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A composite micromechanical model for connective tissues: Part I--Theory.

A micromechanical model has been developed to study and predict the mechanical behavior of fibrous soft tissues. The model uses the theorems of least work and minimum potential energy to predict upper and lower bounds on material behavior based on the structure and properties of tissue components. The basic model consists of a composite of crimped collagen fibers embedded in an elastic glycosaminoglycan matrix. Upper and lower bound aggregation rules predict composite material behavior under the assumptions of uniform strain and uniform stress, respectively. Input parameters consist of the component material properties and the geometric configuration of the fibers. The model may be applied to a variety of connective tissue structures and is valuable in giving insight into material behavior and the nature of interactions between tissue components in various structures. Application of the model to rat tail tendon and cat knee joint capsule is described in a companion paper [2].

Biomechanical Phenomena

Stabilization of the nuclear matrix by disulfide bridges: identification of matrix polypeptides that form disulfides.

The molecular structure of the nuclear matrix is still poorly understood. We have tried to assess which proteins are important structural elements by examining the process of stabilization of the nuclear matrix by sodium tetrathionate. Sodium tetrathionate stabilizes the nuclear matrix by oxidizing sulfhydryl groups to disulfides. We show that tetrathionate-stabilized matrices are disassembled in buffers containing SDS, indicating that the stabilized nuclear matrix is not a continuous network of cross-linked proteins. Using monobromobimane, a thiol-specific fluorescent reagent, we show that many protein thiols in the stabilized matrix are oxidized. By chromatography on activated thiol-Sepharose we estimated that about 50% of the matrix proteins had oxidized sulfhydryl groups. The protein composition of the material bound to activated thiol-Sepharose was similar to that of the not-bound material. A few proteins are highly enriched in the fraction that was bound to the column. This indicates that many matrix protein species are partially oxidized and that some proteins are completely oxidized. The oxidized protein thiols are found in relatively large complexes as determined by SDS gel-electrophoresis under nonreducing conditions. These results are interpreted in terms of protein-protein interactions in the matrix. The possible role of thiols and disulfides in the in vivo organization of the nucleus is discussed.

Animals

Predicting food taste with bound-driven optimization.

The prediction of sensory attributes from ingredient-level formulations is an emerging challenge at the intersection of food science and artificial intelligence. We address the fundamental question of whether the taste of a food can be predicted from its ingredients by treating recipes as composite materials. We apply Hashin-Shtrikman (HS) and Reuss-Voigt (RV) bounds, techniques originally developed for elastic moduli, as a null-hypothesis additive baseline for five taste dimensions (sweetness, sourness, bitterness, umami, saltiness) on a curated dataset of 70 recipes decomposed into 115 distinct ingredients scored against a library of 209 ingredient-level taste references with trained-panel ground truth. This baseline systematically under-predicts perceived taste: 77% of actual taste values exceeded the HS upper bound, with the exceedance rate ranging from 26% (bitterness) to 97% (saltiness). We traced this gap to specific processing chemistry (Maillard reactions, caramelization, evaporative concentration, protein hydrolysis, and nucleotide synergy) and introduced a hybrid model that augments the HS baseline with eight chemistry-proxy features encoding these mechanisms. Our results show that our interpretable hybrid model eliminates the systematic bias and reduces mean absolute error by 27%-62% for sweetness, sourness, umami, and saltiness while using only 10 interpretable features, achieving performance comparable to a black-box Lasso regression on 115 per-ingredient features. We further demonstrate constrained inverse design via Differential Evolution, recovering ingredient formulations that match target taste profiles subject to compositional bounds. Our work demonstrates how key chemical processes during food preparation can inform and augment physics-based and machine learning models, providing a quantitative fingerprint of processing chemistry's contribution to taste perception and paving the way for model-driven food formulation with targeted sensory characteristics.

Composite material bounds

A composite micromechanical model for connective tissues: Part II--Application to rat tail tendon and joint capsule.

A micromechanical model of fibrous soft tissue has been developed which predicts upper and lower bounds on mechanical properties based on the structure and properties of tissue components by Ault and Hoffman [3, 4]. In this paper, two types of biological tissue are modeled and the results compared to experimental test data. The highly organized structure of rat tail tendon is modeled using the upper bound aggregation rule which predicts uniform strain behavior in the composite material. This model fits the experimental data and results in a correlation coefficient of 0.98. Applied to cat knee joint capsule, the lower bound aggregation rule of the model correlates with the data and predicts uniform stress within this more loosely organized tissue structure. These studies show that the nature of the interactions between the components in tissue differs depending upon its structure and that the biomechanical model is capable of analyzing such differences in structure.

