The cytokine microenvironment of human colon carcinoma. Lymphocyte expression of tumor necrosis factor-alpha and interleukin-4 predicts improved survival.

BACKGROUND: The functional significance of cytokines expressed in situ by tumor cells and tumor infiltrating lymphocytes (TIL) in human colon carcinomas is largely unknown. METHODS: We assessed TIL expression of interleukin-2 (IL-2), IL-4, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), granulocyte-macrophage colony stimulating factor (GM-CSF), IL-10, and transforming growth factor-beta (TGF-beta), and tumor cell expression of IL-10 and TGF-beta in situ in 49 primary colon carcinomas and 20 metastases using immunohistochemistry. RESULTS: The percentage of primary colon carcinoma samples in which > 20% of TIL expressed each cytokine was as follows: IL-4: 47%; TNF-alpha: 22%; TGF-beta: 10%; IFN-gamma: 6%; IL-2:2%; IL-10: 0%; and GM-CSF: 0%. Lymphocytes more commonly infiltrated colon carcinoma primaries than metastases, and TIL expression of IL-4 and TNF-alpha was more common in primary than metastastatic carcinomas. Expression of TNF-alpha by even a small proportion (> or = 3%) of the TIL in a colon carcinoma specimen was associated with better overall survival (P = 0.01) when compared with patients with little or no TIL TNF-alpha expression (5-year survival 82% vs. 47%). Expression of IL-4 by > or = 20% of colon carcinoma TIL was also associated with improved survival (P = 0.01; 5-year survival 87% vs. 50%). The expression of IL-10 or TGF-beta by colon carcinoma TIL or colon tumor cells themselves was not associated with impaired survival. Benign epithelial cells stained positively for IL-10 and TGF-beta more frequently than tumor cells (P < 0.001). CONCLUSIONS: There are differences between the immune microenvironment of primary tumors and metastases. Although IL-10 is expressed by colon carcinoma cells and TIL, it is unlikely that it plays an important immunosuppressive role. TNF-alpha and IL-4 are commonly expressed by colon carcinoma TIL and both are associated with improved survival.

In situ cytokine production by breast cancer tumor-infiltrating lymphocytes.

BACKGROUND: Human breast cancers progressively grow despite the presence of extensive lymphocytic infiltration and specific antitumor immune recognition, thereby calling into question the competency of breast tumor-infiltrating lymphocytes (TIL). The function of breast TILs in vivo and their possible role in the suppression of an antitumor immune response are largely unknown. METHODS: The cytokines produced in situ by lymphocytes in 89 breast carcinomas and 14 benign breast lesions were assessed using immunohistochemistry. RESULTS: The majority of tumor and benign breast samples contained T-cell infiltrates, which were disclosed using an anti-CD3 antibody stain. The percentage of tumor samples in which > or =3% of the lymphocytes were producing cytokines was as follows: interleukin (IL)-2 45%, IL-4 36%, tumor necrosis factor-alpha (TNF-alpha) 28%, transforming growth factor-beta 1 (TGF-beta 1) 20%, IL-10 11%, interferon-gamma (IFN-gamma) 4%, and granulocyte-macrophage colony-stimulating factor (GM-CSF) 3%. Production of IL-2, IL-4, and TGF-beta 1 by TILs in breast cancers exceeded that detected in benign breast lesions (p < 0.005). Significantly more tumor samples contained lymphocytes producing IL-2, IL-4, TGF-beta 1, and TNF-alpha than IFN-gamma and GM-CSF (p < 0.002 for each comparison). One or more of the potentially immunoinhibitory cytokines-IL-4, IL-10, or TGF-beta 1-were produced by lymphocytes in 44% of the specimens. No significant associations were seen between lymphocyte production of a particular cytokine and disease-free survival (median follow-up 43 months). CONCLUSIONS: Immunohistochemical techniques can be used to detect cytokine secretion by TILs in preserved tissue. The relative lack of secretion of IFN-gamma and GM-CSF, rather than a deficiency of IL-2, may explain why the antitumor immune response to breast cancer is impaired.

