Interactions between the structural domains of the RNA replication proteins of plant-infecting RNA viruses

Brome mosaic virus (BMV), a positive-strand RNA virus, encodes two replication proteins: the 2a protein, which contains polymerase-like sequences, and the 1a protein, with N-terminal putative capping and C-terminal helicase-like sequences. These two proteins are part of a multisubunit complex which is necessary for viral RNA replication. We have previously shown that the yeast two-hybrid assay consistently duplicated results obtained from in vivo RNA replication assays and biochemical assays of protein-protein interaction, thus permitting the identification of additional interacting domains. We now map an interaction found to take place between two 1a proteins. Using previously characterized 1a mutants, a perfect correlation was found between the in vivo phenotypes of these mutants and their abilities to interact with wild-type 1a (wt1a) and each other. Western blot analysis revealed that the stabilities of many of the noninteracting mutant proteins were similar to that of wt1a. Deletion analysis of 1a revealed that the N-terminal 515 residues of the 1a protein are required and sufficient for 1a-1a interaction. This intermolecular interaction between the putative capping domain and itself was detected in another tripartite RNA virus, cucumber mosaic virus (CMV), suggesting that the 1a-1a interaction is a feature necessary for the replication of tripartite RNA viruses. The boundaries for various activities are placed in the context of the predicted secondary structures of several 1a-like proteins of members of the alphavirus-like superfamily. Additionally, we found a novel interaction between the putative capping and helicase-like portions of the BMV and CMV 1a proteins. Our cumulative data suggest a working model for the assembly of the BMV RNA replicase.     

Concept-Based Method for Extracting Valid Subsets from an EXPRESS Schema

An EXPRESS schema is a data schema defined in EXPRESS, an international standard language for defining product data schemas. This technical paper proposes and formally defines a set of conditions for generating a minimum valid subset of an EXPRESS schema corresponding to a concept, where a concept is a general idea and a subset is a partial model of a data schema. We introduce a notion of “minimal set” to define the relationships between a subset and other subsets, and also between a subset and concepts. A minimal set is the smallest complete subset of a schema that corresponds to a concept. Using IFC, an international standard data model for the architecture, engineering, and construction industry, the proposed conditions have been implemented in a software application developed for extracting subsets from the IFC schema matching the concepts. A number of examples are demonstrated.     

Effects of TCDD on Ah receptor, ARNT, EGF, and TGF-alpha expression in embryonic mouse urinary tract

Lipoprotein(a) [Lp(a)] biogenesis was examined in primary cultures of hepatocytes isolated from mice transgenic for both human apolipoprotein(a) [apo(a)] and human apoB. Steady-state and pulse-chase labeling experiments demonstrated that newly synthesized human apo(a) had a prolonged residence time (approximately 60 min) in the endoplasmic reticulum (ER) before maturation and secretion. Apo(a) was inefficiently secreted by the hepatocytes and a large portion of the protein was retained and degraded intracellularly. Apo(a) exhibited a prolonged and complex folding pathway in the ER, which included incorporation of apo(a) into high molecular weight, disulfide-linked aggregates. These folding characteristics could account for long ER residence time and inefficient secretion of apo(a). Mature apo(a) bound via its kringle domains to the hepatocyte cell surface before appearing in the culture medium. Apo(a) could be released from the cell surface by apoB-containing lipoproteins. These studies are consistent with a model in which the efficiency of post-translational processing of apo(a) strongly influences human plasma Lp(a) levels, and suggest that cell surface assembly may be one pathway of human Lp(a) production in vivo. Transgenic mouse hepatocytes thus provide a valuable model system with which to study factors regulating human Lp(a) biogenesis.     

Environment-dependent tight-binding potential model

The yellow-pigmented bacterium isolated from a ditch was a gram negative rod with a G+C content of 63 mol%, and was classified in the genus Sphingomonas. Electron microscopy revealed that the bacterial cell surface was covered with many large plaits. When grown in a medium containing a polysaccharide as an essential nutrient, a pit of 0.02-0.1 micrometers in diameter was formed on the cell surface, and a thin section showed the rearrangement of the plaits and the presence of a region where the cell membrane sinks into the cytosol. The dependence of the pit formation on the presence of macromolecule may predict the existence of a direct uptake mechanism for macromolecules through a mouth-like pit, possibly in endocytosis fashion. The confirmation of the pit structure is the first such finding in the history of microbiology and may provide a new insight into the cell morphology and biochemistry of macromolecule transport in microbial cell system.     

