LC-ESI-QTOF-MS/MS Identification and Characterization of Phenolic Compounds from Leaves of Australian Myrtles and Their Antioxidant Activities.

Phenolic compounds, present in plants, provide substantial health advantages, such as antioxidant and anti-inflammatory properties, which enhance cardiovascular and cognitive well-being. Australia is enriched with a wide range of plants with phytopharmacological potential, which needs to be fully elucidated. In this context, we analyzed leaves of aniseed myrtle (Syzygium anisatum), lemon myrtle (Backhousia citriodora), and cinnamon myrtle (Backhousia myrtifolia) for their complex phytochemical profile and antioxidant potential. LC-ESI-QTOF-MS/MS was applied for screening and characterizing these Australian myrtles' phenolic compounds and the structure-function relation of phenolic compounds. This study identified 145 and quantified/semi-quantified 27 phenolic compounds in these Australian myrtles. Furthermore, phenolic contents (total phenolic content (TPC), total condensed tannins (TCT), and total flavonoids (TFC)) and antioxidant potential of phenolic extracts from the leaves of Australian myrtles were quantified. Aniseed myrtle was quantified with the highest TPC (52.49 ± 3.55 mg GAE/g) and total antioxidant potential than other selected myrtles. Catechin, epicatechin, isovitexin, cinnamic acid, and quercetin were quantified as Australian myrtles' most abundant phenolic compounds. Moreover, chemometric analysis further validated the results. This study provides a new insight into the novel potent bioactive phenolic compounds from Australian myrtles that could be potentially useful for functional, nutraceutical, and therapeutic applications.

Screening and Characterization of Phenolic Compounds from Selected Unripe Fruits and Their Antioxidant Potential.

The food sector's interest in sustainability and the demand for novel bioactive compounds are increasing. Many fruits are wasted every year before ripening due to various climatic conditions and harsh weather. Unripe mangoes, grapes, and black lemons could be rich sources of phenolic compounds that need to be fully elucidated. Using fruit waste as a source of bioactive chemicals has grown increasingly appealing as it may have significant economic benefits. Polyphenols are beneficial for human health to inhibit or minimize oxidative stress and can be used to develop functional and nutraceutical food products. In this context, this study aimed to characterize and screen unripe mangoes, grapes, and black lemons for phenolic compounds using LC-ESI-QTOF-MS/MS and their antioxidant activities. Unripe mangoes were quantified with higher total phenolic content (TPC, 58.01 ± 6.37 mg GAE/g) compared to black lemon (23.08 ± 2.28 mg GAE/g) and unripe grapes (19.42 ± 1.16 mg GAE/g). Furthermore, unripe mangoes were also measured with higher antioxidant potential than unripe grapes and black lemons. A total of 85 phenolic compounds (70 in black lemons, 49 in unripe grapes, and 68 in unripe mango) were identified, and 23 phenolic compounds were quantified using LC-MS/MS. Procyanidin B2, gallic acid, epicatechin, caffeic acid, quercetin, and chlorogenic acid were measured with higher concentration in these selected unripe fruits. A positive correlation was found between phenolic contents and the antioxidant activities of unripe fruits. Furthermore, chemometric analysis was conducted to validate the results. This study will explore the utilization of these unripe fruits to develop functional and therapeutic foods.

A Comparative Study for Nutritional and Phytochemical Profiling of Coffea arabica (C. arabica) from Different Origins and Their Antioxidant Potential and Molecular Docking.

