Hair Root Cells Research Articles

Direct multiplex allele-specific PCR amplification for ABO genotyping from whole blood, hair root and buccal cell

BackgroundABO genotyping is an increasingly being used for accurate prediction of blood transfusion, organ transplantation and forensic application. Traditional methods require DNA extract and purification which is time-consuming labor-intensive and limit the trace samples detection. MethodsPresent study developed the multiplex allele-specific polymerase chain reaction (AS-PCR) method to identify the six major genotypes of ABO blood groups in two tubes reaction. The samples of whole blood, buccal cell and hair root from 30 volunteers were examined. ResultsAll genotyping results were interpreted and agreed with the serologic findings. The modified AS-PCR method was able to determine ABO genotyping using direct blood and non-invasive samples including buccal cell and hair root. ConclusionsThe modified technique has a high specific and sensitive, rapid and cost-efficiency of genotyping method which might be valuable for transfusion medicine in a clinical laboratory and personal identification in forensic investigation.

Abstract OT1-05-03: Efficacy and safety of scalp cooling device for prevention of alopecia in patients with breast cancer

Abstract Background Chemotherapy for breast cancer causes alopecia as a side effect. Some patients refuse chemotherapy because of alopecia, resulting in the omission of a standard therapy. It is believed that a scalp cooling device can prevent alopecia by promoting vasoconstriction of the scalp and reducing exposure of the hair root cells to anticancer agents. There are phenotypic differences of the efficacy of a scalp cooling device for alopecia. In fact, a Dutch scalp cooling registry reported that the success rate of scalp cooling was 51% in European women and 33% in Asian women. Therefore, we aimed to investigate the efficacy of scalp cooling device for chemotherapy-induced alopecia among Asian women with breast cancer. Trial design This is a phase II trialto evaluate the efficacy and safety of scalp cooling device for risk reduction of alopecia in women with stage I/II/III breast cancer treated with adjuvant/neoadjuvant chemotherapy in a single institute. Eligibility criteria Women diagnosed with Stage I to III breast cancer who are scheduled to receive preoperative or postoperative adjuvant chemotherapy containing anthracycline and/or taxanes are enrolled. Patients who have blood malignancies (leukemia, non-Hodgkin lymphoma, other systemic lymphoma), and cold allergy, are excluded. Specific aims The primary endpoint is the proportion of patients with Common Terminology Criteria for Adverse Events (CTCAE) grade 0-1 alopecia after the completion of all cycles of chemotherapy (success rate). Secondary endpoints are safety, quality of life, use of wig or cap, and success rates after the completion of all cycles of chemotherapy distinguished by anthracycline(AC) and taxane. The cooling device is the Paxman scalp cooling system. Scalp cooling was performed from 30 mins before initiation until 90 mins (25 min for taxane) after chemotherapy. Pictures of the scalp were taken at the time of the initiation of each course. Statistical methods Successful treatment was defined as the presence of less than 50% of hair-loss area. The sample size was calculated using the Simon method, with a type I error of 10% (two-sided) and a study power of 80%.The expected success rate is 30%, with a threshold success rate of 10%, and the required number of patients was estimated to be 19. Present and target accrual Patient accrual was started in April 2018 and present accrual is 3. We plan to enroll a total of 20 patients in the trial. Citation Format: Kodera A, Ogura K, Hattori A, Yukawa H, Sakaguchi S, Matsuoka A, Tanaka N, Kamimura M, Jibiki N, Naritaka Y, Hirano A. Efficacy and safety of scalp cooling device for prevention of alopecia in patients with breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr OT1-05-03.

First evidence of maternally inherited mosaicism in TGFBR1 and subtle primary myocardial changes in Loeys-Dietz syndrome: a case report

