Transcriptome analysis using next generation sequencing reveals molecular signatures of diabetic retinopathy and efficacy of candidate drugs

Purpose: To define gene expression changes associated with diabetic retinopathy in a mouse model using next generation sequencing, and to utilize transcriptome signatures to assess molecular pathways by which pharmacological agents inhibit diabetic retinopathy. Methods: We applied a high throughput RNA sequencing (RNA-seq) strategy using Illumina GAIIx to characterize the entire retinal transcriptome from nondiabetic and from streptozotocin-treated mice 32 weeks after induction of diabetes. Some of the diabetic mice were treated with inhibitors of receptor for advanced glycation endproducts (RAGE) and p38 mitogen activated protein (MAP) kinase, which have previously been shown to inhibit diabetic retinopathy in rodent models. The transcripts and alternatively spliced variants were determined in all experimental groups. Results: Next generation sequencing-based RNA-seq profiles provided comprehensive signatures of transcripts that are altered in early stages of diabetic retinopathy. These transcripts encoded proteins involved in distinct yet physiologically relevant disease-associated pathways such as inflammation, microvasculature formation, apoptosis, glucose metabolism, Wnt signaling, xenobiotic metabolism, and photoreceptor biology. Significant upregulation of crystallin transcripts was observed in diabetic animals, and the diabetes-induced upregulation of these transcripts was inhibited in diabetic animals treated with inhibitors of either RAGE or p38 MAP kinase. These two therapies also showed dissimilar regulation of some subsets of transcripts that included alternatively spliced versions of arrestin, neutral sphingomyelinase activation associated factor (Nsmaf), SH3-domain GRB2-like interacting protein 1 (Sgip1), and axin. Conclusions: Diabetes alters many transcripts in the retina, and two therapies that inhibit the vascular pathology similarly inhibit a portion of these changes, pointing to possible molecular mechanisms for their beneficial effects. These therapies also changed the abundance of various alternatively spliced versions of signaling transcripts, suggesting a possible role of alternative splicing in disease etiology. Our studies clearly demonstrate RNA-seq as a comprehensive strategy for identifying disease-specific transcripts, and for determining comparative profiles of molecular changes mediated by candidate drugs. Diabetes has emerged as a major worldwide public health concern, and the number of diabetics is estimated to exceed 400 million by the year 203

Rd9 Is a Naturally Occurring Mouse Model of a Common Form of Retinitis Pigmentosa Caused by Mutations in RPGR-ORF15

Animal models of human disease are an invaluable component of studies aimed at understanding disease pathogenesis and therapeutic possibilities. Mutations in the gene encoding retinitis pigmentosa GTPase regulator (RPGR) are the most common cause of X-linked retinitis pigmentosa (XLRP) and are estimated to cause 20% of all retinal dystrophy cases. A majority of RPGR mutations are present in ORF15, the purine-rich terminal exon of the predominant splice-variant expressed in retina. Here we describe the genetic and phenotypic characterization of the retinal degeneration 9 (Rd9) strain of mice, a naturally occurring animal model of XLRP. Rd9 mice were found to carry a 32-base-pair duplication within ORF15 that causes a shift in the reading frame that introduces a premature-stop codon. Rpgr ORF15 transcripts, but not protein, were detected in retinas from Rd9/Y male mice that exhibited retinal pathology, including pigment loss and slowly progressing decrease in outer nuclear layer thickness. The levels of rhodopsin and transducin in rod outer segments were also decreased, and M-cone opsin appeared mislocalized within cone photoreceptors. In addition, electroretinogram (ERG) a- and b-wave amplitudes of both Rd9/Y male and Rd9/Rd9 female mice showed moderate gradual reduction that continued to 24 months of age. The presence of multiple retinal features that correlate with findings in individuals with XLRP identifies Rd9 as a valuable model for use in gaining insight into ORF15-associated disease progression and pathogenesis, as well as accelerating the development and testing of therapeutic strategies for this common form of retinal dystrophy

The spine in Paget’s disease

Paget’s disease (PD) is a chronic metabolically active bone disease, characterized by a disturbance in bone modelling and remodelling due to an increase in osteoblastic and osteoclastic activity. The vertebra is the second most commonly affected site. This article reviews the various spinal pathomechanisms and osseous dynamics involved in producing the varied imaging appearances and their clinical relevance. Advanced imaging of osseous, articular and bone marrow manifestations of PD in all the vertebral components are presented. Pagetic changes often result in clinical symptoms including back pain, spinal stenosis and neural dysfunction. Various pathological complications due to PD involvement result in these clinical symptoms. Recognition of the imaging manifestations of spinal PD and the potential complications that cause the clinical symptoms enables accurate assessment of patients prior to appropriate management

Relationship of subjective well-being with social support and meaning in life among college students

