Guidelines in the era of realistic medicine

No abstract available

A Rapid, Simple DNA Mismatch Repair Substrate Construction Method

A more flexible and higher-yielding in vitro DNA mismatch repair (MMR) substrate construction method, which was developed initially by Wang and Hays, is described for the construction of a nucleotide-based chemical mismatch (G/IU) and a G/T mismatch. Our modifications use the combination of two endonuclease enzymes (NheI and BciVI) and two new redesigned plasmids (pWDAH1A and pWDSH1B). In our modified methodology, plasmids are initially digested with the nicking endonucleases, followed by the streptavidin treatment. The mismatch-containing oligo is then annealed to the gap DNA and finally ligated to produce a mismatch-containing DNA substrate. We report a high efficiency (up to 90%) of these mismatch substrates and confirm recognition using a functional assay. These modifications, coupled with the use of the redesigned plasmids, can be applied for the construction of other types of chemically induced mismatches as well as insertion-deletion loops for future in vitro studies of MMR processing by our group and others

Covering the Plague Years: Four Approaches to the AIDS Beat

AIDS reporting has changed dramatically since 1981. But it was not until mid-1985, when Rock Hudson was diagnosed with the disease, that media outlets began playing the epidemic as a story of major proportions. Because almost no major media institution embraced the AIDS story as an important issue, coverage of the epidemic was often the result of a reporter\u27s initiative. Consequently, the connection the individual journalist had with the epidemic became a much stronger influence on what appeared in the news and on what Americans knew about the crisis than in any other recent major health story. This article examines how four prominent journalists covered the disease. The reporting by the San Francisco Chronicle\u27s Randy Shilts, a gay man aligned with a political faction in the city \u27s homosexual community, reflected that affiliation. Jim Bunn, a heterosexual reporterfor KPIX-TV in San Francisco, brought to the epidemic the fear that it would spread to the larger, heterosexual population, and worked hard to get the word out about that possibility. The New York Times\u27s Dr. Lawrence Altman viewed the epidemic from his perspective as a traditional medical doctor — maintaining a professional distance from the tragedy. And National Public Radio\u27s Laurie Garrett, a scientist as well as a heterosexual woman politically in touch with the gay community, took a compassionate, informed stance. Almost all the coverage of these journalists had discernible policy impacts

Ontological Dumpster Diving

Throughout the literature on personal identity, the term ‘fourdimensionalism’ is poorly understood. Indeed, Mark Johnston deploys the concept of ontological trash to show that there is no feasible four-dimensionalist account of a person as an object entirely within spacetime, but he does not consider how any particular theory of spacetime or four-dimensionalism comes to bear on personhood. In this paper I will explain this line of reasoning, clarify fourdimensionalism, and show that there is a feasible account of personhood available on four-dimensionalism. In the introduction, I explain the concept of ontological trash and its threat to personhood. In the first section, I explain the concept of time dilation and use it, in conjunction with ontological trash, to prove that a person’s life does not have an unqualiοed temporal duration. In the second section, I summarise Cody Gilmore’s analysis of four-dimensionalism and explain how it comes to bear on persistence. In the third section, I sketch a new way to escape ontological trash in light of four-dimensionalism. In the fourth section, I apply this response to personhood, arguing that persons exist fully within spacetime and can withstand almost any psychological change. In the conclusion, I reflect on avenues for future research

Impact of Autophagy on Chemotherapy and Radiotherapy Mediated Tumor Cytotoxicity: “To Live or not to Live”

Autophagy, a highly regulated cell “self-eating” pathway, is controlled by the action of over 34 autophagy-related proteins (collectively termed Atgs). Although they are fundamentally different processes, autophagy and apoptosis (type I programmed cell death), under certain circumstances, can be regulated by common signaling mediators. Current cancer therapies including chemotherapy and ionizing radiation are known to induce autophagy within tumor cells. However, autophagy plays a dual role of either pro-cell survival or pro-cell death in response to these cancer treatments, depending on the cellular context and the nature of the treatment. We review the current basic and translational cancer research literature on how autophagy impacts tumor cell survival (“to live”) and death (“not to live”) following treatment as well as the role of chemical mediators of autophagy

In silico and in vitro screening for potential anticancer candidates targeting GPR120

The G-protein coupled receptor - GPR120 has recently been implicated as a novel target for colorectal cancer (CRC) and other cancer managements. In this study, a homology model of GPR120S (short isoform) was generated to identify potential anti-cancer compounds targeting the GPR120 receptor using a combined in silico docking-based virtual screening (DBVS), structure-activity relationships (SAR) and in vitro screening approach. SPECS database of synthetic chemical compounds (~350,000) was screened using the developed GPR120S model to identify molecules binding to the orthosteric binding pocket followed by an AutoDock SMINA rigid-flexible docking protocol. The best 13 hit molecules were then tested in vitro to evaluate their cytotoxic activity against SW480 - human CRC cell line expressing GPR120. The test compound 1 (3-(4-methylphenyl)-2-[(2-oxo-2-phenylethyl)sulfanyl]-5,6-dihydrospiro(benzo[h]quinazoline-5,1\u27-cyclopentane)-4(3H)-one) showed ~ 90% inhibitory effects on cell growth with micromolar affinities (IC50 = 23.21-26.69 µM). Finally, SAR analysis of compound 1 led to the identification of a more active compound from the SPECS database showing better efficacy during cell-based cytotoxicity assay -5 (IC50 = 5.89-6.715 µM), while a significant reduction in cytotoxic effects of 5 was observed in GPR120-siRNA pre-treated SW480 cells. The GPR120S homology model generated, and SAR analysis conducted by this work discovered a potential chemical scaffold, dihydrospiro(benzo[h]quinazoline-5,1\u27-cyclopentane)-4(3H)-one, which will aid future research on anti-cancer drug development for CRC management

