Promotion of functional heterotrimeric type I collagen via transfection in osteogenesis imperfecta fibroblasts

Abstract only availableOsteogenesis imperfecta (OI) is a heritable disorder due to mutations in type I collagen. Normal type I collagen forms a heterotrimeric protein comprised of two pro1(I) chains and one pro2(I) chain [1(I)22(I)]. The osteogenesis imperfecta murine (oim) model mouse contains a single nucleotide deletion in the pro2(I) gene (COL1A2) resulting in non-functional pro2(I) chains and production of homotrimeric type I collagen containing three pro1(I) collagen chains, [1(I)3], resulting in small body size, increased bone fragility and altered bone mineralization. The overall goal of this study is to correct the oim defect by introducing normal COL1A2 genes into oim cells. Oim dermal fibroblasts were transfected with a series of COL1A2 gene constructs containing the full-length murine pro2(I) collagen cDNA driven by various lengths of the murine COL1A2 promoter (1.5kb, 3.0kb, and 6.0kb) along with a COL1A2 enhancer. These DNA constructs were cotransfected with pcDNA3 containing a neomycin resistance gene, which allows for selection of stably transfected cell lines. Various assays have been developed to monitor proa2(I) collagen expression at the DNA, RNA and protein levels. A PCR assay was used to confirm genomic incorporation of transgenic COL1A2 gene constructs and an RT-PCR assay used to confirm expression of normal pro2(I) collagen mRNA. Denaturing urea-SDS polyacrylamide gel electrophoresis along with Western blotting analyses using anti-murine 1(I) and 2(I) collagen antibodies were used to confirm normal pro2(I) collagen expression at the protein level as well as its incorporation into normal heterotrimeric type I collagen. All the necessary tools have been established for evaluating the efficacy of transfection. Currently, the first series of stably transfected oim cell lines are being expanded for analyses as described above. Although this study is aimed at 'fixing' the oim mutation via gene therapy, valuable data will also be collected regarding the effectiveness of the variable length promoter regions in enhancing the expression of the normal COL1A2 gene.Life Sciences Undergraduate Research Opportunity Progra

‘Fixed-axis’ magnetic orientation by an amphibian: non-shoreward-directed compass orientation, misdirected homing or positioning a magnetite-based map detector in a consistent alignment relative to the magnetic field?

Experiments were carried out to investigate the earlier prediction that prolonged exposure to long-wavelength (>500 nm) light would eliminate homing orientation by male Eastern red-spotted newts Notophthalmus viridescens. As in previous experiments, controls held in outdoor tanks under natural lighting conditions and tested in a visually uniform indoor arena under full-spectrum light were homeward oriented. As predicted, however, newts held under long-wavelength light and tested under either full-spectrum or long-wavelength light (>500 nm) failed to show consistent homeward orientation. The newts also did not orient with respect to the shore directions in the outdoor tanks in which they were held prior to testing. Unexpectedly, however, the newts exhibited bimodal orientation along a more-or-less `fixed' north-northeast—south-southwest magnetic axis. The orientation exhibited by newts tested under full-spectrum light was indistinguishable from that of newts tested under long-wavelength light, although these two wavelength conditions have previously been shown to differentially affect both shoreward compass orientation and homing orientation. To investigate the possibility that the `fixed-axis' response of the newts was mediated by a magnetoreception mechanism involving single-domain particles of magnetite, natural remanent magnetism (NRM) was measured from a subset of the newts. The distribution of NRM alignments with respect to the head—body axis of the newts was indistinguishable from random. Furthermore, there was no consistent relationship between the NRM of individual newts and their directional response in the overall sample. However, under full-spectrum, but not long-wavelength, light, the alignment of the NRM when the newts reached the 20 cm radius criterion circle in the indoor testing arena (estimated by adding the NRM alignment measured from each newt to its magnetic bearing) was non-randomly distributed. These findings are consistent with the earlier suggestion that homing newts use the light-dependent magnetic compass to align a magnetite-based `map detector' when obtaining the precise measurements necessary to derive map information from the magnetic field. However, aligning the putative map detector does not explain the fixed-axis response of newts tested under long-wavelength light. Preliminary evidence suggests that, in the absence of reliable directional information from the magnetic compass (caused by the 90° rotation of the response of the magnetic compass under long-wavelength light), newts may resort to a systematic sampling strategy to identify alignment(s) of the map detector that yields reliable magnetic field measurements

Stroke and TIA survivors’ cognitive beliefs and affective responses regarding treatment and future stroke risk differentially predict medication adherence and categorised stroke risk

Cognitive beliefs and affective responses to illness and treatment are known to independently predict health behaviours. The purpose of the current study is to assess the relative importance of four psychological domains – specifically, affective illness, cognitive illness, affective treatment and cognitive treatment – for predicting stroke and transient ischemic attack (TIA) survivors’ adherence to stroke prevention medications as well as their objective, categorised stroke risk. We assessed these domains among stroke/TIA survivors (n = 600), and conducted correlation and regression analyses with concurrent and prospective outcomes to determine the relative importance of each cognitive and affective domain for adherence and stroke risk. As hypothesised, patients’ affective treatment responses explained the greatest unique variance in baseline and six-month adherence reports (8 and 5%, respectively, of the variance in adherence, compared to 1–3% explained by other domains). Counter to hypotheses, patients’ cognitive illness beliefs explained the greatest unique variance in baseline and six-month objective categorised stroke risk (3 and 2%, respectively, compared to 0–1% explained by other domains). Results indicate that domain type (i.e. cognitive and affective) and domain referent (illness and treatment) may be differentially important for providers to assess when treating patients for stroke/TIA. More research is required to further distinguish between these domains and their relative importance for stroke prevention

