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At least 631 records · Page 35Linked to original sources

Automatic detection of the boundary of the calcaneus from ultrasound parametric images using an active contour model; clinical assessment.

This paper presents a computerized method for automated detection of the boundary of the os calcis on in vivo ultrasound parametric images, using an active dynamic contour model. The initial contour, defined without user interaction, is an iso-contour extracted from the textural feature space. The contour is deformed through the action of internal and external forces, until stability is reached. The external forces, which characterize image features, are a combination of gray-level information and second-order textural features arising from local cooccurrence matrices. The broadband ultrasound attenuation (BUA) value is then averaged within the contour obtained. The method was applied to 381 clinical images. The contour was correctly detected in the great majority of the cases. For the short-term reproducibility study, the mean coefficient of variation was equal to 1.81% for BUA values and 4.95% for areas in the detected region. Women with osteoporosis had a lower BUA than age-matched controls (p = 0.0005). In healthy women, the age-related decline was -0.45 dB/MHz/yr. In the group of healthy post-menopausal women, years since menopause, weight and age were significant predictors of BUA. These results are comparable to those obtained when averaging BUA values in a small region of interest.

Adult↗

A practical way to diagnosis personality disorder: a proposal.

General criteria for the diagnosis of personality disorder are provided based on rating a few items describing four core features of personality disorder: (a) low Self-directedness, (b) low Cooperativeness, (c) low Affective Stability, and (d) low Self-transcendence. These core features correspond closely with the basic concept of personality disorder in DSM-IV and are based on specific items easy for clinicians to rate in a short time. Criteria are also provided for rating severity of personality disorganization and for subtyping based on a profile of three additional dimensions corresponding to core features of DSM-IV clusters A, B, and C. This approach should facilitate efficient screening in clinical practice, encourage an understanding of the development of comorbidity as a self-organizing process, and provide a theory-driven basis for therapeutic planning with drugs and psychotherapy.

Comorbidity↗

The P.F.C. sigma RP-F total knee arthroplasty: designed for improved performance.

The new P.F.C. Sigma RP-F (rotating-platform high-flexion) knee has been designed to permit 155 degrees of flexion after total knee arthroplasty (TKA) without compromising wear, polyethylene contact stresses, patellofemoral tracking, or stability. The purpose of this article is to describe the design features of the P.F.C. Sigma RP-F TKA. It is our belief that future primary posterior-stabilized TKA will incorporate many of the design features of the RP-F knee for improved performance.

Arthroplasty, Replacement, Knee↗

Degradation of structurally characterized proteins injected into HeLa cells. Tests of hypotheses.

We have compared sequence and structural features of 35 proteins to their metabolic stabilities in HeLa cells. No relationship was observed between the half-life of an injected protein and its subunit molecular weight, isoelectric point, hydrophobicity, thermostability, surface charge density, or N-terminal residue. Other properties, including susceptibility to oxidation, specific combinations of amino acids, secondary structure composition, and solvent exposed residues, also failed to correlate with protein stability. Although a weak inverse correlation was obtained when stability was compared to asparagine and glutamine content, we conclude that the degradation of an injected protein is unlikely to be related to any single structural parameter. Rather, we hypothesize that it results from an interplay between subcellular location and still poorly defined surface features of the injected proteins.

Amino Acid Sequence↗

Formulation of monocrotophos.

Monocrotophos is an old product that is still very much appreciated as a low-cost and efficient insecticide. Due to its water solubility, formulation recipes are rather simple and no sophisticated equipment is required in a formulation plant. Formulations can easily be manufactured locally where they are going to be used. Formulants and the formulation process do not add much to the total product cost. Monocrotophos is chemically not very stable. A consequence of this property is the marginal shelf life of formulations when stored under tropical conditions. On the other hand, the limited stability of monocrotophos is a most favorable feature in the environment that outweighs the marginal shelf life. By strictly using the "first in-first out" system in stock management, no major problems relating to storage stability will be encountered. An unfavorable characteristic of monocrotophos is its high mammalian toxicity. The objective of future projects will be to develop less toxic formulations.

Drug Stability↗

Catalysis by entropic guidance from enzymes.

