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Localization of high benzaldehyde dehydrogenase activity in rat upper gastrointestinal tract mucosa: a quantitative histochemical study.

An unusual aldehyde dehydrogenase (AlDH) phenotype, histochemically similar to the "tumor-associated" AlDH appearing during rat hepatocarcinogenesis, was detected in normal rat upper gastrointestinal tract tissues. This phenotype is characterized by high activities with aromatic substrates, i.e., benzaldehyde (Bz) and NADP. Frozen sections of GI tract tissue from normal rats and from liver nodules induced by a Solt-Farber protocol were evaluated for AlDH activity. A sensitive, high-resolution procedure was used in which sections are pre-incubated in nitroblue tetrazolium and then incubated at 20 degrees C in a viscous polyvinyl alcohol medium containing buffer, phenazine methosulfate, sodium azide, substrate, co-enzyme, and nitroblue tetrazolium. Incubation at a suboptimal pH of 7.0 was found to improve retention of the final reaction product and the linearity with time. Activity was quantitated by computer-assisted microscopic photometry. Intense BzDH-NADP activity was localized in the squamous epithelium of the tongue, esophagus, and fore-stomach, and in the glandular pit cells of the glandular stomach; this activity was not evident in the submucosa, muscle walls, and vessels. Little if any BzDH-NADP activity was observed in the small or large intestine, pancreas, and liver. AlDH in upper GI tissues and in liver nodules shared three characteristics: a sharp localization; a preference for Bz and NADP compared to the aliphatic substrate acetaldehyde and NAD; and a high co-enzyme-independent activity in the presence of Bz.

Aldehyde Oxidoreductases↗

Effect of lactation counselling on subclinical mastitis among Bangladeshi women.

Subclinical mastitis, defined as raised milk sodium/potassium (Na/K) in the absence of clinical symptoms, is associated with poor infant weight gain and increased risk of mother-to-child HIV transmission. Similarly to clinical mastitis, subclinical mastitis appears to have multiple causes, one being poor lactation practice. We investigated whether a single session of lactation counselling around the time of delivery could decrease subclinical mastitis among women in rural Bangladesh. The counselling messages included the importance of giving colostrum, exclusive breastfeeding, feeding on demand and how to achieve good positioning and attachment. Women who had delivered prior to our instituting the counselling intervention (mean 1 month) constituted the uncounselled group. At 1 month postpartum, sodium and potassium were measured in spot milk samples from each breast by flame photometry and interleukin-8 (IL8) by ELISA. Geometric mean Na/K ratios and the proportion of milk samples with raised Na/K ratio were significantly (p < 0.05) lower in the counselled group [mean 0.42, 95% confidence interval (CI) 0.39-0.45, n = 116 samples; 10% Na/K ratio > 0.6] than in the non-counselled group [mean 0.49, 95% CI 0.45-0.53, n = 127; 25% Na/K ratio > 0.6]. Milk IL8 concentration did not differ between groups. The results show that among Bangladeshi women a simple lactation counselling intervention could improve lactation practice in ways likely to improve infant health.

Adult↗

Nebulizer calibration using lithium chloride: an accurate, reproducible and user-friendly method.

Conventional gravimetric (weight loss) calibration of jet nebulizers overestimates their aerosol output by up to 80% due to unaccounted evaporative loss. We examined two methods of measuring true aerosol output from jet nebulizers. A new adaptation of a widely available clinical assay for lithium (determined by flame photometry, LiCl method) was compared to an existing electrochemical method based on fluoride detection (NaF method). The agreement between the two methods and the repeatability of each method were examined. Ten Mefar jet nebulizers were studied using a Mefar MK3 inhalation dosimeter. There was no significant difference between the two methods (p=0.76) with mean aerosol output of the 10 nebulizers being 7.40 mg x s(-1) (SD 1.06; range 5.86-9.36 mg x s(-1)) for the NaF method and 7.27 mg x s(-1) (SD 0.82; range 5.52-8.26 mg x s(-1)) for the LiCl method. The LiCl method had a coefficient of repeatability of 13 mg x s(-1) compared with 3.7 mg x s(-1) for the NaF method. The LiCl method accurately measured true aerosol output and was considerably easier to use. It was also more repeatable, and hence more precise, than the NaF method. Because the LiCl method uses an assay that is routinely available from hospital biochemistry laboratories, it is easy to use and, thus, can readily be adopted by busy respiratory function departments.

