Nail Clippings Research Articles

Clustering of hepatitis C infection among family members in Aden, Yemen

BackgroundHepatitis C infection is a common cause of liver diseases such as cirrhosis and liver carcinoma. This study aims to determine the clustering of HCV infection among family members of HCV-positive patients (index cases) and the association between internal and external risk factors and HCV infection among these family members.MethodsThis cross-sectional study was conducted during a period from January to June 2024. One hundred and seventeen family members belonging to 26 index cases were enrolled in this study. Blood samples were collected from all family members, and then sera were separated and tested for anti-HCV antibodies by using a commercially available Cobas technique based on ECLIA.ResultsAmong 117 family members, the majority 55.6% were females. The clustering of HCV infection among family members was 7.7%. The highest rates were 9.6% among males and 11.8% among members who were in close contact with female-positive cases, 12.8% in the age group 20–29 years, and 27.3% among brothers, but there were no HCV-positive cases detected among fathers, daughters, and husbands. There was a significant association between HCV infection and some behaviors of family members such as sharing the same sleeping places, nail clippers, and towels with index cases and exposure to cupping (p = 0.0001, 0.002, 0.017, and 0.050), respectively.ConclusionThe HCV infection among family members in Aden, Yemen, was low in comparison with most studies globally. The highest rates were found among males, those in contact with female index cases, in the age group 20–29 years, and brothers as relatives, but there were no positive cases among fathers, husbands, daughters, and other relatives.

Portable X-ray fluorescence of zinc and selenium with nail clippings-Mother and Infant Nutrition Investigation (MINI).

Zinc and selenium are essential minerals for human nutrition. Reliable biomarkers of zinc status and selenium status in humans are therefore important. This work investigates a novel portable X-ray fluorescence (XRF) method with the ability to rapidly assess zinc and selenium in nail clippings. This approach used a mono-energetic X-ray beam to excite characteristic X-rays from the clippings. Nail clippings were obtained from the Mother and Infant Nutrition Investigation (MINI), a study designed to assess nutrition in a population of women and their breastfed children in New Zealand. Twenty mother-infant pairings were selected to provide nail clippings at two time points (visit 1 at 3 months postpartum; visit 2 at 6 months postpartum). Nail clippings from each mother-infant pairing were divided into three groupings of clippings prior to analysis: those obtained from a big toe of the mother, those from the other toes of the mother, and those from the toes and fingers of the infant. Clippings were prepared and mounted prior to XRF measurement, providing four distinct fragments from each clipping grouping. These fragments were assessed by XRF using a measurement time of either 300 s (visit 1) or 180 s (visit 2). XRF results were determined through both an automated system output and an analysis of the X-ray energy spectrum. Following this assessment of zinc and selenium with the non-destructive XRF method, clippings were measured for zinc and selenium concentration using a "gold standard" technique of inductively coupled plasma mass spectrometry (ICP-MS). Mean ICP-MS concentrations ranged from 122 μg/g to 127 μg/g for zinc, and from 0.646 μg/g to 0.659 μg/g for selenium. Precision, assessed by a relative standard deviation of measurement, was superior for ICP-MS relative to XRF. For both zinc and selenium, XRF results were compared with ICP-MS concentrations. Linear equations of best fit were determined for each comparison between XRF and ICP-MS results. Coefficients of determination (r2) were stronger for zinc (from 0.74 to 0.95) than selenium (from 0.53 to 0.70). A decrease in XRF measurement time from 300 s to 180 s did not appear to adversely affect the correlation between XRF and ICP-MS results. Using the mono-energetic portable XRF method, the correlation of XRF zinc results with ICP-MS zinc concentrations was improved over previous findings, and selenium measurement was reported for the first time. The method may prove useful for future applications to trace element analysis using nail clippings as a biomarker.

