Biological activities of phthalocyanines. XVII histopathologic evidence for different mechanisms of EMT-6 tumor necrosis induced by photodynamic therapy with disulfonated aluminum phthalocyanine or photofrin.

The necrosis of EMT-6 mammary murine tumors induced by photodynamic therapy (PDT) with Photofrin (PII) or disulfonated aluminum phthalocyanine (AlPcS2) was studied. Attention was given to the spontaneous evolution of angiogenesis and necrosis of such tumors in order to determine the most appropriate moment for treatment. On day 6 after tumor cell inoculation, mice were injected with photosensitizer followed by exposure to red light 24 h later, at which time optimal dye concentrations were reached in the tumor. Animals were sacrificed 3 h or 24 h after illumination and tissues were prepared for histology. Prominent cytopathic alterations were already observed at 3 h and there was massive necrosis at 24 h. In the case of PII vascular damage, congestion and hemorrhage were already present at 3 h and these changes account for the subsequent tumor necrosis through hemorrhagic infarction. With AlPcS2 these early vascular alterations were much less evident and only focal at 24 h, suggesting that AlPcS2-PDT mediated tumor necrosis involves to a large extent direct tumor cell damage.

Innocuity of agents for radiocontrast enhancement of urinary tract stones.

OBJECTIVES: Mineral kidney stones are frequently difficult to detect due to their radiotranslucency. We have recently developed a method that enhances the visibility of such stones by retrograde infusions of certain heavy metal salt solutions such as cesium or lanthanide gadolinium. This report describes toxicologic studies carried out on the use of those contrast agents to introduce this technique eventually into clinical trials. METHODS: Systemic absorption was assessed in dogs through infusion of radioactive contrast agent into the renal pelvis with or without ureteral obstruction. Radioactivity in urine and blood was monitored. Local toxicity was studied in animals infused with the contrast agent at intervals up to 4 weeks. RESULTS: Reabsorption studies under high intrapelvic pressures (70 cm H2O or higher), demonstrated reabsorption of cesium. However, at normal intrapelvic pressures, only a moderate reabsorption of cesium was observed. No gadolinium reabsorption was detected even at high intrapelvic pressures. Histopathologic studies showed no major urothelial lesions but only a transient inflammatory reaction that was undetectable 4 weeks following exposure to gadolinium salts. CONCLUSIONS: Gadolinium salt solutions are good positive radiocontrast agents for mineral kidney stones without having serious toxic effects or systemic reabsorption.

Meniscal repair by synovial flap transfer. Healing of the avascular zone in rabbits.

We studied repair of a longitudinal incision of the right medial meniscus in 44 rabbits after the transfer of a pedunculated synovial flap, without immobilization of the knee. The left medial meniscus was used as the control, after creating the same lesion without synovial flap. Healing was analyzed by histologic studies, including India ink perfusion after 8, 12, 24, and 48 weeks. In three quarters of the cases, the meniscus showed healing with vascularization of an originally avascular zone.

An animal model of Kaposi's sarcoma. I. Immune status of CD1 mice undergoing dimethyl hydrazine treatment to induce angiosarcomas and other malignancies.

Dimethylhydrazine (DMH) induces colonic cancer and angiosarcomas in mice. In order to determine pertinence of mouse angiosarcoma as a model to AIDS associated Kaposi's sarcoma (KS), we investigated if immune dysfunction occurred during tumor development by DMH. Outbred CD1 male mice received once weekly DMH a 20 mg/kg body weight dose s.c. for 33 weeks. Every two weeks initially and then every week groups of DMH-treated and control animals were sacrificed to determine a) peripheral blood and splenic T cell subset ratio b) 4-day plaque forming cell (PFC) response to i.p. sheep red blood cells (SRBC) and c) mitogenic response of spleen cells to Concanavalin A (Con A) and lipopolysaccharide (LPS). No change in T helper/T suppressor + cytotoxic T cell (Th/Tsupp. + CTL) and mitogenic response to spleen cells to Con A was noted whereas PFC response of animals to SRBC and mitogenic response of spleen cells to LPS decreased. These data suggest that either infection with T cell depleting virus such as LP:BM5 or immunosuppressive drugs affecting T cell function, such as steroids may be required to bring the immune status of DMH treated animals closer to that of AIDS associated KS bearing human subjects.

