The low molecular weight (LMW) isoforms of cyclin E provide a novel mechanism of cell cycle deregulation

Cyclin E, in complex with cyclin dependent kinase 2 (CDK2), is a positive regulator of G1 to S phase progression of the cell cycle. Deregulation of G1/S phase transition occurs in the majority of tumors. Cyclin E is overexpressed and post-translationally generates low molecular weight (LMW) isoforms in breast cancer, but not normal cells. Such alteration of cyclin E is linked to poor prognosis. Therefore, we hypothesized that the LMW isoforms of cyclin E provide a novel mechanism of cell cycle de-regulation in cancer cells. Insect cell expression system was used to explore the biochemical properties of the cyclin E isoforms. Non-tumorigenic (76NE6) and tumorigenic (T47D) mammary epithelial cells transfected with the cyclin E isoforms and breast tumor tissue endogenously expressing the LMW isoforms were used to study the biologic consequences of the LMW isoforms of cyclin E. All model systems studied show that the LMW forms (compared to full-length cyclin E) have increased kinase activity when partnered with CDK2. Increases in the percentage of cells in S phase and colony formation were also observed after overexpression of LMW compared to full-length cyclin E. The LMW isoforms of cyclin E utilize several mechanisms to attain their hyper-activity. They bind CDK2 more efficiently, and are resistant to inhibition by cyclin dependent kinase inhibitors (CKIs) as compared to full-length cyclin E. In addition, the LMW isoforms sequester the CKIs from full-length cyclin E abrogating the overall negative regulation of cyclin E. Despite their correlation with adverse biological consequences, the direct role of the LMW isoforms of cyclin E in mediating tumorigenesis remained unanswered. Subsequent to LMW cyclin E expression in 76NE6 cells, they lose their ability to enter quiescence and exhibit genomic instability, both characteristic of a tumor cell phenotype. Furthermore, injection of 76NE6 cells overexpressing each of the cyclin E isoforms into the mammary fat pad of nude mice revealed that the LMW isoforms of cyclin E yield tumors, whereas the full-length cyclin E does not. In conclusion, the LMW isoforms of cyclin E utilize several mechanisms to acquire a hyperactive phenotype that results in deregulation of the cell cycle and initiates the tumorigenic process in otherwise non-transformed mammary epithelial cells. ^

The Comal County Needs Assessment Youth Survey

A crucial link in preserving and protecting the future of our communities resides in maintaining the health and well being of our youth. While every member of the community owns an opinion regarding where to best utilize monies for prevention and intervention, the data to support such opinion is often scarce. In an effort to generate data-driven indices for community planning and action, the United Way of Comal County, Texas partnered with the University Of Texas - Houston Health Science Center, School Of Public Health to accomplish a county-specific needs assessment. A community-based participatory research emphasis utilizing the Mobilization for Action through Planning and Partnership (MAPP) format developed by the National Association of City and County Health Officials (NACCHO) was implemented to engage community members in identifying and addressing community priorities. The single greatest area of consensus and concern identified by community members was the health and well being of the youth population. Thus, a youth survey, targeting these specific areas of community concern, was designed, coordinated and administered to all 9-11th grade students in the county. 20% of the 3,698 completed surveys (72% response rate) were randomly selected for analysis. These 740 surveys were coded and scanned into an electronic survey database. Statistical analysis provided youth-reported data on the status of the multiple issues affecting the health and well being of the community's youth. These data will be reported back to the community stakeholders, as part of the larger Comal County Needs Assessment, for the purposes of community planning and action. Survey data will provide community planners with an awareness of the high risk behaviors and habit patterns amongst their youth. This knowledge will permit more effective targeting of the means for encouraging healthy behaviors and preventing the spread of disease. Further, the community-oriented, population-based nature of this effort will provide answers to questions raised by the community and will provide an effective launching pad for the development and implementation of targeted, preventive health strategies. ^

2012-2014 sedimentation data for the Congaree River floodplain, SC, USA

This dataset contains sedimentation data that was collected for the Congaree River floodplain, SC, USA. The study sites were located at floodplain positions with varying flood frequencies and geomorphic positions. Flood frequency was determined using hydrological statistics based on data acquired from USGS gage #02169500 for the Congaree River; sites are classified as having high, intermediate, or low flood frequency. The geomorphic position classification was determined using field observations; sites are classified as abandoned channel, ridge, river margin, and swale positions. Sediment deposition was measured in centimeters and reflects the amount of sediment that was deposited on a feldspar clay pad that was installed at each of the sampling sites. The feldspar pads were installed in the summer of 2012 and cored and measured for deposition amounts in the summer of 2014. In the summer of 2012, soil samples were collected at each of the sampling sites. Each soil sample was analyzed for soil texture (percent sand, silt, and clay), major and micro-nutrients (parts per million), percent organic matter, and pH.