Animals

Chemical analysis of stalk components of Dictostelium discoideum.

Structural components of the stalks of mature fruiting bodies of Dictyostelium discoideum have been isolated and characterized after solubilizing non-structural components with urea and sodium dodecyl sulfate. The urea/sodium dodecyl sulfate-insoluble stalks are composed of about 52% cellulose, 15% protein and 3% of a non-cellulosic heteropolymer in a covalently bound matrix. Non-covalently bound fatty acid containing material was also found. The composition and structural interrelationships of these components are essentially identical to that of the urea/sodium dodecyl sulfate-insoluble surface sheath which is produced earlier in development before culmination. These results suggest that the same components are involved in making structural elements which differ substantially in their functional role in the developmental sequence as well as in their spatial and temporal localization and morphological appearance.

Amino Acids

Composition of diesel exhaust with particular reference to particle bound organics including formation of artifacts.

For particulate emissions, standards were established by the US EPA in February 1980. Regulations limiting particulates from new light duty diesel vehicles are valid by model year 1982. The corresponding standards on a pure mass basis do not take into account any chemical character of the diesel particulate matter. Our investigation of the material composition shows that diesel particulates consist mainly of soot (up to 80% by weight) and adsorptively bound organics including polycyclic aromatic hydrocarbons (PAH). The qualitative and quantitative nature of hydrocarbon compounds associated with the particulates is dependent not only on the combustion parameters of the engine but also to an important degree on the sampling conditions when the particulates are collected (dilution ratio, temperature, filter material, sampling time etc.). Various methods for the analyses of PAH and their oxy- and nitro-derivatives are described including sampling, extraction, fractionation and chemical analysis. Quantitative comparison of PAH, nitro-PAH and oxy-PAH from different engines are given. For assessing mutagenicity of particulate matter, short-term biological tests are widely used. These biological tests often need a great amount of particulate matter requiring prolonged filter sampling times. Since it is well known that facile PAH oxidation can take place under the conditions used for sampling and analysis, the question rises if these PAH-derivates found in particle extracts partly or totally are produced during sampling (artifacts). Various results concerning nitro- and oxy-PAH are presented characterizing artifact formation as a minor problem under the conditions of the Federal Test Procedure. But results show that under other sampling conditions, e.g. electrostatic precipitation, higher NO2-concentrations and longer sampling times, artifact formation can become a bigger problem. The more stringent particulate standard of 0.2 g/mi for model years 1986 and 1987 respectively requires particulate trap technology. Preliminary investigations of the efficiency of ceramic filters used reveal that the reduction of the adsorptively bound organics is lower than the decrease of the solid carbonaceous fractions.

Aldehydes

Large scale isoelectric focusing analysis of the non-histone proteins of the nucleus: isolation of components with alkaline isoelectric points.

After large scale isoelectric focusing of rat liver non-histone protein in polyacrylamide gel, pH range 4--8.6, the only protein material found outside the gradient was present in the cathode solution (20 mM NaOH). This was low mol. wt protein material (approximately 10,000) with an acidic amino acid composition. It bound 5--6 times its own weight of basic ampholine carrier ampholytes to give a complex with a pI of 8.82. This could be dissociated by dialysis against 1 M NaCl.

Animals

Metabolic changes in elicitor-treated bean cells. Enzymic responses associated with rapid changes in cell wall components.

Treatment of cell suspension cultures of bean (Phaseolus vulgaris c.v. Immuna) with an elicitor preparation heat-released from the cell walls of the phytopathogenic fungus Colletotrichum lindemuthianum resulted in rapid changes in the composition of the bean cell walls. These consisted of (a) increases in phenolic material bound to the cellulosic and hemicellulosic fractions of the wall, (b) loss of material (mainly glucose) from the hemicellulosic fraction and (c) an increase in wall-associated hydroxyproline. The increases in wall-bound phenolics were preceded by (a) rapid decreases in the intracellular levels of free hydroxycinnamic acids and (b) transient increases in the extractable activities of L-phenylalanine ammonia-lyase and cinnamic acid 4-hydroxylase. 4-Hydroxycinnamic acid 3-hydroxylase activity was present at a high level in control cultures and was not induced by elicitor. Changes in the levels of cytochrome P-450, as determined by dot blot assays utilising an anti-(P-450) monoclonal antibody, paralleled the changes in cinnamic acid 4-hydroxylase activity. The accumulation of cell wall hydroxyproline was associated with rapid transient increases in the extractable activities of proline 2-oxoglutarate dioxygenase and a protein arabinosyl transferase. An hydroxyproline-rich acceptor protein of Mr 42 500 was the major protein to incorporate [3H]arabinose following elicitation of the bean cells, and the kinetics of the extent of labelling of this protein paralleled the accumulation of hydroxyproline protein in the endomembrane system. The above metabolic changes associated with cell wall components followed rapid kinetics similar to those involved in the formation of the phytoalexin kievitone in the elicited cultures [Robbins, M. P. et al. (1985) Eur. J. Biochem. 148, 563-569]. It is therefore concluded that increased 5-hydroxy-substituted isoflavonoid biosynthesis, wall-bound phenolic synthesis and synthesis of arabinosylated hydroxyproline-rich protein are all early events which are closely linked to the initial interaction between plant cell and fungal elicitor.