An animal model of Kaposi's sarcoma. II. Pathogenesis of dimethyl hydrazine induced angiosarcoma and colorectal cancer in three mouse strains.

CDI, C57B1/6 and DBA2 mice were subjected to dimethyl hydrazine (DMH) carcinogenesis to determine incidences of various types of tumors developing internally in the three strains. The animals were also screened for skin angiosarcomas. Skin lesions histologically similar to cutaneous AIDS associated Kaposi's sarcoma were observed in CD1 mice. Angiosarcomas predominated over colorectal tumors in C57Bl/6 and DBA2 mice, whereas the reverse was true for CD1 mice. The visceral angiosarcomas had both histological similarities and differences with visceral AIDS associated Kaposi's sarcoma.

Radio-contrast enhancement of urinary tract stones.

Most urologists treating stone disease with any method (ESWL, PCL, URS) have encountered problems of poor stone visualization with fluoroscopy. This difficulty to localize urinary tract (UT) stones or fragments may result in incomplete stone extraction, prolonged surgery and increased risk of recurrence and post-operative complications. We have sought and found means to increase the radioopacity of mineral UT stones by a simple pre-operative perfusion technique. The capacity of radioopacification has first been demonstrated in in vitro incubations of fragments of human mineral stones with aqueous solutions of barium, of the lanthanides and of the two natural actinides. Most of the incubations led to considerable radio-contrast enhancement and heavy metal incorporation, measured by X-ray fluorescence analysis. Dogs with implanted human stone fragments were used as an in vivo model. The UT were perfused through a retrograde pyelic catheter with heavy metal salts solutions, the ensuing radioopacification of the implanted UT-stones was estimated by abdominal radiographies and the metal incorporation was measured on the retrieved stones. Considerable radioopacity enhancement together with heavy metal incorporation was observed for the following elements: Sr, Ba and the lanthanides Gd and Yb. The pathological evaluation of the urothelial linings from animals treated with lanthanide salt showed no toxic effects.

Effect of maltose tetrapalmitate on tumor vascularization and tumor growth.

C3HBA mammary tumor (H-2k) was implanted s.c. by trocar in MTP responder C3H/HeN (H-2k) and non-responder C3H/HeJ (H-2k) female mice. One half of the animals received MTP i.p. The size of the tumor was measured everyday. Tumor growth was slightly slower in the C3H/HeJ than in the C3H/HeN. MTP treatment was effective only for the tumor implanted in the C3H/HeN. Microscopic and microscopic visualization of the tumor 1, 2, 3, and 15 days after tumor implantation and MTP treatment revealed poor vascular development in the MTP-treated C3H/HeN compared to untreated controls at days 2 and 3.

Evaluation of cortisone-heparin and cortisone-maltose tetrapalmitate therapies against rodent tumors. I. Biological studies.

The antitumor activity of either cortisone-heparin or cortisone-maltose tetrapalmitate combination or both was tested against two animal tumor models. The first model was orthotopically implanted bladder tumor established in syngeneic Fisher 344 rats. Shrinkage and growth arrest of the tumors were induced by cortisone and amplified by its combination with either heparin or maltose tetrapalmitate (MTP). The second model was trocar implanted C3HBA mammary tumor piece s.c. in syngeneic LPS and MTP responder C3H/HeN and non responder C3H/HeJ mice. The tumor was sensitive to growth inhibition by cortisone-MTP in the C3H/HeN but not by cortisone alone or cortisone-heparin. Tumor implanted in C3H/HeJ was much less sensitive to cortisone-MTP. Cortisone could be replaced by 17-alpha-hydroxyprogesterone, but not by cortexolone.

Evaluation of cortisone-heparin and cortisone-maltose tetrapalmitate therapies against rodent tumors. II. Pathological studies.

