Genetic Analysis of SCN5A Gene in Patients of Long QT Syndrome

Background: Finding out the cause of death is the main concern in forensic casework, particularly in sudden unexplained death cases. Long QT syndrome is a form of life debilitating cardiac arrhythmia. Among LQT genes, LQT3 is suggested to be more lethal and patients are at high probability of sudden death as compared to LQT1 and LQT2 patient.Methods: Samples of long QT patients were collected and the coding regions of all exons of the SCN5A, voltage- gated Na+ channel gene were screened by sequencing for the potential mutations as the causative agent of long QT syndrome.Results: After data analysis, 3 genetic variations have been found, amongst them are 2 heterozygous mutations that were reported previously in other ethnicity G87A-A29A found in exon 2, G1673A-H558R in exon 12 and one novel mutation that has not been reported so far, G>A 1238-G412G resulting in a transition mutation and change in amino acid GGG- Glycine to GAG- glutamic acid at position 412 in exon 9. While in other exons, no significant mutation was found.Conclusion: As some of the exons showed mutation and the sample size was small, however, further functional analysis of the gene is needed with large number of samples for the confirmation of results.Keywords: Long QT Syndrome; SCN5A gene; Genetic variations; Sudden deat

Forensic Entomology: A Comprehensive Review

Determination of post mortem interval is done with various methods depending upon the condition of the dead body and the circumstances in which it is found. Ruling out foul play in unnatural deaths has been an enigma of a sort for forensic pathologists and scientists all over the world. The methods to determine the cause and manner of death keep on emerging with ongoing researches in the fields of forensic sciences. Many methods have been devised to determine cause, manner and specifically time since death. Forensic entomology is one such field that aids in determination of time since death, especially in putrefied corpses found in different habitats in mysterious unknown circumstances. The field keeps on emerging with the advent of new techniques on molecular level, including DNA analysis and identification based on entomological data that has diversified over the years. The standards and guidelines for entomological data collection and processing needed for implication on the crime scene must be revised now and again for assistance of crime scene investigators, scientists, entomologists and pathologists. Various factors must be considered while processing entomological data that affect post mortem intervals. The keen study of life cycles, groups of similar insects, valid and standard methods of insect collection, breeding and identification can give a lead on cause, manner, time, place and circumstances of unknown or unnatural deaths.

Overview of the JET preparation for deuterium-tritium operation with the ITER like-wall

For the past several years, the JET scientific programme (Pamela et al 2007 Fusion Eng. Des.82 590) has been engaged in a multi-campaign effort, including experiments in D, H and T, leading up to 2020 and the first experiments with 50%/50% D–T mixtures since 1997 and the first ever D–T plasmas with the ITER mix of plasma-facing component materials. For this purpose, a concerted physics and technology programme was launched with a view to prepare the D–T campaign (DTE2). This paper addresses the key elements developed by the JET programme directly contributing to the D–T preparation. This intense preparation includes the review of the physics basis for the D–T operational scenarios, including the fusion power predictions through first principle and integrated modelling, and the impact of isotopes in the operation and physics of D–T plasmas (thermal and particle transport, high confinement mode (H-mode) access, Be and W erosion, fuel recovery, etc). This effort also requires improving several aspects of plasma operation for DTE2, such as real time control schemes, heat load control, disruption avoidance and a mitigation system (including the installation of a new shattered pellet injector), novel ion cyclotron resonance heating schemes (such as the three-ions scheme), new diagnostics (neutron camera and spectrometer, active Alfvèn eigenmode antennas, neutral gauges, radiation hard imaging systems...) and the calibration of the JET neutron diagnostics at 14 MeV for accurate fusion power measurement. The active preparation of JET for the 2020 D–T campaign provides an incomparable source of information and a basis for the future D–T operation of ITER, and it is also foreseen that a large number of key physics issues will be addressed in support of burning plasmas.This work has been carried out within the framework of the EUROfusion Consortium and has received funding from the Euratom research and training programme 2014–2018 and 2019–2020 under grant agreement No. 633053Postprint (published version

