Structural Chromosome Aberrations Research Articles

Toxicity Profile Study of Antihypertensive Drug Prazosin in Pregnant Wistar Rats

The purpose of this research is to investigate and assess the cytotoxicity and genotoxicity of Prazosin HCL in pregnant rats. Prazosin (PZ) was administered to the animals intraperitoneally (IP) at dosages of 5, 15, and 25 mg/kg/body weight for single dose (14-day) toxicity tests. The following parameters have been examined for evaluating genotoxicity: bone marrow micronucleus assay, peripheral blood micronucleus assay, DNA damage is measured using the metaphase chromosomal analysis, DNA damage is measured using the DNA fragmentation test, and cytotoxicity is measured using a histological analysis. The results obtained clearly demonstrate that PZ induced hazardous responses at the higher dose in the hepatocytes, as evidenced by DNA damage, and increased DNA fragmentation in pregnant rats. Observing that PZ significantly increased DNA strand breakage and structural chromosomal aberrations in bone marrow cell lines is also fascinating. As a result, it is thought to be genotoxic to bone marrow and mouse hepatocyte cells. The current study showed that Prazosin had significant genotoxic effects in pregnant rats at the matching hepatotoxic dose level.

Detection of chromosome aberrations in 17 054 individuals with fertility problems and their subsequent assisted reproductive technology treatments in Central China.

How do the types and frequency of chromosome aberrations in couples in central China affect fertility and ART treatment? Men with chromosome aberrations or polymorphisms have an increased risk of semen quality impairment and infertility, and couples affected by reciprocal translocations had a lower pregnancy rate compared with other chromosome aberrations. Karyotyping is crucial for patients affected by infertility as chromosome aberrations play an important role in the etiology of male infertility. However, the influence of chromosome aberrations and polymorphisms on sperm motility and morphology remains controversial. Data on ART treatment outcomes in infertile couples affected by chromosome aberrations are insufficient. We conducted a retrospective study involving 17 054 patients affected by infertility who underwent karyotyping in our center between January 2020 and May 2022. Karyotyping was performed on 17 054 patients with reproductive failure. All patients were from the central regions of China. The following data were collected from a medical records system using patient identification numbers: couples' ages, history of pregnancy and childbirth, type of infertility, years of infertility, cause of infertility, chromosome karyotypes, semen analysis results, assisted reproductive techniques performed, and treatment outcomes of ART. The incidence of chromosome aberrations was 2.04%; 2.49% in men and 1.57% in women. By analyzing the relationships between chromosome aberrations/polymorphisms and abnormal semen parameters, we found that there were significantly higher rates of asthenozoospermia, oligospermia, and teratozoospermia among men with Robertsonian translocations and sex chromosomal structural aberrations compared with those with normal karyotypes. Higher rates of asthenozoospermia and teratozoospermia were also observed among men with autosomal reciprocal translocations. The incidence of chromosome aberrations in azoospermic men (13.75%), and in men with cryptozoospermia or severe oligospermia (6.97%) was significantly higher than that in men with mild oligospermia or normospermia (0.88-2.12%). In addition, we found that the progressive movement of sperm is impaired in men with Chromosome 21 polymorphisms compared with men with normal karyotypes (39.46% ± 20.51% vs 48.61% ± 18.76%, P = 0.026). The percentage of morphologically normal forms was lower in the chromosomal polymorphism group than in the normal karyotype group (5.01% ± 2.41% vs 5.59% ± 2.14%, P = 0.001), especially in men with polymorphisms on Chromosome 9 (enlarged Chromosome 9 heterochromatin [9qh+]: 4.48% ± 2.22% vs 5.59% ± 2.14%, P = 0.006; pericentric inversion of Chromosome 9 [inv(9)]: 5.09% ± 3.11% vs 5.59% ± 2.14%, P = 0.008). ART treatment was successful in 36.00% of couples affected by chromosome aberrations. However, couples affected by reciprocal translocations achieved a lower pregnancy rate (24.07%), which may be due to the lower euploidy rates (27.31%) when compared with that in other chromosome aberrations. First, although the initial cohort was large, chromosome aberrations were identified in a small number of patients. Second, the observational nature of the study design is limiting. Third, the couples affected by infertility in this study were all outpatients that did not undergo identical comprehensive examinations except for karyotyping, leading to the incomplete collection of medical records. Also, the population included in this study mainly focused on couples affected by infertility, which may not be included in the European Association of Urology (EAU) recommendation on male infertility. Men with chromosome aberrations or polymorphisms have an increased risk of semen quality impairment and infertility. Constitutional chromosome analysis is recommended for men affected by infertility and severe oligospermia or azoospermia to facilitate early and appropriate guidance for the most suitable treatment. Carriers of chromosome aberrations can achieve acceptable pregnancy outcomes through IVF. However, couples affected by reciprocal translocations have lower pregnancy rates, and more treatment cycles are needed before a successful pregnancy. A possible explanation may be the fewer euploid embryos obtained. This work was supported by Grant 2021YFC2700603 from the National Key Research & Development Program of China. The authors declare no conflicts of interest. N/A.

