Hypotonic Research Articles

Hospital-acquired hyponatremia: Does the type of fluid therapy affect children admitted to intensive care?

The objective of this study was to evaluate the association between the type of initial fluid therapy used (isotonic or hypotonic solutions) and the development of hyponatremia, the plas ma chlorine values and the tolerance of venous access. Retrospective cohort study in a Pediatric Intensive Care Unit (PICU) of a high complexity hospital. There were included children younger than 15 years old hospitalized during the first semester of 2010 and 2013 who recei ved intravenous maintenance fluid therapy, excluding patients undergoing cardiac surgery, kidney transplant and admissions that lasted less than 24 hours. Epidemiological, comorbidity and admis sion-related data were collected, including type of solution received, sodium and chlorine values in the first 72 hours of hospitalization and the incidence of extravasation of peripheral intravenous lines. 111 children were included; 68 children (61.3%) were treated with hypotonic solutions and 43 (38.7%) with isotonic solutions. There were no differences in pathology and severity, and also in the volume of fluid received. Among the patients who received hypotonic solutions, 28 (41.2%) de veloped hyponatremia, wich was moderate (Na <130 mEq/Kg) in 11 cases, compared with 8 children (18.6%) who received isotonic solutions, with only one case of moderate hyponatremia (p = 0.027). No cases of hypernatremia were recorded, and there were no differences in plasma chlorine values. There was also no increased frequency of venous access loss in patients treated with isotonic solutions (4.7% versus 7.4%, p = 0.704). Treatment with initial isotonic solutions in children hos pitalized in PICU is associated with a lower incidence and severity of hyponatremia, without changes in the plasma chlorine values and it is well tolerated by peripheral intravenous lines.

TRPV4 Channel is an Osmosensor and Mechanosensor in Proximal Tubule Cells

The proximal tubule (PT) is the first segment of the nephron and accounts for reabsorption of water and solutes filtered by the glomerulus. In addition, the proximal tubule plays a pivotal role in the endocytosis to reabsorb and metabolize proteins and other substances in glomerular filtrates. Recent studies showed that the PT functions can be modulated by mechanical forces, namely the shear stress on the apical surface and the radial stretch on tubular lumen. Since TRPV4 channel is mainly expressed at the basolateral side of the tubule, we hypothesized that TRPV4 could play a role as stretch-activated channel rather than a shear stress sensor. Patch clamp and Ca2+ imaging experiments showed that TRPV4 channel is expressed in cultured proximal tubule cells and can be activated by the specific agonist GSK1016790A and by hypotonic solution. Moreover, mechanical stretch induced by uniaxial strain in silicone stretchable chambers activated a large Ca2+ influx that was significantly reduced in Trpv4-/- PT cells. Notably, no TRPV4-dependent Ca2+ entry was observed in response to flow-induced cell deformation. Furthermore, we observed that the uptake of FITC-labeled albumin in PT cells, treated with GSK1016790A or mechanically stimulated in stretchable chambers, was significantly reduced in Trpv4-/- PT cells. Finally, we injected angiotensin II in mice, using osmotic minipumps, in order to challenge PT endocytotic activity and increase the proteinuria of mice. Our data showed a massive increase in diuresis of mice. Interestingly, we observed an increase in albuminuria in wild-type mice, and this effect was significantly higher in Trpv4-/- mice. In contrast, glucose and phosphate urinary levels, which undergo PT reabsorption as well, were not significantly affected. Taken together, these results support the view that TRPV4 channel is a mechanosensor in PT cells and modulates their endocytotic activity.

Decellularized human fetal intestine as a bioscaffold for regeneration of the rabbit bladder submucosa

PurposeWe aim to report a method to create a natural acellular scaffold from human fetal small intestine for augmentation cystoplasty in rabbits. MethodsFetal intestines were decellularized by immersion in a hypotonic solution. The success of this protocol was evaluated by histological analysis, scanning electron microscopy and measurement of collagen and sulfated glycosaminoglycan of the acellular tissues. Eight mature rabbits were selected and acellular scaffolds were implanted on the exposed urothelium. Urodynamic studies and cystography were performed after six months. At 14, 120 and 180days animals were sacrificed and augmented bladders were resected. ResultsHistological analysis revealed formation of muscular layer and blood vessels in implanted scaffolds similar to normal bladder. These findings demonstrate the effective seeding of scaffold by host bladder cells. The tissue architecture of recellularized scaffold was similar to the native bladder. ConclusionsFetal intestine acellular matrix could be an exceptional scaffold for bladder augmentation cystoplasty and may pave the road for future studies in order to be used for clinical application.