Aerosols↗

Effects of streptozotocin-induced type 1 diabetes mellitus on total protein concentrations and cation contents in the isolated pancreas, parotid, submandibular, and lacrimal glands of rats.

This study investigated the effect of streptozotocin (STZ)-induced type 1 diabetes mellitus (DM) on total protein concentration and levels of sodium (Na+), potassium (K+), magnesium (Mg2+), zinc (Zn2+), copper (Cu2+), calcium (Ca2+), and iron (Fe2+) in the pancreas, parotid, submandibular, and lacrimal glands of the rat, compared to age-matched control animals. Protein concentrations were measured by the Bradford Assay, whereas levels of Na+, K+, Mg2+, Zn2+, Cu2+, Ca2+, and Fe2+ were measured by flame photometry and atomic absorbance spectrophotometry. The results show marked changes in the characteristics of diabetic and control animals. Diabetic rats and their different glands weighed significantly (P < 0.05) less compared to age-matched controls. Diabetic rats also have significantly elevated blood glucose and significantly reduced plasma insulin, compared to controls. The results also show that the concentrations of proteins and levels of cations were significantly (P < 0.05) reduced in the pancreas, parotid, submandibular and lacrimal glands of diabetic rats, compared to glands from age-matched animals. These differences in the cation contents and protein levels in STZ-induced DM in this study, along with supporting evidences from previous studies, may provide evidence for the development of long-term complications of DM including exocrine gland deficiencies.

Animals↗

The 22-kD peroxisomal integral membrane protein in Zellweger syndrome--presence, abundance, and association with a peroxisomal thiolase precursor protein.

The primary genetic defect of Zellweger syndrome may be related to defective synthesis or impaired import of peroxisomal proteins. We analyzed the presence and measured the abundance of the 22-kD peroxisomal integral membrane protein (PMP) in patients with Zellweger syndrome. We determined the intracellular localization of the 22-kD PMP and compared it with the localization of a peroxisomal 44-kD thiolase precursor protein. The 22-kD PMP was quantified by immunoblot analyses in liver tissue (n = 7 patients). Immunoblot signals were evaluated using transmission photometry. The intracellular localization of the 22-kD PMP and the peroxisomal 44-kD thiolase precursor protein were determined by immunoblot analyses on fibroblast subcellular fractions prepared by Nycodenz (n = 5 patients) or sucrose density gradient centrifugation (n = 2 patients). The 22-kD PMP was present and associated with membrane fractions in all patients. Its abundance varied in patients as compared with normal human liver controls. The 22-kD PMP was located in subcellular membrane fractions having a lower density than normal peroxisomes or mitochondria. Using two different gradient techniques, the 22-kD PMP and the peroxisomal 44-kD thiolase precursor protein were found in the same low-density gradient fractions. These results suggest that in Zellweger syndrome peroxisome-like elements containing both the 22-kD PMP and a 44-kD thiolase precursor protein are formed. Globally defective synthesis or import of peroxisomal proteins is therefore unlikely to be the primary genetic defect in the patients we studied.

Acetyl-CoA C-Acetyltransferase↗

Insulin and aldosterone interaction on NA+ and K+ transport in cultured kidney cells (A6).

The effects of aldosterone (A) and insulin (I) on active Na+ transport [short circuit current (Isc)] and basolateral to apical K+ transport were evaluated in cultured epithelia derived from toad kidney (A6). Aldosterone plus insulin (A + I) stimulated Isc approximately 2 times the sum of the increments induced by A and I separately. Time courses of Isc stimulation by A + I in epithelia pretreated with A demonstrated that the synergism began about 80 min after the addition of insulin. The effect of A and I on basolateral to apical K+ transport was evaluated by apical K+ accumulation (flame photometry) in epithelia incubated for 24 h with K+-free medium initially placed on the apical side and various K+ concentrations on the basolateral side. At 4.5 mM basolateral K+, I had virtually no effect on basolateral to apical K+ transport, while both A and A + I produced significant increases (44% and 116%, respectively). Stimulation of basolateral to apical K+ transport by A + I was found to be concentration dependent. The EC50 for A stimulation (fixed I concentration) of K+ transport and Isc was 1-3 X 10(-8) M, which is quite similar to the Kd for nuclear binding of A previously reported. Our findings are consistant with the hypothesis that insulin, in addition to its own effects on Na+ transport, may cause an amplification of the mechanism of action of aldosterone on Na+ and K+ transport. The synergistic responses produced by A + I may represent an important hormonal interaction in regulation of transepithelial Na+ and K+ transport.