Preferential Efficacy of Remission Maintenance with HDC/IL-2 in Age Groups of Patients with Chemoresponsive or Normal Karyotype AML

Background Relapse in complete remission (CR) remains a leading cause of morbidity and mortality in acute myeloid leukemia (AML). Previous phase 3 trial data show that combinatorial immunotherapy with histamine dihydrochloride and low-dose interleukin-2 (HDC/IL-2) reduces relapse incidence in the post-consolidation phase, in particular among younger adult AML patients (<60 years old) in CR1 (Brune et al., Blood 108:88). Recent post-hoc analyses identified subgroups of responding patients including (1) those achieving CR1 after the first induction course of chemotherapy (referred to as chemoresponsive AML) and (2) those carrying leukemic cells of normal karyotype (NK-AML), i.e. devoid of structural or numerical chromosomal aberrations. Here we report analyses of treatment efficacy in a phase 3 trial within age groups of patients with chemoresponsive AML or NK-AML. Methods Three hundred and twenty AML patients (18-84 years old, median 57) who were ineligible for allogeneic stem cell transplantation were randomly assigned to receive HDC/IL-2 or no treatment (control) for remission maintenance in a phase 3 trial. Patients were recruited in Europe, Israel, Oceania, and USA. Treatment was initiated in CR after the completion of consolidation chemotherapy and comprised 10 consecutive 3-week cycles of HDC/IL-2 for 18 months (mo) with 3- or 6-week rest periods. In each cycle, patients received HDC at 0.5 mg and human recombinant low-dose IL-2 (aldesleukin; 16,400 IU/kg) s.c. twice daily (n=160) or no treatment (control; n=160). After 18 months of treatment, all patients were monitored for >18 additional mo (median follow-up 48 mo). Results Among study subjects in CR1 203/261 (78%) of evaluable patients attained CR after the first induction cycle (chemoresponsive AML) and 128/220 (58%) had NK-AML. Within both subgroups, analysis of efficacy by age at diagnosis implied preferred benefit of HDC/IL-2 vs. control in patients 40-59 years old (Figure 1) with independent efficacy in patients 40-49 and 50-59 years old (Table 1). The treatment arm was not significantly superior over the control arm for LFS or OS in the youngest patients (18-39 years old) or in patients >60 years old (P>0.2, logrank test, for all comparisons (LFS and OS)) within subgroups of chemosensitive AML or NK-AML. Conclusions We speculate that the observed benefit of HDC/IL-2 in patients responding swiftly to initial chemotherapy may be explained by more efficient immune-mediated elimination of leukemic cells at a lower burden of leukemia. The preferential efficacy of HDC/IL-2 in NK-AML points towards efficacy in subgroups of leukemic mutations that are less common in the youngest group of patients. The reason for the lack of significant efficacy in patients >60 years old, despite achievement of CR1 after the initial chemotherapy and despite having NK-AML, remains unknown but may be explained by fading anti-leukemic immunity in old patients. Overall, these findings may be considered for the selection of patients suitable for immunotherapy with HDC/IL-2 in an emerging landscape of strategies for relapse prevention in AML.

