Circadian rhythm in toxicities and tissue uptake of 1,2-diamminocyclohexane(trans-1)oxalatoplatinum(II) in mice.

Mechanisms involved in the circadian rhythm in murine tolerance for the new platinum analogue, 1,2-diamminocyclohexane(trans-1)oxalatoplatinum(II) (1-OHP) were sought in 404 male C57BL/6 x DBA/2 F1 mice standardized by 12 h light-12 h dark. A potentially lethal dose of 1-OHP (17 mg/kg i.v.) resulted in 76% long-term survival at 15 h after light onset (HALO) (activity span) as compared to 24% after treatment at 7 HALO (rest span) (chi 2 21.3; P less than 0.001). A total of 204 mice received the same dose of 1-OHP at one of three circadian stages (0, 8, or 16 HALO). No renal toxicity was encountered. Bone marrow and jejunal villi constituted the chief targets of 1-OHP toxicity at this dosage and schedule. Hematological tolerance as gauged by leukocyte counts was optimal when the drug was given at 16 HALO (P from analysis of variance, less than 0.001). Jejunal lesions were less severe after 1-OHP dosing at 16 HALO as compared to 8 HALO (P less than 0.001). Total platinum concentrations were determined in 18 tissues 24 h after 1-OHP dosing. The highest levels of platinum were found in the spleen on day 1 as well as on day 5 following 1-OHP treatment. Despite the fact that the highest platinum concentrations in tissues usually corresponded to drug dosing at 8 HALO, no correlation was documented between such variables and tissue toxicity. Tissue pharmacokinetics of 1-OHP contribute only in part if at all to the circadian rhythm in hematological and jejunal toxicity of this drug.

Hematological toxicity of repeated injections of (chloro-2-ethyl)-ribofuranosyl-3-nitrosourea.

A previous study using a single injection of (chloro-2-ethyl)ribofuranosyl-3-nitrosourea has indicated the low acute hematotoxicity of this nitrosourea. However, because the hematotoxicity of nitrosourea is usually cumulative, we have studied the effect of injecting (chloro-2-ethyl)ribofuranosyl-3-nitrosourea (15 mg/kg) dissolved in 0.2 ml sterile oil C57BL X i.p. into DBA/2F1 mice for 5 consecutive weeks. The dose per injection represents the minimal dose necessary to show the maximal therapeutic efficacy on L1210 leukemia. Bone marrow cellularity and histology, spleen weight, bone marrow and splenic pluripotent stem cells, and colony-forming units committed to granulocyte-macrophage differentiation were measured 1, 2, 4, 7, and 14 days after the last injection in treated and control mice receiving oil only. No morphological or histological changes were found in the spleen and bone marrow of treated mice. In both organs, the number of splenic pluripotent hematopoietic stem cells decreased by about 1 log 1 day after the last injection but rapidly returned to normal values on Days 4 to 14. Granulocyte-macrophage-committed precursors were affected in both organs, at 20% of control values for bone marrow and 5% of control values for spleen on Day 1, with a transient and partial recovery on Day 4 followed by a second drop to 20 and 25% on Day 14. This effect on granulocyte-macrophage precursors contrasts with the absence of significant effect when the same treatment is used on peripheral white blood cell counts. Our results demonstrate that (chloro-2-ethyl)ribofuranosyl-3-nitrosourea belongs to the class of new nitrosoureas with low cumulative hematotoxicity.

A four-day subrenal capsule assay for testing the effectiveness of anticancer drugs against human tumors.