Aldosterone↗

Interferon-gamma reduces actin filaments and inhibits thyroid-stimulating hormone-induced formation of microvilli and pseudopods in mouse monolayer thyrocytes.

To study the role of interferon-gamma (IFN gamma), which is released from activated lymphocytes in the development of lymphocytic thyroiditis, we investigated the effects of IFN gamma on the morphology of mouse thyrocytes, using scanning electron microscopy. Thyrocytes were cultured in monolayers for 5-7 days and incubated with TSH (10 mU/ml) for 1 h before fixation. IFN gamma (200 U/ml) was added to the culture medium for 1 h to 5 days before TSH stimulation. Coculture of thyrocytes with IFN gamma for more than 24 h inhibited TSH-induced morphological changes in the thyrocytes; IFN gamma inhibited the increase in the number of and elongation of microvilli and the appearance of pseudopods. IFN gamma also reduced the number of actin filaments in thyrocytes, as confirmed by immunofluorescence photometry. These results suggest that IFN gamma inhibits the morphological response of thyrocytes to TSH stimulation by the reduction of actin filaments.

Actins↗

Activation of the phosphatidylinositol pathway in the primate corpus luteum by prostaglandin F2 alpha.

The current study was designed to investigate the ability of prostaglandin F2 alpha (PGF2 alpha) to activate a second messenger system (phosphatidylinositol pathway) in corpora lutea (CL) of rhesus monkeys. Activation of this pathway was assessed by monitoring the hydrolysis of phosphatidylinositol to inositol phosphates. Since inositol triphosphate mobilizes intracellular Ca2+, intracellular free calcium concentrations ([Ca2+]i) were also assessed in individual cells by fura-2 fluorescence photometry. These responses to PGF2 alpha were measured in luteal cells collected from nonpregnant rhesus monkeys. CL were collected during the early (days 4-5 after estimated LH surge; n = 4), mid (days 8-9; n = 4), and late (days 13-14; n = 5) luteal phase and 1 day after in vivo hCG treatment (15 IU/dose, morning and evening), which began during the midluteal phase (n = 5). PGF2 alpha significantly increased the accumulation of inositol phosphates in all groups (P less than 0.05), except the midluteal phase (P = 0.07). The luteal sensitivity to PGF2 alpha, judged by phosphatidylinositol hydrolysis, was low in the early to midluteal phase compared to that in the late luteal phase and after in vivo hCG treatment. PGF2 alpha also caused a rapid, yet transient, increase in [Ca2+]i in a large proportion of primate luteal cells. The proportion of luteal cells that responded to PGF2 alpha with an increase in [Ca2+]i was smaller (P less than 0.05) in CL collected during the early luteal phase than in the other groups. Luteal progesterone production was inhibited by PGF2 alpha in CL collected after in vivo hCG. CL treated in vivo with hCG also displayed in vitro the largest increases in phosphatidylinositol hydrolysis and [Ca2+]i in response to PGF2 alpha. Therefore, this study demonstrates that PGF2 alpha is a potent activator of the phosphatidylinositol pathway in the primate CL. This activation is augmented as the luteal phase progresses and is influenced by in vivo hCG treatment. This study also provides evidence that the inhibitory effects of PGF2 alpha on progesterone production are associated with the activation of the phosphatidylinositol pathway.

Animals↗

Growth hormone-releasing hexapeptide elevates intracellular calcium in rat somatotropes by two mechanisms.