Mathematical modeling of tornadoes and squall storms

Recent advances in modeling of tornadoes and twisters consist of significant achievements in mathematical calculation of occurrence and evolution of a violent F5-class tornado on the Fujita scale, and four-dimensional mathematical modeling of a tornado with the fourth coordinate time multiplied by its characteristic velocity. Such a tornado can arise in a thunderstorm supercell filled with turbulent whirlwinds. A theory of the squall storms is proposed. The squall storm is modeled by running perturbation of the temperature inversion on the lower boundary of cloudiness. This perturbation is induced by the action of strong, hurricane winds in the upper and middle troposphere, and looks like a running solitary wave (soliton); which is developed also in a field of pressure and velocity of a wind. If a soliton of a squall storm gets into the thunderstorm supercell then this soliton is captured by supercell. It leads to additional pressure fall of air inside a storm supercell and stimulate amplification of wind velocity here. As a result, a cyclostrophic balance inside a storm supercell generates a tornado. Comparison of the radial distribution of wind velocity inside a tornado calculated by using the new formulas and equations with radar observations of the wind velocity inside Texas Tornado Dummit in 1995 and inside the 3 May 1999 Oklahoma City Tornado shows good correspondence.     

Dynamic interaction between a coherent precipitate and an edge dislocation

Dynamic interaction between a coherent precipitate and an edge dislocation is analyzed by means of a discrete atom method, which is based on classical statistical mechanics and linear elasticity. Precipitates having a dilatational misfit strain and elastic constants different from those of the matrix phase are treated in anisotropic elastic systems under a plane strain condition. A coherent interface transforms into a semicoherent one by nucleating dipolar dislocations at a stress concentration in a coherent precipitate. One of the dipolar dislocations glides along the precipitate-matrix interface to become a misfit dislocation, and the other slips into the matrix phase to become a lattice dislocation. In accordance with continuum elasticity, a coherent particle with a positive misfit strain migrates to the tension side of an edge dislocation, whereas a particle with a negative misfit diffuses to the region of compression. Morphological change is, however, caused by the dislocation as the particle tries to capitalize on the dislocation stress field, and the particle shape depends on its stiffness and elastic anisotropy. Under an applied shear, a hard coherent particle with a positive misfit strain is sheared along the shear direction, but a soft particle responds in the opposite direction. Elastic interaction between a coherent particle and an edge dislocation can be so strong that the particle-dislocation complex remains coupled even at a high shear strain applied to the system. Some composite applied stresses can cause an edge dislocation to split into two partials. One of the partials, a glissile component, is found to engage actively in the morphological evolution of a particle during a diffusional relaxation. This article is based on a presentation made in the symposium “Kinetically Determined Particle Shapes and the Dynamics of Solid:Solid Interfaces,” presented at the October 1996 Fall meeting of TMS/ASM in Cincinnati, Ohio, under the auspices of the ASM Phase Transformations Committee.     

Amyloid beta-peptide spin trapping. II: Evidence for decomposition of the PBN spin adduct

We investigated the effect of lipoprotein(a) (Lp(a)) on proliferation of human arterial smooth muscle cells (SMCs) and its mechanisms of action. Low density lipoprotein (LDL), Lp(a) and apolipoprotein(a) (apo(a)) significantly stimulated the proliferation of SMCs. Lp(a) and apo(a) reduced the amount of active transforming growth factor-beta (TGF-beta) with the mink lung epithelial cell bioassay, however LDL had no effect. Lp(a), but not apo(a), significantly stimulated the proliferation of SMCs even in the presence of a neutralizing antibody for TGF-beta. Our results suggest that Lp(a) stimulates the proliferation of SMCs via apo(a)-induced inhibition of TGF-beta activation and stimulation of SMCs by the LDL-particle of Lp(a).     

