intermediate stage of such a multistep carcinogenic process (8).
Although chemical hepatocancinogens can induce hyperplastic liver lesions within days or a few weeks under optimabconditions, several months or even years are requiredfor eventual cancer production (8). Even then, in spite of theformation of hundreds or possibly thousands of hyperplastic lesions, only a comparatively few cancers ever emerge(6). Fewer still are the hepatocellubam carcinomas that prog
messto aggressive metastasizing lesions. What conditionsaffect the degree and the rate at which precursor cellsacquire their ultimate fully malignant potential?
A variety of factors could theoretically intercede to accel
erate,retard,orotherwisemodulatethecourseofcarcinogenesis following the initial interaction of a carcinogen withthe target cell. Of these factors, perhaps none is more apt toexert a crucial modulating influence than blood supply. Byregulating influx of exogenous and endogenous promotingagents, hormones, growth factors, toxic substances, envimonmental carcinogens, and nutrients, alterations in bloodflow can reasonably be expected to affect the biological fateof preneoplastic tissue.
This supposition gains some indirect support from thework of Folkrnan et al. (9) whose findings demonstrate theprofound biological effect that neovasculanization exerts onfrankly malignant tissue. Capillary ingrowth elicited by atumor product, “tumorangiogenesis factor,―appears to bean obligatory step for actuation of malignant properties inthe late stages of development of some tumors (10). Similanly, alterations in the blood supply to putative preneoplastic lesions and primary tumors may have significant biobogical consequences in earlier stages of cancinogenesis.
Radioactive microspheres have been used experimentallyto study the circulation in diverse pathological processeswith attendant alterations in blood flow (1, 3, 12, 17). Whenintroduced into the vascular system, spheres of the appropmiate size will have negligible recirculation capacity andrapidly lodge in the micnovasculatume. At appropriate dosesthey have no observable effect on the gross physiology ofthe animal. Errors and limitations of their use have beenreported (5). The application of y-Iabeled microspheres forthe quantitative measurement of arterial and portal flow tohyperplastic nodules and hepatomas in comparison withthe surrounding liver is the subject of this paper.
MATERIALS AND METHODS
Animals and Treatment. Male Fischer 344 rats (150 to 200mg; Charles River Breeding Laboratories, Wilmington
SUMMARY
Intravascular injection of radionuclide-labebed microspheres was used to compare the blood supply to diethylnitrosarnine-induced hyperplastic liver nodules and hepatomas with the blood supply to the surrounding, histologicallynormal liven. Microspheres injected into the heart on portalvein lodged in the organs of control and diethylnitmosaminetreated rats providing a quantitative index of blood supplyto the microvasculan bed. The blood supply is expressed aspercentage of cardiac output (arterial) or cpm (portal) penorgan, lobe, g tissue, etc.
The fraction of the cardiac output received by lung, kidneys, spleen, and liver was similar in control and carcinogen-treated animals. The arterial blood supply of 23 nodulesand hepatornas was variable [1.17 ±0.22% (S.E.) cardiacoutput per g, fixed weight], but it was similar to the arterialsupply to the surrounding tissue (1.12 ±0.21% cardiacoutput per g, fixed weight). In contrast the portal bloodsupply to 25 selected lesions was 39 ±6% that of thesurrounding liver tissue. Themewas no apparent relationship between blood supply and lesion size or histologicalappearance.
While only 0.13 ±0.04% of the microsphemes injected viathe portal system were recovered in the lungs of controlrats, approximately 100 times this number bypassed or escaped the liver containing nodules and hepatomas andlodged in the lungs.
Such alterations in blood flow could contribute to biobogicabdiversification of hepatic lesions in successive stages ofcancer evolution and could facilitate metastasis from theliver.
INTRODUCTION
The concept that cancer may result from a multistepprocess of cellular evolution rather than from a single immutable event is gaining prominence in oncology (7, 11).Mounting evidence suggests that chemically induced hyperplastic liven nodules may be one example of tissue at an
I Supported by grants from the National Cancer Institute of Canada, The
Connaught Fund of the University of Toronto, and The Medical ResearchCouncil of Canada (MA-1056).
