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Radiation-induced graft copolymer SBS-g-VP for biomaterial usage.

The grafting of 4-vinyl pyridine (VP) to styrene-butadienestyrene triblock copolymer (SBS) by radiation-induced graft copolymerization was studied. The cohesive properties such as cohesive energy (Ecoh), molar volume (V), solubility parameter (delta), molar molecular weight (WM), specific volume (Vg), and density (1/Vg) of SBS-g-VP graft copolymer were calculated according to the group contribution of Fedors. The morphology of SBS-g-VP was studied by optical polarizing microscopy. We also measured the glass transition temperature and the mechanical properties of SBS-g-VP graft copolymer. Contact angle and blood-clotting time experiments were also performed to evaluate the biocompatibility of SBS-g-VP. A second domain was found in the SBS-g-VP graft copolymer, which resulted in different properties between SBS-g-VP and SBS. The blood compatibility of SBS-g-VP as measured by the Lee-White clotting test was better than that of SBS and polystyrene.

Biocompatible Materials↗

Physical characteristics and chemical degradation of amorphous quinapril hydrochloride.

This study was designed to investigate the relationships between the solid-state chemical instability and physical characteristics of a model drug, quinapril hydrochloride (QHCl), in the amorphous state. Amorphous QHCl samples were prepared by rapid evaporation from dichloromethane solution and by grinding and subsequent heating of the crystalline form. Physical characteristics, including the glass transition temperature and molecular mobility, were determined using differential scanning calorimetry, thermogravimetric analysis, powder x-ray diffractometry, polarizing microscopy, scanning electron microscopy, and infrared spectroscopy. The amorphous form of QHCl, produced by both methods, has a T(g) of 91 degrees C. Isothermal degradation studies showed that cyclization of QHCl occurred at the same rate for amorphous samples prepared by the two methods. The activation energy was determined to be 30 to 35 kcal/mol. The rate of the reaction was shown to be affected by sample weight, dilution through mixing with another solid, and by altering the pressure above the sample. The temperature dependence for chemical reactivity below T(g) correlated very closely with the temperature dependence of molecular mobility. Above T(g), however, the reaction was considerably slower than predicted from molecular mobility. From an analysis of all data, it appears that agglomeration and sintering of particles caused by softening of the solid, particularly above T(g), and a resulting reduction of the particle surface/volume ratio play a major role in affecting the reaction rate by decreasing the rate of removal of the gaseous HCl product.

Angiotensin-Converting Enzyme Inhibitors↗

Conjugated organometallic polymers containing Vollhardt's cyclobutadiene complex: aggregation and morphologies.

Organometallic polymers were prepared by acyclic diyne metathesis (ADIMET) or by Pd-catalyzed coupling of 1,3-diethynylcyclobutadiene(cyclopentadienyl)cobalt with a suitably substituted diiodobenzene. The polymers obtained by Heck coupling show a degree of polymerization (Pn) of 20-60. The monomers for ADIMET were made by the Pd-catalyzed coupling of [1,3-bis(trimethylsilylethynyl)-2,4-bis(trimethylsilyl)cyclobutadiene](cyclopentadienyl)cobalt to 1-bromo-2,5-dialkyl-4-propynylbenzenes in the presence of KOH in yields of 40-48%. The monomers carry hexyl, ethylhexyl, and (S)-3,7-dimethyloctyl side chains. Polymerization of the propynylated monomers furnishes organometallic polymers with a Pn of up to 230 arylene-ethynylene units. The polymers were fully characterized by polarizing microscopy, transmission electron microscopy, circular dichroism, differential scanning calorimetry, and X-ray diffraction (XRD). They show nematic, lyotropic liquid crystalline phases as well as chiroptical properties from which aggregation in poor solvents and in the solid state can be concluded. Lamellar or irregular honeycomb-shaped morphologies in these organometallic polymers can be detected by electron microscopy.

Journal Article↗

Inherited amyloid polyneuropathy type IV (gelsolin variant) in a Japanese family.

