Obstructive Sleep Apnea and Ophthalmic Disorders-Clinical Implications.

Sleep is essential for physical, mental and emotional well being. Body systems require sleep of good quality and quantity for their proper functioning. There are several sleep disorders. Obstructive sleep apnea hypopnea syndrome (OSAHS) is one of the most important disorders identified in the last 50 years. The disorder has systemic ill effects by virtue of cyclical hypoxia and sympathetic stimulation. It is a risk factor for the development of hypertension, ischemic heart disease, type 2 diabetes mellitus, stroke and dementia. Retina being the highest oxygen consuming part of the body, is particularly vulnerable to the effects of hypoxia. Several eye disorders have been identified to be associated with OSAHS. In clinical practice Identifying and treating sleep disorders have been rewarding.

Transcriptional regulation of the human DNA Methyltransferase (dnmt1) gene.

DNA methylation is an important component of the epigenetic control of genome functions. Understanding the regulation of the DNA Methyltransferase (dnmt1) gene expression is critical for comprehending how DNA methylation is coordinated with other critical biological processes. In this paper, we investigate the transcriptional regulatory region of the human dnmt1 gene using a combination of RACE, RNase protection analysis and CAT assays. We identified one major and three minor transcription initiation sites in vivo (P1-P4), which are regulated by independent enhancers and promoter sequences. The minimal promoter elements of P1, P2 and P4 are mapped within 256 bp upstream of their respective transcription initiation sites. P1 is nested within a CG-rich area, similar to other housekeeping genes, whereas P2-P4 are found in CG-poor areas. Three c-Jun-dependent enhancers are located downstream to P1 and upstream to P2-P4, thus providing a molecular explanation for the responsiveness of dnmt1 to oncogenic signals that are mediated by the Ras-c-Jun oncogenic signaling pathway.

Effect of metanil yellow, orange II and their blend on hepatic xenobiotic metabolizing enzymes in rats.

The effects of Metanil yellow, Orange II and their blend on hepatic xenobiotic metabolizing enzymes were compared. Parenteral administration of Metanil yellow and Orange II to rats at a dose of 80 mg/kg body weight for 3 days caused a significant induction of ethoxyresorufin-O-deethylase (40-190%), aniline hydroxylase (27-92%), aryl hydrocarbon hydroxylase (50-62%) and aminopyrine N-demethylase (42-49%) activities. Metanil yellow and Orange II brought about a substantial increase in cytosolic quinone reductase (34-82%) and glutathione S-transferase (23-43%) activities and significant depletion of glutathione levels with a concomitant increase in lipid peroxide formation. A blend (1:1) of Metanil yellow and Orange II showed a synergistic or additive effect on these hepatic parameters, suggesting that the addition of these two prohibited dyes together in foodstuffs may give rise to more toxic effects than are produced by each dye individually.

Metanil yellow: a bifunctional inducer of hepatic phase I and phase II xenobiotic-metabolizing enzymes.

Metanil yellow, a non-permitted food colour, has been found in various foodstuffs. The induction potential of metanil yellow on hepatic microsomal cytochrome P-450 (P-450)-dependent monooxygenases and cytosolic detoxification enzymes, namely, glutathione S-transferase (GST) and quinone reductase (QR), was investigated. Oral administration of metanil yellow (430 mg/kg body weight) to four animals for seven days caused significant induction of hepatic P-450 (48%) and its dependent aryl hydrocarbon hydroxylase (100%) activity and cytosolic GST (136%) and QR (92%) activities. Parenteral administration of metanil yellow (80 mg/kg body weight) to another set of four animals for 3 days resulted in higher induction of ethoxyresorufin-O-deethylase (228%) as compared to other monooxygenases (64-92%), while GST and QR were also found to be induced (59-95%). Spectra of metanil yellow-induced microsomes showed an increase in P-450 with a shift of 2.2 nm in the soret region. The results suggest that metanil yellow acts as a bifunctional inducer of specific isozymes of P-450 and cytosolic enzymes and thus may involve the cytosolic aryl hydrocarbon (Ah) receptor for this type of induction.

Pulmonary arteriovenous malformations.

