Isotonic Control Research Articles

Seawater nasal wash to reduce symptom duration and viral load in COVID-19 and upper respiratory tract infections: a randomized controlled multicenter trial

PurposeThe objective was to assess the efficacy of seawater nasal wash on symptom duration, intranasal viral load, household transmission in COVID-19 and URTIs.MethodsThis prospective, randomized, controlled, multicentric, parallel study included 355 mild/moderate COVID-19 and URTI adults with rhinologic symptoms ≤ 48h. Active group performed 4-daily nasal washes with undiluted isotonic seawater versus control group (without nasal wash). Symptoms were self-assessed daily using the WURSS-21 questionnaire for 3 weeks. Viral load was measured by RT-PCR on nasopharyngeal swabs collected on Day 0, Day 5, Day 14 and Day 21. Digital droplet PCR was additionally performed for SARS-CoV-2.ResultsOverall COVID-19 subjects recovered earlier the ability to accomplish daily activities in the active group (– 1.6 day, p = 0.0487) with earlier improvement of taste (– 2 days, p = 0.0404). COVID-19 subjects with severe nasal symptoms at D0 showed the earliest resolution of anosmia (– 5.2 days, p = 0.0281), post-nasal drip (– 4.1 days, p = 0.0102), face pain/heaviness (– 4.5 days, p = 0.0078), headache (– 3.1 days, p = 0.0195), sore throat (– 3.3 days, p = 0.0319), dyspnea (– 3.1 days, p = 0.0195), chest congestion (– 2.8 days, p = 0.0386) and loss of appetite (– 4.5 days, p = 0.0186) with nasal wash. In URTIs subjects, an earlier resolution of rhinorrhea (– 3.5 days, p = 0.0370), post-nasal drip (– 3.7 days, p = 0.0378), and overall sickness (– 4.3 days, p = 0.0248) was reported with nasal wash.Evolution towards more severe COVID-19 was lower in active vs control, with earlier viral load reduction in youngest subjects (≥ 1.5log10 copies/10000 cells at Day 5: 88.9% vs 62.5%, p = 0.0456). In the active group, a lower percentage of SARS-CoV-2 positive household contacts (0–10.7%) was reported vs controls (3.2–16.1%) among subjects with Delta variant (p = 0.0413).ConclusionThis trial showed the efficacy and safety of seawater nasal wash in COVID-19 and URTIs.Trial registrationTrial registry ClinicalTrials.gov: NCT04916639. Registration date: 04.06.2021.

Impact of streptozotocin-induced diabetes on experimental masseter pain in rats.

This study aimed to assess the influence of streptozotocin (STZ)-induced diabetes on the nociceptive behavior evoked by the injection of hypertonic saline (HS) into the masseter muscle of rats. Forty male rats were equally divided into four groups: a) isotonic saline control, which received 0.9% isotonic saline (IS), (Ctrl-IS); b) hypertonic saline control, which received 5% HS (Ctrl-HS); c) STZ-induced diabetic, which received IS, (STZ-IS); d) STZ-induced diabetic, which received HS (STZ-HS). Experimental diabetes was induced by a single intraperitoneal injection of STZ at dose of 60 mg/kg dissolved in 0.1 M citrate buffer, and 100 μL of HS or IS were injected into the left masseter to measure the nociceptive behavior. Later on, muscle RNA was extracted to measure the relative expression of the following cytokines: cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF-α), and interleukins (IL)-1β, -2, -6, and -10. One-way analysis of variance (ANOVA) was applied to the data (p < 0.050). We observed a main effect of group on the nociceptive response (ANOVA: F = 11.60, p < 0.001), where the Ctrl-HS group presented the highest response (p < 0.001). However, nociceptive response was similar among the Ctrl-IS, STZ-IS, and STZ-HS group (p > 0.050). In addition, the highest relative gene expression of TNF-α and IL-6 was found in the masseter of control rats following experimental muscle pain (p < 0.050). In conclusion, the loss of somatosensory function can be observed in deep orofacial tissues of STZ-induced diabetic rats.

The temporal expression of circulating microRNAs after acute experimental pain in humans.

