Morphogenesis of calcium laden cytoplasmic bodies in malakoplakia of the skin

MORPHOGENESIS OF CALCIUM LADEN CYTOPLASMIC BODIES IN MALAKOPLAKIA OF THE SKIN An Electron NIicroscopic Study Harold k1. Price, lH.D.,* James B. Hanrahan, M.D.,t and Robert G. Florida, B.S.+

Abstract Thc origin and dcvclopment ofthc ~lichaelis-GlIltmannbody arc presclllcd in a casc of malakoplakia of thc skin from which coagulasc positivc S/a!J/ty!ococCllS llllfellS was culLurcd. Elcctron microscopic obscnations indicatc an ordcrly scquencc of c\'cnts that rcsulLs in thc formation of Michaelis-Gullmann bodics within singlc mcmbranc limitcd cytosomes in thc cytoplasm of histiocytcs. Thc wcll dcveloped ~lichaelis-GlItLInann bodies are composed of closely packed, gcnerally radially oriented, needle shaped, crystalline deposits with lengths varying from 400 to 1450 A and widths from 60 to 120 A. Selectcd area electron diffraction studies of the ~Iichaelis-Gullmannbodies show a diffraction pallern consistent with the x-ray powder diffraction pattern for hydroxyapatitc.

The most distincti\'c histologic fcaturc of malakoplakia is thc prcscncc of imraccllular and intcrccllular mincralized sphcrulcs known as ~lichaelis-GutLIllann bodics. Thcy arc associatcd with thc pre\'ailing shccts of histiocytic cclls charactcristic of thc "classic phase" of this rarc discasc. 1 The Michaelis-Gullmann bodics arc PAS positivc, diastasc rcsistant inclusions that stain posith'e1y when thc von Kossa and alizarin red S calcium rcactions and thc Perls iron stain arc used. I - 3 They havc a concclllric lamcllar pallcrn in hoth light and electron microscopic prcparations. l. 3. 4

Preliminary x-ray diffraction analysis has indicatcd a calcium phosphatc complex, such as thc apatitc found in bonc and thc psammoma bodics of thyroid and ovarian neoplasms. I From ultrastructural studics, sevcral workcrs havc suggcstcd that ~Iichaelis-Gullmannbodics may rcprcsent mincralizcd cytosegrcsomcs.3 • 4 Howcvcr, no sUlTounding limiting mcmbranc was sccn, and a transitional stagc betwecn thc typical mcmbranc bound cytosegl'esomcs of thc histiocytcs and thc I\lichaciis-Gullmann bodies could not bc dcmonstratcd. U mil 1958, when a case with involve-

*Associatc

Profcssor of Pathology. Unh'crsity of Pittsburgh School of ;\Icdicinc. Pathologist. ;\Iagcc-Womcns Hospital, Pillsburgh, Pcnnsyh·ania. tClinical Assistalll Profcssor of Pathology. Unh'crsit}' of Pillsburgh School of ;\Icdicinc. Patholob';st, Thc Wcslcrn Pcnns}'h'ania Ilospital. Pillsbllrgh, Pcnns}'!vania. :l:Rcscarch Assistatll. Electron ;\Iicroscopy Labor-lIor}', ;\Iagcc-\\'omcns Hospital, I'illsbllrgh. Pcnnsylvania.

381

HU~IAN

PATHOLOGY-VOLUME 4,

NU~mER

ment of the testes was reported,i malakoplakia was thought to affect only the mucosa of the bladder, ureters, and renal pel\'is. Subsequent reports ha\'e affirmed its presence in the prostate,i' 9 kidney,IO-12 retroperitostomach,13 colon,2' 5.13-16 neum,2 lung,12 and boneP In addition, the entity "megalocytic interstitial nephritis" has been referred to as a form of malakoplakia limited to the renal cortexY The·' etiology has not been established, but it is generally associated with a local chronic inflammatory process, frequently an Escherichia coli infection. l . 2 It has e\'en been proposed that malakoplakia be regarded as a part of the spectrum of xanthogranulomatous inflammation.3 This paper reports, to our knowledge, the first example of malakoplakia of the skin as well as the first definiti\'e ultrastructural e\'idence that ~Iichaelis-Guttmann bodies e\'ol\'e inside single membrane limited cytoplasmic inclusion bodies of histiocytes. CLINICAL DATA Tissue was obtained from an ulcerated skin lesion, 3.0 cm. in diameter, of the midsternal region of the chest of a 62 year old white male (Fig. I). The patient had first become aware of the lesion six months prior to the admission to the hospital. The skin process began as a small red area, which gradually enlarged and began draining clear fluid and later small amounts of purulent material. At the time of biopsy the ulcerated region had some whitish exudate and granulation tissue within it. Laboratory studies included: a hemoglobin le\'e1 of 12.8 grams; hematocrit, 40; white blood cell count, 6500 per cu. mm. with a normal differential; cholesterol, 198 mg. per 100 ml.; calcium, 9.3 mEq. per liter; inorganic phosphate, 3.9 mg. per 100 ml.; total bilirubin, 0.7 mg. per 100 ml.; serum albumin, 3.5 mg. per 100 ml.; Figure 1.