Pathological studies were undertaken in three tumor-host models which were subjected to cortisone based treatments. The first model was Fisher 344 rats with established orthotopically implanted syngeneic bladder tumor. Cortisone-herapin and cortisone-maltose tetrapalmitate (MTP) treatments induced focal areas of tumor necrosis and necrobiosis, whereas cortisone alone caused necrobiosis. The second model was C3HBA mammary tumor fragments implanted subcutaneously in syngeneic MTP responder C3H/HeN and MTP non-responder C3H/HeJ female mice. Only cortisone-MTP treatment led to an absence of capillary extension from surrounding blood vessels into the scant tumor stroma. The third model, ethyl carbamate induced primary lung cancer in AJ mice, was tested only with cortisone-herapin combination. The treatment caused central zones of necrosis.

The reliability of routine pathologic diagnosis of colorectal adenocarcinoma.

The diagnosis of colorectal adenocarcinoma is generally considered as being reliable. However, the reproducibility of the classification of specific histologic patterns and the interrater agreement on the gradings have not been firmly established. A panel of three independent expert pathologists reviewed histologic sections from 128 patients selected among 1848 with colorectal cancer, diagnosed in 11 hospitals of the same region. The panel agreed with 92.6% of the original diagnoses of colorectal adenocarcinoma. As for agreement between panel members, the kappa value was 0.78 for the diagnosis of adenocarcinoma and 0.62 for confirmation of colorectal origin. The intraclass correlation coefficient for tumor differentiation features was 0.75. The proportion of villous and adenomatous components also generated good agreement. However, the grading of mucin secretion showed poor agreement (intraclass correlation coefficient = 0.44). Results confirm the reliability of routine pathological diagnosis and also demonstrate the reproducibility of basic diagnostic categories and pathognomonic features. Thus, to obtain reliable information from medical records for epidemiologic and clinical studies, data should be limited to well-defined diagnostic and histopathologic categories.

Prophylactic antiangiogenic tumor treatment.

Complementary antitumor treatments are required to increase the cure rate achieved by surgery and/or radiotherapy by avoiding future recurrences and metastases. The growth of most solid tumors, particularly carcinomas, depends upon the simultaneous development of internal tumor vasculature to allow the proliferation of tumor cells. Inhibition of tumor vascularization is an indirect means of limiting tumor expansion. Daily administration of cortisone and maltose tetrapalmitate (MTP) abolished growth of implanted syngeneic C3HBA mammary tumor. Gross and macroscopic examination of these tumors revealed that tumor growth was prevented. Histological examination demonstrated lack of vascularization within the neoplastic tissue. We believe that this combination in an appropriate form could provide a prophylactic treatment regimen after conventional antitumor treatments in humans.

Antitumor efficacies of maltose tetrapalmitate immunotherapy alone and in combinations with radiotherapy and with cyclophosphamide chemotherapy against dimethylhydrazine induced colon and anal cancers in CDI mice.

Treatment of human colonic cancer in early stages when the process is still limited to the colonic wall is primarily surgery. We wished to see if maltose tetrapalmitate (MTP) immunotherapy alone or in combination with radiotherapy (R) and cyclophosphamide (C) chemotherapy would be effective against primary colon cancer in a fashion similar to that reported by us for primary liver cancer (Anticancer Research 6: 245-250, 1986). One hundred female CD1 mice were subjected to dimethylhydrazine (DMH) treatment once a week for 26 weeks, a period one week before which, colon cancer was histologically documented in each animal of a group that was sacrificed. Surprisingly, many of the animals harboured early anal cancer as well. At 28 weeks, 85 of the available animals were divided into 6 groups that received: Gr. 1, no treatment; Gr. 2, MTP alone (M); Gr. 3, radiotherapy alone (R); Gr. 4, cyclosphophamide alone (C); Gr. 5, R + C; Gr. 6, M + R + C. Criteria of treatment efficacy were: number, size and staging of colorectal tumors and the incidence and the size of anal tumors at death. Mean survival time was also determined although it remained a questionable criterium since most animals died due to complication (hepatic toxicity, pyelonephritis, thrombose) elicited by DMH, R and C toxicities and not as a result of colonic tumor size or metastases. As a single therapy, M appeared to be superior to either R or C alone. However, R + C combination was effective and was further improved upon by its association with M. With the triple combination, (M + R + C), lesions of both cancers decreased in size and/or number and the colon cancer histologically eclipsed from 46% of the treated animals.

