Renal - Science topic

EGTI is a reliable and informative assay to determine the degree of damage arising out of endothelial, glomerular, tubular and interstitial injury and hence utilized for reporting histological damage caused by kidney ischemia. However, the detailed method of calculation might be obtained by searching internet.

β-Caryophyllene: A Sesquiterpene with Countless Biological Properties

1📷by📷Fabrizio Francomano1,†,📷Anna Caruso1,†,📷Alexia Barbarossa1,📷Alessia Fazio1,*,📷Chiara La Torre1,📷Jessica Ceramella1,📷Rosanna Mallamaci2📷,📷Carmela Saturnino3,📷Domenico Iacopetta1📷 and📷Maria Stefania Sinicropi

1

Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, Via P. Bucci, 87036 Arcavacata di Rende, Italy

2

Department of Biosciences, Biotechnology and Biopharmaceutics, University of Bari, Via Orabona 4, 70124 Bari, Italy

3

Department of Science, University of Basilicata, 85100 Potenza, Italy

*

Author to whom correspondence should be addressed.

†

The authors have contributed equally to the manuscript.

Appl. Sci. 2019, 9(24), 5420; https://doi.org/10.3390/app9245420

Good evening,

It depends so much on each patient

If he is in pain, he is supposed to be admitted in the hospital in order to have i.v. medications like antispastics, antiinflamatory, pain medication, antibiotics, even opioids. When needed, lithotripsy.

It is necesary to determine the cause that induces kidney stones. It is important to establish if there is a local or systemic condition.

To prevent kidney stones: cure the cause, if possible, drink plenty of plain water, antispasmodic medication, when pain is present.

Some author encourage red bilberry extract.

The same hemodialysis' machine like fressenius or prismaflex get a funtion to measure hematocrit and sample hemodinamic curves

Also, see the link below.

Hi! Not sure if ImageJ has an equivalent plugin, but I find the 'Incremental Distance' tool on Image-Pro Premier very useful for measuring vessel wall thickness between two irregular shaped lines or polygons. Drawback though is that it's a paid software... a free version will definitely be useful to the general community. Good luck!

Video on its use on YouTube:

The incidence of this post-procedural complication is so rare that only one case is reported so far in Yemen.

Since the complication is rare, the management options recommended are just expert opinions.

The diagnosis of escaped stone is made intraoperatively and the best management option is to close the pyelolithotomy incision and proceed with intraperitoneal exploration to find the lost stone. In difficult circumstances, this may be impossible and can even be a reason for iatrogenic injury to intraperitoneal structures. In such difficulties, i think it is better to stop the procedure and return the patient to the ward. The incident can be discussed with the patient and further management options can be decided. Here, i think an open or laparoscopic peritoneal exploration is possible after localizing the stone with a imaging.

A surgeon should avoid open peritoneal exploration to remove the stone on the same surgery day since this might result in a legal allegation. Most patients choose laparoscopy for its cosmetic advantage and short hospital stay. Patient consent is mandatory if open exploration is decided and this is better be done few days or weeks after the first surgery.

I don't think an escaped stone can be managed conservatively. The evidences are scarce to comment on the place of conservative management. However, the decision to do so appears to be dependent on the patient's preference.

سؤال قيم كنت اتمنى الإجابة

That makes sense. Could you be referring to C0 instead of Cmax instead? C0 is the extrapolated value to time 0, which will be Dose/Vd. If C0 changed and your dosing is constant, it is likely that there was a difference in volume of distribution that occurred. I generally find Vd harder to explain biologically as the reason for the change could be anything from changed transporters in blood cells to fat or other peripheral tissues or plasma protein binding.

In IV data, Ke = CL/Vd. If CL changed, but Ke did not, it is possible that Vd also changed to similar extent. Ke = ln2/(t1/2), so if Ke does not change, t1/2 will not change. An easy way to see this would be (as a quick and dirty way of ensuring your calculations were correct), if you plot your plasma data on a log 10 y axis, do the gradients of each mouse group look similar? Ke is pretty much the slope on that graph. Here is a list of useful PK equations that I often reference to understand such plots better. https://pharmacy.ufl.edu/files/2013/01/5127-28-equations.pdf

It is also possible that the virus you were administering did not do what you expected. Perhaps other things to investigate post sac, e.g. kidney histology - did the virus damage the kidney tissue?

Ana Ortega Hello, can I ask you some tips for cultivating podocytes

Dr Cakiroglu,

I will be doing preferably in India

Thomas Monaghan how do I get into the twitter forum?

I've seen some papers in renal transplantation:

I hope this researchgate link will help you

SGLT2 inhibitors could have an impact in prevention/slowing down progression of nephropathy as it helps to lower the renal tubular threshold for glucose re-absorption, and glucosuria occurs at lower plasma glucose concentration

( Pharmacotherapy handbook).

Thank you for your ansewr But EG is a nephrotoxic molecule and can interfere with the used drug effect (medicinal plant extract ).

The middle sample is the first morning urine for proper testing. Usually, at least 3 morning urine samples on three different days in separate sterile containers recommended by the laboratory are required to verify the presence of TB germs. Because antibiotics inhibit the growth of TB germs in the urine, it is advisable that the patient does not take antibiotics before giving the sample. Samples less than 40 ml or held for more than 24 hours will not be suitable for culture. It is difficult to see TB germs in the urine sample and up to seven samples are recommended to increase the probability.

