DNA Analysis Research Articles

Genetic Polymorphisms and Forensic Parameters of Thirteen X-Chromosome Markers in the Iraqi Kurdish Population.

X-chromosome short tandem repeat (X-STR) tools are crucial in forensic genetics and human population fields. This study presents the development and validation of a multiplex STR system consisting of thirteen X-STR loci and amelogenin specific to the human X chromosome. The system was optimized and tested for species specificity, sensitivity, stability, and DNA mixture using 9947A female and 9948 male control genomic DNA. The amplified products of nine loci were sequenced to determine the correct amplicon length. Allele frequencies, forensic parameters, mean exclusion chance (MEC), linkage disequilibrium (LD), and allelic patterns were investigated using DNA samples from 225 (159 male, 66 female) unrelated Kurdish individuals who live in Sulaymaniyah province in the Kurdistan region of Iraq. The most informative locus in the Kurdish population was GATA172D05, while the least informative locus was DXS10164. The results demonstrated that the 13 X-STR system is highly polymorphic and sensitive for forensic DNA identification. Genetic distance-based clustering, metric multidimensional scaling (MDS), and correlation matrix were analyzed for 19 ethnic groups and populations. The phylogenetic tree showed that populations clustered according to their ethnogeographic relationships. The findings revealed genetic links between the Iraqi Kurds, Caucasians, Iraqi Arabs, United States (U.S.) ethnic groups, and Chinese populations.

DNA-targeting short Argonautes complex with effector proteins for collateral nuclease activity and bacterial population immunity.

Two prokaryotic defence systems, prokaryotic Argonautes (pAgos) and CRISPR-Cas, detect and cleave invader nucleic acids using complementary guides and the nuclease activities of pAgo or Cas proteins. However, not all pAgos are active nucleases. A large clade of short pAgos bind nucleic acid guides but lack nuclease activity, suggesting a different mechanism of action. Here we investigate short pAgos associated with a putative effector nuclease, NbaAgo from Novosphingopyxis baekryungensis and CmeAgo from Cupriavidus metallidurans. We show that these pAgos form a heterodimeric complex with co-encoded effector nucleases (short prokaryotic Argonaute, DNase and RNase associated (SPARDA)). RNA-guided target DNA recognition unleashes the nuclease activity of SPARDA leading to indiscriminate collateral cleavage of DNA and RNA. Activation of SPARDA by plasmids or phages results in degradation of cellular DNA and cell death or dormancy, conferring target-specific population protection and expanding the range of known prokaryotic immune systems.

Forensic Genealogy in Solving Age- Old Cases: Review

Forensic genealogy, a revolutionary technique at the intersection of criminal investigations, genetics, and genealogy, has gained recognition for solving mysteries and finding missing persons. This article provides an overview of its approach, highlighting its pivotal role in long-standing cases and the transformative impact of DNA analysis and genealogical research. It also includes case studies that demonstrate the effectiveness of forensic genealogy techniques.

The seeds of invasion: a comparison of endophytic seed bacteria of Acacia longifolia between its native and invasive ranges

Acacia longifolia is an aggressive invader in Mediterranean-type ecosystems severely impacting biodiversity and ecosystem functions. The species’ invasiveness has been linked to its ability to thrive in nutrient poor soils, high seed production, and quick establishment after fire. In this study, we identify and compare the bacterial endophytes of A. longifolia seeds collected from populations in the species’ native (Australia) and invasive (Portugal) ranges. For this, we characterised the morphology (length, width, and weight) of seeds from two sites in each range and isolated and cultivated bacteria from seeds. DNA fingerprinting and cluster analyses revealed slightly higher, and distinct, bacterial diversity associated with seeds collected from native range populations in comparison to those collected from invasive populations. Sequencing of the 16S rDNA gene identified 119 bacterial isolates from 15 genera, with Curtobacterium strains being common in both ranges. Several differences in bacterial genera were found among ranges and sites: Dermacoccus, Frigoribacterium, Kocuria, Pantoea and Phyllobacterium taxa were each unique to seeds from the native populations, while Brevundimonas, Microbacterium, Rhizobium and Sphingomonas taxa were only found in the invasive seeds. The genus Paraburkholderia occurred in all invasive-range seeds but was not isolated from the native-range. Bacillus and Paenibacillus co-occurred in seeds collected from all invaded sites, but the simultaneous presence of both taxa was not found in native-range seeds. We propose that the bacterial endophytes present in invasive-range seeds may be important players for the invasiveness of A. longifolia, due to their role as plant growth promoters, providing extra capabilities helping acacia expansion.

