For the human kidney tissue (glomeruli or tubuli), purified total RNAs were amplified using the Two-Cycle Target Labeling Kit (Affymetrix), as per the manufacturer’s protocol (19 (link)). The raw and analyzed data files have been uploaded to the National Center for Biotechnology Information Gene Expression Omnibus and can be accessed under the following accession numbers: GSE 30122 (all data samples).
Kidney Glomerulus
The kidney glomerulus is the fundamental filtration unit of the kidneys, responsible for removing waste, toxins, and excess water from the blood.
It consists of a network of capillaries surrounded by a specialized epithelial structure called the Bowman's capsule.
The glomerulus plays a crucial role in maintaining fluid balance, electrolyte homeostasis, and blood pressure regulation.
Accurate understanding of glomerular structure and function is essential for research on kidney disease, hypertension, and other related conditions.
PubCompare.ai's AI-driven platform can help optimize your kidney glomerulus research by identifying the best protocols and products through comprehensive comparisons of literature, preprints, and patents, enhancing reproducibility and accurracy with cutting-edg tools.
Start your research journey today!
It consists of a network of capillaries surrounded by a specialized epithelial structure called the Bowman's capsule.
The glomerulus plays a crucial role in maintaining fluid balance, electrolyte homeostasis, and blood pressure regulation.
Accurate understanding of glomerular structure and function is essential for research on kidney disease, hypertension, and other related conditions.
PubCompare.ai's AI-driven platform can help optimize your kidney glomerulus research by identifying the best protocols and products through comprehensive comparisons of literature, preprints, and patents, enhancing reproducibility and accurracy with cutting-edg tools.
Start your research journey today!
Most cited protocols related to «Kidney Glomerulus»
Gene Expression
Homo sapiens
Kidney
Kidney Glomerulus
RNA
Tissues
Each participating center will enroll approximately 250 consecutive individuals over a 5-year period from 2011 until 2015, totaling 2,450 adult patients with CKD who provide written informed consent. The participating individuals will be monitored for approximately 10 years until death or until ESRD occurs.
The KNOW-CKD will enroll ethnic Korean patients with CKD who range in age between 20 years and 75 years. The CKD stages from 1 to 5 (predialysis), based on the eGFR, is calculated using the four-variable Modification of Diet in Renal Disease (MDRD) equation as follows:
eGFR (ml/min per 1.73 m2) = 175 × [serum Cr (mg/dl)] -1.154 × [age]-0.203 × [0.742 if female] × [1.212 if black], using serum creatinine concentrations measured at a central laboratory and an assay traceable to the international reference material [12 (link)].
Excluded subjects are those who 1) are unable or unwilling to give written consent, 2) have previously received chronic dialysis or organ transplantation, 3) have heart failure (NYHA class 3 or 4) or liver cirrhosis (Child-Pugh class 2 or 3), 4) have a past or current history of malignancy, 5) are currently pregnant, or 6) have a single kidney due to trauma or kidney donation.
We defined and allocated the specific causes of the CKD into four subgroups: glomerulonephritis (GN), diabetic nephropathy (DN), hypertensive nephropathy (HTN), and polycystic kidney disease (PKD). The definition of the subgroup is defined by the pathologic diagnosis, in the event that the biopsy result is available. Otherwise, the subgroup classification depends on the clinical diagnosis. GN is defined by the presence of glomerular hematuria or albuminuria with or without an underlying systemic disease causing glomerulonephritis. The diagnosis of DN is based on albuminuria in a subject with type 2 diabetes mellitus and the presence of diabetic retinopathy. HTN is defined by the patient’s hypertension history and the absence of a systemic illness associated with renal damage. Unified ultrasound criteria [13 (link)] will be used to diagnose PKD. The other causative diseases will be categorized as ‘unclassified’.
The KNOW-CKD will enroll ethnic Korean patients with CKD who range in age between 20 years and 75 years. The CKD stages from 1 to 5 (predialysis), based on the eGFR, is calculated using the four-variable Modification of Diet in Renal Disease (MDRD) equation as follows:
eGFR (ml/min per 1.73 m2) = 175 × [serum Cr (mg/dl)] -1.154 × [age]-0.203 × [0.742 if female] × [1.212 if black], using serum creatinine concentrations measured at a central laboratory and an assay traceable to the international reference material [12 (link)].
