Deoxyribonucleic Acid Analysis Research Articles

Research on Correlation between DNA Typing and Trace Characteristics of Blood after Thermal Exposure

The identification of biological evidence is particularly important for criminal detection, and the deoxyribonucleic acid (DNA) analysis plays a significant role in reconstructing events. However, bloodstains after thermal exposure in fires are quite unique compared to those in other scenes. Previous results regarding DNA recovery in bloodstains after heating are inconsistent with each other, which limits guidance for forensic science. In order to confirm the influence of heat on DNA recovery, the important physical evidence, bloodstain, was selected for its common occurrence, and a standard heat source, the Cone Calorimeter, was used to simulate high temperatures in fire scenes. A series of bloodstains were prepared under different heating conditions, and the results of short tandem repeat (STR) typing were systematically correlated with the trace characteristics of bloodstains. The findings indicate that heating bloodstains below 150oC for less than 10mins has minimal effect on DNA testing. After heating bloodstains at 150oC for 20mins or at 180oC for 5mins, partial DNA profiles were obtained, accompanied by blackening and cracking of the bloodstains. After exposure to 180oC for 20mins or 200oC for 10mins, no DNA profiles were obtained with bloodstains exhibiting metallic lusters and black bulges. Furthermore, from the perspective of chemical bond energy, the C-N, C-O, C-C, and P-O bonds in DNA molecules are prone to breaking during heating. The C-N bond serves as the primary connection between the four bases and the strand, while the C-O, C-C, and P-O bonds are significant connections within the DNA strand. It is thus hypothesized that the breakage of any bond aforementioned during heating results in the failure of DNA typing. Understanding the correlation between trace characteristics of bloodstains and DNA typing results after thermal exposure is crucial for comprehending DNA recovery from physical evidence collected from fire scenes.

Hypomethylation of the RUNX2 Gene Is a New Potential Biomarker of Primary Osteoporosis in Men and Women.

The search for the molecular markers of osteoporosis (OP), based on the analysis of differential deoxyribonucleic acid (DNA) methylation in bone cells and peripheral blood cells, is promising for developments in the field of the early diagnosis and targeted therapy of the disease. The Runt-related transcription factor 2 (RUNX2) gene is one of the key genes of bone metabolism, which is of interest in the search for epigenetic signatures and aberrations associated with the risk of developing OP. Based on pyrosequencing, the analysis of the RUNX2 methylation profile from a pool of peripheral blood cells in men and women over 50 years of age of Russian ethnicity from the Volga-Ural region of Russia was carried out. The level of DNA methylation in three CpG sites of the RUNX2 gene was assessed and statistically significant hypomethylation was revealed in all three studied CpG sites in men (U = 746.5, p = 0.004; U = 784, p = 0.01; U = 788.5, p = 0.01, respectively) and in one CpG site in women (U = 537, p = 0.03) with primary OP compared with control. In the general sample, associations were preserved for the first CpG site (U = 2561, p = 0.0001766). The results were obtained for the first time and indicate the existence of potentially new epigenetic signatures of RUNX2 in individuals with OP.

Unsteady solute dispersion of electro-osmotic flow of micropolar fluid in a rectangular microchannel

This study scrutinizes the two-dimensional concentration distribution for a solute cloud containing a micropolar fluid in a rectangular microchannel under the influence of an applied electric field. The concentration distribution is obtained up to second order approximation using Mei's homogenization method. Analytical formulas are derived for dispersion coefficient, mean and two-dimensional concentration distributions. This study also includes the analytical expressions for electric potential, velocity, and microrotation profiles. This study discusses the impact of coupling number, couple stress parameter, electric double layer thickness, and Péclet number on solute concentration distribution. The results of fluid velocity and dispersion coefficient are validated with available works in the literature. The non-Newtonian parameter and electric double layer thickness are shown to have a significant impact on dispersion. Our study reveals that concentration distribution rises but spreading of solute reduces when the coupling number increases. This is also true when the Debye length decreases. It is also obtained that the solute spreads more in the Newtonian fluid case compared to the micropolar fluid case. Finally, coupling number and electric double layer thickness show a symmetric pattern to the indicator function for the transverse concentration variation rate. The findings of this work have broad implications in deoxyribonucleic acid analysis, chemical mixing, and separation.