Protective Effect of Cafestol on Doxorubicin-induced Genotoxicity in Rats

Doxorubicin is an efficient antineoplastic agent that has a broad antitumour spectrum; however, its genotoxic adverse effects on normal cells can be produced through oxidative damage, and this limits its clinical application. Cafestol is a naturally-occurring diterpene in unfiltered coffee with noteworthy antioxidant, antimutagenic and anti-inflammatory activities.
 The present study aimed to investigate the possible protective effect of cafestol against doxorubicin-induced chromosomal and DNA damage in rat bone marrow cells. Wistar
 Albino rats of both sexes were administered cafestol (5mg/kg body weight once daily for 14 consecutive days) by oral gavage alone or with doxorubicin which was injected as a single dose (90 mg/kg intraperitoneally at day 14) to induce toxicity. The bone marrow was harvested 24 hours after doxorubicin’s injection in all groups for the assessment of structural chromosomal aberration, micronucleus, and comet assays. The result showed that rats in the doxorubicin-only group exhibited a significant decrease (P<0.05) in mitotic index with a significant elevation (P<0.05) in the % DNA in Tail, micronucleus appearance and total structural chromosomal aberrations compared to those of the negative control group; while oral administration of cafestol 14 days prior to doxorubicin, significantly-reduced the % DNA in Tail, micronucleus appearance, and the total number of chromosomal aberrations (P<0.05), and improved the mitotic index compared to rats intraperitoneally-injected with doxorubicin alone. 
 This study revealed that cafestol has protective effects against the genotoxicity induced by doxorubicin.

8q Gain Has No Additional Predictive Value in SF3B1MUT Uveal Melanoma but Is Predictive for a Worse Prognosis in Patients with BAP1MUT Uveal Melanoma

Gain of chromosome 8q has been associated with poor prognosis in uveal melanoma (UM), and an increase in the absolute number of 8q-copies correlated with an even shorter survival. Splicing factor 3b subunit 1 (SF3B1)-mutated (SF3B1MUT) tumors display structural chromosomal anomalies and frequently show a partial gain of chromosome 8qter. A recent subset of SF3B1MUT UM with early-onset metastases has been identified, prompting the investigation of the relationship between survival, 8q gain, and SF3B1MUT UM. Retrospective cohort study. Patients diagnosed with UM who underwent enucleation or received a biopsy at the Erasmus MC Cancer Institute or the Rotterdam Eye Hospital, The Netherlands were included. Fifty-nine patients with SF3B1MUT tumors and 211 patients with BRCA1 associated protein 1 (BAP1)-mutated (BAP1MUT) tumors were included in this study. Copy number status and gene expression were assessed using either a single nucleotide polymorphism array, fluorescence in situ hybridization, and karyotyping, or a combination of these techniques. Disease-free survival was determined and a cut-off of 60 months was used to define early-onset metastatic disease. Disease-free survival. Forty-eight patients with SF3B1MUT UM (81%) had chromosome 8q gain (3 copies, 78%; 4 copies, 22%). Kaplan-Meier analysis of SF3B1MUT UM did not indicate a difference in survival in patients with or without gain of 8q (P= 0.99). Furthermore, the number of 8q copies was not associated with survival when comparing early (P= 0.97) versus late (P= 0.23) metastases group. In contrast, the presence of 8q gain (86%) was correlated with a decreased survival in BAP1MUT UM (P= 0.013). We did not find a correlation between 8q gain and early-onset metastasis in SF3B1MUT tumors. Gain of 8q has no additional predictive value in SF3B1MUT tumors. In contrast, 8q gain is predictive of a worse prognosis in patients with BAP1MUT tumors. Thus, gain of chromosome 8q has additional predictive value for BAP1MUT tumors, but not for SF3B1MUT tumors. The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Cytogenetic damage by vanadium(IV) and vanadium(III) on the bone marrow of mice

Vanadium is a strategic metal that has many important industrial applications and is generated by the use of burning fossil fuels, which inevitably leads to their release into the environment, mainly in the form of oxides. The wastes generated by their use represent a major health hazard. Furthermore, it has attracted attention because several genotoxicity studies have shown that some vanadium compounds can affect DNA; among the most studied compounds is vanadium pentoxide, but studies in vivo with oxidation states IV and III are scarce and controversial. In this study, the genotoxic and cytotoxic potential of vanadium oxides was investigated in mouse bone marrow cells using structural chromosomal aberration (SCA) and mitotic index (MI) test systems. Three groups were administered vanadium(IV) tetraoxide (V2O4) intraperitoneally at 4.7, 9.4 or 18.8 mg/kg, and three groups were administered vanadium(III) trioxide (V2O3) at 4.22, 8.46 or 16.93 mg/kg body weight. The control group was treated with sterile water, and the positive control group was treated with cadmium(II) chloride (CdCl2). After 24 h, all doses of vanadium compounds increased the percentage of cells with SCA and decreased the MI. Our results demonstrated that under the present experimental conditions and doses, treatment with V2O4 and V2O3 induces chromosomal aberrations and alters cell division in the bone marrow of mice.