Coffee is the most widely used beverage globally and contains many bioactive compounds, including phenolic compounds, alkaloids, triterpenes, organic acids, amino acids, hormones, and fatty acids. The main objective of this study was the comparative profiling of Australian, Colombian, Ethiopian, and Peruvian C. arabica using LC-ESI-QTOF-MS/MS. In this study, we tentatively identified 136 bioactive metabolites, including five (05) organic acids, six (06) alkaloids, three (03) amino acids (l-phenylalanine, l-tyrosine, and l-pyroglutamic acid), two (02) hormones (melatonin and serotonin), two fatty acids, one (01) furopyrans (goniothalenol), one (01) carotenoid (crocetin), three (03) terpenoids, thirty-eight (38) phenolic acids, forty-one (41) flavonoids, five (05) stilbenes, three (03) lignans and twenty-three (23) other polyphenols in C. arabica. The highest TPC value (17.74 ± 0.32 mg GAE/g) was measured in Colombian coffee while the lowest TPC value (10.24 ± 0.73 mg GAE/g) was in Peruvian coffee. Colombian coffee has a higher antioxidant potential than other studied coffee samples. A total of nineteen phenolic metabolites were mapped through LC-MS/MS. Quinic acid derivatives were quantified in higher concentrations than other metabolites. Furthermore, molecular docking predicted that chlorogenic acid is a main bioactive compound that contributes to anti-Alzheimer and anti-diabetic activities of C. arabica. The obtained results indicate that C. arabica contains a vast number of bioactive compounds which have potential health benefits. Furthermore, research could be conducted to validate the effect of these metabolites on the flavor profile of coffee beverages.

Digital technologies to assess yoghurt quality traits and consumers acceptability.

BACKGROUND: Sensory biometrics provide advantages for consumer tasting by quantifying physiological changes and the emotional response from participants, removing variability associated with self-reported responses. The present study aimed to measure consumers' emotional and physiological responses towards different commercial yoghurts, including dairy and plant-based yoghurts. The physiochemical properties of these products were also measured and linked with consumer responses. RESULTS: Six samples (Control, Coconut, Soy, Berry, Cookies and Drinkable) were evaluated for overall liking by n = 62 consumers using a nine-point hedonic scale. Videos from participants were recorded using the Bio-Sensory application during tasting to assess emotions and heart rate. Physicochemical parameters Brix, pH, density, color (L, a and b), firmness and near-infrared (NIR) spectroscopy were also measured. Principal component analysis and a correlation matrix were used to assess relationships between the measured parameters. Heart rate was positively related to firmness, yaw head movement and overall liking, which were further associated with the Cookies sample. Two machine learning regression models were developed using (i) NIR absorbance values as inputs to predict the physicochemical parameters (Model 1) and (ii) the outputs from Model 1 as inputs to predict consumers overall liking (Model 2). Both models presented very high accuracy (Model 1: R = 0.98; Model 2: R = 0.99). CONCLUSION: The presented methods were shown to be highly accurate and reliable with respect to their potential use by the industry to assess yoghurt quality traits and acceptability. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Maternal Heat Stress Alters Expression of Genes Associated with Nutrient Transport Activity and Metabolism in Female Placentae from Mid-Gestating Pigs.

Placental insufficiency is a known consequence of maternal heat stress during gestation in farm animals. The molecular regulation of placentae during the stress response is little known in pigs. This study aims to identify differential gene expression in pig placentae caused by maternal heat exposure during early to mid-gestation. RNA sequencing (RNA-seq) was performed on female placental samples from pregnant pigs exposed to thermoneutral control (CON; constant 20 °C; n = 5) or cyclic heat stress (HS; cyclic 28 to 33 °C; n = 5) conditions between d40 and d60 of gestation. On d60 of gestation, placental efficiency (fetal/placental weight) was decreased (p = 0.023) by maternal HS. A total of 169 genes were differentially expressed (FDR ≤ 0.1) between CON and HS placentae of female fetuses, of which 35 genes were upregulated and 134 genes were downregulated by maternal HS. The current data revealed transport activity (FDR = 0.027), glycoprotein biosynthetic process (FDR = 0.044), and carbohydrate metabolic process (FDR = 0.049) among the terms enriched by the downregulated genes (HS vs. CON). In addition, solute carrier (SLC)-mediated transmembrane transport (FDR = 0.008) and glycosaminoglycan biosynthesis (FDR = 0.027), which modulates placental stroma synthesis, were identified among the pathways enriched by the downregulated genes. These findings provide evidence that heat-stress induced placental inefficiency may be underpinned by altered expression of genes associated with placental nutrient transport capacity and metabolism. A further understanding of the molecular mechanism contributes to the identification of placental gene signatures of summer infertility in pigs.

Effects of L-citrulline supplementation on heat stress physiology, lactation performance and subsequent reproductive performance of sows in summer.