Culture time to optimize embryo cell-free DNA analysis for frozen-thawed blastocysts undergoing noninvasive preimplantation genetic testing for aneuploidy

To investigate the ideal time in culture to optimize embryo cell-free deoxyribonucleic acid (cfDNA) analysis in frozen-thawed blastocysts undergoing noninvasive preimplantation genetic testing for aneuploidy (PGT-A). Cell-free DNA is released into the spent blastocyst media (spent media) by the embryo. However, the optimal timing to determine maximal cfDNA in the case of frozen-thawed blastocysts undergoing noninvasive PGT-A remains to be elucidated. In this prospective observational study, 135 spent media and corresponding whole blastocysts were collected from January 2021 through March2022. Private fertility clinics. Day-5 frozen-thawed blastocysts were cultured for 8 hours (Day-5 Short) or 24 hours (Day-5 Long), whereas day-6 frozen-thawed blastocysts were cultured for 8 hours (Day-6 Short). The spent media and whole blastocysts were then collected for further analysis. Spent media and whole blastocysts were amplified using whole genome amplification and sequenced using next-generation sequencing. Informativity and concordance rates between cfDNA in spent media and whole blastocyst DNA were compared according to the different times in culture. When comparing time in culture, informativity rates for spent media were significantly higher for Day-5 Long and Day-6 Short (>91%) compared with the Day-5 Short group (<60%). A similar trend was observed for cases with and without a previous PGT-A. Regarding blastocyst expansion grade, informativity rates were lower on Day-5 Short compared with Day-5 Long and Day-6 Short, regardless of expansion degree. This decrease was significant for Gardner-grade expansion grades 3, 4, and 5-6. In addition, for a similar time in culture, the grade of expansion did not have an impact on the informativity rates. For concordance rates, no significant differences were observed among the 3 groups. In all cases, concordance rates were 90.5% for Day-5 Short, 93.6% for Day-5 Long, and 92.3% for Day-6 Short. No impact of the expansion grade was observed on concordance rates. Noninvasive PGT-A in frozen-thawed blastocysts yields very high concordance rates with whole blastocysts, possibly limiting the need for invasive PGT-A and making it available for a wider range of patients.

CDK4/6 Inhibitor Efficacy in ESR1-Mutant Metastatic Breast Cancer

BackgroundIn estrogen receptor–positive metastatic breast cancer, ESR1 mutations (ESR1m) are a common mechanism of acquired resistance to aromatase inhibitors (ArIh). However, the impact ESR1 alterations have on CDK4/6 inhibitor (CDK4/6i) sensitivity has not been established. Analyses of CDK4/6i trials suggest that the endocrine therapy partner and specific ESR1 allele may affect susceptibility. We analyzed a real-world data set to investigate CDK4/6i efficacy in ESR1m metastatic breast cancer and associated clinical factors.MethodsESR1m were identified by analysis of circulating-tumor deoxyribonucleic acid. The GuardantINFORM database contains genomic information from tumors linked with claims data. Patients who started a CDK4/6i within 30 days of sequencing were categorized as having ESR1m or non-ESR1–mutant (non-ESR1m) breast cancer. Data were analyzed to determine the real-world time-to-next-treatment, defined as the start of a breast cancer treatment to initiation of the subsequent treatment.ResultsOne hundred forty-five patients with ESR1m and 612 with non-ESR1m metastatic breast cancer were analyzed. ESR1m and non-ESR1m tumors had similar real-world time-to-next-treatment on CDK4/6i regimens (hazard ratio, 1.02; 95% confidence interval, 0.82 to 1.23). Duration on therapy in the first-line and second-line plus treatment settings were comparable regardless of ESR1 status. We stratified treatment duration by concurrent endocrine therapy, and patients with ESR1m had worse outcomes on ArIh but comparable real-world time-to-next-treatment on fulvestrant.ConclusionsThese data suggest ESR1 variants are not associated with pan-CDK4/6i resistance and are consistent with the hypothesis that CDK4/6 blockade combined with a selective estrogen receptor degrader is potentially an effective option for ESR1m metastatic breast cancer.

