Localization and immunological characterization of insulin-like peptide(s) in the land snail Otala lactea (Mollusca: Pulmonata)

2000 ◽  
Vol 78 (9) ◽  
pp. 1515-1526 ◽  
Author(s):  
A M Abdraba ◽  
A SM Saleuddin
Keyword(s):  
Digestive Tract ◽  
Ethanol Extract ◽  
Land Snail ◽  
Size Exclusion ◽  
Connective Tissue Layer ◽  
Cerebral Ganglia ◽  
Molecular Masses ◽  
Otala Lactea ◽  
Vitro Analysis

Insulin-like peptides were detected by means of immunological techniques in tissues of the land snail Otala lactea. Insulin-positive cells were detected in all the ganglia except the right parietal ganglion and visceral ganglion. In the digestive tract, insulin-positive cells were found in the muscle and connective tissue layer of the intestine. The amount of insulin-like peptide detected in acid-ethanol extract of brains and digestive tracts from active snails did not differ significantly from that in the corresponding tissues from estivating (dormant) ones. More insulin-like peptide was detected in hemolymph from active snails than in hemolymph from estivating ones. Brains from active snails released insulin-like peptide in vitro. Analysis of the cerebral ganglia or digestive tract extracts by size-exclusion chromatography and insulin RIA revealed more than one fraction with insulin immunoreactivity. Some of these fractions contained material with molecular masses close to those of mammalian insulin or its subunits. Further analysis of the extracts by reverse-phase chromatography also revealed more than one fraction with immunoreactivity. The immunoreactive material from the digestive tract was found to be less hydrophobic than insulin. Western blot analysis of the cerebral ganglia extract revealed more than one band with insulin immunoreactivity. Three of these bands had molecular masses very similar to those of vertebrate insulin, its subunits, and its precursor.

Protein synthesis in vitro by mantle tissue of the land snail Otala lactea: possible insulin-like peptide function

2000 ◽  
Vol 78 (9) ◽  
pp. 1527-1535 ◽  
Author(s):  
A M Abdraba ◽  
A SM Saleuddin
Keyword(s):  
Protein Synthesis ◽  
Cerebral Ganglion ◽  
Land Snail ◽  
Porcine Insulin ◽  
Land Snails ◽  
Size Exclusion ◽  
Cerebral Ganglia ◽  
Molecular Masses ◽  
Otala Lactea

Mantle-collar tissue from adult land snails Otala lactea continuously incorporated labelled amino acids over a 72-h period of incubation in modified culture medium. Acid-saline extract of cerebral ganglia stimulated protein synthesis by the mantle-collar tissue in vitro. This effect was dose-dependent, with the minimum and maximum doses at 0.5 and 2 cerebral ganglion equivalents, respectively. The protein synthesis-stimulating factor(s) from the cerebral ganglia appeared to be proteinaceous and hydrophobic in nature. The cerebral ganglion extract was fractionated by means of a size-exclusion HPLC column. The biological activity was induced by three fractions with estimated molecular masses of 0.82, 1.88, and 4.33 kilodaltons (kDa). Porcine insulin antiserum abolished the activity of the 4.33- and 1.88-kDa fractions but had no significant effect on the activity of the 0.82-kDa fraction. The results suggest the existence in the cerebral ganglia of more than one factor with protein synthesis-stimulating activity. One of these factors could be related to mammalian insulin. Porcine insulin, however, had no significant effect on protein synthesis by the mantle collar in vitro.