Comparison of DNA quantity and quality from fecal samples of mammals transported in ethanol or lysis buffer

Using fecal microbial community profiles through sequencing approaches helps to unravel the intimate interplay between health, wellness, and diet in wild animals with their environment. Ensuring the proper preservation of fecal samples before processing is crucial to ensure reliable results. In this study, we evaluated the efficiency of two different preservation methods, considering the following criteria: DNA yield, quality and integrity, and microbial community structure based on Oxford Nanopore amplicon sequencing of the V3-V4 region of bacterial 16S rRNA and protozoa 18S rRNA genes. Eighteen matched pairs of mammalian fecal samples were collected and transported in 99.8% ethanol and lysis buffer; processing occurred between 55 and 461 days post-collection. Wilcoxon signed-rank tests were used to analyze quantitative measurements for paired samples. The A260/280 ratio, a measure of nucleic acid purity, was assessed descriptively for each media, and the Bartlett test evaluated dispersion of this ratio. A Fisher test was performed to compare the number of positive reactions for DNA extraction or PCR amplification of the 16S and 18S rRNA genes between both media. The concentration of total DNA and amplicons, as well as the number of reads obtained in sequencing, was significantly higher in the samples preserved with lysis buffer compared to ethanol, with magnitudes up to three times higher. Electrophoretic analysis of total DNA and amplicons further confirmed superior DNA integrity in lysis buffer preserved samples. The A260/280 values obtained using the lysis buffer were of optimal purity (mean: 1.92) and with little dispersion (SD: 0.27); on the other hand, the ethanol samples also presented an excellent average quality (mean: 1.94), but they were dispersed (SD: 1.10). For molecular studies using mammalian feces, the lysis buffer reagent proved to be a reliable solution for their collection, conservation, and storage.

Anemia among Medical Students from Jakarta: Indonesia-Iron Deficiency or Carrier Thalassemia?

Anemia, a global health concern, affects one-fourth of the global population, particularly women. In Indonesia, its prevalence is 23.7%, with 32.0% among 15-24 year-olds. Factors include poor nutrition, infectious diseases, chronic diseases, inherited disorders, and inadequate healthcare access. This study aimed to investigate anemia prevalence and its etiology among medical students from Jakarta. This study was a descriptive research with a cross-sectional approach. Undergraduate students aged 18-23 years old were selected and consented to participate by a consecutive nonrandom sampling methods. Laboratory blood data were evaluated (including Hb, MCV, MCH, HbA2, and ferritin levels) and DNA was isolated to confirm the type of thalassemia carrier. In total, 140 medical students, mainly female, were recruited. Anemia was found in 13.6% (11.4% had low MCV and/or MCH), and 16.5% had low MCV and/or MCH without anemia. Hb electrophoresis revealed high HbA2 values, suggesting the HbE variant (2.1%), and β-thalassemia carrier (0.7%). DNA analysis confirmed the cd26 mutation and heterozygous IVS1nt5. Among those without anemia, 5% had α-deletion, while in the group with anemia, 1.4% had α-deletion (with coexistent IDA), 3.6% had α-deletion, and 0.7% had β-mutation. DNA analysis can identify specific mutations associated with alpha-thalassemia, distinguishing between iron deficiency anemia and the alpha-thalassemia trait. Thalassemia screening should involve low MCV and/or MCH values as the first step (stage 1), followed by Hb analysis (stage 2) and DNA analysis (stage 3). In common areas, a combination of Hb and DNA testing is best. However, healthcare professionals must diagnose and treat thalassemia, as proper management relies on accurately identifying the underlying condition.