MicroRNAs (miRNAs) can modulate several biological systems, including the pain system. This study aimed to evaluate the temporal expression of circulating miRNAs in the plasma of healthy volunteers as a marker for epigenetic changes before and after an acute, experimental, pain provocation by intramuscular hypertonic saline injection. Twenty volunteers were randomly allocated into two groups and received either hypertonic (pain) or isotonic (control) saline injection in the first dorsal interosseous muscle of their dominant hand. Pain intensity was continuously recorded for 20minutes after injection on a VAS scale from 0 to 100 (0 indicates no pain and 100 the worst imaginable pain). Blood samples were taken at baseline, 30minutes, 3hours, and 24hours post-injection, and plasma was separated. MiRNA extracts were used for RNA sequencing with the Illumina NextSeq platform. MiRNA transcripts were compared between the pain and the no-pain, control group at every time point. Significant differences were considered when folds were >2 and the False Discovery Rate was p< 0.05. After 30minutes, 4 miRNAs were significantly altered in the pain group compared to controls, which increased to 24 after 3 hours and to 42 after 24hours from baseline (p< 0.0001). Two miRNAs were consistently upregulated throughout the experiment. Enrichment analysis showed significant miRNAs involved in brain perception of pain, brain signalling and response to stimuli. This exploratory study is the first to report on the temporal expression of circulating miRNAs after an acute, human experimental muscle pain model. This exploratory study evaluated the temporal profile of circulating miRNAs in the plasma of healthy subjects after acute experimental pain. Several miRNAs were altered in subjects at the times of follow-up after the acute pain model when compared to controls. MiRNAs previously associated with pain processes were altered in the pain group. Our results, by showing the fast and prolonged modifications of miRNA elicited by the acute experimental pain model, add new perspectives to the topic of epigenetics and pain.

Changes in the expression of certain osmosensitive channel and transporter genes in primary hepatocytes of air-breathing catfish, Clarias magur: A strategy to adapt under osmotic stress

Environmental salinity/osmolarity is an important abiotic ecological factor that affects several metabolic activities of aquatic organisms. Air-breathing magur catfish (Clarias magur) frequently encounter a wide range of external osmolarity changes in their natural habitats as well as internal osmolarity due to the absorption of amino acids and other metabolites through the intestine. The major objective of the present investigation was to demonstrate the influences of anisotonicity exposure on the expression of certain isoforms of aquaporin (AQP) channels (aqp7, 8, and 11), taurine transporter (TauT, slc6a6), urea transporter (UT1, slc14a1) and Na+-K+-ATPase (NKA, atp1a1, and atp1b1) transporter genes, and their translated protein products in the primary hepatocytes of magur catfish. These genes are known to play major roles in cellular volume regulation and water homeostasis under osmotic stress. Exposure of primary hepatocytes to hypertonic medium (+40 mOsmol/L) for a period of 48 h resulted in a significant increase in the expression of aqp7, aqp8, aqp11, slc6a6, slc14a1, atp1a1, and atp1b1 genes, as evidenced by a rise in the levels of both mRNAs of all the above-mentioned genes, and also their translated protein products compared to isotonic controls. However, exposure of primary hepatocytes to hypotonic medium (_40 mOsmol/L) for the same period led to a significant decrease in mRNA levels for aqp7, aqp8, aqp11, slc6a6, and slc14a1 genes, and also their translated protein products compared to isotonic controls as a consequence of downregulation of corresponding genes. Interestingly, NKA enzyme activity in primary hepatocytes was found to increase significantly after exposure to both hypertonic and hypotonic media. Hypertonicity induced the genes for α- and β-subunits of this enzyme both at transcriptional and translational levels. In contrast, hypotonicity induced only the atp1a1 gene and the expression of corresponding translated protein without causing any changes in the expression of the atp1b1 gene. Thus, it may be contemplated that osmotic stress, which results due to anisotonicity of the external environment or in the body fluids, leads to differential expression of different osmosensitive channels such as AQPs, and transporters such as TauT, UT1, UT2, and NKA in hepatic cells of magur catfish and work in coordination to defend the osmotically-induced changes of cellular volume. These regulatory responses of channels and transporters could be one of the vital molecular mechanisms that may have evolved in this catfish to defend against the osmotically-induced stresses.

STUDY OF ISOTONICITY AND OCULAR IRRITATION OF CHLORAMPHENICOL IN SITU GEL

Objective: The objective of this study was to find out the isotonicity of chloramphenicol ophthalmic in situ gel and to know the irritating effect of its in the eyes of test animals, so it can be to maximize absorption of the drug in the eye, minimize drug loss before corneal penetration and safe to used.&#x0D; Methods: This study were started by making four aseptic formulations of in situ gel preparations with a comparison of the baseline concentrations of different Poloxamer 407 and HPMC, F1 (5: 0.45), F2 (10: 0.45), F3 (5: 1) and F4 (10: 1). Four aseptic of in situ gel preparations, followed by a qualitative isotonicity test using blood cells to see the comparison between control and test preparations, and ocular irritation test using the draize test method to determine the presence or absence of the irritation.&#x0D; Results: The results obtained from the isotonicity test showed that the four preparations have normal blood cells that similar with isotonic control solution; therefore, it can be said that the preparations have been made isotonic.&#x0D; The results of the ocular irritation test using the draize test method showed for each category, such as cornea, iris, conjunctiva and edema were zero. A zero value on the cornea indicates no ulceration or opacity, and the iris, conjunctiva and edema were normal. Conclusion: Chloramphenicol in situ gel are isotonic and do not cause irritation to the rabbit's eyes, so they are safe to use and the formulation can be used for further research until the final goal is obtained.

