Impact of calcium lactate and polysaccharides on skim milk gelation : a thesis presented in partial fulfilment of the requirements for the degree of Master in Food Technology at Massey University, Albany, New Zealand

Figures 2.1 (=Horne, 2006 Fig 1) & 2.6 (=Loren et al., 2007, Fig 9.3) were removed for copyright reasons.Swallowing difficulty is a growing health issue in adults particularly in the elderly population throughout the world. Thickeners and/or stabilisers are often added to the fluids to make them easier to swallow. Calcium-fortified dairy products have also been recommended for adults and the elderly to minimise the development of osteoporosis. Texture-modified milk with added calcium and polysaccharides could benefit adults who require an adequate calcium intake and have swallowing difficulties. Therefore, this project aimed to investigate the effect of the addition of calcium lactate to skim milk with and without polysaccharides. This project examined the calcium-added skim milk with and without polysaccharides for physical and rheological properties and microstructure. The first part of this project studied the effect of different heat treatments and the addition of different concentrations of calcium lactate on skim milk. The skim milk samples with added calcium lactate were heat treated at different temperatures (65°C, 70°C, 75°C and 80°C) and held at different periods of time (30 min or 60 min), followed by cooling to 20°C. The second part of this project investigated the effect of the addition of calcium lactate and three polysaccharides (xanthan gum, high acyl gellan gum and guar gum) on skim milk with calcium lactate after heating and holding at 75°C for 30 min. The physical properties of the skim milk samples were visually examined after heat treatment and the rheological properties were determined for different concentrations of polysaccharides using a rheometer. The microstructure of the samples was studied using a scanning electron microscope (SEM). Increasing the concentration of calcium lactate promoted the gelation of skim milk along with high heating temperatures (>70°C) and longer holding times. At the concentrations <10mM added calcium lactate, liquid skim milk samples with no gelation were observed. When the concentration of added calcium lactate was greater than 10mM, hard and firm gels were formed at all heat treatments. The final G’ increased with increasing concentration of added calcium lactate and higher heating temperature and longer holding times. The highest final G’ was achieved when the sample with 20mM added calcium lactate was heated at 80°C for 60 min. The addition of xanthan gum, high acyl gellan gum or guar gum demonstrated different interactions with calcium lactate in the milk system. At 10mM and 15mM calcium lactate, soft gels were formed when the xanthan concentration was less than 0.2% and viscous liquids were formed and became more viscous as the xanthan gum concentration increased above 0.2%. A dense and compact network with crosslinking was observed under SEM when 0.3% xanthan gum and 15mM calcium lactate were added to skim milk. With the addition of high acyl gellan gum, firmer gels were formed and the final G’ with increasing concentration of high acyl gellan gum, while a fibrous network was observed at 0.3% added high acyl gellan gum. With the addition of guar gum, soft gels were produced with aggregates and phase separation was observed when 0.2%-0.3% guar gum was added. The final G’ did not increase significantly (p<0.05) as the concentration of guar gum increased and a less compact structure was observed under SEM. The major findings in this study may be used as a guideline for the potential development of thickened milk beverages. The mouthfeel of the skim milk with 10mM and 15mM calcium lactate was improved when the xanthan concentration increased above 0.1% and a smooth mouthfeel was perceived at 0.5% added xanthan. Guar gum was not recommended to be used as undesirable mouthfeel was perceived

A 115-bp MethyLight assay for detection of p16 (CDKN2A) methylation as a diagnostic biomarker in human tissues