Effects of fixation on electrophysiology and structure of human jejunal villi.

The villi of human jejunum vary in size and shape during different functional conditions. In the base the lamina propria is isotonic with blood, in the tip hyperosmotic. Here we study electrophysiological and morphological effects of incubation in hypotonic, isotonic, or hypertonic solutions, and to test various isotonic fixatives for microscopy. Samples of jejunal mucosae, obtained during surgery in obese patients, were studied in Ussing chambers where electrical parameters were registered during incubation in Krebs solution at various osmolarities, and during fixation in formaldehyde, glutaraldehyde, or osmium tetroxide (OsO4 ). The same fixatives were used for other jejunal specimens that were fixed directly for light microscopy. Morphometry was carried out to determine size and height of villi, proportion of lamina propria, and surface enlargement due to villi. Ussing chamber incubation in fluids with low osmolarity resulted in increased electrical resistance and epithelial swelling. Opposite results were obtained at high osmolality. Fixation was faster in formaldehyde than in glutaraldehyde or OsO4 . In biopsies processed directly for light microscopy the proportions of lamina propria of the mucosa, and of lamina propria of villi, were significantly larger in biopsies fixed in formaldehyde than after fixation in glutaraldehyde or OsO4 . The villus tips sometimes ended with a bleb with prominent spaces between the epithelial cells. In summary, jejunal villi swell in vitro when exposed to hypotonic solutions, and shrink in hypertonic solutions. Much of the morphological changes occurring during fixation can be related to the physiological hyperosmolar milieu in villus tips.

Recovery of autologous sickle cells by hypotonic wash

Abstract It is important to isolate autologous red blood cells (RBCs) from transfused RBCs in samples from recently transfused patients to ensure that accurate serologic results are obtained. Typically, this isolation can be performed using methods that separate patient reticulocytes from transfused, older donor RBCs. Patients with sickle cell disease (SCD), however, characteristically have RBCs with altered membrane and morphological features, causing their RBCs to take on a sickle-shape appearance different from the biconcave disc-shape appearance of “normal” RBCs. These characteristics enable the use of hypotonic saline solution to lyse normal RBCs while allowing “sickle cells” to remain intact. Because many patients with SCD undergo frequent transfusions to treat their condition, the use of hypotonic saline solution provides a rapid method to obtain autologous RBCs for serologic testing from this patient population using standard laboratory equipment and supplies. Immunohematology 2018;34: 16–18.

Modulation of endoplasmic reticulum stress and the unfolded protein response in cancerous and noncancerous cells.

Objectives:The bio-field array is a device that generates a dielectrophoretic electromagnetic field when placed in a hypotonic saline solution and a direct current of approximately 3 A is applied. It is known that cell physiology is guided by bioelectrical properties, and there is a significant growth inhibition in cancerous (MDA-MB-231) cells that are grown in media that has been reconstituted with the saline that has been exposed to the bio-field array direct current dielectrophoretic electromagnetic field, alternatively there is no growth inhibition noted in noncancerous cells (MCF-10A) when grown in the bio-field array direct current dielectrophoretic electromagnetic field treated versus control media.Methods:To examine the basis for selective growth inhibition in human breast carcinoma, we employed cell death assays, cell cycle assays, microarray analysis and reverse transcription-quantitative polymerase chain reaction.Results:We found a large transcriptional reprogramming in the cell lines and of the genes affected, those involved in endoplasmic reticulum stress and the unfolded protein response pathways showed some of the most dramatic changes. Cancerous cells grown in media that has been reconstituted with a hypotonic saline solution that has been exposed to the bio-field array direct current dielectrophoretic electromagnetic field show a significant and strong upregulation of the apoptotic arms of the unfolded protein response while the noncancerous cells show a decrease in endoplasmic reticulum stress via microarray analyses and reverse transcription-quantitative polymerase chain reaction.Conclusion:The bio-field array shows potential to initiate apoptosis in cancerous cells while relieving cell stress in noncancerous cells in vitro. These studies lay a foundation for nurses to conduct future in vivo models for the possible development of future adjunct treatments in chronic disease.

