Comparative evaluation of the depth of collagen and hyaluronic acid hydrolysis <Emphasis Type="Italic">in vitro</Emphasis> by collagenase and hyaluronidase preparations

The molecular weights of collagen and hyaluronic acid solutions after their incubation with collagenase and hyaluronidase were evaluated by capillary viscosimetry. The results indicate high amylolytic activity of collagenase and the absence of proteolytic activity in hyaluronidase in an in vitro system. Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 146, No. 7, pp. 89–90, July, 2008     

Alterative expression of the collagenase and adhesion molecules in the highly metastatic clones of human colonic cancer cell lines

Human colonic carcinoma cell lines, KM12C, KM12SM and KM12L4, were previously established and their in vivo metastatic potentials have been well evaluated. The highly metastatic cell lines KM12SM and KM12L4 were derived from the parental low metastatic cell line KM12C in vivo. To evaluate the metastatic behavior of these cell lines in vitro, we examined colony formation on monolayers of the pulmonary arterial endothelial (CPAE) cells. On day 4, the highly metastatic cell lines showed an approximately 2-fold increase in number of colonies on CPAE cell monolayers relative to the parental KM12C cell line. To investigate what evidence is correlated with their metastatic and invasive abilities, Northern blot analysis and flow cytometry were performed in all cell lines. According to the results of Northern blot analysis, the levels of matrix metalloproteinase (MMP)-2 and c-met mRNA expression were increased in highly metastatic cell lines as compared with the parental cell line. We also examined the cell-surface expression of several adhesion molecules by flow cytometry. The levels of expression of sialyl Lewisa antigen (sLe(a)) in KM12SM and KM12L4 were twice higher than that in KM12C. However, the levels of expression of E-cadherin in KM12SM and KM12L4 were decreased to half that in KM12C. The alterative expression of the collagenase and adhesion molecules might contribute to their metastatic/invasive abilities of these cell lines both in vivo and in vitro.     

Alterative expression of the collagenase and adhesion molecules in the highly metastatic clones of human colonic cancer cell lines

Human colonic carcinoma cell lines, KM12C, KM12SM and KM12L4, were previously established and their in vivo metastatic potentials have been well evaluated. The highly metastatic cell lines KM12SM and KM12L4 were derived from the parental low metastatic cell line KM12C in vivo. To evaluate the metastatic behavior of these cell lines in vitro, we examined colony formation on monolayers of the pulmonary arterial endothe-lial (CPAE) cells. On day 4, the highly metastatic cell lines showed an approximately 2-fold increase in number of colonies on CPAE cell monolayers relative to the parental KM12C cell line. To investigate what evidence is correlated with their metastatic and invasive abilities, Northern blot analysis and flow cytom-etry were performed in all cell lines. According to the results of Northern blot analysis, the levels of matrix metalloproteinase (MMP)-2 and c-met mRNA expression were increased in highly metastatic cell lines as compared with the parental cell line. We also examined the cell-surface expression of several adhesion mole-cules by flow cytometry. The levels of expression of sialyl Lewis a antigen (sLe a ) in KM12SM and KM12L4 were twice higher than that in KM12C. However, the levels of expression of E-cadherin in KM12SM and KM12L4 were decreased to half that in KM12C. The alterative expression of the collagenase and adhesion molecules might contribute to their metastatic/invasive abilities of these cell lines both in vivo and in vitro. © Rapid Science Ltd.     