Potential of nail cortisol for welfare assessment in shelter and owned cats

Shelter cats often experience considerable stressors, such as confinement and unfamiliar surroundings, leading to welfare challenges. Evaluating and enhancing their well-being is crucial, and measuring cortisol levels in cats’ nails offers a minimally invasive method for assessing stress and welfare. This study compared nail cortisol concentrations (NCCs) between owned cats (OC), and shelter cats in favorable conditions (FS) and unfavorable conditions (UFS) in Thailand and Vietnam. Nails from 31 OC, 29 FS, and 27 UFS cats were collected twice, approximately 28 days apart, with details of living conditions recorded at the second collection. Nail clippings were processed for NCC measurement using the enzyme-linked immunosorbent assay (ELISA) technique. NCC levels of UFS cats were significantly higher than those of FS and OC cats in both collections, with no gender-based differences. NCCs positively correlated with dog exposure, and negatively correlated with space allowance, number of litterboxes, free-roaming time, air conditioning, and the presence and variety of enrichment (hiding places, elevated areas, outdoor views, scratching structures, and toys). These findings indicate that improved living conditions positively impact welfare, as indicated by NCCs. Nail cortisol can serve as a minimally invasive method for assessing cat welfare, particularly considering long-term factors, and offers valuable insights into the well-being of cats in various conditions.

Prevalence, aetiology and factors associated with onychomycosis among patients with diabetes at a Tertiary Care Centre in Galle, Sri Lanka

Introduction: Epidemiological data of onychomycosis, a common fungal nail infection found in diabetics, is scarce in Sri Lanka. Objective: To determine the prevalence, aetiological agents and associated factors of onychomycosis among patients with clinically suspected diabetics in southern Sri Lanka. Method: A cross-sectional study was done using a sample of 97 diabetic patients suspected of having onychomycosis in a tertiary diabetes care centre in Galle. Nail clippings were collected, and microscopic and cultural examinations were done. Both microscopic and culture positive cases were considered as having onychomycosis. Results: The prevalence of onychomycosis was 18.6% (95% CI: 11.38%, 27.73%). Non-dermatophyte moulds (NDMs) (55.7%) and yeast (30.9%) were the dominant aetiological agents. The leading pathogens found among NDMs were Aspergillus sp. (35.1%) and Fusarium sp. (1.4%) and Candida albicans (8.2%). Aspergillus niger (20.6%) was the most frequently isolated Aspergillus sp. in the NDMs. Frequent use of detergents and walking bare foot were the significant risk factors of onychomycosis. Only 15.5% of the participants were aware of onychomycosis. Conclusion: Onychomycosis is prevalent to a considerable level among diabetics. Regular dermatological investigations of nails and health education are needed to combat the disease. Large scale research is warranted to confirm these assertions.

Biospecimens in the HEALthy Brain and Child Development (HBCD) Study: Rationale and protocol

The HEALthy Brain and Child Development (HBCD) Study, a multi-site prospective longitudinal cohort study, will examine human brain, cognitive, behavioral, social, and emotional development beginning prenatally and planned through early childhood. The longitudinal collection of biological samples from over 7000 birthing parents and their children within the HBCD study enables research on pre- and postnatal exposures (e.g., substance use, toxicants, nutrition), and biological processes (e.g., genetics, epigenetic signatures, proteins, metabolites) on neurobehavioral developmental outcomes. The following biosamples are collected from the birthing parent: 1) blood (i.e., whole blood, serum, plasma, buffy coat, and dried blood spots) during pregnancy, 2) nail clippings during pregnancy and one month postpartum, 3) urine during pregnancy, and 4) saliva during pregnancy and at in-person postnatal assessments. The following samples are collected from the child at in-person study assessments: 1) saliva, 2) stool, and 3) urine. Additionally, placenta tissue, cord blood, and cord tissue are collected by a subset of HBCD sites. Here, we describe the rationale for the collection of these biospecimens, their current and potential future uses, the collection protocol, and collection success rates during piloting. This information will assist research teams in the planning of future studies utilizing this collection of biological samples.