3

September 1973

uric acid, 4.8 mg. per 100 ml.; blood urea nitrogen, 14 mg. per 100 ml.; glucose, 115 mg. per 100 ml.; serum lactic dehydrogenase, 129 international units per ml.; alkaline phosphatase, 16 King-Armstrong units; serum glutamic oxaloacetic transaminase, 35 units per ml.; and prothrombin time, 100 per cent of normal. Urinalysis was entirely within normal limits on two occasions. A culture of material taken from within the chest wound grew out coagulase positi\'e StajJ!tylococClls allrells. Cultures for fungi and acid fast organisms were negati\'e and guinea pig inoculations were also negath'e for tuberculosis. X-ray \'iews of the chest wall did not show any im'ol\'ement of the sternum or underlying ribs, but they did re\'eal changes consistent with fibrosis and emphysema throughout both lung fields. An electrocardiogram demonstrated e\'idence of an old anterior wall myocardial infarction and ST-T wa\'e abnormalities compatible with coronary artery disease. The lesion was widely incised, the incision carried deep to where a tract was found, which extended through the subcutaneous tissue of the chest wall but did not in\'ade the fascia or the bone. Se\'eral pieces of necrotic tissue were remo\'ed. All granulation tissue was also excised, lea\'ing the wound wide open and clean. The wound was packed with iodoform gauze. The patient's postoperati\'e course was uncomplicated, and the wound subsequently healed with no complications and no e\'idence of any further disease. The patient has been followed for almost a year with complete healing of the skin lesion.

MATERIALS AND METHODS Tissue from the margin of the ulcer was fixed in 2 per cent glutaraldehyde buffered with ~Iillonig's buffer,ls or in

Ulcerated 3 em. skin lesion in midsternal region.

Figure 2. Dense cellular infiltrate composed primarily of histiocytes at base of ulcer depicted in Figure I; arrow indicates margin of ulcer. (Hematoxylin and eosin stain. X 34.) Figure 3.

Histiocytes containing Michaelis·Guttmann bodies (arrows). (Hematoxylin and eosin stain.

Figure 4.

Calcospherules (:-'IG bodies) of varyil:g size. (Von Kossa stain. X 160.)

X 550.)

382

MALAKOPLAKIA OF THE SKIN-PRICE Figure I

Figure 3

ETAL.

Figure 2

Figure 4

383

HU~IAN

PATHOLOGY - VOLUME 4, NUMBER 3

buffered formalin soltllion, and processed for light and electron microscopy. The light microscopic sections were stained with hematoxylin and eosin, periodic acid-Schiff stain with and without diastase, \'on Kossa and alizarin rcd S for calcium, Masson trichromc, Perls' iron mcthod, acid fast, and mcthcnaminc sil\'cr stains. Thc tissue fixed in glutaraldchydc for elecu'on microscopy was postfixcd in 2 pcr ccnt ~rillonig's buffcrcd osmic acid,18 and cmbcdded in at'alditc.J!' Scctions 1.5 to 2 microns thick were cut and stained with toluidinc blue 20 or mcthylene blueazure 11.21 FollO\\'ing a light microscopic sur\'ey of "thick sections" thc blocks were trimmcd and thin scctions mountcd on grids for electron microscopy. The grids wcre examined both unstaincd and after staining with uranyl acctatc 22 and \cad citratc,23 some beforc and aftcr decalcification with 5 per cent acetic acid. For selected area electron diffraction studies, a Phillips EM 300 electron micro,scope was used with an accelerating \'oltage of 100 h. The diffraction camcra constant, LA, was dctcrmincd by using a diffraction pattern composcd of sharp rings obtained by photographing a dispcrsion of aluminum crystallitcs depositcd onto a carbon substrate. Then thc d \'alucs of the ~richaelis-Gutunann bodics wcrc calculatcd from diffraction patterns by using a \'ariation of the Bragg cquation, d =

~,

in which thc d \'alucs arc expressed in Angstrom units. 24 OBSERVj\.TIONS Light Microscopy

Thcre was focal ulceration of thc cpidermis exposing the undcrlying dermis, which contained sheets of histiocytic cells