2 Supported by a Training Grant (DE-2268) from the National Institute of
Dental Research to Temple University.3 To whom requests for reprints should be addressed, at Banting Institute,
100 College Street, Toronto, Ontario, Canada M5G 1L5.Received November 15, 1976; accepted March 1, 1977
1686 CANCERRESEARCHVOL. 37
Comparison of the Blood Supply to Diethylnitrosamine-inducedHyperplastic Nodules and Hepatomas and to the SurroundingLiver1
Dennis B. SoIt,2 John B. Hay, and Emmanuel Farber3
Blood Supply in DENA-induced Liver Nodules and Hepatomas
Mass.)were maintainedon a high-protein(26%)basaldiet(Bioserv, Inc., Fmenchtown, N. J.). For induction of hyperplastic liver nodules and hepatocellular carcinomas, DENA4in 0.9% NaCI solution was injected i.p. at a dose of 200 mg/kg/injection. Animals received either 2 on 3 such injectionsspaced at 2-week intervals. Discrete, grossly visible liverlesions, both nodules and cancer, were reproducibly induced, with the intervening liver generally normal in histological appearance (Figs. 1 and 2). Control animals were allgiven 3 similarly spaced injections of 0.9% NaCI solution.This study was performed in the 9th month after the initialcarcinogen administration. At this time the surviving (90%)carcinogen-treated matswere 230 to 320 g. Control animalswere 310 to380g.
Labeled Microspheres. Tracer Sephadex (15 ±3 @rn)(Pharmacia Fine Chemicals, Inc., Piscataway, N. J.) waslabeled with 141Ceaccording to the manufacturer's specifications. The specific activity was 0.5 to 4 cpm/spheme underour counting conditions. The total administered dose wasvaried. In studies measuring the blood supply to individualliver lesions, the administered dose was approximately 8.2x [email protected] 10@cpm and 4.5x [email protected] 10@cpm forarterial and portal rneasunments, respectively.
Arterial Blood Flow Measurements. Light anesthesia wasinduced with 0.25 to 0.30 ml of sodium pentobambital (60mg/mI i.p.), The might carotid artery was dissected free ofsurrounding tissue, and 2 loose 4-0 silk ligatures wereplaced around the vessel approximately 1 cm apart. A PE-60polyethylene catheter (Clay-Adams, Parsippany, N. J.) contaming a solution of 0.9% NaCI and 5 IU of hepanin per ml(NaCI:hepaminsolution) was pulled out to a narrow diameterto facilitate entry into the vessel. The vessel was pierced,and the catheter was introduced and advanced approximately 3.5 cm into the heart. A small amount of blood wasaspirated to ensure patency of the catheter.
Tracer Sephadex suspended in 0.5 ml of NaCI:hepaminsolution was rapidly injected through the catheter. An add itional 1 ml of solution was rinsed through the syringe towash residual microsphenes into the heart. The animal wasthen sacrificed with an i.c. injection of sodium pentobanbitab. Individual liver lobes and other organs were removed,weighed, placed in vials, and counted ifl a IntertechniqueModel CG3Ogamma spectrometer. Syringe counts obtainedbefore and after infusion provided the net amount of radioactivity administered, i.e., cardiac output in cpm.
Portal Blood Flow Measurements. Following sodiumpentobarbital anesthesia a lapanotomy was performed toexpose the portal vascular tree. Labeled tracer Sephadexwas slowly injected through a 25-gauge needle retrogradeinto the distal aspect of the superior mesentemic vein. Theinjection volume was 0.6 to 0.8 ml of Sephadex inNaCI:hepanin solution tinted with Evans blue. Trial infusionsdemonstrated a uniform distribution of dye in the liver lobesof control animals when injections were performed in aretrograde manner to minimize streaming in the portal yessel. Animals were sacrificed by exsanguination via the abdominab aorta. Liver lobes were individually excised,weighed, and counted in a gamma spectrometer.
4 The abbreviations used are: DENA, diethylnitrosamine; ic. , intracardiac.
Measurements of Blood Flow to Lesions and SurroundIng Liver. Analysisof blood flow (microsphenedistribution)to lesions and surrounding liver was performed followingfixation of each liver in Carnoy's fixative (ethanol:chboroforrn:acetic acid, 6:3:1). Gross serial sections approximately1 mm thick were made of each lobe to include selectedlesions and the surrounding liver. All lesions were roughlyspherical and usually less than 1 cm in diameter. Followingfixation they were light gray to tan and clearly demarcatedfrom the darker tan surrounding liven. A section of eachlesion including the surrounding liver was processed forhistological examination. From the remaining serial sections, the coin-shaped portions of each lesion were dissected free of surrounding liver, individually pooled, andimmediately placed in airtight preweighed counting vials.The portions of fixed liver surrounding each lesion werecollectively pooled for measurement of “surrounding―blood flow in each liver. After being weighed, the sampleswere counted and blood flow was expressed as percentageof cardiac output (arterial) or cprn (portal) per 100 mg offixed tissue.