Sensitivity, specificity, and discordance with self-report of nail sample testing for alcohol and cannabis

BackgroundNails accumulate the alcohol metabolite, ethyl glucuronide (ETG), and the cannabis metabolite, carboxy- delta-9-THC over 3–6 months. Few studies have examined nail toxicology testing’s sensitivity and specificity and the agreement between nail testing and self-reported alcohol and marijuana use. MethodsIn an ongoing clinical trial, 1101 veterans completed initial telephone questionnaires and were then asked to mail nail clippings for substance use analysis. We examined sensitivity and specificity of ETG and carboxy- delta-9-THC in nails compared to self-report of alcohol use patterns (the AUDIT-C) and substance-related harms (alcohol and THC subscales of the ASSIST). We then examined factors associated with discordance between nails and self-report. ResultsAlmost two-thirds (707/1101) of respondents mailed in nail clippings. Those with returned nails were disproportionately married, white race, older, and less depressed. At a threshold of 8pg/mg, sensitivity was only.50 to detect risky alcohol use and.49 to detect alcohol-related issues. Sensitivity for marijuana issues was only.61. Specificity was greater than.77 for all measures.Factors associated with positive nails/negative self-report (i.e. false positives) for risky alcohol use on the Audit-C included more pain and being unmarried; false positive nails for alcohol-related issues on the ASSIST were associated with being unmarried and non-Hispanic ethnicity. False positive nails for THC-related issues on the ASSIST were associated with being African American, Hispanic, and having had legal issues. ConclusionsAt standard cut-offs, nail measures had low sensitivity and higher specificity. The groups who disproportionately submit positive nails/negative self-report could have substance use patterns not adequately captured by self-report, inaccurate self-report due to social pressures, or distinct drug metabolism.

Onychomycosis: Prevalence, Fungal Pathogens, Risk Factors, and Antifungal Susceptibility Profile among People Living with Diabetes Mellitus in Kano, Northwestern Nigeria

Abstract Context: Onychomycosis is a fungal infection of the nail unit that over time results in dystrophy and disfigurement. It affects the quality of life and increases the prevalence and severity of foot ulcers in patients with diabetes. Aim: The study investigated the prevalence of onychomycosis, the associated fungal pathogens, and their antifungal susceptibility profile in a cohort of people living with diabetes (PLWDM) in Kano, Nigeria. Settings and Designs: This cross-sectional hospital-based study recruited and enrolled 300 PLWDM from two major diabetes treatment centers in Kano. Materials and Methods: Participants were screened for thickened, brittle, crumbly, or ragged nails, and 30 met the inclusion criteria. Sociodemographic data, clinical history, and other risk factors for onychomycosis were also evaluated. Toenail and fingernail, nail clippings, and scrapings were collected and cultured on Sabouraud dextrose agar (SDA). The identification of yeasts and mold isolates was performed using gram stain, germ tube test, CHROMagar, lactophenol cotton blue (LPCB) mount, and slide cultures. Antifungal susceptibility testing was achieved by the agar well-diffusion method. Statistical Analysis Used: The data generated were analyzed using descriptive statistics and associations between variables checked by the chi-square test or Fisher’s exact test. Results: The prevalence of onychomycosis was 63.3% (19/30), and fingernails were mostly affected. Fungal pathogens recovered include dermatophytes, yeasts, and non-dermatophyte molds, with a predominance of Aspergillus spps. (7/19 (36.8%)), Trichophytonmentagrophytes (4/19 (21.1%)), Trichophytonrubrum (4/19 (21.1%)), and Candidaalbicans (2/19 (10.5%)). A preponderance of the isolates shows a good susceptibility profile to itraconazole, ketoconazole, and clotrimazole, but demonstrated high resistance to fluconazole. Age and use of oral hypoglycemic agents were some of the risk factors identified. Conclusion: The study shows a high prevalence of onychomycosis among PLWDM in Kano and calls for routine screening to minimize or eliminate all possible complications.

Expanding the Frontiers of nail product evaluation: Novel application of differential scanning calorimetry (DSC) for assessing crosslinking density and predicting nail brittleness and flexibility.