Lactating sows are susceptible to heat stress (HS). Part of the thermoregulatory response to HS is to increase peripheral blood flow, which is mediated in part by the vasodilator, nitric oxide (NO). Therefore, the aim of this experiment was to determine the effect of supplementation of L-citrulline, a NO precursor, on symptoms of HS, lactation performance and subsequent reproductive performance of sows in summer. A total of 221 summer farrowing mixed parity sows were fed either a control diet or supplemented with 1% L-citrulline upon entry to the farrowing house (6 ± 1.8 days for mean ± standard deviation [SD] before farrowing) until weaning (26 ± 1.5 days). The average daily minimum and maximum temperature in the farrowing house was 21.0 ± 1.88 and 29.2 ± 3.82°C (mean ± SD). Rectal temperature, respiration rate, and plasma and urinary nitrite and nitrate (NOx) of sows were measured on the 19th day post-farrowing. Supplemental L-citrulline in the diet did not affect the number of piglets born alive, feed intake of sows, body weight or backfat thickness of sows at weaning, or litter weight gain. L-citrulline tended to reduce piglet pre-weaning mortality rate from 18.6% to 15.6% (p = 0.058). L-citrulline reduced the respiration rate of sows compared to the control diet at 17:00 hr (Time × Diet, p < 0.001); however, rectal temperature was not affected. L-citrulline tended to increase urinary NOx concentrations (127 vs. 224 µM, p = 0.057) but not plasma NOx concentrations. L-citrulline did not affect farrowing rate or number of piglets born alive in the subsequent parity. In conclusion, L-citrulline supplementation reduced respiration rate of lactating sows and reduced piglet pre-weaning mortality rate in summer. Whether the effects were due to a NO-dependent mechanism requires further validation.

Effect of feeding slowly fermentable grains on productive variables and amelioration of heat stress in lactating dairy cows in a sub-tropical summer.

Feeding low-fiber and high-energy diets to dairy cows is one approach to ameliorate heat stress (HS) by reducing heat increment (HI) during digestion. However, rapidly and slowly fermentable cereal grains differ in their HI. The aim of this experiment was to quantify if feeding slowly fermentable grains ameliorated the physiological responses to HS and improved milk production (MP) in dairy cows. Holstein-Friesian lactating dairy cows were housed in shaded pens and were fed either a total mixed ration (TMR) plus wheat (TMRW), a TMR plus wheat treated with 2% of a commercial starch-binding agent (TMRB), or a TMR plus corn (TMRC) (n = 8 cows per diet) during summer in Queensland, Australia. Respiration rate (RR) and panting score (PS) were measured four times a day; rumen temperature (RuT) was recorded every 20 min, and rectal temperature (RT) and milk samples were obtained every 4 days. Cows fed slowly fermentable grains had higher milk production (MP) than cows fed TMRW, and cows fed TMRC had lower RT than those fed TMRW and TMRB (P < 0.001). Rumen temperature was positively correlated with temperature-humidity index and negatively correlated with MP (P < 0.05). In summary, feeding TMRC ameliorated HS as indicated by lower RT and improved MP in dairy cows. Milk production was improved with starch-binding agents; however, this was not associated with efficient thermoregulatory responses. Furthermore, determination of RuT enabled the prediction of changes in physiological variables and productive responses due to HS in lactating dairy cows.

A short-term supranutritional vitamin E supplementation alleviated respiratory alkalosis but did not reduce oxidative stress in heat stressed pigs.

OBJECTIVE: Heat stress (HS) triggers oxidative stress and respiratory alkalosis in pigs. The objective of this experiment was to study whether a short-term supranutritional amount of dietary vitamin E (VE) can mitigate oxidative stress and respiratory alkalosis in heat-stressed pigs. METHODS: A total of 24 pigs were given either a control diet (17 IU/kg VE) or a high VE (200 IU/kg VE; HiVE) diet for 14 d, then exposed to thermoneutral (TN; 20°C, 45% humidity) or HS (35°C, 35% to 45% humidity, 8 h daily) conditions for 7 d. Respiration rate and rectal temperature were measured three times daily during the thermal exposure. Blood gas variables and oxidative stress markers were studied in blood samples collected on d 7. RESULTS: Although HiVE diet did not affect the elevated rectal temperature or respiration rate observed during HS, it alleviated (all p<0.05 for diet×temperature) the loss of blood CO2 partial pressure and bicarbonate, as well as the increase in blood pH in the heat-stressed pigs. The HS reduced (p = 0.003) plasma biological antioxidant potential (BAP) and tended to increase (p = 0.067) advanced oxidized protein products (AOPP) in the heat-stressed pigs, suggesting HS triggers oxidative stress. The HiVE diet did not affect plasma BAP or AOPP. Only under TN conditions the HiVE diet reduced the plasma reactive oxygen metabolites (p<0.05 for diet× temperature). CONCLUSION: A short-term supplementation with 200 IU/kg VE partially alleviated respiratory alkalosis but did not reduce oxidative stress in heat-stressed pigs.