Statistical Methods. When appropriate, the significanceof the difference between arithmetic means was determinedusing an independent Student t test, and the p values areindicated. Unless otherwise designated the other valuesshown represent the mean ±S.E.
RESULTS
Distribution of Arterial and Portal Blood Flow amongIndividual Liver Lobes. The validity of the technique used inthis study depended upon adequate mixing of the microspheres in the blood and their widespread and uniformdistribution in the liver tissue. As a test of this, microspheres were introduced into either the arterial or portalsystem of control animals and distribution of radioactivitywas measured in relation to the weight of individual liverlobes. Chart 1 shows that both methods of administrationresulted in uniform distribution of labeled particles amongthe lobes.
As an additional check on the uniformity of microsphenedistribution, a liven was fixed after each method of sphereadministration, random fragments from each lobe wereweighed, and the radioactivity was determined. Followingarterial administration 17 fragments (257 ±25 mg) from oneliven trapped 2517 ±263 (S.E., 10% of the mean) cpm/100mg. For the other liver given spheres via the portal system,18 random fragments (189 ±22 mg) had 1810 ±150 (S.E.,8% of the mean) cprn/100 mg. The low standard errors in
each case are an additional indication that sphere distnibution is reasonably uniform within a normal liver using thistechnique.
Organ Distribution of Cardiac Output In Normal andDENA-treated Rats. The percentage of the cardiac outputreceived by each of 4 different rat organs is shown in Chart2. In control animals the values for liver, kidneys, lung, and
spleen were 6.0 ±1.2, 17.9 ±2.0, 2.8 ±0.9, and 0.9 ±0.1%,respectively. This distribution is very similar to that previously reported using the microsphere technique in the rat(16). In DENA-treated rats the fraction of the cardiac output
only 39 ±6% as much portal blood (i.e., spheres, cpm/g,etc.) as the surrounding liver. Only 1 of the 25 lesionsappeared to receive more portal blood than its surrounding(L/S = 1.25). For example, L/S x 100% for 8 lesions inAnimal P4 ranged from 1 to 125% and averaged 28 ±15% (L= 115 ± 62 cpm/100 mg; S = 421 cpm/100 rng).
Comparison of Lesion Histology, Size, and Blood Supply. A histopathological study of 48 lesions failed to elucidate any relationship between their histological pattern andtheir portal or arterial blood supply. The lesions includedtypical hypemplastic nodules (Fig. 1), unequivocal hepatocellular carcinomas (Fig. 2), and a spectrum of intermediatelesions. No lung metastasis, anaplastic carcinomas, on extensive necrosis were encountered. Each lesion was diagnosed independently by 3 pathologists and graded from 1(nodule) to 10 (hepatoma), with respect to pleomorphism,mitotic activity, nuclean:cytoplasmic ratio , basophilia, nucleolan prominence, and several other criteria. No convincing association could be drawn with lesion blood supply (L/5) and any of the histological parameters considered.
There was also no apparent correlation between lesionsize and portal or arterial blood supply (Chart 3). Thus,theme is no suggestion that blood flow per g of tissue increased (or decreased) as the lesions grew.
7-
a)
0a
20
15
10-@
LIVER KIDNEYS LUNG SPLEEN
I0
rT,0=
Ea
L@J>
-J0U
LOBE
Chart 1. Distribution of label among individual liver lobes following arterial or portal administration of labeled microspheres to normal rats. The totalcpm for each lobe are expressed as percentage of whole-liver cpm andcompared to the proportional liver weight of each lobe. Bars, mean of 4(arterial) or 5 (portal) rats ±S.E.
received by kidneys, lung, and spleen was similar to controls. An apparent elevation in the percentage of cardiacoutput received by the liver of carcinogen-treated mats(treated 10.0 ±1.3% cardiac output versus control 6.0 ±1 .2% cardiac output) may not be significant (0.10 > p >
0.05).Comparison of Arterial and Portal Blood Flow to Lesions
and Surrounding Liver. Table 1 summarizes the arterialblood flow measurements from DENA-treated rat livers. Theaverage blood flow (5) to the lesion-free surrounding liverwas 1.12 ±0.21% of the cardiac output per g of fixed liver.Although there was a 4-fold variation in S from 0.42 to1.65%, the values may not differ significantly (0.10 > p >0.05) from those obtained from 4 control livers (0.40, 0.53,0.94, and 0.38%).