While modern industry advancements have expanded nail beautification options, scientific literature primarily focuses on nail biology and medicine, with limited attention on cosmetic treatments. This study aimed to investigate human nail denaturation properties, including gender impact, blending nails to enlarge the sample pool, nail sensitization through bleaching, and active effectiveness testing. The objective was to understand the DSC and bending fatigue relationship, and define the consumer relevance of the DSC test. Nail clippings were collected from adult female and male volunteers. The wet DSC was employed to validate sample preparation, explore the effects of gender, and assess the potential of using blended nails for claims substantiation testing. Nails were sensitized through bleaching using hydrogen peroxide. The effects were confirmed through DSC and nail flexure tests. Furthermore, the ability of actives to address concerns related to nail softness and brittleness was assessed using these techniques. The results confirmed the viability of equilibrating nails in water for up to 14 h as a standardized testing method. The denaturation temperature results were independent of gender and suitable for claims substantiation testing. Blending nails from different sources did not yield significant variations in denaturation properties. A preliminary study suggested that cadaver nails should be used with caution because they exhibited differences in denaturation temperature, influenced by the sampling location. Bending fatigue tests highlighted the significance of humidity, with higher humidity conditions (80%) enhancing nail flexibility and providing better resolution for claims substantiation. Sensitizing the nails with hydrogen peroxide induced alterations in both DSC and bending fatigue results. Proof-of-principle studies demonstrated an elevation in denaturation temperature and a decrease in the number of cycles to break, indicating a nail-hardening effect when formaldehyde was applied. The use of a nail softener led to an enhancement in nail fatigue resistance due to a notable reduction in nail crosslinking density. The measurement of crosslinking density proved to be a sensitive tool for assessing the effects of cosmetic treatments on nails, particularly in predicting outcomes related to nail brittleness and flexibility.

Hepatitis B Virus Prevalence and Transmission in the Households of Pregnant Women in Kinshasa, Democratic Republic of Congo.

The World Health Organization Africa region has high regional hepatitis B virus (HBV) prevalence, and evidence suggests more frequent horizontal HBV transmission than other regions. Context-specific epidemiological studies are needed to inform additional HBV prevention measures. In the cross-sectional Horizontal and Vertical Transmission of Hepatitis B (HOVER-HBV) study, we introduced HBV surface antigen (HBsAg) screening alongside existing HIV screening as part of routine antenatal care in high-volume maternity clinics in Kinshasa, Democratic Republic of Congo. We recruited households of pregnant women ("index mothers") who were HBsAg-positive and HBsAg-negative, defining households as index-positive and index-negative, respectively. Household members underwent HBsAg testing and an epidemiological survey. We evaluated HBsAg prevalence and potential transmission correlates. We enrolled 1006 participants from 200 households (100 index-positive, 100 index-negative) across Kinshasa. HBsAg-positivity prevalence was more than twice as high in index-positive households (5.0% [95% confidence interval {CI}, 2.8%-7.1%]) as in index-negative households (1.9% [95% CI, .6%-3.2%]). HBsAg-positivity prevalence was 3.3 (95% CI, .9-11.8) times as high among direct offspring in index-positive versus index-negative households. Factors associated with HBsAg positivity included older age, marriage, and having multiple recent partners or any new sexual partners among index mothers; and older age, lower household wealth, sharing nail clippers, and using street salons among offspring in index-positive households. Vertical and horizontal HBV transmission within households is ongoing in Kinshasa. Factors associated with infection reveal opportunities for HBV prevention efforts, including perinatal prevention, protection during sexual contact, and sanitation of shared personal items.

Cell-free DNA from nail clippings as source of normal control for genomic studies in hematologic malignancies.

