scholarly journals The pattern of osmotic regulation in larvae of the mosquito Culiseta inornata

1984 ◽  
Vol 113 (1) ◽  
pp. 133-141 ◽  
Author(s):  
M. Garrett ◽  
T. J. Bradley
Keyword(s):  
Sea Water ◽  
Saline Water ◽  
Mosquito Species ◽  
Full Range ◽  
Chloride Ion ◽  
Malpighian Tubules ◽  
Osmotic Concentration ◽  
Culiseta Inornata

Larvae of Culiseta inornata (Williston) can survive and complete development in dilutions of sea water ranging from 50–700 mosmol kg-1. The larvae hyperregulate with regard to haemolymph osmotic concentration in dilute media (50–400 mosmol kg-1) and osmoconform when external salinities exceed 400 mosmol kg-1. This pattern of osmoregulation is distinct from that observed in freshwater and saline-water mosquito species. We propose that mosquitoes exhibiting this osmoregulatory pattern should be described as ‘brackish-water’ species. Larvae of Culiseta inornata are able closely to regulate both sodium and chloride ion concentrations in the haemolymph over the full range of salinities tested (50–750 mosmol kg-1). The Malpighian tubules produce an isosmotic, potassium-rich fluid. In vitro and in vivo sampling of rectal fluids demonstrates that rectal secretions are isosmotic or only slightly hyperosmotic to the haemolymph and the surrounding saline media, and that they are isotonic with regard to sodium.

scholarly journals The Effectiveness of Candidate Probiotic Bacteria to Control Vibriosis Disease

2017 ◽  
Vol 5 (2) ◽  
pp. 1
Author(s):  
Mulyati Mulyati ◽  
Suryati Suryati ◽  
Irfani Baga
Keyword(s):  
Survival Rate ◽  
Sea Water ◽  
Challenge Test ◽  
Probiotic Bacteria ◽  
Pathogenicity Test ◽  
Inhibitory Ability ◽  
Significant Difference ◽  
Portunid Crab

The study aims to isolate, characterize, and examine probiotic bacteria's inhibitory ability against Vibrio harveyi bacteria, both in-vitro and in vivo. Methods used in the study consist of 1) An Isolation of Candidate Probiotic Bacteria, 2) An Antagonistic Test of Candidate Probiotic Bacteria in vitro, 3) An Identification of Bacteria, 4) A Pathogenicity Test of Candidate Probiotic Bacteria, 5) An Antagonistic Test of Candidate Probiotic Bacteria against V. harveyi in vivo. According to the isolation of candidate probiotic bacteria, there are 18 isolated candidate probiotic. After being tested for its inhibitory ability in vitro, there are 8 isolates with zone of inhibition as follows: isolate MM 7 from intestine (22 mm), isolate MM 6 from intestine (12 mm), isolate MM 10 from sea water (10 mm), isolate MM 5 from intestine (9 mm), isolate MM 4 from intestine (8 mm), isolate MM 3 from intestine (7 mm), isolate MM 2.2 from intestine (7 mm), isolate MM 2.1 from intestine (7 mm). Eight genera of the candidate probiotic bacteria is derived from Portunid crab, they are Staphylococcus, Streptococcus, bacillus, vibrio, Alcaligenes, Lactobacillus, micrococcus. Before proceeding the V. harveyi bacterial challenge test in vivo, three potential isolates consisting of MM6, MM7 and MM10 as the probiotic bacteria are pathogenicity-tested against V. harveyi. The survival rate of Portunid crab on pathogenicity test using MM6, MM7 and MM10 generates 91.11-100%, while the control generates 100% survival rate. Variance analysis result through post-hoc Tukey's Honest Significant Difference (HSD) test at 95% confidence interval indicates that isolate MM7 and MM10 are significantly able to increase hatchling Portunid crab's survival rate.

The anterior and posterior ‘stomach’ regions of larval Aedes aegypti midgut: regional specialization of ion transport and stimulation by 5-hydroxytryptamine

1999 ◽  
Vol 202 (3) ◽  
pp. 247-252 ◽  
Author(s):  
T.M. Clark ◽  
A. Koch ◽  
D.F. Moffett
Keyword(s):  
Steady State ◽  
Aedes Aegypti ◽  
Ion Transport ◽  
Malpighian Tubules ◽  
Transport Mechanisms ◽  
Regional Specialization ◽  
Negative Deflection ◽  
Electrical Polarity