Sejl/ember 1973

that extended laterally and downward into the underlying subcutaneous adipose tissue of the hypodermis (Fig. 2). The histiocytic cclls were embedded in a scant fibrous stroma containing \'arying numbcrs of plasma cclls, Iymphocytcs, and occasional polymorphonuclear Icukocytcs. Round to o\'al, hcmatoxylin staincd cytoplasmic inclusions \'arying from 4 to 12 microns in diametcr were seen in the histiocytcs, and occasionally entrapped bctwccn cclls. Thcse inclusions wcre either homogenous in appearance or had thc charactcristic target shaped pattcrn of thc ~[jchaelis-Guttmann body (Fig. 3). Thcy appearcd to bc periodic acid-Schiff positi\'c (with and without diastase) and staincd for calcium with the \,on Kossa and alizarin red S reactions (Fig. 4). Pcds' iron reaction was also generally positi\'e. Thc conccntric laminated pattcrn of thc Michaclis-Guttmann bodics was oftcn more striking in the periodic acid-Schiff and iron stained sections.

Electron Microscopy

Thc ultrastructural observations arc essentially restricted to the t)·pcs of single membranc limited cytoplasmic inclusion bodics seen in the histiocytes and the apparcnt relationship between thcsc cytoplasmic particles and the Michaelis-Guttmann bodies. As se\'eral re\'iewcrs ha\'c indicated, the nomenclature of cytoplasmic bodies has become complex and is in an almost constant state of flux. 2:;. 26 From the \'arious proposed classifications, it was cOl1\'cnient to adopt the terminology of Trump and Ericsson 2:; in dcscribing the 'cytoplasmic inclusions for this prcsentation. CYTOSO:\IES. Thc cytoplasm of the histiocytcs contained numerous round, o\'al, or polygonal cytosomcs gcncrally

Figure 5. Multiplc cytosomes and single calcosphcrule in C)'toplasm of histiocytc. (Uran}'1 acetatc and Icad citratc stain, X 17,100,) Figure 6, ~'ultiplc C)'toSOll1CS Idth intcrnal mcmbranous lamcllac. In thc C)'tosomc on thc far right thc membranes appear to bc arrangcd in a circular pattcrn of conccntric la}'crs. (Uran}'1 acctatc and Icad citrate stain. x 38,800.)

384

Figure 7. ~lcmbranol1s lamcllac, in a C)'tosomc, that consist of unit membranes hal'ing a thickncss of approximately 90 A. (Ur:lny' acctate and lead citrate stain, X 292,000.)

~IALAKOPLAKIA OF THE SKIN -PRICE ET AL.

Fi

ure

7

385

HUMAN PATHOLOGY -: VOLU~IE 4, NUMBER 3 September 1973

limited by a single membrane (Figs. 5, 6). Occasionally the limiting membrane appeared to be interrupted, probably as the result of processing artifact or plane of section. Internally they contained a finely granular electron dense material in which groups of stratified membranes were frequently found (Figs. 5 to 7). The membranous strata generally appeared to be arranged in semicircular fragments, but on occasion assumed a myelin-like configuration of concentric layers. A number of the cytosomes contained small oblong dense particles, which were either randomly scattered or had assumed the' layered pattern characteristic of the MichaelisGuttmann body (see below). By selected area electron diffraction, these needle shaped structures generally shO\\"ed a crystalline pattern resembling that of hydroxyapatite, and in large clusters they probably would yield the positi\'e von Kossa (Fig. 4) and alizarin red S reactions seen by light microscopy. CYTOSEGRESO:\IES. The cytoplasm of the histiocytes also contained occasional single membrane limited bodies, which were packed with recognizable organelles, such as mitochondria, endoplasmic reticulum, lipid droplets, and even cytosomes (Fig. 11). Sometimes they contained nondescript dense membranous components and finely granular material. A few of these inclusions had degenerated organelles as well as membranous and granular matrix material suggesting a transitional state between a cytosome and cytosegresome (Fig. 12). ~hCHAELIs-GuTnIA~~ BODIES. Ultrastructurally the ~Iichaelis-Guttmann bodies appear to be spherules composed of alternating dense and less dense layers of crystalline deposits (Figs. 9, 10, 13, 15,

16). Some of the Michaelis-Guttmann bodies have a dense core followed by a less dense zone, in turn followed by a denser zone; or vice versa, a less dense core followed by a denser ring, and so on. Occasionally only a dense ball of crystalline material was seen with no layering effect (Fig. 5). Further sectioning sometimes indicated that this was an outer dense zone of a spherule. Small satellite spherules were sometimes seen, resulting in a pseudobudding effect (Fig. 15). Following decalcification a homogenous, less dense, noncrystalline matrix material was evident (Figs. 13, 14). The ~Iichaelis-Guttmannbodies were seen both intracellularly and extracellularly (Fig. 16). In the intracellular form they were generally enclosed in a single membrane lined structure resembling a cytosome (Figs. 9, 10, 15). In some cytosomes only randomly scattered, dense particles could be seen (Fig. 8), and in others a transition to the more ordered lamellar pattern of the filichaelis-Guttmann body was evident (Figs. 9,10,15). Occasionally free nonmembrane limited clusters of crystalline material were seen within the cytoplasm (Fig. 17). The crystalline deposits generally had a needle shaped configuration with lengths varying from 400 to 1450 A and widths from GO to 120 A. In thinner sections they often appeared to be oriented in a radial direction in the filichaelis-Guttmann spherules (Fig. 16). By selected area electron diffraction they gave a pattern similar to that of hydroxyapatite (Table 1; Fig. 19). In those cytosomes with dense particles that generaHy lacked a distinct needle-like pattern no rings were seen when they were subjected to selected area diffraction (Fig. 8). This latter finding may be a result of

