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Comparative analysis of the fragments of highly repeated DNA of Homo, Pan, Macaca and Cercopithecus.

In the present paper the restriction patterns of Homo, Pan troglodytes, Macaca fascicularis and Cercopithecus aethiops, obtained by digestion with Eco RI endonuclease, were compared. All these species showed two common bands at 0.3 and 0.6 Kb. In this region another band at 0.5 Kb was present in Macaca fascicularis and Cercopithecus aethiops. Furthermore Macaca fascicularis, Cercopithecus aethiops and Pan troglodytes showed a series of bands not present in humans. The distance matrix shows a strong similarity between Homo and Pan and a decreasing similarity with Macaca fascicularis and Cercopithecus aethiops. No intraspecific variability was observed.

Animals↗

The migration of amphibian primordial germ cells in the chick embryo.

A fibrous band of extracellular materials on the chick embryo area pellucida/area opaca border is a preferential migratory pathway for chick embryo primordial germ cells (PGC). This band contains fibronectin, collagen Type I and sulphated glycosaminoglycans. It is known that PGCs from Xenopus laevis interact with fibronectin as they undergo migration in the embryo from their site of origin to the gonads. To establish whether this pathway is species specific in chick embryos it was decided to transplant PGC from Xenopus laevis embryos stage 48 on to chick embryos stage 4 fibrous band. Their rapid migration on this extracellular matrix and their subsequent re-orientation of the basement membrane has been studied by scanning electron microscopy.

Animals↗

Rovibrational and dynamical properties of the hydrogen bonded complex (CH2)2S-HF: a combined free jet, cell, and neon matrix-Fourier transform infrared study.

Fourier transform infrared spectra of the nu(s) (HF stretching) band of the (CH(2))(2)S-HF complex have been recorded at 0.1-0.5 cm(-1) resolution in a cooled cell, in a supersonic jet expansion seeded with argon and in a neon matrix at 4.5 K. The combination of controlled temperature effects over a range of 40-250 K and a sophisticated band contour simulation program allows the separation of homogeneous and inhomogeneous contributions and reveals significant anharmonic couplings between intramolecular and intermolecular vibrational modes similar to our previous work on (CH(2))(2)S-DF. The sign of the coupling constants is consistent with the expected strengthening of the hydrogen bond upon vibrational excitation of HF which also explains the observed small variations of the geometrical parameters in the excited state. The analysis of sum and difference combination bands involving nu(s) provides accurate values of intermolecular harmonic frequencies and anharmonicities and a good estimate of the dissociation energy of the complex. Frequencies and coupling parameters derived from gas phase spectra compare well with results from neon matrix experiments. The effective linewidth provides a lower bound for the predissociation lifetime of 10 ps. The comparison between effective linewidths and vibrational densities of states for (CH(2))(2)S-HF and -DF complexes highlights the important role of intramolecular vibrational redistribution in the vibrational dynamics of medium strength hydrogen bonds.

Journal Article↗

Pyrolysis of organomercury compounds: investigation by the method of matrix isolation.

The method of matrix isolation has been used to investigate mechanisms of gas-phase chemical reactions, in particular the pyrolysis of some organomercury compounds. A molecular beam of pyrolysis products was condensed simultaneously with a large excess of rare gas at temperatures from 5 to 15 degrees K to form a matrix that was subsequently studied by infrared spectroscopy. In the case of C(6)H(5)HgCCl(3), we found that pyrolysis in the temperature range 220-400 degrees C produced mainly dichlorocarbene. In addition, some trichloromethyl radical was observed and increased in relative importance at increased temperatures. Another identified product of pyrolysis was C(6)H(5)HgCl. In general, the same reactive intermediates, CCl(2) and CCl(3), were found from pyrolysis of Hg(CCl(3))(2) in the temperature range 250-500 degrees C, along with CCl(3)HgCl and HgCl(2). The identity of CCl(2) and CCl(2) was demonstrated by measurement of the relative intensities and isotopic splittings of stretching vibrations due to the chlorine isotopes. Isotopic patterns found for CCl(2) are: v(3) (745.8, 744.0, 741.8 cm(-1)), v(1) (719.5, 717.0, 714.9 cm(-1)) and for CCl(3) are: v(3) (897.8, 896.4, 895.2, 893.9 cm(-1)). Less dilution with the rare gas or warming of the matrix produced a decrease of CCl(2) and CCl(3) spectral bands and an increase of bands due to C(2)Cl(4), C(2)Cl(6), and other products. These results show the usefulness of matrix isolation in the study of such reactive species as CCl(2) produced by pyrolysis in the gas phase.

