Developing BIOTEL: A Semi-Automated Spreadsheet for Estimating Telomere Length and Biological Age

Introduction: Telomere length (TL) is causally related to aging and several age-related diseases. Specifically, the abundance of short telomeres and the rate of telomere shortening are strong determinants of cell homeostasis. Thus, tools for analyzing and manipulating TL data can vastly improve research focused on aging. Aim: In this study, we developed a semi-automated worksheet, BIOTEL, to generate individual and group TL statistics and provide a crude estimation of biological age.Results: Data from the Telomere Length Database Project (TLDP) were implemented to the spreadsheet to produce TL statistics. 150 participants were included, and their age was from 21 to 82 years, and the sex distribution ratio was 52.3%: 47.7% (male: female). Initially, we analyzed the fluorescence intensities of telomeres that were measured on metaphase spread leukocytes using three-dimensional (3D) quantitative-fluorescent in situ hybridization (Q-FISH) procedures (3D DNA FISH) with a (C3TA2)3 peptide nucleic acid (PNA) probe. Raw data of fluorescence intensities, demographic data and medical records from the participants were imported into the worksheet. Basic statistical analyses of TL data were provided through BIOTEL, including TL percentiles, specialized charts for TL distribution including the percentage of critically short telomeres (< 3,000 kilobases), individual telomere profiles, and graphs of biological age vs. chronological age.Conclusion: BIOTEL ver. 2.4 is a functional semi-automated worksheet that calculates a wide range of TL statistics, thus a useful tool with applications in research of telomeres and biological age estimation

Biomonitoring of parabens in wild boars through hair samples analysis.

Parabens are compounds widely utilized in the industry as preservative additives to personal care products, cosmetics and food. They pollute the environment and penetrate to the living organisms through the digestive tract, respiratory system and skin. Till now the knowledge about exposure of terrestrial wild mammals to parabens is extremely scarce. Therefore, this study for the first time assessed the concentration levels of five parabens commonly used in industry (methylparaben-MeP, ethylparaben-EtP propylparaben-PrP, benzylparaben -BeP and butylparaben-BuP). Substances have been analyzed in hair samples collected from wild boars using liquid chromatography-mass spectrometry (LC-MS) method. The hair is a matrix, which allows to study long-term exposure of organisms to parabens. During this study MeP was noted in 96.3% of samples with mean 88.3±72.9 pg/mg, PrP in 87.0% of samples with mean 8.5±3.3 pg/mg, BeP in 44.4% of samples with mean 17.2±12.3 pg/mg and EtP in 11.1% of samples with mean 17.2±4.8 pg/mg. In turn BuP was noted only in 3.7% of samples with concentration levels below limit of quantification (2.6 pg/mg). Statistically significant intragender differences in parabens levels have not been noted. Only BeP concentration levels depended on industrialization and density of human population of area, where the animals lived. This study indicates that wild boars are exposed to parabens, especially to MeP and PrP, and analysis of the hair seems to be a useful tool of biomonitoring of parabens in wild mammals

The Presence of Triclosan in Human Hair Samples in Poland—A Pilot Study

Triclosan (TCS) is an organic substance showing antibacterial action, which is commonly used in many branches of industry, including, among others, cosmetics, pharmaceuticals and the food industry. TCS may penetrate into living organisms and negatively affect the health of humans and animals. The majority of previous investigations on TCS biomonitoring in humans have been performed on urine, but currently, studies on hair samples are becoming increasingly important. The aim of this study was to evaluate TCS concentration levels in residents of Olsztyn, a city in northeastern Poland, using a liquid chromatography-mass spectrometry technique. The presence of TCS was observed in 96.7% of samples tested, with concentration levels from 37.9 pg/mg to 3386.5 pg/mg. The mean concentration level of TCS in the present study was 402.6 (±803.6) pg/mg, and the median value was 103.3 pg/mg. Although there were some differences in TCS concentration levels between males and females, humans of various ages and humans with colored and natural hair had no statistically significant differences in TCS concentration levels. The obtained results have clearly indicated that people living in northeastern Poland are exposed to TCS to a large degree, and hair analysis, despite some limitations, is a suitable method for TCS biomonitoring in humans