Figure 8. Dense "calcific" particles scattered within the matrix material of a cytosome; see text. (Uranyl acetate and lead citrate stain. X 22,600.) Figure 9. Possible early organization of calcospherule (MG body) \\'ithin cytosome; see text., (Uran}'1 acetate and lead citrate stain. X 20,400.) Figure 10. Calcospherule (MG bod}') with core containing relatively sparse aggregates of crystalline material and two dCII~CI' outer lamellar rings containing abundant aggregates of apatite crystals. Arrows indicate membranous margin of cytosome in which this ~IG body is located. (Uranyl acetate and lead citrate stain. X 17,900.)

386

Figure 1 I. Cytosegresome containing multiple cytosomes (c), mitochondria (m), and fragments of endoplasmic reticulum (arrow). Clear spaces probably represent zones where material of lipid droplets was dissolved out during tissue processing. (Uranyl acetate and lead citrate stain. X 17,300.)

MALAKOPLAKIA OF THE SKI1\' -PRICE Figure 8

Figure 9

Figure 10

Figure II

ET AL.

387

HUl\IAi\' PATHOLOGY - VOLUl\1E 4, NUMBER 3

Fi

r

SejJtemher 1973

12

Figure 12. Large cytoplasmic inclusion that contains a dcgcncrati\'c mitochondrion (m) and myclin-likc figme (ml) within a matrix of find)' granular electron dense matcrial. (Uranyl acctate and lead citratc stain, X 40,000.) Figures 13 and 14. Serial sections of same calcospherulc before (Fig. 13) and aftcr decalcification (Fig. 14). (Unstained. X 13,700.)

388

the presencc of amorphous prcCl"ystalline mincralizcd componcnts, but it is morc likely that thc dcposits of dcnsc matcrial at this early stagc wcrc too small to be elfectiycly cyaluated by elcctron diffraction. 24 Thcre is no morphologic cyidcncc of cndocytic or cxocytic actiyity; and it is di/Iicult to cxplain thc prescncc of cxtraccllular ~Iichaclis-Guttmann bodies or e\'cn how thc ccll handles thc occasional c1ustcrs of nonmcmbranc bound intracytoplasmic Cl"ystallinc matcrial.

With the Pcrls iron reaction onc would expect to sec fcrritin deposits within the ~Iichaclis-Guttlllann bodies. In the elcctron micrographs. howcycr. it is not easy to detcct ferritin deposits unlcss thcy are part of the denser granular matcrial disperscd among the hydroxyapatite crystals or arc possibly adsorbed to the shell of thc hydroxyapatite crystals. H ,47 In some of the cytoscgresomes dense dcposits resembling hcmosiderin granulcs are scenP OTHER CELLULAR ORGA:'\ELLES.

ccpt for occasional degeneratiye

Ex-

chan~es

l\IALAKOPLAKIA OF THE SKIN-PRICE

ET ,\L.

Figure 15. Cytosomcs contanllng :\lichadis-(;lIttmann Lodics in various stagcs of dc\"elopmclll" Small C)losome to the right of celller (arrow with a c) contains randomly scattered. small. dcnse "calcific" paniclcs as seen in Fib'Ure 8. Large lowcr cytosomc sho\\"s a pattcrn sllggcsti\"c of an carl)' stage of :\IG Lod)' formaiion. l!ppcr cytosome contaillS well developcd :\IG Lod)' with a small satellite structure. Thc upper :\1(; Lody also has a densc core whilc thc lower :\1(; bod)' appcars to bc forming around a less dcnsc core. Arrows indicatc mcmbrancs limiting the cytosomcs. which colllain the :\1(; Lodies. "'.lIuclcus ofhistiocytc. (Uran}"1 acctate and lead citratc stain. X J.t.iOO.)