Regulation of gene expression in corn (Zea mays L.) by heat shock. II. In vitro analysis of RNAs from heat-shocked seedlings

1983 ◽  
Vol 61 (6) ◽  
pp. 395-403 ◽  
Author(s):  
Chris L. Baszczynski ◽  
David B. Walden ◽  
Burr G. Atkinson
Keyword(s):  
Heat Shock ◽  
Regulation Of Gene Expression ◽  
In Vitro Translation ◽  
Maize Seedlings ◽  
Mrna Pool ◽  
Molecular Masses ◽  
Vitro Analysis ◽  
Shock Proteins ◽  
In Vitro Analysis

Five-day-old maize seedlings subjected to heat shock exhibit a dramatic enhancement in the synthesis of a small group of polypeptides. Isolation of total RNA from control and heat-shocked maize plumules, fractionation of poly(A)+ mRNA by oligo(dT)-cellulose chromatography, and in vitro translations of the RNAs in both the rabbit reticulocyte and the wheat germ systems indicates that there is remarkable fidelity of the mRNA pool obtained from heat-shocked plumules to reproduce in vitro those same polypeptides whose synthesis is greatly elevated in the intact, heat-shocked plumule. Moreover, these heat-shock polypeptides with molecular masses of 108 000, 89 000, 84 000, 73 000, and 18 000 are translated from polyadenylated mRNAs. The absence of a 76 000 dalton heat-shock polypeptide (HSP) and the presence of fewer isoelectric point variants of the 89 000 and 84 000 dalton HSPs among the in vitro translation products suggests that translational and (or) posttranslational regulatory mechanisms might be operative in determining the final spectrum of the maize heat-shock proteins.

scholarly journals Bactericidal, protistocidal, nematodicidal properties and chemical composition of ethanol extract of Punica granatum peel

2019 ◽  
Vol 27 (3) ◽  
pp. 300-306 ◽  
Author(s):  
V. A. Palchykov ◽  
V. V. Zazharskyi ◽  
V. V. Brygadyrenko ◽  
P. O. Davydenko ◽  
O. M. Kulishenko ◽  
...  
Keyword(s):  
Chemical Composition ◽  
Ethanolic Extract ◽  
Ethanol Extract ◽  
Well Being ◽  
Punica Granatum ◽  
Klebsiella Pneumonia ◽  
Leading Role ◽  
Resistant Strains ◽  
Vitro Analysis

We have studied the chemical composition and antibacterial profile of ethanolic extract of Punica granatum L. (Lythraceae) on strains of microorganisms in vitro. Analysis using GC-MS showed 5-hydroxymethylfurfural (36.6%), D-sucrose (23.2%), sorbitol (6.7%), palmitic acid β-monoglyceride (5.6%), 2-furancarboxaldehyde (3.5%) and β-D-glucopyranose (3.3%) as the major components of the title extract. The experiment revealed a positive antibacterial effect of extracts obtained from P. granatum on 14 strains specifically Enterobacteriaceae microorganisms (Escherichia coli, Enterobacter aegorenеs, Proteus vulgaris, Serratia marcescens, Klebsiella pneumonia), Listeriaceae (Listeria ivanovi, L. іnnocua, L. monocytogenes) and yeasts from the family Saccharomycetaceae (Candida albicans). Our study showed that in many cases these extracts more intensively affect multi-resistant strains of microorganisms than macrolide antibiotic azithromycin and is therefore a source of molecules to be exploited in medicine or by the pharmaceutical industry. The investigated extracts of P. granatum can be recommended for further in-depth research against poly-resistant strains of the above-mentioned microorganisms. Effective drugs perform a leading role in providing stable veterinary well-being of livestock and healthcare of the population. The present study showed that the studied plant species more intensively affects multi-resistant strains of microorganisms than sodium salt of azithromycin. Lethal concentration (LC50) of ethanol extract from pomegranate for Paramecium caudatum Ehr. equaled 0.3%. Death of 100% of nematode larvae of Strongyloides papillosus (Ihle) was recorded during 24 h exposition in 20% extract of P. granatum peel.

scholarly journals Uji Aktivitas Antikolesterol Partisi N-Heksana, Metanol Dan Ekstrak Etanol Kulit Jeruk Nipis (Citrus Aurantiifolia) Secara In Vitro