To find incidence of hyponatremia in patients administered either of hypotonic or isotonic maintenance fluid for 24 hours

Background : Maintenance intravenous fluids are most commonly necessary in hospitalised children who cannot be fed by enteral route,preoperative and postoperative patients.For almost half a century,most of the Paediatricians around the world were using hypotonic solution(0.2% saline in 5% dextrose) for maintenance fluids according to the guidelines set by Holliday and Segar . The wisdom of this approach to intravenous fluid therapy has been questioned recently ,especially in the hospitalised children who have various osmotic and non-osmotic stimuli for vasopressin release, leading to impaired ability to excrete free water along with secondary natriuresis due to suppression of aldosterone release ,thereby increasing the probability of developing clinically symptomatic hyponatremia.We hypothesized that use of isotonic maintenance fluid (0.9% saline in 5% dextrose) would be more safer than hypotonic fluid (0.14% saline in 5% dextrose) in decreasing the incidence of hospital acquired hyponatremia in the sick children at standard maintenance rate. Methods : This open-labelled randomized controlled trial was conducted in the Paediatric emergency of a secondary care hospital.Children in the age group of 0.6 to 12 years who were admitted and anticipated to receive maintenance intravenous fluid for 24 hours were included into the study.Written informed consent was obtained from the parent or guardian of all the patients before participation in the trial.The study protocol was approved by the Institute Ethics Committee.Children with baseline hyponatremia (serum sodium < 135 mEq/L),hypernatremia(serum sodium >145 mEq/L),neurological disorders,acute diarrhoea and dehydration,shock,head trauma,severe acute malnutritrion,those with pre-existing chronic diseases,diabetic ketoacidosis and suspected poisoning were excluded.Children with fluid overload conditions(like cirrhosis, congestive heart failure,acute and chronic renal failure and nephrotic syndrome)or receiving drugs like diuretics,mannitol,vasopressin, carbamazepine which can cause abnormality in the serum sodium were also excluded.Based on findings of Montanana et al,which showed the incidence of hyponatremia in hypotonic group to be 20.6% and in the isotonic group to be 5.1%,with power of 80% and applying 95%confidence interval,a sample size of 125 patients was calculated by web based power/sample size calculator,out of which 63 were randomized to receive isotonic fluid(0.9% saline in 5% dextrose) and 62 for hypotonic fluid(0.14% saline in 5% dextrose)at standard maintenance rate.Both fluids had potassium in the concentration of 20 mEq/L.Randomisation was done by computer generated random number tables. The allocated intervention was kept in sequentially labelled sealed envelopes and was opened after enrolment of the patient.Baseline demographic, hemodynamic characteristics and level of consciousness were noted at presentation.Level of consciousness,heart rate,respiratory rate,blood pressure andSpO2 were monitored every 6 hourly for first 24 hours of admission.Baseline investigations like complete blood counts,serum sodium,serum potassium,blood sugar,renal function tests and those relevant to the patient illness were done. Serum osmolality was calculated at the time of enrolment and after 24 hours by the formula:Serum osmolality=(2 × serum sodium) + (blood sugar/18) + (blood urea/2.8).Serum sodium was repeated after 24 hours of intravenous fluid therapy. All enrolled patients were monitored for signs and symptoms of sodium imbalance,signs of dehydration during first 24 hours. Results : Baseline variables were comparable in two groups. Hyponatremia at 24 hours was seen in 11(17.4%)patients in isotonic and in 29(47%)patients of hypotonic group (P=0.02).The odds ratio for developing hyponatremia in hypotonic group was 4.32(95%CI: 1.89, 9.90)as compared to isotonic group.There were 25 patients (59.5%) of respiratory tract illness who developed hyponatremia in hypotonic group as compared to 6 patients (13.9%)of isotonic group (P=0.001).At 24 hours,both mean serum sodium and mean serum osmolality were significantly lower in hypotonic group as (P=0.002) and (P=0.001) respectively Conclusion : To conclude,the type of fluid is important in deciding the incidence of hyponatremia in hospitalized children. Among the fluids,isotonic fluids are safer choice than the traditional hypotonic fluids in hospitalized children requiring maintenance fluid therapy.Current clinical situations demand such a maintenance solution and volume which maintains tonicity and balance in acute illness,rather than one which merely provides a daily sodium or calorie requirement.