We describe a Japanese family with familial amyloid polyneuropathy type IV. The family originates from central Japan, Nagano prefecture, and is unrelated to Finnish or other Caucasian populations. Of 42 members in three generations, 14 individuals (5 men, 9 women) are affected by corneal lattice dystrophy, cranial neuropathy, mild peripheral neuropathy, and skin changes. Polarizing microscopy and immunohistochemistry studies of skin biopsy samples demonstrated abundant amyloid deposits, which bound an antigelsolin monoclonal antibody. Direct sequence analysis of a DNA fragment spanning codon 187 of plasma gelsolin complementary DNA and restriction analysis using a modified polymerase chain reaction demonstrated a single base substitution, guanine to adenine, at nucleotide position 654, which is identical to the mutation in Finnish familial amyloid polyneuropathy type IV. This strongly suggests that the mutation causes the familial amyloid polyneuropathy type IV phenotype regardless of ethnic background.

Adult↗

Hominid cranial bone structure: a histological study of Omo 1 specimens from Ethiopia using different microscopic techniques.

The microstructure of a hominid cranial vault has not previously been studied to determine its tissue histology, and differences in comparison with that of modern humans. We selected the parietals of Omo-Kibish 1, regarded as one of the oldest (about 130,000 years old) anatomically modern humans, and Omo 1 (Howell), which is a very recent human (about 2,000 years old)-both from the same area of Ethiopia. A combination of macrophotography, polarizing microscopy in the incident and transmission illumination mode, and confocal laser scanning microscopy (CLSM) was employed to examine thin sections, as well as polished and unpolished block faces of unembedded bone fragments, to minimize specimen destruction as much as possible. The methods enabled remarkably detailed information on bone microstructure and remodeling to be gleaned from tiny fragments of bone. The best method for examining fossilized human bones was shown to be that of incident light microscopy, which was the least destructive while producing the most amount of information. Unless the above methods are used, bone-filling minerals, such as calcite, can cause erroneous estimations of bone thickness, as observations with the naked eye or even a magnifying glass cannot determine the limit between the cortex and the diploe. This is particularly important for sciences such as paleoanthropology, in which, for instance, a thick cranial bone of Homo erectus may be confused with a pathological one of H. sapiens and vice versa. Cross sections of parietal bones revealed differences between Omo-Kibish 1 and Omo 1 (Howell) in diploic histology and in the relative thickness between the cortex and diploe, with the former specimen having an H. erectus ratio despite its H. sapiens gross anatomy. Omo-Kibish 1 may still retain some affinities with H. erectus despite its being classified as H. sapiens. Newly described histological structures, such as the reverse type II osteons, the multicanalled osteons, and the osteocytomata are presented here. A modern human skeletal anatomy does not necessarily imply a modern human cranial bone histology. The outer circumferential lamellae of cranial bones are in essence growth lines. Cranial histology of hominids may provide useful information concerning their taxonomy and life history, including such factors as growth rate, developmental stress, and diet.

Animals↗

Polarized light microscopic analyses of collagen fibers in the rat incisor periodontal ligament in relation to areas, regions, and ages.

We prepared decalcified sagittal sections (20 microm thick) from the incisal, middle, and basal regions of the mandibular incisor of male Wistar rats aged 2, 6, 12, and 24 months, and examined the sections using polarized light microscopy. Most of the birefringent fibers appeared to run obliquely across the periodontal ligament. Birefringent fibers running parallel to the long axis of the incisor were also found in the intermediate area of the ligament. Similar fiber architecture was observed in all four age groups. Quantitative analysis showed that the retardation values of collagen were higher in the bone- and tooth-related areas and lower in the intermediate area of the ligament. The values for the bone- and tooth-related areas increased from the basal toward the incisal regions in all four age groups. Age-related changes in the retardation values were found only in the incisal region of the incisor. In the incisal region, the values for the bone- and tooth-related areas increased markedly from 2-24 months of age, whereas those for the intermediate area increased slightly but significantly with age. Our findings indicate that the degrees of molecular organization and alignment of collagen fibers in the bone- and tooth-related areas of the ligament are higher than those in the intermediate area and increase near the incisal region and with age. It is also suggested that the collagen fibers in the intermediate area remain immature along the long axis of the incisor throughout the life span of the animal.

Age Factors↗

The effects of pelvic neurectomy on collagen in the cervix of the pseudopregnant rat.

