratories.

Coenzyme Q10 has anti‐aging effects on human hair

IFSCC Magazine, 11 (2008) (1) 37–42Ubiquinones are the most widespread and therefore best investigated bioquinones. Due to their hydrophobic isoprenoid side chain, ubiquinones can be solubilized in organic solvents or lipids but are insoluble in water. Using a specific emulsifier system it has been possible to deliver positive effects to biological systems also from aqueous formulations. Ubiquinone‐50, also referred to as coenzyme Q10, is well known in cosmetic science and especially in skin care because of its antioxidant activity. It is found in the membranes of peroxisomes, lysosomes, vesicles, the endoplasmic reticulum and notably in the inner membrane of the mitochondrion, where it is an important part of the electron transport chain. Using coenzyme Q10 in the proposed emulsifier system we could show that the molecule not only has relevance as an anti‐aging bioactive in skin care but also has positive effects on the human hair follicle. The hair follicle is a complex mini organ and synthesis of hair keratin, the major component of hair fibers, is an essential prerequisite for the growth of strong and healthy hair. But like all biological systems the hair follicle, the biologically active part of the hair, also undergoes an aging process associated among other things with a decline in certain hair keratins. Due to this age‐related shift in basic structural proteins of the hair shaft, mature hair often becomes fragile and difficult to manage. Therefore it is a challenge for cosmetic science to provide bioactives to fight age‐related changes and maintain a youthful appearance of hair. Using cultivated hair follicle keratinocytes we identified coenzyme Q10 as a potent bioactive that stimulates the gene expression of different hair keratins, especially those which are reduced during aging processes in hair follicles. These results led us to investigate a shampoo and a tonic formulation enriched with coenzyme Q10 in a placebo‐controlled panel study. In a left/right comparison a group of healthy volunteers older than 40 years of age applied the formulations daily for 4 days. Throughout the test period the gene expression of different hair keratins from plucked hair follicles was determined using quantitative polymerase chain reaction techniques. Subsequent statistical analysis revealed an increase in age‐relevant hair keratins in human hair roots treated with coenzyme Q10, thus pointing out the striking benefits of coenzyme Q10 in hair care formulations. We conclude that coenzyme Q10 is an ideal ingredient for hair care formulations, providing anti‐aging properties through activation of specific keratins aligned with the needs of mature hair.

Prestaining Method as a Useful Tool for the Agarose Gel Electrophoretic Detection of Polymerase Chain Reaction Products with a Fluorescent Dye SYBR® Gold Nucleic Acid Gel Stain

Three staining methods using SYBR Gold Nucleic Acid Gel Stain (SYBR Gold) as a fluorescent dye were evaluated for the agarose gel electrophoretic detection of DNA. The methods involve prestain, in-gel stain, and poststain methods. DNA markers and polymerase chain reaction (PCR) products obtained by minisatellite variant repeat-PCR (MVR-PCR) amplification in a D1S8 locus were used as model DNA and practical samples, respectively. Among the three methods tested under the usual electrophoretic conditions, a prestain method using a 10000-fold diluted SYBR Gold solution showed most excellent features regarding cost and rapidity to use with good stainability and resolution over loaded DNA amounts of about 98 ng to 300 ng. The prestain method was found to be applicable to the analysis of DNA in MVR-PCR products from a human hair root.

Identification of two novel clusters of ultrahigh-sulfur keratin-associated protein genes on human chromosome 11

We analyzed two novel clusters of keratin-associated protein (KAP) genes on human chromosome 11 (11p15.5 and 11q13.5) in which we identified two known human KRTAP5 genes, KerA (= KRN1) and KerB, and nine novel KRTAP5 family genes. RT-PCR analysis of these KAP genes showed preferential expression in human hair root, suggesting these gene products are required for hair formation. Based on the deduced amino acid sequences, all these KAP proteins were classified into an ultrahigh-sulfur (UHS) type KAP with high cysteine content (>30 mol%). These KAPs also showed high glycine and serine contents (average 24.30 and 21.13 mol%, respectively), distinguishing from other UHS/HS KAP families located on human chromosomes 17 and 21. Dot-matrix analysis revealed a significant similarity between these two KAP gene clusters. We postulated a mechanism by which these two KAP gene clusters are generated via genomic duplication of a primordial gene cluster followed by genetic modification during evolution.