Order 10 4 speedup in global linear instability analysis using matrix formation

A unified solution framework is presented for one-, two- or three-dimensional complex non-symmetric eigenvalue problems, respectively governing linear modal instability of incompressible fluid flows in rectangular domains having two, one or no homogeneous spatial directions. The solution algorithm is based on subspace iteration in which the spatial discretization matrix is formed, stored and inverted serially. Results delivered by spectral collocation based on the Chebyshev-Gauss-Lobatto (CGL) points and a suite of high-order finite-difference methods comprising the previously employed for this type of work Dispersion-Relation-Preserving (DRP) and Padé finite-difference schemes, as well as the Summationby- parts (SBP) and the new high-order finite-difference scheme of order q (FD-q) have been compared from the point of view of accuracy and efficiency in standard validation cases of temporal local and BiGlobal linear instability. The FD-q method has been found to significantly outperform all other finite difference schemes in solving classic linear local, BiGlobal, and TriGlobal eigenvalue problems, as regards both memory and CPU time requirements. Results shown in the present study disprove the paradigm that spectral methods are superior to finite difference methods in terms of computational cost, at equal accuracy, FD-q spatial discretization delivering a speedup of ð (10 4). Consequently, accurate solutions of the three-dimensional (TriGlobal) eigenvalue problems may be solved on typical desktop computers with modest computational effort.

AVALIAÇÃO IN VITRO DA RESISTÊNCIA DA COLAGEM DE BRAQUETES LINGUAIS COM DIFERENTES ESPESSURAS DE RESINA

Este estudo avaliou a colagem de braquetes linguais nas faces linguais de pré-molares superiores com diferentes espessuras de PADs, confeccionados com a resina fotopolimerizável Transbond XT nas espessuras de 1,0 mm e 2,0 mm e comparando-os com a menor espessura possível. Foi utilizado o adesivo Sondhi Rapid Set para a colagem indireta. Avaliou-se a força de resistência sob cisalhamento cinco minutos após a colagem em uma máquina de ensaios mecânicos Kratos, com velocidade de cruzeta de 1,0 mm/min. A força média da resistência da colagem sob cisalhamento para o Grupo I foi de 9,69 MPa (DP 4,02 MPa), para o Grupo II foi de 6,15 MPa (DP 2,69 MPa) e para o Grupo III foi de 5,73 MPa (DP 1,62 MPa). O Grupo I, com menor espessura do PAD, apresentou força de resistência da colagem sob cisalhamento significativamente maior do que os Grupos II e III (PADs com 1,0 e 2,0 mm respectivamente). Estes por sua vez, não apresentaram diferença estatisticamente significante ao nível de p<0,05. O índice 1 de Adesivo Remanescente predominou nos Grupos I e II, caracterizando um maior número de fraturas do tipo adesiva. No Grupo 3 predominou o Índice 2, com fraturas do tipo coesiva.(AU)

AVALIAÇÃO IN VITRO DA RESISTÊNCIA DA COLAGEM DE BRAQUETES LINGUAIS COM DIFERENTES ESPESSURAS DE RESINA

Este estudo avaliou a colagem de braquetes linguais nas faces linguais de pré-molares superiores com diferentes espessuras de PADs, confeccionados com a resina fotopolimerizável Transbond XT nas espessuras de 1,0 mm e 2,0 mm e comparando-os com a menor espessura possível. Foi utilizado o adesivo Sondhi Rapid Set para a colagem indireta. Avaliou-se a força de resistência sob cisalhamento cinco minutos após a colagem em uma máquina de ensaios mecânicos Kratos, com velocidade de cruzeta de 1,0 mm/min. A força média da resistência da colagem sob cisalhamento para o Grupo I foi de 9,69 MPa (DP 4,02 MPa), para o Grupo II foi de 6,15 MPa (DP 2,69 MPa) e para o Grupo III foi de 5,73 MPa (DP 1,62 MPa). O Grupo I, com menor espessura do PAD, apresentou força de resistência da colagem sob cisalhamento significativamente maior do que os Grupos II e III (PADs com 1,0 e 2,0 mm respectivamente). Estes por sua vez, não apresentaram diferença estatisticamente significante ao nível de p<0,05. O índice 1 de Adesivo Remanescente predominou nos Grupos I e II, caracterizando um maior número de fraturas do tipo adesiva. No Grupo 3 predominou o Índice 2, com fraturas do tipo coesiva.(AU)