Supplementation of selenium, vitamin E, chromium and betaine above recommended levels improves lactating performance of sows over summer.

Heat stress (HS) exacerbates the body weight loss of lactating sows and reduces litter weight gain. Selenium (Se), vitamin E (VE), chromium (Cr) and betaine have been shown to ameliorate symptoms of HS, and yeast nucleotides and mannan oligosaccharides have been reported to improve lactational performance and immune response in pigs. Therefore, a combination of these nutrients may improve lactational performance of sows in summer. The effects of two nutritionally enhanced diets on lactational performance of sows in summer were investigated in two experiments. In experiment 1, we compared the effects of a nutritionally fortified diet (0.4 ppm Se, 95 IU/kg VE, 0.4 ppm Cr and 0.2% betaine; named as SVCB diet) with the NRC 2012 standard diet (0.15 ppm Se, 44 IU/kg VE) on lactational performance of sows in summer. Results showed that the SVCB diet reduced body weight loss (P = 0.039) and tended to reduce backfat loss (P = 0.075) of sows without affecting feed intake, while litter weight gain was not influenced. In experiment 2, we further enhanced the nutrients in the SVCB diet (0.8 ppm Se, 1% yeast nucleotides, and 0.1% mannan oligosaccharides; named as SNM diet). Results showed that the SNM diet did not improve feed intake of sows, farrowing performance, or litter weight gain compared with the SVCB diet, but increased body weight loss of the third parity sows (P = 0.037). Overall, a combined supplementation of Se, VE, Cr, and betaine above the NRC recommended levels can reduce mobilisation of body reserve of lactating sows in summer.

Selenium and vitamin E together improve intestinal epithelial barrier function and alleviate oxidative stress in heat-stressed pigs.

What is the central question of this study? Oxidative stress may play a role in compromising intestinal epithelial barrier integrity in pigs subjected to heat stress, but it is unknown whether an increase of dietary antioxidants (selenium and vitamin E) could alleviate gut leakiness in heat-stressed pigs. What is the main finding and its importance? Levels of dietary selenium (1.0 p.p.m.) and vitamin E (200 IU kg(-1) ) greater than those usually recommended for pigs reduced intestinal leakiness caused by heat stress. This finding suggests that oxidative stress plays a role in compromising intestinal epithelial barrier integrity in heat-stressed pigs and also provides a nutritional strategy for mitigating these effects. Heat stress compromises the intestinal epithelial barrier integrity of mammals through mechanisms that may include oxidative stress. Our objective was to test whether dietary supplementation with antioxidants, selenium (Se) and vitamin E (VE), protects intestinal epithelial barrier integrity in heat-stressed pigs. Female growing pigs (n = 48) were randomly assigned to four diets containing from 0.2 p.p.m. Se and 17 IU kg(-1) VE (control, National Research Council recommended) to 1.0 p.p.m. Se and 200 IU kg(-1) VE for 14 days. Six pigs from each dietary treatment were then exposed to either thermoneutral (20°C) or heat-stress conditions (35°C 09.00-17.00 h and 28°C overnight) for 2 days. Transepithelial electrical resistance and fluorescein isothiocyanate-dextran (4 kDa; FD4) permeability were measured in isolated jejunum and ileum using Ussing chambers. Rectal temperature, respiratory rate and intestinal HSP70 mRNA abundance increased (all P < 0.001), and respiratory alkalosis occurred, suggesting that pigs were heat stressed. Heat stress also increased FD4 permeability and decreased transepithelial electrical resistance (both P < 0.01). These changes were associated with changes indicative of oxidative stress, a decreased glutathione peroxidase (GPX) activity and an increased glutathione disulfide (GSSG)-to-glutathione (GSH) ratio (both P < 0.05). With increasing dosage of Se and VE, GPX-2 mRNA (P = 0.003) and GPX activity (P = 0.049) increased linearly, the GSSG:GSH ratio decreased linearly (P = 0.037), and the impacts of heat stress on intestinal barrier function were reduced (P < 0.05 for both transepithelial electrical resistance and FD4 permeability). In conclusion, in pigs an increase of dietary Se and VE mitigated the impacts of heat stress on intestinal barrier integrity, associated with a reduction in oxidative stress.