The effect of oral versus intravenous fluid therapy on maternal and neonatal outcomes for women with oligohydramnios

Background Oligohydramnios may increase the perinatal morbidity and mortality rate and is associated with adverse fetal outcomes. Aim The aim of this study was to assess the effect of oral versus intravenous fluid therapy on maternal and neonatal outcomes for women with oligohydramnios. Study design This was a quasi-experimental study model. Setting Th study was conducted at a high-risk pregnancy department at a specific private hospital in El-Mansoura City, Egypt. Participants and methods A purposive sample of 45 pregnant women with oligohydramnios was included in the study, and the women were divided equally into three groups, with 15 women per each group: the first group was administered intravenous (i.v.) isotonic solution, second group was administered i.v. hypotonic solution, and the third group was administered oral water. Data were collected during the period of 6 months from the first of December 2016 to the end of August 2017. Tools Four tools were used to conduct this study: maternal characteristics questionnaire, sonographic report for measuring amniotic fluid index (AFI) in oligohydramnios women before and after hydration, fluid chart, and maternal and neonatal assessment sheet. Results The AFI was significantly increased from 0.35±0.07 to 1.7±0.5 in the i.v. isotonic group whereas the AFI was significantly increased from 0.37±0.08 to 1.9±0.9 in the i.v. hypotonic group. Moreover, the AFI was more markedly increased from 0.37±0.07 to 2.7±0.8 in the oral water group. The maternal and neonatal outcomes did not differ significantly among the groups but were more markedly increased in the oral water group. Conclusion Oral hydration is as effective as intravenous hydration in significantly increasing the AFI in the third trimester oligohydramnios. The AFI was more markedly increased in the oral water. Maternal and neonatal outcomes did not differ significantly among the groups. Recommendations Oral hydration is recommended for pregnant women with oligohydramnios till delivery.

A comparative study of isotonic and hypotonic intravenous maintenance fluids in hospitalised children

Background : For almost half a century,most of the Paediatricians around the world were using hypotonic solution(0.2% saline in 5% dextrose) for maintenance fluids according to the guidelines set by Holliday and Segar .The wisdom of this approach has been questioned recently, especially in the hospitalised children who have various osmotic and non-osmotic stimuli for vasopressin release, leading to clinically symptomatic hyponatremia.We hypothesized that use of isotonic maintenance fluid (0.9% saline in 5% dextrose) would be more safer than hypotonic fluid (0.14% saline in 5% dextrose) in sick children at standard maintenance rate. Methods : This open-labelled randomized controlled trial was conducted on 125 admitted children of secondary care hospital,in the age group of 0.6 to 12 years who were anticipated to receive maintenance intravenous fluid for first 24 hours.63 were randomized to receive isotonic fluid(0.9% saline in 5% dextrose) and 62 for hypotonic fluid(0.14% saline in 5% dextrose)at standard maintenance rate by computer generated tables.Baseline investigation,serum sodium,serum potassium,blood sugar,renal function tests and those relevant to the patient illness were done. Serum osmolality was calculated at the time of enrolment and after 24 hours.Serum sodium was repeated after 24 hours of intravenous fluid therapy. All enrolled patients were monitored for signs and symptoms of sodium imbalance,signs of dehydration during first 24 hours. Results : Baseline variables were comparable in two groups. Hyponatremia at 24 hours was seen in 11(17.4%)patients in isotonic and in 29(47%)patients of hypotonic group (P=0.02).The odds ratio for developing hyponatremia in hypotonic group was 4.32(95%CI: 1.89, 9.90)as compared to isotonic group.There were 25 patients (59.5%) of respiratory tract illness who developed hyponatremia in hypotonic group as compared to 6 patients (13.9%)of isotonic group (P=0.001).At 24 hours,both mean serum sodium and mean serum osmolality were significantly lower in hypotonic group as (P=0.002) and (P=0.001) respectively Conclusion : To conclude,the type of fluid is important in deciding the incidence of hyponatremia in hospitalized children.Among the fluids,isotonic fluids are safer choice than the traditional hypotonic fluids in hospitalized children requiring maintenance fluid therapy.

