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Cureus Dec 2022The bladder is both an intraperitoneal and extraperitoneal structure. Its anatomical position increases its risk of rupture. The resultant urine leak or extravasation...
The bladder is both an intraperitoneal and extraperitoneal structure. Its anatomical position increases its risk of rupture. The resultant urine leak or extravasation can be intraperitoneal, extraperitoneal, or even both-with the former leading to more sinister outcomes. Intraperitoneal bladder rupture can lead to urinary ascites which along with anuria and abdominal pain, can present with an apparent abrupt decline in renal function as the creatinine-rich products diffuse across the peritoneal membrane. Glomerular filtration rate, a measure of kidney function is related to the levels of serum creatinine. Clinicians can therefore misdiagnose their patient with acute kidney injury when the serum creatinine is elevated as a consequence of urine being present in the peritoneal space. This is a case report of a 62-year-old male with pseudo-renal failure following intraperitoneal bladder rupture after a fall face-forwards three hours previously. The fall was due to icy conditions outside and no preceding symptoms were reported. He presented to the Accident and Emergency department with abdominal pain and no other positive symptoms. The patient had a good World Health Organisation (WHO) performance status with a background of hypertension, diabetes, and hypercholesterolemia. The bedside examination of the patient revealed a distended, abdomen with peritonitis. There were no signs of urogenital trauma. Blood testing revealed a low estimated glomerular filtration rate (eGFR) and raised creatinine (eGFR of 7 millilitres/minute and creatinine of 658 micromoles/litre). Computerised tomography examination of the abdomen and pelvis (CTAP) revealed free fluid within the peritoneal cavity and an irregular bladder wall. A CT cystogram and consultation with urology led to the diagnosis of intraperitoneal bladder rupture. The patient's renal function from an initial set of blood tests was reduced. This was not a true impairment in renal function but rather a complication secondary to extravasation of urine in the intraperitoneal space, ie., pseudo renal failure. This supposed impairment in renal function had numerous implications. It affected the choice of antibiotics; amoxicillin and gentamicin were given at a reduced dose due to the patient's renal function and the patient was prepared for operation theatre. The patient's blood creatinine was falsely elevated at 658 micromoles/litre due to the diffusion of creatinine from the free urine in the peritoneal space into the blood. This painted a false image of renal failure and protracted the clinical decision-making process. Relatively simple measures like an ascitic tap could have helped to differentiate this from a true acute kidney injury and could have resulted in quicker and more effective treatment of this patient. The patient went on to have bladder repair under urology. His follow-up cystogram four weeks post-operation did not show any leak.
PubMed: 36514699
DOI: 10.7759/cureus.32360 -
American Journal of Obstetrics and... Jun 2024The principal fetal energy source is glucose provided by the placental transfer of maternal glucose. However, the placenta's glucose consumption exhibits considerable...
BACKGROUND
The principal fetal energy source is glucose provided by the placental transfer of maternal glucose. However, the placenta's glucose consumption exhibits considerable variation. Hexokinase is the first and one of the rate-limiting enzymes of glycolysis that phosphorylates glucose to glucose-6-phosphate. The role of placental hexokinase activity in human placental glucose metabolism is unknown.
OBJECTIVE
This study aimed to test the hypothesis that placental hexokinase activity is related to maternal body mass index, placental glucose uptake and consumption, and birthweight.
STUDY DESIGN
Overall, 67 healthy pregnant participants at term were included in this study at Oslo University Hospital, Oslo, Norway. Placental hexokinase activity was measured by using a colorimetric assay. The mass of glucose taken up by the uteroplacental unit and the fetus was obtained by measuring arteriovenous glucose differences combined with Doppler assessment of uterine and umbilical blood flow. Blood samples were obtained from the maternal radial artery, uterine vein, and umbilical artery and vein. The uteroplacental glucose consumption constituted the difference between uteroplacental and fetal glucose uptakes. The Spearman rank correlation was performed for statistical analyses to study the correlation of placental hexokinase activity (milliunit per milligram of protein) with prepregnancy body mass index, maternal glucose and insulin, birthweight, uteroplacental glucose uptake and consumption, and fetal glucose uptake (micromole per minute). Partial rank correlation analysis was performed when controlling for hours of fasting or placental weight.
