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RAPD profile variation amongst provenances of neem.

Neem, described as a tree for solving global problems, is an evergreen, long-lived, multipurpose tree of the tropics with a wide distribution range in India. It is believed to be highly cross-pollinated. Inter-provenance variations have been reported in neem in case of morphological and physiological characters. Yet no reports about the genetic determinism for these variations are available to our knowledge. In order to have an idea about the extent and/or nature of genetic (DNA) variation in neem, the powerful RAPD technique has been employed. RAPD profiles of 34 accessions/provenances of neem were generated with 200 decamer random primers, of which the data from the 49 primers, that resulted in reproducible amplification products, were considered for analysis. Based on the presence/absence of bands, a similarity matrix was computed. Dendrogram was constructed by UPGMA method based on the pairwise similarities amongst the RAPD profiles. The similarities in RAPD profiles amongst the different DNAs was more than that expected due to the cross-pollinated nature of the tree and furthermore, these more-than-expected similarities were not due to random chance. These results suggest that neem may have a narrow genetic base.

DNA Primers↗

The yeast mitochondrial intermembrane space: purification and analysis of two distinct fractions.

We have developed a protocol for the sequential release of the intermembrane space (IMS) content of Saccharomyces cerevisiae mitochondria. Two distinct fractions were obtained: a soluble IMS with cytochrome b2 as key marker and a salt-extractable IMS with cytochrome c as key marker. The identity of several proteins was determined by amino-terminal amino acid sequencing. The IMS fractions were devoid of contaminations from cytosol and mitochondrial outer and inner membranes. By subtraction analysis, the protein profiles of soluble and salt-extractable IMS fractions were depleted of contaminating bands derived from matrix proteins. The fractionation method will provide the basis for the further analysis of IMS proteins and characterization of their functions in bioenergetics, mitochondrial biogenesis, and regulatory processes.

Cell Fractionation↗

[Electron microscopic investigation of connective tissue in vestibular ganglion and nerve in morbus Menière].

Vestibular ganglia of 17 patients with Menière's disease, obtained by transtemporal or translabyrinthine neurectomy, were studied by electron microscopy. Three ganglia removed because of other disease and four ganglia of normal ears taken post mortem served as controls. The neuronal fibrous tissue of Menière cases showed without exception pathologic changes of various extent. The amount of collagen was increased, whereby different fiber diameters were observed as well as changes in the periodicity of cross banding. Signs of continuing collagen formation were found: active fibroblasts and an increased number of isolated Schwann cells without axons, showing deerhorn-like ramifications which enveloped collagen bundles. The blood vessels were frequently surrounded by multiple basal membranes and broad bands of homogenous matrix. The pericytes were either necrotic or nonexistent. The endothelial cell cytoplasma was usually not in an active state. Sometimes it seemed to be autolytic. The pinocytotic activity was strikingly diminished. These qualitative changes of the interstitial tissue might point to a local pathologic event in the region of the vestibular nerve and ganglion.

Connective Tissue↗

Magnetic resonance imaging assessment of brain maturation in preterm neonates with punctate white matter lesions.

INTRODUCTION: Early white matter (WM) injury affects brain maturation in preterm infants as revealed by diffusion tensor imaging and volumetric magnetic resonance (MR) imaging at term postmenstrual age (PMA). The aim of the study was to assess quantitatively brain maturation in preterm infants with and without milder forms of WM damage (punctate WM lesions, PWML) using conventional MRI. METHODS: Brain development was quantitatively assessed using a previously validated scoring system (total maturation score, TMS) which utilizes four parameters (progressive myelination and cortical infolding, progressive involution of glial cell migration bands and germinal matrix tissue). PWML were defined as foci of increased signal on T1-weighted images and decreased signal on T2-weighted images with no evidence of cystic degeneration. A group of 22 preterm infants with PWML at term PMA (PWML group) were compared with 22 matched controls with a normal MR appearance. RESULTS: The two groups were comparable concerning gestational age, birth weight and PMA. TMS was significantly lower in the PWML group than in the control group (mean TMS 12.44 +/- 2.31 vs 14.00 +/- 1.44; P = 0.011). Myelination (mean 2.76 +/- 0.42 PWML group vs 3.32 +/- 0.55 control group, P = 0.003) and cortical folding (3.64 +/- 0.79 vs 4.09 +/- 0.43, P = 0.027) appeared to be significantly delayed in babies with PWML. CONCLUSION: Conventional MRI appears able to quantify morphological changes in brain maturation of preterm babies with PWML; delayed myelination and reduced cortical infolding seem to be the most significant aspects.