Suppression of breast cancer growth and metastasis by a serpin myoepithelium-derived serine proteinase inhibitor expressed in the mammary myoepithelial cells

A serpin was identified in normal mammary gland by differential cDNA sequencing. In situ hybridization has detected this serpin exclusively in the myoepithelial cells on the normal and noninvasive mammary epithelial side of the basement membrane and thus was named myoepithelium-derived serine proteinase inhibitor (MEPI). No MEPI expression was detected in the malignant breast carcinomas. MEPI encodes a 405-aa precursor, including an 18-residue secretion signal with a calculated molecular mass of 46 kDa. The predicted sequence of the new protein shares 33% sequence identity and 58% sequence similarity to plasminogen activator inhibitor (PAI)-1 and PAI-2. To determine whether MEPI can modulate the in vivo growth and progression of human breast cancers, we transfected a full-length MEPI cDNA into human breast cancer cells and studied the orthotopic growth of MEPI-transfected vs. control clones in the mammary fat pad of athymic nude mice. Overexpression of MEPI inhibited the invasion of the cells in the in vitro invasion assay. When injected orthotopically into nude mice, the primary tumor volumes, axillary lymph node metastasis, and lung metastasis were significantly inhibited in MEPI-transfected clones as compared with controls. The expression of MEPI in myoepithelial cells may prevent breast cancer malignant progression leading to metastasis.

Solution of the pulse width modulation problem using orthogonal polynomials and Korteweg-de Vries equations

The mathematical underpinning of the pulse width modulation (PWM) technique lies in the attempt to represent “accurately” harmonic waveforms using only square forms of a fixed height. The accuracy can be measured using many norms, but the quality of the approximation of the analog signal (a harmonic form) by a digital one (simple pulses of a fixed high voltage level) requires the elimination of high order harmonics in the error term. The most important practical problem is in “accurate” reproduction of sine-wave using the same number of pulses as the number of high harmonics eliminated. We describe in this paper a complete solution of the PWM problem using Padé approximations, orthogonal polynomials, and solitons. The main result of the paper is the characterization of discrete pulses answering the general PWM problem in terms of the manifold of all rational solutions to Korteweg-de Vries equations.

Integrin activation controls metastasis in human breast cancer

Metastasis is the primary cause of death in human breast cancer. Metastasis to bone, lungs, liver, and brain involves dissemination of breast cancer cells via the bloodstream and requires adhesion within the vasculature. Blood cell adhesion within the vasculature depends on integrins, a family of transmembrane adhesion receptors, and is regulated by integrin activation. Here we show that integrin αvβ3 supports breast cancer cell attachment under blood flow conditions in an activation-dependent manner. Integrin αvβ3 was found in two distinct functional states in human breast cancer cells. The activated, but not the nonactivated, state supported tumor cell arrest during blood flow through interaction with platelets. Importantly, activated αvβ3 was expressed by freshly isolated metastatic human breast cancer cells and variants of the MDA-MB 435 human breast cancer cell line, derived from mammary fat pad tumors or distant metastases in severe combined immunodeficient mice. Expression of constitutively activated mutant αvβ3D723R, but not αvβ3WT, in MDA-MB 435 cells strongly promoted metastasis in the mouse model. Thus breast cancer cells can exhibit a platelet-interactive and metastatic phenotype that is controlled by the activation of integrin αvβ3. Consequently, alterations within tumors that lead to the aberrant control of integrin activation are expected to adversely affect the course of human breast cancer.

Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors

The development of drugs for the control of tumor angiogenesis requires a simple, accurate, and economical assay for tumor-induced vascularization. We have adapted the orthotopic implantation model to angiogenesis measurement by using human tumors labeled with Aequorea victoria green fluorescent protein for grafting into nude mice. The nonluminous induced capillaries are clearly visible against the very bright tumor fluorescence examined either intravitally or by whole-body luminance in real time. The orthotopic implantation model of human cancer has been well characterized, and fluorescence shadowing replaces the laborious histological techniques for determining blood vessel density. Intravital images of orthotopically implanted human pancreatic tumors clearly show angiogenic capillaries at both primary and metastatic sites. A quantitative time course of angiogenesis was determined for an orthotopically growing human prostate tumor periodically imaged intravitally in a single nude mouse over a 19-day period. Whole-body optical imaging of tumor angiogenesis was demonstrated by injecting fluorescent Lewis lung carcinoma cells into the s.c. site of the footpad of nude mice. The footpad is relatively transparent, with comparatively few resident blood vessels, allowing quantitative imaging of tumor angiogenesis in the intact animal. Capillary density increased linearly over a 10-day period as determined by whole-body imaging. Similarly, the green fluorescent protein-expressing human breast tumor MDA-MB-435 was orthotopically transplanted to the mouse fat pad, where whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period. These powerful and clinically relevant angiogenesis mouse models can be used for real-time in vivo evaluation of agents inhibiting or promoting tumor angiogenesis in physiological microenvironments.