Application of X-STRs for Forensic Identification in Mixed DNA Profile: A Case Report

Autosomal polymorphisms (STRs) or Y-Chromosome polymorphisms (Y-STRs) are usually used for the study and deconvolution of mixed DNA profiles in forensic genetics, accompanying data interpretation with biostatistical evaluations (e.g., RMP, RMNE, LR). Sometimes, however, some mixed DNA profiles are so complex that autosomal and Y markers are not sufficient for correct discrimination and identification. In this work is reported a robbery case in which the analysis of the polymorphic markers of the X Chromosome (X-STRs) was applied to the mixed profiles obtained from the traces. This falls outside the classic use of the X-STRs. Indeed, the aim of the authors is to encourage the usage of X-STRs not only in parental relationships, but also in pure forensic cases for interpreting complex mixed DNA profiles, since their application in case resolution could be more decisive than autosomal STRs and Y-STRs.

Investigating Whether the Improvement of the AMP-Activated Protein Kinase (AmpK) is Neuroprotective in Huntington’s Disease Neurons

Huntington’s disease (HD) is a hereditary condition which causes disordered movement, behavioral changes, and dementia. As with many neurological diseases, there is no effective treatment and HD is universally fatal. Metformin has been found improve cognitive score in diabetic patients with HD and has also improved motor function and decreased mutant HTT aggregates in mice brains. Like metformin, BC1618 is an inhibitor of Fbxo48 and its potency in stimulating Ampk-dependent signaling greatly exceeds metformin. GA100 also works in a similar manner as BC1618. Thus, Q7 and Q111 cell lines were treated with 5 and 10 μM concentrations of BC1618 or GA1000 and the samples were collected to perform protein, RNA, and DNA analysis. The most significant results from this experiment came from the qPCR results which confirmed the initial hypothesis and supported the idea that both BC1618 and GA100 effectively inhibit Fbxo48 and prolong Ampk activation. We further hypothesize that this will promote mitochondrial biogenesis which can have neuroprotective effects on HD.

Y-STR analysis of highly degraded DNA from skeletal remains over 70years old.

The goal of the following study is to clarify whether the skeletal remains over 70years old from missing persons and their alleged relatives shared identical Y-STR loci. Nowadays, advances in ancient DNA extraction techniques and approaches of using multiple different Y-STRs have significantly increased the possibility of obtaining DNA profiles from highly degraded skeletal remains. Given the ages and conditions of the skeletal remains, ancient DNA extraction methods can be used to maximize the probability of DNA recovery. Considering that information about distant relatives is more relevant for long-term missing persons and alleged family members are male, Y-STR loci analysis is considered the most appropriate and informative approach for determining paternal lineage relationship. In this study, Y-STR genotypes obtained from these alleged relatives were identical to each other and to the alleles of missing persons' consensus profiles at more than 22 loci examined, whilst not being found in Y-STR population database from Y-Chromosome STR Haplotype Reference Database. Therefore, Missing Person No.7 and Missing Person No.18 have a patrilineal relationship with reference samples from Family1 and Family2, respectively. In addition, the fact that Y-STR haplotypes obtained from skeletal remains of missing persons and reference samples are not found in the Han Chinese people from East Asian demonstrates its rarity and further supports a paternal lineage relationship amongst them.

Genomic spectrum of actionable alterations in serial cell free DNA (cfDNA) analysis of patients with metastatic breast cancer