Brain↗

Genetic diversity among Borrelia burgdorferi isolates: more than three genospecies?

In order to assess the genetic relationships among isolates of Borrelia burgdorferi, we have determined chromosomal DNA restriction fragment length polymorphisms (RFLP) of 27 isolates. Using five restriction enzymes and seven DNA probes, we recorded 246 different hybridizing bands. The resulting matrix was subjected to factorial correspondence analysis and to phylogenetic analysis using a parsimony program. These analyses demonstrate two closely related genospecies and three individual strains. RFLP variations among these three strains are much more pronounced than among the members of the other two genospecies. This may indicate the existence of more than three genospecies. Plasmid analysis did not contribute significantly to the genotypic differentiation.

Borrelia burgdorferi Group↗

Pilot study of the effects of three bonding systems on the torsional resistance of a titanium-reinforced composite core.

STATEMENT OF PROBLEM: There are no studies analyzing the effects of a bonding agent on torsional resistance for a titanium-reinforced core material bonded with a multistep bonding system. PURPOSE: This pilot study investigated the effects of multistep bonding agents on torsional resistance of a titanium-reinforced composite core material (Ti-Core core material). MATERIAL AND METHODS: A total of 40 recently extracted human molars with their crowns removed were placed into acrylic resin blocks and divided into 4 groups of 10 specimens. Only multistep bonding systems were used (AllBond-2, ScotchBond Multi- PURPOSE: Tenure A&B, and the control without bonding agent). Specimens were prepared according to manufacturer's instructions. The control group was etched with a 37.5% phosphate acid. All cores were fabricated with Ti-Core titanium core material with No. 1 hard copper bands as the matrix. Ti-Core core material was applied and allowed to cure for 1 hour, then the cores were placed in 100% humidity (water) for 1 week. Test specimens were placed in a special jig and a clockwise torsional force applied. Torsional force was measured on a Lebow 1102-200 torque transducer and recorded on a Hewlett Packard 7015B X-Y recorder in inch x ounce then converted into Newton-meter. One-way analysis of variance (ANOVA) was used to test for statistical significance (P <.05). RESULTS: Torsional values (Newton-meter) were AllBond-2, 0.1980, ScotchBond Multi- PURPOSE: 0.1890, Tenure A&B, 0.2142; the control group recorded a value of zero. Statistical analysis revealed that bonding systems did not differ from one another. CONCLUSION: Use of a bonding agent increased the core's resistance to torsional forces. Lack of a bonding agent dramatically reduced the resistance to torque.

Analysis of Variance↗

Fine structure of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 (Monogenea: Ancyrocephalidae).

A study of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 was carried out using light and electron microscopy. The anterior adhesive apparatus or head organs in B. gussevi comprise 6 circular openings or apertures in the antero-lateral region, associated pits lined with specialized microvillous tegument that differ from the general body tegument, a bundle of ducts, and uninucleate gland cells located lateral to the pharynx. The uninucleate glands of the anterior adhesive apparatus (head organs) comprise 2 types of cells, one kind of cell producing rod-like bodies (S1) and the other oval bodies (S2). The S1 bodies are filled with numerous, less electron-dense vesicles in an electron-dense matrix, while S2 bodies have no vesicles but contain a more homogeneous electron-dense matrix. Interlinking band-like structures were observed between S1 bodies. Similar band-like structures were found between S2 bodies. The formation of S1 bodies was followed by transmission electron microscopy. However, the formation of S2 bodies was unclear and could not be resolved. Uniciliated structures were also observed around the openings of the anterior adhesive apparatus. Each uniciliated structure is usually associated with an opening of a gland cell producing granular, electron-dense, secretory bodies, which differ from the secretions produced by the lateral gland cells of the anterior adhesive apparatus.

Animal Structures↗

Remarkable metal-rich ternary chalcogenides Sc14M3Te8 (M = Ru, Os).

In this novel motif, scandium atoms define infinite parallel chains of alternate trans-face-sharing cubes and pairs of square antiprisms in which each polyhedron is also centered by an M atom (M = Ru, Os). These chains are further linked into a three-dimensional structure by Sc(Te2Te4/2) octahedra. Physical property measurements show Sc14Ru3Te8 to be metallic and Pauli-paramagnetic, consistent with the results of extended Hückel band structure calculations. Matrix effects are evident in the dimensions within the chains. The major interactions are Sc-M and Sc-Te.