Structure based prediction of a novel GPR120 antagonist based on pharmacophore screening and molecular dynamics simulations

The G-protein coupled receptor, GPR120, has ubiquitous expression and multifaceted roles in modulating metabolic and anti-inflammatory processes. Recent implications of its role in cancer progression have presented GPR120 as an attractive oncogenic drug target. GPR120 gene knockdown in breast cancer studies revealed a role of GPR120-induced chemoresistance in epirubicin and cisplatin-induced DNA damage in tumour cells. Higher expression and activation levels of GPR120 is also reported to promote tumour angiogenesis and cell migration in colorectal cancer. Some agonists targeting GPR120 have been reported, such as TUG891 and Compound39, but to date development of small-molecule inhibitors of GPR120 is limited. Herein, following homology modelling of the receptor a pharmacophore hypothesis was derived from 300 ns all-atomic molecular dynamics (MD) simulations on apo, TUG891-bound and Compound39-bound GPR120S (short isoform) receptor models embedded in a water solvated lipid bilayer system. We performed comparative MD analysis on protein–ligand interactions between the two agonist and apo simulations on the stability of the “ionic lock” – a Class A GPCRs characteristic of receptor activation and inactivation. The detailed analysis predicted that ligand interactions with W277 and N313 are critical to conserve the “ionic-lock” conformation (R136 of Helix 3) and prevent GPR120S receptor activation. The results led to generation of a W277 and N313 focused pharmacophore hypothesis and the screening of the ZINC15 database using ZINCPharmer through the structure-based pharmacophore. 100 ns all-atomic molecular dynamics (MD) simulations were performed on 9 small molecules identified and Cpd 9, (2-hydroxy-N-{4-[(6-hydroxy-2-methylpyrimidin-4-yl) amino] phenyl} benzamide) was predicted to be a small-molecule GPR120S antagonist. The conformational results from the collective all-atomic MD analysis provided structural information for further identification and optimisation of novel druggable inhibitors of GPR120S using this rational design approach, which could have future potential for anti-cancer drug development studies

Analyses and comparison of counter-movement jump performance and self-rated recovery in state under-18s Australian Rules Football players during a national championship

Recovery of team sport athletes during multiple competitive games is an important area for strength and conditioning coaches to monitor as it facilitates for athletes to be ready to perform (11,13). Utilising athletic performance data in conjunction with self-rated reporting measures can help determine if in fact a player or team has recovered sufficiently or shown a trend towards recovery prior to a competitive match (11). Positive improvement in recovery variables can provide confidence in the effectiveness of recovery methods used and assist in determining the training schedule in order to positively manipulate the fitness-fatigue relationship (3). Various methods of analysing the recovery of athletes have been reported in the literature and are available to the strength and conditioning coach. These include subjective, self-rated scales and perceived level of recovery questionnaires (11,12,13). Athletic performance measures during exercises such as the counter movement jump (CMJ) have also been analysed, predominantly utilising force plates to obtain kinetic data. (5,13,14). However, such equipment can be difficult to transport, requires continual calibration and is costly to purchase. A linear transducer can provide important information on CMJ variables in the assessment of athletic movements and due to its size and portability could serve as a valuable tool to assist strength and conditioning coaches, (8,10), and potentially enable the monitoring of recovery. Previous studies have investigated the fatigue effects of competitive games in various sports (11,13,14) including Australian Rules Football (AFL) at the senior elite league level (5, 6). To the authors&rsquo; knowledge, however, there is yet to be a study investigating the recovery response in AFL players, specifically in players 18 years and under competing in the National Under 18s Championships. Australian Rules football is an extremely physically demanding and fatiguing sport where players participate in games time exceeding 120 minutes duration, covering large distances (~12-18km, position dependent) with many high intensity efforts performed at random times throughout the game (2,6,16). Hence, it would seem pertinent to analyse the fatigue effects of competitive matches in an Australian Rules Under-18&rsquo;s National Championship and the subsequent recovery from these games. The aim of this study was to analyse and compare two self-rated subjective measures of recovery; they being muscle soreness (MS) of the lower body, overall perceived total recovery (TR), and the performance measure of peak velocity (PV) obtained from a CMJ analysed with a linear transducer. Data collection occurred between rounds four and five of the Australian Football League Under-18&rsquo;s National Championship, representing a four-day recovery analysis period between matches.<br /