Certain features underlying enzymatic catalysis, such as energetic stabilization from binding interactions or proximity and orientation of chemical groups, are evident in the equilibrium-averaged structure of an enzymatic complex determined by crystallography or NMR. Transient features are not apparent from an average structure. Here, we report on a catalytically relevant property of an enzymatic complex revealed by thermal fluctuations from a molecular dynamics study. The conformational fluctuations of the cofactor NADH are altered by binding the enzyme lactate dehydrogenase (LDH) compared to those of free NADH; thermal motions give rise to structures similar to that of the putative transition state. The alteration is stereospecific, in agreement with measured changes in vibrational spectra, and leads to an understanding of the correlation, established some time ago by crystallography and NMR, between the nicotinamide glycosidic bond torsion angle (anti/syn) and the stereospecificity of hydride transfer. These results suggest that one catalytic role of the enzyme is to funnel the population of NADH conformers to the transition state and reduce the entropic barrier to activation. The specific motions in an enzyme complex that might function to enhance transition state formation are described.

Animals↗

A molecular-mechanical model of the microtubule.

Dynamic instability of MTs is thought to be regulated by biochemical transformations within tubulin dimers that are coupled to the hydrolysis of bound GTP. Structural studies of nucleotide-bound tubulin dimers have recently provided a concrete basis for understanding how these transformations may contribute to MT dynamic instability. To analyze these ideas, we have developed a molecular-mechanical model in which structural and biochemical properties of tubulin are used to predict the shape and stability of MTs. From simple and explicit features of tubulin, we define bond energy relationships and explore the impact of their variations on integral MT properties. This modeling provides quantitative predictions about the GTP cap. It specifies important mechanical features underlying MT instability and shows that this property does not require GTP-hydrolysis to alter the strength of tubulin-tubulin bonds. The MT plus end is stabilized by at least two layers of GTP-tubulin subunits, whereas the minus end requires at least one; this and other differences between the ends are explained by asymmetric force balances. Overall, this model provides a new link between the biophysical characteristics of tubulin and the physiological behavior of MTs. It will also be useful in building a more complete description of MT dynamics and mechanics.

Biochemistry↗

Titania and alumina sol-gel-derived microfluidics enzymatic-reactors for peptide mapping: design, characterization, and performance.

The design and characterization of titania-based and alumina-based Poly(dimethylsiloxane) (PDMS) microfluidics enzymatic-reactors along with their analytical features in coupling with MALDI-TOF and ESI-MS were reported. Microfluidics with microchannel and stainless steel tubing (SST) were fabricated using PDMS casting and O(2)-plasma techniques, and were used for the preparation of an enzymatic-reactor. Plasma oxidation for the PDMS microfluidic system enabled the channel wall of the microfluidics to present a layer of silanol (SiOH) groups. These SiOH groups act as anchors onto the microchannel wall linked covalently with the hydroxyl groups of trypsin-encapsulated sol matrix. As a result, the trypsin-encapsulated gel matrix was anchored onto the wall of the microchannel, and the leakage of gel matrix from the microchannel was effectively prevented. A feature of the microfluidic enzymatic-reactors is the feasibility of performing on-line protein analysis by attached SST electrode and replaceable tip. The success of trypsin encapsulation was investigated by AFM imaging, assay of enzymatic activity, CE detection, and MALDI-TOF and ESI-MS analysis. The lab-made devices provide an excellent extent of digestion even at a fast flow rate of 7.0 microL/min, which affords the very short residence time of ca. 2 s. With the present device, the digestion time was significantly shortened compared to conventional tryptic reaction schemes. In addition, the encapsulated trypsin exhibits increased stability even after continuous use. These features are required for high-throughput protein identification.

Aluminum Oxide↗

Amino acid sequences of lysozymes newly purified from invertebrates imply wide distribution of a novel class in the lysozyme family.

Lysozymes were purified from three invertebrates: a marine bivalve, a marine conch, and an earthworm. The purified lysozymes all showed a similar molecular weight of 13 kDa on SDS/PAGE. Their N-terminal sequences up to the 33rd residue determined here were apparently homologous among them; in addition, they had a homology with a partial sequence of a starfish lysozyme which had been reported before. The complete sequence of the bivalve lysozyme was determined by peptide mapping and subsequent sequence analysis. This was composed of 123 amino acids including as many as 14 cysteine residues and did not show a clear homology with the known types of lysozymes. However, the homology search of this protein on the protein or nucleic acid database revealed two homologous proteins. One of them was a gene product, CELF22 A3.6 of C. elegans, which was a functionally unknown protein. The other was an isopeptidase of a medicinal leech, named destabilase. Thus, a new type of lysozyme found in at least four species across the three classes of the invertebrates demonstrates a novel class of protein/lysozyme family in invertebrates. The bivalve lysozyme, first characterized here, showed extremely high protein stability and hen lysozyme-like enzymatic features.