Journal Article↗

Torsional resistance of crowns cemented to composite cores involving three stainless steel endodontic post designs.

STATEMENT OF PROBLEM: There are no studies analyzing the effects of a crown in relationship to torsional resistance for a titanium-reinforced core material supported by a post. PURPOSE: This study investigated the effects of post design on the torsional resistance of a crown supported by a titanium-reinforced composite core material (Ti-Core) and 3 endodontic posts (AccessPost, Flexi-Post, and ParaPost). MATERIAL AND METHOD: Three groups of 10 specimens per group were studied. A total of 30 recently extracted human single-rooted (incisors) teeth with their crowns removed were used. Post holes preparations were created according to manufacturer's instructions. All posts were cemented using Flexi-Flow with titanium cement. Scotchbond Multi-Purpose bonding system was used as the bonding system for all groups. All cores were fabricated with Ti-Core with titanium core material with hard copper bands as the matrix. The Ti-Core core material was allowed to set for 1 hour. Metal crowns were fabricated and cemented with zinc phosphate cement. Specimens were placed in a special jig and a clockwise torsional force was applied. Torsional force was measured on a Lebow 1102-200 torque transducer and recorded on a Hewlett Packard 7015B X-Y recorder in inch x ounce. A 1-way analysis of variance (ANOVA) was used to test for statistical significance (P < .001). RESULTS: The torsional values (inch x ounce) were AccessPost 77.2 +/- 30.3, Flexi-Post 162.2 +/- 51.2, and ParaPost 60.9 +/- 28.4. SNK and Scheffé comparison tests revealed that the Flexi-Post group had significantly greater resistance to torque loading than the AccessPost and ParaPost groups, which were statistically similar to each other. CONCLUSION: Post design has an effect on torsional resistance of a crown supported by titanium-reinforced core build-up and post. The threaded split-shank design of the Flexi-Post dowel offers significantly greater resistance to torsional loading than the passive posts studied.

Analysis of Variance↗

Reversible unfolding of fibronectin type III and immunoglobulin domains provides the structural basis for stretch and elasticity of titin and fibronectin.

The elastic protein titin comprises a tandem array of fibronectin type III and immunoglobulin domains, which are structurally similar 7-strand beta-sandwiches. A proposed mechanism for stretching titin, by sequential denaturation of individual fibronectin type III-immunoglobulin domains in response to applied tension, is analyzed here quantitatively. The folded domain is approximately 4 nm long, and the unraveled polypeptide can extend to 29 nm, providing a 7-fold stretch over the relaxed length. Elastic recoil is achieved by refolding of the denatured domains when the force is released. The critical force required to denature a domain is calculated to be 3.5-5 pN, based on a net free energy for denaturation of 7-14 kcal/mol, plus 5 kcal/mol to extend the polypeptide (1 cal = 4.184 J). This force is comparable to the 2- to 7-pN force generated by single myosin or kinesin molecules. The force needed to pull apart a noncovalent protein-protein interface is estimated here to be 10-30 pN, implying that titin will stretch internally before the molecule is pulled from its attachment at the Z band. Many extracellular matrix and cell adhesion molecules, such as fibronectin, contain tandem arrays of fibronectin type III domains. Both single molecules and matrix fibers should have elastic properties similar to titin.

Calorimetry↗

Out-of-plane propagation of elastic waves in two-dimensional phononic band-gap materials.

We have used a plane-wave-expansion model to study the out-of-plane propagation of elastic waves in a two-dimensional phononic band-gap material. The case of quartz rods embedded in an epoxy matrix has been computed. Band gaps for nonzero values of the wave-vector component parallel to the rods are shown to exist and are investigated. For wavelengths smaller than the period of the structure, modes are found that are localized in the epoxy intersites, and propagate perpendicularly to the plane of the structure.

Journal Article↗

Lichen planus limited to the nails in childhood: case report and literature review.