I Had no problem acessing the Petkova article

Yes, there are many studies where Immunohistochemistry staining done for Uromodulin using antibody against Uromodulin (Tamm-horsfall protein) as that done by Tarana Arman ,Washington State University

I have a question???

What is the reason that you can not find the article???????

Ingrid

Hi Ronald C. Van Gaal ,

Very interesting observations indeed. I just had a quick look at the tubulin antibody product sheet you are using and think that there is a good chance of the antibody binding being affected by tubulin PTMs. Alternatively, it cannot be ignored that the cells might be expressing different tubulin isotypes (there exists different tubulin isotypes, mostly divergent in the c-terminal part of the protein).

One way to understand this is to simply use tubulin antibody from another source. At the same time, you could also try to co-stain these cultures with tubulin PTM antibodies. Please see this reference for more details

"The tubulin code: Molecular components, readout mechanisms, and functions"

However, before concluding something, it will be better to co-stain these cultures with actin or other cellular markers (preferably apoptosis markers) and make sure that the cells showing negative staining for tubulin are alive and healthy.

Thanks

Satish

Empirically, we prefer to go for an USG of KUB to exclude any pathology (parenchyma) other than the specific scenario-based suspected causes of AKI along with other investigations. Although this's not conclusive. Regards- Rabiul.

Hello

sorry we do not have data base of renal stones

which is very important

thank you for your important and vital question

The combination elbasvir (a NS5A inhibitor) and grazoprevir (a NS3/4 protease inhibitor) is currently 1 of the 2 recommended regimens for patients with CKD stage 4 or 5. The recommended dosage is a daily fixed-dose combination of elbasvir (50 mg)/grazoprevir (100 mg) and treats patients with HCV genotype 1a, 1b, and 4 infections.

A landmark study highlights the potential of this regimen in patients on dialysis. The C-SURFER (Hepatitis C: Study to Understand Renal Failure’s Effect on Responses) study was a phase 3, randomized study involving 224 patients with CKD stage 4 or 5 with or without dialysis dependence and with HCV genotype 1 infection.23 Seventy-six percent of patients were dialysis-dependent, 81% had CKD stage 5 at baseline, and 80% were naive to treatment. Patients were randomly assigned to receive elbasvir 50 mg and grazoprevir 100 mg (immediate treatment group) or placebo (deferred treatment group) once daily for 12 weeks. The deferred treatment group was used as an internal control for potential safety signals in the immediate treatment group and started active treatment at week 16. The primary outcome was a SVR at 12 weeks, which was achieved by 94% of patients in the intention-to-treat analysis. In the modified full analysis (in which 6 patients were excluded due to reasons other than virologic failure), the SVR rate at 12 weeks was 99% . The most common adverse events were headache, nausea, and fatigue. Elevated lipase was the only drug-related adverse event reported. There was no consistent change in mean eGFR or creatinine.

Another retrospective study further demonstrated that the combination elbasvir/grazoprevir does not worsen kidney function in patients with preexisting renal disease and HCV infection.24 Patients with CKD stage 3 at baseline who were enrolled in elbasvir/grazoprevir phase 2/3 clinical trials with chronic HCV infection with or without ribavirin were included in the study (N=32). Thirty-one patients (97%) achieved SVR at 12 weeks. There was no decline in median eGFR at the end of treatment or at 12 weeks follow-up. In fact, many patients had an improvement in their CKD stage. All 32 patients had a glomerular filtration rate (GFR) of less than 60 mL/min at baseline; however, at the end of treatment, 12 patients had a GFR of at least 60 mL/min. Of note, this improvement was consistent regardless of the presence of cirrhosis, treatment with ribavirin, or HIV coinfection.

The combination glecaprevir (a NS3/4 protease inhibitor) and pibrentasvir (a NS5A inhibitor) has activity against all 6 major HCV genotypes with a recommended fixed daily dose of 300 mg and 120 mg, respectively. In the EXPEDITION-4 (Efficacy and Safety of ABT-493/ABT-530 in Renally Impaired Adults With Chronic Hepatitis C Virus Genotype 1-6 Infection) phase 3 trial, Gane and colleagues assessed the efficacy and safety of the combination regimen in patients with severe renal impairment (stage 4 or 5).25 One hundred and four patients who had chronic HCV genotypes 1 through 6 infections were enrolled and received the combination antiviral therapy for 12 weeks. Fifty-eight percent of patients had no prior treatment history, 81% had compensated cirrhosis, and 82% were on dialysis. The SVR rate at 12 weeks was 98% (102/104; Figure 1). The most common adverse events were pruritus, fatigue, and nausea, and the rates of adverse events between patients on dialysis vs patients not undergoing dialysis were similar (72% vs 68%, respectively). Four patients discontinued treatment due to adverse events; none of the serious adverse events were considered to be drug-related. Of note, 3 of the 4 patients who discontinued the treatment achieved SVR at 12 weeks. Pre- and postdialysis drug levels were similar for both drugs, with only a 3% to 6% difference.

