Many cancer cells display the Warburg effect, that is, enhanced glycolysis followed by fermentation (conversion of pyruvate to lactate). Recently, the molecular basis for these effects has started to be elucidated, and the up-regulation of the lactate dehydrogenase A (LDH-A) isoform of lactate dehydrogenase is felt to be a major molecular mediator of this phenomenon. Moreover, LDH-A expression in tumor tissue and LDH-A levels in blood portend a bad prognosis, and LDH-A blockade can lead to tumor growth inhibition in tumor transplant models. We have extended existing data (some of which were published during the time when we were carrying out our studies) in two important ways: 1) inhibition of LDH-A in a glycolytic lung cancer cell line results in reactive oxygen species-mediated apoptosis and increased sensitivity to the chemotherapeutic drug paclitaxel and 2) inhibition of fermentative glycolysis can also be accomplished by activation of the pyruvate dehydrogenase complex by the drug dichloroacetate, now undergoing clinical trials, and that this phenomenon can be monitored in vivo in a noninvasive real-time manner through magnetic resonance spectroscopy using hyperpolarized pyruvate. Collectively, these data suggest that in vivo effects of drugs that redirect the fate of pyruvate, and hence are aimed at reversing the Warburg effect, could be monitored through the use of hyperpolarized magnetic resonance spectroscopy, a method that is scalable to human use.

LEC10 Chinese hamster ovary (CHO) cells are gain-of-function mutants that express N-acetylglucosaminyltransferase III (GlcNAc-TIII), the glycosyltransferase that adds the bisecting GlcNAc to complex N-glycans. LEC10 cells are useful for glycosylation engineering of recombinant glycoproteins, including antibody therapeutics, for defining lectin recognition specificities and for determining biological functions of the bisecting GlcNAc. We show that three CHO mutants, LEC10, LEC10A, and LEC10B, arose due to transcriptional activation of the quiescent CHO Mgat3 gene. They each express Mgat3 gene transcripts of approximately 4.7 kb at different levels (LEC10B > LEC10 > LEC10A). Southern analyses gave a single band in LEC10, LEC10A, and parent CHO DNA with four restriction enzymes but an additional band with three of them in LEC10B genomic DNA, indicative of a duplication event in LEC10B. The deduced amino acid sequence of the Mgat3 gene expressed in each CHO mutant and in parent CHO genomic DNA is identical. However, 5' UTR sequences differ with LEC10 and LEC10B containing a 5' UTR segment of the Atf4 gene downstream of the Mgat3 gene in human and mouse. Somatic cell hybrid analysis indicated that the LEC10B Mgat3 gene was induced by a cis mechanism. LEC10B glycoproteins bound more erythroagglutinin lectin (E-PHA) than LEC10 glycoproteins and MALDI-TOF MS revealed a broad spectrum of complex, bisected N-glycans expressed by the LEC10B mutant. LEC10B is therefore the cell line of choice for producing recombinant glycoproteins carrying bisected N-glycans and for investigating biological functions of the bisecting GlcNAc.