Effects of single and combined maltose tetrapalmitate immunotherapy, cyclophosphamide chemotherapy and radiotherapy on ethyl carbamate accelerated primary lung cancer in A/J mice.

A/J mice were given ethyl carbamate to accelerate and to raise to 100 percent the incidence of lung tumours at 34 weeks (day 237) of age. The animals were then divided into groups which received the following treatments: group 1, no treatment; group 2, MTP alone; group 3, radiotherapy alone; group 4, cyclophosphamide alone; group 5, radiotherapy + MTP; group 6, MTP + cyclophosphamide; group 7, radiotherapy followed by cyclophosphamide and group 8, MTP and radiotherapy together followed by MTP and cyclophosphamide. Except for radiotherapy, which was given for 5 consecutive days, MTP and cyclophosphamide were continued till the death of the animals. The treatment efficacies were evaluated by the number and size of tumour nodules, taking into consideration the survival time of the animal. Animals in groups receiving cyclophosphamide died earlier (between days 290 and 315) due to its toxic effects, and half of the radiotherapy-MTP were sacrificed at day 314 for comparison. Although cyclophosphamide alone and radiotherapy plus cyclophosphamide demonstrated antitumour activity, the number of tumour nodules and the nodule diameter were reduced most effectively in group 8 (receiving MTP, radiotherapy and cyclophosphamide). Among the animals in the non-cyclophosphamide group, radiotherapy alone was ineffective. MTP given before and after radiotherapy (group 5) kept tumour volume in control although this group died suddenly. The animals receiving only MTP died between day 430 and 470. The number of tumour nodules and the nodule diameter in the MTP group were, however, significantly reduced when compared to controls or radiotherapy group animals dying at or near the same time.

Mitogenic, immunoadjuvancy, and genetic studies on fatty acyl maltose.

Three synthetic glycolipids, maltose tetrapalmitate (MTP), maltose hexastearate (MHS), and maltose hexalinoleate (MHL) prepared as nontoxic lipid A analogs, and Escherichia coli lipopolysaccharide (LPS) were assayed for their mitogenic activity using spleen lymphocytes in nine inbred mouse strains and three F1 hybrids. The MTP and LPS were also assayed for their ability to enhance plaque-forming cell (PFC) responses using sheep red blood cells as the antigen in the same inbred mouse strains and F1 hybrids, The mitogenic activity of synthetic glyco-lipids was several fold lower than that of LPS and MHL was inferior to MTP and MHS. DBA/2J was the most responsive strain for MTP and DBA/1J and C3H/HeJ the least. The mitogenic activity of MTP was generally in agreement with the PFC response stimulation by it. Low-dose cyclophosphamide treatment of mice synergized MTP for PFC response augmentation. Genetic studies on MTP mitogenicity revealed that 90% of responder DBA/2J X nonresponder C3H/HeJ F1 hybrids had intermediate mitogenic activity. Among F2, 73% had intermediate-high activity and 27% were nonmitogenic. Among F1 X C3H/HeJ backcrosses 11% had high, 56% intermediate, and 33% had no mitogenic activity, whereas, for the F1 X DBA/2J backcross, 14% had high, 36% intermediate, and 50% low or negligible activity. The data favored a single gene for MTP activation of immune cells.

Increased survival of CD1 mice bearing dimethylhydrazine induced primary colon and anal cancers by difluoromethylornithine with concomitant increase in angiosarcoma incidence.