Amino Acid Sequence↗

Meropenem: laboratory and clinical data.

The usefulness of many beta-lactams, including the third-generation cephalosporins, has been threatened in recent years by the dissemination of beta-lactamase-mediated resistance. The carbapenems, meropenem and imipenem/cilastatin, demonstrate ultra-broad spectra of antibacterial activity which encompass the majority of clinically significant Gram-positive and Gram-negative aerobic and anaerobic pathogens. Due to the unique arrangement of the moieties around the carbapenem beta-lactam ring, these agents possess unrivaled stability to almost all serine beta-lactamases, including the mutant extended-spectrum types produced by Enterobacteriaceae which hydrolyze third-generation cephalosporins. Meropenem has a number of additional structural features which confer advantages over imipenem, notably stability to metabolism by dehydropeptidase-I and enhanced activity against Gram-negative organisms, including Pseudomonas aeruginosa. In the treatment of nosocomial respiratory tract infections, meropenem monotherapy has compared favorably with ceftazidime plus aminoglycoside combination regimens or imipenem/cilastatin, producing rates of clinical efficacy of > 75%. Likewise, in patients with intra-abdominal infections, meropenem has proved to be as efficacious as clindamycin plus tobramycin, cefotaxime plus metronidazole and equivalent dosages of imipenem/cilastatin. Meropenem is well tolerated by the central nervous system and, unlike imipenem/cilastatin, is approved for the treatment of meningitis.

Journal Article↗

Activity and thermostability of the small self-splicing group I intron in the pre-tRNA(lle) of the purple bacterium Azoarcus.

The 205-nt group I intron located in the pre-tRNA(lle) from the bacterium Azoarcus sp.BH72 is the smallest self-splicing group I intron identified to date. Comparative sequence analysis has placed this intron and the Anabaena pre-tRNA(Leu) intron into the same subgroup, IC3; we now compare their activity and stability. Unlike the Anabaena intron, the Azoarcus intron has two transitions in the kinetics of the first step of splicing. The faster transition occurs with a larger k(cat)/K(m) than that of the Anabaena or other group I introns, due to a rapid K(cat) (5 min(-1) at 32 degrees C) and a low K(m) for guanosine (17 microM). The excised intron circularizes by releasing a trinucleotide from the 5' end of the intron, another property unlike the Anabaena intron. Although it is smaller in size, the Azoarcus intron retains activity at higher temperatures, higher concentrations of urea, and higher pH than the Anabaena intron. Melting curves show that tertiary structure is disrupted at a lower temperature in the Anabaena intron. Some structural features that may explain the unusual stability of the Azoarcus intron include a G-C rich secondary structure and the presence of two 11-nt motifs, which are known to interact strongly with GAAA loops in group I and group II introns. The disruption of one of these interactions by substituting the Anabaena structural element in fact lowered the thermal stability of the Azoarcus intron. Thus, even superficially similar group I introns from the same structural subgroup can differ significantly in activity and stability.

Base Sequence↗

Thermal, chemical, and enzymatic stability of the cyclotide kalata B1: the importance of the cyclic cystine knot.

The cyclotides constitute a recently discovered family of plant-derived peptides that have the unusual features of a head-to-tail cyclized backbone and a cystine knot core. These features are thought to contribute to their exceptional stability, as qualitatively observed during experiments aimed at sequencing and characterizing early members of the family. However, to date there has been no quantitative study of the thermal, chemical, or enzymatic stability of the cyclotides. In this study, we demonstrate the stability of the prototypic cyclotide kalata B1 to the chaotropic agents 6 M guanidine hydrochloride (GdHCl) and 8 M urea, to temperatures approaching boiling, to acid, and following incubation with a range of proteases, conditions under which most proteins readily unfold. NMR spectroscopy was used to demonstrate the thermal stability, while fluorescence and circular dichroism were used to monitor the chemical stability. Several variants of kalata B1 were also examined, including kalata B2, which has five amino acid substitutions from B1, two acyclic permutants in which the backbone was broken but the cystine knot was retained, and a two-disulfide bond mutant. Together, these allowed determinations of the relative roles of the cystine knot and the circular backbone on the stability of the cyclotides. Addition of a denaturant to kalata B1 or an acyclic permutant did not cause unfolding, but the two-disulfide derivative was less stable, despite having a similar three-dimensional structure. It appears that the cystine knot is more important than the circular backbone in the chemical stability of the cyclotides. Furthermore, the cystine knot of the cyclotides is more stable than those in similar-sized molecules, judging by a comparison with the conotoxin PVIIA. There was no evidence for enzymatic digestion of native kalata B1 as monitored by LC-MS, but the reduced form was susceptible to proteolysis by trypsin, endoproteinase Glu-C, and thermolysin. Fluorescence spectra of kalata B1 in the presence of dithiothreitol, a reducing agent, showed a marked increase in intensity thought to be due to removal of the quenching effect on the Trp residue by the neighboring Cys5-Cys17 disulfide bond. In general, the reduced peptides were significantly more susceptible to chemical or enzymatic breakdown than the oxidized species.