Milk-derived ribonuclease 5 preparations induce myogenic differentiation in vitro and muscle growth in vivo.

Ribonuclease 5, also known as angiogenin, is a stable and abundant ribonuclease in milk whey protein, which is able to regulate several cellular functions, including capillary formation, neuron survival, and epithelial cell growth. Ribonuclease 5 is important for protein synthesis directly stimulating rRNA synthesis in the nucleolus. Here, we show that biologically active RNase5 can be purified from bovine milk. Furthermore, we show that milk-derived RNase5 directly stimulates muscle cell differentiation in vitro, inducing C2C12 cell differentiation and myogenesis. When supplemented into the diet of healthy adult mice, milk-derived RNase5 preparations promoted muscle weight gain and grip strength. Collectively, these data indicate that milk-derived RNase5 preparations exhibit a novel role in skeletal muscle cell function.

Effects of Sugarcane-Derived Polyphenol Supplementation on Methane Production and Rumen Microbial Diversity of Second-Cross Lambs.

The objective of this study was to evaluate the effects of feeding sugarcane-derived polyphenolic supplement (Polygain, The Product Makers Australia, Keysborough, VIC, Australia) on enteric methane (CH4) emission, rumen microbiota, and performance of second-cross lambs. For this purpose, 24 Poll Dorset × (Border Leicester × Merino) lambs were allocated to 3 different treatments: Control (C), 0.25% Polygain (0.25 PG), and 1% Polygain (1 PG) diets with a uniform basal feed (25% cracked wheat grain, 25% cracked barley grain, 25% oaten chaff, 25% lucerne chaff). Both doses of Polygain reduced the total CH4 production (g/day; p = 0.006), CH4 yield (CH4, g/kg of dry matter intake; p = 0.003) and CH4 intensity (CH4, g/kg of BW; p = 0.003). Dry matter intake tended to be greater (p = 0.08) in sheep fed 1 PG compared to the C group, with the 0.25 PG group being intermediate. The average daily gain of the lambs was improved (p = 0.03) with 1% Polygain supplementation. The relative abundance of genera Methanobrevibacter_unidentified, Methanomethylophilaceae_uncultured, Methanogenic archaeon mixed culture ISO4-G1, Methanosphaera uncultured rumen methanogen, Methanogenic archaeon ISO4-H5, and Methanobrevibacter boviskoreani JH1 were reduced with Polygain supplementation. In conclusion, feeding Polygain reduced lambs' enteric CH4 emissions, altered the rumen microbiome, and improved the growth performance of lambs.

A comparative analysis of gastrointestinal tract barrier function and immune markers in gilt vs. sow progeny at birth and weaning.