Apolipoprotein(a) binds via its C-terminal domain to the protein core of the proteoglycan decorin. Implications for the retention of lipoprotein(a) in atherosclerotic lesions

Although it is known that lipoprotein(a) (Lp(a)) binds to proteoglycans, the mechanism for this binding has not been fully elucidated. In order to shed light on this subject, we examined the interactions of decorin, a proteoglycan with a well defined protein core and a single glycosaminoglycan (GAG) chain, with Lp(a) and derivatives, namely Lp(a) deprived of apo(a), or Lp(a-), free apo(a), and the two main proteolytic fragments, F1 and F2. By circular dichroism criteria, the decorin preparations used had the same secondary structure as that previously reported for native decorin. Authentic low density lipoprotein from the same human donor was used as a control. In a solid phase system, Lp(a-)and low density lipoprotein bound to decorin in a comparable manner. This binding required Ca2+/Mg2+ ions, was lysine-mediated, and was markedly decreased in the presence of GAG-depleted decorin, suggesting the ionic nature of the interaction likely involving apoB100 and the GAG component of decorin. Free apo(a) also bound to decorin; however, the binding was neither cation-dependent nor lysine-mediated, unaffected by sialic acid depletion of apo(a), and markedly decreased when either reduced and alkylated apo(a) or reduced and alkylated decorin was used in the assay. Of note, the binding of apo(a) was unaffected when it was incubated with a spectrally native decorin that had been renatured from either 4 M guanidine hydrochloride by extensive dialysis or cooled from 65 to 25 degrees C. On the other hand, the binding significantly increased when decorin was depleted of GAGs, which by themselves had no affinity for apo(a). The binding of apo(a) to the decorin protein core was also elicited by the C-terminal domain of apo(a), and it was favored by high NaCl concentrations, 1 to 2 M. No binding was exhibited by the N-terminal domain accounting for the lack of effect of apo(a) size polymorphism on the binding. In the case of whole Lp(a), the binding to immobilized decorin was mostly GAG-dependent and ionic in nature. A minor contribution by apo(a) was detected when GAG-depleted decorin was used in the assay. Our results indicate that the binding of Lp(a) to decorin involves interactions both electrostatic (apoB100-GAG) and hydrophobic (apo(a)-decorin protein core), and that the binding of apo(a) requires decorin protein core to be in its native state.     

Transition to Self-Similarity of Diffusion of Tracer in Turbulent Patch

A mixing of a passive tracer inside a turbulent patch generated by a localized short-time perturbation is studied numerically and analytically. Two kinds of an initial distribution of a tracer are considered: two-layer and continuous with constant gradient. For the turbulent patch shaped as a layer, it is shown that, regardless of details of initial distributions of a turbulent energy and dissipation, a tracer concentration evolves to self-similar regimes as time elapses. Analytical self-similar solutions to turbulent diffusion equations are found for three symmetric shapes of a turbulent patch: layer, cylinder, and sphere. Distributions of the concentration inside a patch are found to be substantially nonuniform, with a typical ratio of a concentration gradient in the middle of a patch to its initial value of about 0.5.     

The arterial blood supply of the pancreas: a review. IV. The anterior inferior and posterior pancreaticoduodenal aa., and minor sources of blood supply for the head of the pancreas. An anatomical review and radiologic study

The present article is the fourth part of a comprehensive review of the arterial blood supply of the pancreas and completes the study of the arterial vascularization of the pancreatic head dealing with the anterior inferior and posterior inferior pancreaticoduodenal aa. and with some minor sources of blood supply not involving the classical system of the pancreaticoduodenal arches. The aim of this review is to summarise the anatomical studies, starting from Haller's reports, and to supply, as far as possible with original material, angiographic evidence for the classic anatomical concepts. For this purpose, 1015 selective angiographs (celiac trunk and its branches, superior mesenteric a.) were taken from the angiographic archives of the Institutes of Radiology of Siena, Rome (Catholic University), and Perugia. These demonstrated the anterior inferior pancreaticoduodenal a., present in most instances, as arising from the inferior pancreaticoduodenal a., from a common trunk with the posterior inferior pancreaticoduodenal a. and the 1st jejunal a., from the 1st jejunal a. or from the superior mesenteric a.; on the other hand, the posterior inferior pancreaticoduodenal a. was more variable, originating from the inferior pancreaticoduodenal a., from a common trunk with the anterior inferior pancreaticoduodenal a. and the 1st jejunal a., from the superior mesenteric a., from the dorsal pancreatic a., or from a right accessory hepatic a. coming from the superior mesenteric a. In addition, minor branches to the head of the pancreas arose from the gastroduodenal a., the dorsal pancreatic a., the common hepatic a. and the inferior right phrenic a. Other origins of the inferior pancreaticoduodenal aa. previously reported, but not angiographically detectable with certainty, as well as further minor sources of blood supply to the head of the pancreas, have been listed. The differing opinions regarding the incidence of the various ways the inferior pancreaticoduodenal aa. arise are discussed and an attempt is made to explain the variability of the vascular anatomy of the pancreatic head on embryologic grounds.     