389

HUMAN PATHOLOGY - VOLUME 4, NUMBER 3

SejJtember 1973

Figure 16. MG body located in intercellular space. Note tendency for apatite crystals arrangement. (Uranyl acetate stain. x 20,000.)

in some mitochondria, the other organelles appeared to be unchanged. No morphologic changes suggesting a transformational relationship between the cytosomes and the mitochondria or the other organelles was seen. MICRo-oRGANls:\ls. No readily recognizable bacterial organism could be seen. Some of the cytosegresomes contained dense material that may at least remotely resemble partially degenerated bacterial components, but these fragments of dense material were not identified among the ~Iichaelis-Guttmann bodies within cytosomes. DISCUSSION

390

Various theories as' to the ongm of Michaelis-Guttmann bodies ha\'e been .proposed o\'er the years since they were

to

have a radial

first described in 1902.28 Mixtures of urine, blood, and bacteria,29'31 specific gram negati\'e bacterial breakdown products,2 degeilCrated erythrocytes,32 mycotic material,33 sarcoid inclusions,34 mineralized mast cells,13 and mineralized cytosegresomes 3. 6 ha\'e all been implicated by numerous in\'estigators. This presentation and three pre\'ious ultrastructural studies did not show e\'idence of ~Iichaclis-Gutt­ mann bodies taking origin from mast cells, erythrocytes, or \'iable micro-organisms.3. 4. 6 ~Iaterial consistent with partially degraded bacterial components was seen, but it is not clear how, or if, this Iliaterial is related to the' specific cytoplasmic inclusions that appear to gi\'e rise to the Michaelis-Guttmann' bodies. Perhaps the products of complete bacterial product degradation contribute to the finely granular homogenous matrix of the cytosome. The possibility of a specific gram negati\'e

MALAKOPLAKIA OF THE SKIN -PRICE ET

AL.

Figure 17. Stereoscopic electron micrographs of clusters of apatite crystals not enclosed within a membrane limited structure, but lying free within cytoplasm. The needle-like configuration of the crystals is appreciated best through a stereo print viewer. (Uranyl acetate stain. x 45,000.)

organism is refuted by the presence of coagulase positi\'e StajJh)1ocOCCllS allrellS in this skin lesion. Those workers who ha\'e postulated the concept of a mineralized cytosegresome ha\'e failed to demonstrate a membrane around the ~Iichaelis-Gutt­ mann bodies or any other proof of transformation from a cytosegresome to a Michaelis-Guttmann body.3.6 This study indicates a likely morphogenetic sequence that results in the ~Iichaelis-Guttmann body. The ~Iichaelis­ Guttmann bodies appear to be caleospherules formed within one type of single membrane limited cytoplasmic inclusion found within the histiocytes. In the earliest stage, prior to mineralization, the cytoplasmic inclusion contains a homogenous, finely granular matrix material and concentric membranous fragments. It resembles the cytosome of the Trump and Ericcson nomenclature,25 the telolysosome described by Gordon et a\.,35 or the homogenous dense bodies,35.36 which ha\'e been referred to as the end stage of phagocy-

tosis. These cytoplasmic inclusions are also probably the ultrastructural manifestation of the small basophilic, periodic acid-Schiff positi\'e cytoplasmic granules seen by light microscopy and suggested by ~IcKeil et a\.4 and later by Smith 1 as the precursor of the Michaelis-Guttmann body. In the next recognizable stage, randomly scattered, small, dense particulate components (as in Figure 8) are seen within the matrix material of the cytosomes. Subsequently the concentrically laminated Michaelis-Guttmann body is de\'eloped, composed of needle shaped hydroxyapatite crystals (Figs. 9, 10, 15, 16)..

The concentric banded pattern of the has been compared to the Liesegang phenomenon seen in gels whereby precipitates form in a pel'iodic ring pattern. 3. 5 Similar laminated calcospherules ha\'e been shown by electron microscopy to constitute the Schaumann bodies surrounding acid fast organisms. 44 They ha\'e also been found in ~Iichaelis-Guttmannbodies

391

HUMAN PATHOLOGY -

VOLU~[E

4, NUMBER 3

SejJtember 1973

Figure 18 Figure 19 Figure 18. Selcctcd arca clcctron micrograph showing ficld samplcd b}" clectron bcam to producc thc clectron diffraction pattcrn of Figurc 19. Accelcrating "oltagc was 100 h. Spccimcn is unstaincd. Figure 19. Sclectcd arca elcctron diffraction pattcrn from a scction of glutaraldchydc fixcd, unstained tissuc using accelerating \"oltagc of 100 h. (i\egath'c print. Thc linc intcnsities \"ar}" slightl}" from original ncgativc from which mcasurements in Tablc I wcrc made; this print is intcndcd mainI}" to show position of lincs.)