Sixty CDl mice received dimethylhydrazine 20 mg/kg s.c. once weekly for 26 weeks to induce colorectal cancer. At this time the animals harbored frank colorectal cancer and early epidermoid cancer. The animals were divided into six groups that were subjected to the following treatments: none, MTP immunotherapy (MTP) alone, radiotherapy (R) alone, difluoromethylornithine (DFMO) chemotherapy alone and combinations of R+DFMO and R+DFMO+MTP. Criteria of evaluation of treatment efficacy were: number of colorectal tumor lesion and their staging at death, the incidence and size of anal cancer at death and survival time. Radiotherapy alone was marginally effective and MTP treatment was moderately effective in preventing anal cancer and reducing the number of colorectal tumors as well as their size. DFMO was exceptional in preventing anal cancer in a majority of animals and increasing animal survival; the latter effect was due to its preventive action against pyelonephritis, the major cause for animal death. However, in DFMO treated animals, the incidence of angiosarcoma increased from 10-16% (in the absence of DFMO) to 35-50% (in the presence of DFMO). The most effective treatment of the colorectal tumor was the triple combination of R + DFMO + MTP.

Pleomorphic lipoma of the tongue: case report and literature review.

A case of pleomorphic lipoma of the tongue in a 60-year-old woman is reported. Recently described by Enzinger (1977), pleomorphic lipoma is a rare benign tumor, occurring mostly in elderly males. The subcutaneous tissues of the neck and back are the most common sites involved. Differentiation from some histological variants of liposarcoma remains difficult. Complete surgical excision is the treatment of choice. To our knowledge, this is the first case of pleomorphic lipoma of the tongue reported in the English literature.

Combined radiotherapy, chemotherapy, and maltose tetrapalmitate immunotherapy in the treatment of 4'dimethylaminoazobenzene-induced liver cancer.

Therapeutic effects of radiotherapy (R), chemotherapy, and maltose tetrapalmitate (MTP) immunotherapy alone and in combinations were tried against 4' dimethylaminoazobenzene (DAB) induced primary liver cancer in Wistar rats in three separate protocols. Rats were fed a low protein synthetic diet containing 0.06% DAB for 90-120 days. Around 90 days, liver cancers developed in all the animals. In the first protocol, animals were either left untreated or treated with cyclophosphamide (Cy), MTP (i.p. or oral) and Cy plus oral MTP. Rats in the MTP (i.p.) group maintained a steady liver weight but neither Cy nor Cy + MTP influenced the survival time or liver weight. In the second protocol, R as well as a 3-drug combination at 2 dose levels were tried alone and with MTP before or soon after cessation of DAB feeding. Survival times were decreased by R and chemotherapy due to combined toxicities of DAB and treatments and were partially restored by MTP. In the third protocol, MTP, R, and Cy were each tried alone and in combinations, 21 days after cessation of 100-day DAB feeding. Increase in survivals were obtained by each treatment, although tumor weight was best controlled by triple R+ Cy + MTP combination.

Adjuvant immunotherapy of N-[4(5-nitro-2-furyl)-2-thiazolyl]-formamide-induced bladder tumors.

The prophylactic effect of maltose tetrapalmitate (MTP), a newly developed non specific immunoadjuvant in preventing or delaying bladder cancer induction by N-[4(5-nitro-2-furyl)-2-thiazolyl] formamide (FANFT) or in reducing the growth rate of the induced tumor was compared to other well-known immunoadjuvants (BCG, C. parvum, levamisole and pyran copolymer). The evaluation of the prophylactic role of immunoadjuvants demonstrated that MTP, levamisole and C. parvum were the most effective in prolonging animal survival. MTP was found superior to either of them in reducing the tumor size. The development of lung metastasis was lower in the group receiving MTP or C. parvum. Next, MTP was studied for its therapeutic effect against primary FANFT induced tumor. The subcutaneous (s.c.) and oral routes of MTP administration and their combination with intravesical route were tried. Combinations of intravesical with s.c. or oral MTP were most effective in reducing tumor size, obtaining lower metastases along with greater mononuclear lymphocyte infiltration in the primary tumor.

Peutz-Jeghers syndrome. Association of duodenal and bilateral breast cancers in the same patient.

A female patient with Peutz-Jeghers syndrome successively developed bilateral breast carcinoma and malignant transformation of a duodenal hamartomatous polyp, from which she died.