Сравнительное исследование эффективности применения таутомеров оротата магния для компенсации дефицита магния. Часть I. влияние таутомеров магния оротата на изолированные клетки лабораторных животных и человека

The objects of the research were erythrocytes and lymphocytes of humans and rats and human buccal epitheliocites. The hemolysis of erythrocytes and microelectrophoresic cell mobility in solutions of saline with the orotate magnesium tautomers was studied. The mobility of erythrocytes in the solution with the dihydroxy-form showed negative dynamics (the number of active erythrocytes decreased in 2 times), the activity of cells increased in hydroxy-forms solutions: the amplitude of vibrations of the erythrocyte membrane increased more than in 2 times. Moreover, in solutions with hydroxy- and dihydroxy-forms the active forms of lymphocytes appeared. In oxo-form solution this effect was not observed. Buccal epithelium cells react in a similar way. The amplitude cytolemma and the nuclear membrane vibrations is higher in solution with hydroxy-form magnesium orotate. Activation of nucleated buccal cells in solutions of hydroxy-form are more pronounced than activation of red blood cells. The stability of the erythrocyte membrane of animals with experimental magnesium deficiency to the action of a hypotonic solution is restored on the 10th day after taking tautomers and is more pronounced when taking the hydroxyl-form of magnesium orotate.Magnesium orotate tautomers have different mechanisms of absorption and have different properties in relation to participation in biochemical processes.

Study of Osmotic Fragility Status of Red Blood Cell in Type II Diabetes Mellitus Patients

Diabetes Mellitus (DM) is one of the most prevalent non-communicable diseases in the world. Cell membrane injury is an important mechanism for pathophysoilogical changes in DM. Osmotic fragility (OF) status of Red blood cell (RBC) in hyperglycemic patients is expected to be increased. This study was conducted in Chittagong medical college hospital and Chittagong Diabetic Hospital from January 2015 to December 2015. 100 newly diagnosed (duration ≤ 3 years) type II diabetes mellitus patients (Fasting blood glucose is ≥7 mmol/L) were selected as cases. Age, sex and BMI matched 100 healthy subjects were included as control. OF of RBC was measured by traditional method with a series of hypotonic solution of NaCl of different strength in twelve test tubes numbered serially. The relationship of OF with Fasting blood sugar (FBS) and two hours post prandial blood sugar (2 HPPBS) were evaluated. Unpaired Student ‘t’ test and Pearson’s correlation coefficient test were done for statistical analysis. p value <0.05 was considered as significant. 87% of cases showed normal hemolysis and only 12% had early hemolysis. Mean value of NaCl solution for partial and complete hemolysis in cases were 0.44±0.06(%) and 0.32±0.02(%) respectively; for control group the findings were 0.04±0.06(%) and 0.32±0.02(%). Significant relationship (p<0.001) was found regarding osmotic fragility with FBS ≥ 7 mmol/L and 2 HPPBS ≥ 11.1 mmol/L. OF of RBC is increased in type II diabetes mellitus.

An idiogram on pachytene bivajents with high resalution multiple bands of Epinephelus malabaricus

Epinephelus malabaricus (Perciformes, Serranidae), a rare marine commercial fish inhabits in the warm-water reef along the southeast coast of China. In the present study, well-spread pachytene bivalents with multiple high-resolution bands were obtained after the testes were treated with a hypotonic alkaline solution and a high-chloroform fixative solution, showing that the number and characteristics of band in each bivalent were stable. According to karyotype analysis, the haploid chromosome set was n=24 (24t), and no heteromorphic bivalent was found. The relative length of the longest and shortest bivalent were 5.86 and 1.66, and the length ratio was 3.53, which was much higher than the length ratio of the exact metaphase mitotic karyotype (1.5) and premetaphase endonuclease karyotype (1.75). An idiogram with 447 bands was constructed, and the zone of each bivalent was divided, respectively. The detailed description of the landmark system and the band positions were preliminary given. This study aims to provide a theoretical reference for the further research on cytogenetics and the physical map construction of chromosome fine frame of E. malabaricus.

