Nicole Gillain-Martin

Immunology, Human Biology

23.81

Questions and Answers

  • Nicole Gillain-Martin added an answer in Iron:
    4
    Is there prognostic or predictive significance when iron profile testing reveals an exaggerated response to IV iron?

    Is there prognostic or predictive significance in those approximately 5% dialysis patients who experience repeat exaggerated responses to even a single administration of modest doses of intravenous iron? [much higher than expected and more prolonged elevations of TSAT (serum iron/TIBC) and ferritin]?

    Sometimes, the labs remain elevated for months after a modest IV iron administration.

    While these labs are not perfect, they are currently the most commonly used markers to aid prescribing of iron therapy in dialysis.

    This observation presents challenges for the treating prescriber.

    It would be nice to know what is the significance to help guide therapy.

    I'm not familiar enough with the use of soluble ferric pyrophosphate (SFP) to know if SFP would be a better choice for treatment of these patients.

    I do not have full access to the below referenced article. While the focus may be different from the question above, I wonder if the authors noticed any significance for the observations referenced above.

    Nephrology (Carlton). 2016 Jan 29.
    Characterisation of hepatic and cardiac iron deposition during standard treatment of anaemia in haemodialysis.
    Holman R1, Olynyk JK1,2, Kulkarni H3, Ferrari P4,5.

    Nicole Gillain-Martin

    Do these patients actually had iron deficiency? Do you know this article?

  • Nicole Gillain-Martin added an answer in Renal Failure:
    5
    Is anyone familiar with the proteins used as markers for renal failure?

    I want to know the specific proteins for determination of renal failure.

    Nicole Gillain-Martin

    Serum Cystatin C and beta2-microglobulin are well known markers to highlight a renal impairment. What do you call renal failure? when renal failure is present at an advanced stage, and  when patient must be dialyzed soon, you will observe a high concentration of cystatin C in urine.

  • Nicole Gillain-Martin added an answer in Preeclampsia:
    4
    Can someone advise on the use of markers for preeclampsia such as PLGF and sFlt? How useful are these markers clinically?

    My question is why and how to implement the

    biomarkers sFlt-1 (soluble FMS-like tyrosine kinase) and PlGF (Placental growth

    factor) into clinical practice?

    Nicole Gillain-Martin

    You will find a lot of information by consulting this email address

  • Nicole Gillain-Martin added an answer in Clinical Studies:
    1
    Why the 24 hours urine is used in clinical studies and what is the duration of these?

    there are many protocols using the urine of 24 hours to study the phenomenon of cristatlisation

    Nicole Gillain-Martin

    There is no better method for estimating all the fluctuations of the composition of the urine depending on the amount of liquid ingested, the type of food eaten, the diurnal variations, the influence of the lying/standing position...

    It is essential that urine are properly collected: the patient must be well informed of the procedure

    The laboratory also plays an essential  role in the quality of the results:
    it is necessary that the entirety of the collected urine are mixed in a suitable sized container and that urine output is measured accurately (for that, use graduated cylinders of different sizes, not just use approximate measurements from some containers)

  • Nicole Gillain-Martin added an answer in Cystatin C:
    6
    Is creatinine or cystatin C a more accurate test for GFR and renal function in HIV positive patients?

    In studies monitoring tenofovir-associated kidney impairment either in HIV-positive patients on ART or negative patients on PrEP, a common test is serum creatinine levels, which are used to estimate glomerular filtration rate (GFR). However, some studies have shown that an alternative, cystatin C estimated GFR is more closely associated chronic kidneys disease outcomes. What is the state of the science for most accurately monitoring kidney function in patients on ART or PrEP? Thanks.

    Nicole Gillain-Martin

    I believe that the problem of patients who have AIDS is the same as that of other patients: marker and the formula used performances depend on the patient's weight, muscle mass and the value of the glomerular filtration rate.(The age also of course but I do not think there is currently elderly patients with AIDS). The CKD-EPI combined cystatin / creatinine formula looks best in various populations. Articles sent by A. Gagneux-Brunon also confirms this for these patients.

  • Nicole Gillain-Martin added an answer in Jaundice:
    16
    what is the possible type for cholestatic jaundice in a male neonate 21 days old with direct bilirubin of 2.4mg/dl?

    A male neonate 21 days old who is breast fed, with positive moro reflex, with good weight gain and yellow colored stool.