Progeny born to primiparous sows (gilt progeny; GP) have lower birth, weaning and slaughter weights than sow progeny (SP). GP also have reduced gastrointestinal tract (GIT) development, as evidenced by lower organ weights. Therefore, the aim of this experiment was to quantify changes in GIT barrier function that occur in birth and weaning, representing two major challenges to the young piglet. The effects of parity (GP vs. SP) in GIT barrier integrity function were quantified at four timepoints: birth (~0 h), 24 h after birth (24 h), 1-d preweaning (PrW), and 1-d postweaning (PoW) in commercially reared piglets. Due to inherent differences between newborn and weanling pigs, the results were analyzed in two cohorts, birth (0 vs. 24 h, n = 31) and weaning (PrW vs. PoW, n = 40). Samples of the stomach, jejunum, ileum, and colon were excised after euthanasia and barrier integrity was quantified by measuring transepithelial resistance (TER), macromolecular permeability, the abundance of inflammatory proteins (IL-8, IL-1ß, and TNF-α) and tight junction proteins (claudin-2 and -3). Papp was characterized using a dual tracer approach comprising 4 KDa fluorescein isothiocyanate (FD4) and 150 kDa tetramethyl rhodamine isothiocyanate (T150)-labeled dextrans. Characteristic effects of the initiation of feeding and weaning were observed on the GIT with the initiation of feeding, such as increasing TER and reducing Papp at 24 h, consistent with mucosal growth (P = 0.058) This was accompanied by increased cytokine abundance as evidenced by elevations in TNF-α and IL-1ß. However, GP had increased IL-8 abundance (P = 0.011 and 0.063 for jejunum and ileum respectively) at birth than 24 h overall. In the weaning cohort, jejunal and ileal permeability to FD4 was higher in GP (P = 0.05 and 0.022, respectively) while only higher ileal T150 was observed in GP (P = 0.032). Ileal claudin-2 abundance tended to be higher in SP overall (P = 0.063), but GP ileal claudin-2 expression was upregulated weaning while no change was observed in SP (P = 0.043). Finally, other than a higher jejunal TNF-α abundance observed in SP (P = 0.016), no other effect of parity was observed on inflammatory markers in the weaning cohort. The results from this study indicate that the GIT of GP have poorer adaptation to early life events, with the response to weaning, being more challenging which is likely to contribute to poorer postweaning growth.

The progeny of primiparous sows (gilt progeny; GP) have poorer lifetime growth performance in comparison to progeny from multiparous sows (sow progeny; SP). Previous research suggests that there is an underlying biological basis for reduced growth performance which is attributed to differences in gastrointestinal tract (GIT) barrier development during early life. This study aimed to clarify the timeframes of when these differences are in effect by investigating GIT development during two major events of a piglet's life: birth and weaning. To do this, GIT tissue was collected from GP and SP at four time points; birth, 24 h after birth, 1-d preweaning, and 1-d postweaning and assessed for functional development. The main findings from this study indicate there are early signs of variation in GIT development within the first 24 h of life between GP and SP, and that these differences increase through the preweaning period, with GP entering weaning with poorer GIT development and function. Possible explanations for the reduced GI development may be reduced maternal nutrition during the suckling period.

Oral ivermectin versus malathion lotion for difficult-to-treat head lice.

BACKGROUND: Head-lice infestation is prevalent worldwide, especially in children 3 to 11 years old. Topical insecticides (i.e., pyrethroids and malathion) used as a lotion, applied twice at an interval of 7 to 11 days, are typically used for treatment. Resistance of lice to insecticides, particularly pyrethroids, results in treatment failure. The efficacy of alternative agents is controversial. METHODS: We conducted a multicenter, cluster-randomized, double-blind, double-dummy, controlled trial comparing oral ivermectin (at a dose of 400 microg per kilogram of body weight) with 0.5% malathion lotion, each given on days 1 and 8, for patients with live lice not eradicated by topical insecticide used 2 to 6 weeks before enrollment. The cluster was defined as the household. Infestation was confirmed and monitored by means of fine-toothed combing. Patients were at least 2 years of age and weighed at least 15 kg; all were treated at the study sites. The primary end point was the absence of head lice on day 15. RESULTS: A total of 812 patients from 376 households were randomly assigned to receive either ivermectin or malathion. In the intention-to-treat population, 95.2% of patients receiving ivermectin were lice-free on day 15, as compared with 85.0% of those receiving malathion (absolute difference, 10.2 percentage points; 95% confidence interval [CI], 4.6 to 15.7; P<0.001). In the per-protocol population, 97.1% of patients in the ivermectin group were lice-free on day 15, as compared with 89.8% of those in the malathion group (absolute difference, 7.3 percentage points; 95% CI, 2.8 to 11.8; P=0.002). There were no significant differences in the frequencies of adverse events between the two treatment groups. CONCLUSIONS: For difficult-to-treat head-lice infestation, oral ivermectin, given twice at a 7-day interval, had superior efficacy as compared with topical 0.5% malathion lotion, a finding that suggests that it could be an alternative treatment. (ClinicalTrials.gov number, NCT00819520.)

