Altered Na+/H+ exchanger isoform 1 activity in immortalized lymphoblasts from women with pre-eclampsia: evidence for an intermediate phenotype

2002 ◽  
Vol 103 (5) ◽  
pp. 503-509 ◽  
Author(s):  
V.M. LEE ◽  
P.A. QUINN ◽  
S.C. JENNINGS ◽  
A.W.F. HALLIGAN ◽  
L.L. NG
Keyword(s):  
Intracellular Ph ◽  
Blood Cells ◽  
Post Partum ◽  
White Blood Cells ◽  
Intermediate Phenotype ◽  
Protein Abundance ◽  
Acetoxymethyl Ester ◽  
Abnormal Volume ◽  
In Cells

Previous studies have demonstrated a raised Na+ content in leucocytes isolated from women with pre-eclampsia. Increased Na+/H+ exchanger activity is one membrane transport abnormality that may contribute to this phenomenon and may be implicated in the abnormal volume homoeostasis and hypertension associated with the disease. Increased Na+/H+ exchanger activity has been documented in nucleated white blood cells from both pre-eclamptic and post-partum pre-eclamptic women, and may suggest the importance of genetic influences on exchanger activity. In the present study, we used lymphoblasts from women with pre-eclampsia and from age- and gestation-matched normotensive pregnant controls to determine Na+/H+ exchanger activity and intracellular resting pH using fluorimetry and the pH-sensitive dye BCECF-AM [bis(carboxyethyl)carboxyfluorescein acetoxymethyl ester]. Determination of Na+/H+ exchanger protein abundance was performed by Western blotting. Intracellular pH was not significantly different in cells from pre-eclamptic women compared with those from normotensive controls. Na+/H+ exchanger activity was measured when the intracellular pH was clamped at 6.0, and was found to be significantly higher in cells from pre-eclamptic women (20.77±0.92mmol·min-1·l-1) compared with those from normotensive controls (15.22±0.92mmol·min-1·l-1; P = 0.001). Na+/H+ exchanger protein abundance was established to be similar in the two subject groups, suggesting that the turnover number for the Na+/H+ exchanger is increased in the women with pre-eclampsia. These changes in Na+/H+ exchanger activity indicate the importance of genetic factors in determining this particular phenotype, since in this cell culture model of pre-eclampsia it is likely that environmental or hormonal influences present in vivo would have declined. Overactivity of the Na+/H+ exchanger may contribute to the raised intracellular Na+ concentration reported previously in white blood cells from women with pre-eclampsia.

scholarly journals In-vivo Antiplasmodial Effect of Dihydroartemisinin-Piperaquine-Nitrofurantoin on Plasmodium berghei- Infected Mice

2021 ◽  
pp. 12-20
Author(s):  
Udeme O. Georgewill ◽  
Festus Azibanigha Joseph ◽  
Elias Adikwu
Keyword(s):  
Plasmodium Berghei ◽  
Blood Cells ◽  
Hematological Parameters ◽  
White Blood Cells ◽  
Positive Control ◽  
Negative Control ◽  
Antiplasmodial Activity ◽  
Significant Difference ◽  
Tract Infections

Nitrofurantoin (NT) used for the treatment of urinary tract infections may have antiplasmodial activity. Dihydroartemisinin-piperaquine (DP) is an artemisinin based combination therapy used for the treatment of malaria. This study evaluated the antiplasmodial effect of dihydroartemisinin-piperaquine-nitrofurantoin (DP-NT) on mice infected with Plasmodium berghei. Adult Swiss albino mice (30-35 g) of both sexes were used. The mice were randomly grouped, inoculated with Plasmodium berghei, and treated orally with DP (1.7/13.7 mg/kg), NT (57.1 mg/kg) and DP-NT (1.71/13.7/ 57.1 mg/kg), respectively using curative, prophylactic and suppressive tests. The negative control was orally treated with normal saline (0.3 mL), while the positive control was orally treated with chloroquine CQ (10mg/kg). After treatment, blood samples were collected and evaluated for percentage parasitemia, inhibitions and hematological parameters. Liver samples were evaluated for histological changes. The mice were observed for mean survival time (MST). Treatment with DP-NT decreased parasitemia levels when compared to individual doses of DP and NT with significant difference observed at p<0.05. DP-NT prolonged MST when compared to individual doses of DP and NT with significant difference observed at p<0.05. The decrease in packed cell volume, red blood cells, hemoglobin and increase in white blood cells in parasitized mice were significantly restored by DP-NT  when compared to individual doses of DP and NT with difference observed at p<0.05. DP-NT eradicated liver Plasmodium parasite.  NT remarkably increased the antiplasmodial activity of DP. DP-NT may be used for the treatment of malaria.