Angiotensin II Type 1 Receptor Mechanoactivation Involves RGS5 (Regulator of G Protein Signaling 5) in Skeletal Muscle Arteries: Impaired Trafficking of RGS5 in Hypertension.

Studies suggest that arteriolar pressure-induced vasoconstriction can be initiated by GPCRs (G protein-coupled receptors), including the AT1R (angiotensin II type 1 receptor). This raises the question, are such mechanisms regulated by negative feedback? The present studies examined whether RGS (regulators of G protein signaling) proteins in vascular smooth muscle cells are colocalized with the AT1R when activated by mechanical stress or angiotensin II and whether this modulates AT1R-mediated vasoconstriction. To determine whether activation of the AT1R recruits RGS5, an in situ proximity ligation assay was performed in primary cultures of cremaster muscle arteriolar vascular smooth muscle cells treated with angiotensin II or hypotonic solution in the absence or presence of candesartan (an AT1R blocker). Proximity ligation assay results revealed a concentration-dependent increase in trafficking/translocation of RGS5 toward the activated AT1R, which was attenuated by candesartan. In intact arterioles, knockdown of RGS5 enhanced constriction to angiotensin II and augmented myogenic responses to increased intraluminal pressure. Myogenic constriction was attenuated to a higher degree by candesartan in RGS5 siRNA-transfected arterioles, consistent with RGS5 contributing to downregulation of AT1R-mediated signaling. Further, translocation of RGS5 was impaired in vascular smooth muscle cells of spontaneously hypertensive rats. This is consistent with dysregulated (RGS5-mediated) AT1R signaling that could contribute to excessive vasoconstriction in hypertension. In intact vessels, candesartan reduced myogenic vasoconstriction to a greater extent in spontaneously hypertensive rats compared with controls. Collectively, these findings suggest that AT1R activation results in translocation of RGS5 toward the plasma membrane, limiting AT1R-mediated vasoconstriction through its role in Gq/11 protein-dependent signaling.

Preparation of Meiotic Chromosome Spreads from Mouse Spermatocytes.

Mammalian meiosis is a dynamic developmental process that occurs in germ cells and can be studied and characterized. Using a method to spread nuclei on the surface of slides (rather than dropping them from a height), we demonstrate an optimized technique on mouse spermatocytes that was first described in 1997. This method is widely used in laboratories to study mammalian meiosis because it yields a plethora of high quality nuclei undergoing substages of prophase I. Seminiferous tubules are first placed in a hypotonic solution to swell spermatocytes. Then spermatocytes are released into a sucrose solution to create a cell suspension, and nuclei are spread onto fixative-soaked glass slides. Following immunostaining, a diversity of proteins germane to meiotic processes can be examined. For example, proteins of the synaptonemal complex, a tripartite structure that connects the chromosome axes/cores of homologs together can be easily visualized. Meiotic recombination proteins, which are involved in repair of DNA double-strand breaks by homologous recombination, can also be immunostained to evaluate progression of prophase I. Here we describe and demonstrate in detail a technique widely used to study mammalian meiosis in spermatocytes from juvenile or adult male mice.

Proliferative activity of bone marrow cells of rats under the influence of biologically active substances of a medical leech