We aimed to study the incidence and genomic spectrum of actionable alterations (AA) detected in serial cfDNA collections from patients with metastatic breast cancer (MBC). Patients with MBC who underwent plasma-based cfDNA testing (Guardant360®) between 2015 and 2021 at an academic institution were included. For patients with serial draws, new pathogenic alterations in each draw were classified as actionable alterations (AA) if they met ESCAT I or II criteria of the ESMO Scale for Clinical Actionability of Molecular Targets (ESCAT). A total of 344 patients with hormone receptor-positive (HR+)/HER2-negative (HER2-) MBC, 95 patients with triple-negative (TN) MBC and 42 patients with HER2-positive (HER2 + ) MBC had a baseline (BL) cfDNA draw. Of these, 139 HR+/HER2-, 33 TN and 13 HER2+ patients underwent subsequent cfDNA draws. In the HR+/HER2- cohort, the proportion of patients with new AA decreased from 63% at BL to 27–33% in the 2nd-4th draws (p < 0.0001). While some of the new AA in subsequent draws from patients with HR+/HER2- MBC were new actionable variants in the same genes that were known to be altered in previous draws, 10-24% of patients had new AA in previously unaltered genes. The incidence of new AA also decreased with subsequent draws in the TN and HER2+ cohorts (TN: 25% to 0–9%, HER2 + : 38% to 14–15%). While the incidence of new AA in serial cfDNA decreased with subsequent draws across all MBC subtypes, new alterations with a potential impact on treatment selection continued to emerge, particularly for patients with HR+/HER2- MBC.

A first report of Pseudosuccinea columella (Say, 1817), an alien intermediate host for liver fluke, in Malawi

Starting in October 2021, quarterly malacological surveys have been undertaken in Malawi, with the sampling of 12 specified freshwater habitats throughout a calendar year. Each survey monitors the presence of aquatic intermediate snail hosts of medical and veterinary importance. In March 2023, the alien lymnaeid species Pseudosuccinea columella was encountered for the first time in the surveys, in Nsanje District. This species identity was later confirmed upon DNA analysis of mitochondrial ribosomal 16S sequences. In July 2023, P. columella was also noted at single sites within Mangochi and Chikwawa Districts, and again in Nsanje District, with an additional location observed. Of particular importance, our sampled location in Mangochi District was directly connected to Lake Malawi, which expands the species list of invasive molluscs in this lake. While P. columella is a well-known intermediate snail host for human and animal fascioliasis, screening collected snails for trematode cercariae, alongside molecular xenomonitoring, did not yield equivocal evidence of active fluke infection. However, the newly recognized presence of this alien intermediate snail host within Lake Malawi, and along the Shire River Valley, flags a new concern in altered local transmission potential for human and animal fascioliasis.Graphical

Exploring the role of the human microbiome in forensic identification: opportunities and challenges.

Forensic microbiology is rapidly emerging as a novel tool for human identification. The human microbiome, comprising diverse microbial communities including fungi, bacteria, protozoa, and viruses, is unique to each individual, offering a new dimension to forensic investigations. While traditional identification methods primarily rely on DNA profiling and fingerprint analysis, they face limitations when complete DNA or fingerprints profiles are unattainable or degraded. In this context, the microbial signatures of the human skin microbiome present a promising alternative due to their resilience to environmental stresses and individual-specific composition. This review explores the potential of microbiome analysis in forensic human identification, evaluating its applications, advantages, limitations, and future prospects. The uniqueness of an individual's microbial community, particularly the skin microbiota, can provide distinctive biological markers for identification purposes, while technological advancements like 16S rRNA sequencing and metagenomic shotgun sequencing are enhancing the specificity of microbial identification, enabling detailed analysis of these complex ecological communities. Despite these promising findings, current research has not yet achieved a level of identification probability that could establish microbial analysis as a stand-alone evidence tool. Therefore, it is presently considered ancillary to traditional methods, contributing to a more comprehensive biological profile of individuals.

A taxonomic revision of the Palaearctic species of the ant genus Tapinoma Mayr 1861 (Hymenoptera: Formicidae)

