Alterations of oligodendrocyte progenitor cells (OPCs) with survival time in humans following high impact brain trauma.

BACKGROUND: As there is a lack of comprehensive literature regarding the molecular environment of the human brain emphasizing on oligodendrocyte progenitor cells (OPCs) following high impact brain trauma. The protagonist of OPCs post severe traumatic brain injury (sTBI) provides a significant thrust towards estimating time elapsed since trauma as well as developing novel therapeutic approaches. The present study was carried out to study post trauma alterations pertaining to myelin sheath and oligodendrocyte response with survival time. MATERIALS AND METHODS: In the present study, victims (both male and female) of sTBI (n = 64) were recruited and contrasted with age and gender matched controls (n = 12). Post mortem brain samples from corpus callosum and grey white matter interface were collected during autopsy examination. Extent of myelin degradation and response of OPC markers Olig-2 and PDGFR-α were evaluated using immunohistochemistry and qRT-PCR. STATA 14.0 statistical software was used for data analysis with P-value<0.05 considered statistically significant. RESULTS: Timewise qualitative correlation with extent of demyelination performed using LFB-PAS/IHC-MBP, IHC Olig-2 and mRNA expression revealed tendency towards remyelination in both corpus callosum and grey white matter interface. Number of Olig-2 positive cells was significantly higher in sTBI group as compared to control group (P-value: 0.0001). Moreover, mRNA expression studies of Olig-2 showed significant upregulation in sTBI patients. mRNA expression of Olig-2 and PDGFR-α in sTBI patients showed significant variation with respect to survival time (p value:0.0001). CONCLUSION: Detailed assessment of post TBI changes implementing various immunohistochemical and molecular techniques shall potentially reveal intriguing and important inferences in medicolegal practices and neurotherapeutics.

Chimeric status of biological samples after HSCT for personal identification: Y-STR based DNA analysis in sex mismatch cases.

Identification of an individual is the prime object in forensic case works both in civil or criminal situations like paternity/maternity disputes, sexual assaults, murder, mass disaster victims etc. STR analysis has already proved its potential to give accurate results. In addition to autosomal chromosomes, sex determination at many times is crucial in forensic situations, especially in situations like rape cases or in cases of missing persons. The chances of wrong interpretations may arise due to false detection (or non-detection) of STR fragments overall or only at amelogenin-specific fragments, in situation like mutations, intersex conditions, trans-sexualism etc., due to natural or artificial chimersim. The forensic relevance of the possible misinterpretation of STR's or amelogenin should never be underestimated. The present study was carried out to identify an individual using Y-STR in sex mismatch patients who received hematopoietic stem cell transplantation. Hematopoietic stem cell transplantation is a method to replace patient's stem cell with the stem cell donated by the donor preferably biological related in order to cure malignant and non malignant diseases. This study enrolled ten female patients of HSCT. Samples were collected as pre and post transplant after 15 days, 30 days, 60 days, and 90 days of time interval from sex mismatch patient (female) and from donor (male) and chimeric status of the patient was analyzed using Y-STR markers (23 loci). Results demonstrated that donor genotype existed in blood and buccal swab of the recipient but no genetic profile existed for Y-STR in hair follicle of the recipient. This study suggests that only hair follicle out of three biological samples tested serves as reliable source of recipient's origin after HSCT for accurate personal identification especially in forensic situations.

Benefit of STR-based chimerism analysis to identify TA-GVHD as a cause of death: Utility of various biological specimens.

Transfusion-associated graft-versus-host disease (TA-GVHD) is a rare condition. It can occur after blood transfusion in immune-compromised and occasionally even in immune-competent patients, and is associated with a mortality rate of >90%. The diagnosis of TA-GVHD is often delayed because of its non-specific clinical features. A case of an immune-competent child who developed TA-GVHD is reported here. DNA profiling (short tandem repeat analysis), a technique that has a wide application in forensic medicine, was performed to detect the presence of donor cells in this patient. The findings suggest that more studies are needed with this tool, and the diagnostic potential of using other multiple biological specimens for DNA profiling such as the hair follicle and buccal swab should be evaluated. This is the first case report where the donor's DNA fingerprinting pattern was substantiated from a patient's hair follicle sample. Chimerism was also present in the blood and buccal swab specimens.

Genetic and Cultural Reconstruction of the Migration of an Ancient Lineage.

