scholarly journals Effects of detergent on the sulphation of chondroitin by cell-free preparations from chick-embryo epiphyseal cartilage

1992 ◽  
Vol 285 (2) ◽  
pp. 577-583 ◽  
Author(s):  
G Sugumaran ◽  
J E Silbert
Keyword(s):  
Chick Embryo ◽  
Supernatant Fraction ◽  
Random Pattern ◽  
Type I ◽  
Type Ii ◽  
Enzyme Preparations ◽  
Enzyme Substrate ◽  
Ionic Detergent ◽  
The Individual ◽  
Triton X

The effects of the non-ionic detergent Triton X-100 on 6-sulphation of two species of endogenous nascent proteochondroitin by a chick-embryo cartilage microsomal system was examined. Sulphation of the larger (Type I) species with adenosine 3′-phosphate 5′-phosphosulphate was slightly diminished when Triton X-100 was present, whereas sulphation of the smaller (Type II) species was slightly enhanced. An ordered rather than random pattern of sulphation was obtained for the smaller proteoglycan, but with a considerably lower degree of sulphation than that of the larger proteochondroitin. These differences were consistent with other differences between these two species as described previously. Sulphation of exogenous [14C]chondroitin and exogenous proteo[3H]chondroitin by the microsomal system with Triton X-100 present produced ordered rather than random sulphation patterns. When a 100,000 g supernatant fraction was utilized for sulphation of [14C]chondroitin or proteo[3H]chondroitin, Triton X-100 was not needed, and ordered sulphation was still obtained. When hexasaccharide was used, sulphation of multiple N-acetylgalactosamine residues of the individual hexasaccharides resulted. This was relatively independent of Triton X-100 or the concentration of the hexasaccharide acceptors. With soluble enzyme, sulphation of multiple N-acetylgalactosamine residues on the individual hexasaccharide molecules was even greater, so that tri-sulphated products were found. This suggests that ordered rather than random sulphation of chondroitin with these enzyme preparations is due to enzyme-substrate interaction rather than to membrane organization.

scholarly journals Formation of two species of nascent proteochondroitin in separate loci of a microsomal preparation from chick-embryo epiphyseal cartilage

1991 ◽  
Vol 277 (3) ◽  
pp. 787-793 ◽  
Author(s):  
G Sugumaran ◽  
J E Silbert
Keyword(s):  
Chick Embryo ◽  
Type I ◽  
Type Ii ◽  
Microsomal Preparation ◽  
Intact Membrane ◽  
Ionic Detergent ◽  
Before And After ◽  
Microsomal Membranes ◽  
Triton X ◽  
Relationship Of

The potential relationship of an intact membrane organization to the synthesis of chondroitin was examined before and after modification of a chick-embryo cartilage microsomal system with the non-ionic detergent Triton X-100. Incubations with labelled UDP-GlcA and UDP-GalNAc indicated that Triton X-100 had little effect on the amount of chondroitin synthesized to form one species of large proteochondroitin (Type I). However, Triton X-100 had a marked stimulatory effect on the formation of another smaller species of proteochondroitin (Type II). Presence of this detergent during chondroitin polymerization also resulted in chains that were slightly smaller. Neither of the two proteochondroitin species were collagenase-sensitive, nor did they contain dermatan-like regions. Thus in these respects they were unlike the small proteochondroitins (PG-Lb or PG-Lt) that have been found in chick-embryo cartilage. They also differed greatly in size from these small proteoglycans as well as from the large aggregatable proteochondroitin (PG-H) from the same source. Synthesis of the larger (Type I) proteochondroitin species was not affected by prior treatment of the microsomes with chondroitin ABC lyase at concentrations sufficient for elimination of synthesis of most of the smaller (Type II) proteochondroitin species. Use of chondroitin ABC lyase subsequent to synthesis of the chondroitin also resulted in preferential degradation of the smaller species. Thus there were differences in formation and limitation in access of the chondroitin ABC lyase to the two species, consistent with other differences described previously. These results indicate that there are separate loci within the microsomal membranes for synthesis of the two species.

scholarly journals Catalytic properties of lysyl hydroxylase from cells synthesizing genetically different collagen types

1982 ◽  
Vol 201 (1) ◽  
pp. 215-219 ◽  
Author(s):  
U Puistola
Keyword(s):  
Chick Embryo ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Type I ◽  
Type Ii ◽  
Collagen Types ◽  
Enzyme Preparations ◽  
Lysyl Hydroxylase ◽  
Type Iv ◽  
Sarcoma Cells