Muscle strengthening of knee extensors in hemiparetics: prospective and longitudinal study

Background: Several attempts to reduce spasticity have been directed to stroke sufferers based on the historical view that spasticity is the major determinant of motor dysfunction and that its reduction results in improved function of the affected limb. Paresis is also recognized as a limiting factor in hemiparetic rehabilitation and has a negative impact on self-care, mobility, or up and down stairs due to slow activation of motor units and difficulty in producing adequate amount of muscle strength. In the treatment of spasticity, many therapeutic procedures have been used. Resisted exercises occur when there is load on the body segment distal to the muscle that develops muscle tension, occurs muscle shortening and an external force is overcome. Objective: The aim of this study was to analyze the results of isometric and isotonic strengthen program in the quadriceps muscles of the affected limb of patients with chronic hemiparesis secondary to stroke and to evaluate its effects on spasticity, motor function of the lower limb, balance and mobility. Methods: The participants were divided into three groups: isotonic strengthening, isometric strengthening and control group. It was applied the Ashworth scale, Fugl-Meyer Protocol, Time Up and Go test and Berg Balance Scale. Results: The isometric and isotonic strengthening program presented post-treatment variations, whereby only the isometry group showed improvement in balance. The control group did not show a significant improvement. There was no increase in the muscular tonus of the quadriceps or hamstrings immediately after the treatment. Conclusion: The selective muscle strengthening of the quadriceps resulted in an increase of the motor function, balance and mobility followed by a decrease or maintenance of the muscular tonus of the patients.

Muscle pain induced by hypertonic saline in the knee extensors decreases single-limb isometric time to task failure

PurposeIncreased nociceptive activity and the experience of exercise-induced pain (EIP) may contribute to fatigue during endurance exercise. To investigate this, a pain model that produces pain similar to EIP and decouples its relationship to exercise intensity is required. This study (1) compared the quality of pain caused by a hypertonic saline injection into the vastus lateralis in resting and exercise conditions, and (2) investigated whether this pain contributes to changes in time to task failure.MethodsOn separate days, 18 participants completed a time to task failure at 20% maximal voluntary torque (MVT), a resting hypertonic saline intramuscular injection, and in a further three visits a time to task failure at 10% MVT following injection of isotonic saline, hypertonic saline or a control (no injection).ResultsIn a subset of eligible participants (n = 12), the hypertonic saline combined with 10% MVT produced a qualitative experience of pain (assessed by the McGill Pain Questionnaire) that felt similar to EIP. 10% MVT with hypertonic saline significantly elevated pain intensity in the first 20% of the time to task failure and caused a significantly (P < 0.05) shorter time to task failure (448 ± 240 s) compared with the isotonic saline (605 ± 285 s) and control (514 ± 197 s) conditions.ConclusionThese findings demonstrate that hypertonic saline increases the intensity of pain during exercise, which results in a faster occurrence of exercise-induced fatigue. These results provide important evidence supporting pain as a limiting factor in endurance performance.

Simultaneous Noxious Stimulation of the Human Anterior Temporalis and Masseter Muscles. Part I: Effects on Jaw Movements.

To test the hypotheses that, in comparison to control (isotonic saline), simultaneous noxious stimulation (hypertonic saline) of the masseter and anterior temporalis muscles would result in (1) reductions in amplitude and velocity of jaw movements during standardized open/close jaw movements and during free and standardized chewing and (2) changes in amplitude and velocity of jaw movements that relate to higher levels of negative mood or pain-related thoughts. Standardized open/close and free and standardized chewing were recorded in 15 asymptomatic participants in three blocks: block 1 (baseline), block 2 (during 5% hypertonic or 0.9% isotonic saline infusion into the right masseter and anterior temporalis muscles simultaneously), and block 3 (infusion sequence reversed). The Depression, Anxiety, and Stress Scale (DASS-21) and the Pain Catastrophizing Scale (PCS) were completed by the participants before the experiment, and the PCS was completed after the experiment. The amplitude and velocity of opening and closing movements for each task were compared between blocks (repeated-measures analysis of variance). Spearman rank correlation coefficient was used to explore correlations. Statistical significance was considered to be P < .05. In comparison to isotonic saline control, hypertonic saline resulted in significantly smaller opening and closing amplitudes and lower velocity during closing in free chewing, but no significant effects in the open/close task or standardized chewing. There were significant correlations between PCS scores and amplitude or velocity during isotonic saline and baseline, but not hypertonic saline. The pain-related reduction in amplitude and/or velocity of free chewing is consistent with the Pain Adaptation Model, but the absence of effects on the open/close task and standardized chewing is not. The few significant correlations between psychologic variables and jaw movement may reflect the low scores.