Structural basis for the pathophysiology of lipoprotein(a) in the athero-thrombotic process

Lipoprotein Lp(a) is a major and independent genetic risk factor for atherosclerosis and cardiovascular disease. The essential difference between Lp(a) and low density lipoproteins (LDL) is apolipoprotein apo(a), a glycoprotein structurally similar to plasminogen, the precursor of plasmin, the fibrinolytic enzyme. This structural homology endows Lp(a) with the capacity to bind to fibrin and to membrane proteins of endothelial cells and monocytes, and thereby to inhibit plasminogen binding and plasmin generation. The inhibition of plasmin generation and the accumulation of Lp(a) on the surface of fibrin and cell membranes favor fibrin and cholesterol deposition at sites of vascular injury. Moreover, insufficient activation of TGF-beta due to low plasmin activity may result in migration and proliferation of smooth muscle cells into the vascular intima. These mechanisms may constitute the basis of the athero-thrombogenic mode of action of Lp(a). It is currently accepted that this effect of Lp(a) is linked to its concentration in plasma. An inverse relationship between Lp(a) concentration and apo(a) isoform size, which is under genetic control, has been documented. Recently, it has been shown that inhibition of plasminogen binding to fibrin by apo(a) is also inversely associated with isoform size. Specific point mutations may also affect the lysine-binding function of apo(a). These results support the existence of functional heterogeneity in apolipoprotein(a) isoforms and suggest that the predictive value of Lp(a) as a risk factor for vascular occlusive disease would depend on the relative concentration of the isoform with the highest affinity for fibrin.     

Oxidation of apolipoprotein(a) inhibits kringle-associated lysine binding: the loss of intrinsic protein fluorescence suggests a role for tryptophan residues in the lysine binding site

Lipoprotein(a) [Lp(a)] is a low-density lipoprotein complex consisting of apolipoprotein(a) [apo(a)] disulfide-linked to apolipoprotein B-100. Lp(a) has been implicated in atherogenesis and thrombosis through the lysine binding site (LBS) affinity of its kringle domains. We have examined the oxidative effect of 2,2'-azobis-(amidinopropane) HCl (AAPH), a mild hydrophilic free radical initiator, upon the ability of Lp(a) and recombinant apo(a), r-apo(a), to bind through their LBS domains. AAPH treatment caused a time-dependent decrease in the number of functional Lp(a) or r-apo(a) molecules capable of binding to fibrin or lysine-Sepharose and in the intrinsic protein fluorescence of both Lp(a) and r-apo(a). The presence of a lysine analogue during the reaction prevented the loss of lysine binding and provided a partial protection from the loss of tryptophan fluorescence. The partial protection of fluorescence by lysine analogues was observed in other kringle-containing proteins, but not in proteins lacking kringles. No significant aggregation, fragmentation, or change in conformation of Lp(a) or r-apo(a) was observed as assessed by native or SDS-PAGE, light scattering, retention of antigenicity, and protein fluorescence emission spectra. Our results suggest that AAPH destroys amino acids in the kringles of apo(a) that are essential for lysine binding, including one or more tryptophan residues. The present study, therefore, raises the possibility that the biological roles of Lp(a) may be mediated by its state of oxidation, especially in light of our previous study showing that the reductive properties of sulfhydryl-containing compounds increase the LBS affinity of Lp(a) for fibrin.     

