Kidney Function and Urinalysis

The excretory system consists of paired kidneys, a ureter draining each, a 
urinary bladder for accumulating urine and the urethra through which urine 
is voided.  (W&D pg 182)
Of the structures listed, only the kidney contribute to the production of 
urine.
Blood is supplied to the kidney via renal vessels.
(The term excretion refers to the release of urine from the kidney to the 
ureter.
Voiding of urine from the body thru the urethra is termed micturition.)

The kidney is a vital organ and humans cannot survive without at least one 
kidney.
The critical functions of the kidney include:
       1] the regulation of fluid volume and osmotic pressure;    
       2] the regulation of the chemical composition of iterstitial fluid, 
plasma and lymph
              a] metabolic wastes
              b] electrolytes
              c] pH

These functions are performed by nephrons, about 1 million/kidney, which 
are the functional units of the kidney.  (See text)
The kidney utilizes three processes to form urine:
        1] glomerular filtration
        2] tubular reabsorption
        3] tubular secretion

Filtration is accomplished due to the hydrostatic pressure of blood.  
Since glomerular capillaries are very permeable and the surface area is 
large, the glomerular filtration rate (GFR) of both kidneys averages 
180L/day (45 gallons)!  Obviously most of the water must be reabsorbed and 
returned to the vascular system to maintain blood volume and pressure.  In 
addition to water, the glomerular filtrate contains all small molecules 
found in plasma in essentially the same concentration (glucose, urea, 
amino acids, salts, water).  
Large molecules, such as proteins, and cells are not filtered.

As the filtrate travels through the nephron, most of its components are 
reabsorbed to varying degrees, depending on the need to adjust blood 
chemistry and the kidney's ability to reabsorb nutrients and other 
molecules.  The cells of the nephron may secrete ions and drugs as well.  
Since urine output is only 1 - 2 L/day, it is apparent that most of the 
water filtered, as well as required nutrients and electrolyes are returned 
to circulation.  The amount of urine produced varies with dietary intake, 
exercise and pregnancy.  

The liter+ of urine produced by a person contains about 60g of solids, 15g 
of which is NaCl.  Various organic and inorganic molecules contribute to 
the remaining 15g,  
including  creatine, amino acids, uric acid, ammonia, sulfates, potassium, 
and magnesium.  Occasionally and in pathological conditions, substances 
normally absent or present in trace amounts (such as glucose, proteins, 
hemoglobin, ketone bodies, cells, may be detected in the urine.   

Objectives:
1] To become familiar with the organization of the kidney
2] To examine urine samples for constituents 
3] To acquire an appreciation of urinalysis  as a diagnostic tool
4] To become familiar with the concepts of glomerular filtration rate and 
renal clearance


											        
Pg 2
I.  Kidney structure

    Examine a prepared slide of mammalian kidney.  Utilize W&D Fig 25.1 
and illustration    
        on chalkboard.   Identify:  
                      Gross features - cortex, medulla and medullary 
pyramid, pelvis
                      Microscopic features - renal glomerulus, Bowman's 
capsule, 
                                                              x-sec of 
convoluted tubules in cortex,
                                                              long.-sec of 
collecting tubules in medulla

   Answer the questions on the Lab Response Sheet.
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II. Urinalysis            W&D pg 182 - 185  no human subjects
 
     BioKit simulations and analyses.

     Because of the increased incidence of chemical abnormalities and 
health risks in human 
       populations, we will not sample our own body fluid.  Instead 
simulated specimens will be 
       provided.

     Follow instructions on the worksheet and especially pay attention to 
any modifications given 
        by your instructor.  Use liquids sparingly.

     Construct your own copy of the tables and fill in the data.  Answer 
questions from the 
       worksheet.

     Read the material in W&D on these topic.  Attempt to reason through 
the answers to W&D's 
     questions.

------------------------------------------------------------------------------------------------------------
III.  Glomerular Filtration Rate  
 
     To calculate GFR:    125 ml/min  X  60 min/hr  X  24 hr/day

                       daily  GFR =  180 L/ day  

A 70Kg person has a blood volume equal to about 7% body weight.  If we 
assume a hematocrit of 0.45,  we can calculate the plasma volume filtered 
each day.

