Kuldeep Mohanty

Screening of the LTBP2 gene in a north Indian population with primary congenital glaucoma

Propolis is a potential source of natural antioxidants such as phenolic acids and flavonoids. Its wide biological effects have been known and used since antiquity. In the modern world natural substances are sought which would be able to counteract the effects of antioxidative stress, which underlies many diseases, such as cancer, diabetes and atherosclerosis. This paper aims to present the antioxidative activity of phenolic acids and flavonoids present in Polish propolis and the relationship between their chemical structure and antioxidative activity influencing its medicinal properties. Data concerning the biological activity of propolis are summarized here, including its antibacterial, anti-inflammatory, anticarcinogenic, antiatherogenic, estrogenic effects, as well as AIDS- counteracting and reparative-regenerative function.

Yoga and Meditation as a Therapeutic Intervention in Oxidative Stress and Oxidative DNA Damage to Paternal Genome

Yoga and meditation (encompassing physical postures, breathing practices, relaxation techniques and meditation) is known to modulate neural, endocrine and immune functions at the cellular level through influencing cell cycle control, aging, oxidative stress (OS), apoptosis and several pathways of stress signalling. Individuals with high level of oxidative stress are in great need of safe, inexpensive, non-pharmacologic, accessible, and effective adjunctive therapies to enhance well-being, reduce the burden of such stress and prevent its chronic sequelae. OS is the aetiology in several complex lifestyle diseases (coronary artery disease, hypertension, primary open angle glaucoma, idiopathic male infertility) and autoimmune disorders. Seminal oxidative stress and sperm DNA damage may be the common underlying cause for infertility, recurrent spontaneous abortions, congenital malformations and complex neuropsychiatric disorders in children and in certain childhood cancers. Thus management by antioxidants and simple life style modifications and interventions like meditation and yoga are highly efficacious in management of oxidative stress and its sequelae.

Oxidative Damage to Sperm DNA: Clinical Implications

Background: Sperm DNA is susceptible to oxidative damage due to intrinsic and extrinsic factors which cause oxidative stress and due to limited DNA damage detection and repair mechanism. Reactive oxygen species (ROS) are the chief cause of sperm DNA damage. So, this study was planned to assess oxidative stress levels and correlate with sperm DNA damage. Material and Method: The study included 35 men who had fathered a child in the last one year and 54 male partners of couple experiencing primary infertility. Semen analysis was done according to World Health Organization (1999) criteria. ROS measurement was done by direct chemiluminescence method using luminol as a probe. DNA damage was assessed by sperm chromatin structure assay (SCSA) and expressed as percentage DFI. 8- Hydroxy-2’-deoxyguanosine (8-OHdG) estimation was carried out by competitive ELISA. Results: The seminal ROS level (RLU/sec/million sperm) was significantly higher (40.52 ± 18.32) in infertile men as compared to fertile controls (14.04 ± 6.67) (p<0.0001). The mean values of 8-OHdG levels (pg/ml) were also significantly higher in patients (30.92 ± 3.27) as compared to fertile controls (14.29 ± 2.24) (p<0.0001) and mean DFI (%) of infertile men was found to be 35.48 ± 12.95, which was higher as compared to controls (24.18 ± 8.76). There was a strong positive correlation between these parameters. Conclusion: Majority of the sperm DNA damage in infertile men is caused by oxidative damage to the genomic DNA. In presence of limited DNA damage detection and repair mechanism in sperm, prevention of oxidative stress by simple lifestyle interventions may actually be therapeutic.

Oxidative DNA damage and reduced expression of DNA repair genes: Role in primary open angle glaucoma (POAG)

ABSTRACT Background: Controversy exists regarding the role of oxidative DNA damage and DNA repair in primary open angle glaucoma (POAG). We performed a case control study to test the hypothesis that oxidative DNA damage and base excision repair (BER) genes PARP1 and OGG1 are involved in POAG pathogenesis. Materials and Methods: The study included 116 POAG patients and 116 cataract patients as controls. The 8-hydroxy-2’-deoxyguanosine (8-OHdG) levels were measured by ELISA. RNA was extracted from blood by Trizol and converted to cDNA. The relative quantification of PARP1 and OGG1 genes normalized to β-actin was calculated by the 2−ΔCt method. Comparisons between groups were done by student’s t-test and correlation between parameters was seen by Pearson correlation coefficient. All p values less than 0.05 were considered significant. Results: Mean levels of 8-OHdG were (patients v/s controls) 19.53 ± 1.40 vs. 15.0 ± 2.6 ng/ml in plasma and 8.55 ± 1.94 vs. 5.15 ± 1.09 ng/ml in aqueous humor (p < 0.0001). Expression levels of PARP1 (0.44 ± 0.05 vs. 0.88 ± 0.04) and OGG1 (0.46 ± 0.05 vs. 0.8 ± 0.01) were significantly (p < 0.0001) less in the patients than controls. There was a significant negative correlation between the expression levels of PARP1 and OGG1 with plasma and aqueous 8-OHdG. There was a strong positive correlation between plasma and aqueous 8-OHdG levels. Conclusion: These results support our hypothesis that oxidative stress-induced DNA damage is associated with POAG. Increased oxidative DNA damage in POAG may be attributed to decreased expression of DNA repair enzymes of the BER pathway.