Genes of the RNASE5 pathway contain SNP associated with milk production traits in dairy cattle.

BACKGROUND: Identification of the processes and mutations responsible for the large genetic variation in milk production among dairy cattle has proved challenging. One approach is to identify a biological process potentially involved in milk production and to determine the genetic influence of all the genes included in the process or pathway. Angiogenin encoded by angiogenin, ribonuclease, RNase A family 5 (RNASE5) is relatively abundant in milk, and has been shown to regulate protein synthesis and act as a growth factor in epithelial cells in vitro. However, little is known about the role of angiogenin in the mammary gland or if the polymorphisms present in the bovine RNASE5 gene are associated with lactation and milk production traits in dairy cattle. Given the high economic value of increased protein in milk, we have tested the hypothesis that RNASE5 or genes in the RNASE5 pathway are associated with milk production traits. First, we constructed a "RNASE5 pathway" based on upstream and downstream interacting genes reported in the literature. We then tested SNP in close proximity to the genes of this pathway for association with milk production traits in a large dairy cattle dataset. RESULTS: The constructed RNASE5 pathway consisted of 11 genes. Association analysis between SNP in 1 Mb regions surrounding these genes and milk production traits revealed that more SNP than expected by chance were associated with milk protein percent (P < 0.05 significance). There was no significant association with other traits such as milk fat content or fertility. CONCLUSIONS: These results support a role for the RNASE5 pathway in milk production, specifically milk protein percent, and indicate that polymorphisms in or near these genes explain a proportion of the variation for this trait. This method provides a novel way of understanding the underlying biology of lactation with implications for milk production and can be applied to any pathway or gene set to test whether they are responsible for the variation of complex traits.

Antioxidant, Alpha-Glucosidase Inhibition Activities, In Silico Molecular Docking and Pharmacokinetics Study of Phenolic Compounds from Native Australian Fruits and Spices.

Native Australian fruits and spices are enriched with beneficial phytochemicals, especially phenolic compounds, which are not fully elucidated. Therefore, this study aimed to analyze native Australian mountain-pepper berries (Tasmannia lanceolata), rosella (Hibiscus sabdariffa), lemon aspen (Acronychia acidula), and strawberry gum (Eucalyptus olida) for phenolic and non-phenolic metabolites and their antioxidant and alpha-glucosidase inhibition activities. Liquid chromatography-mass spectrometry-electrospray ionization coupled with quadrupole time of flight (LC-ESI-QTOF-MS/MS) was applied to elucidate the composition, identities, and quantities of bioactive phenolic metabolites in Australian native commercial fruits and spices. This study identified 143 phenolic compounds, including 31 phenolic acids, 70 flavonoids, 10 isoflavonoids, 7 tannins, 3 stilbenes, 7 lignans, 10 other compounds, and 5 limonoids. Strawberry gum was found to have the highest total phenolic content (TPC-36.57 ± 1.34 milligram gallic acid equivalent per gram (mg GAE/g), whereas lemon aspen contained the least TPC (4.40 ± 0.38 mg GAE/g). Moreover, strawberry gum and mountain pepper berries were found to have the highest antioxidant and anti-diabetic potential. In silico molecular docking and pharmacokinetics screening were also conducted to predict the potential of the most abundant phenolic compounds in these selected plants. A positive correlation was observed between phenolic contents and biological activities. This study will encourage further research to identify the nutraceutical and phytopharmaceutical potential of these native Australian fruits.

Characterization, Antioxidant Potential, and Pharmacokinetics Properties of Phenolic Compounds from Native Australian Herbs and Fruits.

In recent decades, plant bioactive phenolic compounds gained much attention due to their various health benefits. Therefore, this study aimed to analyze native Australian river mint (Mentha australis), bush mint (Mentha satureioides), sea parsley (Apium prostratum), and bush tomatoes (Solanum centrale) for their bioactive metabolites, antioxidant potential, and pharmacokinetics properties. LC-ESI-QTOF-MS/MS was applied to elucidate these plants' composition, identification, and quantification of phenolic metabolites. This study tentatively identified 123 phenolic compounds (thirty-five phenolic acids, sixty-seven flavonoids, seven lignans, three stilbenes, and eleven other compounds). Bush mint was identified with the highest total phenolic content (TPC-57.70 ± 4.57 mg GAE/g), while sea parsley contained the lowest total phenolic content (13.44 ± 0.39 mg GAE/g). Moreover, bush mint was also identified with the highest antioxidant potential compared to other herbs. Thirty-seven phenolic metabolites were semi-quantified, including rosmarinic acid, chlorogenic acid, sagerinic acid, quinic acid, and caffeic acid, which were abundant in these selected plants. The most abundant compounds' pharmacokinetics properties were also predicted. This study will develop further research to identify these plants' nutraceutical and phytopharmaceutical potential.

