scholarly journals Body-Surface Compounds in Buckfast and Caucasian Honey Bee Workers (Apis Mellifera)

2014 ◽  
Vol 58 (1) ◽  
pp. 5-15 ◽  
Author(s):  
Aneta Strachecka ◽  
Grzegorz Borsuk ◽  
Jerzy Paleolog ◽  
Krzysztof Olszewski ◽  
Milena Bajda ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Body Surface ◽  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Chemical Compounds ◽  
The Body ◽  
Surface Chemical ◽  
Surface Compounds ◽  
Honey Bee Workers

Abstract Body-surface chemical compounds were studied in 1-day-old nest workers and foragers both in Buckfast and Caucasian bees. The workers of these two age-castes were sampled twice in each of two consecutive years. Body-surface lipids were determined by means of gas chromatography, with a GCQ mass spectrometer. Protein concentrations and activities on the body surface were examined in bee cuticle rinsings obtained from worker bees according to the methods of Lowry, of Anson, and of Lee and Lin. Protease and protease inhibitor activities were determined. Polyacrylamide gel electrophoresis was performed. Caucasian bees, particularly foragers, had more lipids, but Buckfast bees (two age-castes) had more proteins on their body surfaces. A total of 17 alkane types (C17 - C33), 13 alkene types (C21 - C33), 21 esters (C12 - C32), and a phenol (C14) were detected in both races. Alkene C33 was detected only in Caucasian bees. More alkanes, esters, and phenols were found in Caucasian 1-day-old nest workers and foragers than in these age-castes of Buckfast bees. The protein concentration and protease inhibitor activities were lower in Caucasian bees that had higher protease activities. These values corresponded with specific numbers and widths of the electrophoretic bands.

Proteolysis on the body surface of pyrethroid-sensitive and resistant Varroa destructor

2013 ◽  
Vol 58 (1) ◽  
Author(s):  
Aneta Strachecka ◽  
Grzegorz Borsuk ◽  
Krzysztof Olszewski ◽  
Jerzy Paleolog ◽  
Zbigniew Lipiński
Keyword(s):  
Protease Inhibitor ◽  
Proteolytic Activity ◽  
Body Surface ◽  
Serine Proteases ◽  
Protein Concentration ◽  
Proteolytic Enzymes ◽  
Electrophoretic Analysis ◽  
The Body ◽  
Activity Levels

AbstractThe aim of this work was to determine the activity of proteases and protease inhibitors sampled from the body surface of tau-fluvalinate-sensitive and resistant V. destructor. Proteins were isolated from the tau-fluvalinate-sensitive and resistant mites, while mites untreated with tau-fluvalinate constituted the control. Subsequently, the following methodology was applied: protein concentration assay by the Lowry method — as modified by Schacterle and Pollack; assay of proteolytic activity in relation to various substrates (gelatine, haemoglobin, ovoalbumin, albumin, cytochrome C, casein) by the modified Anson method; identification of proteolytic activity in relation to diagnostic inhibitors of proteolytic enzymes (pepstatin A, PMSF, iodoacetamide, o-phenantrolin), using the Lee and Lin method; identification of acidic, neutral and basic protease activities by means of the modified Anson method; electrophoretic analysis of proteins in a polyacrylamide gel for protease detection with the Laemmli method and for protease inhibitor detection with the Felicioli method. The highest value of protein concentration was found in the tau-fluvalinate-sensitive V. destructor, while the highest activity levels of acidic, neutral and alkaline proteases were observed in the tau-fluvalinate-resistant mites. Aspartic, serine, thiolic and metallic proteases were found in the drug-resistant and drug-sensitive Varroa mites. The control samples were found to contain aspartic and serine proteases. In an acidic and alkaline environment, the results revealed a complete loss of inhibitor activities in the in vitro analyses and electrophoresis. Serine protease inhibitor activities (at pH 7.0) were high, especially in the group of tau-fluvalinate-resistant mites.

Normal Mature Human Enamel Protein

1979 ◽  
Vol 58 (2_suppl) ◽  
pp. 986-987 ◽  
Author(s):  
A. Belcourt
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Organic Matrix ◽  
Low Molecular Weight ◽  
Human Enamel ◽  
Weak Density ◽  
Enamel Protein

Pure enamel was prepared using an original microdissection technic. Protein concentration was 375 μg per gram of enamel. Polyacrylamide gel electrophoresis showed a single fast-migrating zone containing a thin double band. Ultracentrifugation studies suggested that the proteins were of low molecular weight or of weak density. Absorption spectra showed a strong absorbance at 260nm. Amino acid analyses yielded a composition of 25% Gly, 13.5% Glu, 11% Ser, 11% Pro, 2% Cys and 2% Hyp. A glucidic content of 15% was estimated and glucose, galactose, mannose and fucose were identified. The organic matrix of enamel seemed to be constituted of two major glycoproteins probably fibrous but different from keratin.