The combination ledipasvir (90 mg) and sofosbuvir (400 mg) is approved for the treatment of HCV genotype 1 or 4 infection in kidney transplant patients. Colombo and colleagues performed a phase 2, open-label study in which 114 patients were given this regimen; 91% of patients had genotype 1 infection, 69% were treatment-naive, 15% had compensated cirrhosis, and the median GFR was 56 mL/min.32 Overall, 100% of patients achieved SVR at 12 weeks. Of note, only 4 patients had a decrease in GFR to less than 30 mL/min during therapy. In 3 of these patients, the GFR increased to more than 30 mL/min at the last visit recorded.

The combination glecaprevir (300 mg) and pibrentasvir (120 mg) is approved for the treatment of HCV genotypes 1 through 6 infections in kidney transplant patients. Reau and colleagues performed a phase 3, open-label, multicenter study in which patients without cirrhosis were treated for at least 3 months after either liver (80%) or kidney (20%) transplant.33 The median GFR was 62.3 mL/min. In total, 100 patients were enrolled and treated with glecaprevir/pibrentasvir for 12 weeks. Of the available data, 89 of 90 patients achieved SVR at 12 weeks; 1 patient experienced virologic failure. Another patient had mild liver transplant rejection but continued with treatment and achieved SVR at 12 weeks.

The combination sofosbuvir (400 mg) and daclatasvir (60 mg) plus a low initial dose of ribavirin (600 mg) has also been approved for kidney transplant patients. In the HCV-TARGET (Hepatitis C Therapeutic Registry and Research Network) study, 443 posttransplant recipients (60 kidney transplant patients, 347 liver transplant patients, and 36 dual liver-kidney transplant patients) were treated with several different regimens, including ledipasvir/sofosbuvir with or without ribavirin (85%), sofosbuvir/daclatasvir with or without ribavirin (9%), and ombitasvir/paritaprevir/ritonavir plus dasabuvir with or without ribavirin (Viekira Pak, AbbVie; 6%).34 Overall, 42% of patients had cirrhosis, and 54% were treatment-experienced, including 28 patients (6%) with prior exposure to the first-generation protease inhibitors telaprevir and boceprevir. The SVR rates at 12 weeks were 96% for liver transplant recipients, 94% for kidney transplant recipients, and 90% for dual transplant recipients. There were 6 episodes of acute rejection. Of note, ribavirin did not affect SVR rates.

HCV infection in patients with CKD is an important risk factor for morbidity and mortality. Advancements have been made in the treatment of this high-risk patient population with the approval of DAA agents in the treatment of chronic HCV infection. Elbasvir/grazoprevir and glecaprevir/pibrentasvir are the 2 recommended drug regimens for severe renal impairment and have SVR rates greater than 90%, which drastically change the opportunities for HCV eradication in this treatment population. For patients who are candidates for kidney transplantation, treatment after the procedure may be clinically reasonable; there are several regimens approved for such use. By increasing the number of patients treated for chronic HCV infection in both the dialysis and kidney transplant populations, a significant impact in global HCV eradication may be made

Have a look to the following link:

https://speciality.medicaldialogues.in/management-of-renal-and-ureteric-stones-nice-2019-guidelines/

I need the data for a presentation. It will be a great pleasure to collaborate in the future. Thank you.

As answered previously, radiopedia has specific cases such as this one: https://radiopaedia.org/cases/nephrolithiasis-and-post-pyelonephritic-scar

Paracetamol

Some drugs do not necessarily pass onto the milk but still have a negative effect on breastfeeding. Some drugs such as diuretics, testosterone, alcohol, oestrogens, progestins, ethanol etc may inhibit milk production and affect the success of breastfeeding. There are some that are classified as probably safe (check Hale's lactation risk categories), which are recommended to given only if benefit justifies the risk to the infant. For some of these, there is no evidence of any adverse effects from controlled studies. It makes sense to be cautious where safety is not known. It now calls for the practitioner to practice their own discretion and decide whether treatment can be deferred or an alternative drug can be administered.

Use suitable software. You ccan find on net

I wonder if this case can tolerate the surgery.?

Crystal growth is a complex issue. To sum it up. Having been formed a crystal, its growth depends on the solute concentration, its size and the spatial structure of the molecule. The factors that lead to crystals growth include factor such as: that the distance between crystal be small and the presence of Tamm-Horsfall glycoprotein (which acts as a glue).It is more frequently that in the kidney crystals grow in hyperosmolar sites as Henle loop and distal tubules. However, if its surface is healthy there are antiadhesive properties that inhibit this growth by means of the secretion of inhibitory macromolecules such as hyaluronic acid and osteopontin. Lastly, pH has a paramount influence in case of calcium fosfate stones, while calcium concentration is much more important in oxalate stones.

Hi,

Though I have never done these stains before, it appears the calcitonin-gene related peptide (CGRP) is a marker for afferent fibers in the renal area. You could double stain for Tyrosine Hydroxylase and CGRP and distinguish your two fiber types, similar to the approach taken in the attached paper. Those fibers not double positive for TH and CGRP can be assumed to be efferents.

Antibodies can be found here: https://www.biocompare.com/pfu/110447/soids/45900/Antibodies/CGRP

Clinics (Sao Paulo). 2016 Jan;71(1):22-7. doi: 10.6061/clinics/2016(01)05.

Six-minute walk test in children and adolescents with renal diseases: tolerance, reproducibility and comparison with healthy subjects.

Watanabe FT1, Koch VH2, Juliani RC1, Cunha MT1.