A taxonomic revision of the Palaearctic species of the ant genus Tapinoma Emery 1861 is provided. Due to the extreme rarity of discernable characters, the female castes of 64% of the species are not separable by primary visual perception of an expert and are thus truly cryptic. The main working rationale of the revision is Numeric Morphology-Based Alpha-Taxonomy (NUMOBAT) with formation of species hypotheses largely based on exploratory data analyses and checking these hypotheses by discriminant analysis. NUMOBAT data of all considered species comprise 648 worker samples with 2014 individuals and 30,000 primary data. Species are depicted by z-stack imaging in three standard visual positions. Numeric data on 15 phenotypical characters are presented in comparative tables and supplementary verbal descriptions are given. Analysis of nuclear DNA was based on evaluation of 15 microsatellite loci in 274 nest samples. The NUMOBAT-based classification of eight species of the Tapinoma nigerrimum and T. simrothi group which are inseparable by subjective visual inspection was confirmed by nuclear DNA data in 98.3% of 169 samples where both morphological and nuDNA data were available. The three cases of disagreement between NUMOBAT and nuDNA classification were discussed. Excluding Nomina Nuda and unavailable names, the revision recognized 34 available names which divide into 21 valid species, 11 junior synonyms and two names placed under Incertae Sedis. Four species are described as new: Tapinoma hispanicum n. sp., T. insularis n. sp., T. dabashanica n. sp. and T. sichuense n. sp. The following synonymies were stated: T. collina Foerster 1850, T. erraticum platyops Emery 1925 and T. erraticum bononiensis Emery 1925 as synonyms of T. erraticum (Latreille 1798); T. tauridis Emery 1925 and T. breve Emery 1925 as synonyms of T. glabrella (Nylander 1849) that is erected to species level; T. minor Bernard 1945 as synonym of T. simrothi Krausse-Heldrungen 1911; T. simrothi azerbaidzhanica Karavajev 1932 as synonym of Tapinoma karavaievi Emery 1925 that is erected here to species level; T. erraticum ambiguum Emery 1925 as synonym of T. madeirense Forel 1895 and T. emeryanum Kuznetzov-Ugamsky 1927 as synonym of T. sinense Emery 1925. Despite an insufficient separation of the female castes of Tapinoma madeirense and T. subboreale Seifert 2011 by NUMOBAT characters and detection of a hybrid zone in southern France, the heterospecific status of the two taxa was maintained based on sufficient differences in male genitalia and of nuclear DNA over a larger geographic scale. Formica caerulescens Losana 1834 is removed from genus Tapinoma and recognized as member of the subfamily Formicinae.

Silver nanoparticle-induced in vitro flowering in Dendrocalamus strictus (Roxb.) nees and genetic fidelity assessment of regenerants using molecular markers.

Dendrocalamus strictus (Roxb.) Nees, generally referred to as 'Male bamboo,' is a globally prevalent and highly significant species of bamboo. It is a versatile species and possesses notable industrial significance. However, despite its numerous applications, the production of this plant is insufficient to fulfill the worldwide demand. The challenges that impede the dissemination of D. strictus encompass the unpredictable blooming pattern (30-70 years), low seed production, and limited seed viability. Therefore, tissue culture presents a reliable and effective option for the mass production of standardized planting material. This study investigated the effects of silver nanoparticles (AgNPs) at a concentration of 6.0 mg L- 1 in the Murashige and Skoog (MS) nutrient medium fortified with pre-optimized plant growth regulators (3.0 mg L- 1 6-benzylaminopurine + 0.5 mg L- 1 α-naphthalene acetic acid) on the induction of flowering in a controlled environment in D. strictus. The use of AgNPs in the media induced a maximum of 14 inflorescences per culture vessel, 9 flowers per inflorescence, and improved the performance of the micropropagated plantlets during acclimatization in the greenhouse and field. The ISSR and SCoT amplified polymorphic DNA analysis of the regenerants resulted in the formation of 49 bands (300 to 2000bp size) and 36 scorable bands (350 to 2000bp) respectively. All the PCR amplicons produced by SCoT and ISSR were monomorphic confirming the genetic uniformity of the tissue cultured plants of D. strictus with the mother plant. It can be inferred that the incorporation of AgNPs during the shoot proliferation phase has the potential to stimulate in vitro flowering in D. strictus. This finding could provide valuable insights into innovative strategies for enhancing crop productivity and genetic manipulation for accelerated breeding and agricultural advancement.

CGAS suppresses hepatocellular carcinoma independent of its cGAMP synthase activity.