Amino Acid Sequence↗

Human N-acetylgalactosamine-4-sulphatase biosynthesis and maturation in normal, Maroteaux-Lamy and multiple-sulphatase-deficient fibroblasts.

The biosynthesis and maturation of N-acetylgalactosamine-4-sulphatase (4-sulphatase) was studied in normal fibroblasts and in fibroblasts from patients with either mucopolysaccharidosis type VI (MPS VI; Maroteaux-Lamy syndrome) or multiple sulphatase deficiency (MSD). Fibroblasts were incubated in culture medium containing [3H]leucine or [35S]methionine, and radiolabelled 4-sulphatase was isolated by immunoaffinity chromatography using 4-sulphatase-specific monoclonal antibodies. In normal fibroblasts a precursor of 66 kDa, detected intracellularly after 3 h and in NH4Cl-induced secretions, was processed intracellularly, within an additional 3 h, to a polypeptide of 57 kDa composed of disulphide-linked polypeptides of 43 kDa and 8 kDa. All fibroblast lines obtained from MPS VI patients, exhibiting clinical characteristics ranging from no clearly recognized symptoms to the severe classical phenotype, incorporated radioactivity into immune-purified 4-sulphatase at a rate less than 10% of that seen in normal fibroblasts. Maturation of the residual 4-sulphatase showed, variously, features which may be indicative of delayed intracellular transport, decreased intracellular stability, failure of lysosomal targetting or resistance to enzyme processing. Although some features of the residual enzyme synthesis and maturation were consistent with the patient's clinical phenotype, this was infrequent. The maturation of 4-sulphatase in fibroblasts from MSD patients was indistinguishable from that in normal fibroblasts, and the half-life of 4-sulphatase in these fibroblasts, determined after a 24 h pulse and prolonged chase, was only slightly less than that in normal fibroblasts.

Adolescent↗

Cohesion induced by a rotating magnetic field in a granular material.

We report experiments on a magnetic cohesive granular material made of steel spheres in the millimeter range. A magnetic field magnetizes the spheres, so that an interaction force between grains appears. A rotating magnetic field is applied parallel to plane of the quasi-two-dimensional cell containing the spheres so that only the time averaged force between two particles will be considered. Both maximum angle of stability and angles of repose are measured. The maximum angle of stability is found to depend linearly on the interaction force. Another noticeable feature is the lack of dependence of the maximum angle of stability on the initial height of the heap. We show that the angle of repose is less sensitive to the magnetic interaction force than the maximum of stability. At last, we discuss the importance of using a rotating field rather than a constant one. In particular, we report some measurements of both the maximum angle of stability and the angle of repose in constant field, which show a strong dependence of the angles of avalanche on the direction of the field.

Journal Article↗

Don't forget Langmuir-Blodgett films.

Langmuir-Blodgett (LB) films have been investigated, extensively, over the past 70 years. Despite considerable efforts aimed at exploiting their nonlinear optical, piezoelectric, pyroelectric, semiconducting, sensing and barrier properties, problems associated with film quality and stability have hampered their practical development. This feature article highlights one recent advance in the LB area (i.e., the ionic cross-linking of LB films or "gluing"), which has resulted in single bilayer membranes possessing extraordinary quality and stability. The gluing of LB films provides new opportunities for an old technology that has been waning in recent years.

Journal Article↗

Structural stability of adenovirus type 5.