Biomechanics of the movable pretarsal adhesive organ in ants and bees

Hymenoptera attach to smooth surfaces with a flexible pad, the arolium, between the claws. Here we investigate its movement in Asian weaver ants (Oecophylla smaragdina) and honeybees (Apis mellifera).  When ants run upside down on a smooth surface, the arolium is unfolded and folded back with each step. Its extension is strictly coupled with the retraction of the claws. Experimental pull on the claw-flexor tendon revealed that the claw-flexor muscle not only retracts the claws, but also moves the arolium. The elicited arolium movement comprises (i) about a 90° rotation (extension) mediated by the interaction of the two rigid pretarsal sclerites arcus and manubrium and (ii) a lateral expansion and increase in volume. In severed legs of O. smaragdina ants, an increase in hemolymph pressure of 15 kPa was sufficient to inflate the arolium to its full size. Apart from being actively extended, an arolium in contact also can unfold passively when the leg is subject to a pull toward the body.  We propose a combined mechanical–hydraulic model for arolium movement: (i) the arolium is engaged by the action of the unguitractor, which mechanically extends the arolium; (ii) compression of the arolium gland reservoir pumps liquid into the arolium; (iii) arolia partly in contact with the surface are unfolded passively when the legs are pulled toward the body; and (iv) the arolium deflates and moves back to its default position by elastic recoil of the cuticle.

Extracellular Matrix Assembly in Diatoms (Bacillariophyceae)1 : III. Organization of Fucoglucuronogalactans within the Adhesive Stalks of Achnanthes longipes

Achnanthes longipes is a marine, biofouling diatom that adheres to surfaces via adhesive polymers extruded during motility or organized into structures called stalks that contain three distinct regions: the pad, shaft, and collar. Four monoclonal antibodies (AL.C1–AL.C4) and antibodies from two uncloned hybridomas (AL.E1 and AL.E2) were raised against the extracellular adhesives of A. longipes. Antibodies were screened against a hot-water-insoluble/hot-bicarbonate-soluble-fraction. The hot-water-insoluble/hot-bicarbonate-soluble fraction was fractionated to yield polymers in three size ranges: F1, ≥ 20,000,000 Mr; F2, ≅100,000 Mr; and F3, <10,000 Mr relative to dextran standards. The ≅100,000-Mr fraction consisted of highly sulfated (approximately 11%) fucoglucuronogalactans (FGGs) and low-sulfate (approximately 2%) FGGs, whereas F1 was composed of O-linked FGG (F2)-polypeptide (F3) complexes. AL.C1, AL.C2, AL.C4, AL.E1, and AL.E2 recognized carbohydrate complementary regions on FGGs, with antigenicity dependent on fucosyl-containing side chains. AL.C3 was unique in that it had a lower affinity for FGGs and did not label any portion of the shaft. Enzyme-linked immunosorbent assay and immunocytochemistry indicated that low-sulfate FGGs are expelled from pores surrounding the raphe terminus, creating the cylindrical outer layers of the shaft, and that highly sulfated FGGs are extruded from the raphe, forming the central core. Antibody-labeling patterns and other evidence indicated that the shaft central-core region is related to material exuded from the raphe during cell motility.

Systemic immunization with papillomavirus L1 protein completely prevents the development of viral mucosal papillomas.

Infection of mucosal epithelium by papillomaviruses is responsible for the induction of genital and oral warts and plays a critical role in the development of human cervical and oropharyngeal cancer. We have employed a canine model to develop a systemic vaccine that completely protects against experimentally induced oral mucosal papillomas. The major capsid protein, L1, of canine oral papillomavirus (COPV) was expressed in Sf9 insect cells in native conformation. L1 protein, which self-assembled into virus-like particles, was purified on CsCl gradients and injected intradermally into the foot pad of beagles. Vaccinated animals developed circulating antibodies against COPV and became completely resistant to experimental challenge with COPV. Successful immunization was strictly dependent upon native L1 protein conformation and L1 type. Partial protection was achieved with as little as 0.125 ng of L1 protein, and adjuvants appeared useful for prolonging the host immune response. Serum immunoglobulins passively transferred from COPV L1-immunized beagles to naive beagles conferred protection from experimental infection with COPV. Our results indicate the feasibility of developing a human vaccine to prevent mucosal papillomas, which can progress to malignancy.