The subrenal capsule assay may predict to which anticancer drug a given patient's tumor is sensitive and may also be used to screen new anticancer drugs. The present study documents that the use of this model requires a histological assessment of both the exploitability of a subrenal capsule assay and the extent of drug-induced antitumor lesions. Thirty-five tumors from 34 patients with solid tumor were submitted to a subrenal capsule assay in a total of 1130 male B6D2F1 mice. After being biopsied, each tumor was dissected by a pathologist and cut into 50 pieces (1.5 X 1.5 X 1.5 cu mm), and one piece was implanted under the renal capsule of 35 mice; the mean tumor diameter was measured on Day 0. Mice were randomized into groups of 6 to 10 animals each. On Days 1, 2, and 3, mice were treated either with placebo (control group) or with various anticancer agents. On Days 4 or 6, mice were sacrificed, the mean tumor diameter measured, and the tumor-bearing kidney fixed in Bouin's picroformol solution and processed for histological analysis after staining with hematein -eosin. Seven histological parameters were blindly rated in a semiquantitative fashion yielding a compound score ( PAPAN ) which estimated the overall quality of each xenograft between -3 and +11. On Day 4, as opposed to Day 6, mean lymphocytic infiltration was 3-fold lower (p less than 0.01), and the rate of xenografts containing well-preserved cancer cells was 2-fold larger (p less than 0.01) in three different tumor specimens. Twenty-two of 31 (71%) assays were evaluable, as defined by a histological quality control test. In those, drug effects were demonstrable by statistically significant differences among groups in 2 assays (9%) by using the relative variation in tumor size as an index of drug effectiveness and in 12 assays (54%) by PAPAN histological score. This suggests the higher sensitivity of histological scoring over tumor size measurements. Moreover, no correlation between relative variation in tumor size and PAPAN was demonstrable with statistical significance indicating the poor reliability of tumor size measurements as an index of the antitumor effectiveness of cytostatic drugs.

Docetaxel chronopharmacology in mice.

Docetaxel tolerance and antitumor efficacy could be enhanced if drug administration was adapted to circadian rhythms. This hypothesis was investigated in seven experiments involving a total of 626 male B6D2F1 mice, synchronized with an alternation of 12 h of light and 12 h of darkness (12:12), after i.v. administration of docetaxel. In experiment (Exp) 1, the drug was given once a week (wk) for 6 wks (20 mg/kg/wk) or for 5 wks (30 mg/kg/wk) at one of six circadian times, during light when mice were resting [3, 7, or 11 hours after light onset (HALO)], or during darkness, when mice were active (15, 19, or 23 HALO). Endpoints were survival and body weight change. In Exp 2 and 3, docetaxel (30 mg/kg/wk) was administered twice, 1 wk apart, at one of four circadian stages (7, 11, 19, or 23 HALO). Endpoints were hematological and intestinal toxicities. In Exp 4, circadian changes in cell cycle phase distribution and BCL-2 immunofluorescence were investigated in bone marrow as possible mechanisms of docetaxel tolerability rhythm. In Exp 5 to 7, docetaxel was administered to mice bearing measurable P03 pancreatic adenocarcinoma (270-370 mg), with tumor weight and survival as endpoints. Mice from Exp 5 and 6 received a weekly schedule of docetaxel at one of six circadian stages (20 or 30 mg/kg/wk at 3, 7, 11, 15, 19, or 23 HALO). In Exp 7, docetaxel (30 mg/kg) was given every 2 days (day 1, 3, 5 schedule) at 7, 11, 19, or 23 HALO. Docetaxel dosing in the second half of darkness (19 or 23 HALO) resulted in significantly worse toxicity than its administration during the light span (3, 7, or 11 HALO). The survival rate ranged from 56.3% in the mice treated at 23 HALO to 93.8 or 87.5% in those injected at 3 or 11 HALO, respectively (Exp 1, P < 0.01). Granulocytopenia at nadir was -49 +/- 14% at 7 HALO compared with -84 +/- 3% at 19 HALO (Exp 2 and 3, P < 0.029), and severe jejunal mucosa necrosis occurred in 5 of 8 mice treated at 23 HALO as opposed to 2 of 18 receiving docetaxel at 7, 11, or 19 HALO (Exp 2 and 3, P < 0.02). The time of least docetaxel toxicity corresponded to the circadian nadir in S or G2-M phase and to the circadian maximum in BCL-2 immunofluorescence in bone marrow. Docetaxel increased the median survival of tumor-bearing mice in a dose-dependent manner (controls: 24 days; 20 mg/kg weekly, 33 days; 30 mg/kg weekly or day 1, 3, 5 schedule, 44 or 46 days, respectively; Exp 5-7). Survival curves of treated mice differed significantly according to dosing time for each dose and schedule (P from log rank <0.003 to P < 0.03). In Exp 5 and 6, the percentage of increase in life span was largest if docetaxel was administered weekly at 7 HALO (20 mg/kg, 220%; 30 mg/kg, 372%) and lowest after docetaxel dosing at 19 HALO (80% with 20 mg/kg) or at 15 HALO (78% with 30 mg/kg). In Exp 7, (day 1, 3, 5 schedule), docetaxel was most active at 11 HALO (percentage increase in life span, 390%) and least active at 23 HALO (210%). Docetaxel tolerability and antitumor efficacy were simultaneously enhanced by drug dosing in the light span, when mice were resting. Mechanisms underlying the tolerability rhythm likely involved the circadian organization of cell cycle regulation. Docetaxel therapeutic index may be improved with an administration at night in cancer patients, when fewest bone marrow cells are in S or G2-M phase.