The ‘stomach’ region of the larval mosquito midgut is divided into histologically distinct anterior and posterior regions. Anterior stomach perfused symmetrically with saline in vitro had an initial transepithelial potential (TEP) of −66 mV (lumen negative) that decayed within 10–15 min to a steady-state TEP near −10 mV that was maintained for at least 1 h. Lumen-positive TEPs were never observed in the anterior stomach. The initial TEP of the perfused posterior stomach was opposite in polarity, but similar in magnitude, to that of the anterior stomach, measuring +75 mV (lumen positive). This initial TEP of the posterior stomach decayed rapidly at first, then more slowly, eventually reversing the electrical polarity of the epithelium as lumen-negative TEPs were recorded in all preparations within 70 min. Nanomolar concentrations of the biogenic amine 5-hydroxytryptamine (5-HT, serotonin) stimulated both regions, causing a negative deflection of the TEP of the anterior stomach and a positive deflection of the TEP of the posterior stomach. Phorbol 12,13-diacetate also caused a negative deflection of the TEP of the anterior stomach, but had no effect on the TEP of the posterior stomach. These data demonstrate that 5-HT stimulates region-specific ion-transport mechanisms in the stomach of Aedes aegypti and suggest that 5-HT coordinates the actions of the Malpighian tubules and midgut in the maintenance of an appropriate hemolymph composition in vivo.

Metabolic importance of Na+/K+-ATPase activity during sea urchin development.

1997 ◽  
Vol 200 (22) ◽  
pp. 2881-2892 ◽  
Author(s):  
P Leong ◽  
D Manahan
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Sea Urchin ◽  
Sodium Pump ◽  
Sea Water ◽  
Strongylocentrotus Purpuratus ◽  
Lytechinus Pictus ◽  
Stimulation Of

Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+-ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+-ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development.

The effect of external salinity on drinking rate and rectal secretion in the larvae of the saline-water mosquito Aedes taeniorhynchus

1977 ◽  
Vol 66 (1) ◽  
pp. 97-110
Author(s):  
T. J. Bradley ◽  
J. E. Phillips
Keyword(s):  
Sea Water ◽  
External Medium ◽  
Narrow Range ◽  
Saline Water ◽  
Fluid Secretion ◽  
Osmotic Concentration ◽  
Drinking Rate ◽  
Aedes Taeniorhynchus ◽  
External Salinity ◽  
Kinetics Of

1. The drinking rate of the saline-water mosquito larva Aedes taeniorhyncus (100 nl.mg-1.h-1) is unaffected by the salinity of the external medium, but is directly proportional to the surface area of the animal. 2. Haemolymph Na+, Mg2+, K+, Cl-, SO42- and osmotic concentrations were measured in larvae adapted to 10%, 100% and 200% seawater and were found to be regulated within a narrow range. 3. With the exception of potassium, ionic concentrations in rectal secretion were found to increase with increasing concentrations of the sea water in which larvae were reared. 4. The osmotic concentration of rectal secretion was unaffected by changes in haemolymph osmotic concentration but did rise when sodium or chloride concentrations of the haemolymph were increased. High levels of these ions also stimulated the rate of fluid secretion. 5. Transport of chloride and sodium by the rectum exhibits the kinetics of allosteric rather than classical enzymes.

Regulation of rectal secretion in saline-water mosquito larvae living in waters of diverse ionic composition

1977 ◽  
Vol 66 (1) ◽  
pp. 83-96 ◽  
Author(s):  
T. J. Bradley ◽  
J. E. Philips
Keyword(s):  
Chemical Composition ◽  
External Medium ◽  
Saline Water ◽  
Ionic Composition ◽  
Malpighian Tubules ◽  
Fluid Composition ◽  
Mosquito Larvae ◽  
Major Site ◽  
Osmotic Concentration ◽  
Anal Papillae

1. Larvae of the saline-water mosquito Aedes campestris were adapted to three waters, all having an osmotic concentration of 700 mOsm, but differing in ionic rations. The (Na+Mg) SO4 medium was much moretoxic than the NAHCO3 or the NaCl media. 2. Ionic and osmotic concentrations of haemolymph and rectal secretion were measured in larvae adapted to all three media. The ratio of ionic concentrations in the rectal secretion reflected those in the external medium to which the larvae had been adapted, with the exception of SO42-, which was possibly replaced by HCO3-in the secretion. These differences in rectal fluid composition persisted even though all ligated recta were bathed in the same artificial haemolymph. 3. The Malpighian tubules were found to be the major site of SO42- excretion. In media containing high levels of NA+, Mg2+, K+, Cl- and HCO3-, the rectum secreted a hyperosmotic fluid containing these ions at concentrations several times greater than those found in the haemolymph. 4. These data provide the basis for speculation on the functioning of anal papillae in waters of diverse chemical composition.

scholarly journals Extracellular accumulation of proline, serine and trehalose in the haemolymph of osmoconforming brackish-water mosquitoes

