The aim of this preliminary study was to identify potential colour components, volatile and sensory attributes that could discriminate Pinot noir wines from five Australian winegrowing regions . The sensory analysis consisted of the Pivot© Profile method that was performed by wine professionals. A headspace solid-phase microextraction-gas chromatography-mass spectrometry method was used to quantify multiple volatile compounds, while the Modified Somers method was used for colour characterisation. Analysis of data suggested ethyl decanoate, ethyl 2-methylpropanoate, ethyl 2-methylbutanoate, in addition to decanoic acid as important contributors to the discrimination between regions. Similarly, wine hue, chemical age indices, total anthocyanin, and (%) non-bleachable pigment also discriminated wines between regions. The sensory analysis showed that wines from Mornington Peninsula were associated with the ‘red fruits’ aroma, ‘acidic’, and ‘astringency’ palate descriptors, while those from Adelaide Hills were associated with the ‘brown’ colour attribute. This study indicates regionality is a strong driver of aroma typicity of wine. Regionality (broadly referred to as terroir) is an important concept for winemakers and wine consumers, as it refers to how a wine is recognised on the basis of its geographical origin . This isWhen dealing with the regional expression of Pinot noir wine, gas chromatography-mass spectrometry (GC-MS) and spectral analysis can be used in order to discriminate samples obtained from different geographical locations ,3,4,5. FAs with aroma compounds and chromatic or colour structure components, relationships were hypothesised between the geographical origin of the grapes and the sensory features of the resulting wines. A descriptive analysis (DA) of 28 Pinot noir wines from California showed that samples from the cool Carneros appellation were more intense in the ‘fresh berry’, ‘berry jam’, ‘cherry’, and ‘spicy’ descriptors than those from the warmer Napa and Sonoma Valleys . DescripDescriptive analysis is a very robust and reliable sensory approach. However, it can be time consuming and costly, as panellists require extensive screening and training for up to several months . A more Vitis vinifera L.) is a very popular cultivar, particularly in cool-climate wine growing regions, where it produces fine, elegant, and expressive wines [Pinot noir . The wines were selected after consultation with noted Australian winemakers and wine judges . The priThe alcohol levels were commercially available and, thus, are expected to be ±0.5%, as per regulations. Titratable acidity (TA) and pH were determined by sodium hydroxide titration to an end point of pH 8.20 with a Fully Automated 59 Place Titrando System .The wines were sourced from 15 different vineyard sites, in general, on own-roots, drip irrigated, and Vertical Shoot Positioned .https://www.longpaddock.qld.gov.au/silo/).Australian geography is very heterogenous. This results in a range of climatic zones with many cool climate regions that are influenced by the Pacific Ocean and others influenced by the Tasman Sea. The main climate parameters for the growing season October 2017–April 2018, together with the altitude, for each of the five Australian regions examined in this study are summarised in n = 16 in total). Data were collected on personal digital tablets using the Compusense sensory software . Panellists generated appearance, aroma, and palate attributes to be compared against the pivot, as described in Thuillier et al. [A panel of 11 wine professionals  who regularly participate in wine shows or tasting events were convened (in January 2020) to evaluate the samples while using the Pivot© profile sensory method . The panr et al. . Social −1 (held for 14 min.) and then followed by series of temperature ramps. First ramp to 80 °C at 10 °C min.−1, second ramp to 160 °C at 20 °C min.−1, and third ramp to 260 °C at 10 °C min.−1 and held for 2 min. for a total run time of 45.5 min. The SPME vials and its contents were heated to 40 °C for 5 min. with agitation. The SPME fibre was exposed to the headspace in the sample for 15 min. and then desorbed in the injector (splitless mode) for 15 min. The injector was set at 260 °C. The MS quadrupole was set at 150 °C, the source was set at 230 °C, and the transfer line was held at 260 °C. Positive ion electron impact spectra at 70 eV were recorded in SIM and SCAN mode with a solvent delay of 4 min.A total of 28 fermentative compounds were analysed (in January 2020) by solid phase micro extraction (SPME)-GC-MS according to previously published method using Agilent Technologies Ltd. equipment  . One mL The raw data from Agilent’s ChemStation software (ver. E.02.02.1431) were processed by the MassHunter Workstation Software for Quantitative Analysis for GC-MS (ver. B.09.00). A stable isotope dilution analysis (SIDA) was used to determine the concentration of analytes in the samples. All of the target and qualifier ions of internal standards and analytes, in addition to their expected retention time, are reported in 13-norisoprenods α-ionone, β-ionone, and β-damascenone were analysed (in January 2020), as follows. Ten mL wine sample was transferred into 20 mL SPME vials with 2 g of NaCl and 50 µL of a combined d4-β-damascenone, d3-α-ionone, and d3-β-ionone internal standard solution. The GC-MS analysis was performed on an Agilent 7890 GC that was equipped with a Gerstel MPS2 autosampler and coupled to an Agilent 5977B N mass selective detector. The GC was fitted with an Agilent DB-5 MS  and ultra-high purity He was the carrier gas. The oven was started at 40 °C, held at this temperature for 1 min. then increased to 190 °C at 8 °C min.−1 and held at this temperature for 5.25 min. The vials and its contents were heated to 60 °C for 10 min. in the heater/agitator. The SPME fibre was exposed to the sample during this heating time and injected into a split/splitless inlet in splitless mode. The analytes were desorbed into a Supelco 0.75 mm ID sleeveless SPME liner  for 10 min., which was held at 200 °C. The purge flow to the split vent was 50 mL min−1 at 2.1 min. with the septum purge flow turned off. The mass spectrometer quadrupole temperature was set at 150 °C, the source was set at 230 °C, and the transfer line was held at 250 °C. EMV Mode was set to Gain Factor = 1.00 and th spectra were recorded in SIM mode. The ions monitored in SIM mode were: m/z 73, 179 and 194 for d4-β-damascenone and m/z 69, 175, and 190 for β-damascenone; m/z 112, 139, and 195 for d3-α-ionone and m/z 109, 136, and 192 for α-ionone; m/z 180, 181, and 195 for d3-β-ionone, and m/z 177, 178, and 192 for β-ionone.The Cw/v tartaric acid in 12% v/v ethanol adjusted to pH 3.4 with 5 M NaOH); (2) 1:10 dilution of wine in buffer 1 plus 0.375% w/v sodium metabisulphite; (3) 1:10 dilution of wine in buffer 1 plus 0.1% v/v acetaldehyde; (4) 1:50 dilution of wine in 1 M HCl. The samples were mixed and incubated at room temperature (20 °C) in the dark for at least 1 h and then read with a GENESYS 10S UV-Vis single cell Spectrophotometer  using the following wavelengths: 250, 270, 280, 290, 315, 420 and 520 nm. All of the parameters were determined from spectral data according to the calculations that were outlined in Mercurio et al. [The Pinot noir wine samples were analysed (in January 2020) according to the Modified Somers Assay, as previously described in Mercurio et al. . The wino et al. .p = 0.05 was used to test the differences between regional means. Principal component analysis (PCA) was performed with JMP . Separate PCA was performed for volatile and colour data sets. PCAs were developed using only the significant parameters (p < 0.05) to avoid fitting noise to the models.All of the data sets were analysed by one-way Analysis of Variance (ANOVA) using R . Equal variances of regional groups were verified by using Levene’s Test. Duncan’s multiple range-test at Raw Pivot© Profile results were downloaded into a spreadsheet and organized by ‘more than’ and ‘less than’ terms. Some lemmatisation was required during this analysis in order to reconcile terms that have similar meanings, for example ‘tannin’, ‘tannic’, ‘blocky tannin’, ‘tannins’, ‘soft tannin’, ‘hard tannin’ were all grouped under the one term: ‘tannin’. A data matrix was then created, with frequencies of the ‘less than’ terms being subtracted from the ‘more than’ terms. As this leaves some values negative (some attributes would be used as ‘less than’ more than ‘more than’), the results were adjusted to contain only positive values, with the most negative value added to complete set, leaving the most negative attribute zero and the remaining attribute’s positive. The results were then analysed by correspondence analysis (CA) using XLStat  to give a biplot of the samples and sensory attributes. Analysis was performed on appearance, aroma, and palate attributes individually, and then another with terms deemed to be relevant in separating the wines. This was determined by standard deviation of the attributes in the frequency table. Attributes with a standard deviation > 1.5 were included in the final CA biplot. In this instance, setting the minimum standard deviation at 1.5 excluded many of the fringe attributes and minimises noise that would have otherwise cluttered the analysis. For this analysis, the original appearance, aroma, and palate data were normalised to have the most negative score from all three modalities equal zero.A total of 31 analytes were identified and quantified in the headspace of 15 commercial, single-vineyard Australian Pinot noir wines. −1, the most abundant volatile compounds were the higher alcohols, such as 3-methylbutanol, 2-methylbutanol, and 2-methylpropanol. Higher alcohols are important contributors to the complexity of wine aroma. However, they give wines ‘solvent’ and ‘fusel’ characters at concentrations greater than 400 mg L−1, which may mask more elegant aromas that are associated with ethyl esters [Apart from acetic acid, which was found in the range of 600 to 700 mg Ll esters . On the Ethyl esters of fatty acids such as ethyl decanoate are yeast’ metabolism by-products, and their concentrations in wines largely depend on winemaking conditions  . By contN-(3-methylbutyl)acetamide, and N-(2-phenylethyl)acetamide discriminated the vineyard sites independent of vintage [There is little information in the literature with regards to the discrimination of Pinot noir wine on the basis of grape geographical origin and volatile compounds . Where t vintage . These a vintage .−1 and OAVs of 8–17. OAVs > 1 should theoretically have an impact on the aroma, although compounds with 1 > OAV > 0.5 have been shown to add wines complexity [As reported in mplexity . It coul−1, which is well below its odour threshold of 1000 µg L−1  [Decanoic acid was the only fatty acid to significantly change between regions . Specifi pH 3.2) . Despite pH 3.2) .p < 0.05) between regions.We quantified a number of chromatic and colour structure components in order to discriminate Australian Pinot noir wines on the basis of their geographical origin. The results for the 15 wine samples are summarised in −1 malvidin 3-O-glucoside) was significantly different between regions, with the wines from Southern Tasmania and Yarra Valley having higher levels than those from Mornington Peninsula  were higher in the wines from Adelaide Hills and Mornington Peninsula when compared to those from Yarra Valley, Southern and Northern Tasmania   non-bleachable pigment  . Hue (orasmania)  and therNon-supervised pattern recognition statistical analysis was employed through principal component analysis (PCA). The approach taken was not to generate a predictive model that could be applied to unknown samples, due to the experimental design being limited to three wines per five regions only. Rather, the aim was to identify possible patterns that were related to the classification factors.A total of 53 descriptors were generated by a panel of wine professionals : 10 appearance, 22 aroma, and 21 palate. The generated descriptors were analysed individually. Subsequently, the attributes that had a standard deviation < 1.5 in the frequency matrix  were incCombining all of the most important attributes from the appearance, aroma and palate biplot provides an overall characterisation of the Pinot noir samples . F1 and The wines from Northern and Southern Tasmania are spread out across the biplot exhibiting sensory characters that cover the broad spectrum of sensory attributes used to describe these Pinot noirs. For example, ST1, NT1, and YV1 seem, to some extent, to agree well with the ‘lower-right’ variables . Likewise, NT3 and MP3 agree well with the ‘astringent’ descriptor in the ‘upper-right’ quadrant. This is possibly due to the bigger geographical area of the Northern and Southern Tasmania regions as compared to the other regions, responsible for diluting a true regional effect.This preliminary study shows there are some regional chemical and sensory distinctiveness when it comes to Australia’s Pinot noir producing regions. From the volatile analysis, ethyl decanoate, ethyl 2-methylpropanoate, ethyl 2-methylbutanoate, and decanoic acid can be proposed as potential markers, suggesting that fermentation plays a significant role in the regional typicity of Australian Pinot noir wine. Some similarities were also apparent in terms of chromatic components and colour structure, in particular for the Adelaide Hills samples, which appeared to age more quickly than other regions.Some sensory similarities among wines from the same region were observed despite wines likely being produced using different winemaking techniques. In particular, wines from the Mornington Peninsula (and to a lesser extent those from Northern Tasmania) were associated with the ‘red fruits’ aroma, ‘acidic’, and ‘astringent’ palate descriptors, while the majority of wines from Adelaide Hills, Southern Tasmania, and Yarra Valley was associated with the ‘brown’ colour, ‘complex’ and ‘soft’ palate, ‘floral’ and ‘oaky’ aromas. This information, although preliminary, is important for those wine characters promoting regional typicity. Further analysis of a larger number of commercial samples and, more importantly, wines produced under standardised winemaking protocols is warranted to better understand this relationship.


