Parkash Chand

Assessment of DNA damage and repair efficiency in drug naïve schizophrenia using comet assay

The etiology of schizophrenia continues to be confounding and elusive. Some knowledge gaps exist in the neurodegenerative theory of schizophrenia. Oxidative DNA damage and repair deficits are relevant to the mechanisms of neurodegeneration but have not been studied in drug naïve schizophrenia. The present study used the comet assay technique to study the extent of DNA damage in circulating peripheral lymphocytes of patients with drug naïve schizophrenia (n = 40) along with an age and gender matched control group (n = 40). We also assessed the DNA repair efficiency in cases following incubation in a nutrient medium. All the assayed comet parameters demonstrated significantly greater baseline DNA damage in cases in comparison to the controls except for head diameter (p < 0.001 for all significant results, p = 0.32 for head diameter). Gender, age and duration of illness (p = 0.21, 0.69 and 0.12 respectively for tail length) did not influence any of the parameters significantly. Significant decrease was noted in the comet tail length and percentage of DNA in comet tail (p < 0.001 for both) in cases following incubation suggesting that the DNA repair machinery was preserved. No difference in DNA repair efficiency was noted between the genders (p = 0.23 for tail length). Our findings confirm the presence of significant baseline DNA damage in schizophrenia even prior to the initiation of anti-psychotic treatment. Additionally, intact genomic repair efficiency was noted in this group as a whole. These results provide some evidence for oxidative DNA damage as molecular link underpinning neurodegeneration in drug naïve schizophrenia.

Assessment of folic acid and DNA damage in cleft lip and cleft palate

Studies have identified the risk factors like folic acid deficiency during gestational period, family history for orofacial clefts, drugs like antiepileptic, vitamin A. But, the data regarding the folic acid status in children with cleft lip/palate is hardly evaluated in depth. Here, an assessment of folic acid and DNA damage were carried out in children with orofacial anomalies. Folic acid level and DNA damage were evaluated by folic acid assay (direct chemiluminescent technology) and single cell gel electrophoresis or comet assay method respectively. The mean value of plasma folic acid by direct chemiluminescent technology was 6.5±3.6 nmol/L and the normal value in children ranges from 11.3 to 47.6 nmol/L. The amount of damaged DNA, measured as the tail length of the comet in cases, was 19.4±8.9 μm and the mean percentage of DNA in tail was 16.5±3.7. Folic acid deficiency could be the reason for DNA damage.

Automatic detection of comets in Silver stained comet assay images for DNA damage analysis

Comet assay or Single Cell Gell Electrophoresis (SCGE) is a standard method for detecting Deoxyribo Nucleic Acid (DNA) damage in individual cells. The final stage of comet assay process is image acquisition and analysis. An effective automatic method for detecting comets from very noisy comet assay images is proposed in this paper. Most of the related work in this area are based on fluorescent stained comet assay images. But, silver staining method is commonly used for clinical applications in developing countries like India as it is inexpensive, slides can be preserved for a long time and is less hazardous. Currently most of the comet assay systems employ image analysis modules that are semi-automated which have the disadvantages of requirement of skilled personnel and having limited throughput. Hence, it is evident that a fully automated image analysis system is critical for rapid and accurate DNA damage detection. Therefore, an automatic comet detection stage as part of a fully automated comet assay image analysis system for DNA damage analysis and quantification is developed and implemented here. Based on our results and evaluation by a clinical expert, the performance of the proposed method is found to be better than that of the existing methods in terms of %TP, %FN, %FP, PPV and sensitivity.

Assessment of sperm deoxyribose nucleic acid fragmentation using sperm chromatin dispersion assay

The integrity of sperm deoxyribose nucleic acid (DNA) is one of the determinants that ensure normal fertilization, implantation, pregnancy, and the generation of a healthy progeny. Assessment of sperm DNA fragmentation has gained importance as a tool to provide significant information regarding sperm quality, and it can independently predict sperm fertilizing potential. The sperm chromatin dispersion (SCD) assay is one of the available techniques to detect sperm DNA damage with results comparable to the gold standard – Sperm Chromatin Structure Assay. We present here a detailed methodology of an alkaline modification of SCD that can be carried out with minimal laboratory equipment. The SCD assay is a modified halo assay that utilizes chemical methods to demonstrate sperm DNA fragmentation. It involves the embedding of sperms in an agarose medium followed by exposure to alkaline denaturation and deproteinization. The results are interpreted under a light microscope. A visual scoring system is utilized to differentiate the sperms with possible DNA fragmentation from those without fragmented DNA and to determine the sperm DNA fragmentation index (SDFI) % for each semen sample. The SDFI % is directly proportional to the quality of sperm. The SCD assay is a simple, cost-effective, and reliable technique that can detect sperm DNA fragmentation, thus providing information regarding sperm functional quality and reproductive capacity. It is of significance in clinical and research areas of andrology and reproductive medicine, toxicology, and pharmacotherapeutics.

Variations in the branching pattern of posterior division of mandibular nerve: a case report

PurposeAbnormal communications among the branches of mandibular nerve especially the posterior division are significant due to various procedures undertaken in this region. These variations are worth reporting as they pose serious implications in several interventions in this region, and may even lead to false diagnosis.MethodsDuring routine dissection, the mandibular nerve and its branches were dissected in the infratemporal fossa. The branches from the posterior division of the mandibular nerve namely the inferior alveolar and auriculotemporal nerves were carefully dissected, and their abnormal branching pattern was noted.ResultsThere was a communicating branch between left inferior alveolar and auriculotemporal nerve. There was also a variant recurrent branch from the left inferior alveolar nerve that supplied the lateral pterygoid muscle.ConclusionsSuch variant branches and communications between the branches of mandibular nerve as seen in this case have an embryological basis and are clinically important in this region especially for dental surgeries and anesthesia.