Antidiabetic activity evaluation of Onobrychis species on alloxan-induced diabetic mice

In the current study, four Onobrychis species, O. albiflora Hub.-Mor., O. argyrea Boiss. subsp. argyrea Boiss., O. galegifolia Boiss., and O. tournefortii (Willd.) Desv. were collected from Anatolia to be evaluated for their antidiabetic activities. Methanol water extracts of the aerial parts were used for experiments. An alloxan-induced diabetic mice test model was used. Phytochemical analysis of the tested extracts was investigated using the HPLC method. The highest activity was observed with treatment of O. albiflora aerial part extract. Significant decrements were detected in the blood glucose levels as follows: 180.83±47.48 and 252.83±50.47mg/dL at 100 mg/kg and 200 mg/kg doses of O. albiflora, respectively, when compared to the isotonic saline solution control group, eliciting a blood glucose level of 494.20±27.32. Among the tested standard compounds, rutin and isoquercetin were detected in the examined species. The highest amount of rutin (1.1981±0.0017%) and isoquercetin (0.7318±0.0197%) were found in O. albiflora and O. argyrea subsp. argyrea, respectively. Antidiabetic activities of the tested Onobrychis species seem to indicate a possible correlation with their rutin and isoquercetin contents. Therefore, rutin and isoquercetin may be the antidiabetic compounds that contribute to the antidiabetic activity of the tested Onobrychis species.

Effects of blueberry on hepatic fibrosis and expression of nuclear transcription factor-кB in rats

Objective: To observe the effects of blueberry and nuclear expression of transcription factor-кb (NF-кb) p65 in an experimental rat model of liver fibrosis. Methods: Forty-five Sprague-Dawley rats were randomly divided into isotonic saline control group (A); model group (B); blueberry juice prevention group (C, 15 g/kg); dan-shao-hua-xian capsule prevention group (D, 1 g/kg); and blueberry juice + dan-shao-hua-xian capsule prevention group (E). Rat liver fibrosis model was established by covalent compound carbon tetrachloride (CCl(4)). Each prevention group was given the corresponding dose of blueberry juice or (and) dan-shao-hua-xian capsule, and the rats were sacrificed after 8 weeks. The degree of liver fibrosis was evaluated by hematoxylin and eosin stain. A liver tissue of NF-κBp65 was detected by immunohistochemical method. The NF-κBp65 protein expression of liver tissue and transforming growth factor (TGF) β1 was detected by Western blot. Data of multiple groups were compared by one-way analysis of variance, and rank sum test. Results: Immunohistochemistry detected that TGFβ1 protein was mainly expressed in mesenchymal origin of hepatic stellate cells. The expression level of group A (3.75 ± 1.67) was low, while those of group B (9.00 ± 2.07), C (7.33 ± 1.00), D (6.00 ± 1.51), and E (3.5 ± 1.41) were high. However, the expression level of TGF-β1 protein in hepatic tissues of group B was significantly higher than that of group C, D and E [group E: 3.5 ± 1.41, F = 18.350, P < 0.05]. In addition, group D was higher than group E (P < 0.05). The expression of NF- kappa Bp65 protein was very complex, and the expression patterns in different groups were different (Statistical calculation of experimental data were based on expression of liver cells). Compared with group B (4.37 ± 2.13), the relative expression levels of NF-κBp65 protein in-group A (0.46 ± 0.25), group C (2.76 ± 1.01), group D (2.13 ± 1.51), group E (1.88 ± 0.99) were significantly decreased (F = 27.490, P < 0.05), and the expression trend was consistent with TGFβ1 protein. Western blot detected NF-κBp65 protein in liver tissues of rats. Compared with group A, levels in groups B, C, D and E were significantly increased (F = 96.983, P < 0.05), and groups C, D and E were significantly lower. The E group was significantly lower than the C group (F = 96.983, P < 0.05), and the degree of hepatic fibrosis was lower in each prevention group than in the B group (T = 24.1, P < 0.05). Conclusion: Blueberries have preventive effect on CCl4-induced hepatic fibrosis in rats, and its preventive mechanism may inhibit the expression and activation of NF-κBp65 in hepatocytes, thereby reducing TGFβ1- mediated production or activation.

Expression of Proinflammatory Cytokines Is Upregulated in the Hypothalamic Paraventricular Nucleus of Dahl Salt-Sensitive Hypertensive Rats.