I think this case is a diffuse form of renal amyloidosis

Dear Mario,

I am sure that these cells will express HLA class I antigens (but not HLA-class II antigens) , too.

Best regards,

Thomas

Yes, in some of the literature about prescribing calcium it is noted that caution should be taken in administering this product to those with a history of stones.

As Emmanuel Obeagu points out, increased calcium intake will increase the chance of stone formation. However, this doesn't necessarily preclude use. There are a number of drug and disease interactions that should be considered though, especially in elders.

As for references, you might want to start with: Dietary Reference Intakes for Calcium and Vitamin D. Washington, DC: The National Academies Press; 2011 (https://www.ncbi.nlm.nih.gov/books/NBK56070/)

*Not a physician

Thank you very much for your comprehensive information and answers

.

Certain fatty acids and sphingoid bases found at mucosal surfaces are known to have antibacterial activity and are thought to play a more direct role in innate immunity against bacterial infections. Herein, we analysed the antibacterial activity of sphingolipids, including the sphingoid base sphingosine as well as short-chain C6 and long-chain C16-ceramides and azido-functionalized ceramide analogs against pathogenic Neisseriae. Determination of the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) demonstrated that short-chain ceramides and a ω-azido-functionalized C6-ceramide were active against Neisseria meningitidis and N. gonorrhoeae, whereas they were inactive against Escherichia coli and Staphylococcus aureus. Kinetic assays showed that killing of N. meningitidis occurred within 2 h with ω-azido-C6-ceramide at 1 X the MIC. Of note, at a bactericidal concentration, ω-azido-C6-ceramide had no significant toxic effect on host cells. Moreover, lipid uptake and localization was studied by flow cytometry and confocal laser scanning microscopy (CLSM) and revealed a rapid uptake by bacteria within 5 min. CLSM and super-resolution fluorescence imaging by direct stochastic optical reconstruction microscopy demonstrated homogeneous distribution of ceramide analogs in the bacterial membrane. Taken together, these data demonstrate the potent bactericidal activity of sphingosine and synthetic short-chain ceramide analogs against pathogenic Neisseriae.

V Manju, N Nalini. Effect of ginger on lipid peroxidation and antioxidant status in 1,2- dimethyl hydrazine induced experimental colon carcinog-enesis. J Biochem Tech (2010) 2(2):161-167 ISSN: 0974-2328

Z

It depends on application. For example in medical field we can not set cut off value to 0.05 but in social science we can set it.

Dear Singh Shivakumar,

good message. I agree with you that we should start treatment even stage 1 CKD, Because it is really as preclinical target organ damage. Also if Proteinuria with normal GFR is stage 1 CKD should be treated with first line ACEI or ARB. Main goal erly start treatment - to prevent progression of CKD. Thanks.

Other causes of hematuria:

1. Certain primary kidney diseases

2. Vigorous exercise or injury (falling off a bike and bruising a kidney)

3. Enlargement of the prostate (called benign prostatic hyperplasia), common problem in older men

4. Cancer of the bladder

5. Cancer of the prostate

6. Cancer of the kidney, (more often in patients over age 50 years)

7. Sometimes, the urine appears to have blood in it because there are other red substances (pigments) in the urine. This can happen if you eat an excessive amount of beets (called beeturia), food dyes, or certain medications (such as phenazopyridine/Pyridium)

Reference:

Dennis

Dennis Mazur

Dear Amal,

it will be different biomarkers.

As cardiologist I recommended for acute renal injury - NGAL.

Of course we have a lot of other biomarkers: tubular enzymes (α- and π-glutathione S-transferase, N-acetyl-glucosaminidase, alkaline phosphatase, γ-glutamyl transpeptidase, Ala-(Leu-Gly)-aminopeptidase, and fructose-1,6-biphosphatase), low-molecular weight urinary proteins (α1- and β2-microglobulin, retinol-binding protein, adenosine deaminase-binding protein, and cystatin C), Na+/H+ exchanger, neutrophil gelatinase-associated lipocalin, cysteine-rich protein 61, kidney injury molecule 1, urinary interleukins/adhesion molecules, and markers of glomerular filtration such as proatrial natriuretic peptide (1-98) and cystatin C.

For myocardial injury: hs Tn, Tn T ot I, MB-CK...

if you have 2 groups of animals and several variables measured in different time intervals, you need to run repeated measurement test for each. in spss general lenear model - repeated measures. nice explanation how to do and read and interprete the otcomes: https://www.youtube.com/watch?v=vhLS1yPax6M&t=121

katka

Dear Tatiyana,  I totally agreed with you. Many cases are in the literature where pacemaker and ICD implantation done for arrhythmias and latter on cardiac sarcoidosis was diagnosed on follow up. In our cases X-ray chest and cardiac CT was not suggestive Of cardiac sarcoidosis. Serum Angiotensin converting enzymes were also Normal. However cardiac MRI, PET scan or endocardial biopsy could have better investigation which was not done our cases. Over three years followup our cases showing no progesive left ventricular systolic dysfunction and patients are doing well. Wheather stress thallium scan could be of any use in our cases to look for microvascular dysfunction in 1st septal arterial territory? Wheather any small vessels cardiac vasculitis is a possibility? Sarcoidosis can involve heart by direct infiltration or by causing vasculitis of coronary arteries. Few case report are available where sarcoidosis was presenting with coronary vasculitis.. Thanks 

Thank you very much for your suggestion.