Using Recombinant Superoxide Dismutase to Control Oxidative Stress in the Gastrointestinal Tract of Cyclic Heat-Stressed Pigs.

Climate change is associated with an increased frequency and intensity of heat waves, posing a threat of heat stress to pig production. Heat stress compromises the efficiency of pig production partly due to causing oxidative stress, intestinal dysfunction, and inflammatory responses. Superoxide dismutase is an antioxidant enzyme reported to reduce oxidative stress and inflammation. Therefore, this experiment aimed to investigate whether recombinant superoxide dismutase (rSOD) could ameliorate oxidative stress and inflammatory responses in heat-stressed grower pigs. Sixty-four female pigs (Large White × Landrace, 27.8 ± 1.65 kg, mean ± SD) were randomly allocated to a control diet (standard grower feed, CON) or the control diet supplemented with 50 IU recombinant superoxide dismutase (rSOD) for 14 days. After acclimation to the diet, pigs were then housed under thermoneutral (TN, 20 °C, 35-50% relative humidity) or cyclic heat stress conditions (CHS, at 35 °C: 9 a.m. to 5 p.m. and 28 °C: 5 p.m. to 9 a.m., 35-50% relative humidity) for 3 days. Heat stress increased respiration rate (RR), skin and rectal temperature (RR and RT) (p < 0.001 for all), and reduced plasma thyroid hormone concentration (p < 0.001). The amount of oxidized glutathione (GSH:GSSG) was increased in the jejunum and ileum of CHS pigs. In the jejunum, rSOD also increased the amount of oxidized glutathione in both TN and CHS pigs, without any change in endogenous SOD activity. In the ileum, rSOD prevented increases in oxidized glutathione formation in the CHS pigs only. Taken together, this may reflect increased oxidative stress in both the jejunum and ileum in CHS pigs. Alternatively, rSOD increased the conversion of reduced to oxidized glutathione independently of CHS, possibly reflecting an increased overall SOD activity due to the addition of exogenous SOD. In conclusion, the use of in-feed SOD enzymes at a dose of 50 IU/kg may be a useful strategy for preventing oxidative stress in pigs.

Identification and characterization of anthocyanins and non-anthocyanin phenolics from Australian native fruits and their antioxidant, antidiabetic, and anti-Alzheimer potential.

Polyphenols are vital bioactive constituents that have beneficial effects on human health. The aim of this study was to characterize the biologically active phenolic metabolites in Australian native commercial fruits (Kakadu plum, Davidson's plum, quandong peach, and muntries) and their antioxidant, α-glucosidase, and acetylcholinesterase inhibition activities. Polyphenols were measured through total phenolic content (TPC), total flavonoid content (TFC), total condensed tannin (TCT), and total monomeric anthocyanin content (TMAC). Moreover, different in-vitro biological assays (DPPH, ABTS, FICA, OH-RSA, α-glucosidase, and acetylcholinesterase inhibition activities) were conducted to measure the antioxidant, anti-diabetic, and anti-Alzheimer's potential of these selected fruits. LC-ESI-QTOF-MS/MS was implied for identification and quantification purposes. In this study, a total of 307 bioactive metabolites (51 phenolic acids, 194 flavonoids, 15 tannins, 23 other polyphenols, 5 stilbenes, 12 lignans, and 7 terpenoids) were putatively identified. A total of 41 phenolic compounds were quantified/semi-quantified. Kakadu plum was identified with a higher concentration of polyphenols and biological activities compared to Davidson plum, quandong peach, and muntries. Molecular docking was also conducted to discover the actual role of the most abundant phenolic metabolites in the α-glucosidase and acetylcholinesterase inhibition activities.