Crude preparations of lysyl hydroxylase were extracted from chick-embryo tendons synthesizing exclusively type I collagen, chick-embryo sterna synthesizing exclusively type II collagen and HT-1080 sarcoma cells synthesizing exclusively type IV collagen. No differences were found in the Km values for Fe2+, 2-oxoglutarate and ascorbate between these three enzymes preparations. Similarly no differences were found in the Km values for type I and type II protocollagens and the rate at which type IV protocollagen is hydroxylated between these enzyme preparations. The extent to which type I protocollagen could be hydroxylated by the three enzymes was likewise identical. These data strongly argue against the existence of collagen-type-specific lysyl hydroxylase isoenzymes.

Correlation studies of individual variation in susceptibility to various components of HPNS in mice

1981 ◽  
Vol 50 (2) ◽  
pp. 272-278 ◽  
Author(s):  
R. W. Brauer ◽  
R. W. Beaver ◽  
H. W. Gillen
Keyword(s):  
Personnel Selection ◽  
Compression Rate ◽  
Type I ◽  
Type Ii ◽  
Correlation Studies ◽  
Individual Susceptibility ◽  
Selection Strategies ◽  
Stable Characteristic ◽  
The Individual ◽  
Rate Correlation

Individual convulsion threshold pressures were determined in mice exposed successively to type I and type II convulsions of the high-pressure neurological syndrome (HPNS), as well as in others exposed, in successive compressions, to type I convulsions under diverse conditions of replication of compression rate. Correlation analyses of the results showed the following degrees of correlation of individual convulsion-threshold pressures: type I with type II-negligible (r2 less than equal to 0.2); type I with type I at the same compression rate-closely correlated (r2 greater than or equal to 0.8); type I with type I at a different compression rate-negligible (r2 less than or equal to 0.2). Individual susceptibility to HPNS (type I) convulsions thus is a stable characteristic of individual seizures vary independently of one another. Likewise, the magnitude of the individual compression rate effect varies independently of intrinsic individual susceptibility to type I HPNS seizures. The results support the view that the HPNS is a composite entity, define constraints on personnel selection, and provide a basis for estimating the efficacy of various selection strategies.

Expansion of the Mandibular Arch in Children during the Mixed Dentition Period - A Clinical Study

2006 ◽  
Vol 30 (4) ◽  
pp. 329-332 ◽  
Author(s):  
Kenshi Maki ◽  
Yasuhiro Sorada ◽  
Toshihiro Ansai ◽  
Takahiro Nishioka ◽  
Raymond Braham ◽  
...  
Keyword(s):  
Mixed Dentition ◽  
Dental Arch ◽  
Type I ◽  
Type Ii ◽  
Tooth Replacement ◽  
Lateral Expansion ◽  
Incisor Tooth ◽  
Mandibular Arch ◽  
The Individual ◽  
Apical Base

A review of the dental literature revealed relatively few studies on the expansion of the mandibular dental arch. The present study attempted expansion of the mandibular arch using a Bihelix appliance. The subjects were 16 children, exhibiting crowding, age ranges from 7 to 11 years. The mandible was expanded 2.0 mm every 3 months. Significant expansion, not only of the individual tooth inter-arch dimensions but also of the overall arch length, was obtained during the period of incisor tooth replacement. The mode of expansion was classified as follows: Type I, those, which showed no effect on the apical base; Type II, those which showed no consistency of the measurement lines. In this study, 6 of16 cases were classified as type I and 10 cases as type II. Expansion was continued over a period of 1.5 to 3 years. We concluded that considerable lateral expansion of the mandibular arch is possible using the Bihelix appliance. It is suggested that this might contribute greatly to non-extraction orthodontic treatment. Further studies are recommended.

scholarly journals High-yield Production of Recombinant Platelet Factor 4 Protein by Harnessing and Honing the Gram-negative Bacterial Secretory Apparatus

2019 ◽  
Author(s):  
Saeed Ataei ◽  
Mohammad Naser Taheri ◽  
Fatemeh Taheri ◽  
Farahnaz Zare ◽  
Niloofar Amirian ◽  
...  
Keyword(s):  
Light Scattering ◽  
Secondary Structure ◽  
Protein Secretion ◽  
Type I ◽  
Type Ii ◽  
Platelet Factor ◽  
Gram Negative ◽  
Extracellular Milieu ◽  
Secretory System ◽  
Triton X