Why Down-managing Backlog Forensic DNA Case Entries Matters

Forensic laboratories face a backlog of case files, affecting service delivery, causing delays. Forensic laboratories face a backlog of case ϐiles, affecting service delivery and causing delays. The backlog points to underfunding, poor planning, and inadequate support, hindering deoxyribonucleic acid (DNA) analysis. Resolving casework backlogs may initially seem like a straightforward and attainable measure to improve the arrest of offenders and promote justice. Longer turnaround times impede investigative leads, emphasising the need for efficient strategies and a comprehensive approach to address and prevent backlogs in forensic laboratories. No study has been published on the forensic DNA backlogs in South Africa. The article explicitly addresses one aspect of a Doctor of Philosophy study and aims to ascertain the impact of backlogs of forensic DNA case entries. The study article’s research questions included the following: “What cases are considered as backlog?”; “What is the current backlog in forensic DNA case entries in South Africa?” and “What are the main reasons for the backlog of cases involving forensic DNA?” The prompt processing of DNA evidence is vital not only for safeguarding individuals falsely accused of crimes based on circumstantial evidence but also for adding prosecutors and providing justice for crime victims.

Label-free identification of DNA single-base mutation for clinical diagnostics based on plasmonic SiNW@AgNP arrays

Surface-enhanced Raman spectroscopy (SERS) has exhibited great potential in deoxyribonucleic acid (DNA) analysis. Rapid and label-free identification of single-base mutation in DNA is still limited by the sensitivity and reproducibility of SERS substrates. In this work, we develope plasmonic SiNW arrays decorated with Ag nanoparticles (SiNW@AgNP arrays) via hexadecyltrimethylammonium bromide (CTAB)-assisted galvanic reaction to address these challenges, which show excellent sensitivity and reproducibility. Under optimized experimental conditions, plasmonic SiNW@AgNP arrays enable detection of DNA bases of adenine (A), guanine (G), cytosine (C), and thymine (T) in solution at concentration down to pM-μM range, and multiple detection ability with mixture of DNA bases. More significantly, our study demonstrates the SiNW@AgNP arrays may serve as excellent SERS substrates in discriminating DNA single-base mutation between sickle cell anemia gene and normal hemoglobin gene in serum samples, which can be exploited in a wide range of plasmonic enhanced sensing applications for a fast, real-time, label-free and cost-effective clinical diagnosis.

Significance and challenges of DNA profiling in medicolegal setup of Pakistan

Deoxyribonucleic acid (DNA) serves as a unique genetic blueprint, inherited from our parents, making it an ideal identifier of individuals. The application of DNA analysis has revolutionized the field, leading to accurate and reliable results that have helped solve complex cases and bring justice to victims. This article focuses on the DNA-based reports in criminal justice system, as well as the accompanying challenges, emphasizing the necessity for particular DNA-based laws. The DNA discovery opened up new paths and the improvement of DNA identification and its incorporation into legal system has offered significant assistance in settlement of criminal and civil matters. Primarily, DNA proofing is used in a paternity dispute in Pakistan. Overall, the integration of DNA analysis into forensic science has significantly enhanced the process of investigating and resolving criminal cases. However, the necessity for laws and standards to allow its use for forensic reasons in Pakistan has long been recognized.

A comparative methodological approach to studying the diet of a recovering marine predator, the grey seal (Halichoerus grypus)

Anthropogenic influences caused depletion and subsequent recovery of marine predators, but ecological consequences of altered predator abundance are not well understood. Although many methods are used to study predator diets, methodological biases and logistical challenges preclude robust sampling schemes. We aimed to compare two non-invasive methods: metabarcoding scat-derived deoxyribonucleic acid and hard parts analysis of scat for the Northwest Atlantic grey seal ( Halichoerus grypus (Fabricius, 1791)), a species that rebounded after near extirpation. We hypothesized that metabarcoding would detect a greater diversity and frequency of prey, and that notable differences in diet will be detected since prior studies. Grey seal scat samples ( N = 247) were collected between 2018 and 2019 from Monomoy Island, Massachusetts, USA. Metabarcoding detected greater prey richness on average, with more frequent detections of clupeids (Clupeidae) and flatfish (Pleuronectiformes), whereas hard parts analysis more frequently detected phycid hakes ( Urophycis spp. Gill, 1863). Combining methods increased detections of 13 prey taxa, with 32 prey taxa identified overall. Skates (Rajidae), flatfish, clupeids, and sand lance ( Ammodytes spp. Linnaeus, 1758) were top-occurring prey. Our study highlights the importance of using multiple methods to characterize generalist predator diets using non-invasive techniques and suggests grey seal diet has changed since the early 2000s.