Bilateral section of the pelvic parasympathetic nerves (pelvic-neurectomy) on day 5 of pseudopregnancy had no effect on the wet weight of ovaries, uteri, and/or cervices, but at day 9 serum progesterone was reduced to approximately half that of sham-operated animals. Collagen in the cervix was visualized with picrosirius red staining under polarization microscopy. Pelvic neurectomy decreased the birefringence of Type 1 collagen in the cervix to less than half that of sham operated animals. The decreased birefringence, an index of the organization of collagen fibers, is believed to be attributable to reduced progesterone levels. Alternatively, the pelvic nerve may directly influence mucopolysaccharides or collagenolytic enzymes in the cervix.

Animals↗

Intramembranous bone matrix is osteoinductive.

All known bone-derived osteoinductive factors have been isolated from endochondral (EC) bones and all initiate bone induction via EC ossification. However, to date no attempt has been made to isolate comparable factors from bones which form initially and completely via intramembranous (IM) ossification. The purpose of this work was to isolate osteoinductive proteins from IM bones. To accomplish this, we extracted proteins from bovine frontal bone matrix (intramembranous origin) using methods previously described for endochondral (EC) bone matrix (i.e., femur). Bone powder (< 1 mm) was decalcified and proteins extracted with 4 M guanidine hydrochloride. Ultrafiltration was used to isolate and concentrate a 10-100 kilodalton (kDa) fraction, upon which heparin-Sepharose (HS) affinity chromatography was performed. HS-binding (HS-B) and non-binding proteins (HS-NB) were lyophilized with bovine type I collagen (Vitrogen) to form pellets which were implanted subcutaneously in rats. Radiology as well as brightfield, fluorescent, and polarizing microscopy were used to assess the formation of ectopic bone at the site of pellet implantation. In this report we demonstrate that a heparin-Sepharose binding, osteoinductive factor can be extracted and partially purified from bovine intramembranous bone matrix. This factor has a different sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) banding pattern than a comparable osteoinductive/chondroinductive factor isolated from EC bone.

Animals↗

Acute gout presenting in the manubriosternal joint.

A 54-year-old black man developed acute pain and swelling of the manubriosternal joint. Acute gouty arthritis was diagnosed by arthrocentesis and polarizing microscopy. The histology of the manubriosternal joint and its involvement in other arthritides are briefly discussed.

Arthritis↗

Immediate "steroid flare" from intraarticular triamcinolone hexacetonide injection: case report and review of the literature.

We describe a patient who had an immediate, intense, localized synovitis due to intraarticular triamcinolone hexacetonide injection. The reaction was secondary to rapid intracellular ingestion of the steroid microcrystals as demonstrated by compensated polarized microscopy. We report the unique nature of this patient's response, and we review previous literature regarding "steroid flare" after intraarticular injection.

Adult↗

Amyloidosis in Behçet's syndrome.

We investigated the frequency of amyloidosis in 99 consecutive patients (65 men and 34 women) with Behçet's syndrome. Rectal biopsies performed on all patients showed no amyloidosis by polarizing microscopy. The results of urinalyses were normal in all patients. Among 1,130 patients registered in our Behçet's syndrome outpatient clinic, 5 were found to have systemic amyloidosis, all of the AA type.

Adolescent↗

Helical supramolecular aggregates based on ureidopyrimidinone quadruple hydrogen bonding.

A series of mono- and bifunctional compounds 2-7, based on the ureido pyrimidinone quadruple hydrogen bonding unit, was prepared to study the mode of aggregation of these compounds in the bulk and in solution. Compounds 2-7 exhibit thermotropic liquid crystalline properties, as evidenced by differential scanning calorimetry and optical polarization microscopy. The presence of an ordered hexagonal discotic (D(ho)) phase of 2 a was confirmed by X-ray diffraction on an aligned sample. In chloroform, the bifunctional compounds form cyclic dimers at millimolar concentrations, and these dimers exist in equilibrium with linear species above a critical concentration, which may be from 6 mM to greater than 260 mM, depending on the structure of the spacer. Circular dichroism measurements in chloroform did not show a Cotton effect. Dodecane solutions of compounds 3, 4 b, and 7 b display a Cotton effect at the absorption band of the phenyl-pyrimidinone unit. Amplification of chirality was observed in mixtures of 7 a and 7 b, but not in mixtures of 4 a and 4 b, indicating that 7 a and 7 b form mixed polymeric aggregates with a helical architecture in dodecane solution, whereas 4 a and 4 b do not. The Cotton effect is lost upon increasing the temperature. Half of the helicity is lost at 25 degrees C for 3 and at 60 degrees C for 4 b, suggesting that 3, bearing the shorter spacer, forms less stable columns than 4 b. Compound 7 b loses half of its helicity at 45 degrees C. Compounds 2 b, 5, and 6 do not exhibit helical organization, as evidenced by the absence of Cotton effects.

