KERMES 



170 



KIDNEY 



dermis (Favre, M. Ann. de Dermat. et 

 Syph., 1950, 10 (3) , 241-262) . The litera- 

 ture 1886 to date includes reports of the 

 origin of keratohyalin granules from 

 nuclear substance, intercellular fibrils, 

 and cytoplasm. 



The particulate nature of the granules 

 has been demonstrated recently by their 

 isolation from epidermis (D. L. 

 Opdyke). Epidermis is separated by 

 the heat method of Baumberger, J. P., 

 Suntzeff, v., and Cowdry, E. V., J. 

 Nat. Cancer Inst., 1942, 2, 413-423. The 

 sole of the foot offers an ideal place to 

 obtain keratohyalin granules. 



After separating the epidermis from 

 the dermal layers, the proximal side of 

 the epidermal sheet is scraped with a 

 knife, removing all of the epidermis in 

 fine shavings down to the highly trans- 

 parent, thick stratum corneum. The 

 shavings are then homogenized in a 

 Waring Blendor until no cells remain 

 intact. This homogenate is made in a 

 0.85% NaCl solution of pH 7.3 to which 

 a crystal of thymol is added to minimize 

 bacterial contamination. All proce- 

 dures are carried out in a cold room. 

 The process of cell fractionation in the 

 micro cup of the Waring Blendor re- 

 quires about 45 min. If 2% citric acid 

 is employed as the medium, this time 

 may be reduced to 15 min. 



The components of the suspension are 

 then separated by differential centrif- 

 ugation. Nuclei and whole cells are 

 found to sediment at 2.5 to 3 Kg/g. 

 The keratohyalin granules sediment at 

 25.2 Kg/g. after 12 min. of centrifuga- 

 tion. These can be resuspended in 

 fresh saline, washed, and reprecipi- 

 tated. They can be distinguished from 

 mitochondria by their tinctorial 

 properties. 

 Kermes. This scarlet dye was known in 

 Egypt and farther East at a very early 

 date. Kermes is the Armenian term 

 for a "little worm", variously identi- 

 fied as Coccus arborum and Coccus ilicis. 

 Moses referred to it as "Fola" and 

 "Fola shami". Remember the promise 

 of Jehovah: "Though your sins be as 

 scarlet (Fola) they shall be as white as 

 snow; though they be red as crimson 

 (Fola shami), they shall be as wool". 

 So valuable was Kermes that after the 

 subjugation of Spain by the Romans 

 the people were made to pay half of the 

 tribute in Kermes. At about 1640 a 

 Dutch chemist discovered the similarity 

 of this dye to cochineal. Its history 

 affords interesting reading (Leggett, 

 W. F. Ancient and Medieval Dyes. 

 Brooklyn: Chemical Publishing Co. 

 Inc., 1944, 95 pp.). 

 Kidney — Written by Jean Oliver, Dept. of 



Pathology, State University of New 

 York, Brooklyn 2, N. Y., September 4, 

 1951 — Techniques for the general 

 demonstration of the elements of the 

 renal tissue, epithelial cells, sustaining 

 tissues, blood vessels and nerves are 

 essentially the same as those used for 

 other organs. Masson's Trichrome 

 stain has the advantage of affording a 

 particularly colorful differentiation of 

 the various elements in a single section. 



The individual renal organs that make 

 up the kindey, the nephrons, can be 

 isolated in their entirety by maceration 

 and teasing as described by Huber, G. 

 C, Cowdry's Special Cytology, 1932,2, 

 935-977. Partly wash out blood by in- 

 jecting physiological saline into the 

 renal artery. Then follow with hydro- 

 chloric acid (cone. HCl, 3 parts and aq. 

 dest. 1 part) using care to protect the 

 eyes. Remove and immerse the organ 

 in the same fluid. After a suitable time, 

 determined by excising pieces, wash a 

 block of tissue with aq. dest. and stain 

 in Hemalum. Wash in very dilute aq. 

 sodium hydrate. Isolate individual 

 tubules by teasing with fine needles. 

 Wash, and mount in glycerin. With 

 small mammals Ruber's results were 

 excellent but he was not satisfied with 

 his human preparations. The method 

 has however been well adjusted to the 

 human kidney by Oliver, J., Architec- 

 ture of the Kidney in Chronic Bright's 

 Disease, New York: Paul B. Hoeber, 

 1939, by a simpler procedure and dis- 

 sected nephrons may be mounted, 

 stained and photographed. A method 

 of montage then affords a demonstra- 

 tion in their natural continuity of the 

 cellular elements of the nephron at 

 high magnification (J. Clin. Invest. 

 1951, in press). 



A clear distinction between glomeruli 

 and the renal tubules is important. It 

 is a simple matter to color the former 

 with 1:5000 Janus blue (which is more 

 satisfactory for this purpose than Janus 

 green) in 0.85% aq. sodium chloride by 

 vascular Perfusion and to determine 

 their number, size and distribution 

 against a background of unstained or 

 faintly rose tinged tubules in slices of 

 fresh kidney (Cowdry, E. V., Contrib. 

 to Embryo! . Carnegie Inst., Washing- 

 ton, 1918, 8, 39-160). 



Perhaps in no other organ is it pos- 

 sible to correlate morphological struc- 

 ture with functional activity so closely 

 and by so many methods as in the kid- 

 ney. See Oliver, J., Am. J. Med., 

 1950, 9, 88 for a general statement of 

 the problem with various examples of 

 such procedures. As an example, the 

 technique for the microscopic study in 