ОЦЕНКА ГЕНОТОКСИЧЕСКОГО ВОЗДЕЙСТВИЯ МИТОМИЦИНА С НА ПРИМЕРЕ APOE НОКАУТНЫХ МЫШЕЙ

BACKGROUND Mitomycin C (MMC) has a wide spectrum of genotoxicity, including inhibition of DNA synthesis, clastogenesis and mutagenesis. As an immediate-action clastogen requiring exclusively intracellular reductive activation, MMC initiates efficient DNA cross-linking. Most studies are aimed at studying single-stage acute effects, which are caused by high doses of mutagen. In turn, there are no or very few studies aimed at studying the chronic effects of MMC. Polychrome red blood cells have been accepted as a suitable target for the evaluation of micronuclei in both acute and cumulative injury. The in vivo micronucleus test is well established as a standard assay for assessing genotoxicity at the chromosomal level of mouse erythrocytes. THE AIM OF THE STUDY To create a chronic genotoxic effect of mitomycin C without lethal outcome in ApoE knockout mice while selecting the optimal dose of MMC. MATERIALS AND METHODS The study design included 6 groups of ApoE-/- mice, two doses of MMC with a concentration of 0.1 and 0.5 mg/kg, one-time and three-time administration. To assess genotoxicity, 1000 polychrome erythrocytes (PCEs) extracted from the bone marrow of a mousefemur were counted on each sample, and PCEs with micronuclei were identified. RESULTS This study aimed to find the optimal dose of MMC that has a clear genotoxic effect and does not lead to death in an ApoE knockout mouse model. With a single injection, PCEs with chromosomal damage were more common (more than 2 times; p < 0.05) in groups of mice with MMC administration (0.1 and 0.5 mg/kg; 0.39 % and 0.26 %, respectively) compared to the control group (0.15 %). We also found that the frequency of occurrence of PCEs with micronuclei in groups of mice with MMC dose of 0.1 and 0.5 mg/kg (0.36 % and 0.47 %, respectively) and three-time administration exceeded this indicator in mice from the control group (0.2 %). CONCLUSION The present study on the determination of the optimal dose of mitomycin C provides further evidence that 0.1 mg/kg is the threshold value for genotoxic effects caused by MMC. An increase in the frequency of micronucleated immature red blood cells in animals exposed to a mutagen is an indicator of induced structural or numerical chromosomal aberrations. Our results further suggest that careful selection of MMC dose is critical. Dose-response studies in rodents can provide useful information on mechanisms and dose selection for long-term toxicity studies.

Use of Atomic Force Microscopy in UVB-Induced Chromosome Damage Provides Important Bioinformation for Cell Damage Assessment.

The chromosomal structure derived from UVB-stimulated HaCaT cells was detected by atomic force microscopy (AFM) to evaluate the effect of UVB irradiation. The results showed that the higher the UVB irradiation dose, the more the cells that had chromosome aberration. At the same time, different representative types of chromosome structural aberrations were investigated. We also revealed damage to both DNA and cells under the corresponding irradiation doses. It was found that the degree of DNA damage was directly proportional to the irradiation dose. The mechanical properties of cells were also changed after UVB irradiation, suggesting that cells experienced a series of chain reactions from inside to outside after irradiation. The high-resolution imaging of chromosome structures by AFM after UVB irradiation enables us to relate the damage between chromosomes, DNA, and cells caused by UVB irradiation and provides specific information on genetic effects.

Chromosomal aberrations in Ukrainian red-spotted dairy breed cows when using different breeding methods

