RNA Interference Technology — Applications and Limitations

RNA interference (RNAi), an evolutionarily conserved mechanism triggered by double-stranded RNA (dsRNA), causes gene silencing in a sequence-specific manner. RNAi evolved naturally to mediate protection from both endogenous and exogenous pathogenic nucleic acids and to modulate gene expression. Multiple technological advancements and precision in gene targeting have allowed a plethora of potential applications, ranging from targeting infections in crop plants to improving health in human patients, which have been reviewed in this chapter

Metabolic Rewiring by Loss of Sirt5 Promotes Kras-induced Pancreatic Cancer Progression.

BACKGROUND & AIMS: SIRT5 plays pleiotropic roles via post-translational modifications, serving as a tumor suppressor, or an oncogene, in different tumors. However, the role SIRT5 plays in the initiation and progression of pancreatic ductal adenocarcinoma (PDAC) remains unknown. METHODS: Published datasets and tissue arrays with SIRT5 staining were used to investigate the clinical relevance of SIRT5 in PDAC. Furthermore, to define the role of SIRT5 in the carcinogenesis of PDAC, we generated autochthonous mouse models with conditional Sirt5 knockout. Moreover, to examine the mechanistic role of SIRT5 in PDAC carcinogenesis, SIRT5 was knocked down in PDAC cell lines and organoids, followed by metabolomics and proteomics studies. A novel SIRT5 activator was utilized for therapeutic studies in organoids and patient-derived xenografts. RESULTS: SIRT5 expression negatively regulated tumor cell proliferation and correlated with a favorable prognosis in PDAC patients. Genetic ablation of Sirt5 in PDAC mouse models promoted acinar-to-ductal metaplasia, precursor lesions, and pancreatic tumorigenesis, resulting in poor survival. Mechanistically, SIRT5 loss enhanced glutamine and glutathione metabolism via acetylation-mediated activation of GOT1. A selective SIRT5 activator, MC3138, phenocopied the effects of SIRT5 overexpression and exhibited anti-tumor effects on human PDAC cells. MC3138 also diminished nucleotide pools, sensitizing human PDAC cell lines, organoids, and patient-derived xenografts to gemcitabine. CONCLUSIONS: Collectively, we identify SIRT5 as a key tumor suppressor in PDAC, whose loss promotes tumorigenesis through increased non-canonical utilization of glutamine via GOT1, and that SIRT5 activation is a novel therapeutic strategy to target PDAC

Short-term effect of multifidus muscle intramuscular electrical stimulation in chronic low back pain

Interventions for mechanical low back pain (LBP) include physical therapy and pharmaceutical management, as well as surgical intervention. Although the paraspinal muscles have been identified as important spinal stabilizers and are targeted during physical therapy, diminished activation of this muscle can lead to biomechanical dysfunction and lead to mechanical LBP. Intramuscular electrical stimulation (IMES) with low-frequency (2 Hz) current facilitates release of endogenous opioids which promotes analgesia and causes activation of multifidus muscle and improves the changes produced by muscular dysfunction. After single session of intervention, the patient reported that her pain levels were 0 and 1 on a numeric pain scale of 0-10 during rest and walking, respectively. The average score on the Oswestry LBP Disability Questionnaire was reduced from 43/50 to 11/50. Thus, IMES can be used as an effective therapeutic strategy to directly target the deep multifidus muscle and to treat mechanical LBP produced due to multifidus dysfunction

Core Strength as a Paramount Contributor for Potential Upper Limb Isometric Strength - A Correlational Study

It is believed that a strong core will enable an athlete to effectively transfer forces from the lower extremities through the torso to the upper extremities. Control of the shoulder girdle force is important for the proper function of the upper extremities, although stabilising forces from the trunk and pelvis are also important. The purpose of this study is to determine the association between shoulder isometric push-up/pull-down strength and isometric strength of the abdominal muscles. Using the HUR Rehab-Line 5310 and 5120, isometric upper limb and core strength was measured. The statistical analysis revealed a moderate but significant positive association between the strength of the back muscles and isometric push-ups. Additionally, there was a weak and significant connection between isometric push-ups and abdominal strength. The study therefore draws the conclusion that there is a link between isometric upper limb performance and core musculature strength/stability. The abdominal and back muscles' isometric strength is closely tied to the isometric push-up, which is attributable to the contraction of the Pectoralis Major, Deltoid, and Triceps Brachii. The only muscle group identified to have a substantial and positive correlation with isometric pull-down strength is the abdominal muscles

Study of various Bug Tracking Tools

As per the size of software increases in modern software organizations, number of defects is also increasing. Bugs are inevitable in software. Bugs reduce the reliability of software by increasing the risk. It is not always easy to track and mitigate issues caused by bugs. So as Bug tracking softwares are introduced. Bug tracking tools are a preventive measure that not only finds bugs but also do the debugging process. It handles cross communication between different team members and keeps the track of bug handling which makes it scalable. For a software industry it is very important to know about usage of any bug tracking software as per their requirements. This paper focuses on some most popular bug tracking tools that are currently being used in software firms. Presented study helps to select the tool that provides the best solution in context to the requirement of industry. A Comparative matrix is given to provide better analysis and visualization of same

Successful management of pulpo-periodontal lesion in maxillary lateral incisor with palatogingival groove using CBCT scan

Palatogingival groove is a rare developmental anomaly involving the lingual surface of the maxillary incisor and resulting in severe endodontic and periodontal lesions. This case report describes a multidisciplinary approach for the combined management of the endodontic and periodontal problems for successful rehabilitation of the involved tooth. Cone-beam computed tomography (CBCT) helped in correct diagnosis of the lesion and hence enabled effective treatment

Endodontic management of open apex using Biodentine as a novel apical matrix

Aim: Endodontic management of open apex using Biodentine as an apical matrix. Summary : An immature tooth with pulpal necrosis and periapical pathology imposes a great difficulty to the endodontist. Endodontic treatment options for such teeth consist of conventional apexification procedure with and without apical barriers. Biodentine™ is new calcium silicate based cement that exhibits physical and chemical properties similar to those described for certain Portland cement derivatives. This article demonstrates the use of the newer material, Biodentine as an apical matrix barrier in root end apexification procedure. This case reports present apexification and successful healing with the use of Biodentine as an apical barrier matrix. Conclusion : Apexification in one step using an apical plug of Biodentine can be considered a predictable treatment and may be an alternative to mineral trioxide aggregate apexification

Small-Scale Screening to Large-Scale Over-Expression of Human Membrane Proteins for Structural Studies

Membrane protein structural studies are frequently hampered by poor expression. The low natural abundance of these proteins implies a need for utilizing different heterologous expression systems. E. coli and yeast are commonly used expression systems due to rapid cell growth at high cell density, economical production, and ease of manipulation. Here we report a simplified, systematically developed robust strategy from small-scale screening to large-scale over-expression of human integral membrane proteins in the mammalian expression system for structural studies. This methodology streamlines small-scale screening of several different constructs utilizing fluorescence size-exclusion chromatography (FSEC) towards optimization of buffer, additives, and detergents for achieving stability and homogeneity. This is followed by the generation of stable clonal cell lines expressing desired constructs, and lastly large-scale expression for crystallization. These techniques are designed to rapidly advance the structural studies of eukaryotic integral membrane proteins including that of human membrane proteins