Effect of processing on muscle structure and protein digestibility in vitro : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Food Technology at Massey University, Palmerston North, New Zealand

Figures 2-1, 2-2, 2-3, 2-4 and Table 2-2 are re-used with publishers' permission.The objective of this thesis was to investigate the effect of processing on meat protein properties, muscle structure and in vitro protein digestibility of beef. Meat processing techniques including pulsed electric field (PEF), shockwave (SW) processing, exogenous enzyme (actinidin) treatment, and sous vide (SV) cooking were explored, either alone or in combination, in this project. This thesis also aimed to study the diffusion of enzymes (actinidin from kiwifruit and pepsin in the gastric juice) into the meat. The first experiment investigated the effect of PEF processing alone on the ultrastructure and in vitro protein digestibility of bovine Longissimus thoracis, a tender meat cut (Chapter 3). It was observed that the moisture content of the PEF-treated samples (specific energy of 48 ± 5 kJ/kg and 178 ± 11 kJ/kg) was significantly lower (p < 0.05) by 1.3 to 4.6 %, compared to the untreated samples. The pH, colour, and protein thermal profile of the PEF-treated muscles remained unchanged. Pulsed electric field treatment caused the weakening of the Z-disk and I-band junctions and sarcomere elongation (25 to 38 % longer) of the muscles. The treatment improved in vitro meat protein digestibility by at least 18 %. In this thesis, the protein digestibility was determined in terms of the ninhydrin-reactive amino nitrogen released during simulated oral-gastro-small intestinal digestion. An enhanced proteolysis of the PEF-treated meat proteins (such as α-actinin and β-actinin subunit) during simulated digestion was also observed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The improvement in protein digestibility of the PEF-treated meat was supported by more severe disruption of Z-disks and I-bands observed in PEF-treated samples, at the end of simulated digestion. In the second experiment, PEF treatment (specific energy of 99 ± 5 kJ/kg) was applied to bovine Deep and Superficial pectoral muscles in conjunction with SV cooking (60 ℃ for 24 h) (Chapter 4). This muscle cut was tested as it is a tough cut and requires slow cooking. There was no significant difference detected in the specific activities of the sarcoplasmic cathepsins present in the cytosol between the control and PEF-treated samples, both before and after cooking. In addition, similar micro- and ultrastructures were observed between the control SV-cooked and PEF-treated SV-cooked pectoral muscles. The combined PEF-SV treatment increased the in vitro protein digestibility of the pectoral muscles by approximately 29 %. An improvement in proteolysis of the treated meat proteins (e.g. myosin heavy chains and C-protein) during simulated digestion was also observed using SDS-PAGE. More damaged muscle micro- and ultrastructures were detected in PEF-treated SV-cooked muscles at the end of in vitro oral-gastro-small intestinal digestion, showing its enhanced proteolysis compared to the control cooked meat. Next, the effect of SW processing and subsequent SV cooking on meat protein properties, muscle structure and in vitro protein digestibility of bovine Deep and Superficial pectoral muscles were investigated (Chapter 5 and 6). Shockwave processing (11 kJ/pulse) alone decreased the enthalpy and thermal denaturation temperature of the collagen (p < 0.05) when compared to the raw control, studied using a differential scanning calorimeter. The purge loss, pH, colour, and the protein gel electrophoresis profile of the SW-treated raw muscles remained unaffected. Shockwave processing led to the disorganisation of the sarcomere structure and also modified the protein secondary structure of the myofibres. After subsequent SV cooking (60 ℃ for 12 h), more severe muscle fibre coagulation and denaturation were observed in the SW-treated cooked meat compared to the cooked control. An increase in cook loss and a decrease in the Warner-Bratzler shear force were detected in the SW-treated SV-cooked meat compared to the control cooked meat (p < 0.05). The in vitro protein digestibility of the SW-treated SV-cooked meat was improved by approximately 22 %, with an enhanced proteolysis observed via SDS-PAGE, compared to the control SV-cooked meat. These results were supported by the observation of more destruction of the micro- and ultrastructures of SW-treated cooked muscles, observed at the end of the simulated digestion. The effect of the kiwifruit enzyme actinidin on muscle microstructure was studied using Picro-Sirius Red staining (Chapter 7). Meat samples were subjected to two different conditions, simulating meat marination (pH 5.6) and gastric digestion in humans (pH 3). Actinidin was found to have a greater proteolytic effect on the myofibrillar proteins than the connective tissue under both conditions. When compared with pepsin under simulated gastric conditions, actinidin had a weaker proteolytic effect on the connective tissue of cooked meats. Nevertheless, incubating the cooked meat in a solution containing both actinidin and pepsin resulted in more severe muscle structure degradation, when compared to muscles incubated in a single enzyme system. Thus, the co-ingestion of kiwifruit and meat could promote protein digestion of meat in the stomach. In addition, both actinidin and pepsin were successfully located at the edges of the muscle cells and in the endomysium using immunohistofluorescence imaging. The observations suggest that the incubation solutions penetrate into the muscle through the extracellular matrix to the intracellular matrix, enabling the proteases to access their substrates. Overall, the present work demonstrated that there were strong interactions between processing, muscle protein properties and structure, and in vitro protein digestibility of the meat. Processing induces changes in meat protein properties and muscle structure, which in turn affects the digestion characteristics of muscle-based foods