Calorimetry, Differential Scanning↗

The stabilization of microtubules in isolated spindles by tubulin-colchicine complex.

We have analyzed the effect of colchicine and tubulin dimer-colchicine complex (T-C) on microtubule assembly in mitotic spindles. Cold- and calcium-labile mitotic spindles were isolated from embryos of the sea urchin Lytechinus variegatus employing EGTA/glycerol stabilization buffers. Polarization microscopy and measurements of spindle birefringent retardation (BR) were used to record the kinetics of microtubule assembly-disassembly in single spindles. When isolated spindles were perfused out of glycerol stabilizing buffer into a standard in vitro microtubule reassembly buffer (0.1 M Pipes, pH 6.8, 1 mM EGTA, 0.5 mM MgCl2, and 0.5 mM GTP) lacking glycerol, spindle BR decreased with a half-time of 120 s. Colchicine at 1 mM in this buffer had no effect on the rate of spindle microtubule disassembly. Inclusion of 20 microM tubulin or microtubule protein, purified from porcine brain, in this buffer resulted in an augmentation of spindle BR. Interestingly, in the presence of 20 microM T-C, spindle BR did not increase, but was reversibly stabilized; subsequent perfusion with reassembly buffer without T-C resulted in depolymerization. This behavior is striking in contrast to the rapid depolymerization of spindle microtubules induced by colchicine and T-C in vivo. These results support the current view that colchicine does not directly promote microtubule depolymerization. Rather, it is T-C complex that alters microtubule assembly, by reversibly binding to microtubules and inhibiting elongation. In vivo, colchicine can induce depolymerization of nonkinetochore spindle microtubules within 20 s. In vitro, colchicine blocks further microtubule assembly, but does not induce rapid disassembly.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Chromatin supraorganization and extensibility in mouse hepatocytes following starvation and refeeding.

BACKGROUND: The effect of 48 h of starvation and of 48 h of refeeding subsequent to starvation on chromatin supraorganization and extensibility was studied in hepatocytes from adult mice. METHODS: Methods used involved topochemical assays, image analysis, gravity action, and polarization microscopy. RESULTS: Starvation increased the chromatin packing states, especially in areas of noncondensed chromatin, and induced drastic decreases in concanavalin A reactivity due to nuclear matrix glycoproteins and the frequency of nuclei with chromatin extensibility under gravity. Changes in chromatin packing state were accompanied by shifts of nuclear areas of part of the nuclear population to smaller values but did not affect the respective Feulgen-DNA amounts except for a few nuclei. The extent of chromatin unpackaging, but not of frequency of nuclei with formation of extended chromatin fibers, in starved mice that were refed was greater than in well-fed controls. Refeeding induced increase in Feulgen-DNA amounts and regain and redistribution of concanavalin A-reactive nuclear glycoproteins. However, the duration of refeeding used was probably insufficient to reestablish the stereo arrangement of the chromatin-nuclear matrix and to restore chromatin fluidity to the level seen in well-fed mice. CONCLUSIONS: The changes in the liver cell nuclei associated with starvation and refeeding of adult mice involved chromatin supraorganization, hepatocyte proliferation (refeeding), and the loss, regain, and redistribution of nuclear proteins, especially nuclear matrix components, related to chromatin organization and extensibility. These changes are suggested as favoring the silencing and reactivation of transcriptional activities, depending on the organism's nutritional state.

Animals↗

Chromatin supraorganization and extensibility in mouse hepatocytes with development and aging.