Comparative study of the antigenic and immunogenic properties of native and recombinant Marburg virus VP35 proteins]

Antigenic structure of Marburg virus (MBG) VP35 was compared with that of the recombinant VP35 (f35) expressed in a prokaryotic system. For this purpose, a gene encoding the full-length VP35 was cloned in vector pQE31(QIAGEN) and expressed at about 70 mg/liter culture fluid in Escherichia coli JM103 as a recombinant fusion protein f35. BALB/c mice were immunized with soluble f35 or purified inactivated virions of MBG. Antibodies cross-reacting with VP35 and f35 antigens were detected by ELISA and Western Blot analysis in immune sera. Serum from a convalescent after Marburg disease and polyclonal antibodies from animals immunized with MBG recognized f35 and the MBG VP35. VP35 and its recombinant analog induced the production of specific antiviral antibodies in mice, which cross-reacted with the studied antigens. Competitive EIA showed that VP35 and f35 cross-inhibit the antigenic reactivity with polyclonal antibodies of immune sera. Antigenic structure of f35 protein corresponded to antigenic structure of MBG VP35 protein.     

Immunological and physical properties of allergen solutions

Lyophilised birch pollen allergen extracts, reconstituted with different diluents (H₂O, saline, Albumin diluent® (AD)) were investigated to determine whether the allergen activity and quality of the extracts deteriorated by nebulization with different nebulizers (Pari, Wright, and Samdoz). Allergen activity was measured by IgG₄ RAST inhibition technique and allergen quality was analysed by crossed immunoelectrophoresis (CIE). The distribution of particle sizes of aerosols different allergen solutions was determined by a TSI Aerodynamic Particle Sizer. A significant difference (P < 0.05) in allergen activity was found between the AD and H₂O diluents before and after using a Sandoz nebulizer and a Wright nebulizer equipped with a small chamber. This suggested greater allergen activity in AD-diluted solutions, and the pattern was repeated with the other two nebulizers, but was not statistically significant. The samples diluted with saline showed no significant differences in quality after nebulization except for the impacted aerosol in which one of the precipitates was slightly diminished. In the AD-diluted sample one of the precipitates disappeared from the impacted aerosol and from the nebulization chamber after 2 min nebulization. To CIE with rabbit anti-human albumin in an intermediate gel. By this procedure one precipitate was transformed to a precipitate in the intermediate gel, indicating that one or more proteins in the extracts may associate with albumin. No significant difference in output was observed between the nebulizers. The particle size distribution curves for each diluent (saline, AD) were identical for the Wright nebulizer with 99% of the dry particles distributed within 0.5–2.0 μm. Although allergen solutions reconstituted with different diluents were not deteriorated in different nebulizers, it may be that the addition of human albumin to an allergen solution may induce one or more proteins in the extracts to associate with it. This could reduce allergen adsorption over periods longer than this study. The use of different diluents did not change the distribution of particle size generated by the nebulizer tested.     

Regulation of MMP-9 (type IV collagenase) production and invasiveness in gliomas by the extracellular signal-regulated kinase and jun amino-terminal kinase signaling cascades

Our previous studies have shown that MMP-9 levels are significantly elevated during the progression of human gliomas. In the current study, we examined the role of JNK- and ERK-dependent signaling modules in the regulation of MMP-9 production and the invasive behavior of the human glioblastoma cell line SNB19, in which JNK/ERK1 is constitutively activated. SNB19 cells that were transfected with dominant-negative JNK, MEKK, and ERK1 expression vectors showed reduced MMP-9 promoter activity. In addition, conditioned medium collected from SNB19 cells transfected with these expression vectors showed diminished MMP-9 activity in the presence of phorbol myristate acetate, as determined by gelatin zymography. The cotransfection of SNB19 cells with kinase-deficient c-raf also diminished MMP-9 promoter activity. Further, in the presence of a specific inhibitor of MEKK (PD098059), the Matrigel invasion assay showed the invasiveness of dominant-negative SNB19 cells transfected with dominant-negative JNK1 or ERK1 to be remarkably reduced. In conclusion, our studies showed for the first time that MMP-9 production and the invasive behavior of SNB19 cells are regulated by JNK- and ERK-dependent signaling modules and that interfering with either of the pathways reduces invasiveness. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative study of the effects of trophoblastin and polydeoxyribonucleotide preparations on cultured skin fibroblast