The average arterial blood flow (L) to 23 fixed lesions was1.17 ±0.22% cardiac output pen g. While this average valueis similar to that obtained for surrounding liver (1.12 ±0.21%), the values for individual lesions (not shown) variedwithin a given liven and ranged from 0.03 to 4.39% cardiacoutput per g among the 6 animals.
A comparison of arterial blood flow to 23 lesions and totheir surrounding liver revealed no consistent relationshipbetween the 2 (Chart 3). L/S x 100% for individual lesions(not shown) in the 6 mat livers varied from 2 to 294% andaveraged 112 ±15%.
In contrast, measurements of the portal blood flow distnibution in 5 treated nat livers reveal a striking difference inthe portal L/S ratio (Table 2; Chart 3). The lesions received
:D00
0
0
Chart 2. Organ distribution of cardiac output received by 4 organs incontrol and DENA (DEN)-treated rats. Following ic. administration of labeledmicrospheres, the label lodged in each organ was expressed as percentageof cardiacoutput (i.e., percentageof thetotal administereddose).Thevaluesfor 4 control rats (% ±S.E.)were: liver, 6.0 ±1.2; kidneys, 17.9 ±2.0; lungs,2.8 ±0.9; and spleen,0.9 ±0.1. Valuesfor 6 DENA-treatedrats (% ±S.E.)were: liver, 10.0 ±1.3; kidneys, 18.4 ±3.6; lung, 2.7 ±0.2; and spleen, 1.1 ±0.2.
Blood Supply in DENA-induced Liver Nodules and Hepatomas
Liver to Lung Bypass of Portal Microspheres in DENAtreated Rats. Over 99% of the labeled microspheres injected into the portal venous system of 5 control matsappeared to lodge in the liven. Only 0.13 ±0.04% (range, 0.04to 0.25%) of the administered cprn (i.e., spheres) passedthrough the liver to lodge in the lung. In contrast whenmadionuclide-labebed micnosphemes were injected into theportal venous system of 10 DENA-treated rats with nodulesand hepatomas, 14 ±4% (range, 5 to 38%) of the totaladministered cpm were detected in the lung.
DISCUSSION
A few reported quantitative studies comparing the portal
on arterial blood flow to liven tumors and the surroundingIiver(3, 4, 17) have dealt primarily with metastatic or transplanted carcinomas. Comparable studies on the blood sup
300
200
AA
A
A
A
C
100 . AA e
AC A
A
A e
CC Ac
A e CeCe
CCC
C AA :@
CCC1C@ A Im
Table 1
100 200 300 400
LESION(fixed) WEIGHT-mg
Chart 3. Individual lesion portal (•)or arterial (Li) blood flow versus lesionsize. Following ic. or intraportal administration of radiolabeled microspheres, the blood flow to individual lesions was expressed relative to thesurrounding liver by:
L (i.e. , cpm/g lesion)x 100%
S (i.e., cpm/g surroundings)
Thesevaluesforeachlesionareshownplottedagainsttheweightof the fixedlesion. An additional lesion is not depicted [@(arterial): L/5 x 100% = 173%;weight, 1128mgj.
ply to primary hepatomas and presumptive preneoplasticlesions have been largely neglected, possibly because ofthe experimental difficulty in developing such lesions in theabsence of general distortion in liver architecture and hemodynamics. The carcinogenic regimen used in this studyminimizes such distortion while promoting the formation ofdiscrete hyperplastic nodules and hepatomas.