Aim. The aim of this study was a comparative analysis of karyotypes of purebred, inbred, and crossbred cows of the Ukrainian red-spotted dairy breed in order to detect their degree of chromosomal aberrations. Methods. The cyto- genetic study involved 156 cows of the Ukrainian red-spotted dairy breed from two research farms of M.V. Zubets Institute of Animal Breeding and Genetics, the NAAS. Cultures of peripheral lymphocytes, stimulated with phytohemagglutinin (PHA, Difco, USA) were used according to the method of Moorhead et al. (1960). The chromosomal preparations were stained with 2 % Giemsa solution (Giemsa, Merk) and analyzed under an Axiostar plus (Carl Zeiss, Germany) light microscope at 1,000 times magnification. Metaphase slides were photographed with a digital camera Olympus D-460 ZOOM. The biometric processing of the study results was conducted by ANOVA using Microsoft Excel 2010. Results. Genomic mutations (aneuploidy and polyploidy) and structural aberrations of chromosomes were found in the karyotypes of the investigated purebred, inbred, and crossbred cows. The total share of aberrant cells varied from 11.4 ± 0.39 % in purebred, 11.8 ± 0.40 % in inbred to 17.30 ± 0.85 % in crossbred cows. In two animals the Robertson translocation, rob (1 : 29) was found. A statistically significant difference between purebred, inbred, and crossbred animals was found regarding cells with aneuploidy and structural chromosomal aberrations. Conclusions. The karyological analysis yielded data showing changes in genetic structure that can have an influence on fertility and production in different breeding programs.

Furan promotes cytotoxic effects through DNA damage and cell apoptosis in Leydig cells

Furan is an organic chemical that can cause adverse effects on human health and is formed as a result of the thermal decomposition of many food components during cooking, storage, and processing techniques. Studies have shown that exposure to furan causes nephrotoxicity, hepatotoxicity, immunotoxicity, and reproductive toxicity. According to our current knowledge of the literature, the genotoxic mode of action of furan is highly controversial. The genotoxic effects of furan on the male reproductive system, however, have not been studied. In this study, the TM3 Leydig cell line was treated with 750, 1500, and 3000 μM concentrations of furan for 24 h. Following the completion of the exposure period, the cytotoxicity of furan in TM3 Leydig cells was assessed using a cell viability assay and a spectrophotometric measurement of lactate dehydrogenase (LDH) enzyme levels. The double fluorescence staining method was used to demonstrate furan-induced apoptosis, and DNA damage was shown using the micronucleus, comet, and chromosomal aberration assays. The result indicated that furan administration of Leydig cells resulted in an increase in structural chromosomal aberration, comet, and micronucleus formation, reduced cell viability, increased LDH activity, and a higher incidence of apoptotic cells. These findings revealed that furan induces DNA damage in TM3 Leydig cells, causing genotoxicity and DNA damage-induced cytotoxicity.

Safety assessment of the substance poly(2-hydroxypropanoic acid), n-octyl/n-decyl esters, for use in food contact materials.

The EFSA Panel on Food Contact Materials, Enzymes and Processing Aids (CEP) assessed the safety of poly(2-hydroxypropanoic acid), n-octyl/n-decyl esters (OLA8), which is intended to be used as a plasticiser into polylactic acid (PLA) in contact with non-fatty foods. OLA8 is intended to be used at up to 5% and 15% w/w with or without starch, respectively (or with other additives with similar function). The migration for 10 days at 40°C from the film without starch was 0.16 mg/kg in 10% ethanol and 0.01 mg/kg in 3% acetic acid, while from the film with the starch it was well above 0.05 mg/kg food in all simulants. Some of the testing conditions were inconsistently reported. The substance did not induce gene mutations in bacterial cells and did not induce structural chromosomal aberrations or polyploidy in mammalian cells, thus, does not raise concern for genotoxicity. Instead of providing a 90-day oral toxicity study, a hydrolysis study in ■■■■■ was submitted to read-across from the authorised starting substances, ■■■■■ and the ■■■■■. However, the data provided did not allow to perform the read-across, thus no appropriate toxicological data were provided to support migration above 0.05 mg/kg food (including for contact with 10% ethanol and use in combination with starch). The Panel concluded that OLA8 does not raise a safety concern for the consumer if it is used as an additive at up to 15% w/w in the manufacture of PLA articles that do not contain starch (and other additives with similar function), that are intended to be in contact for 10 days at 40°C with foods simulated by 3% acetic acid and from which the migration does not exceed 0.05 mg/kg food.

Genotoxicity Evaluation of The Novel Psychoactive Substance MTTA.