Pulmonary arteriovenous malformations rarely present in childhood. Two cases are presented in this report. Both the cases presented clinically with cyanosis and clubbing without a cardiac murmur. The second case had cerebral abscess in addition. Both the cases underwent a contrast-enhanced echocardiography which suggested the presence of pulmonary arteriovenous malformations. The first case also underwent 99mTc radionuclide scan and pulmonary angiography. The cases are being reported for their characteristic clinical features and for emphasizing the role of non-invasive modalities like contrast-enhanced echocardiography and radionuclide scan in reaching the diagnosis.

Perioperative management of laryngotracheobronchial injury: our experience in a level 1 trauma centre.

PURPOSE: Laryngotracheobronchial injuries (LTBI) are serious injuries because of their consequences in terms of ventilation, coupled with the severity of other injuries associated with them. We share our experience in managing these patients perioperatively in our level 1 trauma centre. METHODS: A retrospective analysis of the records of 30 patients with LTBI who presented at Jai Prakash Narayan Apex Trauma Center (JPNATC) from December 2007 to February 2011 was done. The demographics, mechanism of injury, clinical presentation, diagnostic modalities, anaesthetic management and outcome in these patients were reviewed. RESULTS: Intrathoracic location of the injury and Injury Severity Score (ISS) had a direct correlation with the outcome of the patients. The overall mortality was 6.7 %. CONCLUSION: Meticulous examination, details about the mechanism of injury, careful diagnostic evaluation, and skilful airway and surgical management are necessary for a better outcome in patients with airway injuries. A high degree of suspicion in occult injuries and liberal use of a fibreoptic bronchoscope aids diagnosis and management. Prompt airway management in the pre-hospital setting before transfer to a higher level trauma centre ensures better outcomes.

Genomic structure of the human DNA methyltransferase gene.

We determined the genomic structure of the gene encoding human DNA methyltransferase (DNA MTase). Six overlapping human genomic DNA clones which include all of the known cDNA sequence were isolated. Analysis of these clones demonstrates that the human DNA MTase gene consists of at least 40 exons and 39 introns spanning a distance of 60 kilobases. Elucidation of the chromosomal organization of the human DNA MTase gene provides the template for future structure-function analysis of the properties of mammalian DNA MTase.

A conserved 3'-untranslated element mediates growth regulation of DNA methyltransferase 1 and inhibits its transforming activity.

Ectopic expression of DNA methyltransferase 1 (DNMT1) has been proposed to play an important role in cancer. dnmt1 mRNA is undetectable in growth-arrested cells but is induced upon entrance into the S phase of the cell cycle, and until now, the mechanisms responsible for this regulation were unknown. In this report, we demonstrate that the 3'-untranslated region (3'-UTR) of the dnmt1 mRNA can confer a growth-dependent regulation on its own message as well as a heterologous beta-globin mRNA. Our results indicate that a 54-nucleotide highly conserved element within the 3'-UTR is necessary and sufficient to mediate this regulation. Cell-free mRNA decay experiments demonstrate that this element increases mRNA turnover rates and does so to a greater extent in the presence of extracts prepared from arrested cells. A specific RNA-protein complex is formed with the 3'-UTR only in growth-arrested cells, and a UV cross-linking analysis revealed a 40-kDa protein (p40), the binding of which is dramatically increased in growth-arrested cells and is inversely correlated with dnmt1 mRNA levels as cells are induced into the cell cycle. Although ectopic expression of human DNMT1 lacking the 3'-UTR can transform NIH-3T3 cells, inclusion of the 3'-UTR prevents transformation. These results support the hypothesis that deregulated expression of DNMT1 with the cell cycle is important for cellular transformation.

Effect of oral and parenteral administration of metanil yellow on some hepatic and intestinal biochemical parameters.

Metanil yellow, a non-permitted colour for food commodities, is used in the leather, paper and textile industries. In this paper the effect of oral and parenteral administration of Metanil yellow on hepatic and intestinal biochemical parameters was investigated. Oral administration of Metanil yellow (430 mg kg-1 body wt.) for 7 days caused significant depletion of hepatic and intestinal glutathione levels (33-52%) with a concomitant increase in lipid peroxidation (49-121%). Metanil yellow treatment for 7 days also led to a significant increase in cytochrome P-450 (P-450)-dependent aryl hydrocarbon hydroxylase (AHH) activity (99-223%) in the liver and intestine. Cytosolic glutathione-S-transferase (GST) (32-136%) and quinone reductase (QR) (20-92%) activities were also found to be substantially induced in hepatic and intestinal tissues following oral treatment of Metanil yellow. It is interesting to note that oral treatment of Metanil yellow showed a greater response in cytosolic enzymes of hepatic tissue as compared to intestine. Single parenteral administration of Metanil yellow (80 mg kg-1 body wt.) caused significant induction of P-450 and its dependent monooxygenases. Even after 5 days of single parenteral administration of Metanil yellow, hepatic AHH activity showed an elevation of 48% while other monooxygenases were marginally increased. Cytosolic GST and QR showed respective peak inductions of 92% and 60% after 2 and 3 days of parenteral administration of Metanil yellow, which levels off after the 5th day. It can be concluded that Metanil yellow acts as an inducer of a specific form of microsomal P-450 and cytosolic GST and QR, which may involve a cytosolic Ah receptor.