Ammonia-Lyases

The structure and ultrastructure of the rostral cartilage in the spiny eel, Macrognathus siamensis (Teleostei: Mastacembeloidei).

An elastic, cell-rich cartilage provides flexible support to the highly mobile, rostral tentacle of the mastacembelid, Macrognathus siamensis. Active movement of the tentacle is effected by skeletal muscles, the muscular bellies of which are located outside the organ. The tentacle returns to its original shape by elastic recoil. The cartilage resembles plant supporting tissue and the cartilages of certain invertebrates. It is surrounded by a thick perichondrium and articulates at a synovial joint with the supraethmoid. The chondrocytes are large and shrunken within lacunae. They contain glycogen and cytoplasmic stores of RNA. The matrix is reduced to thin seams between adjacent cells, and stains strongly with alcian blue and a variety of elastic stains. Parts of the matrix are trilaminar, and such an appearance recalls the distinction between the primary cell walls of adjacent plant cells and the intervening middle lamella. The perichondrium consists of an alternating sequence of cells, circularly arranged collagen fibres and a foamy, amorphous material of unknown composition. Deep to the perichondrium, the chondrocytes are packed with intermediate filaments. Membrane-bound organelles are not prominent, though mitochondria are located at the periphery of the cell. The ultrastructural similarities between these chondrocytes and those of hyaline-cell cartilage (chondroid) support the contention that Schaffer's concept of a rigid distinction between 'true' cartilage and 'chondroid' is no longer tenable. The matrix is devoid of collagen and is most distinctive. In the immediate vicinity of the cells it consists of matrix granules and matrix fibers but, where a trilaminar appearance is easily discernible, there is a central zone that consists of large masses of amorphous material that is presumed to contain elastin. Such amorphous material has not previously been seen in any teleostean elastic tissue. By contrast, elastic system fibres, readily demonstrable elsewhere in teleosts, are conspicuously absent.

Animals

The role of surface polysaccharide in determining the resistance of Serratia marcescens to serum killing.

Two O14:H12 strains of Serratia marcescens with different sensitivities to killing by normal pooled human serum were investigated. Complement binding, studied by measuring hydrophobicity and using rocket immunoelectrophoresis with anti-human C3, showed the sensitive cells (S1220) rapidly bound and fixed complement whereas the resistant cells (4444-60) bound less C3b. The strains had identical membrane protein composition. Crossed immunoelectrophoresis suggested that in S1220 cells the polysaccharide material including LPS was less antigenic and present in smaller amounts than in 4444-60 cells. This was confirmed by examining extracted polysaccharide material chemically and by SDS-PAGE. The resistant strain had 33% more phenol-extractable polysaccharide material than the sensitive strain, possibly comprising LPS with longer O antigen chain lengths, or a microcapsule of O antigen polysaccharide. Extra polysaccharide material on the surface of the resistant strain prevents complement components binding and reaching the hydrophobic membrane where lytic lesions occur.

Antigens, Bacterial

Effect of silicone oil on protein adsorption to hydroxyapatite in vitro and on pellicle formation in vivo.

