Duplication polymorphisms in exon 4 of κ-casein gene in yak breeds/populations.

The objective of this study was to compare 12 bp-duplication polymorphisms in exon 4 of the κ-casein gene among 3 breeds/populations of yak (Bos grunniens). Genomic DNA was extracted from yak blood or muscle samples (N = 211) and a partial sequence of exon 4 of κ-casein gene was amplified by polymerase chain reaction. A polyacrylamide gel electrophoresis assay of the products (169 bp) revealed 2 variants. These variants differed in a 12-bp duplication of the nucleotide sequence corresponding to amino acids 147-150 (Glu-Ala-Ser-Pro) or 148-151 (Ala-Ser-Pro-Glu). The genotype frequency and gene frequency of the 2 κ-casein variants differed among the 3 yak breeds/populations. The long form of the κ-casein gene was the predominant allele, and the Jiulong yak showed the highest frequency of the short form variant of the κ-casein gene. In addition, 2 nucleotide differences resulting in amino acid substitutions were also identified in yaks. These results are significant for designing a breeding strategy to improve the genetic makeup of yak herds.

Small nucleolar RNA 42 acts as an oncogene in lung tumorigenesis.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer death, reflecting the need for better understanding the oncogenesis, and developing new diagnostic and therapeutic targets for the malignancy. Emerging evidence suggests that small nucleolar RNAs (snoRNAs) have malfunctioning roles in tumorigenesis. Our recent study demonstrated that small nucleolar RNA 42 (SNORA42) was overexpressed in lung tumors. Here, we investigate the role of SNORA42 in tumorigenesis of NSCLC. We simultaneously assess genomic dosages and expression levels of SNORA42 and its host gene, KIAA0907, in 10 NSCLC cell lines and a human bronchial epithelial cell line. We then determine in vitro functional significance of SNORA42 in lung cancer cell lines through gain- and loss-of-function analyses. We also inoculate cancer cells with SNORA42-siRNA into mice through either tail vein or subcutaneous injection. We finally evaluate expression level of SNORA42 on frozen surgically resected lung tumor tissues of 64 patients with stage I NSCLC by using quantitative reverse transcriptase PCR assay. Genomic amplification and associated high expression of SNORA42 rather than KIAA0907 are frequently observed in lung cancer cells, suggesting that SNORA42 overexpression is activated by its genomic amplification. SNORA42 knockdown in NSCLC cells inhibits in vitro and in vivo tumorigenicity, whereas enforced SNORA42 expression in bronchial epitheliums increases cell growth and colony formation. Such pleiotropy of SNORA42 suppression could be achieved at least partially through increased apoptosis of NSCLC cells in a p53-dependent manner. SNORA42 expression in lung tumor tissue specimens is inversely correlated with survival of NSCLC patients. Therefore, SNORA42 activation could have an oncogenic role in lung tumorigenesis and provide potential diagnostic and therapeutic targets for the malignancy.

Effects of fasting on hematologic and clinical chemical values in cynomolgus monkeys (Macaca fascicularis).

BACKGROUND: Fasting is an important pre-analytical factor that may affect clinical pathology parameters in toxicological and pharmacological studies. Little information is available on how fasting affects clinical pathology parameters in cynomolgus monkeys (Macaca fascicularis).The aim of this study was to evaluate the influence of fasting on clinical pathology parameters in healthy adult cynomolgus monkeys. METHODS: Five female and six male cynomolgus monkeys were fasted for 0, 8, 16, and 24 hours. Changes in body weight (BW), core hematologic, and serum clinical chemical parameters were evaluated. RESULTS: The BW significantly decreased after 24 hours of fasting. Significant decreases in red blood cell count, hemoglobin, hematocrit, and mean corpuscular volume and increases in mean cell hemoglobin and mean cell hemoglobin concentration were observed at 16 hours in males. In females, increasing the duration of fasting caused a significant time-dependent increase in platelets. Blood urea nitrogen showed significant decreases in female and male monkeys after fasting. Alkaline phosphatase increased in females after fasting. Aspartate transaminase significantly increased both in females and males at 8 hours. In females, alanine transaminase and lactate dehydrogenase significantly increased at 8 hours. Albumin significantly decreased in males 24 hours, but increased in females 16 hours after fasting. Serum glucose and triglyceride were not affected by fasting. Serum calcium decreased and inorganic phosphorus increased in males after fasting. CONCLUSION: These results suggested that clinical pathology data would vary after fasting. The decision to feed or fast before blood collection for clinical pathology tests should be made based on careful consideration.

Lupin alkaloids from Chinese Maackia amurensis.

Two new alkaloids were isolated together with 16 known lupin alkaloids from the leaves and stems of Chinese Maackia amurensis. Their structures were determined by spectroscopic methods to be (-)-6alpha-methoxylupanine and (-)-5alpha-(12-cytisinylmethyl)-6alpha-hydroxylupan ine and identified by comparison with synthetic samples. The structures of lupin alkaloids were also related to the geographical distributions of the Maackia plants.

Genotoxicity of drinking water from Chao Lake.