Cannabis sativa L. (hemp). Leaves at three vertical positions in hemp crowns were examined from initial full leaf expansion until 42 days later. Photosynthetic capacity decreased as leaves aged regardless of crown position, light intensity, or photoperiod. Although leaves remained green, the photosynthetic capacity declined logarithmically to values of 50% and 25% of the maximum 9 and 25 days later, respectively. Plants grown under +450 μmol m−2 s−1 supplemental photosynthetically active radiation or enriched diffuse light responded similarly; there was no evidence that photoperiod or enriched diffuse light modified the gas exchange pattern. At approximately 14 days after full leaf expansion, leaf light levels >500 μmol m−2 s−1 decreased photosynthesis, which resulted in ≥10% lower maximum electron transport rate at ≥ 20 days of growth period. Furthermore, leaves were saturated at lower light levels as leaf age progressed (≤500 μmol m−2 s−1). Incorporating leaf age corrections of photosynthetic physiology is needed when estimating hemp primary production.We examined the aging of leaves prior to abscission and the consequences for estimating whole-crown primary production in  Three-dimensional crown growth and development, however, changes the within-canopy architecture and light environment as new leaves develop and self-shade one another  (An/Ci) analysis on leaves under two disparate light intensities (500 versus 1500 µmol photons m−2 s−1), representing light intensities separated by approximately 50% of the sun’s intensity at the earth’s surface. Although both light intensities were either close to (500 µmol photons m−2 s−1) or in excess (1500 µmol photons m−2 s−1) of hemp’s Ls  showed a similar Amax response as those among crown positions and treatments  resulted in a similar leaf-age-related response as compared to the logarithmic rate of decrease that was observed across crown positions . Whe. Whe2y-cmax and Jmax in hemp during the vegetative stage, supporting the need for age-related downscaling of these parameters. Overall, our results indicate that leaf age is an important temporal factor in hemp crowns. Our logarithmic modeling approach significantly simplifies the description of crown photosynthetic parameters as well as approximates the photosynthetic capacity of hemp leaves at various stages of ontogeny. From a canopy management view, our correlation models of leaf-age-related physiological deterioration simplify the description of age effects on hemp photosynthetic activity by permitting temporal scalars to modify photosynthetic parameters when predicting hemp crown productivity. This makes it possible to extract canopy gas exchange estimates that would otherwise be expensive and labor intensive to measure in the field. The next step is to incorporate leaf model parameterizations into a canopy-scale gas exchange model to effectively extrapolate the leaf-scale results to a whole canopy. In conclusion, we measured a progressive decline in VCannabis sativa ‘cherry wine’ were transplanted into 11 L bato buckets filled with horticulture perlite. Treatments consisted of two independent 1.5 × 2.5 m plots of 15 equally spaced plants: one under natural light and the other under natural light plus an extended photoperiod with supplemental light (+450 μmol m−2 s−1 photosynthetic photon flux (PPF) via high-pressure sodium lamps ). To ensure adequate nutrients and water, Peters’ Excel Cal-Mag (15-5-15) was applied twice daily at a strength of 200 ppm N through microemitter irrigation (360° substrate surface drip rings). All plants were allowed to acclimate for 14 days prior to crown morphology sampling.A preliminary study was conducted solely to understand hemp crown development and morphology at the Colorado State University Horticulture Center in Fort Collins, CO, USA. Thirty seed-based, two-week-old plugs of x, y, and z directions, in meters) on all plants six times over the course of the growing cycle to assess isotropic growth characteristics. Immediately after each of the six morphology measurement intervals, leaf area was estimated by a destructive harvest on a randomly selected plant per treatment. The crowns were equally subdivided into three vertical sections and all leaves per sector were removed and measured for total area . Fresh and dry weights were determined separately for leaves and stems, and dry mass was determined after being oven-dried at 70 °C for approximately 3–5 days.We sampled total plant height (HT), stem diameter (SD), stem length, and three-dimensional crown size  or extended photoperiod with supplemental light (n = 15) treatment. The extended photoperiod treatment maintained day length at 18 h; photoperiod varied in the natural treatment as day length changed. An additional 30 plants were transferred into an enriched diffuse light environment where reflective white plastic covered the floor surface and crown–crown competition among neighboring plants was minimal . The diffuse light enrichment was approximately 150% at the lowermost crown periphery (height above ground level ~0.4 m)  and the photoperiod varied as in the natural treatment.After the preliminary study, which allowed us to gain an understanding of the chronosequence of leaf development throughout a hemp crown, we conducted leaf gas exchange measurements on new and independent plant material from 7 August 2019 to 1 October 2019 at the Colorado State University Horticulture Center. Sixty feminized seeds of To examine the age-related photosynthetic capacity characteristics, three replicate plants per treatment located in the center of the plot were sampled for changes in leaf gas exchange traits. The south-facing second node branchlet leaf of the fifth main stem node from the substrate surface was tagged and repeatedly measured. Initially, the crown position encompassed the topmost fully expanded sunlit leaves and leaf age was defined as the days immediately after full leaf expansion. Due to isometric indeterminate growth during the vegetative phase of the growth cycle, the 10th and 15th node branchlets were added once the crown developed fully expanded leaves at the respective crown positions; that is, the crown was ultimately divided into three strata . Due to successive leaf production in hemp, this sampling regime facilitated the sequential addition of the middle and bottom crown positions at approximately 3 and 6 weeks into the study, together with repeated measurements of the same individual leaves. Periodically, measurements of leaves with contrasting ages and positions within a branch took place on a given sampling day. Gas exchange was repeatedly measured at each canopy position on fully expanded leaves of three replicate plants per treatment using a portable gas exchange system  fitted with a light- and environmentally controlled cuvette (Model PLC (U), PP Systems). The measurements were performed at a controlled leaf temperature of 25 °C and vapor pressure deficit (VPD) of 1.2 kPa.n) versus [CO2] (An/Ci) and photosynthetically active radiation (Qp) (An/Qp) were repeatedly run on three plants per treatment at three canopy positions. For the An/Ci curves, leaves were acclimated in the chamber for 5–10 min until An was stable at a controlled CO2 concentration (415 μmol mol−1) and light level (1500 μmol m−2 s−1). The CO2 response curve was constructed at CO2 levels of 415, 300, 200, 150, 100, 50, 25, 450, 500, 800, 1000, 1500, and 415 μmol CO2 mol−1. After the completion of the An/Ci curve, CO2 concentration was kept constant at 415 μmol mol−1 and Qp was sequentially lowered from 1500 to 1200, 1000, 800, 600, 400, 200, 175, 150, 125, 100, 80, 60, 40, 25, 20, and 0 μmol m−2 s−1. Due to the time consumption of generating a An/Qp and An/Ci curve per leaf and the continuous addition of new leaves as canopy positions developed, the younger leaf sample size ended up being larger than that of older leaves. Immediately after gas exchange, five replicate light absorptance samples were recorded at that leaf with a SPAD meter and averaged . The SPAD values were used to correct ϕa to ϕ as described in [n/Ci response curve generated at 500 μmol m−2 s−1 Qp to cover the effects of excess light on photosynthetic function.Response curves of net photosynthesis  were placed parallel to the north/south row orientation adjacent to the plant stems. A third line sensor was placed 0.5 m above the canopy. The line sensors sampled PAR every minute and then recorded a 15 min average .cmax) and maximum rate of electron transport for RuBP regeneration (Jmax) under saturating light. The An/Ci data were analyzed as per the default fitting method “fitaci” function of the “plantecophys” package [c), RuBP regeneration (Wj), and regeneration of inorganic phosphate (Tp)—can limit CO2 assimilation. Tau (τ) represents the specificity factor for Rubisco [L), comprising mainly of processes other than photorespiration, refers to the release of CO2 in the light [2 compensation point (Γc) was estimated from the intersection of the regression line with the x-axis. Equation (1) was fitted separately for An/Ci curves at 1500 and 500 μmol m−2 s−1.The model of Farquhar et al.  was used package  accordin package :An =  [a underestimation from data in the nonlinear region (≥125 μmol m−2 s−1) [c) was estimated from the intersection of the regression line with the x-axis (An = 0), and Rd was measured at the end of the An/Qp curve. Quantum yield for CO2 (ϕCO2) was derived from ϕa by correcting ϕa for the percentage leaf absorptance of Qp as described in Bauerle et al. [n/Qp curves.Photosynthesis versus Qs et al.  using lem−2 s−1)  and prevm−2 s−1) . The lige et al. . Equatiomax, ϕCO2, Ls, Jmax, Vcmax, Γc, and TPU with leaf age can be expressed in the following leaf-age-based logarithmic scalar function :a and b are the fit constants of the logarithmic (ln) least-squares regressions between the rate of change for Amax, ϕCO2, Ls, Jmax, Vcmax, Γc, and TPU and leaf age (La), respectively. From a causal view, the form of Equation (3) introduces biological function by allowing each physiological parameter to achieve its theoretical maximum at the time of full leaf expansion. The model was validated for ϕCO2 via the known requirement of 8 