RESULTS
Hexokinase activity was detectable in all placental tissue samples. The mean activity was 19.6 (standard deviation, 4.64) mU/mg protein. Placental hexokinase activity correlated positively with prepregnancy body mass index (Spearman rho=0.33; P=.006). On controlling for hours of fasting, hexokinase activity showed positive correlations with both maternal glucose (r=0.30; P=.01) and insulin (r=0.28; P=.02). Hexokinase activity was positively correlated with uteroplacental glucose uptake (Spearman rho=0.31; P=.01) and consumption (Spearman rho=0.28; P=.02). Hexokinase activity did not correlate with fetal glucose uptake. On controlling for placental weight, hexokinase activity showed a positive correlation with birthweight (r=0.31; P=.01).
CONCLUSION
Our findings suggest that placental hexokinase, being crucial for uteroplacental retention of glucose for disposition, is related to both maternal body mass index and birthweight independent of placental weight. Placental hexokinase may play a central role in the relationship between maternal glucose dysregulation and fetal growth. Thus, the current study supports the need to develop clinically useful tools to assess the metabolic properties of the placenta.
Topics: Humans; Female; Hexokinase; Pregnancy; Placenta; Body Mass Index; Birth Weight; Adult; Glucose; Blood Glucose; Infant, Newborn
PubMed: 37925123
DOI: 10.1016/j.ajog.2023.10.043 -
Journal of Investigative Surgery : the... Oct 2021To investigate the relationship of cholelithiasis and urolithiasis with (MTHFR) polymorphism(s) in patients with poor obstetric history to search whether they are risk...
AIM
To investigate the relationship of cholelithiasis and urolithiasis with (MTHFR) polymorphism(s) in patients with poor obstetric history to search whether they are risk factors for adverse pregnancy outcome.
MATERIALS AND METHOD
This study is consisted of 94 patients with poor obstetric history. Patients were evaluated in terms of the presence of cholelithiasis and urolithiasis in association with MTHFR polymorphism(s). Additional laboratory tests including homocysteine measurements were also performed. ROC analysis for assessing the performance of blood homocysteine level in predicting the presence of cholelithiasis and urolithiasis were also performed.
RESULTS
Patients were divided into three groups such as cholelithiasis group (n = 9, 9.6%), urolithiasis group (n = 18, 19.1%) and control group (n = 67, 71.3%). Groups did not differ in term of age and Beksac obstetrics index (BOI) which is "[living child+(π/10)]/gravidity." The rate of the presence of MTHFR polymorphisms were 88.9% (8/9), 88.9% (16/18) and 43.3% (29/67) in cholelithiasis, urolithiasis and control groups respectively. Median homocysteine levels were found to be 13.1, 11.6 and 7.2 micromol/lt for the groups respectively. Statistically significant differences were found for MTHFR polymorphism rates and homocysteine levels (<0.001 for both). According to ROC analysis, 10.9 mcmol/L (88.9% sensitivity, 89.6% specificity) and 9.25 mcmol/L (83.3% sensitivity, 73.1% specificity) were determined to be cutoff values of homocysteine for cholelithiasis and urolithiasis respectively.
CONCLUSION
More frequent MTHFR polymorphisms are observed in women with a clinical history of gall or renal stones. Thus, screening of these patients may be benefical for the approprate management of their subsequent pregnancies.
Topics: Child; Cholelithiasis; Female; Genotype; Humans; Methylenetetrahydrofolate Reductase (NADPH2); Polymorphism, Genetic; Pregnancy; Risk Factors; Urolithiasis
PubMed: 32228104
DOI: 10.1080/08941939.2020.1742402 -
Translational Andrology and Urology Aug 2020Renal cell carcinoma (RCC) is one of the most common malignant tumors in the world. Vitexin (apigenin-8-C-D-glucopyranoside), a bioactive compound isolated from a...