392

the extracellular spaces of calcified human aortic valve tissue:17 and in experimentally produced nephrocalcinosis 38 as well as in urinary tract 3 • 4 and gastrointestinal tract malakoplakia. 6 Another interesting feature of this study is the location of mineralization. It takes place within the cell, whereas mineralized materials seen in man and other mammals usually are deposited outside the cell. In higher animals the mineral phase of normal bone formation is an extracellular process,40 and most of the basic pathologic forms of calcification seem to be associated with the extracellular deposition of calcium salts. 41 . 45 Experimental calcification induced in the normal mouse kidney, however, has been shown to be the result of intracellular apatite crystal formation beginning in the mitochondria. 46 Also, Schaumann bodies have been noted to form intracellularly around acid fast organisms in the golden hamster. 44 In single celled organisms, without apparent exception, calcification occurs within the cell, often within single membrane limited vacuoles, which influence the final shape of the calcified object.39

The central problem in calcification is the mechanism by which the salt is laid down. 39 Since most of the research effort to date has been preoccupied with studies of bone, most of the hypotheses available to us have been developed with specific reference to that tissue. A number of theories have been proposed to account for the specific size, shape, and deposition of the particles of calcium phosphate in terms of physical and chemical control of crystal size by an extracellular matrix. 39 • 4o The bone salt is induced, according to these theories, on specific sites for metastable solutions by the stereospecificity of a polysaccharide or protein moiety. The function of a "matrix" in crystal formation has also been emphasized repeatedly in other tissues where calcium phosphate is present in association with a variety of organic matrices, some of which bear no immediate resemblance to those in bone. 39 I n this study the calcium deposits appear to be laid down in the finely granular matrix of the cytosome, which we can only infer indirectly from histochemicaP and our ultrastructural studies to be, at least in part, possibly polysaccharide and lipo-

~IALAKOPLAKIA OF THE SKIN-PRICE ET AL.

TABLE 1.

SELECTED AREA DlffRACTIO:\, DATA FRO~t A SECTJO:\, OF TilE GLUTERALDEIIYDE FIXED ~IALAKOPLAKIA OF TilE SKI:\' CO~II'ARED WITII X-RAY POWDER DIFFRACTJO:\' LITERATURE OF AST~I FILE CARD REFEREXCE 9-432 FOR HYDROXYAPATITE [CALCIU~t HYDROXIDE ORTIIOPIIOSPIIATE, Ca.;(P04 h(OH)]

Selected Area Electron Diffraction of MG Bod}'

(/

d

LillI'

HJdroxyapatite X-raJ Pou·der Diffraclioll Data

/1//"

3.50

~I

3.51 3.44

2 40

2

3.14

\\'

3.17 3.08

12 18

3

2.81

5, B

2.814 2.778 2.720

100 60 60

4

2.26

V\\'

2.296 2.262 2.228

8 20 2

5

1.97

\\'

1.9-13

30

6

1.87

\\'

1.871 1.841 1.806

6 40 20

7

1.76

\\'

1.754 1.722

16 20

8

1.65

VV\\'

1.684

4

9

1.47

\\'

1.503 1.474 10465

10 12 4

10

1.26

V\\'

II

1.17

V\\'

(Plus additional lines not listed on AST~I file card)

12

1.11

V V\\'

*~IG

body:

cytosomes resemble the telol)'sosomes or dense bodies associated ",ith the end stage of lysosomal or autophagoc)'tic activit), may be a clue. Perhaps the pH of this end stage lysosome, as compared to the earlier lysosomal phases, is more conducive to calcification and h)'drox)'apatite formation, in a manner similar to that of the induced formation of h)'droxyapatite crystals in the calcareous corpuscles of cestodes. 42 Further speculation leads one to postulate that this form of calcospherule formation may even represent a specialized intracellular manifestation of calciph),laxis. 43 As indicated in the introduction, malakoplakia has been found not only to involve the mucosa of the lower urinary tracts but has also been reported in the testes, prostate, kidney, stomach, colon, lung, bone, and retroperitoneum. To our knowledge, ho\\'e"er, this is the first example of malakoplakia of the skin. ACKNO\\'LEDGMENT

The authors are grateful to Dr. Sidney S. Pollack, )\Iellon Institute, CarnegieMellon University, for his ,'aluable help in the interpretation of the electron diffraction data. REFERENCES I. Smith. B. H.:

2.

~lichaelis·Gullmann bod)'.

American Society for Testing and ~Iaterials, Philadelphia, Pa. d: Interplanar spacings (hkl) of the crystallallice expressed in Angstrom units. I: ReIati\'e intensities of rings on diffraction pallerns obtained by \'isual inspection (5, strong; ~I, medium; \\', weak; V\\', very weak; VV\\', very, \'cry \\'cak; and B. broad). III.: Relath'e illlensities obtaincd by photometer.

AST~I:

proteinaceous in character. The role, if any, in crystal formation played by the concentric membranes of the cytosomes is not clear; after mature ~Iichaelis-Gutt­ mann bodies were formed, these memb'ranous components were llOt apparent within the cytosomes. The fact that the

3.

