Sugar-based tertiary amino gemini surfactants with a vesicle-to-micelle transition in the endosomal pH range mediate efficient transfection in vitro

Neutrophils produce hypochlorous acid (HOCl) as well as other reactive oxygen species as part of a natural innate immune response in the human body; however, excessive levels of HOCl can ultimately be detrimental to health. Recent reports suggest that betacyanin plant pigments can act as potent scavengers of inflammatory factors and are notably effective against HOCl. Comparison of the in vitro anti-hypochlorite activities of a novel betalain-rich red beetroot (Beta vulgaris L.) extract with its pure betalainic pigments revealed that the extract had the highest anti-hypochlorite activity, far exceeding the activity of all of the betalainic derivatives and selected reference antioxidants. This suggests that it may be an important food-based candidate for management of inflammatory conditions induced by excessive HOCl production. Among all pigments studied, betanidin exhibited the highest activity across the pH range.

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Metalloproteases and egg-hatching in Pediculus humanus, the body (clothes) louse of humans (Phthiraptera: Insecta)

Parasitology ◽

10.1017/s0031182007003587 ◽

2007 ◽

Vol 135 (1) ◽

pp. 125-130 ◽

Cited By ~ 9

Author(s):

V. M. BOWLES ◽

A. R. YOUNG ◽

S. C. BARKER

Keyword(s):

Protease Activity ◽

The Body ◽

Egg Hatching ◽

Egg Hatch ◽

Pediculus Humanus ◽

Sds Page ◽

Metalloprotease Inhibitors ◽

Egg Shell ◽

Ph Range

SUMMARYTo investigate the biochemical components of egg-hatch in the body louse, Pediculus humanus, egg-shell-washings (ESW) were collected during the first 2 h post-hatching and analysed by gelatin SDS-PAGE. These ESW contained proteases with molecular mass in the range of 25–100 kDa; the most abundant proteases were ~25 kDa. The 3 main regions of protease activity in the one-dimensional gelatin SDS-PAGE gels resolved to at least 23 distinct regions of protease activity when analysed by two-dimensional gelatin SDS-PAGE, with iso-electric points spread over the entire 3 to 10 pH range. Mechanistic characterization indicated that the ESW contained proteases of the metallo-class, inhibited by both 1,10-phenanthroline and EDTA. Several protease inhibitors were tested for their ability to inhibit louse egg-hatch in vitro. The metalloprotease inhibitor 1,10-phenanthroline and the aminopeptidase inhibitor bestatin significantly inhibited (P<0·05) louse egg-hatch (100% and 58%, respectively). The presence of metalloproteases at the time of egg-hatch and the inhibition of egg-hatch in P. humanus by metalloprotease inhibitors suggests a crucial role for these proteases in the hatching of this medically important parasite.

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Isolated flagellar apparatus of Chlamydomonas: characterization of forward swimming and alteration of waveform and reversal of motion by calcium ions in vitro

Journal of Cell Science ◽

10.1242/jcs.33.1.235 ◽

1978 ◽

Vol 33 (1) ◽

pp. 235-253 ◽

Cited By ~ 1

Author(s):

J.S. Hyams ◽

G.G. Borisy

Keyword(s):

Calcium Ions ◽

Beat Frequency ◽

Flagellar Apparatus ◽

Adenosine Monophosphate ◽

Adenosine Diphosphate ◽

Living Cells ◽

Exogenous Calcium ◽

Ph Range ◽

Relationship Of

The control of flagellar activity in the biflagellate green alga, Chlamydomonas reinhardtii was investigated by the in vitro reactivation of the isolated flagellar apparatus (the 2 flagella attached to their respective basal bodies plus accessory structures). The waveform and beat frequency of the isolated apparatus in the presence of 1 mM adenosine triphophate (ATP) were comparable to those recorded for living cells. Equimolar concentrations of adenosine diphosphate (ADP) could be substituted for ATP with little change in beat frequency and no apparent change in waveform, suggesting that the latter is converted to ATP by axonemal adenylate kinase. No reactivation occurred in adenosine monophosphate (AMP). But frequencies in cytidine, guanosine and uridine triphosphates (CTP, GTP and UTP) were approximately 10% that obtained in ATP. Reactivation was optimal over a broad pH range between pH 6.4 and pH 8.9 in both APT and ADP. Isolated flagellar apparatus could be induced to change from forward to reverse motion in vitro by manipulation of exogenous calcium ions. The 2 types of motion were directly comparable to recorded responses of living cells. Forward swimming occurred at levels of calcium below 10(−6)M, the isolated apparatus changing to backward motion above this level. Motility was inhibited at concentrations above 10(−3)M. The threshold for reversal of motion by calcium was lowered to 10(−7)M when the flagellar membranes were solubilized with detergent, indicating that the flagellar membranes are involved in the regulaion of the level of calcium within the axoneme. The reversal of motion by calcium was itself freely reversible. The relationship of these observations to the known tactic responses of Chlamydomonas is discussed.