<p>Abstract</p> <p>Background</p> <p><it>p16 </it>Methylation is a potential biomarker for prediction of malignant transformation of epithelial dysplasia. A probe-based, quantitative, methylation-specific PCR (MSP) called MethyLight may become an eligible method for detecting this marker clinically. We studied oral mucosa biopsies with epithelial dysplasia from 78 patients enrolled in a published 4-years' followup cohort, in which cancer risk for patients with <it>p16 </it>methylation-positive dysplasia was significantly higher than those without <it>p16 </it>methylation (by 150-bp MSP and bisulfite sequencing; +133 ~ +283, transcription starting site, +1). The <it>p16 </it>methylation status in samples (<it>N </it>= 102) containing sufficient DNA was analyzed by the 70-bp classic (+238 ~ +307) and 115-bp novel (+157 ~ +272) MethyLight assays, respectively.</p> <p>Results</p> <p><it>p16 </it>Methylation was detectable in 75 samples using the classic MethyLight assay. The methylated-<it>p16 </it>positive rate and proportion of methylated-<it>p16 </it>by the MethyLight in MSP-positive samples were higher than those in MSP-negative samples (positive rate: 37/44 vs. 38/58, <it>P</it>=0.035, two-sided; proportion [median]: 0.78 vs. 0.02, <it>P <</it>0.007). Using the published results of MSP as a golden standard, we found sensitivity, specificity, and accuracy for this MethyLight assay to be 70.5%, 84.5%, and 55.0%, respectively. Because amplicon of the classic MethyLight procedure only partially overlapped with the MSP amplicon, we further designed a 115-bp novel MethyLight assay in which the amplicon on the sense-strand fully overlapped with the MSP amplicon on the antisense-strand. Using the 115-bp MethyLight assay, we observed methylated-<it>p16 </it>in 26 of 44 MSP-positive samples and 2 of 58 MSP-negative ones (<it>P </it>= 0.000). These results were confirmed with clone sequencing. Sensitivity, specificity, and accuracy using the 115-bp MethyLight assay were 59.1%, 98.3%, and 57.4%, respectively. Significant differences in the oral cancer rate were observed during the followup between patients (≥60 years) with and without methylated-<it>p16 </it>as detected by the 115-bp MethyLight assay (6/8 vs. 6/22, P = 0.034, two-sided).</p> <p>Conclusions</p> <p>The 115-bp MethyLight assay is a useful and practical assay with very high specificity for the detection of <it>p16 </it>methylation clinically.</p

A Chromosome-Level Genome Assembly of the Mandarin Fish (Siniperca chuatsi)

The mandarin fish, Siniperca chuatsi, is an economically important perciform species with widespread aquaculture practices in China. Its special feeding habit, acceptance of only live prey fishes, contributes to its delicious meat. However, little is currently known about related genetic mechanisms. Here, we performed whole-genome sequencing and assembled a 758.78 Mb genome assembly of the mandarin fish, with the scaffold and contig N50 values reaching 2.64 Mb and 46.11 Kb, respectively. Approximately 92.8% of the scaffolds were ordered onto 24 chromosomes (Chrs) with the assistance of a previously established genetic linkage map. The chromosome-level genome contained 19,904 protein-coding genes, of which 19,059 (95.75%) genes were functionally annotated. The special feeding behavior of mandarin fish could be attributable to the interaction of a variety of sense organs (such as vision, smell, and endocrine organs). Through comparative genomics analysis, some interesting results were found. For example, olfactory receptor (OR) genes (especially the beta and delta types) underwent a significant expansion, and endocrinology/vision related npy, spexin, and opsin genes presented various functional mutations. These may contribute to the special feeding habit of the mandarin fish by strengthening the olfactory and visual systems. Meanwhile, previously identified sex-related genes and quantitative trait locis (QTLs) were localized on the Chr14 and Chr17, respectively. 155 toxin proteins were predicted from mandarin fish genome. In summary, the high-quality genome assembly of the mandarin fish provides novel insights into the feeding habit of live prey and offers a valuable genetic resource for the quality improvement of this freshwater fish

Polycomb CBX7 Directly Controls Trimethylation of Histone H3 at Lysine 9 at the p16 Locus

BACKGROUND: H3K9 trimethylation (H3K9me3) and binding of PcG repressor complex-1 (PRC1) may play crucial roles in the epigenetic silencing of the p16 gene. However, the mechanism of the initiation of this trimethylation is unknown. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, we found that upregulating the expression of PRC1 component Cbx7 in gastric cancer cell lines MGC803 and BGC823 led to significantly suppress the expression of genes within the p16-Arf-p15 locus. H3K9me3 formation was observed at the p16 promoter and Regulatory Domain (RD). CBX7 and SUV39H2 binding to these regions were also detectable in the CBX7-stably upregulated cells. CBX7-SUV39H2 complexes were observed within nucleus in bimolecular fluorescence complementation assay (BiFC). Mutations of the chromodomain or deletion of Pc-box abolished the CBX7-binding and H3K9me3 formation, and thus partially repressed the function of CBX7. SiRNA-knockdown of Suv39h2 blocked the repressive effect of CBX7 on p16 transcription. Moreover, we found that expression of CBX7 in gastric carcinoma tissues with p16 methylation was significantly lower than that in their corresponding normal tissues, which showed a negative correlation with transcription of p16 in gastric mucosa. CONCLUSION/SIGNIFICANCE: These results demonstrated for the first time, to our knowledge, that CBX7 could initiate H3K9me3 formation at the p16 promoter