Although nail abnormalities have been reported to occur in 1% to 10% of patients with lichen planus, in children with lichen planus they are rarely mentioned in the literature. An 11-year-old boy had a two-month history of nail dystrophy affecting all the fingernails and the great toenails. The nail plates showed longitudinal ridging and thinning as well as onycholysis and distal splitting. There were no cutaneous or mucous membrane abnormalities. A nail biopsy specimen showed hyperkeratosis, hypergranulosis, and acanthosis in the ventral portion of the proximal nail fold and in the nail matrix. A band-like lymphocytic infiltrate was present in the superficial dermis, and the basal layer showed vacuolar alterations. A diagnosis of lichen planus was made. Treatment was intramuscular triamcinolone 20 mg once a month for six months. Since 1969 only 13 proved pediatric cases of lichen planus limited to the nails have been reported, including two children with 20-nail dystrophy and four with idiopathic atrophy of the nails.

Biopsy↗

Nanoliter serum sample analysis by mid-infrared spectroscopy for minimally invasive blood-glucose monitoring.

The aim of this study was to demonstrate that mid-infrared spectroscopy is able to quantify glucose in a serum matrix with sample volumes well below 1 muL. For this, we applied mid-infrared attenuated total reflectance (ATR) or transmission-based spectroscopic methods to glucose quantification in microsamples of dry-film sera, either undiluted or diluted 10 times in distilled water. The sample series spanned physiological glucose concentrations between 50 and 600 mg/dL and volumes of 80, 8, and 1 nL. Calibration was carried out using multivariate partial least-squares (PLS) modeling with spectral data between 1180 and 940 cm(-1). Best performance was achieved in the ATR experiments. For raw ATR spectra, the optimum standard error of prediction (SEP) of 13.3 mg/dL was obtained for the 8 nL sample series with subsequent 10-fold dilution. With respect to the coefficient of variation of the glucose assay, CV(pred), we obtained a value of 3% for the 80 nL volume samples with spectral preprocessing using matrix protein absorption bands as an internal standard, 4% for the 8 nL samples, and 6% for the 1 nL samples with raw data. Spectral standardization resulted in significant improvement, especially for the 80 nL volume sample series. By contrast, the accuracy of the glucose assay for the 1 nL sample volume series could not be improved either by internal standardization or by considering the dry film areas for normalization, which we attribute to varying topographies of the dry films.

Blood Chemical Analysis↗

[Intraepidermal mass of M. leprae in a case of seborrheic keratosis due to Hansen's disease (LLs)].

A 67-year-old patient has had exanthema in the lower right limb since 51 years ago (16 years old at onset), which underwent repeated remission and recurrence. At present, he has bilateral symmetrical widespread infiltrating exanthema and asymmetrical marked neuralhypertrophy, and has been diagnosed typical LLs (His father had the same disease). The exanthema recurred several years ago, and the patient is being treated for Hansen's disease. He had a dark brown flat elevation with a rough surface and the size of a small finger tip in his right abdominal skin for approximately 20 years. A biopsy was performed, and the specimen was fixed in 10% formalin and paraffin sections were prepared for histopathologic examination. A part of the specimen was processed forscanning electron microscopic examination. Seborrheic keratosis was diagnosed by H & E staining. Acid-fast (FITE) staining, immunohistochemical staining (keratin, S-100 protein, anti-PGL antibody and anti-BCG antibody) and scanning electron microscopy revealed the presence of bacteria (M. leprae) in the dermal foam cells, the matrix with a banded structure and the squamous epithelial cells which normally lack phagocytosis function. Compared to the basal cells of normal epidermis, the basal cells located adjacent to the dermis affected with seborrheic keratosis showed increased proliferation and more marked characteristics of a germinative cell. The degree of differentiation of the basal cells appeared regressed, and they probably possessed augmented phagocytic activity. The phagocytosed bacteria were probably carried by the epidermal cell cycle toward the surface layer. However, bacteria could not be found in the stratum corneum, probably due to an association with the lysosome.

Aged↗

Eggshell matrix proteins as defense mechanism of avian eggs.

This study focused on the role of eggshell matrix proteins as a function of potential natural antimicrobial defenses of avian eggs. The electrophoretic profile of SDS-PAGE showed that the soluble eggshell matrix proteins had three major bands of 15 000, 36 000, and 66 000 and several minor bands comprising 17 000, 25 000, 30 000, and 75 000, while insoluble matrix proteins were consisting of various bands comprising at least 16 distinct migration bands between 10 000 and 200 000. Three bacteria species, Pseudomonas aureginosa, Bacillus cereus, and Staphylococcus aureus, were found to be inhibited in the presence of soluble eggshell matrix proteins (100 microg/mL). On the other hand, Escherichia coli and Salmonella enteritidis were weakly inhibited at only an early stage of incubation time (up to 4 h). Scanning electron microscopy revealed that eggshell matrix proteins might interact and disrupt the membrane integrity of bacteria. The present study clearly indicated that avian eggshell matrix proteins possess a potential of novel antimicrobial defensin mechanism.