MTTA, also known as mephtetramine, is a stimulant novel psychoactive substance characterized by a simil-cathinonic structure. To date, little has been studied on its pharmaco-toxicological profile, and its genotoxic potential has never been assessed. In order to fill this gap, the aim of the present work was to evaluate its genotoxicity on TK6 cells in terms of its ability to induce structural and numerical chromosomal aberrations by means of a cytofluorimetric protocol of the "In Vitro Mammalian Cell Micronucleus (MN) test". To consider the in vitro effects of both the parental compound and the related metabolites, TK6 cells were treated with MTTA in the absence or presence of an exogenous metabolic activation system (S9 mix) for a short-term time (3 h) followed by a recovery period (23 h). No statistically significant increase in the MNi frequency was detected. Specifically, in the presence of S9 mix, only a slight increasing trend was observable at all tested concentrations, whereas, without S9 mix, at 75 µM, almost a doubling of the negative control was reached. For the purposes of comprehensive evaluation, a long-term treatment (26 h) was also included. In this case, a statistically significant enhancement in the MNi frequency was observed at 50 µM.

Paternal age, risk of congenital anomalies, and birth outcomes: a population-based cohort study.

The objective of the study was to explore the impact of paternal age on the risk of congenital anomalies and birth outcomes in infants born in the USA between 2016 and 2021. This retrospective cohort study used data from the National Vital Statistics System (NVSS) database, a data set containing information on live birth in the USA between 2016 and 2021. Newborns were divided into four groups based on their paternal age (< 25, 25-34, 35-44, and > 44years) and using the 25-34 age group as a reference. The primary outcomes were congenital anomalies involving structural anomalies and chromosome anomalies. Secondary outcomes were preterm birth, low birth weight, severe neonatal perinatal asphyxia, and admission to neonatal intensive care units (NICU). A multivariable logistic regression model was used to analyze the association between paternal age and outcomes. Overall, 17,764,695 live births were included in the final analyses. After adjusting confounding factors, advanced paternal age > 44years was associated with increased odds of congenital anomalies (adjusted odds ratio (aOR) = 1.17, 95%CI 1.12-1.21) compared with the 25-34 age group, mainly for the chromosomal anomalies (aOR = 1.59, 95%CI 1.40-1.78) but not the structure anomalies (aOR = 1.03, 95%CI 0.97-1.09). The risk of preterm delivery, low birth weight, and NICU hospitalization in their infants was increased by advanced parental age as well. Conclusion: Advanced paternal age increases the risk of congenital anomalies, especially chromosomal anomalies in their offspring, implying prenatal genetic counseling is required. What is Known: • There's a rising trend of advanced paternal age, which is associated with an increased likelihood of premature birth and low birth weight in their offspring. However, the exploration between paternal age and congenital abnormalities in offspring was limitedand contradictory. What is New: • Infants with a paternal age > 44 years were more likely to be born with congenital anomalies, especially chromosomal anomalies.

Microglia Cell Line SIM-A9 Features – New Data

SIM-A9 is a line of spontaneously immortalized mouse microglia cells obtained from newborn C57BL/6 mice’s cerebrum. The aim of this work is to characterize SIM-A9 line by the ratio of cells with the resting and activated microglia phenotype, to analyze the expression of stem/progenitor cell markers CD133 and nestin, growth factors receptors CSF-1R and EGFR, and the karyotype of this line. The light microscopy, immunocytochemistry, flow cytometry and RT/PCR were used to analyze the morphology, phenotype, and gene expression levels of pro-inflammatory cytokines, and the mFISH method was used to analyze the karyotype. It was shown for the first time that SIM-A9 cells express a high level of TSPO protein, CD68, CD11b and CD45 markers on the surface membrane of cells, which corresponds to the phenotype of activated microglia. Despite this, the cells of this line respond with additional activation to LPS stimulation, which leads to an increase in the pro-inflammatory cytokine genes IL-1β, TNFα, IL-6 expression and a high level of active oxygen and nitrogen metabolites formation. It was shown that SIM-A9 cells express stem and progenitor cells markers, CD133+ and nestin, which allows us to consider the cells of this line as early poorly differentiated progenitor cells, despite their phenotype corresponding to activated microglia. It was also found that SIM-A9 cells express receptors of two growth factors CSF-1 and EGF, CSF-1R and EGFR, which indicates the possibility of SIM-A9 cells proliferation stimulation by two alternative mechanisms under the action of the corresponding factors. SIM-A9 cells have a hypotetraploid karyotype with a large number of structural and quantitative chromosome anomalies.