BACKGROUND: Chromatin supraorganization and extensibility, which lead to the formation of extended chromatin fibers (ECF), are affected by starvation and refeeding in adult mouse hepatocytes. It is expected that they could also change with mouse development and aging. METHODS: Methods used involved topochemistry, image analysis, microspectrophotometry, gravity action, and polarization microscopy. RESULTS: Increased nuclear areas and Feulgen-DNA amounts with advancing hepatocyte polyploidy were found with development and aging. A slightly less packed chromatin with more heterogeneously distributed condensation levels was detected in young and old mice. Con-A responsiveness was almost absent in young mice but very deep in aged mice. ECFs formed from nuclei of adult and aged mice but not from nuclei of young mice. The frequency of ECF formation with the long lysis protocol increased with aging. CONCLUSIONS: In young mice, a less packed chromatin state may be associated with more intense gene activity, thus increasing the DNA-nuclear matrix interactions, and inhibiting ECF formation. Reduced DNA-nuclear matrix interactions besides defects in heterochromatin formation may induce higher ECF formation and chromatin unpackaging in old mice. We suggest that differences in Con-A staining relate to different gene activity with advancing development and aging.

Aging↗

Unsuspected systemic amyloidosis diagnosed by fine-needle aspiration of the salivary gland: case report.

Amyloidosis of the head and neck region may represent a local amyloidoma or a manifestation of systemic disease. Involvement of major salivary glands by either primary or secondary forms of amyloidosis is very rare. We describe a case of systemic amyloidosis that initially presented as submandibular gland mass and was diagnosed by fine-needle aspiration (FNA). A 69-year-old male presented with submandibular mass. His past medical history was significant for left forearm melanoma that was excised 6 years ago and tricuspid valve endocarditis after valvular replacement 3 months prior to FNA of the submandibular gland. The patient had no symptoms or clinical and laboratory data suggestive of amyloidosis. FNA specimen showed salivary gland tissue and abundant amorphous material, which stained positive for amyloid with Congo red stain and showed typical birefringence when examined by polarized microscopy. Further workup of the patient revealed generalized amyloidosis with multiorgan involvement by the disease. This case demonstrates that FNA can be a useful technique in the diagnosis of unsuspected amyloidosis.

Aged↗

Mapping of glutaraldehyde-treated bovine pericardium and tissue selection for bioprosthetic heart valves.

Glutaraldehyde-crosslinked bovine pericardium is widely used in bioprosthetic heart valve fabrication. In an attempt to set a scientific basis for more reproducible tissue selection, we produced and analyzed topographical maps of glutaraldehyde-treated bovine pericardium. Whole pericardia were divided into specific anatomical areas and their thickness was measured and mapped on templates. In each area, the suture holding power was determined in both parallel and perpendicular (to the base-apex line) directions; analyses of the tearing patterns in each fragment were used to evaluate predominant fiber orientation, and observations were confirmed by polarized light microscopy. Complete maps were superimposed graphically to aid in the selection of certain areas that would have known fiber orientation, high suture holding power, and suitable thickness. Our results describe regional heterogeneity of bovine pericardial structure and mechanical properties, specifically demonstrating variations in thickness, suture holding power, and collagen fiber orientation. Two areas of choice (representing about 35% of the total) were described as suitable for use in bioprosthetic heart valve fabrication.

Animals↗

Orientation and behavior of epithelial cell muscle processes during Hydra budding.

The reorientation of the muscle processes of the ectodermal epithelial cells in Hydra attenuata has been examined during bud morphogenesis and in grafts inserted at right angles to their original orientation. The muscle processes were observed in histological preparations after staining with Mallory's Triple Stain and, in freshly fixed hydra, with polarization microscopy. Since the bud forms as an outpocketing from the parent, the ectodermal muscle processes, which run longitudinally, must reorient at some stage in order to be in the proper orientation on the bud. During the early stages of bud development, the reorientation of the ectodermal muscle processes is a passive result of the deformation of the parental tissue. Later, active reorientation occurs. The means by which muscle processes are able to reorient was studied further in rectangular grafts placed at right angles to their original orientation. In these grafts, muscle processes become aligned with those of the host in two ways: (1) in groups through the rotation of the whole graft or (2) individually, probably by retracting and then reextending in the new direction.

Animals↗