Analysis of mRNA Expression for Injury-Age Estimation

Background: Determination of wound age is pivotal in forensic medical sciences, criminal and civil cases for the construction of crime scene and answering the questions like, time of infliction, manner of wound infliction, how long the person survives after infliction of wound and characterizing antemortem or postmortem wounds. The findings differ considerably among individuals due to the biological variations. Previous developed method in the injury-age determination is clinical, microscopic, enzymatic reaction at the wound margins, histological, and immunohistochemistry with the pitfalls associated with it.Methods: This study is conducted on blunt injuries, particularly lacerated wound (type of wound inflicted by blunt weapon) to analyze the different expression pattern in injury-age up to 72hrs in total 21 individuals randomly grouped in different time intervals. To determine the time of injury, transcript abundance of mRNA of Fibronectin (FBN), IL1β, VEGFA, and GM-CSF was analyzed by real time polymerase chain reaction. 18S-rRNA was used as control marker.Results: Percent knockdown (%KD) was calculated to determine the expression of mRNA for the determination of injury-age. IL1β and GM-CSF showed the predictive behavior for wound age up to 36hrs, Fibronectin (FBN) showed predictive behavior up to 12hrs while VEGFA showed prediction beyond 72hrs.Conclusion: The detection of gradual decrease of mRNA of Fibronectin (FBN), IL1β, VEGFA, and GM-CSF may provide an estimation of wound-age.Keywords: mRNA Expression; Injury-Age Estimation; Antemortem wounds; Postmortem wound

DNA extraction and quantification from human saliva deposited on fruits with human bite

Background: Among important aspects of forensic science there stand recovery, preservation and analysis of stains originated by body fluid. DNA Isolated from evidence stains help to exclude an innocent suspect or to identify a perpetrator upon PCR-based typing. This study reports extraction and quantification of DNA from human saliva deposited on fruits. The research work was conducted at Department of Forensic Sciences, University of Health Sciences, Lahore and WTO laboratory of University of Veterinary and Animal Sciences, Lahore.Methodology: DNA from saliva deposited on bitten fruits. DNA from 55 samples were extracted by Chelax method, Quantifiler® DNA Quantification Kit was used for quantification of the extracted samples and amplification of DNA was done in 7500 Real-Time PCR systems (Applied Biosystems, Foster City, CA). Total 55 samples including controls (positive and negative controls) were collected from bitten fruits by sterile swabs.Results: All sample swabs with human saliva showed result for quantification. Overall good yield of DNA quantification obtained from all fruits and no sample showed internal inhibition. No sample showed non informative or incomplete quantification which occurs due to LCN (Low copy number) or lesser quantity of DNA extracted from saliva swab.Conclusion: This study has provided with optimized protocol to isolate DNA from saliva found in very minute quantity on organic surfaces like fruits. Adaptation of this method can play a vital role in establishing new trends in human forensics practiced in Pakistan. Moreover, for future work in human forensics, this study can provide important practical basis

Population Density, Habitat Characteristics and Preferences of Red Fox (Vulpes vulpes) in Chakwal, Pakistan

The Red fox (Vulpes vulpes) is a least concern carnivore according to the IUCN Red List of Threatened Species (2016). However, in Pakistan Red fox is considered as Near Threatened (NT), due to habitat destruction and depletion of food resources. The objective of the study was to identify habitat preferences and population density of Red fox in District Chakwal, Pakistan. Line transect census method was used to estimate the population density of Red fox through direct sighting and indirect method of burrow counting, presence of footprints and scats. A total of 10 transects were carried out at three potential sites: Devi, Photaki and Chumbisurla Wildlife Sanctuary (CWS) in Chakwal based on preliminary surveys. Habitat preference was estimated by comparing three different study sites by quadrat method and found that CWS area is preferred habitat for Red fox. A total of 24 plant species were recorded in the study areas, among them Cynodon dactylon is major herb found to provide shelter to Red fox in all study sites based on Importance value Index (I.V.I) at CWS (IVI=208.8) followed by Devi (IVI=185.93) and Photaki (IVI=142.33). The maximum population density of Red fox through direct sighting at CWS having 0.26 individuals/km2 compared to Devi and Photaki having 0.16 and 0.13 individuals/km2, respectively. The indirect estimation method revealed that maximum dens were found in CWS area compared to Devi and Photaki, while footprints and scats were found maximum in Devi and Photaki, respectively. It is concluded that Red fox preferred habitat is CWS site. Habitat destruction and conflicts with fox are causing the population of the Red fox to dwindle in Chakwal, Pakistan