HIV-associated endometritis.

Using immunohistochemical staining, in situ hybridization and a combination of both, we demonstrate here the replication of HIV in the endometrial stroma. Infected cells do not belong to the T-lymphocyte lineage but rather to a monocyte-macrophage cell type. This report suggests a possible relationship between HIV infection and endometritis. Moreover, HIV replication in endometrial tissues could play a role in heterosexual and materno-fetal transmission.

Experimental protection by a luteinizing hormone-releasing hormone agonist of a bone marrow nitrosourea aplasia. Preliminary observations.

D-Tryptophan-6-luteinizing hormone-releasing hormone (D-Trp6-LH-RH) applied from day -8 to day +15 before and after administration of the nitrosourea analog, N,N'-bis[N-(2-chloroethyl)-N-nitrosocarbamoyl]cysteamine (CNCC), favored bone marrow restoration, as independently evaluated by 3 observers in a double-blind fashion.

Light chain deposition disease with liver dysfunction.

Light and transmission electron microscopic study as well as immunohistochemical investigation were performed on three cases of light chain deposition disease (LCDD) with severe liver dysfunction. In two cases, the amount of light chain deposits in the liver was moderate and did not correlate with the severity of clinical and biological symptoms. Ultrastructural study demonstrated a collagenization of the Disse's space, with basement membrane-like material in association with light chain deposits. Immunohistochemical investigation showed a marked increase of collagen types I, III, and IV, as well as fibronectin and laminin in perisinusoidal space. This study suggests that collagenization of the Disse's space has a minor role in liver dysfunction. The analogy between kidney and liver lesions in diabetes and in LCDD is stressed, but the mechanism of this abnormal accumulation of matrix proteins remains unknown.

Expression of histocompatibility antigens and characterization of the lymphocyte infiltrate in hyperplastic polyps of the large bowel.

HLA-DR expression, lymphocyte subsets, and the distribution of proliferating cells were studied in hyperplastic polyps from the colorectum. The density of T-cells (CD5+) (mean of cells/mm2 of tissue +/- SEM) was higher in the lamina propria of hyperplastic polyps (64.2 +/- 4.2) than in normal colonic mucosa (36.7 +/- 2.6, P less than .001). The CD4/CD8 ratio was higher in hyperplastic polyps (6.3 +/- 0.9, P less than .0001) and in colonic adenomas (5.9 +/- 0.9, P less than .001) compared with normal mucosa (2.3 +/- 0.2). Lymphocytes of the lamina propria were never Ki-67 positive either in normal mucosa or in hyperplastic polyps or adenomas. The epithelial layer of hyperplastic polyps and of normal mucosa did not express the HLA-DR antigen, whereas pericryptal fibroblasts and most of the leukocytes of the lamina propria were strongly positive for this antigen. In the epithelial layer proliferating cells were localized exclusively in the lower part of epithelial crypts, as was the case in normal mucosa, whereas in adenomas Ki-67-positive cells were present throughout the entire height of the mucosa. Thus, in hyperplastic polyps lymphocytes are increased in the lamina propria, with a predominance of the CD4 subset in close contact with HLA-DR positive pericryptal fibroblasts.

