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Alkaline Information and Courses from MediaLab, Inc.

These are the MediaLab courses that cover Alkaline and links to relevant pages within the course.

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Laboratories Individuals

Chemical Screening of Urine by Reagent Strip
False-positive tests for protein may be caused by: (Choose ALL of the correct answers)View Page
What affect may bacterial contamination have on urine pH?View Page
Which of the following statements are TRUE for specific gravity measured by the reagent strip method? (Choose ALL of the correct answers)View Page
Basis of the Urine pH Test

Dipsticks measure pH using methyl red and bromthymol blue indicator dyes. The color change that occurs in this test area correlates with the urine's pH. Sensitivity to pH ranges from 5.0 (acid pH) to 9.0 (alkaline pH) on a urine reagent strip.

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Acid and alkaline urine pH

Reasons for acidic urine pH include: a high-meat diet, respiratory/metabolic acidosis, and hypochloridemia. A urine with a high concentration of glucose may also have a lower pH. An alkaline pH may be the result of a vegetarian diet, respiratory/metabolic alkalosis, or a bacterial infection caused by urease-producing bacteria. Urine that contains bacteria can become more alkaline if the specimen remains at room temperature for an extended period of time.

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Urine Specimen

The urine specimen should be freshly voided. Urine is an ideal medium for the proliferation of bacteria due to the large amount of urea present. These bacteria metabolize urea, producing ammonia that increases the urine pH. If there is a delay before performance of the test, the sample should be refrigerated. This will: Prevent urease-producing organisms, such as Proteus and Pseudomonas, from converting urine urea to ammonia, which results in an increased pH. Prevent loss of CO2 which increases pH to the alkaline range.The "run-over" phenomenon may occur if excess urine remains on the strip. The protein area, adjacent to the pH area, contains an acid buffer which may "run-over" the pH portion resulting in an acid reading on a neutral or alkaline urine.

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A urine specimen was collected at 6:00 A.M. and remained at room temperature until it was received in the laboratory at 3:30 P.M. How may the pH of the specimen be affected by the extended time at room temperature if bacteria are present in the specimen?View Page
Match the following factors with the expected pH:View Page
False Positive Protein Results

A urine specimen that has remained at room temperature for an extended period of time may produce a false-positive protein result on a reagent strip. A false positive may also occur in the presence of bacterial contamination, alkaline medication, or quaternary ammonium compounds such as disinfectants or drugs, and with skin cleansers containing chlorhexidine.

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Testing for Ketone Bodies

Testing for ketone bodies is based on a nitroprusside reaction. Acetoacetic acid reacts with sodium nitroferricyanide and glycerine in an alkaline medium to produce a violet-to-purple colored complex. The reagent strip method can detect as little as 5 mg/dL acetoacetic acid in urine. It does not react with acetone unless glycine is present or B-hydroxybutyric acid. Since these two compounds are derived from acetoacetic acid, their presence can be assumed if the test for ketones is positive. Ketones are reported either as negative, small, moderate or large amounts; or negative, 1+, 2+, 3+, or 4+. In some severe cases of ketosis, it may be necessary to perform tests on serial dilutions to provide more information on the quantity of ketones present.

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Diazotized 2, 4, dichloroaniline reacts with bilirubin in an ___________ medium.View Page
Advantages and Limitations of the Reagent Strip Method for Specific Gravity

Specific gravity measured with the reagent strip method correlates well with gravimetric measurement, and, unlike the gravimetric or refractometer methods, does not need to be corrected for glucose or protein. Cloudy/turbid urines do not need to be clarified before measuring specific gravity with the reagent strip method. It is the recommended method for determining specific gravity if a urine specimen contains x-ray contrast media or plasma expanders. Alkaline urine can affect the indicator system and lower the specific gravity result on the reagent pad. If the result is being read visually, it is recommended that .005 be added to the result when the pH is alkaline. Most dipstick readers, however, will automatically adjust the specific gravity reading for pH. A specific gravity reading higher than the reagent strip range would need to be measured by another method, and may require dilution.

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A visually-read specific gravity on a urine specimen with an alkaline pH should be adjusted by:View Page

CLIA Chemistry / Urinalysis Review
Which one of the following crystals is not found in normal urine:View Page
Identify the urine sediment elements indicated by the arrow in the illustration:View Page
Identify the sediment elements present in this alkaline urine:View Page
Serum alkaline phosphatase activity is derived from all of the following organs except:View Page
Alkaline phosphatase activity is increased in all of the following conditions except:View Page
All of the following are sources of serum alkaline phosphatase except:View Page
Which of the following conditions would be suggested by a marked rise in alkaline phosphatase, jaundice, and a moderate rise in ALT:View Page

CLIA General Laboratory Review
The normal range for urine pH is:View Page
Match the urine sediment or crystal to the correct description.View Page
The Kleihauer-Betke test is used to:View Page

CLIA Hematology / Hemostasis Review
The procedure which may be used to assist in differentiating chronic myelocytic leukemia from leukemoid reaction is:View Page

Confirmatory and Secondary Urinalysis Screening Tests
Screening and Secondary Tests for Protein

A routine reagent strip protein method, based on the principle of "protein error of indicators," produces a visible colorimetric reaction that is capable of detecting most instances of proteinuria.Traditionally, laboratories have used sulfosalicylic acid (SSA) to confirm all positive protein reagent strip results, but this practice is not as common in today's laboratories. SSA is a precipitation method that reacts with all forms of protein. However, any substance that is precipitated by acid will produce false-positive SSA results. This includes radiographic dyes, cephalosporins, penicillins, and sulfonamides. SSA may be used as a secondary protein detection method if the urine is highly alkaline (pH of 9.0 or greater) which would overwhelm the buffering capacity of the reagent on the protein reagent stick. SSA may also be used as an alternative protein detection method if the urine is highly colored so that the colorimetric reaction is masked on the reagent strip.