Comprehensive genomic sequencing is becoming a critical component in the assessment of hematologic malignancies, with broad implications for patients' management. In this context, unequivocally discriminating somatic from germline events is challenging but greatly facilitated by matched analysis of tumor:normal pairs of samples. In contrast to solid tumors, in hematologic malignancies conventional sources of normal control material (peripheral blood, buccal swabs, saliva) could be highly involved by the neoplastic process, rendering them unsuitable. In this work we describe our real-world experience using cell-free DNA (cfDNA) isolated from nail clippings as an alternate source of normal control material, through the dedicated review of 2,610 tumor:nail pairs comprehensively sequenced by MSK-IMPACT-heme. Overall, we found that nail cfDNA is a robust germline control for paired genomic studies. In a subset of patients, nail DNA may be contaminated by tumor DNA, reflecting unique attributes of the hematologic disease and transplant history. Contamination is generally low level, but significantly more common among patients with myeloid neoplasms (20.5%; 304/1,482) than among those with lymphoid diseases (5.4%; 61/1,128) and particularly enriched in myeloproliferative neoplasms with marked myelofibrosis. When identified in patients with lymphoid and plasma-cell neoplasms, mutations commonly reflected a myeloid profile and correlated with a concurrent/evolving clonal myeloid neoplasm. Donor DNA was identified in 22% (11/50) of nails collected after allogeneic stem-cell transplantation. In this cohort, an association with a recent history of graft-versus-host disease was identified. These findings should be considered as a potential limitation to the use of nails as a source of normal control DNA but could also provide important diagnostic information regarding the disease process.

Nails as optimal source of DNA for molecular identification of 5 decomposed bodies recovered from seawater: from Y-ancestry to personal identification.

Molecular identification of extremely compromised human remains in forensic field is usually performed from DNA typing of bones, which are a difficult sample to work with. Moreover, autosomal STR profiles do not always result in the identification of the donor due to lack of comparisons or non-hit throughout database searching. An attempt to overcome these issues is represented by fingernails as an alternative DNA source and Y-STRs typing to infer both geographical and familial ancestry of the unknown donor. In this study, we analyzed both 24 autosomal and 27 Y-chromosome STRs from unidentified human remains (UHRs) of five males recovered from the water near the southwestern coast of Sardinia by the Italian Harbor Master's Office. Nail clippings provided an optimal source of autologous DNA for molecular identification in a very short time, producing complete autosomal and Y-STR profiles even under conditions of high body degradation. Unfortunately, no match neither compatibility occurred using autosomal STRs (aSTRs), initially. Upon analyzing the Y-haplotypes, we found out they had already been observed in northern Africa, providing us important investigative leads. This prompted the International Criminal Police Organization (INTERPOL) to provide us with references of alleged relatives that were then confirmed to be related. The use of fingernails represents an excellent DNA source especially for genetic identification of decomposed bodies recovered in seawater environment. Notably, DNA extracted from nails gave high-quality Y-STR haplotypes by which predicting paternal ancestry of the unidentified donors may result fundamental in the forensic investigative context.

Micro-distribution of arsenic in toenail clippings using laser ablation inductively coupled plasma mass spectrometry: implications for biomonitoring.

Toenails are a common monitoring tool for arsenic exposure, but the risk of external contamination of toenails has cast doubt on its usefulness. The main objective of this study is to investigate the micro-distribution of arsenic through the dorsoventral plane of nail clippings to understand endogenous vs exogenous sources. We used laser-ablation inductively coupled plasma mass spectrometry to measure arsenic through a dorsoventral cross-section of the nail plate collected from reference (N = 17) and exposed individuals (N = 35). Our main results showed (1) bulk toenail concentrations measured using ICP-MS in this study ranged from 0.54 to 4.35µg/g; (2) there was a double-hump pattern in arsenic concentrations, i.e., dorsal and ventral layers had higher arsenic than the inner layer; (3) the double-hump was more pronounced in the exposed group (ventral: 6.25μg/g; inner: 0.75μg/g; dorsal: 0.95μg/g) than the reference group (ventral: 0.58μg/g; inner: 0.15μg/g; dorsal: 0.29μg/g) on average; (4) the distribution was, in part, associated with different binding affinity of nail layers (i.e., ventral > dorsal > inner); (5) most individuals in the higher exposure group showed > 25% contamination in ventral and dorsal nail layers; and (6) there were no statistically significant correlations between LA-ICP-MS arsenic with either bulk toenail arsenic or urine arsenic from the same individuals. Our results on micro-distribution and binding affinity provide insight into the impact of external contamination on arsenic concentrations and show how LA-ICP-MS can access the protected inner nail layer to provide a more accurate result.