Measurement of intracellular pH in fungal hyphae using BCECF and digital imaging microscopy: Evidence for a primary proton pump in the plasmalemma of a marine fungus

1990 ◽  
Vol 96 (4) ◽  
pp. 731-736
Author(s):  
JULIA M. DAVIES ◽  
C. BROWNLEE ◽  
D. H. JENNINGS
Keyword(s):  
Intracellular Ph ◽  
Proton Pump ◽  
Marine Organism ◽  
Marine Fungus ◽  
Primary Proton ◽  
Acetoxymethyl Ester ◽  
Fluorescence Ratio Imaging ◽  
Negative Membrane Potential ◽  
Ratio Imaging

The facultative marine fungus, Dendryphiella salina, has the most negative membrane potential yet recorded for a marine organism. The ionic basis for this is thought to be through the action of a primary proton pump, though there exists the possibility of electrogenic pumping of Na+ or Cl−, given the high ambient concentration of these ions. Fluorescence ratio imaging microscopy with the pH-sensitive fluorescent probe 2′,7′-bis-(2-carboxyethyl)-5(and-6) carboxyfluorescein (BCECF) has been used to estimate intracellular pH. Hyphae loaded readily with BCECF after incubation with the acetoxymethyl ester (BCECF/AM). Mean resting intracellular pH (pH1) was 7.3, calculated by comparing 490/450 nm fluorescence ratios with in vivo calibration curves obtained by pH equilibration using nigericin. Distinct pH compartments could be observed, corresponding to cytoplasmic and smaller vacuolar compartments. Sodium azide reversibly reduced pH1 by an average of 0.51 of a pH unit, though the response varied between individual hyphae. Inhibiting the plasmalemma ATPase with orthovanadate also reversibly decreased pH|. The results support the presence of a proton pump in the plasmamembrane. The energetic and evolutionary implications are discussed.

scholarly journals Gene expression of the heat stress response in bovine peripheral white blood cells and milk somatic cells in vivo

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
J. B. Garner ◽  
A. J. Chamberlain ◽  
C. Vander Jagt ◽  
T. T. T. Nguyen ◽  
B. A. Mason ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heat Stress ◽  
Blood Cells ◽  
White Blood Cells ◽  
Somatic Cells ◽  
Holstein Friesian ◽  
Milk Somatic Cells ◽  
Peripheral White Blood Cells ◽  
Friesian Cows

Abstract Heat stress in dairy cattle leads to reduction in feed intake and milk production as well as the induction of many physiological stress responses. The genes implicated in the response to heat stress in vivo are not well characterised. With the aim of identifying such genes, an experiment was conducted to perform differential gene expression in peripheral white blood cells and milk somatic cells in vivo in 6 Holstein Friesian cows in thermoneutral conditions and in 6 Holstein Friesian cows exposed to a short-term moderate heat challenge. RNA sequences from peripheral white blood cells and milk somatic cells were used to quantify full transcriptome gene expression. Genes commonly differentially expressed (DE) in both the peripheral white blood cells and in milk somatic cells were associated with the cellular stress response, apoptosis, oxidative stress and glucose metabolism. Genes DE in peripheral white blood cells of cows exposed to the heat challenge compared to the thermoneutral control were related to inflammation, lipid metabolism, carbohydrate metabolism and the cardiovascular system. Genes DE in milk somatic cells compared to the thermoneutral control were involved in the response to stress, thermoregulation and vasodilation. These findings provide new insights into the cellular adaptations induced during the response to short term moderate heat stress in dairy cattle and identify potential candidate genes (BDKRB1 and SNORA19) for future research.