Genotoxic activity appears to originate primarily from reactions of chlorine with humic substances in the source waters. Comparisons of extracts of settled versus chlorinated water have confirmed that chlorinating during water treatment produces mutagenic activity in the mutagenicity tests. Present work on XAD-2 extracts of raw, chlorinated (treated), and settled water from the Chao Lake region of China has involved a battery of mutagenicity assays for various genetic endpoints: the Salmonella test, the sister-chromatid exchange (SCE) induction in Chinese hamster lung (CHL) cells, and the micronucleus (MN) induction in the peripheral blood erythrocytes of silver carp. Extracts of raw and treated water but not the settled water are mutagenic in the Salmonella assay. On the other hand, extracts of three water samples show activity in the SCE and MN assays, especially the raw and treated water. These data show that contamination and chlorinating contribute mutagens to drinking water and suggest that the mammalian assays may be more sensitive for detecting mutagenicity in aquatic environment than the Salmonella test.

[Variations of the essential oils in Acorua calamus L. and their major compositions].

The variations of the essential oils in Acorus calamus and their major compositions with seasons and producing areas were investigated by steam distillation and internal standard method. The result has shown that June is the best season for A. calamus cropping and that the major components of the volatile oil in the same part of the plant from different producing areas have the same chemical structure type.

[Studies on the constituents of the roots of Acorus calamus L].

A new tricyclic sesquiterpene, named calamenone (1), and two known sesquiterpenes, calamendiol (2) and isocalamendiol (3), were isolated from the roots of Acorus calamus L. The structure of the new compound was elucidated on the basis of spectral and X-ray diffraction data.

Effects of feeding flax and two types of sunflower seeds on fatty acid compositions of yolk lipid classes.

Single Comb White Leghorn laying hens were fed diets high in oleic, linolenic, or linoleic acid prepared by incorporation of high oleic acid sunflower seed (HOAS), full-fat flax seed (FLAX), or regular high linoleic acid sunflower seed (HLAS), respectively, to investigate the effects of dietary fats on the fatty acid compositions of major lipid classes of chicken eggs. Egg production, Haugh units, specific gravity, and yolk total lipid content were measured. After 3 wk of feeding, the fatty acid compositions of yolk total lipids, triglycerides, phosphatidylcholine (PC), and phosphatidylethanolamine (PE) were measured by gas chromatography. Dietary treatments had no effect on egg production, Haugh units, specific gravity, or yolk total lipid content. Feeding HOAS increased yolk oleic acid by 17%, and the change was only in triglycerides. The increases of yolk linoleic and arachidonic acids upon HLAS feeding were distributed evenly among triglycerides and PC, with moderate effect in the PE fraction. The enrichment of linolenic acid in eggs from the FLAX regime was mainly in triglycerides. The longer chain n-3 fatty acids, such as eicosapentaenoic, dososapentaenoic, and docosahexaenoic acids, were deposited exclusively in phospholipids, particularly in PE. The contents of the longer chain n-3 fatty acids in PE were three to seven times those in PC, indicating a preferential incorporation of these fatty acids into PE.

Molecular characterization of Saccharomyces cerevisiae URA6 gene. DNA sequence, mutagenesis analysis, and cell cycle regulation relevant to its suppression mechanism to cdc8 mutation.

Yeast SOC8 DNA fragment was isolated as a wild type dominant suppressor of cdc8 mutation. We have used Bal31 deletion analysis to define the minimal 1 kilobase HpaI-NcoI DNA element required for complementing the cdc8 mutation. The complementing sequence harbored a multicopy plasmid also enhanced by uridine monophosphate kinase in crude extracts. DNA sequence analysis revealed an open reading frame encoding a protein with a molecular weight of 24,949. Since our SOC8 sequence was identical to that of URA6 gene, which encodes uridine monophosphate kinase, we conclude that SOC8 is allelic with URA6, and we use the term URA6 hereafter. Northern blotting experiments showed that the size of mRNA is about 0.9 kibobases. Primer extension experiments showed multiple transcriptional starting sites primarily located at -160 The size and the deduced amino acid composition are consistent with information obtained from purified uridine monophosphate kinase. Thus, both molecular genetic and biochemical evidence supports a notion that the URA6 is SOC8 encoding a yeast uridine monophosphate kinase. Mutagenesis analysis of its putative nucleotide-binding site, altering essential lysine to glutamic acid, resulted in loss of its uridine monophosphate kinase activity. Complementation analysis studies indicated that the mutated ura6 gene abolished its ability to complement ura6 mutant cells; nor could it suppress cdc8 mutation. Unlike CDC8, the mRNA level of the URA6 gene did not fluctuate throughout the cell cycle; presumably, the temporal order of these two enzymatic activities might be different during cell cycle progression. These data may explain an incomplete suppression of cdc8 by URA6, as previously observed. Taken together, the results support our previous speculation that the suppression of the cdc8 mutation mechanisms by URA6 is due to the provision of the trans-acting dTMP kinase activity to complement the cdc8 defect.

Effect of feeding egg yolk powder on the plasma and tissue cholesterol levels in rats.

Two isonitrogenous diets, one containing 18.7% egg yolk powder (YP) and the other balanced with casein, lard, and cholesterol (CN), were fed to male Sprague-Dawley rats for 4 wk. Rats fed the YP diet had levels of plasma total cholesterol and high density lipoprotein cholesterol similar to those fed CN. Free and total cholesterol contents of liver, but not heart tissue, were significantly lower in YP-fed than in CN-fed rats.