In Vitro Supplementation of Glycine Improves Quality of Cryopreserved Bull Semen

The extender supplementation of glycine on quality of cryopreserved bull sperm was reported in current study. Three adult Sahiwal bulls was used for semen collection by artificial vagina having 42°C (three weeks) (replicates; n=18)). Collected semen was quickly shifted to laboratory for initial evaluation. Qualified semen ejaculates (>60% motility, >1mL volume, >0.5billion/mL cons.) was diluted in tris extender having 0 (control), 5, 10 and 15 and 20mM of glycine. Cooling of extended semen to (4℃) was done in 2 hours and kept for 4 hours for equilibration. French straws were used for filling at (4℃), liquid nitrogen used for freezing. Highest percentages of post thaw sperm progressive motility and acrosome integrity were observed in extender containing 5mM of glycine compared to other treatments and control (P<0.05). However, glycine supplementation in extender didn’t affect viability, chromatin integrity and plasma membrane integrity of bull sperm. In conclusion, glycine addition at 5 mM in extender improves quality of cryopreserved bull semen

Are Roma People Descended from the Punjab Region of Pakistan: A Y-Chromosomal Perspective

Gypsies are a separate ethnic group living in Pakistan and some other countries as well. They are mostly known as ‘Roma’ and ‘untouchables’. They have different types of lifestyles as compared to other common people, as they always keep migrating from one place to another. They do not have proper houses; they live in tent houses and most probably work on daily wages to earn their living. Gypsies cannot be specified according to the place of residence and can only be classified according to their migration route. Previous historical and linguistic research showed the north Indian origin of Roma people. The present study collected 285 unrelated Roma individuals living in Punjab and typed with the Goldeneye Y20 system. Allelic frequencies ranged between 0.0035 and 0.5266, with haplotype diversity (HD) of 0.9999 and discrimination capacity (DC) of 0.8790. Gene diversity (GD) ranged from 0.6489 (DYS391) to 0.9764 (DYS391) (DY385ab). A total of 223 unique alleles were observed. Interestingly, the haplogroup R accounted for 40.56% and J for 22.06%. In MDS analysis, Pakistani Roma formed a close cluster with Roma from Constanta, Romania. The migration pattern of the Roma population from Pakistan, India and Europe was inferred using coalescence theory in the Migrate-n program. Overlapping Y-STR data were used to test different migration models. These migration models showed us the dominant gene flow from Pakistan to India and Europe to Pakistan. The results of our study showed that Y STRs provided substantially stronger discriminatory power in the Pakistani Roma population

Simulations of Alfvénic modes in TJ-II Stellarator

Alfvénic modes are one of the subclass of instabilities prevalent in burning plasmas due to interaction of energetic particles with background plasma. In this paper we investigate the properties of these modes with 3D simulations using modeling tools STELLGAP [1] and AE3D [2] of Neutral Beam Injection (NBI) heated H-plasmas in TJ-II low-magnetic-shear flexible heliac (B0 = 0.95 T, = 1.5 m, = 0.22 m). These simulations focus on modelling the experimental observations [3] for prominent modes in TJ-II plasmas. Our simulations show consistency in frequency and radial location with the measured Alfvén Eigenmodes [3]. These simulations are performed for chirping and steady modes in TJ-II discharge # 29839 at t = 1150 and 1160ms respectively