Rapid Individual Identification by Minisatellite Variant Repeat (MVR)-PCR at D1S8 Locus Using "Exponential Law"

We describe an efficient and simple minisatellite variant repeat mapping by PCR (MVR-PCR) method based on the assignment of the tandem array of 29 bp repeating units into a-type, t-type and 0-type (a rarely appearing unamplified unit), from the first repeat unit position (code position 0) of 293 bp in D1S8 locus. After microchip electrophoresis of PCR product amplified from the target DNA of a human hair root, each rung of the ladder at the position of 293 + 29 n bp (n: code position) was detected and the type of repeating unit was determined, i.e., aa, a0, tt, t0, at and 00. The peak area of the rungs from PCR product decreased with increase in code position. We found for the first time that the logarithmic plots of the peak area against the code position showed a linear relationship, which implied that peak areas decrease exponentially. The present method was successfully applied to identify 37 individuals using only a hair root as a biological specimen. This "exponential law" is expected to be an effective tool in forensic science.

Postmortem Microscopic Changes Observed at the Human Head Hair Proximal End

Only two types of human hair roots (proximal ends) derived from decomposing scalps are reported in the literature. The most common representation of the putrid root includes a postmortem dark root band in published photomicrographs. In this study, 22 cases were reviewed in which there was reliable time of death documentation from medical investigator reports. A review of these cases finds that the most common putrid hair proximal end change does not contain the postmortem root band. Four primary types of hair proximal end postmortem change were identified. This study finds no correlation of time of death with scalp hair proximal end decomposition. In addition two examples are presented that suggest that hair roots do not decompose after fresh removal from the scalp and exposure to the outside elements.

Commentary on Linch CA, Smith SL, Prahlow JA. Evaluation of the Human Hair Root for DNA Typing Subsequent to Microscopic Comparison. J Forensic Sci. 1998; 43(2):305–14

Abstract Commentary on Linch CA, Smith SL, Prahlow JA. Evaluation of the human hair root for DNA typing subsequent to microscopic comparison. J Forensic Sci. 1998; 43(2):305–14.

Ultrastructure of human scalp hair shafts as revealed by freeze-substitution fixation.

Human scalp hair is important as a diagnostic clue to many diseases, in medical jurisprudential investigations, and also as a subject of cosmetic treatments. While many ultrastructural studies of the human hair root including the hair follicle have been reported, few studies have been done on the human hair shaft. We report here the ultrastructure of human scalp hair shafts prepared by a rapid-freezing technique followed by freeze-substitution fixation that allows the observation of fine cell structures. Healthy scalp hair shafts from Japanese females 12-13 years of age were rapid-frozen and then freeze-substituted in OsO4-acetone. In addition, this technique was applied to the study of some changes of the hair shafts (i.e., hair damaged by thioglycolic acid cold permanent waving and white hair). By this method, the hair shaft was rapid-frozen throughout without appreciable ice damage although the hair shaft was nearly 100 microm in diameter. The rapid-freezing technique resulted in excellent preservation of the ultrastructure of the hair shafts: lamellar structures in the cuticle and fine fibrous ultrastructures in the cortex were observed without chemical treatments. Thioglycolic acid treatment affected the ultrastructure of both the cuticle and the cortex. Except for the absence of melanin granules, no significant differences in the ultrastructure were observed between white hair and black hair. The rapid-freezing technique followed by freeze-substitution fixation appears to be the most reliable approach for the morphological evaluation of fully keratinized cells and tissues.