A rare R1a1 Y-haplogroup (Y-HG) L657 clade subtype designated as LPKSTR is found in most male members of a clan of "founder" families within the Goud Saraswat Brahmin community in Lotli town in Western India. TMRCA calculations using pairwise comparisons to control cohorts suggested a probable migration history distinct from the canonical narrative for medieval migration of orthodox Brahmin families to South India. Using Y-HG centroid analysis, chi-square analysis of TMRCA distributions and archeological find-spots, and discriminant function analysis we show that the parental Z93 L342.2 subclade in which LPKSTR occurs originated in West Asia and that LPKSTR individuals migrated toward the southeast by a Bolan Pass route distinct from the traditionally presumed route of Brahmin ingress into the Indian subcontinent. The proposed migration route is supported by archeological, toponymic, numismatic, linguistic, iconographic, and literary data. Lastly, we present cultural metrics demonstrating that these LPKSTR lineages retained distinct family practices with respect to literacy, religious practice, and emigration not shared with orthodox Brahmins of canonical geographic origin within the same community, despite centuries of intermarriage. Long-term transmission of differentiated family practices within a patrilineal endogamous community has rarely been documented.

Evaluation of blood, buccal swabs, and hair follicles for DNA profiling technique using STR markers.

AIM: To study the short tandem repeat (STR) pattern of DNA from the blood, buccal swabs, and hair follicles of the recipients of allogenic hematopoietic stem cell transplantation to examine whether these tissues contain donor derived cells. METHODS: The study enrolled 25 patients who sustained engraftment. Peripheral blood, buccal swabs, and hair follicles were collected on days 21-30, 90, and 180 after transplantation and the chimeric status of the recipients was evaluated. RESULTS: Donor derived cells existed in the blood and buccal swabs, but not in hair follicles, which can be used to obtain the pre-transplant sample of the recipient after transplant. CONCLUSION: Peripheral blood and buccal swab do not serve as a reliable source of recipient's origin for DNA analysis of individuals who underwent allogenic hematopoietic stem cell transplantation at least within 6 months after transplant.

Effect of acute exposure of triazophos on oxidative stress and histopathological alterations in liver, kidney and brain of Wistar rats.

Acute dose of organophosphorus pesticide Triazophos (O,O-diethyl O-1-phenyl-1H-1,2,4-triazol-3-yl phosphorothioate; Tz) administered orally affects oxidative stress parameters and the histo-architecture of liver, kidney and brain tissues. The results indicate a dose dependent induction of oxidative stress as evident by increased malondialdehyde level and decreased antioxidant defense including glutathione and superoxide dismutase activity in rat liver, kidney and brain. AChE activity was found significantly decreased in the Tz treated groups as compared to the vehicle control (DMSO) group. Histopathological examination of liver, kidney and brain in Tz treated rats revealed medullary congestion and hydropic degeneration of hepatocytes in liver and medullary congestion in kidney. However, no significant histopathological changes were observed in brain tissues.

Quality and quantity of extracted deoxyribonucleic Acid (DNA) from preserved soft tissues of putrefied unidentifiable human corpse.

CONTEXT: The appropriate collection and preservation of soft tissues from putrefied unidentifiable human corpse for the purpose of identification using DNA profiling technique is critically important especially in developing countries like India having different levels of health-care set ups with largely varying facilities and varying climatic conditions. AIMS: The present study was carried out, mainly focusing on quality and quantity of extracted DNA from the soft tissues of putrefied unidentifiable human corpse stored upto 4 weeks at 4°C and at -80°C for DNA analysis. MATERIALS AND METHODS: The present study was conducted on 16 different putrefied unidentifiable human corpses after getting approval from institutional ethical committee. Around 2 g of four different tissues (brain, kidney, heart and muscle) were collected and preserved for one month followed by DNA extraction using the organic method, the quality and quantity of high molecular weight-DNA was estimated using the spectrophotometer and gel electrophoresis. Further, the amplification polymerase chain reaction (PCR) was also performed (AmpFLSTR(®) Indentifiler™ PCR Amplification kit for multiple loci, of Applied Biosystems, Lab India) and was checked using continuous PAGE. RESULTS: The yield of DNA was significantly higher at -80°C for all the four tissues collected and was best for brain followed by heart, kidney and worst for muscles in all cases. CONCLUSIONS: It is suggested that the brain tissue preserved at -80°C is the best among soft issues for DNA extraction. Refrigeration or deep freezing facility should be available at all the centers.

Determination of median lethal dose of combination of endosulfan and cypermethrin in wistar rat.