Accumulating evidence indicates that inflammation is implicated in hypertension. However, the role of brain proinflammatory cytokines (PICs) in salt sensitive hypertension remains to be determined. Thus, the objective of this study was to test the hypothesis that high salt (HS) diet increases PICs expression in the paraventricular nucleus (PVN) and leads to PVN neuronal activation. Eight-week-old male Dahl salt sensitive (Dahl S) rats, and age and sex matched normal Sprague Dawley (SD) rats were divided into two groups and fed with either a HS (4% NaCl) or normal salt (NS, 0.4% NaCl) diet for 5 consecutive weeks. HS diet induced hypertension and significantly increased cerebrospinal fluid (CSF) sodium concentration ([Na+]) in Dahl S rats, but not in normal SD rats. In addition, HS diet intake triggered increases in mRNA levels and immunoreactivities of PVN PICs including TNF-α, IL-6, and IL-1β, as well as Fra1, a chronic marker of neuronal activation, in Dahl S rats, but not in SD rats. Next, we investigated whether this increase in the expression of PVN PICs and Fra1 was induced by increased CSF [Na+]. Adult male SD rats were intracerebroventricular (ICV) infused with 8 μl of either hypertonic salt (4 μmol NaCl), mannitol (8 μmol, as osmolarity control), or isotonic salt (0.9% NaCl as vehicle control). Three hours following the ICV infusion, rats were euthanized and their PVN PICs expression was measured. The results showed that central administration of hypertonic saline in SD rats significantly increased the expression of PICs including TNF-α, IL-6, and IL-1β, as well as neuronal activation marker Fra1, compared to isotonic NaCl controls and osmolarity controls. Finally, we tested whether the increase in PICs expression occurred in neurons. Incubation of hypothalamic neurons with 10 mM NaCl in a culture medium for 6 h elicited significant increases in TNF-α, IL-6, and Fra1 mRNA levels. These observations, coupled with the important role of PICs in modulating neuronal activity and stimulating vasopressin release, suggest that HS intake induces an inflammatory state in the PVN, which, may in turn, augments sympathetic nerve activity and vasopressin secretion, contributing to the development of salt sensitive hypertension.

Sodium lactate improves renal microvascular thrombosis compared to sodium bicarbonate and 0.9% NaCl in a porcine model of endotoxic shock: an experimental randomized open label controlled study

BackgroundSodium lactate seemed to improve fluid balance and avoid fluid overload. The objective of this study was to determine if these beneficial effects can be at least partly explained by an improvement in disseminated intravascular coagulation (DIC)-associated renal microvascular thrombosis.MethodsAncillary work of an interventional randomized open label controlled experimental study. Fifteen female “Large White” pigs (2 months old) were challenged with intravenous infusion of E. coli endotoxin. Three groups of five animals were randomly assigned to receive different fluids: a treatment group received sodium lactate 11.2% (SL group); an isotonic control group received 0.9% NaCl (NC group); a hypertonic control group, with the same amount of osmoles and sodium than SL group, received sodium bicarbonate 8.4% (SB group). Glomerular filtration rate (GFR) markers, coagulation and inflammation parameters were measured over a 5-h period. Immediately after euthanasia, kidneys were withdrawn for histological study. Statistical analysis was performed with nonparametric tests and the Dunn correction for multiple comparisons. A p < 0.05 was considered significant.ResultsThe direct immunofluorescence study revealed that the percentage of capillary sections thrombosed in glomerulus were significantly lesser in SL group [5 (0–28) %] compared to NC [64 (43–79) %, p = 0.01] and SB [64 (43–79), p = 0.03] groups. Alterations in platelet count and fibrinogen level occurred earlier and were significantly more pronounced in both control groups compared to SL group (p < 0.05 at 210 and 300 min). The increase in thrombin–antithrombin complexes was significantly higher in NC [754 (367–945) μg/mL; p = 0.03] and SB [463 (249–592) μg/mL; p = 0.03] groups than in SL group [176 (37–265) μg/mL]. At the end of the experiment, creatinine clearance was significantly higher in SL group [55.46 (30.07–67.85) mL/min] compared to NC group [1.52 (0.17–27.67) mL/min, p = 0.03].ConclusionsIn this study, we report that sodium lactate improves DIC-associated renal microvascular thrombosis and preserves GFR. These findings could at least partly explain the better fluid balance observed with sodium lactate infusion.