Characterization of the protein Z–dependent protease inhibitor

Blood ◽  
2000 ◽  
Vol 96 (9) ◽  
pp. 3049-3055 ◽  
Author(s):  
Xin Han ◽  
Ryan Fiehler ◽  
George J. Broze
Keyword(s):  
Protease Inhibitor ◽  
Gel Electrophoresis ◽  
Thrombin Generation ◽  
Proteinase Inhibitors ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Factor Xa ◽  
Polyacrylamide Gel ◽  
Protein Z ◽  
Factor Xia

Abstract Protein Z-dependent protease inhibitor (ZPI) is a 72-kd member of the serpin superfamily of proteinase inhibitors that produces rapid inhibition of factor Xa in the presence of protein Z (PZ), procoagulant phospholipids, and Ca++ (t1/2 less than 10 seconds). The rate of factor Xa inhibition by ZPI is reduced more than 1000-fold in the absence of PZ. The factor Xa–ZPI complex is not stable to sodium dodecyl sulfate–polyacrylamide gel electrophoresis, but is detectable by alkaline–polyacrylamide gel electrophoresis. The combination of PZ and ZPI dramatically delays the initiation and reduces the ultimate rate of thrombin generation in mixtures containing prothrombin, factor V, phospholipids, and Ca++. In similar mixtures containing factor Va, however, PZ and ZPI do not inhibit thrombin generation. Thus, the major effect of PZ and ZPI is to dampen the coagulation response prior to the formation of the prothrombinase complex. Besides factor Xa, ZPI also inhibits factor XIa in the absence of PZ, phospholipids, and Ca++. Heparin (0.2 U/mL) enhances the rate (t1/2 = 25 seconds vs 50 seconds) and the extent (99% vs 93% at 30 minutes) of factor XIa inhibition by ZPI. During its inhibitory interaction with factor Xa and factor XIa, ZPI is proteolytically cleaved with the release of a 4.2-kd peptide. The N-terminal amino acid sequence of this peptide (SMPPVIKVDRPF) establishes Y387 as the P1 residue at the reactive center of ZPI. ZPI activity is consumed during the in vitro coagulation of plasma through a proteolytic process that involves the actions of factor Xa with PZ and factor XIa.

scholarly journals Biochemical and isoenzyme analyses of elephant grass, Pennisetum purpureum (Schum) varieties

1995 ◽  
Vol 52 (3) ◽  
pp. 528-533 ◽  
Author(s):  
E.E. Bach ◽  
V.B.G. Alcântara ◽  
P.B. Alcântara ◽  
E.A. Veasey
Keyword(s):  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Plant Age ◽  
Pennisetum Purpureum ◽  
Elephant Grass ◽  
Bud Emergence ◽  
Biochemical Analyses ◽  
Esterase Band

This study characterized seven Pennisetum purpureum varieties, namely v. Anão, Bajra, Cameroon, Guaçu, Roxo, Taiwan A-144 and Uruckwami, through biochemical analyses, including protein, glucose and fructose contents, and polyacrylamide gel electrophoresis using the esterase system, by sampling 30, 60, 90, 120 and 150 day-old leaves. The number of nodes per stem and the percentage of bud emergence were also recorded. Variety Taiwan A-144 presented the highest number of nodes per stem and percentage of emerging buds. Protein concentration decreased gradually after 60 days for all varieties, except for Anão. Variety Guaçu presented the highest level of glucose in 90 day-old plants, whereas Cameroon presented the highest levels at 120 and 150 days. The esterase band patterns changed with plant age for all varieties, showing a tendency to increase the number of bands with time. The best age for discriminating between esterase bands of P. purpureum varieties was at 120 days, when most variation could be observed.

scholarly journals INTERACTIONS BETWEEN SEX-TRANSFORMATION MUTANTS OF DROSOPHILA MELANOGASTER. I. HEMOLYMPH VITELLOGENINS AND GONAD MORPHOLOGY