Excluded subjects are those who 1) are unable or unwilling to give written consent, 2) have previously received chronic dialysis or organ transplantation, 3) have heart failure (NYHA class 3 or 4) or liver cirrhosis (Child-Pugh class 2 or 3), 4) have a past or current history of malignancy, 5) are currently pregnant, or 6) have a single kidney due to trauma or kidney donation.
We defined and allocated the specific causes of the CKD into four subgroups: glomerulonephritis (GN), diabetic nephropathy (DN), hypertensive nephropathy (HTN), and polycystic kidney disease (PKD). The definition of the subgroup is defined by the pathologic diagnosis, in the event that the biopsy result is available. Otherwise, the subgroup classification depends on the clinical diagnosis. GN is defined by the presence of glomerular hematuria or albuminuria with or without an underlying systemic disease causing glomerulonephritis. The diagnosis of DN is based on albuminuria in a subject with type 2 diabetes mellitus and the presence of diabetic retinopathy. HTN is defined by the patient’s hypertension history and the absence of a systemic illness associated with renal damage. Unified ultrasound criteria [13 (link)] will be used to diagnose PKD. The other causative diseases will be categorized as ‘unclassified’.
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Adult
Biological Assay
Biopsy
Child
Creatinine
Diabetes Mellitus, Non-Insulin-Dependent
Diabetic Nephropathy
Diabetic Retinopathy
Diagnosis
Dialysis
Diet
EGFR protein, human
Glomerulonephritis
Heart Failure
Hematuria
High Blood Pressures
Hypertensive Nephropathy
Kidney
Kidney Diseases
Kidney Failure, Chronic
Kidney Glomerulus
Koreans
Liver Cirrhosis
Malignant Neoplasms
Organ Transplantation
Patients
Polycystic Kidney Diseases
Renal Agenesis, Unilateral
Serum
Ultrasonics
Woman
Wounds and Injuries
Brain
Gene Annotation
Gene Chips
Gene Expression
Genes
Homo sapiens
Hypertensive Nephropathy
IGA Glomerulonephritis
Kidney Glomerulus
Mice, Laboratory
Microarray Analysis
Mus
Poly A
RNA, Polyadenylated
Sample collection from Healthy population: For the determination of the reference interval, blood and urine samples were collected from healthy healthcare professionals (10 males and 10 females). Individuals were informed of the testing to be performed and only the specified testing was performed on the collected reference specimens. Analyses were conducted with the full respect for the individual's right to confidentiality. Information regarding their age, gender and race was also collected with the healthcare professional's consent to calculate the glomerular filtration rate (GFR). Urine creatinine and serum creatinine was assayed for each of these samples. Freshly collected urine samples were allowed to sit at room temperature for 30 minutes to sediment, and the supernatant was aliquoted and stored at -70 oC until analysis.
Construction of KIM-1 sandwich ELISA (R&D Cat# DY1750, Minneapolis, MN): The wells of Nunc-Maxisorp EIA plates were coated by diluting the capture antibody (72 μg/mL) to a working concentration of 0.4 μg/mL in PBS with 100 μL in each well. The plate was sealed and incubated overnight at room temperature. Each well was aspirated and washed using an automated microplate washer (Bio-Tek) with 400 μL of Wash Buffer (0.05% Tween-20 in PBS), repeating the process two times for a total of three washes. The plates were blocked by adding 300 μL of reagent diluent (1% BSA in PBS, 0.2 μm filtered) to each well and incubated at room temperature for 2 hours. After washing as in the previous step, 100 µL of standard recombinant human KIM-1 (0-2000 pg/mL), control, and urine sample was pipetted to the designated well, covered with an adhesive strip, and placed on the orbital shaker at 400 rpm at room temperature for 2 hours. The plate was washed using the same wash protocol as before, and 100 μL of the biotinylated goat anti-human KIM-1 detection antibody diluted in reagent diluent to a working concentration of 400 ng/mL was added to each well. The plate was covered with a new adhesive strip, incubated at room temperature for 2 hours with continued shaking at 400 rpm. Washing step was repeated and 100 μL of streptavidin-HRP diluted to a working dilution was added to each well. The plate was protected from light, covered with a new adhesive strip, shaken at 400 rpm, and incubated at room temperature for 20 minutes. After washing, 100 µL substrate solution was added to all wells, protected from light, covered with an adhesive strip, shaken at 400 rpm, and incubated at room temperature for 7 minutes. The reaction was stopped by adding 50 µL of stop solution to all wells. The absorbance was measured using a plate reader (BioTek Elx800) at 450 nm with an absorbance correction at 540 nm. The urinary KIM-1 concentration was calculated based on the standard curve and expressed in absolute terms (pg/mL).