mol of photons to reduce 1 mol of CO2 where the theoretical maximum ϕCO2 = 0.125. Due to cyclic photophosphorylation, 0.125 is then reduced to ~0.112 and further reduced to ~0.092 by means of CO2 released through photorespiration measured at atmospheric O2 levels [2 of ~0.092 would then decrease further due to a deterioration in physiological function [The functional form of the rate of change in A2 levels ,12,40. An/Qp and An/Ci curves was 27 leaves . Of the nine leaves per treatment, n = 3 replicate leaves were at each of three crown positions per treatment. Leaves ranged in age from 1 to 42 days. The curve parameters were evaluated at three-day age interval ranges. A one-way ANOVA with the least significant difference at a significance level of p > 0.05 was used to assess photosynthetic parameters among treatment factors per three-day interval and was found to not be significantly different among treatments and crown positions. Samples were pooled among treatments and crown positions and subsequently binned to build a general leaf age effect equation per photosynthetic parameter. Logarithmic least-squares and linear regression was fitted between photosynthetic parameters and leaf age, treating age as a continuous variable. In addition, we investigated exponential, polynomial, and power correlation analysis to assess the strength of the coefficient of determination (r2). For leaf ages beyond day 17, there was not a balanced design at every measurement interval due to the time consumption of generating an An/Qp and two An/Ci curves per leaf; so after day 17, we used a one-way ANOVA with replicate and treatment, nesting replicate within treatment.The gas exchange experiment was designed to allow us to discriminate between effects due to leaf age, effects due to crown position, and any subsequent interactions due to light environment . The total sample size for A


The circadian rhythm plays a chief role in the adaptation of all bodily processes to internal and environmental changes on the daily basis. Next to light/dark phases, feeding patterns constitute the most essential element entraining daily oscillations, and therefore, timely and appropriate restrictive diets have a great capacity to restore the circadian rhythm. One of the restrictive nutritional approaches, caloric restriction (CR) achieves stunning results in extending health span and life span via coordinated changes in multiple biological functions from the molecular, cellular, to the whole–body levels. The main molecular pathways affected by CR include mTOR, insulin signaling, AMPK, and sirtuins. Members of the family of nuclear receptors, the three peroxisome proliferator–activated receptors (PPARs), PPARα, PPARβ/δ, and PPARγ take part in the modulation of these pathways. In this non-systematic review, we describe the molecular interconnection between circadian rhythm, CR–associated pathways, and PPARs. Further, we identify a link between circadian rhythm and the outcomes of CR on the whole–body level including oxidative stress, inflammation, and aging. Since PPARs contribute to many changes triggered by CR, we discuss the potential involvement of PPARs in bridging CR and circadian rhythm. The circadian  clock is a system conserved from cyanobacteria to humans, which generates internal oscillations with a near 24 h periodicity coordinating bodily processes in an anticipatory fashion. In mammals, the circadian clock governs nearly all aspects of physiology and behavior, including hepatic metabolism, cardiovascular activity, gastrointestinal tract activity, blood pressure, endocrine secretions, body temperature, renal function, and sleep–wake rhythm ,2,3,4. Tad libitum feeding conditions are driven by the daily pattern in food intake. Nearly 10% of the hepatic transcriptome oscillations are under the control of the cell–autonomous circadian clock and feeding schedule. Out of nearly 3000 of hepatic transcripts oscillating daily, fasting suppresses the oscillation of >80% of them , oxidation of GSH by reduction of H2O2 [GSH peroxidase (GPx)], and conjugation of GSH [GSH S–transferase (GST)] to various electrophilic substrates [+ and its redox partner NADPH, which were mentioned previously as they are inseparably connected with SIRTs. Moreover, in cultured rat fibroblasts high NAD+ and NADP+ levels decrease the binding capacity of CLOCK:BMAL1 heterodimers to E–box targets, whereas high levels of the reduced NADH and NADPH strongly enhances their binding in concert with BMAL1 acetylation, leading to increased expression of Pers, Crys, and other circadian genes [Reduced glutathione (GSH), in contrast to oxidized GSH (GSSG), is one of the most important non–enzymatic antioxidants. GSH acts as a scavenger of free radicals, supports the reduction of Hbstrates ,507,508.an genes . Additioan genes . Moreovean genes . TherefoPparα KO mice, increased oxidative stress occurs at an earlier age compared to WT animals [Sod–1, glutathione peroxidase 1 and 3 (GPx1/3), Cat, and uncoupling protein 2 (Ucp2) [In  animals  and dosi animals . WY–1464 animals . In a ge2 (Ucp2) .2− levels in cardiac muscle [2− accumulation in the mitochondria and aids in the export of ROS from mitochondrial to the cytosol [PPARγ also regulates the expression of several antioxidant and pro–oxidant enzymes, as well as oxidative stress–related proteins, including GPx3 , CAT 51,515, andc muscle . Furtherc muscle ,520,521.c muscle . Furtherc muscle , which p cytosol . Moreove cytosol ,525,526. cytosol ,528.Nuclear factor erythroid 2–related factor 2 (NRF2), a redox–sensitive transcriptional factor that serves as the master regulator of the oxidative stress response, cryoprotection against oxidative as well as electrophilic stress and takes part in inflammation suppression  containsIfnγ), granzyme B (GrB), perforin (Prf), and Tnfα oscillate showing the highest expression at the break of the dark and the light phase which coincides with the cytolytic activity of splenic natural killer cells [Salmonella enterica serovar Typhimurium there is a more efficient bacteria clearance for infection done 4 h after the beginning of the dark phase compared to 4h into the light phase. Moreover, levels of antimicrobial peptide lipocalin–2 in the intestine are higher during the day than during the night [The immune system as one of the key factors indispensable for survival has been intensively investigated in the context of circadian rhythms. The expression levels of toll–like receptors (TLRs) display significant circadian fluctuations in the mouse jejunum . In the er cells ,536,537.er cells . Accordihe night . Importahe night . In the he night .The central pacemaker can also regulate oscillations of inflammatory factors in peripheral organs as local splenic sympathectomy alters daily variations of cytokines, TNFα, and cytolytic factors, GRB, and PRF, in natural killer cells and splenocytes . A lesioα, IL1β, and Il–6 [α impacts Il–1β, and Il–6 [α regulates IL–6 [hi inflammatory monocytes [Clock KO mice exhibit reduced cytokine production in macrophages in response to the LPS challenge or Salmonella Typhimurium infection [Ror–α in mice leads to a stronger LPS–induced inflammation with higher levels of Il–1β, IL–6, and macrophage inflammatory protein–2 in bronchoalveolar lavage fluids from mice lungs [α and IL–6 upon activation [Per2 KO mice are more resistant to the LPS challenge compared to WT mice [Cry1 and Cry2 also leads to increased inflammation, a permanent elevation of proinflammatory cytokines in a cell–autonomous manner, and constitutive activation of NF–κB and PKA signaling [Sod and other anti–inflammatory genes [κB to blunt age–triggered increases in COX–2 levels and activity. CR suppresses the generation of iNOS, IL–β, IL–6, TNFα, and prostanoids, such as TXA 2, prostacyclin 2, and prostaglandin E2 [κB, signal transducer and activator of transcription 1 (STAT1), and activating protein–1 (AP–1) [Clock genes have been shown to modulate the expression of multiple inflammatory cytokines. CRY affects TNFand Il–6 , RORα imand Il–6 , PER2 moand Il–6 , and Il–and Il–6 , Rev–ERBtes IL–6 , MCP-1 ates IL–6 , while Bonocytes  and reduonocytes . Consequnfection . On the ce lungs . Moreovetivation . Per2 KO WT mice . KO or signaling . Importaignaling . NF–κB iry genes ,562,563.andin E2 ,485 whilandin E2 . PPARs i1 (AP–1) ,566,567 1 (AP–1) ,565. Mor1 (AP–1) ,569. Howα are mostly based on its interaction with NF–κB. Accordingly, PPARα KO results in premature oxidative stress and an aggravated inflammatory response as well as a stronger age–dependent increase in oxidative stress and NF–κB activity [α ligands induce NF–κB, whereas low amounts reduce NF–κB activation [C4bp, C9, Mbl1, Orm1, and Saa4) responsive to inflammatory cytokines [The anti–inflammatory properties of PPARactivity ,570. In activity ,571. Imptivation . PPARα cytokines .