Silicone oil has been introduced in a dentifrice for smokers because of its effect as a polishing agent. Silicone oils are hydrophobic in character and have low surface tensions and good wetting properties. Due to the low surface tension, silicone oils may spread readily on solid surfaces and cover them with a thin, water-repellent film. Introduced via dentifrices silicone oil may thus well be able to adsorb to enamel surfaces and to interfere with surface characteristics such as protein adsorption. The aim of the present study was to examine the effect of silicone oil on protein adsorption to hydroxyapatite (HA) in vitro and on pellicle formation in vivo. The effect on protein adsorption to HA in vitro was studied by adsorption of albumin to either untreated or silicone oil treated HA powders. Ion exchange chromatography was also used with either untreated or silicone oil treated HA as bed materials. The effect on pellicle formation in vivo was studied using enamel fragments carried in the mouth to acquire pellicle material. The chemical composition of the acquired pellicle was studied by collection and chemical analysis of pellicle material formed on enamel surfaces in vivo. The study showed that silicone oil treated HA took up less protein and that the adsorbed protein was bound to hydroxyapatite by a different mechanism as compared to untreated controls. The results indicated that hydrophobic interactions could be involved in binding of proteins to silicone oil treated hydroxyapatite. Silicone oil treated enamel fragments carried in the mouth showed a slower rate of pellicle formation as compared to untreated fragments. The amino acid composition of the acquired pellicle collected in vivo from silicone oil treated enamel surfaces was also different from pellicle material collected from untreated enamel.

Adsorption

Isolation of nascent high-density lipoprotein from rat liver perfusates by immunoaffinity chromatography: effects of oleic acid infusion.

Immunoaffinity chromatography on a column of rabbit IgG anti-rat apolipoprotein (apo) A-I covalently bonded to agarose was used to isolate nascent high-density lipoprotein (nHDL) from recirculated perfusates of rat livers. After passage through the affinity column, the bound material was eluted with sodium thiocyanate and analyzed for apolipoproteins and lipids. The protein content was 52% and the lipid composition was 37% triglyceride, 40% phospholipid, and 23% cholesterol. Apolipoproteins E and A-I each comprised approximately one third of the total, and very little apo B was detectable as judged by SDS-PAGE analysis. The affinity-isolated particles were therefore similar in composition to the major apo A-I:apo E-rich subfraction of nHDL isolated by ultracentrifugation in earlier work. It is concluded that the apo E in this class of nHDL (containing both apo E and apo A-I) is present in the secreted particle and is not a consequence of a loss of apo E from very-low-density lipoprotein (VLDL) during ultracentrifugation. The high triglyceride content in the virtual absence of apo B confirms and extends previous analyses and reinforces the conclusion that nHDL particles are enriched in triglyceride compared to plasma HDL. The inclusion of 4% albumin in the perfusion medium did not significantly change the total triglyceride output of 115 micrograms/g liver/h, but it decreased the triglyceride output isolated by anti-apo A-I affinity chromatography from 3.2 to 0.48 micrograms/g liver/h. The addition of oleic acid complexed to albumin increased the total triglyceride output by 70% and that associated with the immunoaffinity column increased from 0.48 to 2.7 micrograms/g liver/h.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Remobilization of toxic heavy metals adsorbed to bacterial wall-clay composites.

Significant quantities of Ag(I), Cu(II), and Cr(III) were bound to isolated Bacillus subtilis 168 walls, Escherichia coli K-12 envelopes, kaolinite and smectite clays, and the corresponding organic material-clay aggregates (1:1, wt/wt). These sorbed metals were leached with HNO3, Ca(NO3)2, EDTA, fulvic acid, and lysozyme at several concentrations over 48 h at room temperature. The remobilization of the sorbed metals depended on the physical properties of the organic and clay surfaces and on the character and concentration of the leaching agents. In general, the order of remobilization of metals was Cr much less than Ag less than Cu. Cr was very stable in the wall, clay, and composite systems; pH 3.0, 500 microM EDTA, 120-ppm [mg liter-1] fulvic acid, and 160-ppm Ca remobilized less than 32% (wt/wt) of sorbed Cr. Ag (45 to 87%) and Cu (up to 100%) were readily removed by these agents. Although each leaching agent was effective at mobilizing certain metals, elevated Ca or acidic pH produced the greatest overall mobility. The organic chelators were less effective. Lysozyme digestion of Bacillus walls remobilized Cu from walls and Cu-wall-kaolinite composites, but Ag, Cr, and smectite partially inhibited enzyme activity, and the metals remained insoluble. The extent of metal remobilization was not always dependent on increasing concentrations of leaching agents; for example, Ag mobility decreased with some clays and some composites treated with high fulvic acid, EDTA, and lysozyme concentrations. Sometimes the organic material-clay composites reacted in a manner distinctly different from that of their individual counterparts; e.g., 25% less Cu was remobilized from wall- and envelope-smectite composites than from walls, envelopes, or smectite individually in 500 microM EDTA. Alternatively, treatment with 160-ppm Ca removed 1.5 to 10 times more Ag from envelope-kaolinite composites than from the individual components. The particle size of the deposited metal may account for some of the stability changes; those metals that formed large, compact aggregates (Cr and Ag) as seen by transmission electron microscopy were less likely to be remobilized. In summary, it is apparent that remobilization of toxic heavy metals in sediments, soils, and the vadose zone is a complicated issue. Predictions based on single inorganic or organic component systems are too simplistic.