AbstractBackgroundPlatelet factor 4 is a cytokine released into the bloodstream by activated platelets and plays a pivotal role in heparin-induced thrombocytopenia etiology and diagnosis. Therefore, a sustainable source of recombinant PF4 with structural and functional similarity to its native form is urgently needed to be used in diagnostic procedures.To this end, a three-in-one primary construct was designed and custom synthesized based on the pET26b backbone from which three secondary constructs could be derived each capable of employing either type I, type II secretory or cytoplasmic pathways. Protein expression and secretion were performed in Escherichia coli BL-21 (DE3) and were confirmed by SDS-PAGE and Western blotting. To further enhance protein secretion, the effect of several controllable factors including IPTG, Triton X-100, Sucrose, and Glycine were individually investigated at first. In the next step, according to fractional factorial approach, the synergistic effect of IPTG, Triton X-100, and Glycine on secretion was further investigated. To ascertain the structure and function of the secreted recombinant proteins, Dynamic light scattering was utilized and confirmed rPF4 tetramerization and heparin-mediated ultra-large complex formation. Moreover, Raman spectroscopy was exploited to determine the rPF4 secondary structure.ResultsType II secretory pathway was proven to be superior over type I in case of rPF4 secretion into the extracellular milieu. Protein secretion mediated by Type II was enhanced to approximately more than 700 μg/ml. Large quantities of native rPF4 up to 20 mg was purified upon a minor scale up to 40 ml of culture medium. Dynamic light scattering unveiled native rPF4 quaternary structure revealing the formation of tetramers having an average size of 10 nm and formation of larger complexes of approximately 100-1200 nm in size following heparin supplementation, implying proper protein folding, tetramerization, and antigenicity. Analysis of the Zeta potential on approximately 600 μg/ml of rPF4 revealed a 98 mV positive charge which further confirms protein folding. Moreover, rPF4 secondary structure was determined to be 43.5% Random coil, 32.5% β-sheet, 18.6 % α-helix and 4.9 % Turn, which is in perfect agreement with the native structure.Conclusionour results indicate that the gram-negative type II bacterial secretory system holds a great promise to be employed as a reliable protein production strategy with favorable industrial application. However, further efforts are required to realize the full potential of secretory pathways regarding their application to proteins with distinct characteristics.Abstract FigureGraphical Abstract.rPF4 secretion mediated by type 2 secretory system. The pelB signal sequence directs protein export into the extracellular milieu through the SecYEG translocon complex in a process assisted by SecB chaperone. A) Indicates protein secretion before supplementation with additives and B) indicates secretion in the presence of additives.

scholarly journals STUDIES ON IMMUNOLOGICAL RELATIONSHIPS AMONG THE PNEUMOCOCCI

1929 ◽  
Vol 49 (2) ◽  
pp. 183-193 ◽  
Author(s):  
John Y. Sugg ◽  
James M. Neill
Keyword(s):  
Horse Serum ◽  
Rabbit Serum ◽  
Type I ◽  
Type Ii ◽  
Immunological Relationship ◽  
The Individual ◽  
Relationship Of ◽  
Convincing Data

The paper reports evidence of an immunological relationship between one variety of Saccharomyces ceremsise and the Type II variety of Diplococcus pneumonix (Pneumococcus). The most convincing data consisted of the reactions of the Type II bacteria with potent antiyeast serum which agglutinated, and protected mice against these pneumococci as well as the average antiserum obtained by immunization of rabbits with Type II bacteria themselves. The reactivity of the antiyeast serum is strictly specific to the Type II variety of Pneumococcus in the sense that it is entirely devoid of antibodies reactive with Type I or III. The results of absorption experiments with both the antiyeast (rabbit) serum and the anti-Type II (horse) serum were the same as those usually obtained in analogous experiments with immunologically related, but not identical, kinds of bacteria. The immunological relationship of the yeast and the Type II pneumococcus is apparently based upon S-anti-S reactions. It represents an example of heterogenetic specificity which is of particular interest because of the wide genetic separation of the pathogenic schizomycete and the saprophytic ascomycete. Data on the individual irregularity in the yeast-agglutinating capacity of serum from non-immunized or "normal" rabbits are presented as experimental facts.