Cyclic GMP-AMP synthase (cGAS) is a key innate immune sensor that recognizes cytosolic DNA to induce immune responses against invading pathogens. The role of cGAS is conventionally recognized as a nucleotidyltransferase to catalyze the synthesis of cGAMP upon recognition of cytosolic DNA, which leads to the activation of STING and production of type I/III interferon to fight against the pathogen. However, given that hepatocytes are lack of functional STING expression, it is intriguing to define the role of cGAS in hepatocellular carcinoma (HCC), the liver parenchymal cells derived malignancy. In this study, we revealed that cGAS was significantly downregulated in clinical HCC tissues, and its dysregulation contributed to the progression of HCC. We further identified cGAS as an immune tyrosine inhibitory motif (ITIM) containing protein, and demonstrated that cGAS inhibited the progression of HCC and increased the response of HCC to sorafenib treatment by suppressing PI3K/AKT/mTORC1 pathway in cellular and animal models. Mechanistically, cGAS recruits SH2-containing tyrosine phosphatase 1 (SHP1) via ITIM, and dephosphorylates p85 in phosphatidylinositol 3-kinase (PI3K), which leads to the suppression of AKT-mTORC1 pathway. Thus, cGAS is identified as a novel tumor suppressor in HCC via its function independent of its conventional role as cGAMP synthase, which indicates a novel therapeutic strategy for advanced HCC by modulating cGAS signaling.

Comparison of the Efficacy of Two Routes of Administration of Human Amniotic Epithelial Cells in Cell Therapy of Acute Hepatic Insufficiency.

The route of administration of implanted cells may affect the outcome of cell therapy by directing cell migration to the damaged site. However, the question of the relationship between the route of administration, the efficacy of colonisation of a given organ, and the efficacy of cell therapy has not been resolved. The aim of the study was to localise transplanted intravenously and intraperitoneally human amniotic epithelial cells (hAECs) in the tissues of mice, both healthy and injured, in an animal experimental model of acute liver failure (ALF). Mice intoxicated with D-Galactosamine (D-GalN) at a dose of 150 mg/100 g body weight received D-GalN alone or with a single dose of hAECs administered by different routes. Subsequently, at 6, 24, and 72 h after D-GaIN administration and at 3, 21, and 69 h after hAEC administration, lungs, spleen, liver, and blood were collected from recipient mice. The degree of liver damage and regeneration was assessed based on biochemical blood parameters, histopathological evaluation (H&E staining), and immunodetection of proliferating (Ki67+) and apoptotic (Casp+) cells. The biodistribution of the administered cells was based on immunohistochemistry and the identification of human DNA. It has been shown that after intravenous administration, in both healthy and intoxicated mice, most of the transplanted hAECs were found in the lungs, while after intraperitoneal administration, they were found in the liver. We concluded that a large number of hAECs implanted in the lungs following intravenous administration can exert a therapeutic effect on the damaged liver, while the regenerative effect of intraperitoneally injected hAECs on the liver was very limited due to the relatively lower efficiency of cell engraftment.

The Value of Forensic DNA Database

This paper explores the importance and impact of DNA databases in forensic investigations. It responds to a recent publication by Kostiuchenko, Vynohradova, Sereda, Svoboda, and Polunina (2024), highlighting challenges in setting up and using DNA databases. These challenges include logistical, staffing, legal, security, growth, public interest, and international cooperation issues. By examining South Africa's successful NFDD implementation, the authors stress the need for thorough planning and implementing a comprehensive value chain. Drawing insights from South Africa's successful NFDD implementation, the authors emphasise the importance of meticulous planning and implementing a value chain. This value chain should cover DNA evidence collection, follow-up investigations, and presenting evidence in court.

Distribution of eukaryotic environmental DNA in global subseafloor sediments

The analysis of eukaryotic environmental DNA (eDNA) in sediment has the potential for understanding past ecosystems, even for taxa lacking skeletons or preserved only as a part of necromass. Despite the paleoenvironmental and ecological importance of eukaryotic eDNA in marine sediment, the duration of remaining eDNA and the species of eDNA present in marine sediment has not been well investigated. Here, we analyzed eDNA extracted from 299 sediment samples down to 678 m below the seafloor at 40 geologically distinct sites. The results showed that eukaryotic eDNA was amplified from more than 80% of the sediments with a depositional age of less than 100,000 years. The eDNA was well conserved in anoxic sediments than in oxic sediments, with PCR success rates of 48% and 18%, respectively. The eukaryotic communities include non-benthic organisms such as marine plankton, including diatoms, dinoflagellates, and coccolithophores. A freshwater diatom genus Aulacoseira was detected in the Baltic Sea sediments from the last glacial lacustrine environment. These results provide new insights into the global-scale distribution of the past eukaryotic eDNA preserved in marine sediment.