Investigation of a biodegradable, implantable antibiotic delivery system on rate of wound infection.

Biodel, a bioabsorbable polymer, may be adapted to serve as an antibiotic delivery system for high local concentrations of drugs. In an autocontaminated, irradiated wound model with a high intrinsic infection rate, implanted gentamicin-impregnated Biodel beads prevented wound infections in the submental area of rats.

The isolation and identification of steroidal sapogenins in Kleingrass.

Kleingrass (Panicum coloratum L) has been reported to cause hepatogenous photosensitization in sheep and goats in West Texas since 1973. The toxin in Kleingrass has been suspected of being a steroidal saponin as Kleingrass produced characteristic birefringent crystals similar to those produced by saponin-containing plants such as Agave lecheguilla and Tribulus terrestris. In this present study, steroidal saponis were isolated from Kleingrass and their sapogenins were identified as diosgenin and yamogenin by means of thin-layer chromatography and infrared spectrophotometric analysis.

The developmental toxicity of an isolate from the plant Solanum dimidiatum (potato-weed) in Syrian golden hamsters.

The developmental toxicity of an ethanol extract from Solanum dimidiatum was investigated in date-bred Syrian golden hamsters. Neural tube defects were seen in three Syrian golden hamster fetuses, from one litter out of 12, at a maternal dose of 1.52 mg of plant isolate per g of body weight. Other malformations observed were syndactyly in a single fetus, from one litter out of ten, in the 0.45 mg/g dose group and gastroschisis in a single fetus, from one litter out of nine, in the 1.01 mg/g dose group. These findings suggest minimal developmental toxicity for this plant isolate.

Collection of pheromone from atmosphere surrounding boll weevils,Anthonomus grandis.

An effluvial method was developed to collect the pheromone, grandlure from actively calling male boll weevils,Anthonomus grandis Boheman. The adsorbant, Porapak Q (ethylvinylbenzene-divinylbenzene), was utilized to trap and concentrate the pheromone. Captured pheromone was desorbed from columns packed with Porapak Q by elution withn-pentane and quantified by capillary column gas-liquid chromatography. In recovery studies with known amounts of synthetic grandlure, we found that the amount of each pheromone component collected was a function of collection duration, elution volume, and initial concentration. This effluvial method was capable of recovering as much as 94.9% of a known quantity (80 µg) of grandlure. The chromatograms were free of extraneous peaks. In studies of insect-produced pheromone, the effluvial method was used to collect pheromone from the air space surrounding male boll weevils as they fed on flower buds from CAMD-E cotton. The quantity and quality of boll-weevil-produced pheromone was determined for days 6, 8, 10, 11, 12, 13, and 14 of boll weevil adulthood. The maximum quantity of natural pheromone was produced on day 13 (4.2 µg/weevil) with a pheromone component ratio of 2.41∶2.29∶0.95∶1 for components I, II, III, and IV, respectively. The effluvial method described in this report is an efficient method to collect and quantify boll weevil pheromone from the atmosphere surrounding actively calling insects. Other applications of this method are suggested.

Kleingrass (Panicum coloratum L.) poisoning in sheep.

Twenty-four lambs grazing pastures of Panicum coloratum developed photosensitization secondary to hepatic dysfunction. Lesions were necrosis of scattered hepatocytes, obstruction of small bile ducts and bile canaliculi by small aggregates of birefringent crystals, and accumulation of birefringent crystals in phagocytes within sinusoids. The number of crystals in livers of affected sheep varied, depending on the amount of time of exposure to toxic plants and severity of hepatic abnormalities. Crystals in the liver were soluble in acidified ethyl alcohol, acetic acid, pyridine, chloral hydrate, and methanol, but not in xylene, petroleum ether, diethyl ether, acetone, water, or cold ethyl alcohol. Crystals were not stained by oil red O. There was necrosis of epithelial cells in renal distal convoluted tubules, papillary muscles of the heart, and the adrenal cortex. Lesions of Panicum coloratum-associated disease are similar to those associated with photosensitization induced by Tribulus terrestris, Agave lecheguilla, and Nolina texana.