Blanchand et a!. (3) used labeled micnosphemes to measure the blood flow to V2 carcinoma implants and surrounding liver. After portal infusion the concentration of label inV2 tumors was 23% that of the surrounding liver. Thisapparent decrease approaches 39 ±6% measured for portalflow to DENA-induced hyperplastic nodules and primaryhepatomas. However, when micnospheres were infused intothe hepatic artery, themewas a variable but consistent elevation of label in V2 implants from 1.2 to 10.2 times that of thesurrounding liver. This result differs from the arterial measurements of DENA-induced lesions in which the concentration of labeled microspheres, although variable, averaged1.1 times that of surrounding liven. These combined resultssuggest that the more aggressive V2 carcinoma may haveacquired a capacity for arterial neovasculanization not yet
a P1 and P2 received a total i.p.DENA dose of 400 mg/kg DENA.
P3, P4, and PS received a total i.p. DENA dose of 600 mg/kg.b Mean ± S.E.
new capillaries supplying the developing lesions with nutnient-mich blood partially detoxified by the surrounding liven.While obviously thenoetical, this scheme is in agreementwith our present limited knowledge of the development ofliver cancer in man and animals.
ACKNOWLEDGMENTS
We wish to express thanks to Dr. Alan Medline and Dr. Ross Cameron forassistance in evaluating histological slides and to Esther Deak, StanSchockey, and John Hendricks for excellent technical assistance.
REFERENCES
1. Ackerman, N. B., Lien, W. M., Kondi, E. S., Silverman, N. A. The BloodSupply of Experimental Liver Metastasis. I. The Distribution of HepaticArtery and Portal Vein Blood to “Small―and Large―Tumors. Surgery,66: 1067-1072,1969.
2. Bierman, H. R., Byron, R. L., Jr., Kelley, K. H., and Grady, A. Studies onthe Blood Supply of Tumors in Man. III. Vascular Patterns of the Liver byHepatic Arteriography in vivo. J. NatI. Cancer Inst., 12: 107-131 , 1951.
3. Blanchard, R. J. W., Grotenhuis, I., LaFave, J. W., Perry, J. F., Jr. BloodSupply to Hepatic V2 Carcinoma Implants as Measured by RadioactiveMicrospheres. Proc. Soc. Exptl. Biol. Med., 118: 465-468, 1965.
4. Breedis, C., and Young, G. The Blood Supply of Neoplasms in the Liver.Am. J. Pathol., 30: 969-985, 1954.
5. Buckberg, G. D., Luck, J. C., Payne, D. B., Hoffman, J. I. E., Archie, J.P., and Fixler, D. E. Some Sources of Error in Measuring Regional BloodFlow with Radioactive Microspheres. J. AppI. Physiol., 31: 598—604,1971.
6. Epstein, S. M., Ito, N., Merkow, L. P., and Farber, E. The CellularAnalysis of Liver Carcinogenesis: The Induction of Large HyperplasticNodulesin the Liverwith 2-Fluorenylacetamideor Ethionineand SomeAspects of Their Morphology and Glycogen Metabolism. Cancer Res.,27: 1702-1712,1967.
7. Farber, E. Carcinogenesis—Cellular Evolution as a Unifying Thread:Presidential Address. Cancer Res., 33: 2537-2550, 1973.
8. Farber, E. Hyperplastic Liver Nodules. Methods Cancer Res., 1: 345—375,1973.
9. Folkman, J. Tumor Angiogenesis: A Possible Control Point in TumorGrowth. Ann. Internal Med., 82: 96-100, 1975.
10. Folkman, J. , Merler, E., Abernathy, C., and Williams, G. Isolation of aTumor Factor Responsible for Angiogenesis. J. Exptl. Med., 133: 275-288, 1971.
11. Foulds, L. Neoplastic Development, Vol. 1, Chap. 3, pp. 41-89. NewYork, Academic Press, Inc., 1969.
12. Hay, J. B., Johnston, M. G., Hobbs, B. B., and Movat, H. Z. The Use ofRadioactive Microspheres to Quantitate Hyperemia in Dermal Inflammatory Sites. Proc. Soc. Exp. Biol. Med., 150: 641-644, 1975.
13. Healey, J. E., Jr. Vascular Patterns in Human Metastatic Liver Tumors.Surg. Gynecol. Obstet., 120: 1187-1193, 1965.
14. Krishna Murthy, A. S. Vascular Pattern in the Induced Primary Carcinomaof the Liverof Rats.Brit. J. Exptl. Pathol.,40: 25-32,1959.
15. Mann, J. D., Wakim, K. G., and Baggerstoss, A. H. Alterations in theVasculature of the Diseased Liver. Gastroenterology, 25: 540-556, 1953.