China’s 10-year progress in DC gas-insulated equipment: From basic research to industry perspective

The construction of the future energy structure of China under the 2050 carbon-neutral vision requires compact direct current (DC) gas-insulation equipment as important nodes and solutions to support electric power transmission and distribution of long-distance and large-capacity. This paper reviews China's 10-year progress in DC gas-insulated equipment. Important progresses in basic research and industry perspective are presented, with related scientific issues and technical bottlenecks being discussed. The progress in DC gas-insulated equipment worldwide (Europe, Japan, America) is also reported briefly

The Effect of Knocked-Down Anti-M&uuml;llerian Hormone mRNA on Reproductive Characters of Male Nile Tilapia (Oreochromis niloticus) through Inhibition of the TGF-Beta Signaling Pathway

Anti-M&uuml;llerian hormone (amh), an important regulator of gonad development in male teleosts, regulates the development and differentiation of germ cells. We performed transcriptional knock-down of amh in Nile tilapia (Oreochromis niloticus) using antisense RNA technology, resulting in down-regulation in the expression of amh transcription and Amh protein in males. Compared with the control groups, the fish in treatment groups with down-regulated amh had increased weight and an extremely significant decrease in the gonadosomatic index. Hematoxylin&ndash;eosin staining revealed impaired testis development and significant reductions in numbers of sperm. Serum estradiol levels were significantly increased, and the levels of testosterone, luteinizing hormone, and follicle-stimulating hormone were significantly decreased. RNA-sequencing analysis of the fish in the down-regulated amh and control groups identified 12,048 differentially expressed genes, of which 1281 were up-regulated and 10,767 were down-regulated. Kyoto Encyclopedia of Genes and Genomes analysis revealed that differentially expressed genes related to growth and development were mainly enriched in the Cell cycle, Endocytosis, TGF-beta signaling pathway, Wnt signaling pathway, FoxO signaling pathway, Insulin signaling pathway, and MAPK signaling pathway. The RNA-sequencing data accuracy was verified by qRT-PCR analysis of the expression levels of selected differentially expressed genes. The abnormal TGF-beta signaling pathway may cause fish weight gain, testis dysplasia, and abnormal spermatogenesis: smad5, smad3a, tgfb2, tgfbr1b, gsdf, and amh were significantly down-regulated. These findings indicated that antisense RNA technology has strong application prospects and can specifically knock down amh in Nile tilapia, resulting in an abnormal TGF-beta signaling pathway, inhibiting testis development and inducing weight gain

The Effect of Knocked-Down Anti-Müllerian Hormone mRNA on Reproductive Characters of Male Nile Tilapia (<i>Oreochromis niloticus</i>) through Inhibition of the TGF-Beta Signaling Pathway

Anti-Müllerian hormone (amh), an important regulator of gonad development in male teleosts, regulates the development and differentiation of germ cells. We performed transcriptional knock-down of amh in Nile tilapia (Oreochromis niloticus) using antisense RNA technology, resulting in down-regulation in the expression of amh transcription and Amh protein in males. Compared with the control groups, the fish in treatment groups with down-regulated amh had increased weight and an extremely significant decrease in the gonadosomatic index. Hematoxylin–eosin staining revealed impaired testis development and significant reductions in numbers of sperm. Serum estradiol levels were significantly increased, and the levels of testosterone, luteinizing hormone, and follicle-stimulating hormone were significantly decreased. RNA-sequencing analysis of the fish in the down-regulated amh and control groups identified 12,048 differentially expressed genes, of which 1281 were up-regulated and 10,767 were down-regulated. Kyoto Encyclopedia of Genes and Genomes analysis revealed that differentially expressed genes related to growth and development were mainly enriched in the Cell cycle, Endocytosis, TGF-beta signaling pathway, Wnt signaling pathway, FoxO signaling pathway, Insulin signaling pathway, and MAPK signaling pathway. The RNA-sequencing data accuracy was verified by qRT-PCR analysis of the expression levels of selected differentially expressed genes. The abnormal TGF-beta signaling pathway may cause fish weight gain, testis dysplasia, and abnormal spermatogenesis: smad5, smad3a, tgfb2, tgfbr1b, gsdf, and amh were significantly down-regulated. These findings indicated that antisense RNA technology has strong application prospects and can specifically knock down amh in Nile tilapia, resulting in an abnormal TGF-beta signaling pathway, inhibiting testis development and inducing weight gain