The present study was designed to determine the lethal dose 50 (LD50) of combination of cypermethrin, a pyrethroid, and endosulfan, an organochlorine compound in Wistar rats. LD50 is the amount (dose) of a chemical, calculated as per the concentration of chemicals that produces death in 50% of a population of test animals to which it is administered by any of a variety of methods. A single oral dose of combination of cypermethrin and endosulfan were dissolved in dimethyl sulfoxide (DMSO) in a ratio of 1:1 and administered orally at the concentration of 165 mg/kg body weight (b.w), 330 mg/kg b.w, 660 mg/kg b.w, and 1320 mg/kg b.w to experimental animals. LD50 was calculated according to the method described by Miller and Tainter (1994) and was observed as 691.83 mg/kg b.w for this combination. Single dose of test article at 165 mg/kg b.w did not reveal any toxic signs or behavioral alterations, hence considered as No observed Adverse Effect level (NOAEL).

Acute oral toxicity and histopathological study of combination of endosulfan and cypermethrin in wistar rats.

BACKGROUND: Endosulfan, a neurotoxic organochlorine insecticide and cypermethrin, a synthetic pyrethroid insecticide used to control pests in domestic, industrial, and agricultural situations. MATERIALS AND METHODS: The present study was carried out to investigate the acute oral toxicity, behavioral and histopathological changes of combination of endosulfan and cypermethrin in albino rats. According to Miller and Tainter analysis method, at 48 h, LD50 value of combination of endosulfan and cypermethrin (ratio 1:1) in rats was found to be 691.83 mg/kg bw by oral gavage. RESULTS: When combination of both these pesticides was administered orally at concentration of 103.72 mg/kg bw, 172.95 mg/kg bw and 207.50 mg/kg bw, respectively, as a single dose, no significant changes in behavior of rats was observed, neither in dosed nor in control group of rats. Combination of endosulfan- and cypermethrin-treated rats showed mild histopathological changes in liver and kidney in group IV (207.50 mg/kg BW) as compared to the control. However, no significant changes were observed in brain and small intestine at either dose of combination of endosulfan and cypermethrin with respect to control. CONCLUSION: Thus, the present study, first of its kind in India, demonstrated the oral toxicity, behavioral, and histo-architectual alterations after induction of combination of endosulfan and cypermethrin at acute doses in Wistar rats.

A psychological profile of a serial killer: a case report.

Serial killers have always fascinated society. A serial killer is typically defined as a perpetrator who murders three or more people over a period of time. Most reported cases of serial killers come from the United States and Canada. In India, there are few reported cases. We present, to the best of our knowledge, the first Indian case in the literature. The present case is of a 28-year-old man, Surinder Koli. The Department of Forensic Medicine & Toxicology, All India Institute of Medical Sciences, New Delphi handled the forensic study. We present a most unique psychological investigation into the mind of a serial killer.

Identification of tumor specimens by DNA analysis in a case of histocytological paraffin tissue block swapping.

We report on a patient who was diagnosed with high-grade breast carcinoma by all the pre-surgery clinical evidence of malignancy, but histopathological reports did not reveal any such tumor residue in the post-surgical tissue block. This raised a suspicion that either exchange of block, labeling error, or a technical error took place during gross examination of the tissue. The mastectomy residue was unprocurable to sort out the problem. So, two doubtful paraffin blocks were sent for DNA fingerprinting analysis. The partial DNA profiles (8-9/15 loci) were obtained from histocytological blocks. The random matching probability for both the paraffin blocks and the patient's blood were found to be 1 in 4.43E4, 1.89E6, and 8.83E13, respectively for Asian population. Multiplex short tandem repeat analysis applied in this case determined that the cause of tumor absence was an error in gross examination of the post-surgical tissue. Moreover, the analysis helped in justifying the therapy given to the patient. Thus, with DNA fingerprinting technique, it was concluded that there was no exchange of the blocks between the two patients operated on the same day and the treatment given to the concerned patient was in the right direction.

Haplotype diversity of 17 Y-chromosomal STRs in Saraswat Brahmin Community of North India.

In this study, 17 Y-specific STR loci (DYS19, DYS389I, DS389II, DYS390, DYS391, DYS392, DYS393, DYS385a/b, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635 and Y_GATA_H4) were analyzed in 181 unrelated male individuals from three North Indian states. A total of 157 different 17-loci haplotypes were identified, 145 of which were unique. The most frequent haplotype was detected in nine instances, occurring with a frequency of 4.97%. These results, including the haplotype data at 17 Y-STR loci in the present study, provide useful information for forensic practice in the Saraswat Brahmin population in North India.