We studied the effects of trophoblastin and polydeoxyribonucleotide on the morphology and functional characteristics of fibroblasts in the culture of skin cells. Positive effects of trophoblastin and polydeoxyribonucleotide on fibroblast growth and survival during culturing were demonstrated. Trophoblastin exhibited similar effects on cells of early and late passages. Polydeoxyribonucleotide exhibited a more pronounced effect on cells of late passages, whose growth activity decreased with age. Being a multicomponent preparation, trophoblastin stimulated processes leading to positive dynamic of cell development and vital activity. Polydeoxyribonucleotide had a favorable impact on functioning of basal cell structures at the level of cell nucleus and maintained high level of cells with complete DNA set. __________ Translated from Kletochnye Tehnologii v Biologii i Medicine, No. 3, pp. 173–177, July, 2007     

表面修饰煅烧骨的制备及其理化性质的研究

探讨一种新的仿生骨材料表面修饰煅烧牛松质骨的制备方法,以提高煅烧牛松质骨作为组织工程骨的活性。将制备好的大小一致的煅烧牛松质骨随机分成两组,分别浸泡在配制好的单倍模拟体液(SBF)和1.5倍SBF中。每组材料的浸泡时间均为7、14和21d共3个时间点。浸泡结束干燥后用扫描电镜(SEM)观察材料表面形态并分析材料表面矿化成分。通过比较筛选出效果最理想的表面修饰煅烧牛松质骨材料,研究其孔径、孔隙率、抗压和抗折强度等理化性质,并与未经表面修饰的煅烧骨材料进行比较。研究表明,煅烧骨材料在1.5倍SBF中浸泡14d可以获得最佳表面修饰效果,同时保留了原有煅烧骨材料的基本理化特性。     

Infantile systemic hyalinosis in siblings: Clinical report,biochemical and ultrastructural findings,and review of the literature*

A boy presented at age 3.5 months with joint contractures, restlessness, and pain on handling. His skin was thickened and there were livid‐red macular lesions over bony prominences. Infantile systemic hyalinosis (ISH) was diagnosed, a presumably autosomal recessive, progressive, and painful disorder of as yet unknown pathogenesis. Observation over three years confirmed the diagnosis as typical changes, such as nodules on both ears, pearly papules in the perinasal folds and on the neck, fleshy nodules in the perianal region, and gingival hypertrophy, developed. Skin lesions and painful joint contractures progressed in spite of intense physiotherapy, and at age 3, the child had marked motor disability. The central nervous system (CNS) appeared to be intact and the infant showed normal mental development. Radiologic findings included marked generalized osteopenia, osteolytic erosions in the metaphyses of the long bones, and cortical thinning. Electron microscopy of two skin biopsies demonstrated deposition of floccular amorphous substance that was abundant around, and appeared to originate from, small blood vessels in the dermis, partially interfering with collagen fiber formation. Lysosomal inclusions were not seen. Serum acid hyaluronidase activity was within the normal range, and the synthesis of hyaluronic acid and proteoglycans in cultured skin fibroblasts was similar to that of control cells. A younger sister presented at age two months with painful joint contractures and discrete livid‐red macules over both malleoli, and showed a similar progression of the disorder over the first year of life. The diagnosis of ISH should be considered in infants and children presenting with painful joint contractures and skin lesions. The pathogenesis of this disabling and disfiguring disorder remains unclear. Our data confirm probable autosomal recessive inheritance, and do not support lysosomal storage, hyaluronidase deficiency, or a primary collagen disorder, but indicate that the amorphous material accumulating in the skin and articular soft tissues may originate from the blood circulation. © 2001 Wiley‐Liss, Inc.     