Hirudotherapy has been used in medicine since time immemorial. The advantage of this type of treatment is that the bite of a leech has a restorative effect on the entire body. The leech bites the skin and injects into the patient’s blood biologically active substances that activate blood microcirculation and promote immunity. The saliva of the leech contains more than 100 substances that contribute to the spectrum of therapeutic effects. But there are no studies on the effect of hirudotherapy on the proliferation of bone marrow cells, which is ontogenetically primary in the development of myeloid and lymphoid tissues. Therefore, it is important to study the mitotic activity of bone marrow of rats at the early stages of postembryonic development against the background of influence by the leech. Female nonlinear rats, two weeks before and two weeks after mating with males, were presented each week to one hungry medical leech weighing 400 mg. The females were examined after feeding their offspring and their offspring were likewise examined on day 1, 15, 30, 45, 60 of the experiment. The animals were kept in vivarium conditions on a standard diet in individual cages. All animals were decapitated under ether anesthesia and the number of leukocytes, the leukocyte blood count, the number of erythrocytes, hemoglobin, and the color index were examined. After that, the femurs were quickly removed and the adjacent muscles separated with a sterile gauze. We cut the epiphyses and cut along the bone. Bone marrow from the femur was flushed with a warm (37 °C) hypotonic 0.9% sodium citrate solution into a sterile centrifuge tube using a Pasteur pipette. The resulting cell suspension was incubated in a hypotonic solution for 10 minutes at 37 °C. The resulting suspension was centrifuged for 5 minutes at 1000 rpm leaving only a small residue of sediment in the test tube. The cells were fixed in methyl alcohol with glacial acetic acid (three parts methyl alcohol and one part acetic acid). 3,000 cells were analyzed, among which we determined those that are in mitosis, and expressed their share. As a result of our studies, the immunomodulating effect of the saliva of the medical leech was revealed, which manifested itself in an increase in the proliferative activity of bone marrow cells, which led to an increase in the number of red blood cells, increased hemoglobin and increase in the number of leukocytes. The color index and the leukocyte blood count were within the normal limits.

Adult exposure to bisphenol A in rare minnow Gobiocypris rarus reduces sperm quality with disruption of testicular aquaporins

Bisphenol A (BPA) is an endocrine disrupter which has adverse effects on male reproduction. Aquaporins (AQPs), well known water-selective channels, play important roles in spermatogenesis and sperm functions. However, whether AQPs participate in the process that BPA induces abnormal sperms has not been investigated to date. In the present study, adult male rare minnows Gobiocypris rarus were exposed to environmentally relevant concentrations BPA (15 and 225 μg/L) for 1, 2 and 3 weeks. Results showed that BPA exposure disrupted sperm motility, increased the percentage of abnormal sperm cells, and decreased sperm tolerance to hypotonic solution and sperm fertilization capacity. Meanwhile, protein levels of AQPs were up-regulated, and their distribution in the testis was abnormal following BPA exposure. The following chromatin immune coprecipitation showed that BPA could regulate aqp3 and 8 expression through the ERE in their 5′-flanking region. The present study demonstrated that BPA could decrease the sperm quality in rare minnow, and AQP3 and 8 might play significant roles in this process.

Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk.

Extracellular matrix (ECM) is an essential component of the tissue microenvironment, actively shaping cellular behavior. In vitro culture systems are often poor in ECM constituents, thus not allowing for naturally occurring cell-ECM interactions. This study reports on a straightforward and efficient method for the generation of ECM scaffolds from lung tissue and its subsequent in vitro application using primary lung cells. Mouse lung tissue was subjected to decellularization with 0.2% sodium dodecyl sulfate, hypotonic solutions, and DNase. Resultant ECM scaffolds were devoid of cells and DNA, whereas lung ECM architecture of alveolar region and blood and airway networks were preserved. Scaffolds were predominantly composed of core ECM and ECM-associated proteins such as collagens I-IV, nephronectin, heparan sulfate proteoglycan core protein, and lysyl oxidase homolog 1, among others. When homogenized and applied as coating substrate, ECM supported the attachment of lung fibroblasts (LFs) in a dose-dependent manner. After ECM characterization and biocompatibility tests, a novel in vitro platform for three-dimensional (3D) matrix repopulation that permits live imaging of cell-ECM interactions was established. Using this system, LFs colonized the ECM scaffolds, displaying a close-to-native morphology in intimate interaction with the ECM fibers, and showed nuclear translocation of the mechanosensor yes-associated protein (YAP), when compared with cells cultured in two dimensions. In conclusion, we developed a 3D-like culture system, by combining an efficient decellularization method with a live-imaging culture platform, to replicate in vitro native lung cell-ECM crosstalk. This is a valuable system that can be easily applied to other organs for ECM-related drug screening, disease modeling, and basic mechanistic studies.

Iatrogenic Dysnatremias in Children with Acute Gastroenteritis in High-Income Countries: A Systematic Review.