κB. The activation of PPARβ/δ protects skeletal muscle against metabolic disorders also by means of counteracting the diet–induced activation of NF–κB as well as expression of iNOS and intercellular–adhesion–molecule–1 (ICAM–1) [α secretion. Accordingly, overexpression of PPARβ/δ substantially inhibits TNFα expression, while the absence of PPARβ/δ enhances LPS–induced TNFα production in cultured cardiomyocytes [PPARβ/δ is also capable of regulating NF–(ICAM–1) . GW0742,myocytes . In the myocytes .Pparγ polymorphism is associated with changes in the onset of diseases with an inflammatory background including multiple sclerosis [PPARγ is one of the most potent anti–inflammatory factors with extensive research proving its properties. PPARγ agonists diminish inflammatory bowel disease symptoms and are instrumental in the therapy of ulcerative colitis and Crohn’s disease ,579,580.clerosis , T2D [42clerosis , vasculaclerosis , as wellclerosis ,587. PPAclerosis , Bcl2 [3clerosis , c–Myc, clerosis , Cox–2 [clerosis ,591, iNOclerosis , and Apcclerosis ,594. Mosclerosis ,591. Conclerosis ,596. TheClock and Bmal1 in the hamster SCN [Circadian clocks have been implicated in the control of aging and aging–associated pathologies via regulation of oxidative stress, cell cycle, cell death, and DNA damage response . Importaster SCN  and resuster SCN .Bmal1 KO mice have reduced lifespans and display various symptoms of premature aging including sarcopenia, cataracts, lesser subcutaneous fat, and organ shrinkage [Clock KO mice being reduced by 15% and 20%, respectively, compared to WT mice. This phenotype is accompanied by a high rate of development of age–related pathologies, cataracts, and dermatitis [Bmal1 KO mice. Based on wheel–running activity, the circadian rhythm in Per1, 2 KO mice is out of control. These animals also display signs of premature aging including loss of soft tissues and kyphosis, a more rapid decline in fertility and litter size in each successive breeding, as well as a significant increase in neoplastic and hyperplastic phenotypes [Per1 and Per2 were proposed as longevity–associated candidate genes based on (i) the KO phenotype in mice, (ii) comparison of the activity of a group of genes with differential expression in several tissues such as the liver, brain, skeletal muscles of mice subjected to CR and (iii) Per gene expression in the long–lived dwarf mice [tau, resulting in 24 h light–dark cycles, life span is shortened. However, in aged animals, longevity can be extended by fetal suprachiasmatic implants which restore higher amplitude rhythms [hrinkage . Deficiermatitis . Howeverenotypes . Per1 anarf mice ,611. In  rhythms ,613,614.Oscillation disturbance by weekly reversal of the light–dark cycle reduces the survival time of cardiomyopathic hamsters . TransplDrosophila enhances the amplitude of cycling of most circadian clock genes, including Tim, in peripheral tissues [Per and Tim KOs attenuate the CR–dependent changes in lifespan extension and fat metabolism [Bmal1 KO mice [In young mice, 2626 liver genes present a rhythmic expression, while in old mice this number is reduced to 1664 genes, including 1575 genes that are rhythmic in both groups and comprise the core clock genes. CR increases the number and amplitude of rhythmic genes in both young and old mouse livers . Besides tissues . Per andtabolism . Similar KO mice . TherefoAging is associated with decreased metabolic rate, impaired mitochondrial function, reduced insulin resistance, dysfunctional lipid metabolism, immunosenescence, increased oxidative stress, and lowered hormonal secretion ,621,622.α expression and constitutive activation of some of its target genes including genes involved in β– and ω–oxidation of FAs [α in the muscle and PPARβ/δ in the liver and skeletal muscle as WT CR mice [α target gene, extends lifespan in mice and blunts GH/IGF–1 signaling in the liver [Most of the multiple traits of aging are related to PPARs and mTOR activity, oxidative stress, inflammation, and metabolism . Furthern of FAs ,631. Gro CR mice . Fitting CR mice . Furtherhe liver . TherefoImportantly, not only feeding schedule or feeding restriction (CR) but also nutrients entrain the circadian rhythm. A balanced combination of carbohydrate and protein induces phase advance in the liver clock, whereas the application of a single macronutrient  had no significant impact . Similar


We present an unusual case of a previously healthy 74-year-old man who presented with diffuse weakness, severe myalgias, petechial palmar rash and hypotension, but without fever, altered mental status, nuchal rigidity or headache, who was ultimately found through PCR testing to have meningococcal meningitis. Despite advances in vaccination and treatment, bacterial meningitis remains a deadly disease . The eldWe present a case of a previously healthy 74-year-old man who presented with diffuse weakness, severe muscle aches, rash and hypotension, but without fever, altered mental status, neck rigidity or headache, and who was ultimately found to have meningococcal meningitis.Neisseria meningitidis in the Los Angeles area  (noted to be 229 000 2 weeks prior to presentation) and a leukocytosis of 17 020 μl–1 [normal range 416 000–995 000], 88 % neutrophils [normal range 40–60 %], concerning for bacterial infection. Additional laboratory studies showed an acute kidney injury with creatinine of 2.3 mg dl–1 (1.0 2 weeks prior), an elevated erythrocyte sedimentation rate of 46 mm h–1, creatinine kinase elevated at 197 U l–1, fibrinogen 561 mg dl–1, lactate 40 mg dl–1, HIV- 1/2 Ag/Ab negative, complement normal, and cortisol 54 mcg dl–1. C-reactive protein was found to be abnormal at 26.8 mg dl–1. His blood pressure responded to volume resuscitation with normal saline. The patient was started on vancomycin and piperacillin/tazobactam, and he was admitted to the intensive care unit. Blood cultures were then drawn and were sterile.On presentation he was found to be afebrile with a temperature of 36.7 °C. His heart rate was 67 beats per minute and blood pressure 95/61 mmHg, which dropped to 65/48 mmHg within an hour of presentation. He was fully oriented and interactive, and his exam was notable for 4/5 weakness in bilateral upper and lower extensor and flexor muscles, severe total body myalgias, and a petechial rash on the palms and soles of his feet. His neck was supple, and the remainder of his neurological exam was within normal limits. Laboratory studies were notable for a platelet count of 139 000 μl–1 glucose (serum glucose 152 mg dl–1) and 93 mg dl–1 protein. CSF Gram stain showed no bacteria with many white blood cells, and CSF culture was negative . On hospital day 6 the BioFire FilmArray PCR panel demonstrated N. meningitidis. The patient recovered well with no residual neurological deficits and was discharged home on hospital day to complete a 14-day course of ceftriaxone.Then, 36 h after initial presentation and administration of antibiotic, the patient had a fever of 38.1 °C. MRI brain and cervical spine showed ‘layering restricted diffusion in the lateral ventricles bilaterally…[concerning for] layering pus in the setting of ventriculitis.’ Piperacillin /tazobactam was switched to meropenem, and voriconazole and ampicillin were added to his antimicrobial regimen. Attempts at beside lumbar puncture failed secondary to degenerative spinal disc disease, and lumbar puncture was finally performed on hospital day 4 under CT guidance. Cerebral spinal fluid (CSF) showed 3000 cells per cubic millimeter (cmm) red blood cells, 1654 cmm white blood cells , 81 mg dlGiven the high and swift mortality associated with bacterial meningitis, learning from unusual presentations may aid faster diagnosis in the future. This case illustrates several important clinical points, including an unusual presentation of meningitis in an elderly individual, the utility of PCR-based testing of CSF especially with prior antibiotic use, and the importance of meningococcal vaccination.Although the sensitivity of the classic triad of fever, neck stiffness and altered mental status is less than 50 %, when headache is added as a diagnostic criterion the sensitivity of 2/4 symptoms rises to 99–100 % . Of intein vitro PCR assay that simultaneously tests for 14 common pathogens in meningitis and encephalitis [Additionally, the identification of a specific pathogen hinged upon the BioFire FilmArray PCR panel. The FA Meningitis/Encephalitis Panel is a qualitative phalitis .In our case, likely because of delay in lumbar puncture, Gram stain and CSF culture were unrevealing. While CSF cell count and chemistry led to the diagnosis of bacterial meningitis, the identification of a specific pathogen using PCR not only led to targeted therapy but also directed important post-exposure prophylaxis (PEP) of close contacts.N. meningitidis. One study showed that one third of CSF samples with meningococcal infection were sterile within 1 h of antibiotic administration, and all samples were sterile within 2 h [While CSF culture remains the diagnostic gold standard for bacterial meningitis , the timthin 2 h . Even whthin 2 h .N. meningitidis compared to other common pathogens, ranging from 30–89 % [As an additional diagnostic tool, CSF Gram stain can rapidly and accurately identify a causative bacterium in 60–90 % of patients with community-acquired bacterial meningitis, with a specificity of >97 % . However 30–89 % .PCR technology has become cheaper and faster, making it a more accessible clinical tool. PCR tests, through amplification of a specific genetic target of a micro-organism, is generally more sensitive than conventional culture-based methods and is not affected by the viability of the micro-organisms, especially those that are fastidious and hard to grow in the lab. One study used a latent class analysis model, and found the sensitivity and specificity estimates were: culture, 81.3 and 99.7 %; Gram stain, 98.2 and 98.7 %; and real-time PCR, 95.7 and 94.3 %, respectively. The sensitivity of Gram stain and PCR did not change significantly when antibiotics were also present in the CSF . The widN. meningitidis among young men who have sex with men (MSM). Although our patient was MSM, he endorsed a long-term, monogamous marriage, and denied what are considered typical risk factors such as bars or dating apps, and he had not received any meningococcal vaccinations. In 2016, Los Angeles County reported 17 cases of invasive meningococcal disease (up from 12 in 2015); 9 (60 %) in MSM, with a median age of 32 (range 16–76) [N. meningitidis among MSM [In addition to increasing awareness of unusual presentation of meningococcal meningitis in the elderly, this case also illustrates the benefit of expanded vaccination. This patient presented in Los Angeles during a known outbreak of  up from  in 2015; up from  in 2015;e 16–76) . It is umong MSM . Based omong MSM .N. meningitidis highlights the utility of PCR-based testing of CSF, and advancements in reliable, rapid detection.This unusual presentation of