BACKGROUND
Renal cell carcinoma (RCC) is one of the most common malignant tumors in the world. Vitexin (apigenin-8-C-D-glucopyranoside), a bioactive compound isolated from a variety of plants, has multiple protective effects on human health. The purpose of this study was to investigate the role of vitexin in RC and the related molecular mechanism.
METHODS
Proliferation was tested with Cell Counting Kit-8 and Edu staining. Apoptosis was studied with flow cytometry. Immunofluorescent was applied to show LC3 spots. BALB/c nude mice bearing ACHN cells were established and immunohistochemical staining was applied to validate the in vivo effects of vitexin. All the effects and possible signaling pathways involved were validated with western blotting.
RESULTS
Seventy micromole of vitexin started to show significant effect on the growth of normal renal tubular epithelial cells (HK-2), so 0, 10, 20 and 40 µM of vitexin were used in later experiments. Vitexin inhibited growth and induced apoptosis of ACHN and OS-RC-2 cells in a dose-dependent manner, and promoted excessive autophagy by reducing p62 levels and increasing Beclin1 and LC3II levels. Western blotting revealed that vitexin significantly increased the phosphorylation levels of Adenosine Monophosphate Activated Protein Kinase (AMPK) and c-Jun N-terminal kinase (JNK) in ACHN and OS-RC-2 cells, while decreasing the phosphorylation levels of phosphatidylinositol 3-kinase/activates protein kinase/mammalian target of rapamycin (PI3K/AKT/mTOR). In BALB/c nude mice bearing ACHN cells, vitexin inhibited tumor growth, reduced Ki67 and increased caspase-3 levels in the tumor tissues.
CONCLUSIONS
The results indicated that the tumor suppressive role of vitexin in ACHN and OS-RC-2 cells involved AMPK/mTOR, PI3K/AKT/mTOR, and JNK pathways. Therefore, vitexin may be a promising drug for the treatment of RCC.
PubMed: 32944531
DOI: 10.21037/tau-20-1094 -
Molecular Pharmaceutics Feb 2020Magic angle spinning (MAS) NMR is a powerful method for the study of pharmaceutical compounds, and probes with spinning frequencies above 100 kHz enable an...
Magic angle spinning (MAS) NMR is a powerful method for the study of pharmaceutical compounds, and probes with spinning frequencies above 100 kHz enable an atomic-resolution analysis of sub-micromole quantities of fully protonated solids. Here, we present an ultrafast NMR crystallography approach for structural characterization of organic solids at MAS frequencies of 100-111 kHz. We assess the efficiency of H-detected experiments in the solid state and demonstrate the utility of 2D and 3D homo- and heteronuclear correlation spectra for resonance assignments. These experiments are demonstrated for an amino acid, U-C,N-histidine, and also for the significantly larger, natural product Posaconazole, an antifungal compound investigated at natural abundance. Our results illustrate the power for characterizing organic molecules, enabled by exploiting the increased H resolution and sensitivity at MAS frequencies above 100 kHz.
Topics: Antifungal Agents; Carbon Isotopes; Crystallography, X-Ray; Histidine; Hydrogen; Hydrogen Bonding; Magnetic Resonance Imaging; Nitrogen Isotopes; Proton Magnetic Resonance Spectroscopy; Protons; Triazoles
PubMed: 31891271
DOI: 10.1021/acs.molpharmaceut.9b01157 -
BioTechniques Feb 2024Hydrogen sulfide (HS), an endogenous gasotransmitter, has potential applications in several conditions. However, its quantification in simulated physiological solutions...