BackgroundLoeys-Dietz syndrome (LDS) is a rare multisystemic disorder characterized by vascular and skeletal abnormalities, with considerable intra- and interfamilial variability.Case presentationWe report the case of an 8-year-old male with clinical features of two distinct genetic disorders, namely LDS, manifesting in the first months by progressive aortic root dilatation, arterial tortuosity, bifid uvula, and inguinal hernias and oculocutaneous albinism (OCA) manifesting by white hair and skin that does not tan, nystagmus, reduced iris pigment with iris translucency, and reduced retinal pigment). We identified previously reported, homozygous mutations of TYR, c.1A > G (p.Met1Val) and heterozygous, missense mutation of TGFBR1, c.1460G > A (p.Arg487Gln). Family history revealed that his mother underwent multiple surgical repairs for recurrent hemorrhage originating from the buccal artery. Molecular studies confirmed a maternally inherited low grade TGFBR1 mutation somatic mosaicism (18% in peripheral blood leukocytes, 18% in buccal cells and 10% in hair root cells). Maternal cardiac investigations revealed peculiar cardiovascular features: mild tortuosity at the aortic arch, dilatation of the proximal abdominal aorta, multiple deep left ventricular myocardial crypts, and dysplastic mitral valve. TGFBR2 germline mosaicism has been described in three fathers of children carrying TGFBR2 mutations but, to the best of our knowledge, no case of maternally inherited TGFBR1 mutation mosaicism has been reported so far.ConclusionsThis case report suggests that individuals with somatic mosaicism might be at risk for mild and unusual forms of LDS but germline mosaicism can lead to full blown picture of the disease in offspring.

Homozygosity for the c.428delG variant in KIAA0586 in a healthy individual: implications for molecular testing in patients with Joubert syndrome

Joubert syndrome (JBTS) is a rare neurodevelopmental disorder with marked phenotypic variability and genetic heterogeneity. Homozygous or compound heterozygous mutations in the KIAA0586 gene on chromosome 14q23 are known to be associated with JBTS-23. The frameshift variant c.428delG is the most frequent KIAA0586 variant reported in JBTS-23; yet, homozygosity of this variant was observed in two patients with JBTS-23. However, homozygosity of the c.428delG variant was recently reported as well in one healthy individual. To clarify whether the frameshift variant c.428delG in KIAA0586 is pathogenic in the homozygous state. Whole-exome sequencing as well as RNA analysis were performed. We identified biallelic mutations, including the variant c.428delG and a splice site variant c.1413-1G>C, in KIAA0586 in two siblings with clinical and MRI features of JBTS. The c.1413-1G>C variant was inherited from the healthy father. The c.428delG variant was found in the healthy mother in a homozygous state in blood lymphocytes, hair root cells and buccal epithelial cells. RNA analysis revealed that the transcript harbouring the c.428delG variant was expressed in blood cells from the healthy mother, indicating that transcripts harbouring this variant elude the mechanism of nonsense-mediated mRNA decay. Considering this and the high allele frequency of 0.003117 in the gnomAD database, we conclude that c.428delG represents a JBTS disease-causing variant only if present in compound heterozygous state with a more severe KIAA0586 variant, but not in a homozygous situation.

Numerical simulation of the hair formation –modeling of hair cycle

In the recent years, the fields of study of anti-aging, health and beauty, cosmetics, and hair diseases have attracted significant attention. In particular, human hair is considered to be an important aspect with regard to an attractive appearance. To this end, many workers have sought to understand the formation mechanism of the hair root. However, observing growth in the hair root is difficult, and a detailed mechanism of the process has not yet been elucidated. Hair repeats growth, retraction, and pause cycles (hair cycle) in a repetitive process. In the growth phase, hair is formed through processes of cell proliferation and differentiation (keratinization). During the retraction phase, hair growth stops, and during the resting period, hair fall occurs and new hair grows. This hair cycle is believed to affect the elongation rate, thickness, strength, and shape of hair. Therefore, in this study, we introduce a particle model as a new method to elucidate the unknown process of hair formation, and to model the hair formation process accompanying the proliferation and differentiation of the hair root cells in all three dimensions. In addition, to the growth period, the retraction and the resting periods are introduced to realize the hair cycle using this model.

Persistence of immersed blood and hair DNA: A preliminary study based on casework

In some cases, evidence is collected from rivers, canals, lakes or sink pipes. To determine the utility of analyzing these samples and for cases in which DNA was recovered from submerged bulletproof vest parts, we evaluated the time necessary to degrade the blood and, subsequently, DNA on bulletproof vests. In a second experiment, also based on cases, blood was diluted in water from a kitchen sink pipe and incubated at room temperature for different times. Subsequently, DNA quality was assessed. In a parallel experiment, hair roots were incubated in spring water for different time periods.This study demonstrates that after one week of immersion of the bulletproof vest parts in a canal only one sample from more than 100 samples gave a partial genetic profile. No genetic profile were obtained for the 99 other samples. After one month immersion and despite the finding that blood remained detectable on bulletproof vest parts, no genetic profile was obtained for all samples using the classical STR approach. For longer immersion times, no genetic profiles were obtained.In sink pipe water, an incubation time of 72 h (h) was necessary before significant blood degradation occurred. Nevertheless, high inter-sample variability was observed. This high variability may be explained by the variability of water composition coming from nine different sink pipes.For hair root cells incubated in water, we observed that more than 90% of the DNA was degraded after 72 h.