scholarly journals Characterization of the protein kinase activities of human platelet supernatant and particulate fractions

1984 ◽  
Vol 218 (2) ◽  
pp. 285-294 ◽  
Author(s):  
S E Salama ◽  
R J Haslam
Keyword(s):  
Protein Kinase ◽  
Cyclic Amp ◽  
Soluble Fraction ◽  
Deae Cellulose ◽  
Type I ◽  
Type Ii ◽  
Dependent Protein Kinase ◽  
Regulatory Subunits ◽  
Dependent Protein ◽  
Triton X

After human platelets were lysed by freezing and thawing in the presence of EDTA, about 35% of the total cyclic AMP-dependent protein kinase activity was specifically associated with the particulate fraction. In contrast, Ca2+-activated phospholipid-dependent protein kinase was found exclusively in the soluble fraction. Photoaffinity labelling of the regulatory subunits of cyclic AMP-dependent protein kinase with 8-azido-cyclic [32P]AMP indicated that platelet lysate contained a 4-fold excess of 49 000-Da RI subunits over 55 000-Da RII subunits. The RI and RII subunits were found almost entirely in the particulate and soluble fractions respectively. Chromatography of the soluble fraction on DEAE-cellulose demonstrated a single peak of cyclic AMP-dependent activity with the elution characteristics and regulatory subunits characteristic of the type-II enzyme. A major enzyme peak containing Ca2+-activated phospholipid-dependent protein kinase was eluted before the type-II enzyme, but no type-I cyclic AMP-dependent activity was normally observed in the soluble fraction. The particulate cyclic AMP-dependent protein kinase and associated RI subunits were solubilized by buffers containing 0.1 or 0.5% (w/v) Triton X-100, but not by extraction with 0.5 M-NaCl, indicating that this enzyme is firmly membrane-bound, either as an integral membrane protein or via an anchor protein. DEAE-cellulose chromatography of the Triton X-100 extracts demonstrated the presence of both type-I cyclic AMP-dependent holoenzyme and free RI subunits. These results show that platelets contain three main protein kinase activities detectable with histone substrates, namely a membrane-bound type-I cyclic AMP-dependent enzyme, a soluble type-II cyclic AMP-dependent enzyme and Ca2+-activated phospholipid-dependent protein kinase, which was soluble in lysates containing EDTA.

scholarly journals 230 years of AGN monitoring: Frequency of cloud occultation events in AGN & constraints for clumpy torus models

2013 ◽  
Vol 9 (S304) ◽  
pp. 265-265 ◽  
Author(s):  
Mirko Krumpe ◽  
A. Markowitz ◽  
R. Nikutta
Keyword(s):  
Homogeneous Medium ◽  
Selection Function ◽  
Base Line ◽  
Type I ◽  
Type Ii ◽  
X Ray ◽  
Cloud Properties ◽  
Monitoring Frequency ◽  
Significant Difference ◽  
The Individual

AbstractWe systematically search for discrete absorption events in the vast archive of the Rossi X-ray Timing Explorer. This includes dozens of nearby type I and Compton-thin type II AGN and covers timescales from days to over a decade for individual objects. We are sensitive to discrete absorption events due to clouds of full-covering, neutral or mildly-ionized gas with columns 1022−25 cm−2 transiting the line of sight. We detect 13 eclipse events in 8 objects, roughly tripling the number of previously published events from this archive. Despite sensitivity to events with NH up to 1024−25 cm−2, we measured no Compton-thick eclipses in our sample. Peak column densities span 2.5–19 × 1022 cm−2. Event durations span hours to months. We infer the clouds distances from the black hole, assuming Keplerian motion, to span 0.2–80 × 104 Schwarzschild radii. We find no statistically significant difference between the individual cloud properties of type I and II objects. The presence of eclipses in both type Is and IIs argues against sharp-edged cloud distributions. The type II AGN show a level of “base-line” X-ray absorption that is consistent with being constant over timescales from 0.6 to 8.4 yr. This can either be explained by a homogeneous medium, or by X-ray-absorbing clouds that each have NH ≪ 1022 cm−2. Considering the “selection function” of the monitoring, we derive the probability of cloud occultation events. Finally, we derive the first X-ray statistical constraints for clumpy-torus models.

scholarly journals Innovative activity of teachers - myth or reality?