Tree nuts play an important role in healthy diets, but their economic value and nutritional quality may be affected by their size and paternity. We assessed relationships between nut size and kernel recovery, the incidence of whole kernels, fatty acid composition and mineral nutrient concentrations in three macadamia cultivars, “Daddow”, “816” and “A4”. We determined to what extent differences in nut size and quality were the result of different levels of cross- or self-paternity. Small nuts of all cultivars had lower kernel recovery than large nuts, and small nuts provided lower incidence of whole kernels in “Daddow” and “A4”. Small kernels had a lower relative abundance of the saturated fatty acid, palmitic acid, in all cultivars and higher relative abundance of the unsaturated fatty acid, oleic acid, in “Daddow” and “A4”. Small kernels had higher concentrations of many essential nutrients such as nitrogen and calcium, although potassium concentrations were lower in small kernels. Most nuts arose from cross-pollination. Therefore, nut size and kernel quality were not related to different levels of cross- and self-paternity. Identified cross-paternity was 88%, 78% and 90%, and identified self-paternity was 3%, 2% and 0%, for “Daddow”, “816” and “A4”, respectively. Small macadamia kernels are at least as nutritious as large macadamia kernels. High levels of cross-paternity confirmed that many macadamia cultivars are predominantly outcrossing. Macadamia growers may need to closely inter-plant cultivars and manage beehives to maximise cross-pollination. Tree nuts play an important role in healthy human diets, providing beneficial fatty acids, proteins and essential mineral nutrients ,2,3. MosThe fatty acid composition and nutrient concentrations of tree-nut kernels can vary greatly among cultivars ,26,27,28Macadamia integrifolia, M. tetraphylla and hybrids). Macadamia flowers are bee-pollinated and partially self-incompatible [Relationships between kernel size, nutritional composition, and cross- or self-paternity are poorly understood for some nuts, including the subtropical tree nut, macadamia , palmitic acid, than kernels from large nuts . These rOur results showed that small macadamia kernels have a fatty acid profile that is at least as healthy as large macadamia kernels. However, the differences in fatty acid profile between small and large kernels were much less than the differences between macadamia cultivars ,72,73 anKernels from small nuts had higher concentrations of many mineral nutrients than those from large nuts . NutrienPotassium concentrations, in contrast to the other mineral nutrients, were lower (by 8%–12%) in kernels from small nuts . These rBoth small and large macadamia nuts arose mostly from cross-pollination . The oveThe high level of cross-paternity in “Daddow” was particularly interesting because “Daddow” nuts were sampled in the middle of a 48-row single-cultivar block. Here, at least 88% of nuts were fathered by trees that were at least 200 m away. These results extend previous findings that 85%–90% of nuts in the middle of a 27-row block of “A16” trees arose from cross-pollination . In thatThe finding that many macadamia cultivars are predominantly outcrossing has enormous implications for the management and design of macadamia orchards. Few macadamia growers rank pollination as a major factor that limits production, with most Australian growers nominating more-clearly observable factors such as storm and hail damage, pest outbreaks, and hot or dry weather . Macadam°47′53″ S 152°17′36″ E) had a red-dermosol soil, while Site 2 (24°56′6″ S 152°21′16″ E) had either a yellow/brown kandosol or a red kandosol soil depending on the location within the orchard [We sampled nuts from two commercial macadamia orchards near Bundaberg, Queensland, Australia. Bundaberg has a humid subtropical climate with wet summers and dry winters, mean annual rainfall of 1009.5 mm, and mean minimum–maximum temperatures of 21.5–30.4 °C in January and 10.3–22.3 °C in July . Site 1  orchard . Trees aWe investigated three cultivars, with each cultivar sampled at one site. Cultivar “Daddow” was sampled at Site 1. Cultivars “816” and “A4” were sampled at Site 2. “Daddow” trees are typically large and rounded, with an open canopy that progresses to a dense canopy with age, and with medium-sized nuts . “816” tThe middle row of each cultivar was selected to increase the likelihood of finding self-pollinated nuts. We sampled six trees from each cultivar, commencing with the fifth tree from the end of each row at Site 1 and the twentieth tree from the end of each row at Site 2, and then sampling every tenth tree. We collected samples from the orchard floor under each tree on two occasions during the harvest season of each orchard. “Daddow” nuts were collected at Site 1 on 26 Mar 2018 and 18 Apr 2018. Cultivar “816” and “A4” nuts were collected at Site 2 on 27 Mar 2018 and 10 May 2018.We dehusked the fruit from each sampled tree before drying the nuts at 37 °C for 2 d, 45 °C for 2 d and 57 °C for 2 d . Non-comWe recorded nut-in-shell (NIS) mass for each nut, before cracking it using a manual nutcracker . Kernel mass, and whether the kernel remained whole or split into halves, was also recorded. Kernel recovery was calculated as the percentage of NIS that was comprised of kernel. Each individual kernel was then crushed and dissected into three representative sub-samples of at least: (a) 500 mg for fatty acid analysis; (b) 300 mg for mineral nutrient analysis; and (c) 30 mg for paternity analysis.We analysed fatty acids as their methyl esters from 154 “Daddow”, 146 “816”, and 160 “A4” kernels. The first 144 kernels of each cultivar were selected randomly, and additional kernels were selected based on known paternity see , below. v/v) and 25 µL of HCl  to 1 µL of the extracted oil. The mixture was incubated for at least 20 h at 65 °C before adding 0.5 mL of hexane and 0.5 mL of deionised water and shaking for 30 sec. The layer of deionised water was removed manually before adding another 0.5 mL of deionised water to the hexane. The layer of deionised water was removed, and anhydrous Na2SO4 was added to remove any remaining water from the hexane. The hexane was removed, avoiding disruption of the Na2SO4 residue, and fatty acids were analysed by gas chromatography–mass spectrometry   [We added 0.7 mL of anhydrous methanol dibutyl hydroxytoluene solution  by combustion analysis using a LECO CNS 2000  ,110. We We collected a leaf from each sampled tree, from which an approximately 50 mg subsample was used to confirm cultivar identity. We also identified potential pollen–parent cultivars within 1.5 km of the study sites, and we collected leaf samples from each of these cultivars. Seventeen cultivars were identified as candidate pollen parents and incorporated into a reference library for paternity assignment. This reference library contained no additional cultivars to those established in the orchard at Site 2; i.e., candidate pollen–parent cultivars were “A4”, “A16”, “A29”, “A38”, “A203”, “A268”, “Daddow”, “Own Venture”, “246”, “344”, “660”, “741”, “814”, “816”, “835”, “842” and “849”.2, 0.2 μM of each oligonucleotide primer and 6.7 μL sterile water. PCR was performed with initial denaturation at 94 °C for 2 min, followed by 35 cycles of 94 °C for 10 s, 55 °C for 10 s and 70 °C for 1 min, then final elongation at 70 °C for 5 min. Samples were maintained at 10 °C.We froze each leaf or kernel subsample in liquid nitrogen and ground it to a powder using an MM2000 TissueLyser  after adding disposable 0.1 and 2.3 mm diameter zirconia/silica beads  . DNA wasWe separated the PCR products by agarose gel electrophoresis and viewed them under ultra-violet light on a Gelscan visualiser . PCR products were diluted with molecular-grade water . The PCR dilution (1 μL) was added to a solution containing 9 μL Hi-Di and 0.1 μL GS-600 LIZ  and injected into an AB 3500 Genetic Analyser . We determined allele sizes relative to GS-600 LIZ using GeneMarker v.2.6.3 . Scoring was cross-checked manually to ensure accuracy and consistency . We idenWe analysed paternity of kernels by the logarithm of odds (LOD) method using Cervus 2.0 software . The LOD for each possible pollen parent was calculated by taking the natural log of the overall likelihood ratio. A positive LOD meant that the candidate cultivar was more likely to be the true pollen parent than a cultivar selected randomly from the reference library. Cultivars with a negative LOD were excluded as pollen parents for each kernel. We then assigned nuts as cross- or self-pollinated if all remaining pollen parents were either cross-pollen cultivars or the self-pollen cultivar, respectively. Remaining nuts were then assigned as cross- or self-pollinated if both cross- and self-cultivars were possible pollen parents, but one candidate pollen parent had strict-level confidence (95%) of the LOD. Kernels were considered “unassigned” if both cross- and self-pollination remained possible but the confidence value for the primary-candidate pollen parent was less than 95%. Kernels were considered “mismatched” if no parent pair from the reference library could explain its genotype.t-tests. The data sets had normal distributions and homogeneous variances, and so data transformation was unnecessary. Means and interactions were regarded as significantly different at p < 0.05. Means are reported with standard errors.We analysed data for nut-in-shell mass, kernel mass, kernel recovery, fatty-acid relative abundance, kernel nutrient concentrations and kernel nutrient contents by 3-way analyses of variance (nut size × harvest time × tree) using IBM SPSS Statistics v. 25. Nut size × harvest time and nut size × harvest time × tree interactions were generally not significant and so reported data were pooled across the two harvest times. Whole kernel and paternity data were analysed by paired Small macadamia nuts provided lower kernel recovery and whole-kernel incidence than did large nuts, which reduces their financial value. However, higher mineral nutrient concentrations and more nutritionally-beneficial fatty acid profiles ensured that kernels from small macadamia nuts were at least as healthy in human diets as kernels from large nuts. Relationships between nut size and kernel quality were not the result of differences in cross- and self-paternity levels between small and large nuts. Macadamia trees were predominantly outcrossing. Most nuts of all three cultivars, “Daddow”, “816” and “A4”, were cross-pollinated, even in the middle of a 48-row block of a single cultivar. Growers might need to consider inter-planting cultivars closely and managing bee hives to maximise the dispersal of pollen from one macadamia cultivar to another.