Melanoma is usually driven by mutations in BRAF or NRAS, which trigger hyperactivation of MAPK signaling. However, MAPK-targeted therapies are not sustainably effective in most patients. Accordingly, characterizing mechanisms that co-operatively drive melanoma progression is key to improving patient outcomes. One possible mechanism is the Hippo signaling pathway, which regulates cancer progression via its central oncoproteins YAP and TAZ, although is thought to be only rarely affected by direct mutation. As YAP hyperactivation occurs in uveal melanoma, we investigated this oncogene in cutaneous melanoma. YAP protein expression was elevated in most benign nevi and primary cutaneous melanomas but present at only very low levels in normal melanocytes. In patient-derived xenografts and melanoma cell lines, we observed variable reliance of cell viability on Hippo pathway signaling that was independent of TAZ activity and also of classical melanoma driver mutations such as BRAF and NRAS. Finally, in genotyping studies of melanoma, we observed the first ever hyperactivating YAP mutations in a human cancer, manifest as seven distinct missense point mutations that caused serine to alanine transpositions. Strikingly, these mutate four serine residues known to be targeted by the Hippo pathway and we show that they lead to hyperactivation of YAP.Implications: Our studies highlight the YAP oncoprotein as a potential therapeutic target in select subgroups of melanoma patients, although successful treatment with anti-YAP therapies will depend on identification of biomarkers additional to YAP protein expression. Mutations and methylation changes in Hippo pathway genes in the TCGA melanoma cohort. A, Somatic coding mutations (top) and CN changes (bottom) affecting Hippo pathway genes identified in patients from the TCGA cohort (56), organized horizontally according to the BRAF-, NRAS-, NF1-, TP53-, RAC1-or PTEN-mutant melanoma subtype of each patient and vertically according the mutation frequency in each Hippo pathway gene. Mutations colored by type: amp, amplification; hemi, hemizygous; homo, homozygous; del, deletion. B, top: distributions of methylation values across 473 TCGA patients in 18 known/predicted oncogenes and 33 known/predicted tumor suppressor genes in the Hippo pathway. Bottom: Spearman coefficient values for correlations between methylation and mRNA expression for each gene. C, Methylation versus mRNA expression plots for YAP and TAZ. Each dot represents values for an individual patient. Blue dots, mutated gene; red dots, wild-type gene. r is the Spearman correlation coefficient; Ã , P < 0.001. YAP and Melanoma

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Testing of sequential sludge washing in triplicate using typical biosurfactant saponin was conducted to remove heavy metals.

The rich and delicate flavors are the mam reason that Chinese cuisine is held in high esteem worldwide. Seasonings and spices are heavily used in most Chinese foods. This review summarizes the flavor chemistry of some seasonings and spices such as soy sauce, vinegar garlic, scallions, 254 HO ET AL.shallots, ginger, star anise, and black mushroom, all of which are commonly used in Chinese foods. The possible interactions of seasonings or spices with other ingredients in the cooking of Chinese dishes are emphasized. Selected examples of the flavors of traditional Chinese foods are presented.The Chinese have developed one of the most sophisticated cuisines in the world. Every day a billion mouths help to make Chinese food better and better. While it is often said that a common written language holds the Chinese civilization together, chopsticks, perhaps, play a stronger role as "cultural cement" than the Chinese writing brush (1).The rich and delicate flavors are the main reason that Chinese" cuisine is so well liked and popular. Seasonings and spices are heavily used in most Chinese foods. Explicit recommendations for the use of seasoning appear in Li Ji (The Book of Rites), believed to have been written in the Spring-Autumn Era (before 476 B.C.). For instances:For seasoning, mostly sourness in the spring, mostly bitterness in the summer, mostly spiciness in the autumn, mostly saltiness in the winter, moderated always with smooth and sweet seasonings.... (1) Climate has a considerable influence on the seasonings favored in any particular geographical area. The people of Sichuan and Hunan, situated in the Yangtze River Basin, use chili peppers with a heavy hand in their cooking; in the Yellow River Basin areas, large quantities of scallion and garlic are consumed; and in the coastal provinces of Guandong, Fujian, and Taiwan, ginger root is preferred.An attempt is made here to review the flavor chemistry of some seasonings and spices commonly used in Chinese foods. Special efforts are paid to discussion of the possible interactions of seasonings or spices with other ingredients in the cooking of Chinese dishes. Selected examples of the flavors of traditional Chinese foods are also presented. SOY SAUCE FLAVORSoy sauce is the most popular condiment used in Chinese cuisine. Many Chinese consider their food "naked" without the presence of soy sauce. In the production of genuine fermented soy sauce, heat-treated soybean and wheat are cultured with Aspergillus oryxae or sojae and then mixed with salt water to make a mash. The mash is fermented with lactobacilli and yeast, and then is well aged to make soy sauce (2).

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Hyperoside (quercetin-3-O-D-galactoside) is a flavonol glycoside which has been isolated from different plants. It has different pharmacological actions such as anti -inflammatory, anti-depressant, neuroprotective, cardio-protective, anti-diabetic, anti-cancer, anti-fungal, radio-protective, gastro-protective, and antioxidant activities. Studies on its pharmacokinetic (PK) properties revealed that it is a stable compound with no significant gender variation in its activities. Other significant details on its pharmacological properties and information for future investigations on its components are provided.