Misinterpretation of results in medico-legal cases due to microdeletion in the Y-chromosome.

In an alleged rape case, for one male suspect, XX genotype and deletion at four Y-STR loci was noticed. The expressions of 18 Y-linked genes were studied to measure the extent of deletion. No expressions at two loci were observed that might have caused the misinterpretations in forensic casework.

Genetic polymorphisms for 17 Y-chromosomal STR haplotypes in Jammu and Kashmir Saraswat Brahmin population.

In this study 17 Y-chromosomal STRs (including DYS19, DYS389I, DS389II, DYS390, DYS391, DYS392, DYS393, DYS385a/b, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635 and Y GATA H4) were analysed using blood samples of 122 unrelated male individuals belonging to Saraswat Brahmin community from Jammu (ID YP000599) and Kashmir (ID YP000600) region of J&K state of India. The allelic frequency distribution and haplotype diversity of 17 Y-chromosomal STR for both the populations were calculated. In the Kashmiri Saraswat group, a total of 109 haplotypes were identified in 122 individuals, of these haplotypes, 101 were found only once. The gene diversity values of STR loci ranged from 0.4813 (DYS391) to 0.8645 (DYS385a/b) for Jammu & Kashmiri Saraswat Brahmins.

Identity of victims from fragmented and decomposed remnants by DNA profiling in a case of serial killings.

A 28-year-old man, Surinder Koli, from a Nithari village adjoining Delhi, India committed serial murder. This case was of paramount importance in medico-legal investigations, as it was a landmark case of a serial killer reported from India. The skeletal remains (627 pieces) including skull/skull portions (19) were recovered from the nearby sewer drain, sump and the backyard of the house in which this man was residing. In addition, soft tissues (51) were also recovered from the same sewer drain. The victims were killed over a two-year period. The establishment of identity of the victims was crucial to prove the case in the court of law as well as for the claimants. Nineteen sets were prepared by radiology/anatomical examination from the exhibits recovered. DNA profiling confirmed the correctness of these sets and also short tandem repeat typing of nuclear DNA successfully identified eight individuals. Both DNA profiling and radiography/anatomical examination played an important role in solving this complicated case.

Validation of qualitative test for phosphine gas in human tissues.

Phosphine has been known to science since the birth of modern chemistry. WHO reports that the technical product usually has a foul odour, like "fishy" or "garlicky" because of the presence of substituted phosphines and diphosphine (P2H4). Many medico-legal autopsy cases have been reported positive for aluminium phosphide even though there was neither any suspicion of consuming aluminium phosphide nor any clinical findings, postmortem findings or circumstantial evidences. The present study was carried out to validate the qualitative test, presently applied in many laboratories for testing phosphine. It was observed that 65% of human tissues in saturated solution of common salt show positivity for phosphine gas on the first day of autopsy.

Application of DNA fingerprinting in medicolegal practice.

Fingerprinting is thought to establish the identify of an individual in forensic cases. The technique is extensively used for forensic purposes. Deoxyribonucleic acid (DNA) is the vehicle of generational transference of heritable unit. While arching markers for genetic disease professor Alec Jeffreys discovered that certain regions of DNA showed variations in the number of tandem repeats known as variable number of tandem repeats (VNTRs). Thus DNA fingerprint was named by observing the number of repeated sequences which differ from individual to individual. The structure of DNA is quite flexible, within the nucleus of each cell resides an identical copy of the individual's genetic material, DNA. The coding regions of the genomic DNA are known as genes. The DNA fingerprinting in forensic science has generated considerable excitement in the criminal justice community. DNA fingerprinting can be applied to identify an individual in criminal and civil cases. Polymerase chain reaction has revolutionised molecular biology it has an ability to amplify (usually fewer than 3000 bp) a particular sequence of DNA into million of copies in a very short period. Consequently only a very tiny amount of an organism's DNA needs to be available originally. This property of polymerase chain reaction has enabled to analyse many forensic samples particularly which are degraded. Microsatellite DNA or commonly as short tandem repeats are scattered throughout the human genome and occur on an average of every 10,000 nucleotides. Microsatellite markers are considered to be the most powerful genetic markers. Collection, preservation and handling are the integral part of DNA fingerprinting analysis. There are various methods to isolate DNA from different biological materials but presently most of the laboratories prefer using FTA paper. The age of humans can be estimated by using DNA based on telomere shortening.