scholarly journals HAEMATOLOGICAL AND BLOOD BIOCHEMICAL PARAMETERS OF PRE - AND POST LAMBING PERIODS FOR IRAQI NUAEMIE EWES

2021 ◽  
Vol 52 (4) ◽  
pp. 941-948
Author(s):  
Awad & et al.
Keyword(s):  
Blood Cells ◽  
Biochemical Parameters ◽  
Post Partum ◽  
Hematological Parameters ◽  
White Blood Cells ◽  
Physiological Status ◽  
Blood Samples ◽  
Blood Biochemical ◽  
Post Partum Period ◽  
Biochemical Assessment

The present study designed  to investigate the hematological and blood biochemical changes in pre and post lambing periods in Iraqi Nuaemie ewes. Ten Nuaemie ewes weighed 35-45 kg and aged between 2-3 years were reared in animal's house of Veterinary College / Tikrit University from October-2018 to March-2019, Ten ml of blood samples were collected from each animal during the periods of last gestation month, at lambing and 2 weeks thereafter, Two and half ml of blood samples were collected in EDTA- containing tubes to determine the hematological parameters and the remaining was used to separate serum and stored at -20 °c for blood biochemical assessment. The results   revealed decreased in total red blood cells, haemoglobin and packed cell volume during post lambing period. The total white blood cells count and neutrophils were decreased during the post-partum period, while the lymphocyte was decreased at the day of lambing (50±5.8%). The biochemical parameters exhibited lesser total protein concentrations at the day of lambing (6.5± 1.85 g/dl ) while greater glucose, cholesterol and triglyceride concentrations during post-partum period. The concentration of urea and creatinine increased during the pre-partum period whereas, LDL and HDL concentrations increased in post- lambing period. The minerals concentrations revealed lesser concentrations of Zink and iron during the post-partum period while, copper concentration was greater during similar period. In conclusion, the physiological status of animals have clearly effects on the haematological and biochemical parameters in Iraqi Nuaemie ewes.  

scholarly journals DNA Damaging Effects, Oxidative Stress Responses and Cholinesterase Activity in Blood and Brain of Wistar Rats Exposed to Δ9-Tetrahydrocannabinol

Molecules ◽  
2019 ◽  
Vol 24 (8) ◽  
pp. 1560 ◽  
Author(s):  
Nevenka Kopjar ◽  
Nino Fuchs ◽  
Suzana Žunec ◽  
Anja Mikolić ◽  
Vedran Micek ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Comet Assay ◽  
Stress Responses ◽  
Blood Cells ◽  
Genome Stability ◽  
White Blood Cells ◽  
Brain Cells ◽  
Alkaline Comet Assay

Currently we are faced with an ever-growing use of Δ9-tetrahydrocannabinol (THC) preparations, often used as supportive therapies for various malignancies and neurological disorders. As some of illegally distributed forms of such preparations, like cannabis oils and butane hash oil, might contain over 80% of THC, their consumers can become intoxicated or experience various detrimental effects. This fact motivated us for the assessments of THC toxicity in vivo on a Wistar rat model, at a daily oral dose of 7 mg/kg which is comparable to those found in illicit preparations. The main objective of the present study was to establish the magnitude and dynamics of DNA breakage associated with THC exposure in white blood and brain cells of treated rats using the alkaline comet assay. The extent of oxidative stress after acute 24 h exposure to THC was also determined as well as changes in activities of plasma and brain cholinesterases (ChE) in THC-treated and control rats. The DNA of brain cells was more prone to breakage after THC treatment compared to DNA in white blood cells. Even though DNA damage quantified by the alkaline comet assay is subject to repair, its elevated level detected in the brain cells of THC-treated rats was reason for concern. Since neurons do not proliferate, increased levels of DNA damage present threats to these cells in terms of both viability and genome stability, while inefficient DNA repair might lead to their progressive loss. The present study contributes to existing knowledge with evidence that acute exposure to a high THC dose led to low-level DNA damage in white blood cells and brain cells of rats and induced oxidative stress in brain, but did not disturb ChE activities.