Bleeding Time using Moringa oleifera (Malunggay) Leaf Extract versus Saline Control in a Rabbit Epistaxis Model: A Randomized Controlled Trial

Objective: To determine bleeding time using Moringa oleifera leaf extract versus saline control in an experimental epistaxis model.&#x0D; Methods: &#x0D; Design: Randomized controlled trial&#x0D; Setting: Tertiary Government Training Hospital&#x0D; Subjects: Ten adult male New Zealand White rabbits were acclimatized for 1 week in a standard environment. One-centimeter long, full-thickness mucosal wounds in the junction of the nasal floor and anterior part of the septum were treated randomly with topical Moringa oleifera extract or colored isotonic saline control in either right or left nasal cavity, one site at a time. The duration of bleeding – time bleeding started to time bleeding stopped -- was recorded in seconds. Data was subjected to a t-test for paired samples.&#x0D; Results: The mean bleeding time for wounds treated with Moringa extract was 53 seconds (range 38-70 secs), versus 159 seconds (range 100-218 secs) for controls. The bleeding time in the former was significantly shorter than in the latter (p = .000019, t-stat = 8.139), with a mean difference of 106 seconds between the two groups.&#x0D; Conclusion: Moringa oleifera leaf extract was associated with significantly shorter bleeding time than saline control in this experimental epistaxis model, and may be worth investigating further as a hemostatic agent for epistaxis.&#x0D; Keywords: epistaxis, malunggay, Moringa oleifera extract, topical hemostatic agent&#x0D;

Comparison of fluid balance and hemodynamic and metabolic effects of sodium lactate versus sodium bicarbonate versus 0.9% NaCl in porcine endotoxic shock: a randomized, open-label, controlled study

BackgroundSodium lactate has been shown to improve hemodynamics and avoid fluid overload. The objective of this study was to confirm a beneficial effect on fluid balance with sodium lactate infusion and to specify whether the advantage of lactate is related to a negative chloride balance, its particular metabolism, or simply its energy load.MethodsThis was an interventional, randomized, open-label, controlled experimental study. Fifteen female “large white” pigs (2 months old) were challenged with intravenous infusion of Escherichia coli endotoxin. Three groups of five animals were randomly assigned to receive different fluids: a treatment group received sodium lactate 11.2% (SL group); an isotonic control group received 0.9% NaCl (NC group); and a hypertonic control group, with the same amount of osmoles and sodium as the SL group, received sodium bicarbonate 8.4% (SB group). In order to provide the same energy load in the three groups, control groups were perfused with an equivalent energy supply. Statistical analysis was performed with non-parametric tests and the Dunn correction for multiple comparisons at p < 0.05.ResultsFluid and chloride balance, hemodynamics, oxygenation markers, and microcirculatory parameters were measured over a 5-h period. Cumulative fluid balance was significantly lower in the SL group (550 (415–800) mL; median (interquartile range)) compared to the NC group (1100 (920–1640) mL, p = 0.01) and the SB group (935 (790–1220) mL, p = 0.03). Hemodynamics, cardiac efficiency, and microcirculation were significantly enhanced in the SL group, resulting in a significant improvement in oxygen delivery (SL group 417 (305–565) mL/min/m2 at 300 min versus the NC (207 (119–272) mL/min/m2, p = 0.01) and the SB (278, (211–315) mL/min/m2, p = 0.03) groups). Oxygenation markers (arterial oxygen partial pressure (PaO2)/inspired oxygen fraction (FiO2), mixed venous oxygen saturation (SvO2), and venoarterial carbon dioxide tension difference (Pv-aCO2) were enhanced with sodium lactate infusion. Chloride balance was equivalent in both hypertonic groups and significantly reduced compared to the NC group.ConclusionSodium lactate infusion improves fluid balance and hemodynamics. The advantage of lactate does not seem to be explained by its energy load or by the induced negative chloride balance with subsequent water movements.

Experimental pain in the groin may refer into the lower abdomen: Implications to clinical assessments.

To investigate the effects of experimental adductor pain on the pain referral pattern, mechanical sensitivity and muscle activity during common clinical tests. Repeated-measures design. In two separate sessions, 15 healthy males received a hypertonic (painful) and isotonic (control) saline injection to either the adductor longus (AL) tendon to produce experimental groin pain or into the rectus femoris (RF) tendon as a painful control. Pain intensity was recorded on a visual analogue scale (VAS) with pain distribution indicated on body maps. Pressure pain thresholds (PPT) were assessed bilaterally in the groin area. Electromyography (EMG) of relevant muscles was recorded during six provocation tests. PPT and EMG assessment were measured before, during and after experimental pain. Hypertonic saline induced higher VAS scores than isotonic saline (p<0.001), and a local pain distribution in 80% of participants. A proximal pain referral to the lower abdominal region in 33% (AL) and 7% (RF) of participants. Experimental pain (AL and RF) did not significantly alter PPT values or the EMG amplitude in groin or trunk muscles during provocation tests when forces were matched with baseline. This study demonstrates that AL tendon pain was distributed locally in the majority of participants but may refer to the lower abdomen. Experimental adductor pain did not significantly alter the mechanical sensitivity or muscle activity patterns.