Genetics ◽  
1981 ◽  
Vol 99 (3-4) ◽  
pp. 429-441
Author(s):  
T Ota ◽  
A Fukunaga ◽  
M Kawabe ◽  
K Oishi
Keyword(s):  
Gel Electrophoresis ◽  
Fat Body ◽  
Biochemical Marker ◽  
Polyacrylamide Gel Electrophoresis ◽  
Body Cavity ◽  
The Body ◽  
The Other ◽  
Sexually Dimorphic ◽  
Gonad Morphology ◽  
Sex Transformation

ABSTRACT In Drosophila, vitellogenins (yolk protein precursors) are synthesized by the female fat body, secreted into the hemolymph and subsequently taken up by the developing oocytes. The male fat body, on the other hand, does not do this even when immature ovaries are transplanted into the body cavity and grow. Thus, the hemolymph vitellogenins serve as an easily detectable sexually dimorphic biochemical marker.——We have examined hemolymph vitellcgenins by SDS polyacrylamide gel electrophoresis in flies carrying various sex-transformation mutants (dsx, tra, tra-2 and tra-20TF) singly and in all possible combinations. Chromosomal females homozygous for tra or tra-2 have no detectable hemolymph vitellogenins, while those homozygous for tra-20TR exhibit appreciable levels of these proteins. Flies homozygous for dsz, bothX / X and X / Y, have hemolymph vitellogenins, although the amount is consistently smaller in the latter. Indeed, X / Y; dsx/dsx is the only genotype in which hemolymph vitellogenins are detected in the X/Y flies. A clear hierarchy of epistasis exists among these sex-transformation mutants when they are examined in various combinations: dsx > tra, tra-2 > tra-20TF. Moreover, an interaction between tra-20TF and tra was seen in these experiments: X / X; tra-20TF/tra-20TF flies show the presence of only a trace of hemolymph vitellogenins when they are made heterozygous for tra. These results, combined with observations on gonad morphology, are discussed with respect to the BAKERa nd RIDGE (1980) hypothesis of sex determination.

scholarly journals Synthesis of lipoprotein lipase in the liver of newborn rats and localization of the enzyme by immunofluorescence

1988 ◽  
Vol 249 (2) ◽  
pp. 549-556 ◽  
Author(s):  
S Vilaró ◽  
M Llobera ◽  
G Bengtsson-Olivecrona ◽  
T Olivecrona
Keyword(s):  
Lipoprotein Lipase ◽  
Gel Electrophoresis ◽  
Time Course ◽  
Polyacrylamide Gel Electrophoresis ◽  
Lipoprotein Metabolism ◽  
The Body ◽  
Newborn Rats ◽  
Adult Rats ◽  
Similar Time ◽  
Extrahepatic Tissues

In newborn rats, lipoprotein lipase (LPL) activity was higher in the liver than in several other tissues, such as heart, diaphragm or lungs, and accounted for about 3% of total LPL activity in the body. There was no significant correlation between LPL activity in liver and in plasma. Thus transport of the enzyme from extrahepatic tissues was probably not the major source of LPL in liver. To study LPL biosynthesis directly, newborn rats were injected intraperitoneally with [35S]methionine, and LPL was isolated by immunoprecipitation and separation by SDS/polyacrylamide-gel electrophoresis. Radioactivity in LPL increased with a similar time course in all tissues studied, including the liver. Substantial synthesis of LPL was also demonstrated in isolated perfused livers from newborn rats, whereas synthesis was low in livers from adult rats. There was strong LPL immunofluorescence in livers from newborn rats, mainly within sinusoids and along the walls of larger vessels. This labelling disappeared after perfusion with heparin, which indicates that much of the enzyme is in contact with blood and can take part in lipoprotein metabolism.

scholarly journals Profile of the body surface proteolytic systém in Apis mellifera quee

2011 ◽  
Vol 56 (No. 1) ◽  
pp. 15-22 ◽  
Author(s):  
A.J. Strachecka ◽  
M.M. Gryzińska ◽  
M. Krauze ◽  
K. Grzywnowicz
Keyword(s):  
Protease Inhibitor ◽  
Honey Bee ◽  
Proteolytic Activity ◽  
Body Surface ◽  
Protease Activity ◽  
Serine Proteases ◽  
Developmental Stages ◽  
Cysteine Proteases ◽  
The Body