Lymphocyte subsets and HLA-DR expression in normal pancreas and chronic pancreatitis.

In an attempt to study the mechanisms leading to fibrosis in chronic pancreatitis, an in situ immunohistochemical investigation of lymphocytes and of class II major histocompatibility complex expression (HLA-DR) by epithelial cells has been designed. Samples of normal pancreas (n = 8), chronic calcifying pancreatitis (n = 4), chronic obstructive pancreatitis (n = 6), and diffuse fibrosing pancreatitis (n = 6) have been studied. In normal pancreas, T-lymphocytes were rare and were located in the epithelial layer of pancreatic ducts and in the periductal connective tissue. Duct cells were constantly HLA-DR negative. In chronic calcifying pancreatitis and chronic obstructive pancreatitis, T cells were numerous and were located around ducts and in the spreading areas of fibrous septa. In chronic obstructive pancreatitis, the duct cells strongly expressed the HLA-DR antigen. In diffuse fibrosing pancreatitis, fibrous tissue was devoid of lymphocytes and duct cells never expressed the HLA class II antigen. These results suggest that lymphocytes are involved in the fibrosing process occurring in chronic calcifying pancreatitis and chronic obstructive pancreatitis but not in diffuse fibrosing pancreatitis. The significance of de novo expression of HLA-DR antigen by duct cells is discussed.

Serum apolipoprotein A-I in alcoholic patients with chronic calcifying pancreatitis.

Serum apolipoprotein A-I measurement was compared in alcoholic patients according to presence or absence of chronic pancreatitis and liver fibrosis. Among alcoholic patients without liver disease, apolipoprotein A-I was significantly lower in patients with chronic pancreatitis (157 +/- 70 mg/dl) than in patients without pancreatitis (209 +/- 74 mg/dl, p less than 0.001). In cirrhotic patients, apolipoprotein A-I was lower in patients with chronic pancreatitis (82 +/- 35 mg/dl) than in patients without pancreatitis (102 +/- 45 mg/dl), but this difference was not significant. The decrease of serum apolipoprotein A-I was independent of nutritional parameters whether or not there was cirrhosis. Immunohistochemical study of pancreatic samples with chronic pancreatitis showed that apolipoprotein A-I was located in the pancreatic fibrosis whereas lobules were unstained. This study suggests that apolipoprotein A-I is trapped by the pancreatic extracellular matrix and that this sequestration might explain, in part, the decrease of the serum apolipoprotein A-I.

Localization of apolipoprotein A-I and apolipoprotein A-II in human atherosclerotic arteries.

Apolipoprotein A-I and apolipoprotein A-II, the two major protein components of the high-density lipoproteins, were visualized in human arteries using an immunofluorescence technique. Apolipoprotein A-I and apolipoprotein A-II were codeposited into the intima and upper media of normal arteries of atherosclerotic patients. The amount of deposits increased in fatty streaks. In atherosclerotic plaques, apolipoproteins accumulated around the necrotic material. These two apoproteins were present in the extracellular matrix as well as in the foam cells surrounding the atherosclerotic lesions. The concomitant intracellular localization of apolipoprotein A-I and of apolipoprotein A-II in the cytoplasm of foam cells supports the hypothesis that extracellular high-density lipoprotein particles are internalized in the macrophages during the atheromatous process.

Carcinosarcoma of the liver with mesenchymal differentiation: a case report.

Carcinosarcoma of the liver with mesenchymal differentiation are very rare in adult patients. A case is reported with an exhaustive pathologic examination and review of the literature. A 61-year old man presented with general fatigue and dull abdominal pain. Two liver masses were diagnosed and resected by a right hepatectomy. Specimen pathology revealed that the tumor and lymph node consisted of two cancerous components. One carcinomatous component corresponding to a hepatocellular carcinoma and a sarcomatous component characterized by a diffuse proliferation of spindle shaped cells with chondrosarcomatous and osteosarcomatous changes. Patient died 9 months later of a diffusion of the tumor. For the first time, to our knowledge, a mesenchymal differentiation is demonstrated in liver carcinosarcoma.