Hydrogen sulfide (HS), an endogenous gasotransmitter, has potential applications in several conditions. However, its quantification in simulated physiological solutions is a major challenge due to its gaseous nature and other physicochemical properties. This study was designed to compare four commonly used HS detection and quantification methods in aqueous solutions. The four techniques compared were one colorimetric, one chromatographic and two electrochemical methods. Colorimetric and chromatographic methods quantified HS in millimolar and micromole ranges, respectively. The electrochemical methods quantified HS in the nanomole and picomole ranges and were less time-consuming. The HS quantification method should be selected based on the specific requirements of a research project in terms of sensitivity, response time and cost-effectiveness.
Topics: Hydrogen Sulfide; Colorimetry; Spectrum Analysis; Electrochemical Techniques
PubMed: 38059376
DOI: 10.2144/btn-2023-0075 -
International Journal of Environmental... Nov 2022We investigated vanadium, i.e., a redox-active heavy metal widely known for the generation of oxidative stress in cultured mammalian cells, to determine its ability to...
We investigated vanadium, i.e., a redox-active heavy metal widely known for the generation of oxidative stress in cultured mammalian cells, to determine its ability to interfere with common oxidative stress-related bioassays in cell-free conditions. We first assessed the prooxidant abilities (HO level, oxidation of DHR 123, and DCFH-DA dyes) and antioxidant capacity (ABTS, RP, OH, and DPPH methods) of popular mammalian cell culture media, i.e., Minimal Essential Medium (MEM), Dulbecco's Minimal Essential Medium (DMEM), Dulbecco's Minimal Essential Medium-F12 (DMEM/F12), and RPMI 1640. Out of the four media studied, DMEM has the highest prooxidant and antioxidant properties, which is associated with the highest concentration of prooxidant and antioxidant nutrients in its formulation. The studied vanadium compounds, vanadyl sulphate (VOSO), or sodium metavanadate (NaVO) (100, 500, and 1000 µM), either slightly increased or decreased the level of HO in the studied culture media. However, these changes were in the range of a few micromoles, and they should rather not interfere with the cytotoxic effect of vanadium on cells. However, the tested vanadium compounds significantly stimulated the oxidation of DCFH-DA and DHR123 in a cell-independent manner. The type of the culture media and their pro-oxidant and antioxidant abilities did not affect the intensity of oxidation of these dyes by vanadium, whereas the vanadium compound type was important, as VOSO stimulated DCFH-DA and DHR oxidation much more potently than NaVO. Such interactions of vanadium with these probes may artefactually contribute to the oxidation of these dyes by reactive oxygen species induced by vanadium in cells.
Topics: Animals; Reactive Oxygen Species; Vanadium; Antioxidants; Hydrogen Peroxide; Artifacts; Vanadium Compounds; Culture Media; Coloring Agents; Mammals
PubMed: 36429933
DOI: 10.3390/ijerph192215214 -
SLAS Discovery : Advancing Life... Jun 2021A key activity in small-molecule drug discovery is the characterization of compound-target interactions. Surface plasmon resonance (SPR) is a flexible technique for this...
A key activity in small-molecule drug discovery is the characterization of compound-target interactions. Surface plasmon resonance (SPR) is a flexible technique for this purpose, with a wide affinity range (micromoles to picomoles), low protein requirements, and the ability to characterize the kinetics of compound binding. However, a key requirement of SPR is the immobilization of the target protein to the surface of the sensor chip. The most commonly used immobilization techniques (covalent immobilization, streptavidin-biotin) are irreversible in nature, which can afford excellent baseline stability but impose limitations throughput for slowly dissociating compounds or unstable targets. Reversible immobilization (e.g., His-tag-Ni-NTA) is possible but typically precludes accurate quantification of slow dissociation kinetics due to baseline drift.Here we present our investigation of three immobilization strategies (dual-His-tagged target protein, His-tagged streptavidin, and switchavidin) that combine the robustness of irreversible immobilization with the flexibility of reversible immobilization. Each has its own advantages and limitations, and while a universal immobilization procedure remains to be found, these strategies add to the immobilization toolbox that enables previously out-of-scope applications. Such applications are highlighted in two examples that greatly increased throughput for the kinetic characterization of potent kinase inhibitors and kinetic profiling of covalent inhibitors.