Journal Article↗

The terminal tendon of the digital extensor mechanism: Part I, anatomic study.

PURPOSE: The purpose of this study was to examine the anatomy of the terminal tendon (TT) and its relationship to adjacent structures. MATERIALS: The extensor tendons of 56 cadaveric digits (52 fresh-frozen and 4 formalin-preserved) underwent anatomic dissection. RESULTS: The TT is a segment between the convergence of the lateral bands proximally and the bony insertion in the phalanx distally. The ulnar lateral bands were thicker than the radial bands. The average distance from the TT insertion to the germinal matrix of the nail bed was 1.4 mm. The triangular ligament (TL) is a thin layer of transverse fibers between the lateral bands proximal to the TT. The dimensions of the TT and the TL vary relative to the size of the digit with the largest often occurring in the middle, followed by the ring, index, and small finger. The transverse retinacular ligament (TRL) had a dorsal attachment to the lateral bands and was much more defined and distinct than the oblique retinacular ligament. The ulnar TRL often was thicker than the radial TRL. CONCLUSIONS: The TT is the primary structure responsible for extending the distal interphalangeal (DIP) joint. The function of the adjacent retinacular structures is to provide stability to the TT. The thin nature and proximity of the TT to the nail matrix must be kept in mind during surgery. Knowledge of the TT anatomy is necessary for further study of its kinematics and pathology and for diagnosis and management of its disorders.

Aged↗

Electronic spectroscopy of UO2 isolated in a solid Ar matrix.

Dispersed fluorescence spectra have been recorded for UO(2) isolated in a solid Ar matrix. Near UV excitation produced groups of emission bands in the 370-420 and 465-645 nm spectral regions. These bands originated from two energetically close upper levels and terminated on a range of low-lying electronic states. Comparisons with electronic structure calculations indicate that the ground and low-lying electronic states of UO(2) are derived from the U(5f7s) configuration. The level of agreement between the observed and predicted electronic energies provides an impressive validation of the calculations. In previous studies of matrix isolated UO(2), it had been suggested that the ground state in solid Ar is U(5f(2)) (3)H. The present results do not appear to be consistent with this hypothesis.

Journal Article↗

Acquired drug resistance is accompanied by modification in the karyotype and nuclear matrix of a rat carcinoma cell line.

A Walker 256 breast carcinoma cell line (WR) exhibiting a greater than 20-fold resistance to alkylating agents has been selected from a parent cell line (WS). Karyotypic heterogeneity was apparent, with a number of differences evident between WR and WS cells. The modal chromosome number for WS is 62; for WR, 54; double minutes were found only in WR, whereas spontaneous chromosomal aberrations were present in approx. 40% of the WS cells. No similar aberrations were observed in WR. Using SDS-gel electrophoresis and subsequent silver staining, differences in the profile of nuclear matrix proteins in WR and WS were observed. A diffuse band at approx. 70 kD in the WS was absent in WR cells. This protein was phosphorylated, together with a number of the other major matrix polypeptides. Levels of phosphorylated matrix proteins were approximately equivalent in both WR and WS cell lines, but matrix protein phosphorylation levels were approx. 2-fold higher than corresponding values for bulk nuclear proteins. Selective pressure of drug exposure has resulted in enhanced genetic stability in WR cells and observed karyotype differences are accompanied by modifications in the structural proteins of the nuclear matrix. Whether the observed differences are the cause or result of drug resistance remains to be established.

Animals↗

Stop-pass behavior of acoustic waves in a 1D fractured system

This study examines the dispersion and the stop-pass band behavior of acoustic waves propagating across periodically spaced and non periodically spaced parallel fractures. Laboratory ultrasonic wave measurements performed on a stack of synthetic fractures (identical steel plates with roughened interfaces) and numerical propagator matrix simulations show spectra with distinct stop-pass band structures that develop with decreasing fracture stiffness. To understand the physics behind these observations, an exact dispersion equation for wave propagation through an infinite series of equally spaced fractures is derived using displacement-discontinuity boundary conditions to model the constitutive behavior of the fractures and Floquet's (Bloch's) theory for the periodic boundary conditions. Both the measured and numerically simulated stop-pass band structures show good agreement with the theoretical predictions. Furthermore, the theory reveals that the left boundary of the stop-bands contains information about the fracture stiffness, suggesting the possibility of determining the stiffness of the parallel fractures from seismic waves. This paper also discusses the effects of fractured systems with random distributions of fracture spacings and stiffnesses on the stop-pass band structures of seismic waves in fractured rock.