The actions of GH-releasing hexapeptide (His-D-Trp-Ala-Trp-D-Phe-Lys-NH2 or GHRP-6) on single rat somatotropes were studied using whole cell patch clamp electrophysiology and indo-1 Ca2+ photometry. GHRP-6 elevated intracellular free Ca2+ ([Ca2+]i) in two phases: a rapid transient phase, followed by a persistent phase. Based on its insensitivity to treatments that block Ca2+ entry [removal of external Ca2+, addition of the dihydropyridine Ca2+ channel blocker nitrendipine (1 microM), and the hyperpolarizing action of zero external Na+ or 100 nM somatostatin], the transient elevation is the result of release of Ca2+ from intracellular stores. The half-maximal concentration for the peak [Ca2+]i rise during Ca2+ release was 49 nM GHRP-6. Prior treatment of cells with caffeine (10 mM) or ryanodine (50 microM) abolished or partially occluded GHRP-6-induced Ca2+ release. Simultaneous measurement of [Ca2+]i and membrane current or potential revealed that the transient release of Ca2+ by GHRP-6 activates a voltage-independent Ca(2+)-activated K+ conductance, which transiently hyperpolarizes the somatotrope. The GHRP-6-induced persistent [Ca2+]i elevation is abolished by removal of external Ca2+ or external Na+ or the addition of 1 microM nitrendipine or 100 nM somatostatin, consistent with Ca2+ entry through voltage-dependent Ca2+ channels. In nondialyzed cells (perforated patch recording), we have identified a long-lasting GHRP-6-induced depolarization which may be responsible for the persistent [Ca2+]i elevation.

Animals↗

Sodium-potassium pump activity contributes to the age-related decline in resting metabolic rate.

We examined the hypothesis that a decline in Na-K pump activity contributes to the lower resting metabolic rate (RMR) in older males, independent of the loss of fat-free mass. Plasma and erythrocyte Na and K concentrations were measured using flame photometry. Changes in these concentrations after incubation with and without ouabain were used to calculate erythrocyte Na-K pump rate and rate constant in 27 younger (28 +/- 7 yr) and 25 older (67 +/- 6 yr) men. Older men showed an 18% lower erythrocyte Na-K pump rate constant (0.32 +/- 0.09 vs. 0.39 +/- 0.10 h-1; P < 0.01), a 12% lower RMR (1.10 +/- 0.14 vs. 1.25 +/- 0.14 kcal/min; P < 0.01) and an 8% lower level of fat-free mass (61.7 +/- 5.8 vs. 67.4 +/- 8.1 kg; P < 0.01) relative to younger men. A lower RMR persisted in older men (1.14 +/- 0.12 kcal/min) compared to younger men (1.21 +/- 0.12 kcal/min; P < 0.05) after control for the effect of fat-free mass. No differences in RMR were found, however, between older (1.17 +/- 0.13 kcal/min) and younger men (1.20 +/- 0.13 kcal/min) after controlling for both fat-free mass and the erythrocyte Na-K pump rate constant. A positive relation was noted between RMR and the erythrocyte Na-K pump rate constant, after removing the effects of fat-free mass (partial r = 0.30; P < 0.05). Our results support the conclusions that: 1) the in vivo activity of the Na-K pump is related to RMR, and 2) the age-related reduction in Na-K pump activity is a partial contributor to the decline in RMR in older men.

Adult↗

Procaine, lidocaine, and hypothermia inhibit calcium paradox in glial cells.

The purpose of this study was to determine the effects of local anesthetics (lidocaine and procaine) and hypothermic perfusion on Na+/Ca2+ exchanger activity during reperfusion-induced calcium paradox in vitro. Hypothetically, this work provides a mechanism for reperfusion paradox injury in vitro that may relate to a variety of ischemic insults in vivo. Intracellular calcium ([Ca2+]i) in individual cells was measured using a fura-2 photometry system during protocols previously shown to dramatically and persistently increase [Ca2+]i in a manner that replicates certain phases of reperfusion injury in vivo. Our results suggest that short (< 5.0 min) exposure to hypocalcia prompts subsequent, potentially lethal, sodium-dependent calcium entry via the reverse phase of the Na+/Ca2+ exchanger. The local anesthetics lidocaine or procaine are able to block [Ca2+]i increases when added at the time of reperfusion. Furthermore, since [Ca2+]i is normally increased when local anesthetics are added by themselves, the results suggest that local anesthetics promote the forward mode of Na+/Ca2+ exchange (i.e., Na-dependent calcium removal) in addition to reverse Na+/Ca2+ exchanger blockade. Low temperature (18 degrees C) perfusion or reperfusion also affects exchanger dynamics and attenuates calcium entry during such protocols. These experiments further implicate the Na+/Ca2+ exchanger in reperfusion-mediated cell injury and provide potential ways to therapeutically modify such phenomena in vivo.