Effects of bright light exposure during daytime on peripheral clock gene expression in humans.

Light is the strongest synchronizer controlling circadian rhythms. The intensity and duration of light change throughout the year, thereby influencing body weight, food preferences, and melatonin secretion in humans and animals. Although the expression of clock genes has been examined using human samples, it currently remains unknown whether bright light during the daytime affects the expression of these genes in humans. Therefore, we herein investigated the effects of bright light exposure during the daytime on clock gene expression in the hair follicular and root cells of the human scalp. Seven healthy men (20.4±2.2years old; 172.3±5.8cm; 64.3±8.5kg; BMI 21.7±3.1kg/m2, mean±SD) participated in this study. Subjects completed 3-day experimental sessions twice in 1month during which they were exposed to bright and dim light conditions. The mRNA expression of Per1-3, Cry1-2, Rev-erb-α (Nr1d1), Rev-erb-β (Nr1d2), and Dec1 was analyzed using branched DNA probes. No significant changes were observed in the expression of Per1, Per2, Per3, Cry1, Cry2, Rev-erb-α (Nr1d1), or Dec1 following exposure to bright light conditions. However, the expression of Rev-erb-β (Nr1d2) tended to be stronger under bright light than dim light conditions. These results suggest that the bright light stimulus did not influence the expression of clock genes in humans. Long-lasting bright light exposure during the daytime may be required to change the expression of clock genes in humans.

Is the root-colonizing endophyte Acremonium strictum an ericoid mycorrhizal fungus?

In previous investigations, we found that Acremonium strictum (strain DSM 100709) developed intracellular structures with similarity to mycelia of ericoid mycorrhizal fungi in the rhizodermal cells of flax plants and in hair roots of Rhododendron plantlets. A. strictum had also been isolated from roots of ericaceous salal plants and was described as an unusual ericoid mycorrhizal fungus (ERMF). As its mycorrhizal traits were doubted, we revised the hypothesis of a mycorrhizal nature of A. strictum. A successful synthesis of mycorrhiza in hair roots of inoculated ericaceous plants was a first step of evidence, followed by fluorescence microscopy with FUN(®)1 cell stain to observe the vitality of the host cells at the early infection stage. In inoculation trials with in vitro-raised mycorrhiza-free Rhododendron plants in axenic liquid culture and in greenhouse substrate culture, A. strictum was never observed in living hair root cells. As compared to the ERMF Oidiodendron maius and Rhizoscyphus ericae that invaded metabolically active host cells and established a symbiotic unit, A. strictum was only found in cells that were dead or in the process of dying and in the apoplast. In conclusion, A. strictum does not behave like a common ERMF-if it is one at all. A comparison of A. strictum isolates from ericaceous and non-ericaceous hosts could reveal further identity details to generalize or specify our findings on the symbiotic nature of A. strictum. At least, the staining method enables to discern between true mycorrhizal and other root endophytes-a tool for further studies.

Age-Related Hearing Loss and Degeneration of Cochlear Hair Cells in Mice Lacking Thyroid Hormone Receptor β1.

A key function of the thyroid hormone receptor β (Thrb) gene is in the development of auditory function. However, the roles of the 2 receptor isoforms, TRβ1 and TRβ2, expressed by the Thrb gene are unclear, and it is unknown whether these isoforms promote the maintenance as well as development of hearing. We investigated the function of TRβ1 in mice with a Thrb(b1) reporter allele that expresses β-galactosidase instead of TRβ1. In the immature cochlea, β-galactosidase was detected in the greater epithelial ridge, sensory hair cells, spiral ligament, and spiral ganglion and in adulthood, at low levels in the hair cells, support cells and root cells of the outer sulcus. Although deletion of all TRβ isoforms causes severe, early-onset deafness, deletion of TRβ1 or TRβ2 individually caused no obvious hearing loss in juvenile mice. However, over subsequent months, TRβ1 deficiency resulted in progressive loss of hearing and loss of hair cells. TRβ1-deficient mice had minimal changes in serum thyroid hormone and thyrotropin levels, indicating that hormonal imbalances were unlikely to cause hearing loss. The results suggest mutually shared roles for TRβ1 and TRβ2 in cochlear development and an unexpected requirement for TRβ1 in the maintenance of hearing in adulthood.