Molecular Investigation Using RAPD-PCR Marker Field Populations of Pectinophora gossypiella Saunders (Lepidoptera: Gelechiidae) Exposed to Some Insecticide

Seasonally the cotton plant cultivated in Egyptian fields is suffering from the cotton Pink Bollworm Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) attacking fruits and buds. Insecticide applications reported control failure by the most recommended conventional classes. Monitoring pest DNA changes after sublethal exposure become the new technique to investigate. Then four insecticide treatments were screened by using the molecular marker polymerase chain reaction (PCR) inspection, using four Random amplified polymorphic deoxyribonucleic acid analysis (RAPD-PCR) primers partitioned based on band reproducibility. Result of LC50 was 0.55, 61.1, 69.3, and 0.23 for spinoteram, novaluron, metaflumezone, and dimeuron respectively. The insecticide treatments band of gel produced detected 47 loci ranging from 87 to 63 % polymorphism. The primer efficiency value of PIC =0.361, 0.34, 0.34, 0.355, and 0.262, RP = 5.30, H = 0.473, 0.434, 0.434, 0.462 and 0.31, and MI = 0.10, 0.1386, 0.138, 0.167 and 0.0582 for the same treated samples respectively. Distance and similarity were quantified based on Nei’s and genetic dissimilarity by the UPMGA method then a phylogenetic tree was constructed and grouped the entire genotypes into 2 major clusters and 6 subclusters. The RAPD primers revealed the number of alleles (Na = 0.324, 0.318, 0.318, 0.326, and 0.328), and the effective number of alleles (Ne = 0.571, 0.569, 0.558, 0.574, and 0.5784). The fixation-index (Fst) analysis narrated a very great genetic diversity (Fst = 0.626, 0.684, 0.684, 0.2, and 0.695) exists within the four treated samples respectively. The level of gene flow was (Nm = 0.239, 0.230, 0.230, 0.233, and 0.2187) respectively across the four genotypes studied. Results proved that the RAPD-PCR technique was suitable for distinguishing between insecticide treatments.

Congenital Cytomegalovirus Infection Screening in Newborns From Saliva Samples by Real-Time Polymerase Chain Reaction Analysis.

Congenital cytomegalovirus infection is the most common congenital infection. Although screening of congenital cytomegalovirus infection with polymerase chain reaction studies in blood, urine, and saliva samples has been developed in recent years, it is not yet in routine use in any country. In our study, cytomegalovirus deoxyribonucleic acid analysis was per- formed by real-time polymerase chain reaction method in saliva samples taken before the first feeding during the first day following birth in neonates born in a university hospital between January 2021 and January 2022. To support the diagnosis, additionally, cytomegalovirus deoxy- ribonucleic acid positivity in urine and blood samples was investigated in newborns with cyto- megalovirus deoxyribonucleic acid positivity in saliva. Cytomegalovirus deoxyribonucleic acid was investigated in saliva samples of 545 neonates by real-time polymerase chain reaction method in 1-year period and positiv- ity was found in 6 neonates. Since cytomegalovirus deoxyribonucleic acid was found nega- tive by the real-time polymerase chain reaction method in the urine and blood samples of 5 of these neonates, the positivity in the saliva sample was interpreted as false positivity. In 1 case, cytomegalovirus deoxyribonucleic acid positivity was detected in urine and blood samples 5 weeks later. As a result, definite congenital cytomegalovirus infection could not be diagnosed in 545 cases, while possible congenital cytomegalovirus infection was diag- nosed in 1 case. It has been concluded that the frequency of congenital cytomegalovirus infection is low in our study group and studying saliva samples showed high false-positive rates. It is seen that saliva is not a suitable sample for detecting cytomegalovirus deoxyribonucleic acid by real-time polymerase chain reaction method.