Evaluation of the KIM-1 ELISA: The validation of the KIM-1 ELISA was evaluated by measuring linearity, intra-run precision, inter-run precision, analytical sensitivity, recovery, dilution verification, reference range, stability and length of run. Details of the criteria for each are described in the result section.
Sample collection and analysis: Urine samples were collected asceptically directly in urine cups and stored at -70 oC within 4 hours. All statistical analysis was carried out using the program EP evaluator 15 (version 8.0, DG Rhoads).
Analytical Testing: Urinary creatinine was measured by the Jaffe rate-blanked creatinine assay using a Roche/ Hitachi 911 system (Roche Diagnostics, Indianapolis , IN, USA). Serum enzymatic creatinine was measured using the Roche/Hitachi P800 analyzer. Glomerular filteration rate (GFR) was calculated using the MDRD (modification of diet in renal disease) formula 186 X (SerumCreatinine)-1.154 X (Age)-0.203 X (0.742 if female and 1.000 if male) X (1.210 if African American and 1.000 if others) (mL/min/1.73m2) 16 .
Construction of KIM-1 sandwich ELISA (R&D Cat# DY1750, Minneapolis, MN): The wells of Nunc-Maxisorp EIA plates were coated by diluting the capture antibody (72 μg/mL) to a working concentration of 0.4 μg/mL in PBS with 100 μL in each well. The plate was sealed and incubated overnight at room temperature. Each well was aspirated and washed using an automated microplate washer (Bio-Tek) with 400 μL of Wash Buffer (0.05% Tween-20 in PBS), repeating the process two times for a total of three washes. The plates were blocked by adding 300 μL of reagent diluent (1% BSA in PBS, 0.2 μm filtered) to each well and incubated at room temperature for 2 hours. After washing as in the previous step, 100 µL of standard recombinant human KIM-1 (0-2000 pg/mL), control, and urine sample was pipetted to the designated well, covered with an adhesive strip, and placed on the orbital shaker at 400 rpm at room temperature for 2 hours. The plate was washed using the same wash protocol as before, and 100 μL of the biotinylated goat anti-human KIM-1 detection antibody diluted in reagent diluent to a working concentration of 400 ng/mL was added to each well. The plate was covered with a new adhesive strip, incubated at room temperature for 2 hours with continued shaking at 400 rpm. Washing step was repeated and 100 μL of streptavidin-HRP diluted to a working dilution was added to each well. The plate was protected from light, covered with a new adhesive strip, shaken at 400 rpm, and incubated at room temperature for 20 minutes. After washing, 100 µL substrate solution was added to all wells, protected from light, covered with an adhesive strip, shaken at 400 rpm, and incubated at room temperature for 7 minutes. The reaction was stopped by adding 50 µL of stop solution to all wells. The absorbance was measured using a plate reader (BioTek Elx800) at 450 nm with an absorbance correction at 540 nm. The urinary KIM-1 concentration was calculated based on the standard curve and expressed in absolute terms (pg/mL).
Evaluation of the KIM-1 ELISA: The validation of the KIM-1 ELISA was evaluated by measuring linearity, intra-run precision, inter-run precision, analytical sensitivity, recovery, dilution verification, reference range, stability and length of run. Details of the criteria for each are described in the result section.
Sample collection and analysis: Urine samples were collected asceptically directly in urine cups and stored at -70 oC within 4 hours. All statistical analysis was carried out using the program EP evaluator 15 (version 8.0, DG Rhoads).
Analytical Testing: Urinary creatinine was measured by the Jaffe rate-blanked creatinine assay using a Roche/ Hitachi 911 system (Roche Diagnostics, Indianapolis , IN, USA). Serum enzymatic creatinine was measured using the Roche/Hitachi P800 analyzer. Glomerular filteration rate (GFR) was calculated using the MDRD (modification of diet in renal disease) formula 186 X (SerumCreatinine)-1.154 X (Age)-0.203 X (0.742 if female and 1.000 if male) X (1.210 if African American and 1.000 if others) (mL/min/1.73m2) 16 .