Topics: Biosensing Techniques; Drug Discovery; Humans; Kinetics; Small Molecule Libraries; Surface Plasmon Resonance
PubMed: 33289457
DOI: 10.1177/2472555220975358 -
Spectrochimica Acta. Part A, Molecular... Oct 2019Many endogenous peptides are circulating in bodily fluids at micromole level, and accurate analysis of endogenous peptides at such low level is important. In this study,...
Many endogenous peptides are circulating in bodily fluids at micromole level, and accurate analysis of endogenous peptides at such low level is important. In this study, we presented an extensible, facile and sensitive sensor array based on UV-Vis spectroscopy of the AuNPs combined with chemometric methods for quantitative analysis of binary peptide mixture (Gly-Gly/Ala-Gln) using UV-Vis spectroscopy. High concentration arginine (Arg) and Cr can induce aggregation of the AuNPs and DNA-AuNPs. However, the glycylglycine (Gly-Gly) and alanyl-glutamine (Ala-Gln) can prevent the AuNPs from aggregation. We investigated the prevention of AuNPs aggregation by using Gly-Gly and Ala-Gln mixtures and constructed sensor arrays for quantitative analyses of Gly-Gly and Ala-Gln mixtures. The color change of the solution is relevant to the dose of the target, and it can be visualized by the naked eyes or monitored by UV-Vis spectrometry. Results showed that the concentrations of Arg and Cr are the key factors affecting the sensitivity of the sensor array. Whereas when Gly-Gly and Ala-Gln have to be analyzed simultaneously, concentrations of Arg and Cr both for Gly-Gly and Ala-Gln are difficult to be optimized. Taking the advantages of multivariate analysis and data fusion, PLS models and backward interval PLS (BiPLS) models were built for fused dataset constructed by UV-Vis data obtained at different concentrations of Arg and Cr. The best results were obtained from the PLS models. The proposed method can be extended to analysis of other peptides in more complex mixture systems.
Topics: Arginine; Calibration; Chromium; Dipeptides; Glycylglycine; Gold; Humans; Least-Squares Analysis; Limit of Detection; Metal Nanoparticles; Microscopy, Electron, Transmission; Models, Statistical; Reproducibility of Results; Saliva; Signal Processing, Computer-Assisted; Spectrophotometry, Ultraviolet
PubMed: 31158767
DOI: 10.1016/j.saa.2019.117205 -
Bi-enzyme competition based on ZIF-67 co-immobilization for real-time monitoring of exocellular ATP.Mikrochimica Acta Jan 2023Monitoring exocellular adenosine-5'-triphosphate (ATP) is a demanding task but the biosensor development is limited by the low concentration and rapid degradation of...
Monitoring exocellular adenosine-5'-triphosphate (ATP) is a demanding task but the biosensor development is limited by the low concentration and rapid degradation of ATP. Herein, we developed a simple yet effective biosensor based on ZIF-67 loaded with bi-enzymes of glucose (GOx) and hexokinase (HEX) for effective detection of ATP. In the confined space of the porous matrix, the bi-enzymes competed for the glucose substrate in the presence of ATP, facilitating the biosensor to detect low ATP concentrations down to the micromole level (3.75 μM) at working potential of 0.55 V (vs. Ag/AgCl). Furthermore, ZIF-67 with cobalt served as a porous matrix to specifically adsorb ATP molecules, allowing it to differentiate isomers with sensitivity of 0.53 nA/μM, RSD of 5.4%, and recovery rate of 93.3%. We successfully applied the fabricated biosensor to measure ATP secreted from rat PC12 cells in the pericellular space thus realizing time-resolving measurement. This work paved the path for real-time monitoring of ATP released by cells, which will aid in understanding tumor cell glycolysis and immune responses.
Topics: Animals; Rats; Enzymes, Immobilized; Glucose Oxidase; Hexokinase; Adenosine Triphosphate; Glucose
PubMed: 36695915
DOI: 10.1007/s00604-023-05652-y