Journal Article↗

Difference in gene expression for matrix metalloproteinase-1 between early and advanced hepatocellular carcinomas.

The histological observation that well-differentiated cancer cells in early hepatocellular carcinoma (HCC) invade portal tracts and/or fibrous bands and that these fibrous tissues then disappear suggests the participation of matrix metalloproteinase-1 (MMP-1) in the degradation of fibrous tissue. To confirm this hypothesis, the authors investigated the localization of both the MMP-1 protein and its messenger RNA (mRNA) in early HCC immunohistochemically and by in situ hybridization using complementary DNA (cDNA) and synthetic antisense probe of MMP-1; they then compared the results with those in advanced HCC. MMP-1 gene transcripts and protein were observed in well-differentiated cancer cells of early HCC but not in moderately or poorly differentiated cancer cells. Thus, cancer cells producing MMP-1 in early HCC may destroy the portal tract tissue adjacent to the cancer lesion and/or the fibrous band of cirrhosis. These results seem to have demonstrated a difference in the mechanism of cancer growth and invasion between early and advanced HCCs.

Adult↗

Quantitation of adenovirus type 5 empty capsids.

Adenovirus empty capsids are immature intermediates that lack DNA and viral core proteins. Highly purified preparations of empty and full capsids were generated by subjecting purified adenovirus preparations to repeated cesium chloride gradient separations. PAGE results revealed that empty capsids contain at least five bands that correspond to proteins absent from the mature virus proteome. Peptide mapping by matrix-assisted laser desorption/ionization time-of-flight MS revealed that three of these bands correspond to varying forms of L1 52/55kDa, a protein involved in the encapsidation of the viral DNA. One band at around 31kDa was found to include precursors to proteins VI and VIII. These precursors correspond to proteins that have not been cleaved by the adenovirus-encoded protease and are not present in the mature full capsids. The precursor to protein VIII (pVIII), a capsid cement protein, is used in this study as a marker in reverse-phased HPLC (RP-HPLC) analyses of adenovirus for the quantitation of empty capsids. A novel calculation method applied to the integration of RP-HPLC chromatograms allowed for the generation of a percentage empty capsid value in a given adenovirus preparation. The percentage empty capsid values generated to date by this method show a high degree of precision and good agreement with a cesium chloride gradient/SDS-PAGE quantitation method of empty capsids. The advantage of this method lies in the accurate, precise, and rapid generation of the percentage of empty capsids in a given purified virus preparation without relying on tedious and time-consuming cesium chloride gradient separations and extractions.

Adenoviridae↗

Isolated lichen planus of the nails treated with etretinate.

A 46-year-old Japanese man with isolated lichen planus of the nails is reported. The affected nails were all of the fingernails and the third and fourth toenails of both feet. Histologically, the ventral part of the proximal nail folds showed mild band-like cell infiltration below the epidermis, and the nail beds (ventral matrix) showed hypergranulosis and epidermal-dermal interphase activity with liquefaction degeneration of basal cells and a band-like cell infiltration consisting of lymphoid cells and histiocytes. Systemic use of etretinate in combination with application of steroid lotion was evaluated as very effective. Although the use of retinoids for lichen planus of the nails remains only a suggested method of treatment, a positive trial should be attempted, since this disorder sometimes results in severe nail damage.

Etretinate↗

Reduction of ischemic myocardial damage in the dog by lidocaine infusion.

The effects of lidocaine infusion on the ultrastructural damage induced in cardiac muscle by normothermic cardiopulmonary bypass were assessed in 15 dogs. Six dogs received no medication other than sodium pentobarbital (25 mg/kg, intravenously) while 9 dogs were treated with lidocaine after anesthesia. Lidocaine was given as a 2-mg/kg loading dose 10 minutes prior to ischemic arrest and a 2-mg/min continuous infusion during the entire experimental period. Biopsy samples of the left ventricular apex were taken 15 and 45 minutes after the start of ischemic arrest and 5 minutes after resumption of coronary blood flow. Biopsy samples were also obtained from 4 animals after thoracotomy to serve as controls for experimental procedures. Myocardial ultrastructure in the 4 control animals was comparable to that reported by other investigators. Five of 6 of the nontreated dogs and 8 of 9 lidocaine-treated dogs survived the entire period of ischemia and 5 minutes of coronary reperfusion. However, the extent of ultrastructural damage varied considerably between the two groups. In the experimental dogs receiving no lidocaine, mitochondria were swollen, cristae were absent, the mitochondrial matrix was cleared, and sarcomeres were disrupted. Myelin figures and contraction bands were also observed. None of the surviving lidocaine-treated animals had ultrastructural changes comparable to the worst ones in nontreated dogs. Damage was limited to some swelling of mitochondria with focal clearing of matrix. Most cristae remained intact. There were no myelin figures and few contraction bands. The results suggest that lidocaine protects the integrity of ischemic myocardium. It is suggested that this protection resulted from stabilization of plasma and/or mitochondrial membranes. (Am J Pathol 87:399-414, 1977).