African American
Antibodies, Anti-Idiotypic
Biological Assay
BLOOD
Buffers
Creatinine
Diagnosis
Diet
Enzyme-Linked Immunosorbent Assay
Enzymes
Females
Glomerular Filtration Rate
Goat
HAVCR1 protein, human
Health Personnel
Homo sapiens
Hypersensitivity
Immunoglobulins
Kidney Diseases
Kidney Glomerulus
Light
Males
Population Health
Serum
Specimen Collection
Streptavidin
Technique, Dilution
Tween 20
Urine
Arteries
BLOOD
Cloning Vectors
Diet
Folate
Gene Delivery Systems
Institutional Animal Care and Use Committees
interleukin-1beta-converting enzyme inhibitor
Kidney
Kidney Glomerulus
Luciferases
Males
Mice, Inbred C57BL
Microbubbles
Mus
Obstetric Delivery
Physical Examination
Plasmids
Short Hairpin RNA
Tissues
Ultrasonography
Urine
Most recents protocols related to «Kidney Glomerulus»
Example 3
As shown in
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Biopsy
Cicatrix
Cytoplasm
Epithelial Cells
Epithelium
Kidney
Kidney Glomerulus
Nephrectomy
Patients
Podocytes
Proteins
Tissue Donors
Tissues
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American Indian or Alaska Native
Biopsy
Diabetes Mellitus
Digestive System
Ethics Committees, Research
Gene Chips
Gene Expression
Genome, Human
Genotype
Kidney
Kidney Diseases
Kidney Glomerulus
Living Donors
Microarray Analysis
Microdissection
Tissue Procurement
Tissues
Vulnerable Populations
Signals from a total of 14 bees were recorded and analysed. Data analysis was fully automated, based on custom MATLAB (R2019b, MathWorks) scripts. The fluorescence time series, containing an entire experimental sequence, were separated into periods of 3 s pre-stimulus, 3 s during the stimulus and 3 s post-stimulus for each trial. For each frame, we computed the relative fluorescence change via normalizing the raw fluorescence signal by the average signal during the pre-stimulus period Fb. This signal is proportional to the relative change in calcium concentration and thus the neuronal firing rate64 (link). ΔF/F was averaged over the 10 trials for each odour. Next, the 2D activity maps were segmented for individual glomerular responses. The glomerular boundaries were obtained by recursively comparing three sources of information, the anatomical features from an additionally recorded 3D image stack (Supplementary Fig. 2 a), the functional response maps (Supplementary Movie 2 ) and a regional homogeneity analysis that tests the correlation between the signals of each pixel and those from neighbouring pixels. This measure is high within individual glomeruli and falls off at their borders (Supplementary Fig. 2 b). After coherently responding structures were segmented, the glomerular identity was determined using the digital 3D antennal lobe atlas 23. The analysis was limited to the 19 most frequently identified glomeruli. If the identity of individual glomeruli could not be determined with certainty, they were discarded, so that the total number of analysed glomeruli fluctuated across bees (Fig. 4 c).
A statistical analysis of the subjects’ mean responses to single odours was performed via paired t-tests with FDR correction.
A statistical analysis of the subjects’ mean responses to single odours was performed via paired t-tests with FDR correction.
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Bees
Calcium
Fluorescence
Kidney Glomerulus
Microtubule-Associated Proteins
Neurons
Odors
Reading Frames
The monotonicity of the response x (x = maximal spike density function (SDF) or averaged SDF, each across all glomeruli) in this context was calculated as. i.e. the difference between the SDF at the highest tested odour concentration ( and the highest observed SDF, normalised by the mean observed SDF, where the maximum and mean were taken across all concentrations from to . Monotonicity takes values smaller or equal 0, and monotonic odours have .
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Kidney Glomerulus
Odors
The two-photon microscope (Ultima IV, Bruker) is based on an ultra-short pulsed laser (Mai Tai, Deep See HP, Spectra-Physics). The laser was tuned to 780 nm for fura2 excitation. All images were acquired with a water-immersion objective (10 , NA 0.3, Olympus). The fluorescence was collected in epi-configuration, selected by a dichroic mirror, and filtered with a band-pass filter centred at 525 nm and with a 70 nm bandwidth (Chroma Technology Corp). Finally, it was detected by a photomultiplier tube (Hamamatsu Photonics). Laser powers of about 10 mW were used in order to balance signal-to-noise ratio (SNR) against photo-damage effects that reduced the bee life span.