Tyrosinase Inhibitory Effect and Antioxidative Activities of Fermented and Ethanol Extracts of Rhodiola rosea

This is the first study to investigate the biological activities of fermented extracts of Rhodiola rosea L. (Crassulaceae) and Lonicera japonica Thunb. (Caprifoliaceae). Alcaligenes piechaudii CC-ESB2 fermented and ethanol extracts of Rhodiola rosea and Lonicera japonica were prepared and the antioxidative activities of different concentrations of samples were evaluated using in vitro antioxidative assays. Tyrosinase inhibition was determined by using the dopachrome method with L-DOPA as substrate. The results demonstrated that inhibitory effects (ED50 values) on mushroom tyrosinase of fermented Rhodiola rosea, fermented Lonicera japonica, ethanol extract of Lonicera japonica, and ethanol extract of Rhodiola rosea were 0.78, 4.07, 6.93, and >10 mg/ml, respectively. The DPPH scavenging effects of fermented Rhodiola rosea (ED50 = 0.073 mg/ml) and fermented Lonicera japonica (ED50 = 0.207 mg/ml) were stronger than effects of their respective ethanol extracts. Furthermore, the scavenging effect increases with the presence of high content of total phenol. However, the superoxide scavenging effects of fermented Rhodiola rosea was less than effects of fermented Lonicera japonica. The results indicated that fermentation of Rhodiola rosea and Lonicera japonica can be considered as an effective biochemical process for application in food, drug, and cosmetics

Active RIS-Assisted MIMO-OFDM System: Analyses and Prototype Measurements

In this study, we develop an active reconfigurable intelligent surface (RIS)-assisted multiple-input multiple-output orthogonal frequency division multiplexing (MIMO-OFDM) prototype compliant with the 5G New Radio standard at 3.5 GHz. The experimental results clearly indicate that active RIS plays a vital role in enhancing MIMO performance, surpassing passive RIS. Furthermore, when considering factors such as complexity, energy consumption, and performance, the comparative evaluation between passive RIS and active RIS reinforces the critical role of active RIS in MIMO systems. These findings underscore the practical significance of active RIS in improving MIMO gain in 5G scenarios.Comment: 5 pages, 6 figures, 1 table. This work has been submitted to the IEEE for possible publication. Copyright may be transferred without notice, after which this version may no longer be accessibl