    On day 9: TSB 9.2mg/dl, PCV =50

    On day 11: TSB 8.6mg/dl; Direct bilirubin 2.5mg/dl; PCV 50

    On day 14: TSB 8.2; Direct bilirubin 2.2mg/dl; PCV 50

    On day 21: TSB 6.5mg/dl; direct bilirubin 2.5mg/dl; PCV 43

    Non fasting Ultrasound: normal size and texture for kidneys, spleen, liver, gallbladder and common bile duct, with no stones

    Day 22: Stop breastfeeding and start formula milk for 48 hours

    Day 25: TSB 7.6mg/dl; Direct bilirubin 2.4mg/dl; PCV 41

    So what is the expected diagnosis for this case? 

    Nicole Gillain-Martin

    The child has colored stools so I guess it has no sign of biliary atresia. Check if there is not a deficit alpha-1-antitrypsine.

  • Nicole Gillain-Martin added an answer in Adolescents:
    7
    What are the available searches for renal damage in obese adolescents?

    In earlier research we realized a high index of obese adolescentes. At the moment we are considering the possibility of kidney damage in these patients

    Nicole Gillain-Martin

    I agree with Mr. Wesolowski, there is no simple method (serum creatinine determination and application of a formula) to determine GFR in obese adolescents and adults. You have to use a method such as iohexol (easy to dose) clearance or why not determine the creatinine clearance through 24H urine. This method can highlight hyperfiltration.

  • Nicole Gillain-Martin added an answer in IgG:
    4
    A friend wanna be pregnant but her analysis showed: Toxoplasmosis IgG 188 and cytomegalovirus IgG 290. Is she in trouble?

    She is free of symptoms.

    Nicole Gillain-Martin

    If IgM are negative for both tests, they are old infections. The problem arises when IgM are positive because they can persist for long periods and do not necessarily reflect recent infection. A control after three weeks revealing a stabilization of antibodies is reassuring: infections are not new.
    One can also determine the IgG avidity: this is high when the infection is old

  • Nicole Gillain-Martin added an answer in IgG:
    4
    A friend wanna be pregnant but her analysis showed: Toxoplasmosis IgG 188 and cytomegalovirus IgG 290. Is she in trouble?

    She is free of symptoms.

    Nicole Gillain-Martin

    How long is she pregnant? As she made tests before pregnancy? IgM has it been realized? Without previous analysis, we can conclude nothing but it is likely older infections

  • Nicole Gillain-Martin added an answer in Albuminuria:
    3
    Can i determine eGFR without determining albuminuria?

    i mean eGFR can be determined by 3 variables: age , serum creatinine and sex according to formula it will be calculated.

    Nicole Gillain-Martin

    Albuminuria has nothing to do with the calculation of GFR, it is simply an additional criteria of severity of kidney disease. In contrast, albuminemia  is used in some MDRD formulas. I advise against using it. The latest international criteria advocate CKD-EPI formula based on creatinine.

  • Nicole Gillain-Martin added an answer in Bradford Assay:
    6
    Witch wavelength is best for measuring protein in the Biuret test?

    Witch wavelength is best for measuring protein in the Biuret test? I came across different variants ranging from 540-565nm, with the reagent consisting of the same chemicals. I tried measuring at all of the wavelengths but there are big differences, really significant for my results, and I wonder which is the most appropriate.

    Also the Biuret test gives me like 2-times more protein than I detect with the Bradford assay. This cannot be due to smaller peptides, I am pretty sure I don’t have any degraded protein in the sample. Any clue why I get this?

    Nicole Gillain-Martin

    The biuret method is based on detection of peptide bonds and is used to assay the proteins in a liquid which contains many proteins (> 5 g / l). The absorption spectrum of the complex formed is wide and the choice of the wavelength is not very important. The Bradford assay uses coomassie blue that reacts in an alkaline medium with different amino acids. It is generally used to assay small amounts of proteins. If I understand, you have standardized with an albumin solution. The proteins that are present in your solution may be very different from that protein. The intensity of the coloration of the two techniques can be influenced by the amino acid composition of your proteins. Think also to verify that the Beer-Lambert law is respected with both techniques in the area of concentration of your protein mixture

  • Nicole Gillain-Martin added an answer in Protein Staining:
    3
    How do I perform densitometric analysis of ponceau s. stain?

    Hello all, 

    I'm very new to research, and science in general. I just started in my first lab two months ago, so I don't have a lot of experience. 

    I'm currently analyzing western blot data. My PI and I are having some disagreement as to the best method to normalize the protein of interest (Neuropsin) to a total protein stain using Ponceau S.