New Formulation-Microporation Combination Approaches to Delivering Ciclopirox across Human Nails.

Topical treatments for onychomycosis are of interest to those seeking to avoid systemic drug interactions and to improve systemic safety. This work aimed to develop aqueous-based, simple, and cost-effective vehicles that provide high solubility for ciclopirox and enable the delivery of an active through channels created by nail microporation. Following solubility tests, aqueous gels and thermogels based on hydroxypropylmethylcellulose and poloxamer 407, respectively, were loaded with 8% and 16% ciclopirox. Their performance was then compared to the marketed lacquer Micolamina® in in vitro release tests with artificial membranes and in in vitro permeation tests with human nail clippings with and without poration. Finally, a microbiological assay compared the best gel formulations and the reference product. Little correlation was observed between the in vitro release and the permeation data, and the drug release was highly membrane-dependent. Ciclopirox nail retention in single-dose, porated nails tests was larger than in daily-dosing, non-porated nail conditions. The series of new gel and thermogel vehicles delivered ciclopirox more effectively than Micolamina® in single-dose, porated nail experiments. The inhibition of Trichophyton rubrum activity was significantly increased with microporated nails when the gel formulations were applied but not with Micolamina®. Overall, the results suggest that the new vehicles could be successfully combined with nail microporation to improve the drug delivery and efficacy of topical antifungal medication while reducing the dosing frequency, facilitating patients' adherence.

Development of microemulgel formulations with varied permeation enhancers for transungual delivery of luliconazole in onychomycosis management

Luliconazole-loaded microemulgels containing different permeation enhancers were formulated for transungual drug delivery for the management of onychomycosis, onychomycosis, which affects nails. The physicochemical properties like droplet size, zeta potential, pH, viscosity, spreadability, extrudability, oil binding capacity, drug content, and microscopic study were evaluated. The Pseudo-ternary phase diagram was constructed for the formulation of microemulsions (MEs) by keeping the Km ratio constant at 3:1 and characterized for clarity, mean droplet size, zeta potential, viscosity, pH, transmittance, refractive index, and stability. The ME mean droplet size and zeta potential were found in the range of 38.78 to 171.4 nm, and 0.00 to − 6.6 mV, respectively. Prepared MEs were converted into microemulgel by adding a 2.5% gelling agent (Carbapol 934) in the external phase, and a drug release study was conducted. Formulation E3 showed better drug release and was chosen as the control. Four different penetration enhancers were added separately within E3 and further evaluated for pH, viscosity, spreadability, extrudability, oil binding capacity, drug content, microscopic study, Compatibility study, XRD, and DSC. A favorable docking score was observed between luliconazole and Lanosterol 14-alpha-demethylase. In-vitro cumulative drug release at the end of 24 h from E3-SS, containing sodium sulfide as a penetration enhancer, was found to be 94.70% and was 2 times more than the control formulation. Ex-vivo transungual permeation studies through cutting nail clippings were found to be in the range of 28.18 - 36.52 µg/mm2. The microemulgels tagged as E3, E3-SS, and E3-SL showed a significant zone of inhibition against Candida albicans and Aspergillus fumigatus as compared to the marketed formulation.

Detection of 3-4 Methylenedioxyamphetamine from Drug Abuser’s Fingers and Toenails using Liquid Chromatography with Mass Spectroscopy

