We hypothesized that progesterone-induced decrease in uterine fluid pH involves V-ATPase. In this study, expression and functional activity of V-ATPase in uterus were investigated under progesterone influence. Ovariectomized adult female rats received subcutaneous injection of estradiol-17β (1 µg/kg/day) or progesterone (20 mg/kg/day) for 3 days or 3 days estradiol-17β followed by 3 days vehicle, progesterone, or estradiol-17β plus progesterone. Mifepristone, a progesterone receptor blocker, was concomitantly given to the rats which received progesterone. A day after last injection, rate of uterine fluid secretion, its HCO3 (-) concentration, and pH were determined via in vivo uterine perfusion in rats under anesthesia. V-ATPase inhibitor, bafilomycin, was introduced into the perfusion buffer, and changes in these parameters were observed. Expression of V-ATPase A1 and B1/2 proteins and mRNAs in uterus were quantified by Western blotting and real-time PCR, respectively. Distribution of these proteins was observed by immunohistochemistry. Our findings showed that under progesterone influence, uterine fluid secretion rate, HCO3 (-) concentration, and pH were significantly reduced. Administration of bafilomycin did not cause significant changes in fluid secretion rate; however, HCO3 (-) concentration and pH were significantly elevated. In parallel with these changes, expression of V-ATPase A1 and B1/2 proteins and mRNAs were significantly increased with these proteins highly distributed in uterine luminal and glandular epithelia. In conclusion, increased expression and functional activity of V-ATPase were most likely responsible for the decreased in uterine fluid pH observed under progesterone influence.
The assessment of moisture loss from meat during the aging period is a critical issue for the meat industry. In this article, a non-invasive microwave ring-resonator sensor is presented to evaluate the moisture content, or more precisely water holding capacity (WHC) of broiler meat over a four-week period. The developed sensor has shown significant changes in its resonance frequency and return loss due to reduction in WHC in the studied duration. The obtained results are also confirmed by physical measurements. Further, these results are evaluated using the Fricke model, which provides a good fit for electric circuit components in biological tissue. Significant changes were observed in membrane integrity, where the corresponding capacitance decreases 30% in the early aging (0D-7D) period. Similarly, the losses associated with intracellular and extracellular fluids exhibit changed up to 42% and 53%, respectively. Ultimately, empirical polynomial models are developed to predict the electrical component values for a better understanding of aging effects. The measured and calculated values are found to be in good agreement.
This paper discusses the application of a reagentless, selective microbiosensor as a useful alternative tool for monitoring D-serine in neural samples. The main components of the 125-μm-diameter disk biosensor were D-amino acid oxidase for D-serine sensitivity (linear region slope, 61 ± 7 μA cm(-2) mM(-1); limit of detection, 20 nM), and poly-phenylenediamine for rejection of electroactive interference. The response time of the biosensor was of the order of 1 s, ideal for 'real-time' monitoring, and detection of systemically administered D-serine in brain extracellular fluid is demonstrated. Exploitation of this probe might resolve queries involving regulation of D-serine in excitotoxicity, and modulation of N-methyl-D-aspartate receptor function by D-serine and glycine in the central nervous system.
Diabetic Macular Edema (DME) is caused by accumulation of extracellular fluid from hyperpermeable capillaries within the macula. DME is one of the leading causes of blindness among Diabetes Mellitus (DM) patients. Early detection followed by laser photocoagulation can save the visual loss. This review discusses various imaging modalities viz. biomicroscopy, Fluorescein Angiography (FA), Optical Coherence Tomography (OCT) and colour fundus photographs used for diagnosis of DME. Various automated DME grading systems using retinal fundus images, associated retinal image processing techniques for fovea, exudate detection and segmentation are presented. We have also compared various imaging modalities and automated screening methods used for DME grading. The reviewed literature indicates that FA and OCT identify DME related changes accurately. FA is an invasive method, which uses fluorescein dye, and OCT is an expensive imaging method compared to fundus photographs. Moreover, using fundus images DME can be identified and automated. DME grading algorithms can be implemented for telescreening. Hence, fundus imaging based DME grading is more suitable and affordable method compared to biomicroscopy, FA, and OCT modalities.
Metabonomics can be used to quantitatively measure dynamic biochemical responses of living organisms to physiological or pathological stimuli. A range of analytical tools such as high-resolution nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) combined with multivariate statistical analysis can be employed to create comprehensive metabolic signatures of biological samples including urine, plasma, faecal water and tissue extracts. These metabolic signatures can reflect the physiological or pathological condition of the organism and indicate imbalances in the homeostatic regulation of tissues and extracellular fluids. This technology has been employed in a diverse range of application areas including investigation of disease mechanisms, diagnosis/prognosis of pathologies, nutritional interventions and drug toxicity. Metabolic profiling is becoming increasingly important in identifying biomarkers of disease progression and drug intervention, and can provide additional information to support or aid the interpretation of genomic and proteomic data. With the new generation of postgenomic technologies, the paradigm in many biological fields has shifted to either top down systems biology approaches, aiming to achieve a general understanding of the global and integrated response of an organism or to bottom up modelling of specific pathways and networks using a priori knowledge based on mining large bodies of literature. Whilst metabolic profiling lends itself to either approach, using it in an exploratory and hypothesis generating capacity clearly allows new mechanisms to be uncovered.
The combination of novel materials with flexible electronic technology may yield new concepts of flexible electronic devices that effectively detect various biological chemicals to facilitate understanding of biological processes and conduct health monitoring. This paper demonstrates single- or multichannel implantable flexible sensors that are surface modified with conductive metal-organic frameworks (MOFs) such as copper-MOF and cobalt-MOF with large surface area, high porosity, and tunable catalysis capability. The sensors can monitor important nutriments such as ascorbicacid, glycine, l-tryptophan (l-Trp), and glucose with detection resolutions of 14.97, 0.71, 4.14, and 54.60 × 10-6 m, respectively. In addition, they offer sensing capability even under extreme deformation and complex surrounding environment with continuous monitoring capability for 20 d due to minimized use of biological active chemicals. Experiments using live cells and animals indicate that the MOF-modified sensors are biologically safe to cells, and can detect l-Trp in blood and interstitial fluid. This work represents the first effort in integrating MOFs with flexible sensors to achieve highly specific and sensitive implantable electrochemical detection and may inspire appearance of more flexible electronic devices with enhanced capability in sensing, energy storage, and catalysis using various properties of MOFs.
The glycoprotein (G) of Nipah virus (NiV) is important for virus infectivity and induction of the protective immunity. In this study, the extra-cellular domain of NiV G protein was fused with hexahistidine residues at its N-terminal end and expressed in Escherichia coli. The expression under transcriptional regulation of T7 promoter yielded insoluble protein aggregates in the form of inclusion bodies. The inclusion bodies were solubilized with 8 M urea and the protein was purified to homogeneity under denaturing conditions using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. The denatured protein was renatured by gradual removal of the urea. Light scattering analysis of the purified protein showed primarily monodispersity. The purified protein showed significant reactivity with the antibodies present in the sera of NiV-infected swine, as demonstrated in Western blot analysis and enzyme-linked immunosorbent assay (ELISA). Taken together, the data indicate the potential usefulness of the purified G protein for structural or functional studies and the development of immunoassay for detection of the NiV antibodies.