AB276. SPR-03 In vitro evaluation of hydrostatic pressure on ATP release and Caspase-1 activation in rat urothelial cells

ObjectiveBladder outlet obstruction (BOO) is projected to affect approximately 1.1 billion men and women by 2018. While BOO results from a multitude of etiologies, they are all characterized by chronically elevated storage and voiding pressures, along with inflammation in the bladder tissue, which can lead to bladder fibrosis and overactive bladder symptoms. Previous research in our lab demonstrated that ATP release by rat primary urothelial cells increased in response to elevated pressure (10–15 cmH2O) in vitro. Thus, we hypothesize that the release of ATP by urothelial cells upon exposure to pathological pressures in BOO initiates a cascade of events that includes the formation of the NLRP3 inflammasome and caspase-1 activation. In the present study, we examined the effects of elevated pressure on ATP-purinergic signaling and caspase-1 activation in rat urothelial cells in vitro.MethodsUsing a custom built pressure system, rat urothelial cell line MYP3 cells (1.2×106 cells/well in F-12 K medium) were exposed to pressure conditions that represent both pathological storage and voiding pressures: 15 cmH2O 60 min, 15, 40 and 75 cmH2O 1 min. Cells that were prepared in a similar manner but maintained under atmospheric pressure served as a control. In addition, MYP3 cells that were exposed to a hypotonic condition (240 mOsm) and cells treated with 1.25 mM ATP served as two positive controls. After exposure to pressure, the supernatant media was collected and the extracellular ATP concentration was measured using a luciferin-luciferase assay kit (Life Technologies). Cells were then lysed and intracellular caspase-1 activity was measured using an established method. The pressure experiments (40 cmH2O 1 min) were repeated in the presence of a P2X7 antagonist [100 µM A-438079 (R&D Systems)] to determine the mechanism for pressure-induced caspase-1 activation.ResultsExposure of MYP3 cells to hydrostatic pressure for 1 min at 15, 40 and 75 cmH2O resulted in a 3-, 3.5- and 4.5-fold increase in extracellular ATP levels, respectively, compared to the 0 cmH2O control. When these cells were exposed to a hypotonic solution, ATP release increased by 6-fold compared to the isotonic control. Exposure of MYP3 cells to pressure also resulted in up to 1.3-fold increases in caspase-1 levels, which were similar to the positive controls, indicating caspase-1 levels are maximized after exposure to pathologic pressure conditions.ConclusionsThe significant increase in ATP release indicates that hydrostatic pressure is a good mechanical stimulus to model BOO in vitro. The acute caspase-1 responses after 1-minute exposure to pressure suggest that high-pressure voiding may be an important trigger of the NLRP3 inflammasome in BOO. The results that demonstrate exposure to pressure and treatment with ATP both result in caspase-1 activation implicate autocrine purinergic signaling as a mediator of NLRP inflammasome formation.Funding Source(s)NIH (R01DK103534, P20GM103444), NSF (1264579)

Reduced Maximal Force during Acute Anterior Knee Pain Is Associated with Deficits in Voluntary Muscle Activation.

Although maximal voluntary contraction (MVC) force is reduced during pain, studies using interpolated twitch show no consistent reduction of voluntary muscle drive. The present study aimed to test if the reduction in MVC force during acute experimental pain could be explained by increased activation of antagonist muscles, weak voluntary activation at baseline, or changes in force direction. Twenty-two healthy volunteers performed maximal voluntary isometric knee extensions before, during, and after the effects of hypertonic (pain) and isotonic (control) saline injections into the infrapatellar fat pad. The MVC force, voluntary activation, electromyographic (EMG) activity of agonist, antagonist, and auxiliary (hip) muscles, and pain cognition and anxiety scores were recorded. MVC force was 9.3% lower during pain than baseline (p < 0.001), but there was no systematic change in voluntary activation. Reduced MVC force during pain was variable between participants (SD: 14%), and was correlated with reduced voluntary activation (r = 0.90), baseline voluntary activation (r = − 0.62), and reduced EMG amplitude of agonist and antagonist muscles (all r > 0.52), but not with changes in force direction, pain or anxiety scores. Hence, reduced MVC force during acute pain was mainly explained by deficits in maximal voluntary drive.