16. Sapirstein, L. A., Sapirstein, E. H., and Bredemyer, A. Effect of Hemorrhage on the Cardiac Output and Its Distribution in the Rat. CirculationRes., 8: 135-148, 1960.
17. Sugahara, K., Mitani, S., Shirakura, T., Kawano, N., Kashii, A., andIshikawa, K. Hepatic Dearterialization in 3-Methyl-4-Dimethylaminoazobenzene-induced Hepatocellular Carcinoma with Special Reference toCirculatory Dynamics and Mitochondrial Functions. Gastroenterology,68: 1278—1283,1975.
expressed by the majority of DENA-induced nodules andprimary tumors in this series.
There was no obvious correspondence between the histological appearance of individual lesions and their bloodsupply. While the lesions included typical hypemplastic nodules and classical trabeculan carcinomas, no metastasizingor anaplastic tumors were encountered. In addition, the sizemangeof the fixed lesions was rather narrow (90% < 300mg). On morphological grounds then, the lesions may all beconsidered to baytoward the benign end of the neoplasticspectrum. Consistent with this interpretation is the appanent inability of the lesions to evoke neovasculanization accompanied by a large increase in arterial blood flow.
Of the spheres injected into the portal venous system 14± 4% bypassed the liver with hypenplastic nodules and
hepatomas and lodged in the lungs. This bypass may haveresulted from anastomosis of the portal and systemic yenous system within the liven lesions. The presence of barge(microscopic) blood-filled channels within some of the besions studied adds support to this possibility. In additionseveral of the livers contained cavernous vascular sinuses,often grossly visible on the capsular and Cut surface, whichsuggests another possible explanation for the bypass phenomenon. The presence in DENA-treated rats of hepaticthoroughfares from the portal to the systemic venous system could provide an ideal route for liver to lung metastasisin subsequent stages of tumor development. There was no
evidence (e.g., enlarged spleen cirrhosis, ascites, etc.) tosuggest the alternate possibility of extnahepatic bypass viacollateral vessels, as seen for example with some types ofhuman portal hypertension.
The results of our study and previous studies (2—4,13—15,17) indicate that all types of neoplastic lesions in the liver,
including hypemplastic nodules , hepatomas , and metastasisin man and animals, can be associated with a relative decrease in portal blood flow when the lesions are comparedto the surrounding liven. This alteration in blood flow couldhave important biological consequences with respect totumor evolution.
Once formed, “pneneopbastic―or early malignant lesionswould be partially protected from bacteria or other toxins(e.g. , drugs) arriving from the gastrointestinal tract, whilethe surrounding liven would be left to perform the bulk ofdetoxification and the usual gamut of other metabolic functions. Gradually, oven a period of several months or years,this physiological imbalance, augmented by periodic influxof environmental (or experimentally administered) cancinogens, could inhibit growth of the surrounding liver whileexerting a promoting influence on the lesions. Furthergrowth imbalance could possibly result from ingrowth of
Fig. 1. Hyperplastic liver nodule (top and right of center). Portal blood flow appeared reduced in this nodule (LIS x 100% = 53%) using the microspheretechnique. The surrounding liver tissue has a 1-cell-thick plate pattern resembling that of normal liver. H & E, x 250.
Fig. 2. Hepatocellular carcinoma (top and right ofcenter) from same liver as the nodule shown in Fig. 1 . The tumor has a trabecular pattern with numerouslarge vascular channels. Mitoses and nuclear pleomorphism are evident. The histological pattern of the surrounding liver tissue is generally similar to that ofnormal liver, with 1-cell-thick plates predominating. The microsphere technique indicated a greatly diminished portal blood flow (LIS x 100% = 1%) in thislesion.H & E, x 250.
1977;37:1686-1691. Cancer Res Dennis B. Solt, John B. Hay and Emmanuel Farber LiverHyperplastic Nodules and Hepatomas and to the Surrounding Comparison of the Blood Supply to Diethylnitrosamine-induced
![Geomantia Nova [1686]](https://static.documents.pub/doc/80x56/56d6bdd21a28ab30168f78d2/geomantia-nova-1686.jpg)
![Semiphoras vnd Schemhamphoras Salomonis Regis [1686]](https://static.documents.pub/doc/80x56/56d6bf301a28ab3016953bee/semiphoras-vnd-schemhamphoras-salomonis-regis-1686.jpg)
