The urinary system regulates ion (electrolyte) levels in the plasma by regulating the amount of sodium, potassium, chloride and other ions lost in the urine.
The urinary system regulates blood pH by regulating the number of H+ and bicarbonate ions (HCO3-) lost in the urine. The k idneys work in concert with lungs to regulate the pH in a narrow limits of buffers within body fluids. 1) releasing renin, a hormone that after a series of reactions eventually restricts salt and water loss at the kidneys. If oxygen levels in the blood are low, the kidneys release erythropoietin, a hormone that stimulates the hemocytoblasts (stem cells in the bone marrow) to increase red blood cell formation. Released by posterior pituitary when osmoreceptors detect an increase in plasma osmolality. Creatinine clearance is the amount of creatine in the urine, divided by the concentration in the blood plasma, over time. Glomerular filtration rate can be calculated by measuring any chemical that has a steady level in the blood, and is filtered but neither actively absorbed or excreted by the kidneys. Creatinine is used because it fulfills these requirements (though not perfectly), and it is produced naturally by the body.
The result of this test is an important gauge used in assessing excretory function of the kidneys.
Other methods involve constant infusions of inulin or another compound, to maintain a steady state in the blood. From the kidneys urine flows down the ureters to the bladder propelled by peristaltic contraction of smooth muscle.
There is a long flat segment as the initial increments of urine enter the bladder and then a sudden sharp rise as the micturition reflex is triggered. This material is based upon work supported by the Nursing, Allied Health and Other Health-related Educational Grant Program, a grant program funded with proceeds of the State’s Tobacco Lawsuit Settlement and administered by the Texas Higher Education Coordinating Board. We will be provided with an authorization token (please note: passwords are not shared with us) and will sync your accounts for you. The editor and reviewers' affiliations are the latest provided on their Loop research profiles and may not reflect their situation at time of review. Soluble sugars play an important role in freezing tolerance in both herbaceous and woody plants, functioning in both the reduction of freezing-induced dehydration and the cryoprotection of cellular constituents. Cold or freezing temperatures are a source of abiotic stress that can dramatically inhibit plant growth and development. A number of different analytical platforms have traditionally been employed in carbohydrate analyses. Capillary zone electrophoresis (CZE) provides a potential alternate method by which soluble carbohydrates from plant samples can be quantified.
Glucose, fructose, sucrose, myo-inositol, mannose, xylose, ribose, lactose, arabinose, and galactose were purchased from Sigma-Aldrich (St. Cabernet franc (CF, Vitis vinifera) grapevines were grown at Horticulture Research Unit 2, Ohio Agricultural Research and Development Center (OARDC), Wooster, OH (lat. Frozen leaves were lyophilized using a FreeZone Plus 12 Liter Cascade Console Freeze Dry System (Labconco Corp., Kansas City, MO, USA), and triple ground in a mortar and pestle using liquid nitrogen. For sugar quantification by GC–MS, grape leaf samples were triple ground in liquid nitrogen as described above, with the following modifications. Capillary zone electrophoresis running conditions were modified from those initially published by Rovio et al. Sugars were derivatized for downstream GC–MS analyses as described previously (Streeter and Strimbu, 1998), with some modifications.


The bladder is a balloon-like bag of smooth muscle =detrussor muscle, contraction of which empties bladder during micturition. This means that you will not need to remember your user name and password in the future and you will be able to login with the account you choose to sync, with the click of a button. This page doesn't support Internet Explorer 6, 7 and 8.Please upgrade your browser or activate Google Chrome Frame to improve your experience. The quantification of soluble sugars in plant tissues is, therefore, essential in understanding freezing tolerance. Freezing temperatures can severely damage crop plants, particularly woody perennials which must over-winter in field conditions. In CZE, molecules injected into the capillary-buffer system migrate in an electrophoretic field, in which the rate of movement of individual molecules through the capillary and detector (usually a photo-diode array) is based on their charge-to-mass ratio (Oefner and Chiesa, 1994). Briefly, stock solutions of carbohydrate calibration standards (myo-inositol, galatinol, stachyose, raffinose, sucrose, lactose [used as an I-STD in CZE analysis], trehalose, cellobiose, galactose, glucose, mannose, fructose, arabinose, xylose, and ribose) were diluted with ultra-pure water.
Sugars were detected using a diode array detector (DAD) set to measure absorbance at a wavelength of 270 nm, with a 10 nm band width. Briefly, sugars were extracted in 75% ethanol, transferred into target vials, and dried under nitrogen gas, as described above.
While a number of analytical techniques and methods have been used to quantify sugars, most of these are expensive and time-consuming due to complex sample preparation procedures which require the derivatization of the carbohydrates being analyzed. Unfortunately, however, detection of carbohydrates under neutral pH conditions, which are not chromophores or fluorophores, following separation via HPLC, is challenging as it must rely on simple UV absorbance of the sugar molecules (Montero et al., 2004). Galactinol, ribitol, and trehalose were supplied by Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Extraction solvents used in the course of the study included: 80% ethanol, 75% ethanol, 60% ethanol, and water. Each aliquot of sample was solvent extracted three times (for a total volume of 600 μL per 20 mg tissue), and the extracts were pooled and dried under nitrogen gas. Samples were separated in alkaline electrolyte buffer using a range of separation voltages. Analysis of soluble sugars using capillary zone electrophoresis (CZE) under alkaline conditions with direct UV detection has previously been used to quantify simple sugars in fruit juices.
Specialty fruit producers, and the grape industry in particular, are at the highest risk of suffering economic losses from cold stress and winter damage. Most notably for the analysis of sugars in plant samples, a CZE method to separate simple sugars in fruit juices has recently been developed under alkaline conditions, using direct UV detection of carbohydrates at a wavelength of 270 nm (Rovio et al., 2007). Grape leaves were harvested on October 17, 2014, and immediately flash-frozen in liquid nitrogen.
The CZE was operated using Beckman Coulter 32 Karat software Version 8.0 (Beckman Coulter). Samples were then vortexed for 10 s and incubated in Reacti-Therm Heating Modules (model 18935, Pierce, Rockford, IL, USA) at 70°C for 40 min.
However, it was unclear whether CZE-based methods could be successfully used to quantify the broader range of sugars present in complex plant extracts. However, both RI and ELS detection systems exhibit maximum sensitivity under aqueous conditions (Montero et al., 2004) and do not permit the use of common HPLC solvents, such as acetonitrile.
LOD and LOQ were determined by monitoring UV absorbance peaks and background absorbance for each dilution of the individual sugar standards. Frozen samples were transported to the laboratory on dry ice and then stored at -80°C until analysis.
As previous studies (Rovio et al., 2007) have demonstrated that fructose, sucrose, and glucose can be resolved with CZE using an alkaline buffer system, a series of basic electrolyte buffers were assayed for use as CZE buffers.