Genotoxic and cytotoxic effects of thiomersal and paracetamol combination

Background: Thiomersal is used as a preservative of some vaccines or as a trace from the pathogen inactivation process in vaccine production. Prophylactic use of paracetamol upon vaccination is still common, even though paracetamol decreases immune response on some vaccines. Considering the cytotoxic and genotoxic potential of thiomersal and paracetamol and possible interaction in vivo, in vitro study was performed. Methods: The genotoxic and cytotoxic effects of thiomersal and paracetamol combination were examined on human lymphocyte cultures by using two methods: analysis of chromosomal aberrations and cytokinesis-block micronucleus cytome assay. Blood samples of three healthy donors were analyzed with the following concentrations of tested substances: thiomersal 1 µg/ml and 0.5 µg/ml, paracetamol 20 µg/ml, thiomersal 0.5 µg/ml with paracetamol 20 µg/ml and thiomersal 1.0 µg/ml with paracetamol 20 µg/ml. Results: The analysis of structural chromosomal aberrations was significantly increased in all treated cultures. In cells treated with the combination of thiomersal 1 µg/ml and 20 µg/ml of paracetamol, the number of aberrations was significantly decreased. Cytokinesis-block micronucleus cytome assay analyses showed significantly increased micronucleus frequency in lymphocytes cultivated with thiomersal 1 µg/ml compared to lymphocytes cultures exposed to thiomersal 0.5 µg/ml. Conclusions: Induction of structural chromosome aberrations and micronucleus is shown as a sign of genotoxicity for the examined concentrations of thiomersal and paracetamol. The suppressing effect of paracetamol on thiomersal genotoxicity in lymphocytes culture treated with thiomersal was shown to be indicative of further examination of paracetamol use in the prevention of genotoxicity.

Exploring X Chromosomal Aberrations in Ovarian Cells by using Fluorescence In Situ Hybridization.

Millions of people worldwide deal with issues concerning fertility. Reduced fertility, or even infertility, may be due to many different causes, including genetic disorders, of which chromosomal abnormalities are the most common. Fluorescence in situ hybridization (FISH) is a well-known and frequently used method to detect chromosomal aberrations in humans. FISH is mainly used for the analysis of chromosomal abnormalities in the spermatozoa of males with numerical or structural chromosomal aberrations. Furthermore, this technique is also frequently applied in females to detect X chromosomal aberrations that are known to cause ovarian dysgenesis. However, information on the X chromosomal content of ovarian cells from females with X chromosomal aberrations in lymphocytes and/or buccal cells is still lacking. The aim of this study is to advance basic research regarding X chromosomal aberrations in females, by presenting two methods based on FISH to identify the X chromosomal content of ovarian cells. First, a method is described to determine the X chromosomal content of isolated ovarian cells (oocytes, granulosa cells, and stromal cells) in non-grafted ovarian cortex tissue from females with X chromosomal aberrations. The second method is directed at evaluating the effect of chromosomal aberrations on folliculogenesis by determining the X chromosomal content of ovarian cells of newly formed secondary and antral follicles in ovarian tissue, from females with X chromosomal aberrations after long-term grafting into immunocompromised mice. Both methods could be helpful in future research to gain insight into the reproductive potential of females with X chromosomal aberrations.

Flavouring Group Evaluation 217 Revision 3 (FGE.217Rev3): consideration of genotoxic potential for α,β-unsaturated ketones and precursors from chemical subgroup 4.1 of FGE.19: lactones.

The Panel on Food Additives and Flavourings of the European Food Safety Authority was requested to evaluate the genotoxic potential of four flavouring substances [FL-no: 10.023, 10.030, 10.057 and 13.012] from subgroup 4.1 of FGE.19. For three of these substances [FL-no: 10.023, 10.030 and 13.012], the concern for genotoxicity has been ruled out in previous revisions of Flavouring Group Evaluation 217 (FGE.217). However, in FGE.217Rev2, a concern for genotoxicity could not be ruled out for 3a,4,5,7a-tetrahydro-3,6-dimethylbenzofuran-2(3H)-one [FL-no: 10.057]. After publication of FGE.217Rev2, industry provided additional genotoxicity studies for [FL-no: 10.057], which are evaluated in the present opinion FGE.217Rev3. The flavouring substance [FL-no: 10.057] did not induce gene mutations or numerical or structural chromosomal aberrations in vitro. Based on these data, the Panel concluded that the concern for genotoxicity is ruled out for [FL-no: 10.057]. Consequently, it can be evaluated through the Procedure.