You are welcome

The etiology and pathophysiology of idiopathic recurrent acute pericarditis (IRAP) remain controversial and may involve both an infectious cause (usually viral or bacterial) as a trigger or an autoimmune and autoinflammatory cause in susceptible patients. IRAP is a rare disease of suspected immune-mediated pathogenesis. It represents a diagnosis of exclusion. At first it is necessary to rule out infectious and noninfectious causes of pericardial inflammation, including systemic autoimmune and immune-related disorders, because pericarditis may precede diagnosis of these disorders.  IRAP diagnosis is often made after a long follow-up. A minority of IRAP patients (6%) carry a mutation in the TNFRSF1A gene, encoding the receptor for tumor necrosis factor-alfa.

Thank you for your valuable suggestion.  This mathematical method can be applied for both both living and cadaveric donations.This method itself was developed on the basis of Prof. Oplez's findings. I shall definitely consult the results of CTS registry. 

Best Regards,

GS Muthu

HK-2 (human kidney 2) is a proximal tubular cell (PTC) line derived from normal kidney. The cells were immortalized by transduction with human papilloma virus 16 (HPV-16) E6/E7 genes. The cell line appears to be derived from a single cell based on Southern and FISH analysis.

There are 2 different types of Superficial Sideroses, one is Superficial Siderosis "of the central Nervous System", and the other is "Cortical" Superficial Siderosis, commonly seen in Cerebral Amyloid Angiopathy patients. If you're interested in the latter, you'll find ressources in the attached RG project ; Imaging biomarkers of hemorrhagic small vessel disease. Best regards 

Hi Sal,

PAE for symptomatic patients with BPH has been widely performed with low complication rates, however, there is a potential for severe complications, including technical and clinical treatment failures. Long catheter time after the procedure and repeated catheterization are seen frequently due to failed trials of voiding without catheter. We have experienced such worse outcome during conducting a comparative RCT between PAE and green light laser prostate ablation.

Furthermore, PAE has many limitations, including  lack of significant improvement in IPSS or Qmax in 25% of patients,  unknown long-term durability,  need for high dose ionizing radiation dose and contrast material for procedural guidance, can not be technically achieved on one or both sides as a result of  atherosclerosis, small artery size, and/or tortuosity and  collateral circulation may be present, and maintains vascularity.

To date, no high-quality multi-center RCTs have been published on safety, efficacy, and cost-effectiveness of PAE for BPH.  Most of the studies are of low quality due to their case series design. Therefore, advantages of PAE must be weighed against the risk of technical and clinical failures requiring a second intervention. 

Regarding your question, bladder neck will not be affected by a selective technique done by an expertise.

Regards

the rhythm start with low atrial tachycardia then switched to sinus tachycardia 

that is a common issue in worsening heart failure

I think correction of heart failure and its true precipitation would terminate this compensatory rhythm

The Bosniak classification groups cystic renal lesions into one of four categories based on CT/MR appearances. The differentiation between groups II and III is important as group II are typically ‘follow-up lesions’ and group III are ‘surgical lesions’. Features of a Bosniak III lesion include irregular thickened septa, measurable enhancement, coarse irregular calcification, multiloculation, nodularity, uniform wall thickening and margin irregularity. (Ref: Israel GM et al. Pitfalls in renal mass evaluation and how to avoid them. Radiographics 2008; 28: 1325–1338)

Actually with this new imformation, Prof. Marcel Machado gave a good answer.

In addition to normonatremic osmotic demyelination and PRES, CLIPPERS (Chronic lymphocytic inflammation with pontine perivascular enhancement responsive to steroids), may look similar on MRI but you would not expect spontaneous resolution.

Interesting perhaps, but as renal ptosis can cause hypertension, there is a differential diagnosis to consider, and it is not straight forward. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3628267/ Thus, one should check kidneys position and BP supine, and then repeat that erect, prior to exercise. Especially since ptosis repair is easy, with few complications.

Dear Konstantin,

Please check the following resources.

Best wishes

SM Najim

Flavio Sir- Will insulin be not required to keep rats alive for more than four weeks if using a dose of 35mg/kg of STZ along with high fat diet for inducing type 2 diabetes. By STZ administration blood glucose can increase to 400mg/dl and more so how to combat this.

I am sorry! I am a veterinary, so I do not have any patients. My research is in experimental surgeries in animal models, mainly in urogenital.

Thank you sir.

as far as i know therz no software of this sort...u vl have to do it manually under a microscope if u have H&E slides...

although this software thing is a good idea...needs to be developed...???

Hello,

The specific details of the answer to your first question depends upon what software you are using for the NLME analysis. It will revolve around setting up a "compartment" in the model which collects the drug eliminated renally.  This will then inform the estimation of the renal contribution to total clearance. Estimation of total clearance will be informed by the serum data.  It's a fairly simple process.

As for the second question, that's difficult.  If you can establish a good correlation between, say, amount of drug recovered in the urine during a certain time and AUC, then perhaps? Obviously renal clearance will need to correlate with total CL for this to work.

I will point out that just because a drug is "high renal extraction" does not mean it is mainly renally cleared.  It just means that the kidneys are very efficient at extracting the drug, but that doesn't mean the liver isn't the main clearance pathway.  The Fe indicates the importance of the kidneys to total clearance, not renal extraction ratio, or vice versa.  In your case the drug is mainly reanlly cleared though (Fe = 80%).