Analysis of Variance↗

The effect of nitroglycerin on microvascular perfusion and oxygenation during gastric tube reconstruction.

Esophagectomy followed by gastric tube reconstruction is the surgical treatment of choice for patients with esophageal cancer. Complications of the cervical anastomosis are associated with impaired microvascular blood flow (MBF) and ischemia in the gastric fundus. The aim of the present study was to differentiate whether the decrease in MBF is a result of arterial insufficiency or of venous congestion. To do this we assessed MBF, microvascular hemoglobin oxygen saturation (muHbSo(2)), and microvascular hemoglobin concentration (muHbcon) simultaneously during different stages of gastric tube reconstruction. In 14 patients, MBF was determined with laser Doppler flowmetry, and muHbSo(2) and muHbcon were determined with reflectance spectro- photometry. After completion of the anastomosis, nitroglycerin was applied at the fundus. Although MBF did not change significantly in the pylorus, MBF decreased progressively during surgery in the fundus from 210 +/- 18 Arbitrary Units at baseline (normal stomach) to 52 +/- 9 Arbitrary Units after completion of reconstruction (mean +/- sem; P < 0.05). There was no change in muHbSo(2) and muHbcon during the reconstruction. After application of nitroglycerin, MBF doubled. We conclude that MBF decreases during gastric tube reconstruction but that muHbSo(2) and muHbcon do not. This decrease might be the result of venous congestion, which can partly be counteracted by application of nitroglycerin.

Adult↗

Sodium activity of insect blood: physiological significance and relevance to the design of physiological saline.

The apparent activity coefficients for sodium (gamma'Na) in the blood of six insect species have been calculated from measurements made with sodium-selective electrodes and a flame photometer. In every case gamma Na was significantly lower than that for this cation in free solution (gammaNa). In Periplaneta americana gammaNa varied considerably, during a period of 90 days, so that a relatively constant sodium activity (aNa) was maintained in the blood in the face of large variations in the total sodium content measured by flame photometry. Despite the relative constancy of aNa (of around 0.088M) appreciable fluctuations were observed in the sodium and potassium content of nervous connective over a period of 140 days. The values of aNa and aK were used to devise a satisfactory cockroach saline for use in experiments with isolated nerve cords.

Animals↗

An electrophysiological analysis of extra-axonal sodium and potassium concentrations in the central nervous system of the cockroach (Periplaneta americana L.).

Simultaneous intracellular and sucrose-gap recordings showed, in contrast to previous findings, that the electrical parameters of giant axons were similar to intact and desheathed connectives bathed with the 'extracellular Ringer' of Yamasaki & Narahashi. This implies that the extra-axonal sodium concentration, in situ, is likely to be lower than had been previously supposed. Axonal responses showed that, despite the high blood concentration of 24-2 mM-K+ measured by flame photometry, the effective concentration in the blood was 10-15 mM-K+ which corresponds to the measurements made with potassium-selective electrodes. The activity of the blood potassium ions caused a marked reduction in the amplitude of the action potentials following surgical desheathing or disruption of the blood-brain barrier with hypertonic urea. It is suggested that a regulatory mechanism exists in the central nervous system which counteracts the effects of the high blood potassium level.

Action Potentials↗

Use of Ion-Channel Modulating Agents to Study Cyanobacterial Na(+) - K(+) Fluxes.

Here we describe an experimental design aimed to investigate changes in total cellular levels of Na(+) and K(+) ions in cultures of freshwater filamentous cyanobacteria. Ion concentrations were measured in whole cells by flame photometry. Cellular Na(+) levels increased exponentially with rising alkalinity, with K(+) levels being maximal for optimal growth pH (~8). At standardized pH conditions, the increase in cellular Na(+), as induced by NaCl at 10 mM, was coupled by the two sodium channel-modulating agents lidocaine hydrochloride at 1 microM and veratridine at 100 microM. Both the channel-blockers amiloride (1 mM) and saxitoxin (1 microM), decreased cell-bound Na(+) and K(+) levels. Results presented demonstrate the robustness of well-defined channel blockers and channel-activators in the study of cyanobacterial Na(+)- K(+) fluxes.

Journal Article↗

[Micromethods for the determination of metabolic characteristics in individual nephron segments].