4. 5. 6. 7. 8. 9.

~Ialacoplakia of the urinary tract Amer. J. Clin. Path., 43:409, 1965. Terner, J. Y., and Lalles, R.: ~Ialakoplakia of colon and retroperitonenm. Amer. J. CHn. Path., 44:20, 1965. Lambird, P. A .• and Yardley,J. H.: Urinary tract malakoplakia: report of a fatal case with ultra· structural ousen'ations of ~Iichaelis-Gullmann uodies.Johns Hopkins Med.J., /26:1,1970. ~lcKicl, C. F., Jr.. EisellStcin, R., and ~IcDonald, .J. H.: ~Iorphological and microbiological studies in malacoplakia. J. Urol.. 88:236, 1962. Alexander. H. E., and Johnson, P.: Colloid Science. Oxford, Oxford Uni\'crsity Press, 19-19. p. 603. Finlay-Joncs. L. R., Blackwell, J. B., and Papa' dimitriou. J. ~I.: ~Ialakoplakia of the colon. Amer. J. Clin. Path., 50:320, 1968. Haukohl, R. 5., and Chinchinian, II.: ~Ialako· plakia of the testide. Amer. J. Clin. Path., 29:-173.1958. Goldman, R. L: A case of malacoplakia with inwlvemem of the prostate gland. .J. Urol., 93:407, 1965. Hoffman, E., and Garrido, ~I.: ~Ialakoplakia of prostate: report of a case. J. Urol., 92:311, 1964.

393

HUMAN PATHOLOGY - VOLUME 4, NUMBER 3 10. I'urpon, I.. ami Tamayo, R. I'.: ~Ialacoplakia of kidney. J. Urol.. 8-/:231, l!loO. II. Bowcrs, J. H., and Cathay, W. J.: ~Ialakoplakia of kidncy with rcnal failurc. Amer. J. Clin. Path., 55:765, 1971. 12. Gupta. R. K.. Schustcr, R. A., and Christian, \\'. D.: Autopsy findings in a unique case of malacoplakia. Arch. Path.. 93:42. 1972. 13. Yunis, E. J., Estc"ez. J. ~I., Pinzan. G. J.. and ~Ioran. T. J.: ~Ialakoplakia. Discussion of pathogcncsis and repol'l of thrcc cascs including one of fatal gastric and colonic inmh'cmcm. Arch. Path., 83:180,1967. 14. Gonzales-Angulo, A., Corral, E., Garcia-Torres, R., and Quijano, ~I.: Malakoplakia of thc colon. Gastroenterology, 48:383. 1965. 15. DiSih'io, T. V., and Bartlett, E. F.: ~Ialacoplakia of thc colon. Arch. Path., 92: 167, 1971. 16. Rywlin, A. ~I., Ra\'e1, R.• and Hurwitz. A.: ~Ialakoplakia of the colon. Amcr. J. Dig. Dis., 1-1:491. 1969. 17. Ravel, R.: ~Iegalocytic intcrstitial ncphritis. Amcr. J. Clin. Path., 47:781, 1967. 18. ~Iillonig. G.: Aeh'antagcs of a phosphate buffcr for osmium tetroxidc solutions in fixation. J. Appl. Physics, 32:1637,1961. 19. Luft. J. H.: Impro\'cmcnts in epoxy resin cmbedding mcthods. J. Biophrs. Biochcm. Cytol., 9:409, 1961. 20. Ito, S.. and Winchestel', R. J.: Thc fine struclllre of the gastric mucosa in the hat. J. Cell BioI.. 16:541, 1963. 21. Richardson. K. C., jarett, L. and Finkc, E. H.: Embcdding in epoxy rcsins for ultrathin sectioning in clcctron microscopy. Stain Tcchn., 35:313, 1960. 22. Watson, ~1. L: Staining of tissuc scctions for e1cctron microscopy with hcavy metals. J. Bioph}·s. Biochcm. Cytol., -/:475, 1958. 23. Vcnablc. J. B., and Coggcshall, R.: A simplificd lead citratcstain for use in electron microscopy. J. Ccll BioI., 25(2, pI. 1):407. 1965. 24. ·Parsons, D. F.: Thc cxamination of mineral dcposits in pathological tissucs by elcctron diffraction. Int. Rcv. Exper. Path.• 6: I. 1968. 25. Trump, B. F.• and Ericsson, J. L E.: Some ultrastructural and biochcmical consequences of cell il'Uury. III Zwcifach, B. W., Grant. L, and ~IcCluskey, R. T. (Editors): Thc Inflammatory Process. ~ew York. Acadcmic Press. Inc.. 1965. p. 35. 26. Vattcr. A. E.. Zambcrnard, J.. and Pricst, J.: Cell structtlre. III King. D. \\'. (Editor): Ultrastructural Aspects of Diseasc. ~cw York, Hocber Meel. Di\·., Harpcr 8.: Row, HJ66, p. 24. 27. Rcgcr, j. F.• Hutt. ~1. P.• and ~custein, H. n.: Thc fine structure of human hemoglobinuric kidney cells with particular refcrence to hyaline droplcts and iron micellc localization. J. Ultrastrucl. Res., 5:28. 1961. 28. ~Iichaelis, L.. and Guttmann, C.: Ueber cinschlusse 111 B1asemumorcn. Z. Schr. Klin. ~Ied .• 47:208. 1902. 29. Blcisch. V. R.• and Konilson, ~. F.: ~Ialakoplakia of the urinary bladder. Rcport of 4 cases and discnssion of etiology. Arch. Path.• 5-/:388, 1952.