Modulation of cisplatin chronotoxicity related to reduced glutathione in mice.

Intracellular reduced glutathione (GSH) concentrations were measured according to the tissue sampling-time along the 24 h scale in male B6D2F1 mice. A significant circadian rhythm in GSH content was statistically validated in liver, jejunum, colon and bone-marrow (P < or = 0.02) but not in kidney. Tissue GSH concentration increased in the dark-activity span and decreased in the light-rest span of mice. The minimum and maximum of tissue GSH content corresponded respectively to the maximum and minimum of cisplatin (CDDP) toxicity. The role of GSH rhythms with regard to CDDP toxicity was investigated, using a specific inhibitor of GSH biosynthesis, buthionine sulfoximine (BSO). Its effects were assessed on both tissue GSH levels and CDDP toxicity at three circadian times. BSO resulted in a 10-fold decrease of the 24 h-mean GSH in kidney. However a moderate GSH decrease characterized liver (-23%) and jejunum (-30%). BSO pretreatment largely enhanced CDDP toxicity which varied according to a circadian rhythm. Although BSO partly and/or totally abolished the tissue GSH rhythms, it did not modify those in CDDP toxicity. We conclude that GSH have an important influence on CDDP toxicity but not in the circadian mechanism of such platinum chronotoxicity.

[Adenocarcinoma in heterotopic gastric pancreas]. / Adénocarcinome développé sur un pancréas ectopique gastrique.

The development of an adenocarcinoma in heterotopic pancreas of the stomach is uncommon and its diagnosis is difficult. We report the case of a 42 year-old man who was admitted for gastric stasis, related to an antral tumour. He had been treated for Hodgkin's disease by radiation therapy. 16 years ago. Gastrectomy was performed. On light microscopy, the gastric wall was infiltrated by pancreatic ectopic tissue, normal in the submucosa, but dedifferentiated with cytological and architectural criteria of malignancy in the muscular and the serous layers. Malignant transformation of an ectopic pancreas is theoretically possible but is very uncommon. In our case, the role of previous radiation therapy as a promoting factor is discussed.

[Hyperplastic colorectal polyp: histological, histochemical and immunohistochemical study of 43 cases]. / Le polype hyperplasique colo-rectal: étude histologique, histochimique et immunohistochimique de 43 cas.

Forty-three solitary hyperplastic polyps removed from the colon or the rectum (HPC) were examined under light microscopy. A histochemical and immunohistochemical study was undertaken in order to evaluate semiquantitatively the nature and the distribution of epithelial mucins and the secretion of carcinoembryonic antigen (CEA). Ten HPC had a peculiar morphologic pattern (four with regenerative dysplasia and six with adenomatous foci). CEA secretion was always increased (37 per cent of cases) or highly increased (63 per cent of cases) with respect to the normal colonic mucosa. The nature and distribution of the secreted acid mucins were modified: sulfomucin was equal (25 per cent of cases) or higher (75 per cent of cases) than that in normal rectal or sigmoid colonic mucosae; sialomucin was strongly decreased in 91 per cent of cases. Some of these functional changes (CEA) are also observed in neoplastic lesions. These findings are not in accord with the hypothesis that hyperplastic polyp is a simple hyperplasia of the mucosal epithelium and suggest a disorder in cellular differentiation, particularly for the larger polyps.

[Ciliated cysts of the liver. 2 cases]. / Kystes à revêtement cilié du foie. Deux cas.

We report herein two cases of ciliated hepatic cysts. These exceptional lesions belong to the category of solitary nonparasitic cysts. They are probably dysembryoplastic and appear to derive from the embryonic foregut. Diagnosis is based on microscopic examination of the surgical specimen. The role of aspiration cytology in the preoperative diagnosis of this lesion has not yet been defined.