2021 ◽  
Vol 1 ◽  
pp. 403-412
Author(s):  
Iesyi Lutfiyati ◽  
Urmatul Waznah ◽  
S Slamet ◽  
W Wirasti
Keyword(s):  
Ethanol Extract ◽  
Test Sample ◽  
Research Data ◽  
The Body ◽  
Cholesterol Levels ◽  
Lime Peel ◽  
Vitro Analysis ◽  
Citrus Aurantiifolia ◽  
In Vitro Analysis

AbstractCholesterol is an important sterol in human body tissue which belongs to the lipid group but cannot be hydrolyzed. Cholesterol has various uses in the body such as forming steroid hormones in the hormones estrogen and progesterone. However, if cholesterol levels in the blood are too high, it can cause blockage of blood flow which can lead to atherosclerosis. Lime contains secondary metabolites that function to reduce the increase in cholesterol levels in the blood. The purpose of this study was to determine the anticholesterol activity and to determine the EC₅₀ value of partition n-hexane, methanol, ethanol extract of lime peel (Citrus aurantiifolia) in vitro. Analysis of cholesterol activity is known by measuring cholesterol levels in vitro using Lieberman Burchard reagent. The analytical method used UV-Vis spectrophotometry at a wavelength of 665.0 nm with a series of test sample concentrations of 150 µg/ml; 300 µg/ml; 450 µg/ml; 600 µg/ml and 750 µg/ml. The research data shows that the decrease in cholesterol levels is directly proportional to the increase in the concentration in the sample. The EC₅₀ value of the n-hexane partition was 448.76 µg/ml; methanol partition as much as 448.98 µg/ml and ethanol extract as much as 450.18 µg/ml.Keywords: Anticholesterol; in vitro; lime peel; partition. AbstrakKolesterol merupakan sterol yang penting dalam jaringan tubuh manusia yang termasuk pada golongan lipid tetapi tidak dapat terhidrolisis. Kolesterol memiliki berbagai kegunaan dalam tubuh seperti pembentuk hormon-hormon steroid pada hormon esterogen dan progrsteron. Namun, jika kadar kolesterol dalam darah terlalu tinggi maka dapat menyebabkan penyumbatan aliran darah yang dapat mengakibatkan penyakit Aterosklerosis. Jeruk nipis memiliki kandungan metabolit sekunder yang berfungsi untuk mengurangi kenaikan kadar kolesterol dalam darah. Tujuan pada penelitian ini yaitu mengetahui aktivitas antikolesterol dan mengetahui nilai EC₅₀ dari partisi n-heksana, metanol, ekstrak etanol kulit jeruk nipis (Citrus aurantiifolia) secara in vitro. Analisis aktivitas kolesterol diketahui dengan mengukur kadar kolesterol secara in vitro menggunakan pereaksi Lieberman Burchard. Metode analisis menggunakan spektrofotometri UV-Vis pada panjang gelombang 665,0 nm dengan seri konsentrasi sampel uji 150 µg/ml; 300 µg/ml; 450 µg/ml; 600 µg/ml dan 750 µg/ml. Data penelitian menunjukkan penurunan kadar kolesterol berbanding lurus dengan peningkatan konsentrasi pada sampel. Nilai EC₅₀ partisi n-heksana sebanyak 448,76 µg/ml; partisi metanol sebanyak 448,98 µg/ml dan ekstrak etanol sebanyak 450,18 µg/ml. Kata kunci: Antikolesterol; in vitro; kulit jeruk nipis; partisi.

Suppression of Inflammatory Mediators by the Ethanol Extract of Crotalaria verrucosa L. leaf – in vivo and in vitro Analysis

2016 ◽  
Vol 11 (3) ◽  
pp. 1-7
Author(s):  
Kashfia Nawrin ◽  
Mohammad Billah ◽  
Mohammed Jabed ◽  
Mohammad Khalil ◽  
Md. Uddin ◽  
...  
Keyword(s):  
Inflammatory Mediators ◽  
Ethanol Extract ◽  
Vitro Analysis ◽  
In Vitro Analysis