Neurodegenerative Eye Disorders: Role of Mitochondrial Dynamics and Genomics.

AbstractAs a major source of cellular energy, mitochondria are critical for optimal ocular function. They are also essential for cell differentiation and survival. Mitochondrial mutations and oxidative damage to the mitochondrial DNA are important factors underlying the pathology of many ocular disorders. With increasing age, mitochondrial DNA damage accumulates and results in several eye diseases. It is evident that the mitochondrial genome is more susceptible to stress and damage than the nuclear genome, as it lacks histone protection, a nucleotide excision repair system, and recombination repair, and it is the source and target of free radicals. Accumulation of mitochondrial mutations beyond a certain threshold explains the marked variations in phenotypes seen in mitochondrial diseases and the molecular mechanisms related to the pathogenesis of several chronic disorders in the eye. This review details the structure and function of mitochondria and the mitochondrial genome along with the mitochondrial involvement in various neurodegenerative ophthalmic disorders.

Genetic, Biochemical and Clinical Insights into Primary Congenital Glaucoma

ABSTRACT Glaucoma is an irreversible form of optic neuropathy in which the optic nerve suffers damage in a characteristic manner with optic nerve cupping and retinal ganglion cell death. Primary congenital glaucoma (PCG) is an idiopathic irreversible childhood blinding disorder which manifests at birth or within the first year of life. PCG presents with a classical triad of symptoms (viz epiphora, photophobia and blepharospasm) though there are many additional symptoms, including large eye ball and hazy cornea. The only anatomical anomaly found in PCG is trabecular meshwork (TM) dysgenesis. PCG is an inheritable disease with established genetic etiology. It transmits through autosomal recessive mode. A number of cases are sporadic also. Mutations in many genes have been found to be causative in PCG and many are yet to be found. Mutations in cytochrome P4501B1 (CYP1B1) gene have been found to be the predominant cause of PCG. Other genes that have been implicated in PCG etiology are myocilin, Forkhead-related transcription factor C1 (FOXC1) and latent transforming growth factor beta-binding protein 2 (LTBP2). Mutations in these genes have been reported from many parts of the world. In addition to this, mitochondrial genome mutations are also thought to be involved in its pathogenesis. There appears to be some mechanism involving more than one genetic factor. In this review, we will discuss the various clinical, biochemical and genetic aspects of PCG. We emphasize that etiology of PCG does not lie in a single gene or genetic factor. Research needs to be oriented into a direction where gene-gene interactions, ocular embryology, ophthalmic metabolism and systemic oxidative status need to be studied in order to understand this disorder. We also accentuate the need for ophthalmic genetic facilities in all ophthalmology setups. How to cite this article: Faiq M, Sharma R, Dada R, Mohanty K, Saluja D, Dada T. Genetic, Biochemical and Clinical Insights into Primary Congenital Glaucoma. J Current Glau Prac 2013;7(2):66-84.

Molecular Diagnostics and Genetic Counseling in Primary Congenital Glaucoma

ABSTRACT Primary congenital glaucoma (PCG) is a childhood irreversible blinding disorder with onset at birth or in the first year of life. It is characterized by the classical traid of symptoms viz. epiphora (excessive tearing), photophobia (hypersensitivity to light) and blepharospasm (inflammation of eyelids). The only anatomical defect seen in PCG is trabecular meshwork dysgenesis. PCG shows autosomal recessive mode of inheritance with considerable number of sporadic cases. The etiology of this disease has not been fully understood but some genes like CYP1B1, MYOC, FOXC1, LTBP2 have been implicated. Various chromosomal aberrations and mutations in mitochondrial genome have also been reported. Molecular biology has developed novel techniques in order to do genetic and biochemical characterization of many genetic disorders including PCG. Techniques like polymerase chain reaction, single strand conformational polymorphism and sequencing are already in use for diagnosis of PCG and other techniques like protein truncation testing and functional genomics are beginning to find their way into molecular workout of this disorder. In the light of its genetic etiology, it is important to develop methods for genetic counseling for the patients and their families so as to bring down its incidence. In this review, we ought to develop a genetic insight into PCG with possible use of molecular biology and functional genomics in understanding the disease etiology, pathogenesis, pathology and mechanism of inheritance. We will also discuss the possibilities and use of genetic counseling in this disease. How to cite this article: Faiq M, Mohanty K, Dada R, Dada T. Molecular Diagnostics and Genetic Counseling in Primary Congenital Glaucoma. J Current Glau Prac 2013;7(1):25-35.