Detection and Management of Freshwater Invasive Alien Species through Environmental DNA Analysis and Geographic Information Systems: A Review

Freshwater invasive alien species (IAS) are non-native organisms that were intentionally or unintentionally released into local water bodies and later harmed the invaded habitat by disrupting the ecological processes. Over the last few years, environmental deoxyribonucleic acid (eDNA) analyses have been used in many studies to detect IAS, with positive results. However, with the help of geographic information systems (GIS), efforts to detect the presence of IAS can be made faster and more efficiently. In this paper, we review the background of IAS in Southeast Asia and management efforts undertaken involving the input of known habitat-specific geographical parameters into GIS mapping. Via this strategy, it is possible to identify and distinguish areas that fit IAS habitat features from those that do not. eDNA analysis can later be applied to confirm the presence of IAS in detected areas, enabling further studies and actions. The presence of IAS in certain areas can be used as an indicator to assess the environmental integrity of native waterways. This combined method is likely the first approach to be applied to the detection of freshwater IAS in local water bodies. Apart from saving energy and resources, embedding GIS and eDNA into the study of IAS not only benefits the ecosystem but also assists locals and authorities in managing and taking necessary enforcement actions to curb further spread.

Application of Next Generation Sequencing (NGS) technology in forensic science: A review

One of the most important achievements of Next Generation Sequencing (NGS) is to produce millions of sequences reads in a short period of time, and to produce large sequences of deoxyribonucleic acid (DNA) in fragments of any size. Libraries can be generated from whole genomes or any DNA or RNA region of interest without the need to know its sequence beforehand. In the forensic field, one of the main problems is the limited amount of sample available, as well as its degraded state samples. Next-generation sequencing (NGS) technology, with its high-throughput capacity and low cost, has developed rapidly in recent years and become an important analytical tool for many genomics researchers. New opportunities in the research domain of the forensic studies emerge by using the power of next generation sequencing technology, which can be applied to simultaneously analyzing multiple loci of forensic interest in different genetic materials. Furthermore, next-generation sequencing (NGS) technology can also have potential applications in many other aspects of forensic science such as achieving the simultaneous analysis of the standard autosomal DNA (STRs and SNPs), deoxyribonucleic acid (DNA) database construction, ancestry and phenotypic inferences, monozygotic twin studies, body fluid and species identification, forensic animal, plant and microbiological analyses, mitochondrial DNA, microbiological analysis, epigenetics analysis, MicroRNA analysis, animal and plant DNA analysis and X and Y chromosomal markers. In this study, we review the application of next generation sequencing (NGS) technology in the field of forensic science with the aim of providing a reference for future forensic studies and practice.

A Rare Case: HaemoglobinS-Thalassemia in Adult

Hemoglobinopathy refers to a disease involving a qualitative or quantitative defect of the structure or synthesis of haemoglobin molecules. The HaemoglobinS- beta thalassemia occurs in a heterozygotes individual with beta-thalassemia and HaemoglobinS gene. A 29-year-old man came with severe anemia, thrombocytopenia, and history of repeated blood transfusions. Physical examination showed pale conjunctiva, pansystolic murmurs, and hepatosplenomegaly. The HaemoglobinS fraction was found in haemoglobin electrophoresis with increased HaemoglobinF and decreased HaemoglobinA2 fraction. The peripheral blood smear shows abnormal erythrocytes morphologies such as pencil shapes, fragmentocytes, target cells, and sickle shapes. The patient was diagnosed with chronic anaemia caused by HaemoglobinS-beta thalassemia. It makes ineffective erythropoiesis, intravascular, and extravascular hemolysis. This haemoglobinopathy caused increased ferritin and transferrin saturation. The presence of renal failure indicate there is a complicated condition like microvascular obstruction of renal. In this case, there is a reduction of HaemoglobinA2 fraction that is not common in HaemoglobinS-beta thalassemia. The patient with Haemoglobin S / beta+ thalassemia shows intravascular hemolysis, ineffective hematopoiesis, and vaso-occlusive signs. Deoxyribo Nucleic Acid analysis is further needed to confirm the combination defect of haemoglobin synthesis disorders in conjunction with alpha thalassemia or Hereditary persistence of fetal haemoglobin.