Adsorption

Amino acid analysis of proteins and tightly bound phosphopeptides in calf thymus DNA: indications of a novel regulatory mechanism.

Purification of DNA prepared from calf thymus nuclei, which were isolated at temperatures of -10 degrees to -15 degrees C, termed N-DNA, reduced the protein content from 17% to 0.7%. Amino acid analysis of the crude N-DNA revealed that the protein material removed in this process contained lys and phe, whereas in purified N-DNA neither of these amino acids was present. The crude N-DNA had a lower total Pser molar ratio (5.9) than that of the purified N-DNA (22.6). The purification also effected a large decrease of molecular weight, from 170 to 15 million daltons, roughly corresponding to the size of replicons. The amino acid composition of the N-DNA was compared with those of the two PP fractions, a) the PPs released into the dialysates, and b) the core material bound in the DNA, probably covalently, after reaction of N-DNA with EDTA and subsequent dialysis. N-DNA had an intermediate total Pser molar ratio between that of a) (35.3) and b) (11.9). Analysis of a standard preparation of DNA (S-DNA), which involved initial rupture of nuclei in the presence of cytoplasmic components, gave elevated values of asp and glu, a total Pser average value of 8.7 before, and 9.0, after the purification, and again phe and lys in the crude DNA.

Amino Acids

[A carbohydrate component of alpha1-acid glycoprotein from normal and abortive blood].

alpha 1-Acid glycoprotein (AGP) samples, viz., normal AGP (nAGP) and embryonic AGP (eAGP), were isolated from plasma of normal donor blood and abortive human blood, respectively. The materials possess similar amino acid composition and immunochemical properties. However, structural patterns of their sugar moieties differ substantially. nAGP contains only N-glycosidically bound sugar chains, whereas on eAGP some sugar chains are linked through O-glycosidic bonds. Disaccharide GlcNAc beta 1----3Gal was identified in considerable amounts in HF-solvolysate of eAGP but only as traces in nAGP.

Amino Acids

Lipid and cell wall changes in an inositol-requiring mutant of Neurospora crassa.

An inositol deficiency in the inositol-requiring (inl) mutant of Neurospora crassa led to changes in the composition of the inositol-containing lipids and the cell wall. On deficient levels of inositol, phosphatidyl inositol decreased by 23-fold, di(inositolphosphoryl) ceramide decreased by 4-fold, and monoinositolphosphoryl ceramide increased slightly. The inositol deficiency also led to an aberrant hyphal morphology and changes in both the amount of cell wall and the amino sugar content of the cell wall. The glucosamine content of the cell wall decreased by 50%, the galactosamine increased by 50%, but no significant changes were found in the content of the cell wall amino sugar precursors, or in the amino acid, glucose, or total hexose content of the cell wall. Inositol-containing compounds were found associated with purified cell wall material. These compounds were bound tightly to the cell wall but could be removed by treatment with alkali, a treatment which disrupts the cell wall integrity. Possible mechanisms of how changes in lipid composition can affect cell wall biosynthesis are discussed.

Cell Wall

Characterization of lamina-bound chromatin in the nuclear shell isolated from HeLa cells.

The structure and the polypeptide composition of the nuclear shell isolated from interphase HeLa cells have been investigated and compared to those of the intranuclear material. The isolated nuclear shell contains chromatin superstructures (28-32 nm thick fibres) made of tightly packed nucleosomes that resist low ionic strength conditions and that are associated with the three nuclear lamins. Chromatin in the nuclear shell exhibits very simple chemical composition. Especially, non-histone proteins are lacking. The results presented here rule out the possibility that the nuclear shell results from contamination of lamina by intranuclear elements. They suggest that the lamins are directly involved in the specific properties and in the organization of chromatin in the nuclear shell.

Cell Fractionation

[Bonded porcelain laminates--an alternative to anterior porcelain crowns].

The trend towards esthetic dentistry, which originates in the USA, has added new dimensions to the treatment of anterior teeth. One of it is the porcelain laminate technique. Using composites, extremely thin, manually prepared porcelain laminates with a thickness of 0.3 to 0.5 mm are bounded to the labial aspect of front teeth or laterals. On account of the low laminate thickness very little enamel preparation is needed. The residual enamel mass and the translucency of the ceramic material produce a natural appearance and optimal cosmetic results. This technique enables the dentist to restore the shape and color of esthetically compromised teeth.

Composite Resins