Comparative study of visual inter and intrahemispheric cortico-cortical connections in five native Chilean rodents

Summary Previous studies of the visual cortical organization in the rat and other rodent species have raised the possibility that the visual cortical plan in the rat is common to a large number of species within the order. We have tested this idea by comparing the visual plan in the rat to cortical subdivision schemes obtained from five native Chilean rodent species, including members of the Cricetidae family within the Miomorph group, as well as from the Octodontidae family within the Caviomorph group. Cortical subdivision schemes were inferred from the analysis of the patterns of callosal connections revealed following multiple injections of HRP contralaterally, as well as from ipsilateral cortico-cortical connections observed after small injections of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) into striate cortex. As in the rat, callosal connections in the native rodents concentrate at the border between cytoarchitectonic areas 17 and 18a, and along the borders of discrete, sparsely callosal islands of cortex in lateral peristriate cortex. Furthermore, single injections of WGA-HRP into striate cortex produce multiple, separate fields of labeled cells and terminations in the cortex surrounding area 17. Together, our data supports the idea of a common plan of visual cortical organization among rodents by providing evidence that the visual cortex in the native species is subdivided into multiple visual areas in a manner that resembles the rat cortical plan.     

Comparative morphology and cytology of the olfactory organs in moray eels with remarks on their foraging behavior

Background. This study compares the morphology and cytology of olfactory organs in moray eels (Muraenidae), particularly Siderea grisea and some species of the genera Echidna, Gymnothorax, and Lycodontis, fishes that are top predators in shallow-water marine habitats. Some of the species search visually for food while others search by olfaction. Methods. The morays were collected in the Red Sea; the nasal olfactory organs were dissected and fixed in Bouin's solution for light-microscopy, and 3.5% glutaraldehyde for electron-microscopy studies. Results. In each studied species the olfactory rosettes are elongated structures situated in closed olfactory chambers between anterior tubular inlet nares and slit-form posterior outlet openings. The double row of lamellae constituting these rosettes are round in Siderea and Echidna and elongated in the other species. They are attached at their base to a median raphe and range in number from 20 in the youngest observed Siderea to 168 in Gymnothorax of 1,500 mm total length. As in other teleosts, the lamellae are covered by a ciliated epithelium composed of three types of sensory cells: two of these, ciliated sensory neurons and ciliated supporting cells, differ in detail, length, and thickness of their cilia and intracellular rootlet system; the third type of sensory cells bears microvillae as well as cilia. Proximal, axonal extensions of the ciliated cells cross the basal lamina in bundles and combine to form fila olfactoria from which the two olfactory nerves extend towards the olfactory bulbs. Lateral extensions at the basal parts of these ciliated cells, the so-called spines, cross the membranes of neighboring cells as dendrites, possibly changing part or all of the ciliated epithelium into an olfactory field. The density and number of sensory cells on the lamellae, as well as observed differences in their foraging behavior in nature and captivity, enable the morays to be divided into two groups: one group, in which the lamellae are richly covered with stereocilia, includes species of the genera Siderea and Echidna, that search for food by olfaction, and the second group, which has a great deal less cells with stereocilia and includes the studies species of Gymnothorax and Lycodontis, locates its food visually. © 1995 Wiley-Liss, Inc.     

Preparation and properties of gelatin hydrolysate modified with polysiloxane quaternary ammonium salts

PSiQAEp-GH polymers were synthesized by the reaction of gelatin hydrolysate (GH) and polysiloxane quaternary ammonium salts containing epoxy group (PSiQAEp) with different molecular weight from 3147 to 12996. The results of FTIR, ¹H NMR and ¹³C NMR showed that the reaction occurred between primary amino group of arginine in GH and epoxy of PSiQAEp. The XRD and DSC studies showed that the degree of short-range order of PSiQAEp-GH reduced and its glass transition temperature (Tg) lowered more than 10?°C compared with GH. The determinations of moisture absorption and contact angle (CA) indicated that the hydrophobility of PSiQAEp-GH was better than GH. The tests of inhibitory zone and minimum bactericidal concentration (MBC) illustrated that the PSiQAEp-GHs exhibited excellent antibacterial activity, and the antibacterial activity depended on both the chemical structure of PSiQAEp-GHs and the biological structure of the bacteria.