394

SeIdell/bel' 1973

30. Loelc. \\'.: Ein beitrag wr zog: malakoplakic dcr hornblasse. Bcitr. Path. Anat., 48:205, 1910. 31. Rcdcwill, F. H.: Comparison of lcukoplakia. malakoplakia. and cncrusted cystitis: rcport of cascs and a ncw methodoftrcatmcm.J.A.~I.A.• 92:532, H129. 32. Ccdcrquist. L L: ~Ialakoplakia of the urinary tract: a thcory of pathogenesis. At·ch. Path.• 80:495, 1965. 33. Dickson. \\'. E. C.. Gray, A. C. E.• and Kiehl. F.: ~lalacoplakia \'csical: an im'cstigation of ccrtain mycotic infections of the gcnito-urinary tract. Urol. Cman. Rc\·., 31 :611, 1927. 34. French, A. J., and ~Iason,j. T.: ~Ialakoplakiaof urinary bladder and sarcoidosis. J. Urol., 66:229, HJ51. 35. Gordon, G. B.• ~Iillcr, L R.. and Bcnsch. K. G.: Studics on the intraccllular digcstivc proccss in mammalian tissuc culture cclls. J. Ccll BioI.. 25:41, 1965. 36. Hrubin. Z.• and Rechcigl. ~I., jr.: ~Iicrobodics and Relatcd Particles. ~e\\' York. Acadcmic Press, Inc., 1969. 37. Kim, K. ~I., and Huang, S.: Ultrastructural sllldy of calcification of hnman aortic yah·e. Lab. Il1\est., 25:357, 1971. 38. Woodard, J. C.: Amorphologic and biochemical study of nutritional ncphrocalcinosis in fcmalc rats fed scmipurificd dicts. Amcr. J. I'ath., 65:253, 1971. 39. Pautard, F. G. E.: Calcification in uniccllular organisms. III Schraer. H. (Editor): Biological Calcification: Cellular and ~lolcCl1lar Aspects. ~C\\' York. Appleton-Ccntury-Crofts. 1970, p.105. 40. Schiffmann, E.• ~lartin, G. R.• and ~Iiller, E. J.: ~Iatriccs that calcify. III Schracr. H. (Editor): Biological Calcification: Cellular and ~lol­ ecular Aspccts. ~C\\' York. Appleton,CenturyCrofts, 1970. p. 27. 41. Andcrson, W. A. D.: Degencrath'c changes and disturbanccs of mctabolism. 11/ Andcrson. \\'. A. D. (Editor): Pathology. Ed. 6. St. Louis, Thc C. V. ~Iosby Co., 1971, \"01. I, p. 86. 42. \'on Brand, T .• Mcrcado, T. I., ~ylen, ~1. V.. and Scott, D. B.: Observations on function. composition. and structurc of cestodc calcareous corpusclcs. Exp. Parasit., 9:205, I ~J60. 43. Selye, II.: Calciphylaxis. Chicago. Univcrsity of Chicago Press, 1962. 44. Rasmussen, 1'., and Caulfield. J. B.: The ultrastructurc of Schaumann bodics in the golden hamster. Lab. Invcst., 9:330. 'WoO. 45. Freiman, D. G.: Cell injury and ccll death. III Brunson, J. G.• and Gall. E. A. (Editors): Conccpts of Discasc. 1'\e\\' York, The Macmillan Company, 1971. p. 6 I. 46. Caulfield, J. B., and Schrag, P. E.: Elcctron microscopic study of renal calcification. Amer. J. Path., 44:365. 1964. 47. ~euman, W. F.. and ~euman. ~1. W.: Thc Chcmical D}'namics of Bonc Mineral. Chicago, The University of Chicago Prcss, 1958. Pathology Department ~Iagee-\\'omens Hospital Pittsburgh, Pennsylvania 15213 (Dr. Price)