information regarding drug abuse and pharmaceutical use. In recent years, drug analysis in human nail clippings has proven its significant value in forensic toxicological applications, identification of in utero drug exposure, monitoring of drug treatment programmes, and therapeutic drug monitoring. Nails have various advantages over conventional matrices (blood and urine), which include a longer detection window (months to years), non-invasive sample collection, and easy storage and transportation. These aspects make nails a very significant matrix for forensic toxicology and therapeutic drug monitoring. Because of the low concentrations of drugs of abuse and pharmaceuticals present in nails and the complexity of the keratinized matrix, analytical techniques need to be more sensitive, and sample preparation is crucial. The aim of the present study is to develop a simple, high-performance liquid chromatography-mass spectroscopy (LC-MS) method for the identification and quantitation of 3,4-methylenedioxyamphetamine (MDA) in fingernail and toenail clippings. Finger and toenail clippings were collected from six users undergoing treatment at a rehab center in Ujjain, M.P., India. Nail clippings were initially decontaminated, then hydrolyzed in 1 M NaOH at 370°C, extracted with ethyl acetate, diluted with methanol, and then subjected to LC-MS analysis. The calibration curve was constructed over the 0.5 to 30 ng/mL concentration range using the MDA reference standard. The limit of detection was calculated at 1.10 ng/mL and the limit of quantification was recorded at 3.67 ng/mL in standard solutions, whereas the respective values in spiked nail clippings were 1.21 and 4.6 ng/mg. The developed method has obtained significant results in original nail clippings with mean concentration ranges of 0.12 ng/mg in fingernails and 0.08 ng/mg in toenails in six abuser samples. The new method developed has been found to be capable of detecting the 3,4-methylendioxyamphetamine MDA drug in nail clippings even after 90 days of drug intake.

Testing for clomiphene in keratinous matrices using LC–MS/MS in doping purpose: Is a single intake of clomiphene detectable in hair and nail clippings?

Clomiphene is a selective estrogen receptor modulator. It is indicated for the treatment of female infertility issues but in sport, it can be misused to stimulate endogenous testosterone secretion in men. Therefore, it has been prohibited at all times by the World Anti-doping Agency. The aim of this study was to get data to be able to interpret concentrations in athletes. A healthy volunteer (male, 62 years-old) ingested a single therapeutic dose of clomiphene (Clomid™, 50 mg). Strands of hair (blond, 4 cm) were collected one month after the ingestion. Body hair (beard, axillary, pubic and chest hair), and finger and toenails were collected over 4–5 months.A previous method was modified to identify and quantify clomiphene in keratinous matrices. 30 mg of specimen were sonicated and incubated in 1 mL of methanol, in presence of 200 pg of clomiphene-D5 (internal standard). After centrifugation and evaporation of the organic phase, the samples were analyzed using LC-MS/MS. Linearity was verified in hair and nail clippings between 1 and 500 pg/mg. The limits of detection and quantification were determined at 0.3 and 1 pg/mg respectively.The study demonstrated that clomiphene tested positive in all the analyzed specimens at 9 pg/mg in head hair, from 28 to 486 pg/mg (body hair) and from 4 to 57 pg/mg (nails).Clomiphene was identified for the first time in multiple keratinous matrices. This study demonstrated that a single oral therapeutic dose is detectable in keratinous matrices over a long period of time.

Reduced Level of Prolylhydroxyproline in the Nail Clippings of Oral Cancer Patients and its Role as an Activator of Phospholipase C-β2.

The oral cancer microenvironment plays an important role in the development and progression of the disease which depicts the heterogeneous nature of diseases. Several cellular and non-cellular factors, including dipeptides, have been reported to drive tumor progression and metastasis. Among various secreted molecules in the tumor microenvironment, prolylhydroxyproline (Pro-Hyp) is a collagen-degraded product with specific relevance to fibrosis and oral cancer. However, the detection of Pro-Hyp in the nails of oral cancer patients is a potential biomarker, and our understanding of the biological relevance of Pro-Hyp is highly limited. Here, the authors have attempted to use a novel and in-house vertical tube gel electrophoresis (VTGE) protocol to evaluate the level of Pro-Hyp in the nails of oral cancer patients and healthy subjects. Furthermore, we employed molecular docking and molecular dynamics (MD) simulations to predict the biological function of Pro-Hyp. ADME profiles such as the druglikeness and leadlikeness of Pro-Hyp and a known PLC-β2 activator, m-3M3FBS, were evaluated by the SWISS-ADME server. We report that among various key metabolites, Pro-Hyp, a dipeptide, is reduced in the nails of oral cancer patients. Molecular docking and MD simulations helped to suggest the potential role of Pro-Hyp as an activator of Phospholipase C-β2 (PLC-β2). Pro-Hyp displayed good binding affinity (-7.6 kcal/mol) with specific interactions by a conventional hydrogen bond with key residues, such as HIS311, HIS312, VAL641, and GLU743. MD simulations showed that the activator binding residues and stability of complexes are similar to the well-known activator m-3M3FBS of PLC-β2. ADME profiles such as the druglikeness and leadlikeness of Pro-Hyp were found to be highly comparable and even better than those of m-3M3FBS. This study is one of the first reports on Pro-Hyp as a metabolite biomarker in the nails of oral cancer patients. Furthermore, the implications of Pro-Hyp are proposed to activate PLC-β2 as a pro-tumor signaling cascade. In the future, diagnostic and therapeutic approaches may be explored as biomarkers and mimetic of Pro-Hyp.