Oxidative stress and DNA damage in newborns born to mothers with hyperglycemia – a prospective cohort study

Abstract Background: Hyperglycemic mothers have increased oxidative stress during pregnancy, which can adversely affect the outcome in their neonates. Objectives: To measure the oxidative stress and DNA damage in newborns born to mothers with hyperglycemia and correlate their immediate outcome with the amount of oxidative stress and DNA damage. Methods: This prospective cohort study conducted in a tertiary care teaching hospital, South India included three groups – newborns born to pregestational diabetic mothers, gestational diabetic mothers, and euglycemic mothers with 24 newborns in each. Oxidative stress parameters – malondialdehyde (MDA), total antioxidant capacity (TAC), and DNA damage (comet assay) were assessed in umbilical cord blood. Association between these parameters and immediate neonatal outcome among three groups were studied. Results: Newborns born to diabetic mothers had more MDA levels and DNA damage compared to euglycemic mothers. There was a positive correlation between comet parameters and MDA levels. There was an inverse correlation between comet parameters and TAC. Newborns born to diabetic mothers were at high risk to develop respiratory distress syndrome, hypoglycemia, and sepsis. Conclusions: Newborns born to diabetic mothers had increased oxidative stress and DNA damage compared to those born to mothers with euglycemia.

A rare musculo-septal aponeurotic tunnel in the arm with anomalous arterial pattern

Aparna Muraleedharan , Raveendranath Veeramani *, Parkash Chand c Assistant Professor (Anatomy), Pondicherry Institute of Medical Sciences, Ganapathichettikulam, Kalapet, Puducherry 605014, India Assistant Professor (Anatomy), Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Dhanvantri Nagar, Puducherry 605006, India Professor & Head (Anatomy), Jawaharlal Institute of Postgraduate Medical Education & Research (JIPMER), Dhanvantri Nagar, Puducherry 605006, India m e d i c a l j o u r n a l a r m e d f o r c e s i n d i a x x x ( 2 0 1 6 ) x x x – x x x

Body Donation- Consent from Non-Related Persons: Case Series, Review, and Recommendations

Learning anatomy by dissection of cadavers is the best way to learn anatomy. Voluntary body donation is one of the sources of procuring cadavers. In the case of donations after hospital or non-institutional deaths, the family members of the deceased approach the hospital authorities regarding body donation of the deceased. There are situations, where there is no available accompanying near relatives of the deceased, which pose a challenge for personnel involved in the process of body donation. In two of the reported cases, the body donation was done by the live-in partner and a friend of the deceased. In another reported case, the son of the deceased was nominated by the donor at the time of registration. As the son of the deceased was not available at the time of death of the donor, donation was executed by another near relative. Anatomy Acts of individual States in the Union of India and Acts of other countries are examined about the consent for body donation by persons other than near relatives. None of the Anatomy Acts of various States in India provide an alternative in the absence of near relatives for claiming the body or for donating the body, except for Acts of Delhi and Kerala. There is a need to bring in a Unified Anatomy Act, common for all the States and Union territories in India and include: friend, live-in partner and a nominated person in the provisions of the Act to enable them to give consent for body donation.

DNA Damage Analysis in Children with Non-syndromic Developmental Delay by Comet Assay.

INTRODUCTION Majority of the developmental delays in children are non-syndromic and they are believed to have an underlying DNA damage, though not well substantiated. Hence the present study was carried out to find out if there is any increased DNA damage in children with non-syndromic developmental delay by using the comet assay. AIM The present case-control study was undertaken to assess the level of DNA damage in children with non syndromic developmental delay and compare the same with that of age and sex matched controls using submarine gel electrophoresis (Comet Assay). MATERIALS AND METHODS The blood from clinically diagnosed children with non syndromic developmental delay and controls were subjected for alkaline version of comet assay - Single cell gel electrophoresis using lymphocytes isolated from the peripheral blood. The comets were observed under a bright field microscope; photocaptured and scored using the Image J image quantification software. Comet parameters were compared between the cases and controls and statistical analysis and interpretation of results was done using the statistical software SPSS version 20. RESULTS The mean comet tail length in cases and control was 20.77+7.659μm and 08.97+4.398μm respectively which was statistically significant (p<0.001). Other comet parameters like total comet length and % DNA in tail also showed a statistically significant difference (p < 0.001) between cases and controls. CONCLUSION The current investigation unraveled increased levels of DNA damage in children with non syndromic developmental delay when compared to the controls.

Common peroneal nerve entrapment with the communication between inferior gluteal nerve and common peroneal nerve

Sciatic nerve divides into tibial nerve and common peroneal nerve at the level of superior angle of popliteal fossa and variations in its branching pattern are common. The most common nerve entrapment syndrome in the lower limbs is common peroneal nerve entrapment at fibular head. Invariably it can also be trapped in gluteal region due to split piriformis muscle which compresses sciatic nerve as whole or its division such as common peroneal nerve. In this case report, we describe a case of high division of sciatic nerve along with duplicated piriformis associated with a communicating branch between inferior gluteal nerve and common peroneal nerve. Such an anatomical variation of common peroneal nerve with the communicating nerve is discussed with its embryological basis.