2019 ◽  
Vol 21 (5) ◽  
pp. 9-41 ◽  
Author(s):  
A. P. Usol’tsev ◽  
E. P. Antipova
Keyword(s):  
Educational System ◽  
Professional Community ◽  
Innovative Activity ◽  
Practical Significance ◽  
Design Activity ◽  
Type I ◽  
Type Ii ◽  
Creative Activity ◽  
Type Iii ◽  
The Individual

Introduction. The innovative activity of teachers is usually described as the creative activity of a sole inventor and the activity of mass teachers, who follow prepared algorithms in the framework of large-scale innovative projects. It is obvious that these types of activity differ considerably. More accurate structuring of innovative activity of the teacher will allow experts to describe more precisely objective essence of innovative processes and to find out more effective means of their stimulation both at the individual level of the teacher and at the level of a national educational system.The aim of the publication was to discuss the questions of what is innovative activity of the teacher; whether innovative activity really exists in mass practice; how innovative activity can be stimulated.Methodology and research methods. The research was based on the systematic approach. In the course of the research, the analysis of scientific and methodological literature and the survey among the teachers aimed at identifying the characteristics of their innovative activity were employed.Results and scientific novelty. The whole cycle of promoting the use of innovations into practice is thoroughly examined: from subjectivity to objectivity; from creativity to reproductive activity; from objectively new result to the product of a sample; from resistance of environment against innovation to stimulation and coercion to its realisation. Characteristics of innovative activity of the teacher are determined and described by the level of distribution of the forthcoming innovation and the attitudes of professional community towards it. It is proposed and proved that the innovation activities of the teachers should be separated into three types: (type I) creative activity of the individual aimed at development of means, methods and technologies to obtain objectively new results; (type II) activities of the teachers focused on the implementation of ready tested innovations, but new for teacher-friendly professional environment, (start-up); (type III) innovative activity of the teachers as a routine operation in the innovation system as its element. The activity of the type I is not widespread; such activity cannot and should not be mass - not everyone can be engaged in it. Design activity corresponds to the type II; type III - activity of performers with the total development of innovation becoming the accepted norm. Mass introduction of a novelty is an innovative process, but not at the level of a certain subject, since a teacher, who acts (often involuntarily) on well-developed samples and algorithms, is not an innovator.Practical significance. The dependence of efficiency of pedagogical innovations on the model of state policy in education is demonstrated. The reasons of the development gap of the Russian educational system in the field of innovations from the systems of other countries are disclosed. The conditions specific to each type of a teacher’s innovative activity, which need to be created for successful and more rapid implementation of progressive innovations, are allocated.

Postburn Neck Contracture: Principles of Reconstruction and a Treatment Algorithm

2018 ◽  
Vol 34 (07) ◽  
pp. 514-521 ◽  
Author(s):  
Yashan Gao ◽  
Haizhou Li ◽  
Bin Gu ◽  
Feng Xie ◽  
Hainan Zhu ◽  
...  
Keyword(s):  
Treatment Algorithm ◽  
Random Pattern ◽  
Type I ◽  
Type Ii ◽  
Pedicled Flaps ◽  
Type Iii ◽  
Neck Reconstruction ◽  
Normal Neck ◽  
Neck Function ◽  
Location Type

Background Various techniques have been developed for postburn neck reconstruction, but a treatment algorithm is needed. Methods We retrospectively reviewed all patients treated for postburn neck contracture at our institution between February 2008 and December 2015. Necks were divided into one anterior subunit and two lateral subunits marked by the sternocleidomastoid muscle. Deformities were categorized into three types according to their size and location. Type I deformities involve less than one subunit, type II deformities involve at least one subunit but less than two subunits, and type III deformities affect two or more subunits. Type II deformities were further divided into type IIa deformities, which mainly involve the anterior region, and type IIb deformities, which mainly involve the lateral region. Results Local random pattern flaps were constructed for type I deformities. Pedicled flaps from the anterior chest and supraclavicular areas were preferred for type IIa deformities, and pedicled flaps from the back were preferred for type IIb deformities. Pedicled flaps from other areas were the second choice for type II deformities, followed by free and prefabricated flaps. For type III deformities, bipedicled flaps were usually required. At a follow-up of at least 12 months, all patients showed near-normal neck function, and aesthetic features were significantly improved. Conclusion The proposed classification and treatment algorithm for postburn neck reconstruction may help achieve satisfactory outcomes.

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