Solar and Interplanetary Turbulence: Lagrangian Coherent Structures

Talk delivered in 22nd EGU General Assembly, held online 4-8 May, 2020, id.4289, https://meetingorganizer.copernicus.org/EGU2020/EGU2020-4289.html.-- https://www.egu2020.eu/The dynamics of solar and interplanetary plasmas is governed by coherent structures such as current sheets and magnetic flux ropes which are responsible for the genesis of intermittent turbulence via magnetic reconnections in solar supergranular junctions, solar coronal loops, the shock-sheath region of an interplanetary coronal mass ejection, and the interface region of two interplanetary magnetic flux ropes. Lagrangian coherent structures provide a new powerful technique to detect time- or space-dependent transport barriers, and objective (i.e., frame invariant) kinematic and magnetic vortices in space plasma turbulence. We discuss the basic concepts of Lagrangian coherent structures in plasmas based on the computation of the finite-time Lyapunov exponent, the Lagrangian averaged vorticity deviation and the integrated averaged current deviation, as well as their applications to numerical simulations of MHD turbulence and space and ground observations.With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation SEV-2017-070

IGRhCellID: integrated genomic resources of human cell lines for identification

Cell line identification is emerging as an essential method for every cell line user in research community to avoid using misidentified cell lines for experiments and publications. IGRhCellID (http://igrcid.ibms.sinica.edu.tw) is designed to integrate eight cell identification methods including seven methods (STR profile, gender, immunotypes, karyotype, isoenzyme profile, TP53 mutation and mutations of cancer genes) available in various public databases and our method of profiling genome alterations of human cell lines. With data validation of 11 small deleted genes in human cancer cell lines, profiles of genomic alterations further allow users to search for human cell lines with deleted gene to serve as indigenous knock-out cell model (such as SMAD4 in gene view), with amplified gene to be the cell models for testing therapeutic efficacy (such as ERBB2 in gene view) and with overlapped aberrant chromosomal loci for revealing common cancer genes (such as 9p21.3 homozygous deletion with co-deleted CDKN2A, CDKN2B and MTAP in chromosome view). IGRhCellID provides not only available methods for cell identification to help eradicating concerns of using misidentified cells but also designated genetic features of human cell lines for experiments

Probing Dark Matter Self-Interaction in the Sun with IceCube-PINGU

[[abstract]]We study the capture, annihilation and evaporation of dark matter (DM) inside the Sun. It has been shown that the DM self-interaction can increase the DM number inside the Sun. We demonstrate that this enhancement becomes more significant in the regime of small DM mass, given a fixed DM self-interaction cross section. This leads to the enhancement of neutrino flux from DM annihilation. On the other hand, for DM mass as low as as a few GeVs, not only the DM-nuclei scatterings can cause the DM evaporation, DM self-interaction also provides non-negligible contributions to this effect. Consequently, the critical mass for DM evaporation (typically 3 ~ 4 GeV without the DM self-interaction) can be slightly increased. We discuss the prospect of detecting DM self-interaction in IceCube-PINGU using the annihilation channels χχ → τ+τ-, νbar nu as examples. The PINGU sensitivities to DM self-interaction cross section σχχ are estimated for track and cascade events.[[notice]]補正完

Extra- and intra-ovarian factors in polycystic ovary syndrome: impact on oocyte maturation and embryo developmental competence

background: Polycystic ovary syndrome (PCOS) is a common metabolic dysfunction and heterogeneous endocrine disorder in women of reproductive age. Although patients with PCOS are typically characterized by increased numbers of oocytes retrieved during IVF, they are often of poor quality, leading to lower fertilization, cleavage and implantation rates, and a higher miscarriage rate. methods: For this review, we searched the database MEDLINE (1950 to January 2010) and Google for all full texts and/or abstract articles published in English with content related to oocyte maturation and embryo developmental competence. results: The search showed that alteration of many factors may directly or indirectly impair the competence of maturating oocytes through endocrine and local paracrine/autocrine actions, resulting in a lower pregnancy rate in patients with PCOS. The extra-ovarian factors identified included gonadotrophins, hyperandrogenemia and hyperinsulinemia, although intra-ovarian factors included members of the epidermal, fibroblast, insulin-like and neurotrophin families of growth factors, as well as the cytokines. conclusions: Any abnormality in the extra- and/or intra-ovarian factors may negatively affect the granulosa cell-oocyte interaction, oocyte maturation and potential embryonic developmental competence, contributing to unsuccessful outcomes for patients with PCOS who are undergoing assisted reproduction.Obstetrics &amp; GynecologyReproductive BiologySCI(E)PubMed49REVIEW117-331