Regards,

David

Dear Ines

The concentrations are lower compared to brain tissue and the best detector for the job is an amperometric detector. Problem is that it requires isocratic elution profiles and the separation becomes a problem between these three analytes on C18 type columns. We have been trying to perform a wider range of biogenic amine detection in low volumes of plasma using LCMSMS and we can get down to pmole quantities after derivatisation. The problem is the close elution of compounds and the common fragment derived from the DANSYL group used to derivatize the biogenic amines results in cross-talk.

LCMSMS is a very powerful quantitative technique (that provides complementary information) with the choice instrument being a tripe quadrupole instrument.

PDA and ELSD are not sensitive enough for your application.

Good luck

Duncan

Nima, thanks for that protocol. It's very similar to the method I use for proximal tubules (which is vastly reassuring for me). The issue that I have is that I'm also looking to isolate collect duct cells for another experiment and was hoping to find a protocol that would allow me to isolate both. I've seen Percoll gradients being used but I'm really struggling to get it to work.

Thanks again for your help and i hope all is well.

Body surface area does not form urine. Even if it did, the BSA estimating formulas would not be BSA, but functions of weight to a power and height to a power. When some of us tested the statistical validity of using height in such a formula for correlation to a one exponential term model of 99m-Tc-DTPA clearance, we found that including height to a power and weight to a power decreased the correlation to GFR that would be achieved using a power of weight alone. That is, BSA is spuriously correlated to GFR (Look up "spurious correlation' as a search term, if you have not seen this before) . Much more accurate body scaling can be obtained using fractal geometry for which GFR is proportional to the 99m-Tc-DTPA volume of distribution to the 2/3 power and weight to the 1/4 power.

[See Wesolowski CA, Babyn PS, and Puetter RC. An improved method for determining renal sufficiency using volume of distribution and weight from bolus 99mTc-DTPA, two blood sample, paediatric data. Nuclear Medicine Communications 27(12): 961-968, 2006 and

Wesolowski CA, Babyn PS, and Puetter RC. A Power Law for Determining Renal Sufficiency Using Volume of Distribution and Weight from Bolus 99mTc-DTPA, Two Blood Sample, Pediatric Data. Nuclear Science Symposium Conference Record, 2006 IEEE, pp. 1986-1994, 2006.]

For those interested in body scaling, there are papers and books on the subject that have gone unnoticed by many. I might mention the work of West GB, Brown JH, Enquist BJ. The fourth dimension of life: fractal geometry and allometric scaling of organisms. Science 1999;284:1677-9, among many other papers. And the extensive review Agutter PS, Wheatley DN. Metabolic scaling: consensus or controversy? Theor Biol Med Model. 2004;1:13.

Now if one uses Kleiber's law namely GFR is proportional to weight to 3/4 power, better results are to be had (see publications above). However, GFR is not endogenous creatinine clearance. Because muscle mass increases as roughly the 1.08 power of weight, the creatinine clearance should use a lower power of weight for body scaling than GFR, and endogenous creatinine clearance is only roughly related to GFR, and, to make matters more confusing, the experimentally obtained power of weight for scaling creatinine is 0.69, and that for scaling inulin 0.77.

[See Adolph EF. Quantitative relations in the physiological constitutions of animals. Science. 1949;109:579-85.]

Finally, what do you want to use body scaling for? Errors in GFR measurement are proportional, Thus, the logarithm of GFR for a patient should be compared to a later logarithm of GFR of the same patient as logarithm of GFR errors are homoscedastic. However, taking logarithms does not normalize GFR (That is, convert GFR values into a normal distribution). For such, a different GFR transform is needed (information withheld, unpublished).

Normalization for drug dosimetry is an entirely different problem. For example, blood volume is approximately the first power of weight. That is, to calculate a dose of a vasoactive drug administered by intravenous infusion (example dipyridamole) one uses weight because independent of the body size of mammals, blood volume is roughly 5% of weight. This contrasts to dosing furosemide that I would dose by metabolism, that is, using for example

Pediatric dose = Adult dose * (W/70)^0.75

Next problem, how does one measure GFR? The best method is not affected by alterations in body fluid and would require few samples drawn over the shortest time possible. That is, as far as some of us can tell, the Tk-GV method

[Wesolowski CA, Puetter RC, Ling L, Babyn PS. Tikhonov adaptively regularized gamma variate fitting to assess plasma clearance of inert renal markers. J Pharmacokinet Pharmacodyn. 2010;37:435-74.

Wesolowski CA, Ling L, Xirouchakis E, Burniston MT, Puetter RC, Babyn PS, Giamalis IG, Burroughs AK. Validation of Tikhonov adaptively regularized gamma variate fitting with 24-h plasma clearance in cirrhotic patients with ascites. Eur J Nucl Med Mol Imaging. 2011;38:2247-56. doi:10.1007/s00259-011-1887-9.