This study sought to determine the relationship and predictive influence of social support and meaning in life on the components of subjective well-being (life satisfaction, positive affect and negative affects) among the first year college students. The participants were 141 (81females and 60 males) first year college students from Metro Manila. The data collected from four standardized instruments, namely: Student Life Satisfaction Scale (SLSS) of Huebner, (1991) Positive and Negative Affective Schedule (PANAS) of Watson, Clark, & Tellegen, (1988) Social Support Appraisal Scale (SSA) of Vaux, Phillips, Holley, Thompson, Williams, and Stewart (1986) and Purpose in Life Test (PIL) of Crumbaugh and Maholick (1969) were used to measure the components of subjective well-being of the respondents. Correlational design and multiple regression analysis were employed in this study. The findings of the study show that there was a significant correlation between social support and the components of subjective well-being (life satisfaction and positive affect) and also between meaning in life and the components of subjective well-being (life satisfaction and positive affect). Social support and meaning in life were not significantly correlated and predicted with the negative affective component of subjective well-being. The results of multiple regression analysis showed that social support and meaning in life were predictive of subjective well-being (life satisfaction, positive affect). The research findings further revealed that in the life of first year college students, both interpersonal variable (social support) and intrapersonal variable (meaning in life) have influence on subjective well-being (life satisfaction and positive affect)

Prevalence and etiology for mandibular and mid-face bone fractures in a tertiary care hospital

Background: The rate of mandibular and mid-face bone fracture was found to vary among the population. This study sought to find out the prevalence of mandibular and mid-face bone fractures in a tertiary care hospital. Methods: A retrospective study was conducted over the period of three years. Subjects aged 5-80 years with mandibular and mid-face bone fractures admitted for surgical intervention were included. Type, pattern and major etiological factors for bone fracture were accounted for and subjected to statistical analysis. Results: Among the 103 cases, mandibular fracture was observed in 72 cases (45 single and 27 combined bones) with male dominance (69 males and 3 females; 23:1 ratio). The mid-face bone fractures were found in 41 cases within the male group (32 single and 9 combined bones). For the mandibular and mid-face bone fractures, the most prevalent age groups were the 19-29 and 30-49 years, respectively (p = 0.80). The parasymphysis/body (33%) and condyle/ramus (20%) were the bone fracture among the mandible observed with the highest frequency, while that of the mid-face bone was for the maxilla (43.7%) and zygoma (31.2%). Road traffic accidents (RTAs) were the major cause of these sorts of fractures in the 19-29 age group (62%; p = 0.001). Conclusion: The mandible was the single most common site of bone fracture because of RTAs in males 19-29 years of age. This emphasizes the need for public health awareness so that people follow general traffic rules and road safety measures. Prevalence was found to vary based on socioeconomic, cultural, and behavioral variations among the populations. [Arch Clin Exp Surg 2017; 6(4.000): 183-188

Intraoral Complications Associated with Multiple Myeloma: Case Reports

Plasma cell neoplasms such as multiple myeloma (MM), solitary plasmacytoma of the bone and extra medullar plasmacytoma are characterized by a monoclonal neoplastic proliferation. Oral manifestations in the form of oral and maxillofacial lesions are often the first sign of the disease. Because the symptoms are various, it is very difficult to diagnose MM in the oral and maxillofacial region. In this study, two male patients presenting with complaint of a painful oral lesion are reported [Med-Science 2015; 4(2.000): 2229-35

A Putative Leucine Zipper within the Herpes Simplex Virus Type 1 UL6 Protein Is Required for Portal Ring Formation▿

The herpes simplex virus type 1 UL6 protein forms a 12-subunit ring structure at a unique capsid vertex which functions as a conduit for encapsidation of the viral genome. To characterize UL6 protein domains that are involved in intersubunit interactions and interactions with other capsid proteins, we engineered a set of deletion mutants spanning the entire gene. Three deletion constructs, D-5 (Δ198-295), D-6 (Δ322-416), and D-LZ (Δ409-473, in which a putative leucine zipper was removed), were introduced into the viral genome. All three mutant viruses produced only B capsids, indicating a defect in encapsidation. Western blot analysis showed that the UL6 protein was present in the capsids isolated from two mutants, D-6 and D-LZ. The protein encoded by D-5, on the other hand, was not associated with capsids and was instead localized in the cytoplasm of the infected cells, indicating that this deletion affected the nuclear transport of the portal protein. The UL6 protein from the KOS strain (wild type) and the D-6 mutant were purified from insect cells infected with recombinant baculoviruses and shown to form ring structures as assessed by sucrose gradient centrifugation and electron microscopy. In contrast, the D-LZ mutant protein formed aggregates that sedimented throughout the sucrose gradient as a heterogeneous mixture and did not yield stable ring structures. A mutant (L429E L436E) in which two of the heptad leucines of the putative zipper were replaced with glutamate residues also failed to form stable rings. Our results suggest that the integrity of the leucine zipper region is important for oligomer interactions and stable ring formation, which in turn are required for genome encapsidation