Stimulation of K-C1 cotransport in rat red cells by a hemolytic anemia-producing metabolite of dapsone

1989 ◽  
Vol 256 (2) ◽  
pp. C265-C272 ◽  
Author(s):  
M. Haas ◽  
J. H. Harrison
Keyword(s):  
Red Blood Cells ◽  
Hemolytic Anemia ◽  
Blood Cells ◽  
Pneumocystis Carinii ◽  
Fold Increase ◽  
Sh Groups ◽  
Splenic Uptake ◽  
Stimulation Of ◽  
In Cells

Dapsone, a sulfone compound used in the treatment of leprosy and, more recently, Pneumocystis carinii pneumonia, produces as a major side effect a hemolytic anemia. This anemia is characterized by oxidation of hemoglobin to methemoglobin and increased splenic uptake of red blood cells. Using a rat model, Grossman and Jollow (J. Pharmacol. Exp. Ther. 244: 118-125, 1988) found that dapsone hydroxylamine (DDS-NOH), a dapsone metabolite, is responsible for its hemolytic effect in vivo. DDS-NOH also promotes hemoglobin binding to SH groups on rat red cell membrane proteins (Budinsky et al., FASEB J. 2: A801, 1988). Since the binding of hemoglobin and other reagents (e.g., N-ethylmaleimide) to membrane SH groups has been associated with increased K transport in red blood cells, we examined the effect of DDS-NOH on K efflux from rat red blood cells in vitro. Cells shrink when exposed to DDS-NOH (100 microM) in media with plasma-like ionic composition. This shrinkage is prevented if extracellular K is raised to 110 mM or if intra- and extracellular Cl are replaced by methylsulfate (MeSO4), suggesting involvement of a K-Cl cotransport pathway. Indeed, 100 microM DDS-NOH produces a 4- to 5-fold increase in K efflux in cells containing Cl but less than a 2-fold increase in cells containing MeSO4. This stimulatory effect is specific for K; Na efflux is slightly inhibited by 100 microM DDS-NOH. The concentrations of DDS-NOH required for half-maximal stimulation of Cl-dependent K efflux (53 microM) is similar to its half-maximal hemolytic concentration in rats (approximately 100 microM). Furthermore, the stimulation of Cl-dependent K efflux by DDS-NOH is greater than 80% reversed by subsequent treatment of the cells with dithiothreitol, suggesting involvement of SH groups. Our results indicate that DDS-NOH exposure stimulates an apparent K-Cl cotransport in rat red blood cells, resulting in cell shrinkage under physiological ionic conditions. Since shrinkage of red blood cells renders them less deformable (Mohandas et al., J. Clin. Invest. 66: 563-573, 1980), this suggests a pathophysiological mechanism whereby DDS-NOH exposure in vivo could promote increased splenic uptake of red blood cells and hemolytic anemia.

Genotoxic effects in human white blood cells following styrene (in vivo) and styrene oxide (in vitro) exposure

1998 ◽  
Vol 95 ◽  
pp. 41
Author(s):  
B. Marczynski ◽  
M. Peel ◽  
P. Rozynek ◽  
J. Elliehausen ◽  
M. Korn ◽  
...  
Keyword(s):  
Blood Cells ◽  
Styrene Oxide ◽  
White Blood Cells ◽  
Genotoxic Effects ◽  
In Vitro Exposure ◽  
Human White Blood Cells
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