Animals↗

Changes in cells's secretory organelles and extracellular matrix during endochondral ossification in the mandibular condyle of the growing rat.

The mandibular condyle from 20-day-old rats was examined in the electron microscope with particular attention to intracellular secretory granules and extracellular matrix. Moreover, type II collagen was localized by an immunoperoxidase method. The condyle has been divided into five layers: (1) the most superficial, articular layer, (2) polymorphic cell layer, (3) flattened cell layer, (4) upper hypertrophic, and (5) lower hypertrophic cell layers. In the articular layer, the cells seldom divide, but in the polymorphic layer and upper part of the flattened cell layer, mitosis gives rise to new cells. In these layers, cells produce two types of secretory granules, usually in distinct stacks of the Golgi apparatus; type a, cylindrical granules, in which 300-nm-long threads are packed in bundles which appear "lucent" after formaldehyde fixation; and type b, spherical granules loaded with short, dotted filaments. The matrix is composed of thick banded "lucent" fibrils in a loose feltwork of short, dotted filaments. The cells arising from mitosis undergo endochondral differentiation, which begins in the lower part of the flattened cell layer and is completed in the upper hypertrophic cell layer; it is followed by gradual cell degeneration in the lower hypertrophic cell layer. The cells produce two main types of secretory granules: type b as above; and type c, ovoid granules containing 300-nm-long threads associated with short, dotted filaments. A possibly different secretory granule, type d, dense and cigar-shaped, is also produced. The matrix is composed of thin banded fibrils in a dense feltwork. In the matrix of the superficial layers, the "lucency" of the fibrils indicated that they were composed of collagen I, whereas the "lucency" of the cylindrical secretory granules suggested that they transported collagen I precursors to the matrix. Moreover, the use of ruthenium red indicated that the feltwork was composed of proteoglycan; the dotted filaments packed in spherical granules were similar to, and presumably the source of, the matrix feltwork. The superficial layers did not contain collagen II and were collectively referred to as perichondrium. In the deep layers, the ovoid secretory granules displayed collagen II antigenicity and were likely to transport precursors of this collagen to the matrix, where it appeared in the thin banded fibrils. That these granules also carried proteoglycan to the matrix was suggested by their content of short dotted filaments. Thus the deep layers contained collagen II and proteoglycan as in cartilage; they were collectively referred to as the hyaline cartilage region.

Actin Cytoskeleton↗

The effect of cooperative hydrogen bonding on the OH stretching-band shift for water clusters studied by matrix-isolation infrared spectroscopy and density functional theory.

Infrared spectra of the water clusters have been measured in the N2 + O2 matrix. The aggregation process of water in the matrix has been monitored by annealing the deposited samples up to 40 K and UV irradiation. The monomer, dimer, cyclic trimer and cyclic pentamer are found as water clusters in the matrix. For the hexamer, several structures such as chair, cage, prism, bag 1 and/or book 1 are likely to exist. By UV irradiation, the cyclic pentamer is predominantly formed from the monomer and dimer. On the other hand, by annealing the deposited sample, several hexamers are formed. The theoretical calculation for water clusters has revealed that the formation of one hydrogen bonding in a hydrogen-bonded chain cooperatively enhances or diminishes the strength of another hydrogen bond. Both proton donor (D) and acceptor (A) participating in a hydrogen-bonding pair DA are capable of forming hydrogen bonding with the other water molecules; D can additionally accept two protons and donate one proton, and A can additionally donate two protons and accept one proton. We have proposed the classification of hydrogen-bonding patterns considering the cooperativity, denoting as d'a'DAd''a'', where d and a are integers indicating the number of proton donors and acceptors to D (the single prime) and A (the double prime), respectively. Then, a magnitude given by MOH = -d' + a' + d'' - a'' has been introduced, which is very useful for connecting the hydrogen-bonding patterns to their OH wavenumbers. As a result, it is revealed that the OH stretching bands of water clusters are characterized by eight indicators (free and MOH = -2, -1, 0, 1, 2, 3 and 4). The classification proposed here is applicable to the OH band analysis for the hydrogen-bonded water and alcohols in a condensed phase.

Computer Simulation↗