Animals↗

Hensin, a new collecting duct protein involved in the in vitro plasticity of intercalated cell polarity.

Two forms of intercalated cells are present in kidney collecting tubules, the alpha cell has apical endocytosis, apical H+-ATPase and basolateral band 3, while beta cells have reversed polarity of these proteins and no apical endocytosis. When a beta cell line was seeded at high density, it changed into the alpha form. We previously showed that a partially purified 230 kD extracellular matrix protein of high density cells was able to retarget band 3 from apical to basolateral domains and stimulated apical endocytosis in vitro (Van Adelsberg, J., J.C. Edwards, J. Takito, B. Kiss, and Q. Al-Awqati. 1994. Cell. 76:1053-1061). We now purify this protein, which was named hensin, to near homogeneity and find that it belongs to the macrophage scavenger receptor cysteine rich (SRCR) family. An antibody, generated against a fusion protein made from a partial cDNA recognized a 230-kD protein in rabbit kidney and in the intercalated cell line. In vitro, the hensin antibody inhibited expression of apical endocytosis. Hensin was secreted in a polarized manner and bound to the basolateral membrane and extracellular matrix. Immunohistochemistry of the kidney showed that it was expressed only in collecting tubules. Double immunofluorescence with hensin and peanut lectin, H+-ATPase, or band 3 showed many patterns; most alpha-cells had hensin staining while 50% of beta-cells did not. These results suggest that hensin may also be involved in the polarity reversal of intercalated cells in vivo.

Amino Acid Sequence↗

A new infrared electronic transition of the oxidized primary electron donor in bacterial reaction centers: a way to assess resonance interactions between the bacteriochlorophylls.

The primary electron donor in the reaction center of purple photosynthetic bacteria consists of a pair of bacteriochlorophylls (PL and PM). The oxidized dimer (P+) is expected to have an absorption band in the mid-IR, whose energy and dipole strength depend in part on the resonance interactions between the two bacteriochlorophylls. A broad absorption band with the predicted properties was found in a previously unexplored region of the spectrum, centered near 2600 cm-1 in reaction centers of Rhodobacter sphaeroides and several other species of bacteria that contain bacteriochlorophyll a, and near 2750 cm-1 in Rhodopseudomonas viridis. The band is not seen in the absorption spectrum of the monomeric bacteriochlorophyll cation in solution, and it is missing or much diminished in the reaction centers of bacterial mutants that have a bacteriopheophytin in place of either PL or PM. With the aid of a relatively simple quantum mechanical model, the measured transition energy and dipole strength of the band can be used to solve for the resonance interaction matrix element that causes an electron to move back and forth between PL and PM, and also for the energy difference between states in which a positive charge is localized on either PL or PM. (The absorption band can be viewed as representing a transition between supermolecular eigenstates that are obtained by mixing these basis states.) The values of the matrix element obtained in this way agree reasonably well with values calculated by using semiempirical atomic resonance integrals and the reaction center crystal structures.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriochlorophylls↗

Changes in the protein components of rat incisor enamel during tooth development.

Enamel-matrix components from rat incisor enamel were extracted from tissue at different stages of development on single teeth. Separations of proteins using urea and SDS acrylamide gel electrophoresis were compared. The bulk of the matrix exhibited SDS mol. wt of 25-30,000 with smaller amounts at approximately 18,000 and about 10-12,000. Trace amounts of material at -50,000 and 70,000 were detected. These were presumably associated with the mineral phase as their yield increased after demineralization. The proportion of small molecular weight components increased with tissue age. Using urea, many more proteins were separated (up to 20) into fast, intermediate and slowly-migrating components. Disappearance of small bands of intermediate mobility at the end of matrix secretion suggested that they were early ameloblast products which were rapidly degraded after secretion. Both slowly- and rapidly-migrating components increased with tissue age indicating progressive degradation of parent molecules of intermediate mobility into highly charged and relatively uncharged molecules.

Animals↗

Analysis of bone morphogenetic protein (BMP) derived from human and bovine bone matrix.

