Mutagenicity of pan-fried bovine tissues in relation to their content of creatine, creatinine, monosaccharides and free amino acids

The mutagenicity of pan-fried patties of five bovine tissues (meat, heart, tongue, liver and kidney) containing various concentrations of creatine, monosaccharides and free amino acids were studied. Two experiments were performed, one on single tissues fried at 150, 175 or 200 degrees C for 3 min and the other on mixtures of meat and one of the other four tissues in various proportions, fried at 200 degrees C for 3 min. For both experiments, a double-sided Teflon-coated plate was used. Frying at 150 degrees C induced mutagenicity to Salmonella typhimurium strain TA98 only in the heart sample-6000 revertants/100 gE (grams initial raw weight). Meat, heart and tongue fried at 175 or 200 degrees C showed mutagenicity values between 6000 and 19,600 revertants/100 gE. A linear relationship between mutagenicity and temperature was obtained for each of the three muscles and creatine was converted to creatinine with increasing temperature. Liver or kidney samples fried alone showed insignificant mutagenicity at all three temperatures. The creatine plus creatinine levels of raw meat, heart and tongue samples were between 19 and 33 mumol/g wet tissue. Liver and kidney both showed very low amounts of creatine plus creatinine (about 2 mumol/g wet tissue) in the raw tissue, while free amino acids were high. Glucose levels were high in liver but low in kidney samples. In meat/heart and meat/tongue mixtures the mutagenicity varied between 10,800 and 17,300 revertants/100 gE. The meat/liver and meat/kidney mixtures showed linear relationships between mutagenicity and the proportions of the mixture. The values for the slopes and intercepts of the two lines were almost equal. Among the three groups of precursors (creatine plus creatinine, monosaccharides and free amino acids) the creatine plus creatine in raw tissue seems to be the most important for producing mutagenicity. However, in crusts, the creatinine concentration was the variable with which most of the mutagenicity was associated.     

Inhibitory effect of carbohydrates on the formation of mutagens in fried beef patties.

Beef patties were prepared by mixing minced meat with water and either glucose (1, 2 or 4%), lactose (1, 2 or 4%) or powdered milk (2, 4 or 8%) before frying. In another experiment, minced meat was mixed with starch from golden bread crumbs (3%) or potatoes (4%), with and without glucose (1, 2 or 4%). The patties (100 g) were fried for 3 min at 150 or 180 degrees C in a double-sided fryer. The mutagenic activity in the crust was determined using the Ames test. With the addition of glucose or lactose (1-4%), the mutagenic activity was inhibited by 34-76%. A similar inhibition of the mutagenic activity was obtained with powdered milk. However, starch from golden bread crumbs or potatoes caused only a slight (not significant) decrease in mutagenic activity whereas adding both starch and glucose to the beef patties inhibited mutagenic activity by up to 54%.     

Effects of creatine and creatinine content on the mutagenic activity of meat extracts, bouillons and gravies from different sources

Thirteen commercial meat-flavour samples were analysed for creatine and creatinine content and tested for mutagenicity in the Ames Salmonella/microsome test. In most samples, more than 50% of the creatine had been converted to creatinine. Mutagenicity was related to the creatinine content: 150 mumol creatinine/g dry matter (gdm) gave upwards of 2500 revertants/gdm, concentrations of 15-40 mumol/gdm gave about 100 revertants/gdm and concentrations of 1-10 mumol/gdm gave only low or no significant mutagenicity. No relationship was apparent between coloration and mutagenicity. Beef steaks (initial weight c. 500 g) baked at oven temperatures between 115 and 245 degrees C only showed significant mutagenicity--135 revertants/100 gE (initial raw weight)--in the crust when baked at the highest temperature (245 degrees C). The gravies (meat-juice drip) collected during baking showed a linear increase in mutagenicity with baking temperatures up to 180 degrees C (48-828 revertants/100 gE) and a very sharp increase in mutagenicity for the gravy collected from beef steak baked at 245 degrees C (28,300 revertants/gdm or 19,800 revertants/100 gE). At this high temperature, the brown coloration and the proportion of creatinine to total creatine and creatinine were also dramatically increased, because this gravy dried up completely during the baking procedure.     

Mutational and gene fusion analyses of primary large cell and large cell neuroendocrine lung cancer

Large cell carcinoma with or without neuroendocrine features (LCNEC and LC, respectively) constitutes 3–9% of non-small cell lung cancer but is poorly characterized at the molecular level. Herein we analyzed 41 LC and 32 LCNEC (including 15 previously reported cases) tumors using massive parallel sequencing for mutations in 26 cancer-related genes and gene fusions in ALK, RET, and ROS1. LC patients were additionally subdivided into three immunohistochemistry groups based on positive expression of TTF-1/Napsin A (adenocarcinoma-like, n = 24; 59%), CK5/P40 (squamous-like, n = 5; 12%), or no marker expression (marker-negative, n = 12; 29%). Most common alterations were TP53 (83%), KRAS (22%), MET (12%) mutations in LCs, and TP53 (88%), STK11 (16%), and PTEN (13%) mutations in LCNECs. In general, LCs showed more oncogene mutations compared to LCNECs. Immunomarker stratification of LC revealed oncogene mutations in 63% of adenocarcinoma-like cases, but only in 17% of marker-negative cases. Moreover, marker-negative LCs were associated with inferior overall survival compared with adenocarcinoma-like tumors (p = 0.007). No ALK, RET or ROS1 fusions were detected in LCs or LCNECs. Together, our molecular analyses support that LC and LCNEC tumors follow different tumorigenic paths and that LC may be stratified into molecular subgroups with potential implications for diagnosis, prognostics, and therapy decisions.