The Changing Practice of Bankruptcy Law: An Analysis of How Bankruptcy Practice Has Changed in the Last Decade

The practice of bankruptcy law has changed drastically over the last decade. An attorney starting out in the field in 2009 faces different issue than one who began in 1999. However, it’s not just the issues that come up with clients that make the practice so different, but the law of bankruptcy itself has changed. The economic downturn of the last eighteen months has changed the way the public views bankruptcy. The Bankruptcy Reform Act of 2005 and In re Bateman, a case decided in 2008, altered the landscape of bankruptcy practice forever. This article will walk through a decade of bankruptcy reform, from the points of view of an attorney practicing in 1999 and one practicing in 2009. The purpose of this article is to provide a practical review of the new bankruptcy laws and their impact on how attorneys should practice in today’s bankruptcy world. Through a discussion of the economic climate, legal reform, and the social reform surrounding bankruptcy, we hope to educate today’s attorneys not only of the present state of the law, but the future of bankruptcy practice as well

Improved Constraints on Isotropic Shift and Anisotropies of the Speed of Light using Rotating Cryogenic Sapphire Oscillators

We demonstrate that Michelson-Morley tests, which detect direction-dependent anisotropies in the speed of light, can also be used to place limits upon isotropic deviations of the vacuum speed of light from $c$, as described by the photon sector Standard Model Extension (SME) parameter $\tilde{\kappa}_{tr}$. A shift in the speed of light that is isotropic in one inertial frame implies anisotropic shifts in others. Using observer Lorentz covariance, we derive the time-dependent variations in the relative resonance frequencies of a pair of electromagnetic resonators that would be generated by such a shift in the rest frame of the Sun. A new analysis of a recent experimental test of relativity using this result constrains $\tilde{\kappa}_{tr}$ with a precision of $7.4\times10^{-9}$. This represents the first constraint on $\tilde{\kappa}_{tr}$ by a Michelson-Morley experiment and the first analysis of a single experiment to simultaneously set limits on all nine non-birefringent terms in the photon sector of the SME

An Exploratory Study of Suboxone (Buprenorphine/ Naloxone) Film Splitting: Cutting Methods, Content Uniformity, and Stability

Suboxone films are U.S. Food and Drug Administration approved to treat opioid dependence. While the package insert states that films should not be cut, physicians often prescribe film fractions for treatment and tapering. There is no data to support this practice, and this study was initiated to evaluate cutting methods, content uniformity, and stability of split films. Suboxone 8-mg buprenorphine/2-mg naloxone films were split using four methods: 1) ruler/razor cut, 2) scissor cut, 3) fold/rip, and 4) fold/scissor cut. United States Pharmacopeia Chapter \u3c905\u3e was used to evaluate the weight variation and content uniformity of split films. The stability of split films stored in polybags was evaluated over 7 days. A stability-indicating high-performance liquid chromatography method was used for content uniformity and stability evaluation. The weight variation results were acceptable for the half films from all four cutting methods, but this was not true for the quarter films. The method of ruler/razor cut was determined most favorable and used for the content uniformity test. Based on the high-performance liquid chromatography results, the half films from the ruler/razor cut method met the passing criteria of United States Pharmacopeia Chapter \u3c905\u3e with acceptance values of 9.8 to 10.4 for buprenorphine and 8.4 to 11.5 for naloxone (≤15 is considered passing). The stability results indicated that both actives retained \u3e97.7% of initial strength. Four cutting methods were found to be acceptable for splitting Suboxone films into half but not quarter fractions. The half films from the ruler/razor cut method also passed United States Pharmacopeia Chapter \u3c905\u3e content uniformity test. Both actives remained stable for 7 days when the half films were stored in polybags at room temperature

Transforming NICU Care to Provide Comprehensive Family Support

AbstractParents whose babies are admitted to neonatal intensive care units (NICUs) are subject to a variety of stresses that increase their risk to suffer from postpartum depression, posttraumatic stress disorder, and anxiety disorders. Parental distress can adversely impact parent-infant bonding, which in turn can lead to worse outcomes for the child. An interdisciplinary workgroup convened by the National Perinatal Association (NPA) has published recommendations for psychosocial support of NICU parents. This article presents nurses, who are key players in quality improvement (QI) initiatives in their hospitals, with a QI approach to transforming care in NICUs to provide comprehensive family support using the NPA recommendations as a road map

Interfacial interaction of water and silicate minerals

This program has relation to the categories of the nature of water and water cycle. In particular, the study deals with the basic interactions of water with the surface of minerals. The program has direct application to the areas groundwater and water in soils in the category of water cycle. We have investigated the problems of bonding and migration of water molecules upon sheet structures of mineral samples. The minerals used in the program have surface structures that are similar and in some cases identical to those found in clays. This study focused upon the molecular level rather than on the bulk property level of capillarity and hydraulics. The interlayer binding energy of selenite was measured. The measurements were made within a test chamber in which the temperature and atmospheric pressure of the sample could be controlled and monitored. There was found to be no significant difference in the IBE of selenite in air (716 ergs/cm^2) and in vacuum (704 ergs/cm^2). There was found to be a slight difference in the low temperature IBE (768 ergs/cm^2) and the high temperature IBE (720 ergs/cm^2); however, this was attributable to effects of cold on the mechanism. It was concluded that the IBE io independent of atmospheric and temperature effects within the range of temperature and pressure studied.Project # A-057-MO Agreement # 14-31-0001-382