1987 ◽  
Vol 129 (1) ◽  
pp. 231-238 ◽  
Author(s):  
M. A. Garrett ◽  
T. J. Bradley
Keyword(s):  
Organic Compounds ◽  
Brackish Water ◽  
Sea Water ◽  
Full Range ◽  
Culex Tarsalis ◽  
Osmotic Concentration

Larvae of Culex tarsalis, a mosquito, are capable of surviving and developing in dilutions of sea water ranging from 0 mosmol l-1 to 700 mosmol l-1. In waters more dilute than 400 mosmol l-1, the larvae osmoregulate, whereas in those more concentrated than 400 mosmol l-1, the osmotic strength of the haemolymph parallels that of the medium, i.e. the larvae osmoconform. Over the full range of external concentrations tested, the larvae regulate the levels of Na+, K+, Mg2+, Ca2+ and Cl- in the haemolymph. Analyses of haemolymph samples from larvae adapted to media of 50 mosmol l-1 or 600 mosmol l-1 indicate that the increase in haemolymph osmotic concentration observed in media above 400 mosmol l-1 is due to the accumulation of organic compounds, particularly proline, serine and trehalose.

Long-term Adaptations of Sabella Giant Axons to Hyposmotic Stress

1978 ◽  
Vol 75 (1) ◽  
pp. 253-263
Author(s):  
J. E. TREHERNE ◽  
Y. PICHON
Keyword(s):  
Sea Water ◽  
Potassium Concentration ◽  
Resting Potential ◽  
Sodium Permeability ◽  
Bathing Medium ◽  
Appreciable Change ◽  
Axon Membrane ◽  
Giant Axons

Reprint requests should be addressed to Dr Treherne. Sabella is a euryhaline osmoconformer which is killed by direct transfer to 50% sea water, but can adapt to this salinity with progressive dilution of the sea water. The giant axons were adapted to progressive dilution of the bathing medium (both in vivo and in vitro) and were able to function at hyposmotic dilutions (down to 50%) sufficient to induce conduction block in unadapted axons. Hyposmotic adaptation of the giant axon involves a decrease in intracellular potassium concentration which tends to maintain a relatively constant resting potential during adaptation despite the reduction in external potassium concentration. There is no appreciable change in the intracellular sodium concentration, but the relative sodium permeability of the active membrane increases during hyposmotic adaptation. This increase partially compensates for the reduction in sodium gradient across the axon membrane, during dilution of the bathing media, by increasing the overshoot of the action potentials recorded in hyposmotically adapted axons.

scholarly journals Anti-Inflammatory Properties of Mineral-Balanced Deep Sea Water in In-Vitro and In-Vivo Models of Inflamed Intestinal Epithelium

2020 ◽  
Vol 10 (15) ◽  
pp. 5183
Author(s):  
Jain Nam ◽  
Kyeong Jin Kim ◽  
Geonhee Park ◽  
Byeong Goo Kim ◽  
Gwi-Hwa Jeong ◽  
...  
Keyword(s):  
Deep Sea ◽  
Sea Water ◽  
Cell Model ◽  
Mineral Waters ◽  
In Vivo Models ◽  
Deep Sea Water ◽  
High Calcium ◽  
A Cell

This study aimed to determine the effect of deep-sea water (DSW)-derived mineral waters on intestinal health, using a cell model and a dextran sulfate sodium (DSS)-induced enteritis mouse model. DSW was desalted and minerals were added to generate mineral waters that were classified as trace mineral (TM), high magnesium (HM), high magnesium low salt (HMLS), and high magnesium high calcium (HMHC), using a tabletop electrodialysis device. Caco-2 cells cocultured with Raw264.7 cells were either pre-treated or not with the four water groups, and inflammation was induced by treatment with lipopolysaccharide (LPS). Compared to LPS-treated Caco-2 cells, HMLS-cotreated cells maintained high transepithelial electrical resistance, similar to control cells. FITC-dextran permeability was lower in HMLS-treated than in other cells. In vivo, in comparison to DSS-treated mice, colon shortening was inhibited, and disease activity and colon injury were suppressed in HMLS-cotreated mice. RNA-seq of colonic tissues revealed that inflammatory gene expression was similar among the control and HMLS mice, and DSS-induced expression of inflammation-related genes such as TNF-α and NOS2 and inflammatory chemokine genes was suppressed. Our findings suggest that DSW-derived mineral water intake can help reduce colitis symptoms, and the effects may be partially regulated by magnesium and other minerals.