Acute gastroenteritis (AGE) causing dehydration with or without dysnatremias is a common childhood health challenge. While it is accepted that oral rehydration therapy is preferred, clinical factors or parent and healthcare provider preferences may lead to intravenous rehydration (IVR). Isotonic solutions are increasingly recommended in most scenarios requiring IVR. Nevertheless, children with AGE, having ongoing losses of water and electrolytes, represent a unique population. To evaluate the association between acquired dysnatremias and IVR in children with AGE. A systematic search of MEDLINE database was conducted through September 14, 2016. Observational studies and clinical trials conducted in high-income countries were included. The Grades of Recommendation, Assessment, Development, and Evaluation approach was used to evaluate the overall quality of evidence for each outcome. 603 papers were identified of which 6 were included (3 randomized controlled trials and 3 observational studies). Pooling of patient data was not possible due to significantly different interventions or exposures. Single studies results demonstrated that within 24 h, administration of isotonic saline was not associated with a significant decline in serum sodium while hypotonic solutions (0.2-0.45% saline) were associated, in one study, with mean serum sodium declines from 1.3 mEq/L (139.2, SD 2.9-137.9, SD 2.5) in 133 young infants (aged 1-28 months), to 5.7 (SD 3.1) mEq/L in a subgroup of 18 older children (age mean 5.8, SD 2.7 years). Both isotonic and hypotonic saline were shown to be associated with improvement of baseline hyponatremia in different studies. Baseline hypernatremia was corrected within 4-24 h in 81/83 (99.6%) children using hypotonic saline IVR. There is a paucity of publications assessing the risk for acquired dysnatremias associated with IVR in children with AGE. Current high-quality evidence suggests that, short-term use of isotonic solutions is safe and effective in most children with AGE; hypotonic solutions may also be appropriate in some subpopulations, however, the quality of available evidence is low to very low. Further research investigating outcomes associated with IVR use beyond 24 h focusing on specific age groups is required.

Simulation of the osmosis-based drug encapsulation in erythrocytes.

Drug-loaded erythrocytes have been proposed for the treatment of disease. A common way to load drugs into erythrocytes is to apply osmotic shock. Currently, osmosis-based drug encapsulation is studied mainly experimentally, whereas a related theoretical model is still incomplete. In this study, a set of equations is developed to simulate the osmosis-based drug-encapsulation process. First, the modeling is validated with hemolysis rates and the drug-loaded quantities to be found in the literature. Then, the variation of the erythrocyte volume, formation of the pore on the erythrocyte membrane, and quantities of drug loaded into and hemoglobin released from erythrocytes are studied. Finally, an optimized operating condition for encapsulating drugs is proposed. The results show that the volume of erythrocytes exposed to hypotonic NaCl solution increases first and then abruptly decreases because of the pore formation; afterwards, it again increases and then decreases slowly. In the presence of the pore, the drug is loaded by diffusion, whereas the leak-induced convection goes against the loading. For an allowed 45% hemolysis rate, with a 10% hematocrit, the optimized NaCl concentration is 0.44%, the optimized time for sealing the loaded erythrocytes with hypertonic NaCl solution is at 6.5s, and the quantity of albumin (drug) loaded is 4.5mg/ml cells.

Blockade of potassium ion transports enhances hypotonicity-induced cytocidal effects in gastric cancer.

BackgroundPeritoneal lavage with distilled water has been used for surgeries of various cancers to reduce peritoneal recurrence. This study examined whether blockade of potassium ion transports enhances hypotonicity-induced cytocidal effects in gastric cancer (GC).ResultsA potassium channel blocker inhibited the occurrence of regulatory volume decrease (RVD) induced by mild hypotonic stimulation, and significantly enhanced cytocidal effects on GC cells. Incubating MKN45 cells with hypotonic solutions containing a potassium channel blocker significantly reduced the formation of peritoneal metastases in nude mice.MethodsThe three human GC cell lines (HGC-27, Kato III, and MKN45) were exposed to mild hypotonic solutions, and the effects of blockade of potassium ion transports during hypotonic stimulation on cell volume changes and cell viabilities were examined. In the in vivo study, MKN45 cells stimulated with mild hypotonic solutions were intraperitoneally injected into nude mice, and the effects of blockade of potassium ion transports during hypotonic stimulation on the formation of peritoneal metastases were evaluated.ConclusionsBlockade of potassium ion transports enhances hypotonicity-induced cytocidal effects on GC cells, which may contribute to development of a novel lavage method for further reduction of peritoneal recurrence in GC.