[Predictive risk factors for progression to cirrhosis in early stage alcoholic liver disease]. / Facteurs prédictifs d'évolution vers la cirrhose à un stade précoce de la maladie alcoolique du foie.

OBJECTIVE: To assess the risk factors of cirrhosis in early stage alcoholic liver disease. PATIENTS: We investigated 83 heavy drinkers (60 males and 23 females) in whom the first of two liver biopsies showed normal or pure alcoholic fatty liver. RESULTS: When the six following variables: sex, age, delay between the first and last biopsy, total duration of alcohol consumption before the first biopsy, daily alcohol consumption for the last 5 years before the first biopsy and the extent of fatty liver in the first biopsy, were considered together in stepwise regression analysis, the delay between the first and last biopsy (p < 0.0001), sex (P < 0.004) and the extent of fatty liver in the first biopsy (P < 0.06) significantly improved the prediction of cirrhosis. The odds ratio of cirrhosis for a women was 19.1 (confidence interval 95% [1.85-197]). The odds ratio for cirrhosis for a percentage of fatty liver > or = 5/10 was 7.4 (confidence interval 95% [1-92]). CONCLUSION: With the same delay between two liver biopsies, the female sex and the extent of fatty liver are two independent risk factors for the development of cirrhosis in heavy drinkers.

Efficacy of long term recombinant interferon-alfa in patients with chronic hepatitis C. A clinical, biological, histological and immunohistological study.

A prospective, non randomized trial was conducted in 22 patients with chronic hepatitis C treated for 6 months with interferon-alfa at the dose of 3 million units three times a week followed by decreasing doses for the following 6 months. Transaminase activity measurements were taken at 2, 6, 12, 18, and 24 months and a liver biopsy was performed at zero and 6 months to assess efficacy. Immunohistochemical study of HLA class I and lymphocyte subsets was performed on the biopsy specimen before and after 6 months of treatment. A complete biological response was observed in 82, 82, 60, 64 and 70 percent of patients at 2, 6, 12, 24 and 36 months, respectively. Four patients relapsed including 2 patients who suddenly stopped taking interferon-alfa. An improvement of histological features was observed in 90 percent of patients for necrosis and in 80 percent of patients for inflammation, with an overall improvement in Knodell's score reaching 85 percent. Immunohistochemical studies showed CD8+ cell infiltration already present in the area of necrosis before treatment. These tests also showed that HLA class I expression by the hepatocyte membranes was increased at 6 months and was identical in patients with or without histological improvement.

[Aortic dissection and Horton arteritis]. / Dissection aortique et artérite de Horton.

Aortic dissection in a 61-year-old woman treated symptomatically and with a fatal outcome within three months by extension to the arch of the aorta and its major vessels, and as a direct result of haemopericardium. The association with temporal arteritis, presenting before the aortic complication by prolonged headache with raised sedimentation rate and painful inflammation of both temporal arteries, was confirmed by histopathological examination of the aorta. This revealed giant cell arteritis independent of atheromatous lesions. The association of aortic dissection and temporal arteritis is very rare. Clinical and pathological examination of the smaller arteries and in particular the craniocephalic arteries is therefore justified in all cases of aortic dissection. When it affects the aorta, temporal arteritis may favor aortic dissection as a result of changes in the media.

[Polyps and glandular polyposis of the fundus. Study of 20 cases and review of the literature]. / Polype et polypose glandulo-kystique du fundus. Etude de 20 cas et revue de la littérature.

An histological and histochemical study has been performed in 20 cases of fundic gland polyps. In five cases, polyps were numerous and associated, in two cases, with a familial adenomatosis coli. The histological, histochemical characteristics and the study of semi-serial sections suggest that fundic gland polyp is a focal distension of the fundic tube without evidence of an hamartomatous neither a neoplastic origin. Because of a possible association with a familial colonic polyposis, an endoscopical investigation is usefull, specially in fundic gland polyposis.