Recent microwave research studies of Gazi Biophysics & Gazi Non-Ionizing Radiation Protection Center

Recent microwave research studies of Gazi Biophysics & Gazi Non-Ionizing Radiation Protection Center

Scalp cooling to prevent alopecia after chemotherapy can be considered safe in patients with breast cancer

With modern scalp cooling equipment cytotoxic damage of hair root cells can be prevented in half of the patients with cancer at high risk of alopecia. However, traditionally doubt has existed whether scalp cooling might facilitate hiding and disseminating scalp skin metastases and thus decrease survival. We discuss this risk using frequency data on metastases in breast cancer from observational and autopsy studies and the Munich cancer registry. They showed the incidence of scalp skin metastases to be very low and not differ between scalp-cooled (0.04–1%) and non scalp-cooled (0.03–3%) patients with breast cancer and in need of chemotherapy. We found it rather unlikely that the incidence of scalp skin metastases might increase at all after scalp cooling, whereas a very small proportion of patients receiving chemotherapy are at risk to develop metastases at this site. Scalp cooling can thus safely be offered to patients treated with alopecia-inducing chemotherapy.

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.

Argyrophilic nucleolar organizing region associated protein synthesis in hair root cells of humans at different developmental stages and sex

Argyrophilic nucleolar organizing region (AgNORs) associated proteins are important for cell proliferation and various diseases. We investigated AgNOR protein synthesis in hair root cells of males and females at different ages using two-dimensional image analysis. Experiments were performed on 58 healthy male and 24 healthy female volunteers in three groups according to age and sex. Hair root cells obtained from hair follicles were stained with silver. Total AgNOR number/total nuclear number (TAN/TNN) and total AgNOR area/nuclear area (TAA/NA) for each nucleus were analyzed. The only significant difference was observed in TAA/NA values for males and females from 6 to 12 years old. We suggest that the difference is due to high NOR activity caused by increased growth hormone production in hair root cells.

Improved method of differentiation, selection and amplification of human melanocytes from the hair follicle cell pool

Hair root harbours a complex cell pool with an immense developmental potential. Several lineages, including skin, can be differentiated from the multipotent to pluripotent cells of outer root sheath (ORS) of hair follicle. Outer root sheath presents the most opulent non-invasively gained adult stem cell source known. For the purposes of cultivating melanocytes designated for graft-based treatments of depigmentation disorders, we have established an ex vivo/in vitro cultivation method by introducing several methodological improvements to the ORS explant method of Dieckmann. As a result, we gained a higher, purer yield of differentiated melanocytes in half the time (at least 10(6) of 95% pure cells in 4 weeks). This reliable cultivation procedure begins with the epilation of 60 hairs and yields high numbers of ORS melanocytes that could be used for grafting applications. The procedure not only utilises the developmental potential of hair root cell pool and favors differentiation into melanocytes, but also contributes to the general trend of minimal-to-non-invasive strategies for regenerative medicine.

Is there any relationship between decreased AgNOR protein synthesis and human hair loss?

Argyrophilic nucleolar organizing region associated proteins (AgNORs) play roles in cell proliferation and a variety of diseases. We attempted to determine whether decreased NOR protein synthesis causes human hair loss. We studied 21 healthy males who suffered hair loss on the frontal/vertex portion of the head. Hair root cells from normal and hair loss sites were stained for AgNOR. One hundred nuclei per site were evaluated and the AgNOR number and NORa/TNa proportions of individual cells were determined using a computer program. The cells from normal sites had significantly higher AgNOR counts than those from hair loss sites. Also, the cells from the normal sites had significantly higher NORa/TNa than cells from the hair loss sites. In the normal sites, the cells demonstrated more NOR protein synthesis than cells in hair loss sites. Therefore, decreased NOR protein synthesis appears to be related to hair loss in humans.

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.