    When doing densitometric analysis with AlphaView software of the Ponceau stain, I quantify the total protein for each sample using the entire lane (meaning I make the box to be analyzed around the whole lane). My PI tells me that I should be using only a small band from each lane instead of the whole lane. But it seems to me that this defeats the purpose of quantifying the total protein for each sample because it only quantifies a portion of the total protein. If I'm analyzing the entire band of my protein of interest, but only a portion of the total protein, it seems that the data from normalization won't be reliable. 

    Not much literature actually reports densitometric techniques, but in the papers that do, it appears that most researchers are using the entire lane to quantify total protein. Could anyone help clear this up for me? I want to make sure I'm using the most reliable methodologies.

    Thank you all ahead of time, I appreciate any responses. 

    Nicole Gillain-Martin

    I guess the soft from your densitometer allows you to quantify the importance of your protein relative to all proteins. If you have previously determined the total proteins concentration of your sample, you can calculate the concentration. Maybe,  the densitometer can estimate the concentration of a band on the intensity of staining the Ponceau because it has been previously calibrated to do so. In this case, only the reading of the band is sufficient. Both methods are valid only if the concentration of the bands does not exceed a certain value because the beer-lambert law must be respected

  • Nicole Gillain-Martin added an answer in Troponin:
    6
    Do you think measuring troponine is a useful biomarker for evaluation of Bupvacaine cardiotoxicity?

    Hi, I want to evaluate a drug on reducing the cardiotoxicity of Bupivacaine in dogs.  I searched the related articles but they haven't measured this biomarker in their assements !! What are your suggestions?

    Nicole Gillain-Martin

    The test may be tried, but then used a technique called "ultra-sensitive". Ensure also that the antibody used in the kit recognizes well the dog troponine

  • Nicole Gillain-Martin added an answer in Loop of Henle:
    9
    How does “Randall’s plaque” promote renal stone formation?

    Eight decades ago, Al exander Randall identified calcium phosphate deposits at the tip of renal papillae as the origin of renal calculi. The awareness that these “Randall’s plaque” promote renal stone formation has been amplified during the past years by the development of endoscopic procedures allowing the in situ visualization of these plaques. Recent studies based upon kidney biopsies evidenced that apatite deposits at the origin of these plaque originate from the basement membranes of thin loops of Henle and then spread in the surrounding interstitium. In addition, scanning electron microscopy examination of calcium oxalate stones developed on Randall’s plaque evidenced that plaque may also be made of tubules obstructed by calcium phosphate plugs. Hypercalciuria has been associated to Randall’s plaque formation. However, several additional mechanisms may be involved resulting in increased tissular calcium phosphate supersaturation and the role of macromolecules in plaque formation remains elusive. At last, apatite crystals are the main mineral phase identified in plaques, but other calcium phosphates and various chemical species such as purines have been evidenced, revealing thereby that several mechanisms may be responsible for plaque formation.

    Urolithiasis August 2014 Date: 07 Aug 2014
    Randall’s plaque as the origin of calcium oxalate kidney stones
    Michel Daudon, Dominique Bazin, Emmanuel Letavernier

    Nicole Gillain-Martin

    For Urolab India

    What do you suggest as a simple procedure to consider a supersaturation? In practice, using my knowledge, one use  the concentration of Ca, Mg, uric acid, oxalic acid and citric acid in urine.

  • Nicole Gillain-Martin added an answer in Loop of Henle:
    9
    How does “Randall’s plaque” promote renal stone formation?

    Eight decades ago, Al exander Randall identified calcium phosphate deposits at the tip of renal papillae as the origin of renal calculi. The awareness that these “Randall’s plaque” promote renal stone formation has been amplified during the past years by the development of endoscopic procedures allowing the in situ visualization of these plaques. Recent studies based upon kidney biopsies evidenced that apatite deposits at the origin of these plaque originate from the basement membranes of thin loops of Henle and then spread in the surrounding interstitium. In addition, scanning electron microscopy examination of calcium oxalate stones developed on Randall’s plaque evidenced that plaque may also be made of tubules obstructed by calcium phosphate plugs. Hypercalciuria has been associated to Randall’s plaque formation. However, several additional mechanisms may be involved resulting in increased tissular calcium phosphate supersaturation and the role of macromolecules in plaque formation remains elusive. At last, apatite crystals are the main mineral phase identified in plaques, but other calcium phosphates and various chemical species such as purines have been evidenced, revealing thereby that several mechanisms may be responsible for plaque formation.