Достижения в области биомониторинга фосфорорганических и хлорорганических пестицидов

Organophosphorus (OPs) and organochlorine (OCs) pesticides are synthetic organic compounds used mainly in agriculture to manage pest problems. Human is occupationally and environmentally exposed to pesticides via inhalation, ingestion and skin absorption. When the pollutant enters human body it is rapidly metabolized to more toxic compounds than the parent compound. Their ability to interfere with biochemical and metabolic pathways in humans makes them compounds of high concern. Acute and chronic exposure has been linked with neurological, cardiovascular, birth effects and male infertility. Human exposure to pesticides is estimated by measuring their metabolites in biological tissues and fluids such as blood, urine, amniotic fluid and hair. In this review study advances on biomonitoring of pesticides are presented. Measured levels of pesticides in urine matrices, amniotic fluid, meconium and hair are presented in order to monitor the human burden and biomonitoring data between occupationally and environmentally exposed populations are being comparedФосфорорганические и хлорорганические пестициды – это синтетические органические соединения, используемые преимущественно в сельском хозяйстве для борьбы с вредителями. Люди подвергаются воздействию пестицидов в процессе профессиональной деятельности и при контакте с зараженной средой через их вдыхание, потребление с пищей и поглощение кожей. При попадании в организм человека поллютант быстро метаболизируется до более токсичных соединений, чем исходное вещество. Их способность взаимодействовать с биохимическими и метаболическими путями в человеческом организме делает их потенциально опасными соединениями. Обнаружена связь между острым и хроническим воздействием пестицидов и неврологическими и кардиоваскулярными эффектами, врожденными отклонениями и мужским бесплодием. Оценка воздействия пестицидов на организм человека проводится путем измерения их метаболитов в биологических тканях и жидкостях, таких как кровь, моча, амниотическая жидкость, а также в волосах. В данном обзоре представлены достижения в области биомониторинга пестицидов. Для оценки нагрузки на человека приведены измеренные уровни пестицидов в моче, амниотической жидкости, меконии и волосах. Проведено сравнение популяций, подвергшихся воздействию пестицидов в связи с производственной деятельностью и при контакте с окружающей средо

Telomerase inhibitors and activators in aging and cancer : A systematic review

The main aim of the present systematic review was to summarize the most frequently used telomerase regulators with an impact on aging and cancer that are referred to in in vitro and in vivo studies. For this purpose, a systematic review of the available literature on telomerase regulators referred to in articles from PubMed and Scopus libraries published from 2002 to 2021 and in accordance with PRISMA 2020 criteria, was conducted. Articles were included if they met the following criteria: They referred to telomerase modulators in aging and in cancer and were in vitro and/or in vivo studies, while studies that did not provide sufficient data or studies not written in English were excluded. In the present systematic review, 54 publications were included, of which 29 were full-text published studies, 11 were full-text reviews, 10 structure-based design studies and 4 abstracts are reported in this review. Telomerase regulators were then categorized as synthetic direct telomerase inhibitors, synthetic indirect telomerase inhibitors, synthetic telomerase activators, natural direct telomerase activators, natural telomerase inhibitors and natural indirect telomerase activators, according to their origin and their activity. On the whole, as demonstrated herein, telomerase regulators appear to be promising treatment agents in various age-related diseases. However, further in vivo and in vitro studies need to be performed in order to clarify the potentiality of telomerase as a therapeutic target.Title in WoS: tau elomerase inhibitors and activators in aging and cancer: A systematic review</p

Antibiotics in Raw Meat Samples: Estimation of Dietary Exposure and Risk Assessment