Here, we present the development of an optimized CZE method capable of separating and quantifying mono-, di-, and tri-saccharides isolated from plant tissues. The state of Ohio also experienced its greatest loss since 2010 in 2014 due to consecutive, severe freezing events (Dami and Lewis, 2014).
The LOD threshold was defined as the lowest concentration of a sugar generating a peak with a signal-to-noise ratio of greater than 3, and the LOQ as the lowest concentration of a sugar generating a peak with a signal-to-noise ratio of greater than 10. Following these rinses, the separation voltage was raised linearly from 0 to 10 kV over the course of 2 min and maintained at 10 kV for 10 min.
The column was re-conditioned as described above with fresh running buffer after every 10 runs. In order to survive harsh winters, plants increase their freezing tolerance upon exposure to non-freezing temperatures through cold acclimation (Thomashow, 1999). These columns are therefore often optimized only for the separation of a single class of sugars.
To date, neither CZE methods developed to analyze fruit juice samples, nor those developed to analyze other carbohydrate mixtures, have successfully been used to separate the broader range of sugars present in the complex matrix of most plant extracts. For example, columns optimized for the separation of monosaccharides usually do not resolve oligosaccharides well, and vice-versa, making it difficult to design an HPLC method capable of separating the broad range of carbohydrates found in plant tissues.
The conditioning method above was repeated until a stable baseline absorbance (measured using a UV wavelength of 270 nm) and current were both observed.
The optimized CZE method was successfully used to quantify sugars from grape leaves and buds, and is a robust tool for the quantification of plant sugars found in vegetative and woody tissues. RFOs have been proposed to function as osmo- and cryo-protectants during freezing stress, possibly by serving as a water substitute and forming hydrogen bonds with cellular macromolecules (and maintaining protein secondary and tertiary structure); or by interacting with phospholipid headgroups, preserving plasma membrane integrity and cell turgor (Van den Ende, 2013).
Here, we present an optimized CZE method capable of separating and quantifying mono-, di-, tri-, and tetra-saccharides isolated from both herbaceous and woody samples, using grape leaves as a model plant tissue. Standard sugar solutions were prepared by dissolving individual sugars in ultra-pure (liquid chromatography mass spectrometry-grade, 18 MΩ) water (Fisher Scientific, Pittsburgh, PA, USA). Prior to GC–MS analyses, dilutions were subjected to derivatization, as described below. The increased analytical efficiency of this CZE method makes it ideal for use in high-throughput metabolomics studies designed to quantify plant sugars. The CZE method presented here does not require the use of specialized solvents, or derivatization of the samples, and exhibits an analytical efficiency comparable to (or, for some sugars, greater than) GC–MS methods, making it an ideal method for use in high-throughput metabolomics studies designed to quantify the accumulation of plant sugars throughout cold-stress responses.
LOD and LOQ were determined as described in the CZE section above, with the exception that the total ion count (TIC) peaks generated by the mass spectrometer (electron impact detector) for each sugar, rather than UV absorbance peaks, were employed in signal-to-noise calculations. In grape, previous work has demonstrated that increases in the levels of glucose, fructose, sucrose, raffinose, and stachyose present in buds have a strong positive correlation with freezing tolerance (Grant and Dami, 2015).
In these studies, it was observed that raffinose levels in grape buds were higher in cold hardy than in cold sensitive grape cultivars; and that raffinose levels in basal buds were higher than those in apical buds (Grant and Dami, 2015). Interestingly, similar patterns of raffinose accumulation have been observed in alfalfa cultivars undergoing cold stress, indicating that accumulation of RFOs may also play an important role in freezing tolerance in non-woody species (Castonguay et al., 1995). Given the putative involvement of soluble sugars in plant cold stress responses, the development of reliable methods for quantifying soluble sugars in herbaceous and woody plant tissues is essential in understanding freezing tolerance.



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