    1 L/Kg  X  70 Kg  X  0.07 (1 - 0.45) = plasma volume = 2.695 L plasma

If the kidneys filter 180 L of plasma per day and the plasma volume is 2.7 
liters,
the total plasma volume is filtered ____________ per day.       

Answer question on Lab Response Sheet.
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IV.  Renal Clearance

To characterize the combined functions of filtration, reabsorption and 
secretion, the concept of renal clearance is employed.  If you know the 
volume of plasma that would contain the amount of substance appearing in 
the urine per unit time you can measure kidney function.  This volume of 
plasma is known as the renal clearance of that substance.

To assess this value, we can choose a substance that can be readily 
determined quantitatively, is completely filtered by the glomerulus, and 
is physiologically inert (neither absorbed not secreted).
Inulin is polysaccharide that satisfies these conditions.  

    So,      Pin  =  plasma conc. of inulin  (mg/ml) ,

               Uin   =  urine conc. of inulin  (mg/ml),

                V    =  urine flow ml/min, and
                Cin  =  plasma clearance of inulin

The clearance is related to these parameters as:

                Cin  =  V x Uin / Pin           and also equals GFR.

The clearance value for inulin is about 135 ml/min.

The comparison of renal clearances of several substances can be made to 
that of inulin.
For example we find that the renal clearance of p-aminohippuric acid (PAH) 
[widely used as diagnostic drug in renal disease]  is 650 ml/min.   Since 
this is well above the GFR  and renal clearance value for inulin, it is 
reasonable to  conclude that PAH is being secreted into the tubule as well 
as being filtered at the glomerulus.  
On the other hand, glucose normally has a clearance of zero.  If plasma 
concentration of glucose exceeds the capability of the reabsorption system 
of the nephron, then glucose would appear in the urine.
Since urine production is only a fraction of the filtration rate, it 
should be apparent that the bulk of the glomerular filtrate is reabsorbed 
by the nephron.  Experiments show that  85% of the  glomerular filtrate is 
absorbed by the proximal convoluted tubule.  The remaining fluid may be 
further modified to produce urine.  An impressive fact - humans can 
maintain salt and water balance on less than a half a gram of salt and a 
liter of water a day.

W&D pg 182 - 185  no human subjects
 
     Read the material in W&D on this topic.
                                                                                 
     Think about the questions posed by W&D.  Answer the questions on the 
LRS.













Lab Response Sheet

I.  Kidney Structure.

    1]  Where are most of the glomeruli located in the kidney?

    2]  What is the function of the glomeruli?

    3]  Using your text (homework) list, in sequence, the parts of the 
nephron through which the 
         filtrate passes.

II.  Urinalysis -  construct your own tables for your group's data; answer 
the questions 

III. GFR
    4] A "normal" woman's kidney filters, on average, 115 ml/min and 
hematocrit is 38%.
        Ms. Muffit  weighs 70 Kg  but suffers from hypertension.  Due to 
her high blood pressure,
        her  kidneys are filtering 135 ml/min. 
        Calculate the normal GFR for women.

        Calculate Ms. Muffit's GFR and plasma volume filtered per day.

        Adjusting for gender, how much more work, as % normal, is Ms. 
Muffit's kidneys doing?

IV.  Renal Clearance
      5] Define secretion and reabsorption




      6] If we had done the comparison among your classmate described in 
W&D,  what variables 
           could have affected the values obtained for different 
individuals?


      7] What is the main solute in blood?
          Also in urine?












Notes to TAs:
I will make 15 copies of the handout.  Please do not let them "walk."  
Students may download the handout from my web site.  If you want them to 
turn in anything at the END of lab then make copies of the Lab Response 
sheet.

I will put 1 or 2 diagrams on the wall chalk board.  You may wish to use 
them in your introduction.  

Kits have been ordered and materials to supplement kits or to replace a 
theses are utilized.  Please be frugal so that we don't run out at end of 
wk.

Although work with the kits is the main focus, students should look at 
kidney and you should talk students thru W&D.