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Pharmacology in the Clinical Lab: Therapeutic Drug Monitoring and Pharmacogenomics
Protein Binding

Most drugs are bound to proteins when they circulate in the body. Albumin is a major drug-binding protein in serum. Albumin is an alkaline protein, so acidic and neutral drugs primarily bind to it. If albumin binding sites become saturated, acidic and neutral drugs can bind to lipoproteins. Alkaline drugs tend to bind to globulins, particularly to the globulin, alpha-1 acid glycoprotein. Only free, unbound drugs are able to bind drug receptors and have therapeutic effects. An equilibrium exists in the systemic circulation between a free and protein-bound drug and between a free and receptor-bound drug. This is illustrated in the image to the right.

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Chemiluminescence

Chemiluminescent assays use antibodies that are conjugated to enzymes, such as peroxidase or alkaline phosphatase. These enzymes, mixed with chemiluminescent substrates, produce light in the visible spectrum. A direct relationship exists between the amount of drug that is present in the sample and the light units that are produced and measured by the luminometer in the instrument. Assays that use chemiluminescence are more sensitive than immunoassays that rely on the generation of a colored product.

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Phlebotomy
Hepatic function panel

Albumin (Alb) Bilirubin (Bili) Alkaline phosphatase (Alk Phos) Total protein (TP) Alanine aminotransferase (ALT) Aspartate aminotransferase (AST)

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Comprehensive metabolic panel (CMP)

Consists of a basic metabolic panel, plus:Albumin (Alb) and Bilirubin (Bili) Alkaline Phosphatase (Alk Phos) Total protein (TP) Alanine aminotransferase (ALT) Aspartate aminotransferase (AST)

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The Urine Microscopic: Microscopic Analysis of Urine Sediment
Use the following urinalysis report to answer:The patient was a female and the examination was completed within two hours of collection. Color - light yellow Appearance - slightly turbid Sp. Gravity - 1.009 pH - 8 Glucose (Multistix) - 0 Glucose (Clinitest) - 0 Protein - 1+ Blood - 0 WBC - 5/HPF RBC - 1/HPF Epithelial - 0/HPF Casts - 2 hyaline/LPF Crystals - amorphous urates Bacteria - 2+True or false? The results are abnormal but all results correlate.View Page
Urine Sediment

Urine sediment may also contain white blood cells (WBCs). Most of the WBCs in urine are segmented neutrophils. Since it is possible that lymphocytes, monocytes, and/or eosinophils may be present, the cells in urine can be stained if it is necessary to differentiate them. The segmented neutrophil just above center of the image to the right shows a distinct nucleus. When viewing urinary sediment under the microscope, the fine focus adjustment must be used to identify white blood cells. White blood cells swell in dilute alkaline urine and the cytoplasmic granules exhibit brownian movement resulting in “glitter cells.” These cells lyse rapidly. “Glitter cells” are most easily seen when viewed under phase-contrast microscopy.

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Swollen RBCs

In contrast, RBCs appear swollen in dilute or alkaline urine, having taken on water from their surroundings.

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Amorphous Urates

Amorphous urates appear as dark or yellow red granules while phosphates are white or colorless. The pH of the urine determines the type of amorphous crystals present. They may be urates in acid urine or phosphates in alkaline urine.

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Crystals in Normal Alkaline Urine

Crystals found in normal alkaline urine include triple phosphate, ammonium biurate, calcium carbonate, amorphous phosphates, and calcium oxalate. This slide shows an example of triple phosphate crystals. These may appear as four to six sided prisms resembling coffin lids. They indicate either stasis of the bladder or a stale sample.

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Which of the following may be found in normal alkaline urine?View Page
Please identify these crystals occurring in acid urine.View Page

White Cell and Platelet Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
Typical cells on a peripheral blood smear as photographed here were repeatedly encountered as the smear was reviewed. The peripheral white blood cell count was 51,000/ml with an orderly maturation sequence. The comment "leukemoid reaction" may properly be appended to the report.View Page
Leukemoid reaction revisited

The term leukemoid reaction is used to describe peripheral white blood cells that on the stained blood smear may have some resemblances to leukemia cells. Quantatively in a leukemoid reaction, the neutrophil count is >50,000 cumm with more immature cells, particularly myelocytes, than are usually present in toxic left shift syndromes. The presence of immature cells in a leukemoid reaction awakens thoughts of leukemia. Great care must be taken to make a distinct differentiation between aberrant white blood cell proliferations and a benign but exaggerated granulocytic proliferative response. Our material is from a 1-month-old girl with Down's syndrome. Her total white blood count was 37,000/mm3 interpreted as leukocytosis with left shift. Leukocytosis with a left shift, and leukemoid reactions with high alkaline phosphatase are conditions to be mindful of in patients with Down's syndrome. The alkaline phosphatase score is high in leukemoid reactions, low in granulocytic leukemia.

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A most useful follow-up test to consider when faced with hypersegmented neutrophils and oval macrocytes (see photograph) in a peripheral blood smear is:View Page


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