Saccharina cultivar ‘Haiyi No.1’ (Saccharina japonica × latissima). This mitogenome had a circular mapping organization with the length of 37,657 bp and contained 66 genes including 35 protein-encoding genes, 3 rRNAs, 25 tRNAs, and 3 open reading frames (ORFs). The AT content accounted for 64.7% of the entire mitogenome. The gene content and gene order were consistent with those reported Saccharina species and cultivars. Phylogenetic analysis showed the phylogenetic relationship among Chinese Saccharina cultivars and demonstrated that ‘Haiyi No.1’ had a closer relationship with Pingbancai which strongly supported its genetic source.Here, we reported the complete mitogenome of  Saccharina  species are the economically important brown seaweeds (Kain Saccharina cultivars been bred in China which has contributed significantly to the mariculture. ‘Haiyi No.1’ bred from the ‘Pingbancai’ which is an intraspecific hybrid of ‘Fujian’ (Saccharina japonica) and ‘901’ (Saccharina japonica×latissima). At present, more than 10 mitogenomes of Chinese Saccharina cultivars have been completed. However, genomic information of ‘Haiyi No.1’ is limited. Here, we characterized the complete mitogenome of ‘Haiyi No.1’ and constructed phylogenetic analysis to provide new molecular data for the molecular study.eds Kain . There a‘Haiyi No.1’ individuals were collected from Rongcheng, Shandong Province, China , and then specimen (sample accession number: 2016070089) was deposited in the Culture Collection of Seaweed at the Ocean University of China. Subsequently, the homologous PCR amplification and primer walking strategy were used , with a nucleotide composition of 28.4% A , 14.7%C (5536), 20.6% G (7757), and 36.3% T , and had an overall AT content of 64.7%. This mitogenome encoded 3 rRNA genes , 25 tRNA genes, 35 protein-encoding genes, and 3 ORFs . The protein-encoding regions were 28,667 bp in length accounting for 76.1% of the whole mitogenome. Among these 66 genes, only rpl2, rpl16, rps3, rps19, tatC, and orf130 were encoded on L-strand. The universal genetic codes were used and all start codons were ATG. Of the 35 protein-encoding genes and 3 ORFs, 26 (68.4%) terminated with TAA codon, higher than that for TAG  and TGA . Moreover, all tRNA genes were provided with standard clover-leaf secondary structures. Gene content and gene order showed a high level of conservation with those reported Saccharina mitogenomes  showed there were only three mutation sites which indicated high conservatism at the intergenic region.Saccharina and Laminaria algae was conducted to reconstruct the phylogeny (Ectocarpus siliculosus served as the out-group. Phylogenetic analyses showed that all algae were divided into two branches: Saccharina and Laminaria. All Chinese cultivars including ‘Haiyi No.1’ belonged to the Saccharina lineage. ‘Haiyi No.1’ first clustered with ‘Pingbancai’, ‘Zaohoucheneg’ and ‘Ailunwan’ supporting its genetic origin. The phylogenetic tree in this work also provided more information for the current understanding of genetic relationships among Chinese Saccharina cultivars.Bayesian analysis based on the whole mitogenomes of the 18 hylogeny . Ectocar


The proliferator-activated receptor γ (PPARγ), a member of the nuclear receptor superfamily, is one of the most extensively studied ligand-inducible transcription factors. Since its identification in the early 1990s, PPARγ is best known for its critical role in adipocyte differentiation, maintenance, and function. Emerging evidence indicates that PPARγ is also important for the maturation and function of various immune system-related cell types, such as monocytes/macrophages, dendritic cells, and lymphocytes. Furthermore, PPARγ controls cell proliferation in various other tissues and organs, including colon, breast, prostate, and bladder, and dysregulation of PPARγ signaling is linked to tumor development in these organs. Recent studies have shed new light on PPARγ (dys)function in these three biological settings, showing unified and diverse mechanisms of action. Classical transactivation—where PPARγ activates genes upon binding to PPAR response elements as a heterodimer with RXRα—is important in all three settings, as underscored by natural loss-of-function mutations in FPLD3 and loss- and gain-of-function mutations in tumors. Transrepression—where PPARγ alters gene expression independent of DNA binding—is particularly relevant in immune cells. Interestingly, gene translocations resulting in fusion of PPARγ with other gene products, which are unique to specific carcinomas, present a third mode of action, as they potentially alter PPARγ’s target gene profile. Improved understanding of the molecular mechanism underlying PPARγ activity in the complex regulatory networks in metabolism, cancer, and inflammation may help to define novel potential therapeutic strategies for prevention and treatment of obesity, diabetes, or cancer. PPARG gene on chromosome 3p25.2 in humans  encoding for two protein isoforms via differential promoter usage and alternative splicing . The AF-1 domain is implicated in the constitutive ligand-independent activation of PPARγ target genes. Juxtaposed to the AF-1 domains is the DNA binding domain (DBD) that contains two zinc fingers required for DNA binding. The DBD connected to the ligand binding domain (LBD) via a flexible hinge region. In the case of PPARγ, this hinge region physically interacts with the DNA (PPARγ is a representative member of the nuclear receptor (NR) superfamily. To date, 48 NRs have been identified in human. NRs regulate various critical aspects in development, physiology, reproduction, and homeostasis. NRs are multi-domain ligand-inducible transcription factors that share a structural homology to a varying extent ( the DNA . The ligvia transactivation and transrepression . Transactivation involves a mechanism by which PPARγ binds as a heterodimer complex with RXRα to PPAR response elements (PPREs)  . PPREs c (PPREs) , which a (PPREs) . NoteworIn the last decade, genome-wide binding profiles of PPARγ have been mapped in different cell types, including adipocytes and macrophages –17. ThesTranscriptional control of the target genes by PPARγ furthermore depends on multiprotein coregulatory complexes that are recruited to the PPREs . In basaInterestingly, the transrepression mechanism described above involves a specific post-translational modification, SUMOylation of lysine 365. In fact, to adequately processes external signals and adapt to relevant gene expression programs PPARγ activity is regulated by several, probably interconnected, post-translational modifications, including phosphorylation, acetylation, and the aforementioned SUMOylation [reviewed in ]. Dependin vivo activated by a specific, high-affinity, and endogenous ligand. PPARγ LBD crystal structures reveal a large ligand binding pocket (LBP), which not only allows for promiscuous binding of ligands with lower affinity, but also allows ligands to occupy the canonical LBP in different conformations , a class of anti-hyperglycaemic drugs, including rosiglitazone and pioglitazone . TZDs stWhereas TZDs are commonly referred to as full classical PPARγ agonists, TZDs have a separate biochemical activity: inhibition of the Cdk5-mediated phosphorylation of PPARγ at serine residue 273 . Phosphovia stabilization of helix 3. Furthermore, coregulator-binding assays indicate that alternate site binding has an impact on coregulator interactions, transactivation, and target gene expression , like the aforementioned compound MRL24, and SR1664, can also bind to an alternate site of PPARγ . TZDs, ipression . The idencreased .via “classical” transactivation of target genes.White, beige, and brown adipocytes have been identified in mammals. Although these three type of adipocytes rise from different precursors and differ significantly in their morphology and function, the cells all go through a well-orchestrated differentiation process to become mature and fully functional . During in vivo regulation of adipocyte development, including the stem cell commitment toward white adipocytes, is poorly understood. Adipocyte-lineage tracing, which so far can only be performed in mice, indicate that white adipocytes can be derived from both Myf5− and Myf5+ precursor cells , fatty acid transport , and gluconeogenesis .White adipose tissue (WAT) is the most abundant adipose tissue in the human body . Mature or cells . The Myfasticity . In miceasticity . Althougasticity . After sasticity . PPARγ pPPARG gene . Patienth FPLD3  an increiewed in ]. The FPiewed in Figure 2iewed in .Taken together, genetic mouse models together with the FPLD3-associated PPARγ mutations indicate that PPARγ plays a key role in white AT differentiation, function, and maintenance. The dominant mode of action in this biological setting appears to be “classical” transactivation: the majority of genes regulated by PPARγ in white adipocytes rely on direct DNA binding, and FPLD3-associated PPARγ mutations do not alter transrepression, although this is not studied frequently .UCP1, which encodes for uncoupling protein 1 (UCP1), located in the inner mitochondrial membrane. When activated, UCP1 mediates non-shivering thermogenesis by uncoupling of the oxidative phosphorylation from ATP synthesis, thereby provoking 1) dissipation of chemical energy in the form of heat and 2) stimulating high levels of fatty acid oxidation (Brown adipose tissue (BAT) emerged approximately 150 million years ago in mammals . BAT is xidation .BAT is present in dedicated depots. In rodents, BAT is abundantly present throughout life. In human adults, BAT is located mainly cervical/axillary, perirenal/adrenal, and in the mediastinum along large blood vessels, trachea, and surrounding the intercostal arteries . In new-in vitro cultures  adipocytes . Beige a or both .Although, the exact mechanism by which PPARγ agonists induce browning of white adipocytes is not exactly known, PPARγ agonist require full agonism to activate the browning fat program. The effect is at least in part mediated by PRDM16, a factor that as described above is essential in the development of classical brown fat . TherefoEven though PPARγ is the master regulator of adipocyte differentiation and function , alreadyAs described above for adipocytes, PPARγ plays a role in determining the cellular phenotype by regulating differentiation (adipogenesis) and function  by directly activating the transcription of so-called PPARγ target genes. Similar molecular mechanisms are in place in immune cells, and also here PPARγ can deterimine cellular phenotype: amongst others, PPARγ 1) regulates macrophage differentiation, 2) regulates classical/alternative macrophage activation (“polarization”), 3) controls lipid metabolism in multiple immune cell types, and 4) plays an immune-modulatory role. PPARγ function in immune cells could also be categorized according to its mechanism of action, with the regulation of lipid metabolism and the ability to induce differentiation of immune cells more linked to “classical” transactivation, while the transrepression activity of PPARγ is more important in its immunomodulatory role and both mechanisms are involved in macrophage activation.PPARγ can directly activate the transcription of target genes in immune cells through direct DNA binding, similar to its activity in adipocytes described above. As mentioned earlier, the genomic locations where PPARγ binds and the target genes partly overlap between, for example, adipocytes and macrophages, but cell-type specific regulation may depend on cooperation with other transcription factors like PU.1 and STAT6 , 83.PU1 specifically on monocyte-unique target genes  and secretory output of macrophages, dendritic cells, and T cells. Findings in T cells appear sometimes conflicting, which makes it difficult to assign a clear pro-Th1 or pro-Th2 role to PPARy activation. It also indicates that the use