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pH-induced hysteretic properties of phosphofructokinase purified from rat myocardium

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1986.250.3.r505 ◽

1986 ◽

Vol 250 (3) ◽

pp. R505-R511 ◽

Cited By ~ 2

Author(s):

S. C. Hand ◽

J. F. Carpenter

Keyword(s):

Catalytic Activity ◽

Metabolic Regulation ◽

Inverse Correlation ◽

Rat Myocardium ◽

Solution Ph ◽

Reversible Inactivation ◽

Ph Range ◽

Hysteretic Loss ◽

Fructose 1,6 Bisphosphate

Phosphofructokinase (PFK) purified from the rat myocardium is reversibly inactivated under a pH regime approximating that reported for ischemic hearts. At pH 6.5 and 37 degrees C, the enzyme displays a hysteretic loss of activity during 60-min incubations, declining to 48% of control (pH 7.1, 37 degrees C) values. Citric acid increases the degree of inactivation (28% of control), whereas fructose 1,6-bisphosphate reduces the decline in activity. Simultaneous measurements of 90 decreases light scattering and catalytic activity suggest the inactivation is temporally linked to dissociation of active tetrameric enzyme into an inactive form of lower molecular weight. Fluorescence enhancement of the extrinsic probe sodium mansate, which binds preferentially to dimeric PFK, indicates that the equilibrium dimer concentration (cp1 infinity) increases as pH is lowered. This increase in cp1 infinity exhibits a strong inverse correlation (r = 0.984) with catalytic activity across the pH range of 8.0 to 6.5. Returning solution pH to 7.0 or above promotes a time-dependent reactivation and repolymerization of PFK. The rate of reactivation is increased at higher enzyme concentrations and in the presence of trimethylamine-N-oxide, a nitrogenous osmolyte noted for its ability to promote protein aggregation reactions. Thus these results demonstrate the capacity of rat heart PFK to undergo reversible inactivation and dissociation in vitro and represent the first phase of a two-part study testing the hypothesis that these pH-induced hysteretic processes are operative in the ischemic myocardium. The data are evaluated in terms of the potential roles of hysteretic enzymes in metabolic regulation.

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Evaluation of in vitro antagonistic activity of fungi from peatlands against Ganoderma species under acidic condition

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d210709 ◽

2020 ◽

Vol 21 (7) ◽

Author(s):

SUPRIYANTO ◽

PURWANTO ◽

S.H. POROMARTO ◽

SUPYANI

Keyword(s):

Biological Control ◽

Antagonistic Activity ◽

Biological Control Agents ◽

Antagonistic Fungi ◽

High Incidence ◽

Acidic Conditions ◽

Mineral Soils ◽

Ph Range ◽

Biological Methods

Abstract. Supriyanto, Purwanto, Poromarto SH, Supyani. 2020. Evaluation of in vitro antagonistic activity of fungi from peatlands against Ganoderma species under acidic conditions. Biodiversitas 21: 2935-2945. The use of peatlands is a significant contributor to the world’s palm oil production. A serious problem of oil palm plantations in peatlands is the high incidence of basal stem rot (BSR) disease caused by Ganoderma, which has a higher attack rate than on mineral soils. There is no effective way to control Ganoderma in peatlands. At present, the effort for the same focuses on environment-friendly biological methods; however, this is constrained by the unavailability of appropriate biological agents for peatlands. The development of biological control agents for peatlands is hampered by limited data on biological control of Ganoderma in peatlands. This research was conducted to evaluate the in vitro antagonistic activity of fungi isolated from a peatland in acidic pH conditions. Twenty-seven Ganoderma-antagonistic fungi from peatland were evaluated for their activity and their ability to antagonism in vitro within a pH range of 2-7. The results show that most antagonistic fungi from peatland, based on biomass weight, the sporulation ability, and germination of conidium, were able to grow optimally at pH 3.0-4.0, indicating that most of the Ganoderma-antagonistic fungi from peatland can be used as biological control agents for BSR on oil palms in peatlands.