The extensive use of antibiotics in livestock farming poses increased concerns for human health as residues of these substances are present in edible tissues. The aim of this study was the determination of the levels of four groups of antibiotics (sulfonamides&mdash;SAs, tetracyclines&mdash;TCs, streptomycines&mdash;STr and quinolones&mdash;QNLs) in meat samples (muscles, livers and kidneys from beef, chicken and pork) and the estimation of the dietary exposure to antibiotics from meat consumption and the potential hazard for human health. Fifty-four samples of raw meat were randomly collected in 2018 from the Cretan market, Greece and analyzed both with an enzyme-linked immunosorbent assay (ELISA) and liquid chromatography&ndash;mass spectrometry (LC&ndash;MS). According to the results derived from the ELISA method, only 2% of the meat samples were free from antibiotics, 2% were detected with 4 antibiotics and the great majority of the samples (87%) were detected with 2 to 3 antibiotics. SAs presented the highest detection frequencies for all samples whereas TCs were not detected in any bovine sample. The highest median concentration was detected for STr in bovine muscles (182.10 &mu;g/kg) followed by QNLs (93.36 &mu;g/kg) in pork kidneys whereas the chicken samples had higher burdens of QNLs compared to the other meat samples. LC&ndash;MS analysis showed that oxytetracycline (OTC) was the most common antibiotic in all samples. The highest median concentration of all antibiotics was detected for doxycycline (DOX) (181.73 &mu;g/kg in pork kidney) followed by OTC in bovine liver (74.46 &mu;g/kg). Risk characterization was applied for each of the two methods; The hazard quotients (HQ) did not exceed 0.059 for the ELISA method and 0.113 for the LC&ndash;MS method for any group of antibiotics, whereas the total hazard indexes (HI) were 0.078 and 0.021, respectively. The results showed the presence of different groups of antibiotics in meat from the Cretan market and that the health risk to antibiotics is low. A risk assessment analysis conducted for meat consumption and corrected for the aggregated exposure revealed no risk for the consumers

Достижения в области биомониторинга фосфорорганических и хлорорганических пестицидов

Organophosphorus (OPs) and organochlorine (OCs) pesticides are synthetic organic compounds used mainly in agriculture to manage pest problems. Human is occupationally and environmentally exposed to pesticides via inhalation, ingestion and skin absorption. When the pollutant enters human body it is rapidly metabolized to more toxic compounds than the parent compound. Their ability to interfere with biochemical and metabolic pathways in humans makes them compounds of high concern. Acute and chronic exposure has been linked with neurological, cardiovascular, birth effects and male infertility. Human exposure to pesticides is estimated by measuring their metabolites in biological tissues and fluids such as blood, urine, amniotic fluid and hair. In this review study advances on biomonitoring of pesticides are presented. Measured levels of pesticides in urine matrices, amniotic fluid, meconium and hair are presented in order to monitor the human burden and biomonitoring data between occupationally and environmentally exposed populations are being comparedФосфорорганические и хлорорганические пестициды – это синтетические органические соединения, используемые преимущественно в сельском хозяйстве для борьбы с вредителями. Люди подвергаются воздействию пестицидов в процессе профессиональной деятельности и при контакте с зараженной средой через их вдыхание, потребление с пищей и поглощение кожей. При попадании в организм человека поллютант быстро метаболизируется до более токсичных соединений, чем исходное вещество. Их способность взаимодействовать с биохимическими и метаболическими путями в человеческом организме делает их потенциально опасными соединениями. Обнаружена связь между острым и хроническим воздействием пестицидов и неврологическими и кардиоваскулярными эффектами, врожденными отклонениями и мужским бесплодием. Оценка воздействия пестицидов на организм человека проводится путем измерения их метаболитов в биологических тканях и жидкостях, таких как кровь, моча, амниотическая жидкость, а также в волосах. В данном обзоре представлены достижения в области биомониторинга пестицидов. Для оценки нагрузки на человека приведены измеренные уровни пестицидов в моче, амниотической жидкости, меконии и волосах. Проведено сравнение популяций, подвергшихся воздействию пестицидов в связи с производственной деятельностью и при контакте с окружающей средо

Long-term exposure of rabbits to imidaclorpid as quantified in blood induces genotoxic effect