Differential responses of the vasotocin 1a receptor (V1aR) and osmoreceptors to immobilization and osmotic stress in sensory circumventricular organs of the chicken (Gallus gallus) brain

Past studies have shown that the avian vasotocin 1a receptor (V1aR) is involved in immobilization stress. It is not known whether the receptor functions in osmotic stress, and if sensory circumventricular organs may be involved. An experiment was designed with four treatment groups including a 1h immobilization acute stress (AS) group, an unstressed acute control (AC), a third given an intraperitoneal (ip) hypertonic saline injection (HS) and isotonic saline controls (IC) administered ip. One set of chick brains was perfused for immunohistochemistry while a second was sampled for quantitative RT-PCR. Plasma corticosterone (CORT) and arginine vasotocin (AVT) concentrations were significantly increased in the immobilized and hypertonic saline groups (p<0.01) compared to controls. Intense staining of the V1aR occurred throughout the organum vasculosum of the lamina terminalis (OVLT) and subseptal organ (SSO)/subfornical organ (SFO). The immunostaining allowed the boundaries of the two circumventricular organs (CVOs) to be described for the first time in avian species. Both treatment groups showed marked morphological changes in glia within the OVLT and SSO/SFO. The avian V1aR, angiotensin II type 1 receptor (AT1R), and transient receptor potential vanilloid receptor 1 (TRPV1) mRNA levels were increased in the SSO/SFO in hypertonic saline treated birds compared to isotonic controls. In contrast, the latter two genes (AT1R and TRPV1) were significantly decreased in the OVLT of birds subjected to hyperosmotic stress, while all three genes were significantly up-regulated after immobilization. Taken together, results show a possible differential function for the same receptors in two anatomically adjacent CVOs.

Perbandingan pemberian air kelapa muda (cocos nucifera l) dengan isotonik terhadap denyut nadi dan VO2maks atlet remaja

Background: Football players requires a high level of VO2max which is influenced by the available of energy. Tender coconut water and branded isotonic are as energy drinks containing of carbohydrates and the body’s electrolyte ions that can prevent dehydration. The increasing pulse describes a the condition of dehydrated players during match or training. Objective: To Examine comparison of tender hybrid coconut water and branded isotonic to pulse and the plyer’s level of VO2max in the Diponegoro University Football School in Tembalang, Semarang. Methods: Experimental study with an approach of randomized pretest - post test control group design at 41 soccer’s player aged 13-15 years at Diponegoro University Football School Tembalang in August 2015. The treatment group consumed consumed 250 mL tender coconut water (n=13) and branded isotonic (n=14) and group control consumed mineral water (n=14) every 15-30 minutes for 5 times. Observation pre test and post test 1,2,3 with interval of 3 day after physical training, athlets test the pulse to palpation techniques for 15 seconds and VO2max test with multistage fitness test. Result: The VO2max and pulse recovery in the beginning of research for 3 group was similar (p = 0,454 and p = 0,294). Post test 1 VO2max on three group are same (p = 0,841) but the recovery pulse is better on the group of tender coconut water and branded isotonic (p = 0.006). Post test 2, VO2max and recovery pulse is more good by players who were given tender hybrid coconut water and branded isotonic (p = 0,004 and p = 0,017), as the same post test 3 VO2max and recovery pulse (p = 0.000 and p = 0,004).Conclusion: Giving the tender hybrid coconut water and branded isotonic to same to recovery pulse and increased VO2maks on athlete school football diponegoro unversity

Placebo Analgesia Changes Alpha Oscillations Induced by Tonic Muscle Pain: EEG Frequency Analysis Including Data during Pain Evaluation.

Placebo exhibits beneficial effects on pain perception in human experimental studies. Most of these studies demonstrate that placebo significantly decreased neural activities in pain modulatory brain regions and pain-evoked potentials. This study examined placebo analgesia-related effects on spontaneous brain oscillations. We examined placebo effects on four order-fixed 20-min conditions in two sessions: isotonic saline-induced control conditions (with/without placebo) followed by hypertonic saline-induced tonic muscle pain conditions (with/without placebo) in 19 subjects using continuous electroencephalography (EEG) recording. Placebo treatment exerted significant analgesic effects in 14 placebo responders, as subjective intensity of pain perception decreased. Frequency analyses were performed on whole continuous EEG data, data during pain perception rating and data after rating. The results in the first two cases revealed that placebo induced significant increases and a trend toward significant increases in the amplitude of alpha oscillation during tonic muscle pain compared to control conditions in frontal-central regions of the brain, respectively. Placebo-induced decreases in the subjective intensity of pain perception significantly and positively correlated with the increases in the amplitude of alpha oscillations during pain conditions. In conclusion, the modulation effect of placebo treatment was captured when the pain perception evaluating period was included. The strong correlation between the placebo effect on reported pain perception and alpha amplitude suggest that alpha oscillations in frontal-central regions serve as a cortical oscillatory basis of the placebo effect on tonic muscle pain. These results provide important evidence for the investigation of objective indicators of the placebo effect.