The field of view of 280 × 280 µm2 was resolved by 128 × 128 pixels. The fluorescence intensity was recorded with a depth of 13 bits. The image acquisition at a frame rate of 10.1 Hz was synchronized to the stimulus protocol.
In addition to the functional images, a z-stack of the antennal lobe was acquired with a spatial resolution of 512 × 512 pixels and a z-layer distance of 2 µm to perform the morphological identification of glomeruli.
The field of view of 280 × 280 µm2 was resolved by 128 × 128 pixels. The fluorescence intensity was recorded with a depth of 13 bits. The image acquisition at a frame rate of 10.1 Hz was synchronized to the stimulus protocol.
In addition to the functional images, a z-stack of the antennal lobe was acquired with a spatial resolution of 512 × 512 pixels and a z-layer distance of 2 µm to perform the morphological identification of glomeruli.
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Fluorescence
Kidney Glomerulus
Microscopy
Reading Frames
Strains
Submersion
Top products related to «Kidney Glomerulus»
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More about "Kidney Glomerulus"
The kidney glomerulus, also known as the renal glomerulus, is the fundamental filtration unit of the kidneys.
It is responsible for removing waste, toxins, and excess water from the bloodstream, playing a crucial role in maintaining fluid balance, electrolyte homeostasis, and blood pressure regulation.
The glomerulus consists of a network of capillaries surrounded by a specialized epithelial structure called the Bowman's capsule.
Accurate understanding of glomerular structure and function is essential for research on kidney disease, hypertension, and other related conditions.
Researchers can utilize tools like Image-Pro Plus 6.0, a powerful image analysis software, to study glomerular morphology and function.
Fetal bovine serum (FBS) is often used to culture glomerular cells, while Dynabeads, magnetic beads coated with specific antibodies, can be used to isolate glomerular cells for further analysis.
Techniques like TRIzol reagent and the RNeasy Mini Kit can be employed to extract and purify RNA from glomerular samples, enabling gene expression studies.
Antibiotics like penicillin and streptomycin are commonly used to prevent contamination in cell culture experiments involving the glomerulus.
The BX51 microscope, a high-quality optical microscope, can be utilized to visualize and analyze glomerular structures.
PubCompare.ai's AI-driven platform can help optimize kidney glomerulus research by identifying the best protocols and products through comprehensive comparisons of literature, preprints, and patents.
This can enhance reproducibility and accuracy, allowing researchers to make informed decisions and advance their studies on this crucial filtration unit of the kidneys.
Start your research journey today and discover how PubCompare.ai can support your efforts to understand the structure and function of the kidney glomerulus.
It is responsible for removing waste, toxins, and excess water from the bloodstream, playing a crucial role in maintaining fluid balance, electrolyte homeostasis, and blood pressure regulation.
The glomerulus consists of a network of capillaries surrounded by a specialized epithelial structure called the Bowman's capsule.
Accurate understanding of glomerular structure and function is essential for research on kidney disease, hypertension, and other related conditions.
Researchers can utilize tools like Image-Pro Plus 6.0, a powerful image analysis software, to study glomerular morphology and function.
Fetal bovine serum (FBS) is often used to culture glomerular cells, while Dynabeads, magnetic beads coated with specific antibodies, can be used to isolate glomerular cells for further analysis.
Techniques like TRIzol reagent and the RNeasy Mini Kit can be employed to extract and purify RNA from glomerular samples, enabling gene expression studies.
Antibiotics like penicillin and streptomycin are commonly used to prevent contamination in cell culture experiments involving the glomerulus.
The BX51 microscope, a high-quality optical microscope, can be utilized to visualize and analyze glomerular structures.
PubCompare.ai's AI-driven platform can help optimize kidney glomerulus research by identifying the best protocols and products through comprehensive comparisons of literature, preprints, and patents.
This can enhance reproducibility and accuracy, allowing researchers to make informed decisions and advance their studies on this crucial filtration unit of the kidneys.
Start your research journey today and discover how PubCompare.ai can support your efforts to understand the structure and function of the kidney glomerulus.