of ligands in these cells might “hide” some of the PPARγ functions and more subtle approaches, such as the use of cels harboring specific PPARγ mutations or selective PPARγ modulators, must be used in order to fully elucidated PPARγ role in immune cells, taking the complex interactions between immune cell population into account.Cancer is driven by the acquisition of genome instability. The cancer genome landscape contains an enormous diverse repertoire of amplifications, deletions, inversions, translocations, point mutations, loss of heterozygosity, and epigenetic changes that collectively result in tumorigenesis. The role of PPARγ in tumorigenesis is controversial. A large body of evidence suggests that PPARγ functions as a tumor suppressor, as activation of the PPARγ/RXRα signaling pathway in different types of cancer, including colon , lung 1, 115, paFurthermore, in lung cancer cells, a tumor suppressive function of PPARγ was contributed metabolic reprogramming , an essePPARG mutations have been reported in ~8% of sporadic colorectal cancers  has been identified. These RXRα mutations rely on the introduction of an aromatic amino acid residue that enhances the ligand-independent activation of PPARγ . Tumors nd FABP4 . In addiRecently, seven recurrent driver gain-of-function PPARγ mutations have been identified in luminal bladder tumors  Figure 2Although, not all recently identified gain-of-function PPARγ mutants have extensively been characterized and even affect different domains in the protein, at least some of the mutants have implications for “classical” transactivation of PPARγ target genes in bladder cancer.PAX8/PPARG fusion gene that is detected in approximately 35% of FTCs and in a subset of follicular variant of papillary thyroid carcinomas (PAX8) encodes for a member of the paired box (PAX) family of transcription factors and is a critical regulator in physiological thyroid development  by acting as a negative inhibitor of tumor suppressor PPARγ or as ii) a novel transcriptional factor with proto-oncogene activity. Nevertheless, the expression of PAX8-PPARG in FTCs does not affect prognosis . The t chromosomal translocation results in a rcinomas . This chrcinomas . The genelopment . In addielopment , 139. Th thyroid . The traf PPARγ1 . The PAXf PPARγ1 . In vitrrognosis .CREB3L2/PPARG fusion gene, is a low incidence fusion mutation that is found in <3% of the FTCs (CREB1L2) encodes for a member of the bZIP transcription factor family. The CREB3L2/PPARγ fusion protein consists of amino acids 1 to 106 of wildtype CREB3L2, a new glutamic acid at position 107 juxtaposed to the all 477 amino acids of wildtype PPARγ1  resulting in a the FTCs . The gene PPARγ1 . The CREe PPARγ1 . The fuse PPARγ1 . The impe PPARγ1 . The once PPARγ1 .Taken together, the PPARγ fusion proteins display a third mode of PPARγ action, as they potentially alter the target gene profile of both parent proteins in the chimeric protein , some of the anti-cancer effects may actually occur indirectly through adipocytes surrounding the tumor or distal adipose tissue. PPARγ plays a crucial role in AT, and as it has been shown before, AT influences cancer initiation and progression through several mechanisms . It is eA second exciting option to consider when considering the use of PPARγ ligands in cancer treatment is the role of the receptor in epithelial to mesenchymal transition (EMT). Epithelial cells that undergo EMT in the primary tumor acquire crucial features that increase their invasiveness, migratory phenotype, and resistance to apoptosis that are essential for the development of metastasis . TransdiPPARγ plays a pivotal role in the crossroad between obesity, immunity, and cancer. Understanding the common and unique molecular mechanism underlying the function of PPARγ in these situations will allow the development of new therapies. In order to do so, some challenges have to be overcome; achieving a selective modulation of PPARγ and a cell-specific delivery of these modulators are two of them. In order to maximize the beneficial effects of targeting PPARγ, the key might be that PPARγ has to be targeted in one specific cell type, and not indiscriminately throughout the whole body. The use of nanoparticles coupled to biological ligands that binds to specific membrane receptors for drug delivery is a technique that is been study for cancer treatment and it could have a bright future in the nuclear receptor field if its proven successful. Given the different and complex roles of PPARy in metabolism, immunity, and cancer, which rely on overlapping and diverse mechsmisms of action, cell-specific delivery of PPARγ ligands, especially noncanonical agonist ligands (NALs) and selective PPARy modulators (SPPARMs), represent a promising field of study for future research.MH-Q, MB, and EK drafted, edited, and revised the manuscript. All authors contributed to the article and approved the submitted version.MH-Q was funded by the European Training Networks Project (H2020-MSCA-ITN-308 2016 721532).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The life-threatening emergency (LTE) in urban and rural areas is an important challenge, which can affect pre-hospital mortality rates. Therefore, non-hospital centers must be prepared to deal with such emergency cases that may occur in the geographic area of activity of these centers. The aim of this study was to identify prevalent domains related to the preparedness of non-hospital centers to deal with the LTE using of healthcare providers’ and experts’ perspectives. A qualitative exploratory study was applied using Semi-Structured Interviews (SSIs) and Focus Group Discussions (FGDs). Prior to beginning on data collection, the study and its aims were explained to participants and their informed consents were obtained. Then SSIs and FGDs were conducted by two trained researchers using an interview guide, which was developed through literature review and consulting experts. In total, 12 SSIs were carried out with 4 clinical officers, 2 nurses and 6 physicians at different non-hospital centers in Tabriz. In addition, 2 FGDs were conducted with specialists in emergency medicine, primary health care, and executives of health centers, with over 5 years of work experience. Purposive sampling method was used as tool to select participants who have sufficient knowledge in our research scope. All SSIs and FGDs were transcribed and immediately recorded word by word. Framework Analysis was employed to manually analyze the interview transcripts from all the SSIs and FGDs.  The interview transcripts analysis resulted in the emergence of 3 domains and 11 sub-domains, categorized according to Donabedian’s triple model. 3 sub-domains were related to inputs, including fundamental infrastructures, support facilities, and human resources. 6 sub-domains were related to processes, including clinical interventions, maintenance of equipment, medicine storage capability, educating and training, infection control, and quality control. Finally, 1 sub-domain was related to outcome, which was also patient’s satisfaction.  Existence of an efficient and effective assessment system can play a pivotal role in identifying the capabilities of non-hospital centers. Therefore, the results of this study can assist health managers to assess the preparedness of these centers.Non-hospital Health centers, Preparedness, Assess, Life-threatening emergency


Strawberries are one of the most economically important berry fruits worldwide and exhibit colours ranging from white to dark red, providing a rich genetic resource for strawberry quality improvement. In the present study, we conducted transcriptome analyses of three strawberry cultivars, namely, ‘Benihoppe’, ‘Xiaobai’, and ‘Snow White’, and compared their gene expression profiles. Among the high-quality sequences, 5,049 and 53,200 differentially expressed genes (DEGs) were obtained when comparing the diploid and octoploid strawberry genomes and analysed to identify anthocyanin-related candidate genes. Sixty-five DEGs in the diploid genome (transcriptome data compared to the diploid strawberry genome) and 317 DEGs in the octoploid genome (transcriptome data compared to the octoploid strawberry genome) were identified among the three cultivars. Among these DEGs, 19 and 70 anthocyanin pathway genes, six and 42 sugar pathway genes, 23 and 101 hormone pathway genes, and 17 and 104 transcription factors in the diploid and octoploid genomes, respectively, correlated positively or negatively with the anthocyanin accumulation observed among the three cultivars. Real-time qPCR analysis of nine candidate genes showed a good correlation with the transcriptome data. For example, the expression of PAL was higher in ‘Benihoppe’ and ‘Xiaobai’ than in ‘Snow White’, consistent with the RNA-seq data. Thus, the RNA-seq data and candidate DEGs identified in the present study provide a sound basis for further studies of strawberry fruit colour formation. Fragaria × ananassa) is an important horticultural crop that has excellent commercial and nutritional value and an attractive appearance. The colour of strawberries results from the accumulation of anthocyanins, with colourful fruit appealing to consumers because of their attractive appearance (PAL) (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3′-hydroxylase (F3′H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS) (UFGT) (GST)  , chalconse (ANS) , UDP glu) (UFGT)  and glutT) (GST) . In Arab and NAC . Many st and NAC . For exa and NAC . RAP encry fruit . F3H is contents . F. ananwberries . FaF3′H t period . InteresANS gene .DFR, leucoanthocyanidin dioxygenase (LDOX), and UFGT by several hundred-fold. In contrast, the expression of CHI, CHS, and C4H (cinnamate-4-hydroxylase) decreases during anthocyanin biosynthesis  were shown to be downregulated in strawberries with a yellow pigment phenotype. In addition, MYB1R expression was shown to be upregulated in strawberries with the yellow pigment phenotype, indicating that MYB TFs repress or enhance anthocyanin accumulation in wild strawberry  and FabHLH3 Δ (which encodes a putative negative regulator) interacts with MYBs to regulate proanthocyanidin biosynthesis in strawberries  are essential regulators for the expression of structural genes in the anthocyanin biosynthesis pathway, such as MYB , bHLH H, WRKY D, and ERFrawberry . F. ananry fruit . In addicessions . Sequencerrensis . Moreovewberries . Accordiwberries . Furtherthaliana . It is ithaliana , while ithaliana .F. ananassa Duch. ‘Benihoppe’), ‘Xiaobai’ (F. ananassa Duch. ‘Xiaobai’), and ‘Snow White’ (F. ananassa ‘Snow White’). The ‘Benihoppe’ strawberry cultivar (cultivated in Japan) as red fruit skin and flesh; the ‘Benihoppe’ cultivar is a white-fleshed mutant of ‘Xiaobai’ that has red fruit skin and white flesh; and the ‘Snow White’ cultivar (cultivated in China) has white fruit skin and flesh  were used in the present study –1I. PlanPRJNA552213).According to a previous study , the devFragaria vesca diploid strawberry genome  database (https://www.ncbi.nlm.nih.gov/genome/3314), abbreviated as in the diploid genome) and the Fragaria x ananassa Camarosa genome , abbreviated as in the octoploid genome) to identify all DEGs in S2 vs. S5, S2 vs. S7, S5 vs. S7 stage comparisons for the ‘Benihoppe’, ‘Xiaobai’, and ‘Snow White’ cultivars, respectively.Reads from each library were assembled separately. Raw sequence reads were filtered using the Illumina pipeline according to the methods of BerryGenomics, Beijing, China. Briefly, after removing the adaptor sequences and low-quality sequences (including the reads with percentages of N over 10%), the remaining high-quality clean reads were used for analysis –S3. The p-value. A log2 value (one sample/one sample) of >1 or <-1 was the criterion used to select candidate genes (anthocyanin biosynthesis pathway (The programs Cuffquant and Cuffnorm used fragments per