Evaluation of the effect of COMT gene polymorphism in rs4680 on the occurrence of concomitant head, neck and back pain in patients with temporomandibular joint disorders

Background: Temporomandibular disorders (TMD) include a number of clinical conditions involving the masticatory muscles, temporomandibular joint (TMJ), and adjacent structures. The main feature of TMD is pain. According to past studies, TMD can have genetic causes. One of the genes that seems to be important in this regard is COMT. This study aims to compare the effect of COMT gene polymorphism rs4680 on the occurrence of simultaneous head, neck and back pain in patients with TMD.&#x0D; Methods: This study was conducted as a case-control study on patients aged between 18 and 65 with TMD disorder. After meeting the conditions for entering the study, a written informed consent was obtained from the people participating in this project. Then a blood sample of 5cc was prepared from each person and poured into tubes containing EDTA anticoagulant and used for DNA extraction and identification. The COMT gene was sent to the genetics laboratory and was compared in terms of the polymorphism of the COMT gene using the PCRARMS technique.&#x0D; Results: The sample size was 100 people aged 18 to 65 years. Variables with normal distribution were analyzed with Chi-Square test and variables with non-normal distribution with Mann-Whitney test. After genetic analysis in terms of rs4680 gene polymorphism, 18 people (18%) were identified with AA, 80 people (80%) with GA and 2 people (2%) with GG. "temporal headache in the last 30 days", "temporal headache change with chewing hard foods", "temporal headache change with opening the mouth or moving the jaw forward or to the sides", " Temporal headache changes with jaw habits such as teeth touching, pressing or grinding teeth together, chewing gum ", " Temporal headache changes with jaw activities such as talking, kissing or yawning ", " having Back pain during the last 4 weeks", having pain in the arm, leg, joints (neck) during the last 4 weeks", “headache during 4 last weeks” and "time of onset of headache in the temporal region" did not have significant relationships with COMT gene polymorphism rs4680 in patients with TMD.(repectively, p value=0.873، p value=0.658، p value=0.518 p value=0.685، p value=0.884،p value=0.654، p value=0.723، p value=0.831، p value=0.692 ).&#x0D; Conclusion: According to previous studies, there is a significant relationship between rs4680 polymorphism in the COMT gene and the occurrence of temporomandibular disorders (TMD), although in our study, no significant relationship was found between headache and neck and back pain with this polymorphism.

Effect of Silver-Graphene Oxide-Cobalt Oxide Nanocomposite on Cytotoxic Levels in MRC-5 and HepG2 Cell Lines and Molecular Docking Studies

The cytotoxic properties of cobalt oxide (Co3O4) nanoparticles (NPs), in addition to graphene oxide (GO)-Co3O4 and silver (Ag)-GO-Co3O4 nanocomposites (NCs), were evaluated against both human healthy lung fibroblast (MRC-5) and hepatocellular carcinoma (HepG2) cell lines utilizing the XTT assay. The investigation revealed that synthesized Co3O4 NPs and NCs (GO-Co3O4 and Ag-GO-Co3O4) elicited significant cytotoxic responses in MRC-5 and HepG2 cell lines in a concentration-dependent manner. Through molecular docking analyses, it was observed that all fabricated nanomaterials exhibited DNA recognition via minor groove binding, with molecular affinities ranging from − 4.82 to -11.66 kcal/mol. Furthermore, the docking outcomes illustrated that the angular conformations of GO-Co3O4 and Ag-GO-Co3O4 conferred ‘shape-selective’ characteristics as DNA minor groove binders, leading to heightened cytotoxicity, particularly in the HepG2 cell line compared to the normal MRC-5 cell line.