Wickham, Fred; Burniston, Maria Teresa; Xirouchakis, Elias; Theocharidou, Eleni; Wesolowski, Carl Adam; Hilson, Andrew J W; Burroughs, Andrew Kenneth. Development of a modified sampling and calculation method for isotope plasma clearance assessment of glomerular filtration rate in patients with cirrhosis and ascites. Nucl Med Commun 34:1124–1132, doi:10.1097/MNM.0b013e32836529ab, 2013

And shortly, Surajith N. Wanasundara, Michal J. Wesolowski1, Mark C. Barnfield, Michael L. Waller, Anthony W. Murray, Maria T. Burniston, Paul S. Babyn, Carl A. Wesolowski1, Accurate and precise plasma clearance measurement using four Tc-99m-DTPA plasma samples over four hours, In press in Nuclear Medicine Communications, Sept. 9, 2015]

In summary, BSA is appropriate for estimating percentage damage of body surface area for burn victims as a guide to fluid replacement, but, there are few other situations for which I would not use a better estimator.

Goldblatt Hypertension Procedure (2K1C):

 Pre-surgical preparation:

 Fasting not required.

 • Inject analgesic (Buprenorphine at 0.02mg/Kg (SC), Written for Domitor anesthesia, but that will depend on your circumstances and your veterinarian. Domitor at 0.35mg/Kg (SC)) and antibiotic (Tribrissen at 0.25mL/Kg (SC)) 10 min prior to surgery.

Upon injection of domitor, dim the lights, cover the cage with a towel and remain silent for 5 min to allow sedation of rat (domitor renders animal hypersensitive to stimuli).

• Once the rat is asleep, shave the right flank and disinfect with 70% EtOH, 4% (w/v) hibitane and 10% iodine in a designed prepping area.

• Remove excess with warm, sterile water.

Surgery:

 • Transfer the rat onto the surgical table and set anesthetic to 1% isoflurane (0.5-0.6 L/min oxygen).

•  Adjust Bain circuit so as to see breathing of the rat in air reservoir bag.

• Cover rat with a sterile surgical drape.

• Make a 3-4 cm lateral incision in the skin (we use right flank, but left renal artery is longer and more accessible), ~ 0.5 cm from the ribs.

• Lift and cut through the muscle layer. Be careful not to damage any internal organs.

• Keep incision open with a retractor.

• Expose kidney and hold in place with custom-made retractor.

• Under dissecting microscope, expose a portion of the renal artery (~0.5 cm, half way between aorta and kidney) and clip it (silver neuro clip, Fine Science Tools).

- Diameter of renal artery distal to clip should be ~30-50% smaller in size after clipping & distal flow pulse should be apparent.

• Close muscle layer with interrupted sutures (4-0 vicryl).

• Close skin layer with hidden uninterrupted sutures.

• Inject Antisedan (1/4  dose for implant, 1/8 dose for 2K1C).

 Monitor vitals during surgery.

 - Relevant web site: http://www.transonic.com/rats1.shtml

Post-surgical monitoring:

Keep rat warm until he awakes (single-caged).

Buprenorphine given every 8-12 hours (3-4 injections or as required).

1. Critical period after surgery: 24-48 hours post-op

On surgery day monitor every hour for the first few hours.

Monitor 3 times/day for the next few days.

- Immediate signs of trouble:  chewing, incessant licking, pulling on sutures.

- What to do:  keep them from reaching the incision site (bandage or   anything that works)

Give post-op toys (cardboard) for the first few days to keep the rats distracted. Replace as they destroy them.

Give pipe toys only for the rest of the experiment.

2. Signs of potential pain and distress. Monitor closely and inform animal care staff.

·      Appearance:                      poor grooming

                                                      lacrimal discharge

                                                      piloerection

                                                      postural change (limping, hunching)

·      Behaviour:                         lack of grooming and nesting

                                                      lethargy

                                                      unresponsive to stimuli

                                                      abnormal vocalization

                                                      self-mutilation

·      Physical condition:           dehydration

                                                      weight loss

·  Vital signs:               temperature

heart rate

                                                      respiratory rate

                                                      blood pressure

Sacrifice rat if weight loss > 10%/day (at any time).

yes i agree particularly in stones

My patient was transplanted just 6 weeks ago, and has done very well. After her MAS she was treated with pulse solumedrol x 3 doses, followed by oral steroids 60mg q day and MMF in a dose of about 1000mg/m sq /day. She became remarkably well after going on the this treatment. She had been ill on hemodialysis for months without any diagnosable cause.  Oral Steroids were weaned completely as she was doing so well.

For the renal transplantation she was induced with Simulect and treated with Tacrolimus and MMF (our usual protocol), and she has been doing well except delayed graft function for 2 weeks. She is off dialysis now with almost normal renal function. Her MAS was not reactived during any of the surgeries.

Thanks for comments for my question

Most current methods of measuring GFR for a group of patients produce a long right tail of values corresponding to an inability of determining proper values for larger individuals. This can be mitigated by doing proper body scaling, and by obtaining GFR values using methods that have no such bias.

Body scaling in humans is most often incorrectly assumed to be related to body surface area, which thinking does not pass even the most cursory of thought experiments, never mind scientific investigation. For example, it should be obvious that skin surface area does not form urine; there is no causal relationship. However, since Sarrus F and Rameaux J. presented that silly idea in Paris on a hot summers day, 23 July 1839 to be exact, it has been repeated mindlessly. As Sarrus stood presenting his silly idea that a body radiates heat in proportion to his skin surface area, in my minds eye I picture him sweating in the sweltering heat, with his own perspiration showing him to be uttering nonsense. Humans sweat and primarily lose heat in proportion to their need to do so.