Effect existence of aging on stutter ratio evaluated via Bayesian inference

The stochastic behavior of the stutter ratio (SR) in capillary electrophoresis-based DNA typing is currently described and predicted using statistical models in forensic genetics. Clarifying this behavior can help obtain more objective and robust evidence to the court in terms of mixture interpretation. This study aimed to investigate the effect existence of aging on SR via a Bayesian framework. Nail scrapings and clippings were collected from 68 healthy individuals with informed consent. Samples were classified by age-class: young group (0–16 years; n = 36) and older-adult group (>61 years; n = 32). Then, they were compared in terms of their SRs for each simple repeat locus included in GlobalFiler Kit. Bayesian modeling was performed with lognormal distribution model, which implemented multiple linear regression, allele and age-class as explanatory variables. For all simple repeat loci, the median of the posterior distribution of the age-class parameter was a positive value. For CSF1PO and D7S820, the 95% credible interval of the posterior distribution did not include 0. Our data suggested that aging slightly increases the SR. These findings might help elucidate the stochastic behavior of SR.

Testing for clomiphene in keratinous matrices (hair and nail).

Clomiphene or clomifene is a selective estrogen receptor modulator (SERM), used to treat female fertility in case of ovulatory dysfunction. In sport, clomiphene is prohibited at all times for use by athletes and is listed in the section S4.2 (hormone and metabolic modulators) by the World Anti-Doping Agency (WADA). Indeed, clomiphene can indirectly increase testosterone levels in the body and can mitigate some side effects of synthetic steroid abuse. Despite its prescription to millions of subjects, its detection in human hair or nail clippings has never been reported. The aim of this study was to develop a specific method to identify clomiphene in hair and nail clippings by LC-MS-MS. The procedure was then applied in a case of challenged doping results. The method involves sonication/incubation for 1 hour of 30 mg of pulverized material in 1 mL of methanol in presence of 2 ng diazepam-d5 used as internal standard. The chromatographic separation was performed using a HSS C18 column with a 15 min gradient elution. After spiking blank hair and nail with the corresponding amounts of clomiphene, linearity was verified from 1 to 500 pg/mg (r² = 0.9994 and 0.9995 for hair and nail, respectively). The limit of detection was estimated at 0.3 pg/mg for both matrices. No interference was noted from endogenous compounds, particularly steroids. Clomiphene was identified at 85 and 20 pg/mg in the pubic hair and the fingernail clippings, respectively, of a male athlete challenging an adverse analytical finding.

X- ray experimental set-up for in-vitro nail test

In this investigation, the elements Ca and S were evaluated in a single nail clipping using compact and portable X-ray spectrometer model X-123 SDD with Ag target. Experimental conditions for current, voltage and excitation time were investigated. These analyses can be useful for a variety of applications, including nutritional and medical diagnosis, such as, the evaluation of bone dysfunctions by measurement of Ca (bone decalcification) and S for nutrition evaluations (collagen production). This analysis offers some benefits comparatively to blood and serum analyses, such as, non-invasive collection, fast analyses (minutes) and low cost. In addition, nail clippings are simple to obtain, easy to store, and easy to transport. This pilot study show a positive expectation for clinical application using in-vitro nail test.