    Urolithiasis August 2014 Date: 07 Aug 2014
    Randall’s plaque as the origin of calcium oxalate kidney stones
    Michel Daudon, Dominique Bazin, Emmanuel Letavernier

    Nicole Gillain-Martin

    For anyone interested, the publication of Prof. Bazin over Randall's plaque formation is available in its entirety on ResearchGate in its publications list. Thank you.

  • Nicole Gillain-Martin added an answer in Loop of Henle:
    9
    How does “Randall’s plaque” promote renal stone formation?

    Eight decades ago, Al exander Randall identified calcium phosphate deposits at the tip of renal papillae as the origin of renal calculi. The awareness that these “Randall’s plaque” promote renal stone formation has been amplified during the past years by the development of endoscopic procedures allowing the in situ visualization of these plaques. Recent studies based upon kidney biopsies evidenced that apatite deposits at the origin of these plaque originate from the basement membranes of thin loops of Henle and then spread in the surrounding interstitium. In addition, scanning electron microscopy examination of calcium oxalate stones developed on Randall’s plaque evidenced that plaque may also be made of tubules obstructed by calcium phosphate plugs. Hypercalciuria has been associated to Randall’s plaque formation. However, several additional mechanisms may be involved resulting in increased tissular calcium phosphate supersaturation and the role of macromolecules in plaque formation remains elusive. At last, apatite crystals are the main mineral phase identified in plaques, but other calcium phosphates and various chemical species such as purines have been evidenced, revealing thereby that several mechanisms may be responsible for plaque formation.

    Urolithiasis August 2014 Date: 07 Aug 2014
    Randall’s plaque as the origin of calcium oxalate kidney stones
    Michel Daudon, Dominique Bazin, Emmanuel Letavernier

    Nicole Gillain-Martin

    It is impossible to read your interesting publication in its entirety. Could you send it to us in a different way?
    (I think this is the problem announced by Mr Khan)

  • Nicole Gillain-Martin added an answer in Creatinine:
    5
    What is the best method for measuring urinary protein and creatinine concentration in mouse samples?

    Looking for a reliable testing kit for basic science research

    Nicole Gillain-Martin

    I agree with Mr Sanchez-Jacinto, the enzymatic method is more accurate than the Jaffe method, but in your case, as you work on mouse urine you have less interference of bilirubin, ketone bodies, médications. The Jaffe method is much more economical.
    The pyrogallol red method is quite satisfactory. 

  • Nicole Gillain-Martin added an answer in Chronic Renal Failure:
    6
    What importance do you attribute to the type of protein responsible for proteinuria?

    The intensity of proteinuria is an important element to define the degree of chronic renal failure. As a clinician, what importance do you attribute to the type of protein (low/high molecular weight) detected?

    Nicole Gillain-Martin

    A big thank you for all the answers that confirm the importance I have always attributed to have reliable techniques to quantify or separate these proteins by electrophoresis. This opinion is not shared by all nephrologists some being especially attached to the quantitative aspect of proteinuria

    About RBP: a study was conducted in the neonatal unit of my hospital. It revealed that this protein was the best marker of neonatal asphyxia. The analyzes were performed by nephelometry with reagents from Behring. Unfortunately, the technique on urine has not been "accredited" by the Behring. I have replaced it by cystatin C. New research with this molecule in neonatology has not been carried out.

    About kidney problems of  AIDS patients, monitoring can be easily realized on the basis of electrophoresis in SDS or determination of albumin and either low MW protein

  • Nicole Gillain-Martin added an answer in Blood:
    6
    How do you separate serum from blood without a centrifuge? or even without just letting it stand still for a few minutes?

    I don't want to use a centrifuge and the other method is time consuming. I wanted to know whether there is any other way wherein i can obtain serum from blood at home and instantly.

    Nicole Gillain-Martin

    Yes,  Soad Nasr, but the use of this kind of tube requires a centrigugation

  • Nicole Gillain-Martin added an answer in EDTA:
    14
    Is there any method to convert EDTA-plasma to serum?

    We have harvested a lot of discarded plasma samples from blood donor archive, which is containing EDTA as anticoagulant. Is there any method to convert EDTA-plasma to serum? Any idea is welcome. Thanks a lot! 

    Nicole Gillain-Martin

    I agree with Mr Banerjee. Obviously, this will only be possible if the component you want assayed does not depend on the presence of calcium (or Mg)  or is not complexed by EDTA

  • Nicole Gillain-Martin added an answer in Clinical Diagnostics:
    7
    A red crystal-like substance in urine. What can this be?