Recently, Bone Morphogenetic Protein (BMP) has attracted the attention of a number of investigators, but its elucidation remains incomplete. At present, the determination of its amino acid sequence, which is necessary for its synthesis, and screening for a carrier that allows BMP to be effective in small amounts are unsolved problems. Bone morphogenetic protein is studied here to clarify its clinical applications. BMP was extracted from human and bovine bone matrix with 4 M guanidine-HCl and purified by liquid chromatography. Acrylamide electrophoresis (SDS-PAGE) and isoelectric focusing (IEF) showed that the purified BMP was homogeneous. We used type I collagen as the carrier in the bioassay. This BMP induced new bone in situ three weeks after implantation in muscle pouches in Wistar rats. The molecular weights of human and bovine bone matrix-derived BMP are 17.0 and 18.0 kDa by SDS-PAGE, and pI values for both are 4.9 by IEF. Human and bovine bone matrix-derived BMP are peptides containing 165 and 163 amino acids, respectively, according to amino acid analysis. The NH2-terminal sequence of bovine bone matrix-derived BMP was obtained from the bovine band, electroblotted onto polyvinylidene difluoride membrane, that corresponded to the final purified fraction. The sequence differs from previously designated BMPs25 and other proteins reported to have similar activity, but the physicochemical characteristics are comparable to the native preparations.

Amino Acid Sequence↗

Recurrent coamplification of cytoskeleton-associated genes EMS1 and SHANK2 with CCND1 in oral squamous cell carcinoma.

Chromosomal band 11q13 is frequently amplified in oral squamous cell carcinoma (OSCC) and assumed to be critically involved in tumor initiation and progression by proto-oncogene activation. Though cyclin D1 (CCND1) is supposed to be the most relevant oncogene, several additional putative candidate genes are inside this chromosomal region, for which their actual role in tumorigenesis still needs to be elucidated. To characterize the 11q13 amplicon in detail, 40 OSCCs were analyzed by comparative genomic hybridization to DNA microarrays (matrix-CGH) containing BAC clones derived from chromosomal band 11q13. This high-resolution approach revealed a consistent amplicon about 1.7 Mb in size including the CCND1 oncogene. Seven BAC clones covering FGF3, EMS1, and SHANK2 were shown to be frequently coamplified inside the CCND1 amplicon. Subsequent analysis of tissue microarrays by FISH revealed amplification frequencies of 36.8% (88/239) for CCND1, 34.3% (60/175) for FGF3, 37.4% (68/182) for EMS1, and 36.3% (61/168) for SHANK2. Finally, quantitative mRNA expression analysis demonstrated consistent overexpression of CCND1 in all tumors and of EMS1 and SHANK2 in a subset of specimens with 11q13 amplification, but no expression of FGF3 in any of the cases. Our study underlines the critical role of CCND1 in OSCC development and additionally points to the functionally related genes EMS1 and SHANK2, both encoding for cytoskeleton-associated proteins, which are frequently coamplified with CCND1 and therefore could cooperatively contribute to OSCC pathogenesis.

Carcinoma, Squamous Cell↗

Observing growth steps of collagen self-assembly by time-lapse high-resolution atomic force microscopy.

Insights into molecular mechanisms of collagen assembly are important for understanding countless biological processes and at the same time a prerequisite for many biotechnological and medical applications. In this work, the self-assembly of collagen type I molecules into fibrils could be directly observed using time-lapse atomic force microscopy (AFM). The smallest isolated fibrillar structures initiating fibril growth showed a thickness of approximately 1.5 nm corresponding to that of a single collagen molecule. Fibrils assembled in vitro established an axial D-periodicity of approximately 67 nm such as typically observed for in vivo assembled collagen fibrils from tendon. At given collagen concentrations of the buffer solution the fibrils showed constant lateral and longitudinal growth rates. Single fibrils continuously grew and fused with each other until the supporting surface was completely covered by a nanoscopically well-defined collagen matrix. Their thickness of approximately 3 nm suggests that the fibrils were build from laterally assembled collagen microfibrils. Laterally the fibrils grew in steps of approximately 4 nm, indicating microfibril formation and incorporation. Thus, we suggest collagen fibrils assembling in a two-step process. In a first step, collagen molecules assemble with each other. In the second step, these molecules then rearrange into microfibrils which form the building blocks of collagen fibrils. High-resolution AFM topographs revealed substructural details of the D-band architecture of the fibrils forming the collagen matrix. These substructures correlated well with those revealed from positively stained collagen fibers imaged by transmission electron microscopy.

Animals↗