Since a single nephron is a functional unit of the kidney, individual microdissected segments from the nephron would be ideal tissue samples for investigations on renal pharmacology. Several micromethods have enabled researchers to analyze the biochemical and pharmacological characteristics of these nephron segments. Miniaturized cuvettes containing microliter volumes of samples can be applied for general procedures of photometry like Lowry's protein determination. Fluorometry becomes a more sensitive method when enzymatic cycling systems of NAD/NADH or NADP/NADPH are combined, which have been used for assays of enzyme activities or substrate contents in minute biological samples having tissue proteins less than 1 microgram. The microchemiluminescence procedure has been successfully utilized for cellular ATP content or oxygen radical generation. Radioimmunoassay can be used to determine endogenous components such as cyclic nucleotides, eicosanoids, etc. Continuous gradient polyacrylamide microgels prepared in 5- to 10-microliters capillaries have made it possible to quantify the intranephron distribution of cytochrome P-450, xanthine oxidase and superoxide dismutase. As an example of the modern techniques, microscopic fluorometry using Fura-2AM has been established to identify agonist-induced cytosolic free calcium transients.

Ammonia↗

Characterization of carbonic anhydrase isozyme CA2, which is the CAH2 gene product, in Chlamydomonas reinhardtii.

From high-CO2 (5% CO2) grown unicellular green alga, Chlamydomonas reinhardtii, carbonic anhydrase (CA) was isolated by affinity chromatography and characterized. Isolated CA was identified as an isozyme (CA2) which is the product from the second gene CAH2 by peptide sequencing. The CA2 was inactivated by dithiothreitol. This treatment caused dissociation of CA2 into the large (38 kDa) and small subunits (4243 Da). The molecular mass of the CA2 holoenzyme measured by low-angle laser light-scattering photometry and precision differential refractometry combined with gel-filtration HPLC was 87.9 kDa. These results and gene structure indicate that CA2 is a heterotetramer consisting of two large and two small subunits linked by disulfide bonds like CA1, which is the CAH1 gene product. The specific activity of CA2 purified by anion-exchange HPLC was 3300 units per mg protein, which was approximately 1.6 times higher than that of CA1. Therefore, it was concluded that two structurally related isozymes, CA1 and CA2, are present in the wild type cells of C. reinhardtii and differentially regulated by the atmospheric CO2 concentration.

Amino Acid Sequence↗

Effect of erythropoietin treatment on blood pressure and intracellular cation concentrations in maintenance hemodialysis patients.

To assess the effect of recombinant human erythropoietin (EPO) on the factors regulating blood pressure (BP), we determined the hemoglobin level (Hgb), blood viscosity (BV), plasma renin activity (PRA), plasma concentrations of aldosterone (PAC), adrenaline (Ad), noradrenaline (NAd), and atrial natriuretic peptide (ANP), and serum and intracellular concentrations of cations before and after 3 months of EPO treatment (40 units/kg/week of EPO intravenously after each hemodialysis session) in 11 patients undergoing maintenance hemodialysis. Intracellular sodium concentration ([Na+]i) was measured using erythrocytes with flame photometry. EPO treatment was associated with significant increases in Hgb (7.1 +/- 1.4 to 8.4 +/- 1.8 g/dl, p<0.01), mean BP (103 +/- 11.4 to 116 +/- 19.9 mmHg, p<0.01), [Na+]i (4.99 +/- 0.78 to 6.22 +/- 0.96 mmol/l, p<0.01) and BV (1.39 +/- 0.14 to 1.53 +/- 0.18 c.p., p<0.05), but no significant alteration in PRA, PAC, Ad, NAd, ANP, or in the serum concentration of Na+, K+, and Ca2+. The changes in mean BP (deltaMBP) were significantly correlated with delta[Na+]i (R=0.676, p=0.022) and deltaBV (R=0.668, p=0.034), but not with deltaHgb. By multiple regression analysis, delta[Na+]q and deltaBV independently contributed to deltaMBP; deltaMBP=2.27 X delta[Na+]i+32.2 X deltaBV +3.37 (R=0.695). These data suggest that intracellular sodium accumulation as well as increased blood viscosity may be independently involved in the blood pressure elevation after EPO treatment in patients under maintenance hemodialysis. We found no evidence supporting a role of circulating hormonal factors, such as the renin-angiotensin system, adrenaline, or ANP, in the change in blood pressure.

Aged↗