Prenatal diagnosis and genetic analysis of two cases of Turner syndrome due to isodicentric Xp11.22

To explore the genetic characteristics of idic(X)(p11.22) in Turner syndrome (TS). Two fetuses suspected for sex chromosome abnormalities or ultrasound abnormalities were selected from Chengdu Women's and Children's Central Hospital in October 2020 and June 2020, and amniotic fluid samples were collected for G-banded chromosomal karyotyping analysis, chromosomal microarray analysis (CMA), and fluorescence in situ hybridization (FISH). The two fetuses were respectively found to have a karyotype of 45,X[47]/46,X,psu idic(X)(p11.2)[53] and 46,X,psu idic(X)(p11.2). CMA found that both had deletions in the Xp22.33p11.22 region and duplications in the p11.22q28 region. FISH showed that the centromeres in both fetuses had located on an isochromosome. The combination of karyotyping analysis, FISH, and CMA is useful for the delineation of complex structural chromosomal aberrations. High-resolution CMA can accurately identify chromosomal breakpoints, which can provide a clue for elucidating the mechanism of chromosomal breakage and rearrangement.

Characterization, Preclinical Efficacy and Toxicity Evaluations of Flavonoids Glycosides based Standardized Fenugreek Seed Extract (FEFLG)

Introduction: Fenugreek seeds, a natural food chain raw material, is known to have many flavonoid glycosides. Objective: Characterization, preclinical efficacy, and safety evaluation of flavonoid glycosidebased standardized fenugreek seed extract (FEFLG). Methods: FEFLG was characterized for a group of flavonoid glycoside marker compounds by HPLC. The CD38+ enzyme inhibition efficacy was assessed in vitro. In addition, acute oral toxicity (AOT) and subchronic, 90-day repeated-dose oral toxicity (in vivo), mutagenicity (AMES test, in vitro) and chromosome aberration test (in vitro) of FEFLG were evaluated. Results: The FEFLG was found to have 49.85% of total flavonoid glycosides content in FEFLG (25.15% of Group 1: vitexin, isovitexin and vitexin 2-o- rhamnoside and 24.70% of Group 2 (vicenin derivatives, schaftoside, iso-schaftoside, orientin and iso-orientin). FEFLG showed CD38+ enzyme inhibition in vitro (IC50= 0.96 μg/ml) equivalent to the positive control, apigenin. FEFLG did not show any toxicity at an acute oral dose of more than 2000 mg/kg (median lethal dose, LD50) with a limit dose of 5000 mg/kg. The 90-day repeated-dose oral administration of FEFLG did not induce significant toxicological changes till the maximum dose of 1000 mg/kg in male and female rats, indicating no observed adverse effect level, NOAEL ≥ 1000 mg/kg. FEFLG did not show mutagenicity (up to a concentration of 5000 μg/plate) or structural chromosomal aberrations (up to 5000 μg /ml). Conclusion: The CD38+ enzyme inhibitor efficacy in vitro, oral safety in vivo and absence of mutagenicity or genotoxicity of FEFLG indicated its potential for anti-aging applications.

Mutation Profile of Normal and Tumor Cells in a Patient with Multiple Myeloma: A Case Report

This paper is a case report of a patient with newly diagnosed multiple myeloma (MM) who underwent exome sequencing of peripheral blood lymphocytes and CD138+ tumor plasma cells prior to therapy. This patient showed some inherited genetic variants which are associated with underlying risk for MM. This patient’s genotype was reported to have some variants in the DNA repair genes, including inherited mutations in the RFDW3 and TP53 genes. They are involved in the maintenance of genome stability and accumulation rate of somatic mutations, including structural rearrangements and chromosome aberrations. A large number of structural variations&#x0D; and mutational signature ID6 in the tumor genetic material point to the disruption of DNA damage repair. The tumor cell exome analysis yielded a profile of somatic mutations, also the mutations in the genes previously associated with MM, as well as a functional significance of the detected abnormalities. Somatic mutations also included damaging mutations and highly significant mutations in the other tumor-associated genes, such as ASCC3, TET3, and CHD1, as well as in the antimicrobial peptide-coding genes CAMP and HTN3. With the exception of an extra copy of 1q arm in the tumor plasma cell genome, the patient showed no genetic risk factors associated with poor prognosis of the disease. Based on literature, inherited (ABCB1 mutations) and somatic (trisomy 3) variations detected in the patient’s genetic material can be characterized as positive prognostic factors in MM.