The present in-vivo study focuses on the genotoxic effect of the neonicotinoid pesticide imidacloprid (IMI) in rabbits. The purpose of the study was to establish a possible relationship between exposure to the pesticide (dose and duration) and genotoxicity. Furthermore, an analytical method for the simultaneous determination of IMI and its major metabolite 6-chloronicotinic acid (6-ClNA) in blood was developed and validated. The isolation of the two analytes from blood was performed by liquid-liquid extraction with dichloromethane. Analysis was performed by Liquid Chromatography - Atmospheric Pressure Chemical Ionization - Mass Spectrometry (LC-APCI-MS). The method was applied on the determination of IMI and 6-ClNA in serum samples obtained from rabbits fed with the insecticide at two low doses. Furthermore, parameters of genotoxicity and cytotoxicity were evaluated by measuring binucleated cells with micronuclei (BNMN), micronuclei (MN) and the Cytokinesis Block Proliferation Index (CBPI), in lymphocytes of exposed rabbits. The results revealed a genotoxic effect of IMI for both exposed groups. There were statistically significant differences in the frequencies of BNMN and MN between control and exposed groups but there was no dose-dependence, neither time-dependence of the genotoxic effect for the administered doses. This is the first time that long term exposure to IMI in rabbits was studied for the determination of its genotoxic effect. The genotoxic effect of IMI as it is depicted by the current study is in accordance with previous studies. (C) 2016 Published by Elsevier Ltd

DNA Damage Estimation after Chronic and Combined Exposure to Endocrine Disruptors: An In Vivo Real-Life Risk Simulation Approach

Exposure to chemical substances has always been a matter of concern for the scientific community. During the last few years, researchers have been focusing on studying the effects resulting from combined exposure to different substances. In this study, we aimed to determine the DNA damage caused after chronic and combined exposure to substances characterized as endocrine disruptors using comet and micronuclei assays, specifically glyphosate (pure and commercial form), bisphenol A, parabens (methyl-, propyl- and butylparaben), triclosan and bis (2-ethylhexyl) phthalate. The highest mean tail intensity was observed in the group exposed to a high-dose (10 × ADI) mixture of substances (Group 3), with a mean value of 11.97 (11.26–13.90), while statistically significant differences were noticed between the groups exposed to low-dose (1 × ADI) (Group 2) and high-dose (10 × ADI) (Group 3) mixtures of substances (p = 0.003), and between Group 3 and both groups exposed to high doses (10 × ADI) of the pure and commercial forms of glyphosate (Groups 4 (p = 0.014) and 5 (p = 0.007)). The micronuclei assay results were moderately correlated with the exposure period. Group 5 was the most impacted exposure group at all sampling times, with mean MN counts ranging between 28.75 ± 1.71 and 60.75 ± 1.71, followed by Group 3 (18.25 ± 1.50–45.75 ± 1.71), showing that commercial forms of glyphosate additives as well as mixtures of endocrine disruptors can enhance MN formation. All exposure groups showed statistically significant differences in micronuclei counts with an increasing time trend

In Vivo Estimation of the Biological Effects of Endocrine Disruptors in Rabbits after Combined and Long-Term Exposure: Study Protocol

Recently, an increasing number of chemical compounds are being characterized as endocrine disruptors since they have been proven to interact with the endocrine system, which plays a crucial role in the maintenance of homeostasis. Glyphosate is the active substance of the herbicide Roundup&reg;, bisphenol A (BPA) and di (2-ethylhexyl) phthalate (DEHP) are used as plasticizers, while triclosan (TCS), methyl (MePB), propyl (PrPB), and butyl (BuPB) parabens are used as antimicrobial agents and preservatives mainly in personal care products. Studies indicate that exposure to these substances can affect humans causing developmental problems and problems in the endocrine, reproductive, nervous, immune, and respiratory systems. Although there are copious studies related to these substances, there are few in vivo studies related to combined exposure to these endocrine disruptors. The aim of the present pilot study is the investigation and assessment of the above substances&rsquo; toxicity in rabbits after twelve months of exposure to glyphosate (both pure and commercial form) and to a mixture of all the above substances at subtoxic levels. The lack of data from the literature concerning rabbits&rsquo; exposure to these substances and the restrictions of the 3Rs Principle will result in a limited number of animals available for use (four animals per group, twenty animals in total)