Circulating Cell-Free Tumor Deoxyribonucleic Acid Analysis as a Tool for the Diagnosis and Monitoring of Pediatric Solid Tumors.

The utility of cell-free tumor deoxyribonucleic acid analysis is currently being evaluated in a wide range of clinical studies. The validity of cell-free tumor deoxyribonucleic acid analysis methods used for screening and detecting malignant diseases, monitoring the effectiveness of treatment and disease progression, and identifying potential relapse is tested. Molecular technologies used for cell-free tumor deoxyribonucleic acid analysis include targeted polymerase chain reaction assays and next-generation sequencing approaches along with newly introduced epigenetic analysis methods such as methylation-specific polymerase chain reaction. The aim of this review was to compare the methods, pitfalls, and advantages of tests developed for the analysis of cell-free tumor deoxyribonucleic acid in the diagnosis and treatment of pediatric solid tumors. For this purpose, the PubMed database was searched for articles published in the last 10 years, in English, that investigated a human cohort aged 0 to 18 years. A total of 272 references were analyzed. A total of 33 studies were included in the review. Cell-free tumor deoxyribonucleic acid analysis has emerged as a promising novel approach that could potentially bring a significant improvement in the field of pediatric oncology, but the implementation of cell-free tumor deoxyribonucleic acid in clinical practice is largely hindered by the lack of standardized methods for processing and analysis.

The Analysis of Genetic Polymorphism on Mitochondrial Hypervariable Region III in Thai Population.

Mitochondrial DNA (mtDNA) analysis is a genetic marker for human identification, especially matrilineal inheritance. Hypervariable regions (HVR) I and II of mtDNA have been currently performed for human identification worldwide. Further examination of HVRIII has been conducted with the aim of enhancing the power of discrimination. The aim of this research is to provide informative data on the polymorphisms of HVRIII in the Thai population in order to establish a national database for human identification. Thai people who were unrelated through the maternal lineage were recruited for blood collections. The mtDNA was extracted by Chelex extraction, amplified by polymerase chain reaction, and analyzed using Sequencing Analysis Software. The most common mutation in HVRIII was base substitution, followed by deletion and insertion. We discovered 40 unique haplotypes, with haplotype 489C being the most frequent. The haplotype diversity, power of discrimination, and random match probability were 0.8014, 0.7987, and 0.2013, respectively. Five-CA repeats were the most frequently observed in nucleotide positions 514-523. Our database can be employed as supplementary markers in addition to nuclear deoxyribonucleic acid (DNA) markers in forensic investigations. Moreover, the data could potentially enhance genetic identification and anthropological genetics research in Thailand.

DNK identifikacija sa posebnim osvrtom na metodu fenotipizacije

Deoxyribonucleic acid (DNA) undeniably plays a unique role in forensic sciences today. Since 1985, when Alec Jeffreys and his colleagues first applied DNA analysis in forensic investigations, numerous cases have been successfully resolved, primarily thanks to this method. It is indisputable that DNA analysis has become a new form of scientific evidence continually interpreted and evaluated by both professionals and the wider public. Increasingly, courts worldwide are accepting the results of DNA analysis, leading to almost universal adoption of this technique in most legal systems. DNA analysis in forensic medicine is employed in investigations of various criminal offenses, determination of identity, and establishing paternity. According to some sources, in the United States alone, over 300,000 DNA analyses are conducted each year. Moreover, several hundred previously convicted individuals, including some on death row, have been exonerated through the application of DNA analysis, as part of the "DNA Innocence" project that has been conducted in the United States for years. The identification of individuals (perpetrators of criminal acts) based on the so-called genetic code obtained from biological material-DNA-has been practiced globally for around two decades.