Thermally induced structural changes in adenovirus type 5 (Ad) in the presence of either 2 or 10% sucrose were investigated using a variety of biophysical techniques. In solutions containing 2% sucrose, a highly cooperative transition in the structure of the virus was observed at 45 degrees C as detected by tryptophan fluorescence, derivative UV absorption spectroscopy, circular dichroism (CD), and dynamic and static light scattering. This transition resulted in (at least partial) disassembly of the virus and a concomitant increase in the accessibility of the viral DNA to the fluorescent dye, TOTO-1. Capsid disassembly was verified by transmission electron microscopy, which showed ruptured icosahedral vertices near 45 degrees C followed by complete capsid disassembly at higher temperatures. SDS-PAGE of thermally treated Ad suggests that the penton base (protein III) and protein IIIa (located in the peripentonal region) are significantly more labile than other capsid proteins and may be the initial instigators in capsid disassembly. Additional discrete structural transitions were observed in viral proteins using the aforementioned spectroscopic techniques. Thermally induced rearrangements of the condensed DNA at higher temperatures were also detected by the appearance of "psi"-like features in the CD spectra as well as a dramatic decrease in accessibility of DNA to TOTO-1. These transitions corresponded to discrete endothermic events that are also detected by differential scanning calorimetry. By increasing the concentration of sucrose to 10%, secondary and tertiary structural features of adenoviral proteins were significantly stabilized, although loss of quaternary structure at 45 degrees C was still observed.

Adenoviridae↗

Stability of cell-penetrating peptide-morpholino oligomer conjugates in human serum and in cells.

Cell penetrating peptides (CPPs) have been shown to enhance the cellular uptake of antisense oligonucleotides (AOs). However, the effectiveness of the CPPs for cytoplasmic or nuclear delivery of therapeutic AOs must take into account the possible entrapment of the CPP-AO conjugates in endosomes/lysosomes and the overall stability of the CPP-AO conjugates to enzymes. This includes the stabilities of the CPPs and AOs themselves as well as the linkage between them. In this study, we investigated the effects of several structural features of arginine-rich CPPs on the metabolic stability of CPP conjugated to phosphorodiamidate morpholino oligomers (PMOs) in human serum and in cells. Those structural features include amino acid configurations (d or l), incorporation of non-alpha-amino acids, peptide sequences, and types of linkages between CPPs and PMOs. Using matrix-assisted laser desorption ionization time-of-flight mass spectrometry, we found that the stability of the CPP portion was varied although the PMO portion of the conjugate was completely stable both in cells and in human serum. d-Configuration CPPs were completely stable, while l-CPPs were degraded in both serum and HeLa cells. Insertions of 6-aminohexanoic acid residues (X) into an R8 peptide increased the corresponding CPP's serum stability with the degree of stability being dependent upon the positions of X. However, X-containing CPPs were degraded rapidly intracellularly. Insertions of beta-alanines (B) into the R8 peptide increased its serum stability and intracellular stability. An amide or a maleimide linkage was stable in both serum and cells; however, an unhindered disulfide linkage was not stable in either. By using fluorescent microscopy, flow cytometry, and an antisense splice correction assay, the cellular uptakes of an X-containing conjugate and its fragments were compared to their antisense activities. We found that a large fraction of the conjugate was trapped within vesicles and the degraded fragments cannot escape from the vesicles. This study indicates that the incorporation of non-alpha-amino acids into l-CPPs can increase the metabolic stability of CPP-PMOs without using costly d-CPPs. However, the position and type of non-alpha-amino acids affect the degree of stability extracellularly and intracellularly. In addition, this study reveals that the degradation of an X-containing CPP-PMO conjugate is a more rapid process than degradation of a B-containing conjugate. Last, the endosomal/lysosomal trapping limits the effectiveness of a CPP-PMO conjugate, and the stability of the CPP is one of the factors affecting the ability of the conjugate to escape the endosomes/lysosomes.

Cell Membrane↗

The evolution of protamine P1 genes in dasyurid marsupials.

We report the complete DNA sequences of the protamine P1 gene and flanking regions for 13 species of the marsupial family Dasyuridae. The structure of the protamine locus is conserved in dasyurids and consists of two exons (of lengths 142-151 and 47 bp) separated by an intron (208-240 bp). A key feature of the dasyurid intron is a 38-40 bp duplication found in all species examined to date. This duplication apparently predates the radiation of modern dasyurid lineages and may be homologous to a similar feature in the marsupial mole (Notoryctes). Sequences from a species of Planigale demonstrate that this genus is unique among marsupials in possessing cysteine residues in its protamine P1 molecules. Cysteines may provide enhanced chemical stability for condensed sperm nuclei, a physiological feature that would converge on the common eutherian pattern. Phylogenetic analysis of the protamine genes yields a tree that is largely congruent with previous molecular systematic studies in two areas: (1) There are three main dasyurid lineages corresponding to the Sminthopsinae. Dasyurinae, and Phascogalinae; (2) Dasyurinae and Phascogalinae are sister groups. This study is the first estimate of dasyurid relationships based on a nuclear DNA sequence.

Amino Acid Sequence↗