In vivo effects of hymenoxon on glucose metabolism in rat brain.

Hymenoxon inhibition of brain hexokinase resulted in inhibition of glucose utilization evidenced by a decrease in pyruvate and the accumulation of free glucose in the brains of the in vivo treated rats. Inhibition of cerebral energy processes, resulting from inhibition of glycolysis, eventuated the coma and death of one rat at 13 hr. It also accounted for the depression, observed to increase with hymenoxon exposure time, in the treated rats. The increased sulfhydryl group concentration in the brains of the treated rats suggested that increased concentration of reduced glutathione may have resulted from a decrease in its oxidation caused by hymenoxon interaction with sulfhydryl of endogenous brain constituents.

Sporidesmin toxicity in rabbits: biochemical and morphological changes.

The earliest lesion in rabbits dosed orally with 2 mg of sporidesmin per kg of body weight was necrosis of occasional hepatocytes 1 day after dosing. The most consistent lesion was a severe necrotizing cholangitis of medium and large-sized intrahepatic and extrahepatic bile ducts, first seen 2 days after dosing. Similar lesions were also present in the gall bladder of some rabbits. Expansion of portal triads with fibrous tissue and proliferating bile ductules progressed to pseudo-lobulation by 21 days. Other hepatic changes observed irregularly included large infarcts at the periphery of some lobes, and multiple small foci of coagulation necrosis in midzonal and periportal regions. Vascular necrosis and thrombosis, invariably adjacent to necrotic bile ducts, was presumably responsible for the hepatic necrosis. Serum cholesterol and total bilirubin concentrations and GGT activity reached peaks 15 days after dosing and were useful indicators of the severity of biliary lesions. Serum ID activity was the most useful indicator of hepatic necrosis following oral dosing with sporidesmin. The similarity between hepatobiliary lesions observed in sheep and rabbits with experimental sporidesmin toxicity suggests that the rabbit would be a useful model for studying methods of treatment and prevention of "facial eczema" in ruminants.

Absence of sporidesmin production by twelve Texas isolates of Pithomyces spp.

Twelve isolates of Pithomyces spp. from Texas were tested for sporidesmin toxin production, using both high-performance and thin-layer chromatography techniques. None of the Texas isolates produced the toxin under the conditions used. A control toxigenic New Zealand isolate, Pithomyces chartarum strain C, was grown simultaneously under the conditions tested and was found to produce sporidesmin in all cases.

The antagonistic effect of L-cysteine in experimental hymenoxon intoxication in sheep.

L-Cysteine was given IV at dosages of 34.5, 48.8, 69.0, 97.6, or 138.0 mg/kg of body weight to groups of sheep previously treated (10.6 minutes) intraperitoneally with an LD92 of hymenoxon (12.8 mg/kg). L-Cysteine at dosages of 69.0, 97.6, and 138.0 mg/kg significantly increased survival rate 8.4-, 10-, and 12.5-fold, respectively, over that of gymenoxon-treated sheep not given L-cysteine. L-Cysteine at dosages of 34.5 and 48.8 mg/kg failed to significantly reduce mortality. Surviving hymenoxon-treated sheep given 97.6 or 138.0 mg of L-cysteine/kg showed mild or no clinical signs of toxicosis. There was no dosage of L-cysteine that significantly changed the survival time for fatally intoxicated sheep.

The effects of hemolyzed blood on pulmonary and systemic arterial pressure and heart rate of the dog.

The injection of hemolyzed blood into the venous system of the dog produced a rise in the pulmonary arterial pressure and a fall in the systemic arterial pressure. There were variable changes in the heart rate. Comparison with serotonin injections indicated that the above responses were produced by factors other than serotonin.