The well of loneliness, or the gospel according to Radclyffe Hall

Radclyffe Hall's 1928 novel, The Well of Loneliness, is repeatedly described as a "bible" of lesbian literature. The novel itself repeatedly alludes to biblical stories, especially the story of Christ. Yet there has been little sustained analysis of the biblical language of the novel. Most feminist and lesbian critics have dismissed the biblical allusions and language as unfortunate and politically regressive; religious critics have ignored the novel. This essay reexamines the biblical nature of the novel, especially its portrayal of the lesbian Stephen Gordon as a Christ figure. The study further claims a creative and interventionary power in Hall's use of biblical narratives and tropes, a power traceable in public reception to the novel and in courtroom reactions to the use of spiritual language in a text about lesbianism. By writing the life of a lesbian as a kind of gospel of inversion, Hall turns a language of condemnation into a language of validation, making her use of biblical language a kind of Foucauldian "reverse discourse." The novel's power lies in its portrayal of a lesbian messiah, and in its joining of sexological and religious discourses.     

Complex injuries of the dorsum of the hand. Therapeutic reflections apropos of 2 cases

We report 2 similar cases of severe injury of the dorsum of the hand, both treated in emergency. FIRST CASE: A 37-year-old right handed male truck driver was admitted with a complex severe injury of the dorsum of his right hand following a traffic accident. He had a combined large defect involving skin, tendons and bone. A complex reconstruction was performed using a massive iliac crest allograft, a tendon graft and a free serratus anterior flap in a one stage procedure, eight hours after the injury. A skin graft was done later. Two years later the functional and esthetic results are good. SECOND CASE: A 37-year-old woman was admitted with a severe injury of the dorsum of her right hand following a traffic accident. She had a large combined defect involving skin, tendon and bone. A complex reconstruction was carried out using a large iliac crest autograft, a multiple tendon graft and a free latissimus dorsi flap, in one stage 6 hours after the trauma. Eight months later the functional result is partial but useful for the daily activities of the patient.     

Decubitus prevention in theory and practice--a study structure with the VIPS model

In 1990, the Bureau of Dental Health Services of the New York City Department of Health launched a major initiative to modernize a network of school-based dental clinics located throughout the city. Since 1913, the bureau has provided dental care to public school children; however, the clinics were not properly maintained or upgraded, and were in a state of disrepair and obsolescence. Anticipating that the survival of the program was in question, the school program was converted to a fleet of state-of-the-art portable dental clinics permitting targeting of underserved, high-risk poor and immigrant populations. Demographics had changed dramatically over the years; the program could now situate services where they were needed most, and provide a broader array of care where access was a problem. This paper presents a six-year analysis of the program and builds a strong case to show that a portable delivery system can equal or in many ways surpass the effectiveness and capabilities of a fixed-state approach.     

Probability of Project Completion Using Stochastic Project Scheduling Simulation

This paper introduces a software, Stochastic Project Scheduling Simulation (SPSS), developed to measure the probability to complete a project in a certain time specified by the user. To deliver a project by a completion date committed to in a contract, a number of activities need to be carried out. The time that an entire project takes to complete and the activities that determine total project duration are always questionable because of the randomness and stochastic nature of the activities’ durations. Predicting a project completion probability is valuable, particularly at the time of bidding. The SPSS finds the longest path in a network and runs the network a number of times specified by the user and calculates the stochastic probability to complete the project in the specified time. The SPSS can be used by a contractor: (1) to predict the probability to deliver the project in a given time frame and (2) to assess its capabilities to meet the contractual requirement before bidding. The SPSS can also be used by a construction owner to quantify and analyze the risks involved in the schedule. The benefits of the tool to researchers are: (1) to solve program evaluation and review technique problems; (2) to complement Monte Carlo simulation by applying the concept of project network modeling and scheduling with probabilistic and stochastic activities via a web based Java Simulation which is operateable over the Internet, and (3) to open a way to compare a project network having different distribution functions.     

Nitric oxide synthase inhibition in a spontaneously hypertensive rat model of diabetic nephropathy

Squamous odontogenic tumour is a benign odontogenic tumour composed of a well-differentiated squamous epithelium immersed in a fibrous connective tissue stroma. It is a rare tumour and a recent literature review yielded only 36 cases. Two cases of squamous odontogenic tumour are presented, 1 located in the maxilla and the other in the mandible: 1 of these cases showed a periodontal involvement. The radiographic picture was fairly characteristic in 1 case, with a radiolucent lesion between the roots of the second mandibular premolar and the first molar, while, in the other case, it was possible to observe the presence of a lesion located at the apex of a molar. The tumours were enucleated, and no recurrences were observed after 5 years.