New targeted anti CDK4/6 peptide MM-D37K.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e13545-e13545 ◽  
Author(s):  
Vladimir Konstantinovich Bozhenko ◽  
Tatyana Michailovna Kulinich ◽  
Elena Aleksandrovna Kudinova ◽  
Andrey Boldyrev ◽  
Vladimir Alekseevich Solodkij
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Full Range ◽  
Mrna Level ◽  
Xenograft Model ◽  
Specific Inhibitor ◽  
Antitumor Effects ◽  
Mcf 7

e13545 Background: MM-D37K is a synthetic peptide which consists of p16INK4a-specific inhibitor of complex cyclin D- CDK4 and CDK6 and cell penetrating peptide (CPP) – Antp (Penetratin). We investigated in vitro and in vivo cytotoxic, cytostatic and antitumor activity of MM-D37K. The level of cyclin A, Ki67,bax, bcl-2 and pRb phosphorylation was investigated. Full range of Toxicology tests and Pharmacokinetics experiments in mice, rats and rabbits were performed. Methods: Different cell lines (Jurcat, Raji, A549, MCF-7, Hct-116, Ht-29, HEK293) were incubated with 0.1-100 mM MM-D37K for 24-48 hrs. Proliferation (MTT), DNA-content, cell cycle (flow cytometry) and mRNA level of appropriate proteins (RT PCR) were investigated. In vivo experiments were conducted on xenograft model of HCT116, A-549 at concentration 5 and 10 mg/kg of MM-D37K. Toxicology experiments were made under RF Law and included 3 types of animals. LC-MS MMD37K method of detection in plasma was developed. Results: MM-D37K prevented pRb phosphorilation and proliferation activation in all investigated cell lines. Cell cycle was blocked in G1 phase. Cytostatic effect did not depend on p16 mutation or expression. MM-D37K induced apoptosis in 20-82% of investigated cells at 40 mM with lowest level for MCF-7. LD10 for rats was 100 mg/kg and no deaths were registered for rabbits (highest dose was 50 mg/kg). Concentration of MMD-37K in plasma after 2 min and bolus i.v. injection in dose 10 mg/kg was 72.16±5.64 mcg/ml. Concentration decreased in two phases. 1st – t1/2 = 2.39±0.39 min and for 2nd t1/2=2.39±0.39 hr. Antitumor effects in xenograft model were 53% for A-549 and 67% for HCT116. Conclusions: Our results proved cytotoxic, cytostatic and antitumor effects of MM-D37K in investigated cell lines in vitro and in vivo. Toxicological and pharmacokinetics results allow us recommend for I/IIa Phase clinical trial. (Support: MetaMax Ltd., RFFI, Minpromtorg RF.)

scholarly journals Evaluation of a mathematical model of rumen digestion and an in vitro simulation of rumen proteolysis to estimate the rumen-undegraded nitrogen content of feedstuffs

1983 ◽  
Vol 50 (3) ◽  
pp. 555-568 ◽  
Author(s):  
U. Krishnamoorthy ◽  
C. J. Sniffen ◽  
M. D. Stern ◽  
P. J. Van Soest
Keyword(s):  
Mathematical Model ◽  
Rumen Fluid ◽  
Full Range ◽  
Enzyme Concentration ◽  
Total N ◽  
Protease Enzyme ◽  
Residual N

1. Twelve grain mixtures, one lucerne (Medicago sativa) hay and one maize silage which had been used in mixed diets for which dietary nitrogen undegraded in the rumen (UDN) had been estimated with duodenally-cannulated cows, were studied. Total N in the feeds was fractionated into pool A (N soluble in borate–phosphate buffer), pool B (total N–(pool A + pool C)) and pool C (acid-detergent-insoluble N or residual N after 24 h incubation in protease solution).2. N solubilization in protease solution containing 6·6 units/ml (substrate-saturating enzyme concentration) indicated the presence of subfractions in pool B, with different rates of solubilization. Such subfractions were not detectable from in situ, Dacron bag, estimates of N solubilization.3. UDN was estimated using a dynamic mathematical model and rate-constants obtained from N solubilization in protease solution or in situ.For three grain mixtures tested using the protease technique the model predicted UDN values of 7, 10 and 12% compared with values of 47, 66 and 59% estimated in vivo. The full range of experimental feeds was tested using the in situtechnique and UDN values predicted by the model were used to derive UDN values for twelve mixed diets. The latter values were significantly but not closely correlated with those determined in vivo (r2 0·41, P < 0·05).4. An attempt was made to simulate rumen proteolysis in vitro by choosing a protease enzyme concentration (0·066 units/ml) providing a proteolytic activity similar to that of whole rumen fluid. The experimental samples of feed were subjected to simulated rumen proteolysis for 18 or 48 h to resemble the mean retention times in the rumen for grain mixtures and roughages respectively. The residual N at the end of incubation was considered as an estimate of UDN. The UDN values estimated from simulated rumen proteolysis and those determined in vivo for twelve mixed diets were in close agreement (r2 0·61, P < 0·01).5. Simulated rumen proteolysis can serve as a simple, rapid and sensitive method to estimate UDN in a variety of feedstuffs.

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