Quantitative Analysis of Placental Mitochondrial DNA in Severe Hypertensive Pregnancies: A Cross-sectional Study

Introduction: To address the causes of maternal mortality and morbidity, reproductive health and related disabilities is a motto of World Health Organisation (WHO). Gestational hypertension constitutes as one of the main cause for maternal morbidity and mortality. Aetiology of gestational hypertension- remains a mystery. Mitochondrial dysfunction, particularly in trophoblastic cells, could play an important role in causing gestational hypertension and is postulated to be associated with oxidative stress, impaired differentiation and invasion of trophoblastic tissue. Alterations in peripheral blood mitochondrial Deoxyribonucleic Acid (DNA) copy numbers have been thought as a possible biomarker for mitochondrial dysfunction in disorders induced by oxidative stress. Aim: To estimate, evaluate and compare the mitochondrial copy numbers between placenta of severe gestational hypertensive pregnancies and normotensive pregnancies. Materials and Methods: A cross-sectional study was conducted at Tirunelveli Medical College Hospital, Tirunelveli, Tamil Nadu, India, with 25 clinically proven cases of severe gestational hypertensive pregnancies and 25 normotensive pregnancies, for a period of one year (August 2020 to July 2021). The placenta were collected from the Department of Obstetrics, in containers within two hours of delivery and frozen in liquid nitrogen. Relative quantitative analysis of placental mitochondrial DNA (mtDNA), were done by calculating the copy number value by comparing the levels of two nuclear genes viz., BECN1 and NEB to two mitochondrial genes viz., ND1 and ND6 by using real time quantitative Polymerase Chain Reaction (PCR). Results: The placental mitochondrial DNA copy number was higher in severe gestational hypertensive patients than in normotensive pregnancies. A total of 21 (84%) cases of the severe hypertensive placenta had higher copy numbers 100-110 vs only 13 (52%) cases had copy numbers 100-105 in the normotensive group. The difference in copy number value between them was statistically significant with p-value of 0.03. Conclusion: Present study findings suggest significant alterations at the level of the mitochondria in placenta from women with gestational hypertension, which may well contribute to gestational hypertension pathophysiology. Future research should include direct measure of placental and maternal whole body mitochondrial function, and assays that elucidate the potential modifiers of this association.

Multiplexed ISSR Genotyping by Sequencing (MIG-Seq).

Multiplexed inter-simple sequence repeat (ISSR) genotyping by sequencing (MIG-seq) is a simple, rapid, and inexpensive method for detecting single-nucleotide polymorphisms (SNPs) using next-generation sequencing (NGS). The advantages of MIG-seq include easy application to various species without prior genetic information. In addition, this method opens the door to genome-wide nucleotide sequence analyses of low-quality and trace-level deoxyribonucleic acid (DNA) samples, which have previously been difficult to analyze. Another advantage is that the procedure is simple, time-saving, and inexpensive. Recently, MIG-seq has been applied to wild and cultivated plants and has produced novel results. Using invisible DNA information, questions related to gene flow through pollination and seed dispersal, the genetic structure and diversity of populations, clonality, and the hybridization of wild and cultivated plants are being rapidly answered. In this chapter, I present the results of plant research based on MIG-seq and describe the procedure for this method as a user of MIG-seq.

Sex Determination using Primary and Permanent Teeth under Different Environment Conditions: A Deoxyribonucleic Acid-based Study

Abstract Aims The purpose of this study was to determine the sex of primary and permanent tooth pulp tissue under different environmental conditions using deoxyribonucleic acid (DNA) analysis and the polymerase chain reaction (PCR) amplification method. Materials and Methods For this study, 40 permanent and primary teeth (20 males and 20 females) that were scheduled for extraction were used. For 6 months, the teeth were divided into four groups of ten (5 males and 5 females) based on different environmental conditions. The pulp was extracted from each tooth, DNA was isolated using QIAGEN’s DNeasy Blood and Tissue Kit, and sex was determined using a PCR technique. Results DNA was extracted from all 40 samples subjected to various environmental conditions, but sex was correctly determined in only 90% of the samples. Conclusions PCR-based sex determination using primary and permanent teeth under different environmental conditions for up to 6 months is a reliable technique with high accuracy.