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Effects of androgens on bioactivity and immunoreactivity of pituitary FSH in GnRH antagonist-treated male rats

Acta Endocrinologica ◽

10.1530/acta.0.1220168 ◽

1990 ◽

Vol 122 (2) ◽

pp. 168-174 ◽

Cited By ~ 12

Author(s):

Om P. Sharma ◽

Shafiq A. Khan ◽

Gerhard F. Weinbauer ◽

Mohammed Arslan ◽

Eberhard Nieschlag

Keyword(s):

Isoelectric Focusing ◽

Gnrh Antagonist ◽

Molecular Composition ◽

Male Rats ◽

Sprague Dawley ◽

Sprague Dawley Rats ◽

Combined Administration ◽

Ph Range ◽

Fsh Bioactivity

Abstract The effects of androgens on the bioactivity and molecular composition of pituitary FSH were examined in intact and GnRH antagonist-suppressed male rats. Eight groups of adult Sprague-Dawley rats were subjected to the following treatments: antagonist (75 μg/day by osmotic minipumps; sc), testosterone-filled Silastic implants (3×5 cm, sc), dihydrotestosterone-filled Silastic implants (3×5 cm, sc), E2 benzoate (15 μg/day, sc), and combined administration of antagonist with either steroid for 3 weeks. At the end of the treatment period, pituitaries were dissected out and homogenised. FSH content was determined in the pituitary extracts by an in vitro bioassay and a radioimmunoassay. Individual pituitary extracts from rats treated with vehicle, testosterone and testosterone + antagonist were subjected to isoelectric-focusing on sucrose density gradients performed in the pH range from 3.5 to 7.0. Individual isoelectric-focusing fractions (100-120) were analysed for bioactive and immunoreactive FSH. Treatment with antagonist, E2 or antagonist + E2 caused a significant decrease in pituitary FSH, whereas testosterone and dihydrotesterone alone or in combination with antagonist prevented the decrease in pituitary FSH. The effects of all treatments on both bioactive and immunoreactive FSH were similar. Testosterone treatment not only maintained FSH synthesis but also altered the molecular composition of pituitary FSH. Following treatment with testosterone there was a shift of maximal FSH bioactivity to the more acidic pH range. On the other hand, less bioactivity was recovered than corresponding immunoreactivity in the higher pH region, resulting in significantly reduced ratios of bioactivity to immunoreactivity of FSH. No significant differences were found in the isoelectric-focusing profiles or bioactivity to immunoreactivity ratios of pituitary FSH in animals treated with testosterone alone or in combination with antagonist. The results demonstrate that testosterone not only maintained the synthesis of both bioactive and immunoreactive FSH in male rats, but also influences the molecular composition of pituitary FSH. These effects of testosterone on pituitary FSH appear not to be mediated through hypothalamic GnRH.

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Phagocytosis by Human Monocytes

Blood ◽

10.1182/blood.v32.3.423.423 ◽

1968 ◽

Vol 32 (3) ◽

pp. 423-435 ◽

Cited By ~ 93

Author(s):

MARTIN J. CLINE ◽

ROBERT I. LEHRER

Keyword(s):

Rna Synthesis ◽

Glucose Utilization ◽

Human Monocytes ◽

Igg Antibody ◽

Enhanced Production ◽

Serum Fractions ◽

Particle Ingestion ◽

Ph Range ◽

Total Glucose

Abstract Human monocytes isolated from peripheral blood phagocytized polystyrene particles, Escherichia coli, Staphylococcus aureus, Candida albicans, Cryptococcus neoformans, and red cells coated with IgG antibody in vitro. The ingestion of fungi and bacteria required serum or serum fractions. Ingestion of antibody-coated erythrocytes was inhibited by whole serum or IgG, but not by IgM or albumin. Adherence to a surface appeared to be critical for erythrophagocytosis but not for ingestion of bacteria. Phagocytosis occurred over a broad pH range and in the presence of inhibitors of oxidative metabolism and of RNA synthesis. Phagocytosis was significantly depressed by inhibitors of glycolysis (iodoacetate and fluoride) and by chloroquine at 1 x 10-5M. Particle ingestion by monocytes was accompanied by enhanced production of C14O2 from glucose-1-C14, but was not associated with a detectable increase in total glucose utilization, incorporation of radioactive uridine into RNA, or incorporation of amino acids into protein.