Much to my frustration, once a silly idea takes hold it is almost impossible to displace. In 2006, we tested this body surface concept and found it to be spurious. Wesolowski CA, Babyn PS, Puetter RC. An improved method for determining renal sufficiency using volume of distribution and weight from bolus Tc-99m-DTPA, two blood sample, paediatric data. Nucl Med Commun. DEC 2006 2006;27(12):963-970.

At the same time we showed how to better determine expected renal function.

While on the topic of silly ideas related to an inability to test kidney function in the obese, the next one is that elimination in the body follows a first order kinetic, which assumes that mixing of an injected substance in the body is instantaneous. Once again, this does not pass an even cursory thought experiment. It should be obvious that if someone injects an exogenous marker into the body, it will not be found in high concentration in the eyeball for many hours. A more appropriate kinetic is that an injection dilutes more gradually in time as time elapses. Wanasundara SN, Wesolowski MJ, Puetter RC, et al. The early plasma concentration of 51Cr-EDTA in patients with cirrhosis and ascites: a comparison of three models. Nucl Med Commun. 2015;36:392–397.

Obviously, for a larger individual, mixing takes longer, and the assumption of instant mixing leads to worse estimates of kidney function. So, what is needed is a model that is unaffected by fluid excess. Wesolowski CA, Ling L, Xirouchakis E, et al. Validation of Tikhonov adaptively regularized gamma variate fitting with 24-h plasma clearance in cirrhotic patients with ascites. Eur J Nucl Med Mol Imaging. 2011;38(12):2247-2256.

In summary, if one measures GFR more correctly, and if one knows what the expectations are for an obese person's kidney function, then one can expect to determine if that person has or does not have kidney disease. With the methods in current use, one cannot determine that with any accuracy. However, if the obese person is already in renal failure, or has an ongoing renal pathology that is already clinically florid, then yes, one can observe that with blood chemistry markers. And so it shall remain until better methods are taken seriously.

Tough question... markers do change from original isolation and during process and more during culture... CD133 is positive in other progenitors as well...  I would go for the functional part of the work.... as there are 'pulmospheres' and 'neurospheres'... have analog structures been described for renal tissue?  if not, grow and 'invent' them.... at least it will be a way of generating novel data that no one could argue against..... good luck and let us know.... defining progenitors  is a black whole on the stemmy way....

Hi, Elton, so far, we cannot retrieve the specific articles you want perhaps nobody has yet published one. However, the below 5 related articles, I hope, you may find them useful.

There are to my knowledge no studies on the possible value of such monitoring. In general, monitoring urinary output is of great value and add to fluid balance management. It may aslo signal disease onset or perturbed vascular homeostasis. However detailed calculation of in- vs output is often flawed for several reasons so monitoring urinary output mainly serves the role of monitoring...-urinary output...!

In the NICU urinary output is most commonly evaluated q 24, 12, 8 or (not so commonly) 4 h

Likely organisms (in U.S.) based on review by Gadkowski and Stout "Cavitary Pulmonary Disease" in Clin. Microbiol. Rev. April 2008 vol. 21 no. 2 305-333, full text at: http://cmr.asm.org/content/21/2/305.full

"The spectrum of illnesses associated with cavitary lung lesions is also broad among persons with immunosuppression due to malignancy or transplantation, but some specific illnesses are considerably more common in this population than in persons infected with human immunodeficiency virus. Patients who have received lung transplants frequently have posttransplant pneumonia caused by Pseudomonas, Staphylococcus, mycobacteria, and Aspergillus species, any of which may cause cavities. These infections are particularly common among persons with cystic fibrosis who received a transplant…Aspergillus should always be strongly suspected in the setting of cavitary lesions after transplantation… Invasive aspergillosis frequently occurs in the early posttransplant period, but with current prophylactic antifungal protocols, invasive aspergillosis is increasingly diagnosed in the later posttransplant period. P. jiroveci and Nocardia infections are also relatively common causes of lung infection among patients with hematological malignancy or posttransplantation, but the routine use of trimethoprim-sulfamethoxazole for Pneumocystis prophylaxis and the prevention of cytomegalovirus disease (a disease that increases the risk for other infectious complications) have significantly reduced the frequency of Pneumocystis and Nocardia lung infection in these patient populations. Of course, many of the pathogens discussed in this review can present as cavitary lesions in patients with hematological malignancy or transplants, so aggressive microbiological sampling, including tissue specimens, where possible, and appropriate testing (e.g., culturing for fungi and mycobacteria) are essential."

First it is important to differentiate Crohn's from cellcept/mycophenolate GI toxicity. They look very similar on colonoscopy or biopsy. If Crohn's is confirmed, treatment is usually sequential, involving antibiotics and then higher dose steroids.  Refractory cases may require anti-TNFa therapy - very effective but higher risk of infections long-term in combination with anti-rejection medications.

Dear Alireza,

I don't think I would agree with Dr.Theodor Klots, with regards to the timing of RAE.

One week is too early-ideally it is done just a day before surgery-sometimes even hours before.You want to operate before tumor lysis syndrome kicks in. Also, the longer one waits, the more edema and destruction of surgical planes is there, especially at the hilum.

In fact, there are a few papers, w