The native structure of the eukaryotic ribosome

1983 ◽  
Vol 41 ◽  
pp. 432-433
Author(s):  
R.A. Milligan ◽  
P.N.T. Unwin
Keyword(s):  
Ribosomal Proteins ◽  
Crystal Form ◽  
Native Structure ◽  
X Ray Diffraction ◽  
Eukaryotic Ribosome ◽  
Vitro Analysis ◽  
In Vitro Analysis ◽  
Resolution Structure ◽  
Stable Unit

A detailed understanding of the mechanism of protein synthesis will ultimately depend on knowledge of the native structure of the ribosome. Towards this end we have investigated the low resolution structure of the eukaryotic ribosome embedded in frozen buffer, making use of a system in which the ribosomes crystallize naturally.The ribosomes in the cells of early chicken embryos form crystalline arrays when the embryos are cooled at 4°C. We have developed methods to isolate the stable unit of these arrays, the ribosome tetramer, and have determined conditions for the growth of two-dimensional crystals in vitro, Analysis of the proteins in the crystals by 2-D gel electrophoresis demonstrates the presence of all ribosomal proteins normally found in polysomes. There are in addition, four proteins which may facilitate crystallization. The crystals are built from two oppositely facing P4 layers and the predominant crystal form, accounting for >80% of the crystals, has the tetragonal space group P4212, X-ray diffraction of crystal pellets demonstrates that crystalline order extends to ~ 60Å.

Crystal structures of plant inorganic pyrophosphatase, an enzyme with a moonlighting autoproteolytic activity

2019 ◽  
Vol 476 (16) ◽  
pp. 2297-2319 ◽  
Author(s):  
Marta Grzechowiak ◽  
Milosz Ruszkowski ◽  
Joanna Sliwiak ◽  
Kamil Szpotkowski ◽  
Michal Sikorski ◽  
...  
Keyword(s):  
Site Directed Mutagenesis ◽  
Size Exclusion ◽  
Inorganic Pyrophosphatase ◽  
Prolonged Storage ◽  
Mitochondrial Targeting ◽  
Cleavage Rate ◽  
Inorganic Pyrophosphate ◽  
Putative Signal Peptide ◽  
Targeting Peptide

Abstract Inorganic pyrophosphatases (PPases, EC 3.6.1.1), which hydrolyze inorganic pyrophosphate to phosphate in the presence of divalent metal cations, play a key role in maintaining phosphorus homeostasis in cells. DNA coding inorganic pyrophosphatases from Arabidopsis thaliana (AtPPA1) and Medicago truncatula (MtPPA1) were cloned into a bacterial expression vector and the proteins were produced in Escherichia coli cells and crystallized. In terms of their subunit fold, AtPPA1 and MtPPA1 are reminiscent of other members of Family I soluble pyrophosphatases from bacteria and yeast. Like their bacterial orthologs, both plant PPases form hexamers, as confirmed in solution by multi-angle light scattering and size-exclusion chromatography. This is in contrast with the fungal counterparts, which are dimeric. Unexpectedly, the crystallized AtPPA1 and MtPPA1 proteins lack ∼30 amino acid residues at their N-termini, as independently confirmed by chemical sequencing. In vitro, self-cleavage of the recombinant proteins is observed after prolonged storage or during crystallization. The cleaved fragment corresponds to a putative signal peptide of mitochondrial targeting, with a predicted cleavage site at Val31–Ala32. Site-directed mutagenesis shows that mutations of the key active site Asp residues dramatically reduce the cleavage rate, which suggests a moonlighting proteolytic activity. Moreover, the discovery of autoproteolytic cleavage of a mitochondrial targeting peptide would change our perception of this signaling process.

1163: Radial Dilation of Ureteral Balloons: A Comparative in-Vitro Analysis

2005 ◽  
Vol 173 (4S) ◽  
pp. 315-316
Author(s):  
Kari Hendlin ◽  
Brynn Lund ◽  
Manoj Monga
Keyword(s):  
Vitro Analysis ◽  
In Vitro Analysis
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