    I am sure this question is going to be one of the more peculiar ones ever asked here, so please forgive the strange atmosphere of the question! Being that I own my own laboratory and I wanted to learn more about urine (and it's components), I let curiosity get the better of me and decided to test whether or not i could separate urine from any of its components by gravity just for fun. Long story short, I collected my urine in a 50ml centrifuge tube and centrifuged it at various paramaters. Nothing happened during this time, however I was called suddenly away from the lab, and in a hurry, I mistakenly forgot to dispose of the urine in the tube before leaving! After returning about two and a half weeks later, I removed the centrifuge tube containing my urine from the rotor to find that there is a red crystal like growth on the wall of the tube.

    As I said, I do not know much about urine and I wanted to know if this was normal or not. What could the red crystal-like substance be? The centrifuge tube was sealed by the way and was never opened foe the entire time that it contained urine. The growth is quite visible and has the appearance almost identical to a naturally growing quarts or Amethyst crystal(but it is dark red). Any ideas what this is? Is is normal? Thanks!

    Nicole Gillain-Martin

    I would have also thought of urate that occur naturally in many urine stored in the refrigerator. Your urine has reached room temperature, so it is certainly alkalized.

    Think of ammonium urate and also to phosphate crystals

    To reassure you, ask a specialist to quickly carry out a review on a morning urine sample given in the shortest possible time in the laboratory!

  • Nicole Gillain-Martin added an answer in Valproic Acid:
    3
    What is the next test you would order for a patient with isolated ALP elevation?

    One of my patients, brought me an ALP=669 in an exam order by a psychiatrist because of the use of valproic acid (ALT, AST, GGT and Bilirubin are normal). I have already ordered serum Calcium, serum total protein and albumin, TSH and PTH. I wander if it is essential to order a bone scan.

    Nicole Gillain-Martin

    I fully agree with Mrs Poggiali,: it is essential to achieve electrophoresis of iso-PAL will specify hepatic or bone origine. Macromolecular could be present, but should not be responsible for such an increase without being accompanied by an increase of GGT.

    Still think a transient hyperphosphatasemia (very rare in adults) and check the persistence of high value

    This patient is not suffering from kidney failure, I suppose

  • Nicole Gillain-Martin added an answer in Antibody Generation:
    4
    Are serum levels of IgG, IgA or IgM antibodies generally known to change with age?

    In healthy individuals, do serum levels of any of the above immunoglobulin isotypes tend to increase or decrease? I can't seem to find any reference to them changing in a set pattern, but I may be looking in the wrong place/searching the wrong words..? Many thanks in anticipation..

    Nicole Gillain-Martin

    It is usual to consider that the immunoglobulins increases with age, but the data are sometimes conflicting. This is likely due to the observed differences based on race (white or black) and gender. Some factors must be considered such as smoking and alcoholism. The recruitment of the reference population can have a great influence on the conclusions of the study. Read about it: A. Gonzales-Quintela & coll: serum levels of immunoglogulins (IgG,IgA,IgM) in a general adult population and their Relationship with alcohol consumption, smoking and common  metabolic abnormalities, Clinical and Experimental Immunology, 2007, 151:42-50.

    In practice, immunosenescence does not cause a reduction of the production of immunoglobulins, but modifications of the "quality" of these immunoglobulins (lack of affinity, antibody activity)

  • Nicole Gillain-Martin added an answer in Metformin:
    17
    What is the cause of low hemoglobibh in 65 year old male with diabete?

    A 65 male patient with diabete has the following serum biochemistry results.

    blood glucose 120 mg/dl and taking glyclzide metformine and pitoz-Hemoglobin is 12g/dl which is low-serum iron is normal-serum ferritin is normal-RBC is 4.4,vitamine B12 is normal.What is the cause of Low hemoglobin.What is your suggestion for treatment for low hemoglobin?

    Nicole Gillain-Martin

    Does the patient have a chronic inflammatory syndrome, even discreet?
    (sedimentation rate, electrophoresis ...).

    Normal ferritin may correspond to an iron deficiency masked by this inflammation that will raise ferritin.

    It is also important to know the corpuscular volume and mean corpuscular hemoglobin concentration and the reticulocyte count.

    The patient's hemoglobin is slightly reduced and that is really worrying if you see in him a drop compared to previous values. Then seek occult blood loss

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