Root Colonization by Ericoid Mycorrhizae and Dark Septate Endophytes in Organic and Conventional Blueberry Fields in Michigan

Ericoid mycorrhizae are interfaces for nutrient exchange between specialized fungi and roots of blueberries and other ericaceous plants. Dark septate endophytes also associate with roots of the Ericaceae and many other plant families, but the nature of the interaction is not well understood. We measured ericoid mycorrhizae and dark septate endophyte colonization in organic and conventional blueberry fields matched by soil series and, if possible, cultivar and field age. The percentage of hair root cells with ericoid mycorrhizae was generally higher in organic than conventional fields, while dark septate endophyte colonization was higher in sand than muck soils. Across both muck and sandy soil types, ericoid mycorrhizal colonization was negatively correlated with field age, and dark septate endophyte colonization was negatively correlated with soil carbon, total soil nitrogen, and soil ammonium levels. When only sandy soils were included in the analysis, ericoid mycorrhizal colonization was positively correlated with soil pH and hair-root diameter, and negatively correlated with field age, soil ammonium levels, and total soil nitrogen. On sandy soils, dark septate endophyte colonization was not correlated with any measured variable. The functions of ericoid mycorrhizae and dark septate endophytes in commercial blueberry fields require further investigation.

Impact of alopecia and scalp cooling on the well‐being of breast cancer patients

Chemotherapy-induced alopecia, a common and distressing side effect of chemotherapy, may be prevented by scalp cooling, which reduces toxicity of cytostatics in hair root cells. This is the first study designed to assess the effect of scalp cooling on well-being. A prospective multi-centre study was performed in 13 hospitals. Breast cancer patients treated with (N=98) and without (N=168) scalp cooling completed questionnaires (EORTC QLQ-C30 and EORTC-QLQ-BR23, BIS, MBA, HADS) before chemotherapy, and three weeks and six months after the last chemotherapy cycle. Scalp cooling was effective in 52% of the cases. Alopecia was considered among the most distressing problems at all three moments of measurement. A trend towards higher well-being was found in successfully scalp-cooled patients, as indicated by a general better health-related quality of life and better body image, whereas unsuccessfully scalp-cooled patients reported lowest well-being. Scalp cooling contributes not only to the well-being of successfully scalp-cooled patients but also seems to cause additional distress when patients lose their hair despite scalp cooling. This might be related to disappointment due to alopecia despite scalp cooling or possibly to a general higher biological availability of cytostatics. We recommend additional support for patients when scalp cooling is not successful and to spend more effort to maximise the effectiveness of scalp cooling.

A case of chimerism in a paternity study

The STR profile of a phenotipically normal woman typed from blood tissue in a paternity case, revealed more than two alleles in some autosomal loci, result that was interpreted as a chimera due to the coexistence of two genetically distinct cell populations. The aim of this work was to solve the paternity study, elucidate the origin of the chimera, and evaluate its consequence in forensic genetics. Buccal epithelium and hair root tissues showed a single autosomal STR profile. Instead, blood tissue exhibited chimerism in seven out of the twenty autosomal loci tested. The paternity analysis was resolved comparing the unique STR pattern typed from buccal and hair root cells. The detection in blood tissue of the Y allele in Amelogenin locus and typing of a Y-chromosome haplotype confirmed the presence of male cells in the chimeric woman. The deduced male profile cannot be excluded as a son of the alleged father. Based on the results we interpret that the woman subject of the study is a partial or a whole-body chimera. Chimerism may be a pitfall in forensic investigations like paternity testing and crime cases.

Paternal somatic mosaicism of a TGFBR2 mutation transmitting to an affected son with Loeys–Dietz syndrome

We report on somatic mosaicism of a TGFBR2 missense mutation, c.1336G>A (D446N). The affected son with the heterozygous mutation was previously reported [Sakai et al. (2006); Am J Med Genet A 140A:1719-1725]. Further evaluation indicates his clinical condition is Loeys-Dietz syndrome. Parental blood samples were studied to confirm whether the propositus' mutation was a de novo change, and suggested a trace of the mutation in the father. DNAs extracted from blood leukocytes, buccal cells, hair root cells, and nails in the father indicated 52%, 25%, 0%, and 35% of cells harbored the mutation, respectively. This is the first detailed report of somatic mosaicism of a TGFBR2 mutation.