kilobase of transcript per million fragments mapped (FRKM) as a measure of transcript or gene expression levels . DEseq2  pathway , anthocy pathway  and TFs  pathway  . The cycling program was as follows: 95 °C for 5 min followed by 45 cycles of 95 °C for 30 s, 60 °C for 30 s, and 72 °C for 30 s, which was followed by a melting curve analysis at 60−95 °C and obtained one melting point then subsequently used for further study. The relative expression level of each gene was calculated using the comparative 2−ΔΔCT method  using an E.Z.N.A. Plant RNA Kit following the manufacturer’s instructions. The concentration and quality of RNA were analysed on a NanoDrop 1000. Approximately 1.5 µg of total RNA was used for cDNA synthesis using a PrimerScriptTM RT Reagent Kit with gDNA Eraser . All the primers used in the qRT-PCR analysis were designed using Vector NTI  without T method . All anaFor anthocyanin extraction, 5 g of frozen powders of fruits from three stages  for the three strawberry cultivars were used in the present study . To extrSubsequently, 5 g of frozen powders of fruits from three stages  for the three strawberry cultivars were used for soluble sugar analysis . The powEach experiment was independently repeated at least three times. The data were statistically analysed with a one-way analysis of variance of Duncan’s multiple range text using IBM SPSS Statistics 20. The results are presented as the mean value ± standard deviation of the mean (SD) ; Fig. 6,In the present study, the anthocyanin composition and contents of the red-fruited strawberry (‘Benihoppe’), the white-fleshed mutant of ‘Benihoppe’ (‘Xiaobai’), and the white-fruited strawberry (‘Snow White’) were analysed –1I. As ePRJNA552213). On average, 91.2% of the reads from the 18 libraries had a quality score over Q30. Moreover, over 99% and approximately 90% of the cleaned reads could be uniquely mapped to the diploid and octoploid strawberry genomes, respectively. Pearson’s correlation coefficients of the transcriptome profiles were 0.99 between each set of biological replicates, indicating that the sequencing quality was suitable for further analysis  of ‘Benihoppe’, ‘Xiaobai’, and ‘Snow White’ with two repeats were used for sequence analysis with the Illumina HiSeq platform. These libraries were subjected to RNA-seq using an Illumina HiSeq X Ten, generating 27,673,862, 33,937,779, 27,331,497, 28,914,712, 32,588,549, 26,390,260, 25,061,869, 29,690,765, 25,703,115, 26,179,018, 26,487,231, 28,493,685, 25,410,599, 31,471,955, 30,928,839, 28,521,409, 28,248,711, and 25,042,090 150-bp paired-end raw reads . All of analysis , S3.Using the diploid strawberry genome database, 10,354 DEGs were identified by comparing our transcriptome data with the diploid strawberry genome. A Venn diagram showing the DEGs from the three strawberry cultivars (in the diploid and octoploid genomes) was drawn using TBtools . In totaPAL (gene 22493), F3H (gene 1350), DFR  and 3-GT (gene 25816 and gene 7173). One DEG (gene8111) involved in the anthocyanin biosynthesis pathway was upregulated in the fruits of ‘Snow White’ and ‘Xiaobai’ compared to those of ‘Benihoppe’, while the expression levels of these genes in ‘Snow White’ were similar to those observed in ‘Xiaobai’. In addition, the expression levels of eight structural genes in the octoploid strawberry, namely, PAL, CHS, CHI, DFR, F3H, ANS, UFGT, and 3-GT, were lower in ‘Snow White’ than in ‘Benihoppe’ and ‘Xiaobai’. These results suggest the involvement of these genes in the anthocyanin biosynthesis pathway, where enhancing or repressing their expression possibly blocks anthocyanin biosynthesis in ‘Xiaobai’ and ‘Snow White’.To investigate which anthocyanin synthesis step was blocked in ‘Xiaobai’ and ‘Snow White’, we compared the expression of anthocyanin-related genes in three developmental stages S2, S5, and S7) of ‘Benihoppe’, ‘Xiaobai’, and ‘Snow White’ to test whether the expression of some of anthocyanin-related genes affect fruit colour. Twelve DEGs were identified in the anthocyanin biosynthesis pathway among the three cultivars in the diploid genome (as mentioned above) , S5, of , and S7 PAL (nine genes in the octoploid genome chromosome 6 (three) and chromosome 7 (six)). The expression profiles showed that PAL (one gene in the diploid genome and five genes in the octoploid genome) was highly expressed in ‘Benihoppe’ and ‘Xiaobai’ compared to ‘Snow White’ during the fruit development and ripening process. The expression of four genes encoding PAL in the octoploid genome showed no major differences among the three cultivars (DFR (DFR gene located at chromosome 3 was extremely low compared to that located at chromosome 2 among the three cultivars in the octoploid genome. Gene 20514 (homologous gene augustus_masked-Fvb6-4-processed-gene-148.10 in the octoploid genome), which was predicted to encode a polyphenol oxidase gene involved in the pigment biosynthetic process, was upregulated during the fruit development and ripening process in ‘Benihoppe’ and ‘Xiaobai’ compared to that observed in ‘Snow White’ (over 50-fold higher) (UGT 91A1) involved in the flavonoid biosynthetic process, was upregulated during the fruit development and ripening process in ‘Benihoppe’ and ‘Xiaobai’ compared to that observed ‘Snow White’ (over 2-fold higher)  , S5. Pol higher) . Moreove higher) , S5. TheTo assess the association between sugar and fruit colour, we measured the contents of three sugars and analysed the expression profiles of DEGs involved in sugar-related genes according to a GO enrichment analysis , S5. ThePLT 5-like)) have the same annotation and exhibited similar expression profiles between the octoploid and diploid genomes. It is interesting to note that the bidirectional sugar transporter is present in both the diploid  and octoploid  genomes, and their expressions was upregulated in ‘Benihoppe’ and ‘Xiaobai’ (approximately 10-fold higher expression levels than in ‘Snow White’) compared to ‘Snow White’ (PLT 5-like expression was downregulated during the fruit development and ripening process in ‘Benihoppe’ and ‘Xiaobai’ compared to that observed in ‘Snow White’ (over 3-fold higher)  , S5. ThePP2C 37), and gibberellin 2-beta-dioxygenase (GA2Ox) have the same annotation and expression profiles in the octoploid and diploid genomes. Among these factors, auxin-induced protein 15a-like was encoded by one gene (gene 4393) in the diploid genome and seven genes in the octoploid genome, which were located at chromosomes 2 and 5, respectively  in the diploid genome (PP2C had three geneIDs and was located at chromosome 7 in the octoploid genome (PP2C 37 genes were more highly expressed in ‘Benihoppe’ and ‘Xiaobai’ compared to ‘Snow White’ (approximately 20-fold higher). In contrast, only one gene (PP2C 37) was more highly expressed in ‘Snow White’ than in ‘Benihoppe’ and ‘Xiaobai’ (approximately 10-fold higher). With respect to GA2Ox, one gene (gene 7769 in the diploid genome) and four genes (in the octoploid genome) were upregulated during the fruit development and ripening process in ‘Benihoppe’ and ‘Xiaobai’ compared to that observed in ‘Snow White’ (two genes)  , S5. In bHLH130, MADS23, and WRKY22) were identified as having the same annotations in the diploid and octoploid genomes. The expression pattern of bHLH130 and MADS23 was higher in ‘Snow White’ compared to that observed in ‘Benihoppe’ and ‘Xiaobai’ and contrasted with the pattern of anthocyanin accumulation observed in the three cultivars  and MYB44 on chromosome 1 were more highly expressed than MYB44 on chromosome 2 and 5. MYB44, which is located on chromosome 1, was upregulated in red fruit compared to white fruit. MYB44 located on chromosomes 2 and 5 was downregulated in red fruit compared to white fruit . In addition, the expression patterns of FaPAL1, FaF3H, FaDFR, FabHLH, and FaTT12 were observed to be consistent with their expression profiles in diploid and octoploid genomes. The qRT-PCR results were similar to our RNA-seq results, supporting the reliability of our RNA-seq data.To validate the RNA-seq-based DEG data, we quantified the expression of nine DEGs related to anthocyanin biosynthesis using quantitative real-time PCR (qRT-PCR) for the ‘Benihoppe’, ‘Xiaobai’, and ‘Snow White’ cultivars . All selStrawberry, an important horticultural crop worldwide, has excellent commercial and nutritional value and benefits human health . AnthocyCHS, CHI, and ANS, which were not identified in the diploid genome but were present in the octoploid genome, were downregulated in white strawberry fruits, indicating that the transcript abundance of these genes was positively related to the accumulation of anthocyanin , while some copies were expressed at low levels . The expression profiles of DFR were consistent with a reduction in anthocyanin accumulation and a yellow pigment phenotype (F3′H was not identified in the present study (no significant difference between two genomes), with similar results having been obtained in previous reports  was downregulated during the strawberry fruit ripening process, the expression of which contrasted with that of UFGT, which enhances cyanidin glucoside accumulation  in our study indicated that this gene is involved in anthocyanin biosynthesis  has been shown to play important roles by interacting with MYB during proanthocyanidin and anthocyanin biosynthesis  was upregulated in white fruit compared to red fruit and may be negatively regulated anthocyanin biosynthesis by competitively consuming substance of anthocyanin. In addition, some bHLHs have also been shown to be responsible for fruit anthocyanin biosynthesis according to their expression profiles in three octoploid strawberry cultivars  plays a crucial role in controlling anthocyanin biosynthesis by acting directly or indirectly through MYB transcription factors to control carbon flux through the phenylpropanoid pathway  and two structural genes (AT1G68440 and AtGSTF12) involved in anthocyanin biosynthesis  and 53 DEGs (in the octoploid genome) having the same annotation between the diploid and octoploid genomes) that may be associated with anthocyanin biosynthesis in white and red strawberry cultivars. Overall, the results of the present study provide insights into the possible molecular mechanisms influencing strawberry fruit colour, which is of great importance for both basic research and the breeding of strawberry for white fruit.10.7717/peerj.10739/supp-1Supplemental Information 1Table S1. The qulity of transcriptome sequencing data statistics.Fragaria vesca.Table S2. Pearson’s correlation coefficients between biological replicates. Of the transcriptome profiles obtained by comparing with Fragaria x ananassa.Table S3. Pearson’s correlation coefficients between biological replicates. Of the transcriptome profiles obtained by comparing with Fragaria vesca. Differentially expressed genes  >1 or < − 1 was the selection criteria) of the anthocyanin-related pathway, sugar-related pathway, hormonal pathways and transcription factors. The number indicated candidate gene FPKM.Table S4. Expression patterns of candicate genes in Fragaria x ananassa. Differentially expressed genes  >1 or < − 1 was the selection criteria) of the anthocyanin-related pathway, sugar-related pathway, hormonal pathways and transcription factors. The number indicated candidate gene FPKM.Table S5. Expression patterns of candicate genes in Table S6. The primers names and sequences used in this study.g−1 FW).Table S7. The content of anthocyanins in three cultivated strawberry. The number indicated anthocyanins content (µg−1 FW).Table S8. The content of sugars in three cultivated strawberry. The number indicated sugars content (mg kgTable S9. Expression levels of anthocyanin pathway genes by qRT-PCR. The number indicated candidate gene expression levels.Click here for additional data file.10.7717/peerj.10739/supp-2Supplemental Information 2Click here for additional data file.