Deciphering the language of the genome.

The non-coding DNA in eukaryotic genomes encodes a language which programs organismal growth and development. We show that a linguistic and cryptographic approach can be used to deduce the syntax of this programming language for gene regulation and to compile a dictionary of enhancers which form its words.

A mammalian protein with specific demethylase activity for mCpG DNA.

DNA-methylation patterns are important for regulating genome functions, and are determined by the enzymatic processes of methylation and demethylation. The demethylating enzyme has now been identified: a mammalian complementary DNA encodes a methyl-CpG-binding domain, bears a demethylase activity that transforms methylated cytosine bases to cytosine, and demethylates a plasmid when the cDNA is translated or transiently transfected into human embryonal kidney cells in vitro. The discovery of this DNA demethylase should provide a basis for the molecular and developmental analysis of the role of DNA methylation and demethylation.

DNA methylation is a reversible biological signal.

The pattern of DNA methylation plays an important role in regulating different genome functions. To test the hypothesis that DNA methylation is a reversible biochemical process, we purified a DNA demethylase from human cells that catalyzes the cleavage of a methyl residue from 5-methyl cytosine and its release as methanol. We show that similar to DNA methyltransferase, DNA demethylase shows CpG dinucleotide specificity, can demethylate mdCpdG sites in different sequence contexts, and demethylates both fully methylated and hemimethylated DNA. Thus, contrary to the commonly accepted model, DNA methylation is a reversible signal, similar to other physiological biochemical modifications.

Inhibition of tumorigenesis by a cytosine-DNA, methyltransferase, antisense oligodeoxynucleotide.

This paper tests the hypothesis that cytosine DNA methyltransferase (DNA MeTase) is a candidate target for anticancer therapy. Several observations have suggested recently that hyperactivation of DNA MeTase plays a critical role in initiation and progression of cancer and that its up-regulation is a component of the Ras oncogenic signaling pathway. We show that a phosphorothioate-modified, antisense oligodeoxynucleotide directed against the DNA MeTase mRNA reduces the level of DNA MeTase mRNA, inhibits DNA MeTase activity, and inhibits anchorage independent growth of Y1 adrenocortical carcinoma cells ex vivo in a dose-dependent manner. Injection of DNA MeTase antisense oligodeoxynucleotides i.p. inhibits the growth of Y1 tumors in syngeneic LAF1 mice, reduces the level of DNA MeTase, and induces demethylation of the adrenocortical-specific gene C21 and its expression in tumors in vivo. These results support the hypothesis that an increase in DNA MeTase activity is critical for tumorigenesis and is reversible by pharmacological inhibition of DNA MeTase.

Identification of initiation sites for DNA replication in the human dnmt1 (DNA-methyltransferase) locus.

Vertebrates have developed multiple mechanisms to coordinate the replication of epigenetic and genetic information. Dnmt1 encodes the maintenance enzyme DNA-methyltransferase, which is responsible for propagating the DNA methylation pattern and the epigenetic information that it encodes during replication. Direct sequence analysis and bisulfite mapping of the 5' region of DNA-methyltransferase 1 (dnmt1) have indicated the presence of many sequence elements associated with previously characterized origins of DNA replication. This study tests the hypothesis that the dnmt1 region containing these elements is an origin of replication in human cells. First, we demonstrate that a vector containing this dnmt1 sequence is able to support autonomous replication when transfected into HeLa cells. Second, using a gel retardation assay, we show that it contains a site for binding of origin-rich sequences binding activity, a recently purified replication protein. Finally, using competitive polymerase chain reaction, we show that replication initiates in this region in vivo. Based on these lines of evidence, we propose that initiation sites for DNA replication are located between the first intron and exon 7 of the human dnmt1 locus.