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Isolation, Partial Purification and Characterisation of A Plasminogen Activator Produced by Endothelial Cells in Vitro

10.1055/s-0039-1687390 ◽

1979 ◽

Author(s):

W.E. Laug

Keyword(s):

Endothelial Cells ◽

Proteolytic Activity ◽

Culture Medium ◽

Partial Purification ◽

Diisopropyl Fluorophosphate ◽

Cell Lysate ◽

Endothelial Cell Cultures ◽

Ph Range ◽

Confluent Endothelial Cell

Cloned endothelial cells obtained from the aorta of 1-2 day old calves produced high fibrinolytic activity, which was 90% dependent upon the presence of plasminogen when grown on 125 I fibrin coated dishes. High plasminogen-dependent proteolytic activity was also demonstrated in the cell lysate and in the culture medium of the cells. The production and secretion of this prtitease were found to increase during the log phase of cell growth and to reach a maximum at con fluency. Thereafter they remained constantly high. This protease, partially purified from the culture medium of confluent endothelial cell cultures, is aiginine specific and activates plasminogen by piOteolytic cleavage to plasmin. Its proteolytic activity which is highest in the pH range of 7.5 to 8.0 is irreversibly inhibited by diisopropyl fluorophosphate, suggesting that it is a serine protease. The molecular weight of this protease is approximately S2000.

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DEVELOPMENT AND EVALUATION OF MUCOADHESIVE NANOGEL OF NEVIRAPINE FOR VAGINAL APPLICATION

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2019v11i3.32353 ◽

2019 ◽

pp. 144-149

Author(s):

Sheikh Sofiur Rahman ◽

ABDUL BAQUEE AHMED

Keyword(s):

Hiv Infection ◽

Ex Vivo ◽

Vaginal Fluid ◽

Content Uniformity ◽

Vaginal Mucosa ◽

Salting Out ◽

Zero Order Kinetics ◽

The Stability ◽

Ph Range

Objectives: The main objective of this study was to develop and evaluate Nevirapine nanoparticle loaded mucoadhesive gel (NVP-Np mucoadhesive gel) for vaginal application for the treatment of HIV infection. Methods: NVP loaded nanoparticles were prepared by salting out method followed by incorporation in different gel bases to produce NVP-Np mucoadhesive gel The prepared gels were evaluated for their physicochemical parameters, rheological characteristics, mucoadhesion, in-vitro drug release and ex-vivo permeation of drug across porcine vaginal mucosa. Results: The result of FT-IR and DSC study confirmed the absence of incompatibility of NVP with excipients used in the formulations. The particle size of the prepared NVP-Np was found to be 243.8 ± 3.15 nm, a polydispersity index (PI) of 0.787± 0.002 and zeta potential value -17.12 mV, which revealed the stability of nanoparticles. All the formulations showed good homogeneity, spreadability, physical appearance and content uniformity. The pH of the mucoadhesive gel formulations was in the range of 3.70 ± 0.03 to 4.56 ± 0.02, which lies in the normal pH range of the vaginal fluid. The cumulative amounts permeated at 6 h were 832.23 ± 63.45 μg/cm2 , 592.13 ± 82.55 μg/cm2 and 941.32 ± 81.10 μg/cm2 from F1(1% Chitosan), F2(1% Carbopol 974P) and F3 (1% HPMC K100M ) respectively. A linear relationship [r2 > 0.9 (0.97 n 0.99)] was observed between the percentage cumulative amount permeated and time, indicating zero order kinetics. Conclusion: In conclusion, NVP-Np mucoadhesive gel was prepared successfully using salting out followed by a homogenization technique for vaginal application of NVP for the prophylaxis of HIV infection.

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