The proteolytic system on the body surface of the honey bee has been insufficiently researched. In this study the body surface proteolytic activity was examined in queens at various developmental stages (eggs, larvae, pupae and imagines) in different seasons (spring, summer, autumn, winter). Extracts of the body surface material with water and detergent were used for an in vitro analysis of the proteolytic activity and protease inhibitor level assaying, as well as for an electrophoretic separation of the extracts in polyacrylamide gels. The following methods were used: protein content testing by the Lowry method (modified by Schacterle-Pollack), protease activity testing by the Anson method and protease inhibitor activity testing by the Lee and Lin method. Our studies revealed a high protease activity in an acidic environment (pH = 2.4; the material rinsed with detergent), as well as in neutral (pH = 7) and alkaline (pH = 11.2) environments (the material rinsed with water in both cases). The highest protein concentration values were observed in the imagines from summer. The lowest activities of the proteases and protease inhibitors were determined in the eggs from summer. The highest activities of the acidic, neutral and alkaline proteases were observed in the pupae from spring. The highest number of protease activity bands in PAGE zymography was obtained for the neutral and alkaline activities in the queens for all the seasons. In the queens all the catalytic protease types were present: asparagine and cysteine proteases at pH = 2.4; cysteine proteases and metalloproteases at pH = 7 and serine proteases at pH = 11.2. These results were crucial for the analysis of immunity mechanisms on the body surface of the honey bee.

scholarly journals Gelatin extraction from the indigenous Pangasius catfish bone using pineapple liquid waste

2018 ◽  
Vol 22 (2) ◽  
pp. 86 ◽  
Author(s):  
Yoni Atma ◽  
Hisworo Ramdhani
Keyword(s):  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Liquid Waste ◽  
Environmental Issues ◽  
Fish Bone ◽  
Confirmation Test ◽  
Sds Page ◽  
Pre Treatment

Gelatin extraction from fish bone has traditionally involved hydrogen chloride and/or sodium hydroxide during pre-treatment. However, these chemicals have begun to be abandoned because of their associated safety and environmental issues. Several studies have looked at the use of citric acid as a safer alternative in fish bone gelatin extraction. The aim of this research was to extract gelatin from the bone of Pangasius catfish with pineapple liquid waste. The extraction was performed in two steps: pre-treatment followed by main extraction at various times (24–56 h) and temperatures (45–75°C). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used as a confirmation test and showed a band for gelatin at ~120 kDa. Gelatin yields were calculated as the ratio of weight of dried gelatin to the total weight of fish ossein. The results indicated that pineapple liquid waste can be used for fish bone gelatin extraction. The recommended conditions for extraction of fish bone gelatin using pineapple liquid waste are 56 h of pre-treatment and 5 h of main extraction at a temperature of 75°C. The gelatin yield was 6.12% and the protein concentration 4.00 g/100 g.

Inhibition by chelating agents of the formation of active extracellular proteinase byPseudomonas fluorescens32A

1985 ◽  
Vol 52 (1) ◽  
pp. 91-100 ◽  
Author(s):  
Robin C. Mckeller ◽  
Hilaire Cholette
Keyword(s):  
Gel Electrophoresis ◽  
Protein Concentration ◽  
Chelating Agents ◽  
Polyacrylamide Gel Electrophoresis ◽  
Skim Milk ◽  
Staining Intensity ◽  
Protein Band ◽  
Enzyme Synthesis ◽  
Extracellular Proteinase ◽  
Proteinase Production

SUMMARYThe effect of chelating agents on extracellular proteinase production byPseudomonas fluorescens32A was examined. Increasing concentrations of orthophosphate slightly stimulated growth while inhibiting proteinase synthesis. Fifty percent inhibition was found at 35 and 28 mM-orthophosphate at 5 and 20 °C respectively. Extracellular protein concentration was reduced by 30% when cells were grown with 100 mM-orthophosphate. Polyacrylamide gel electrophoresis of the cell-free supernatants suggested that reduced enzyme synthesis had taken place as evidenced by the decrease in staining intensity of the protein band corresponding to the proteinase. Other phosphate compounds could replace orthophosphate as an inhibitor. Extent of inhibition was related to chain length; polyphosphates with 4–6 or 13–18 phosphorus atoms were the most effective inhibitors. EDTA (0·5 mM) completely inhibited proteinase synthesis. This inhibition could be partly reversed by Ca2+and, to a lesser extent, Mn2+. Proteinase production at 5 °C in skim milk was completely inhibited by phosphate glass (P13–P18). Control experiments showed that loss of activity with chelators was not due to inhibition of preformed enzyme. The results suggest a possible role for polyphosphates in controlling proteinase production in stored milk.

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