The lowest SECs for the desalination of SGW with a TDS concentration ≥2 g/L were determined by the use of BWRO and SWRO with 0.4–2.9 kWh/m3 for a water recovery of 40–66%. Even though the MCDI technique cannot compete with pressure-driven membrane desalination technologies at higher raw water salinities, this technology shows a high selectivity for nitrate and a high potential for flexible desalination applications.Saline groundwater (SGW) is an alternative water resource. However, the concentration of sodium, chloride, sulphate, and nitrate in SGW usually exceeds the recommended guideline values for drinking water and irrigation. In this study, the partial desalination performance of three different concentrated SGWs were examined by pressure-driven membrane desalination technologies: nanofiltration (NF), brackish water reverse osmosis (BWRO), and seawater reverse osmosis (SWRO); in addition to one electrochemical-driven desalination technology: membrane capacitive deionisation (MCDI). The desalination performance was evaluated using the specific energy consumption (SEC) and water recovery, determined by experiments and simulations. The experimental results of this study show that the SEC for the desalination of SGW with a total dissolved solid (TDS) concentration of 1 g/L by MCDI and NF is similar and ranges between 0.2–0.4 kWh/m Due to a continuously growing population, economic development, and changing consumption patterns, potable water consumption has increased globally. At the same time, the quality of groundwater and surface water has decreased, causing a reduction in the levels of terrestrial water storage and water scarcity in regions such as the Middle East, India, Australia, and Africa ,2,3,4. Sghly SGW ,11, as lural use ,13. Howeural use ,15.3 and for SWRO between 2.5–4.0 kWh/m3.Desalination is a separation process used to reduce the concentration of TDS in saline water. Desalination technologies can be classified by the separation procedure. Either the pure water is removed from the feedwater as is the case in distillation and pressure-driven membrane desalination, or the salts are removed from the feedwater as is done in electrodialysis and capacitive deionisation . At pres3 depending on the feed and target water concentration.Nanofiltration is another cost-effective desalination technique characterized by a looser membrane compared to RO membranes, resulting in a higher permeate flux and a lower rejection of monovalent ions . Accordi3, treating brackish water with a salinity of 1–5 g/L and a water-recovery of 50%.Electrochemical-driven membrane technologies such as electrodialysis and capacitive deionisation (CDI) are alternative flexible desalination technologies for brackish water with a low requirement for chemical additives . ElectroThe steadily increasing demand for freshwater in regions affected by saltwater intrusion is driving the need for energy efficient desalination technologies. The desalination of SGW is—due to the relatively low salinity and low required energy consumption compared to seawater desalination—a vital solution for solving problems related to water stress and scarcity ,36. ManyHence, the objective of this study is to compare the desalination performance of SGW by pressure-driven membrane desalination technologies—using NF, BWRO, and SWRO—and an electrochemical technique—using MCDI—to answer the research question: which technique has the lowest SEC regarding different desalination scenarios? Therefore, the desalination performance in this study is evaluated by the volume-related specific energy consumption  is defined by the ratio of uctivity . Since tlination  as well lination , the SARThe mass concentrations of SGW SECVSince the desalination performances were only evaluated for mixed salt solutions and not for single salt solutions, the The concentration of the synthetic SGW and the target concentration in the product water were default values in this study. The water recovery, however, can vary depending on the operation and system parameters of the used desalination technology. Additionally, a distinction was made between the module-scale water recovery (ion (6)) .(6)γSysThe module-scale salt rejection depends on the properties of the specific pressure-driven membrane desalination and MCDI modules, as well as on the specific process parameters such as pressure, applied electrical current, and flow rate. Therefore, the module-scale salt rejection (In case ) of SGW  to achieg isR the ideal gas constant ) .(8)∆Gmilination .(9)SECm3/h, which represents a small-desalination plant [® Design Software 2.1.1 by LANXESS Deutschland GmbH, Cologne, Germany [The energy-consuming factor for pressure-driven membrane desalination is the feed pressure. The feed pressure needs to be adjusted to ensure the required permeate (and concentrate) flow rate, considering the friction losses and the concentration polarization in a membrane module. Therefore, the feed pressure in a small-scale design differs from that in a big-scale design. In order to calculate the SEC for a realistic large-scale desalination plant, the production flow rate was set to 25 mon plant . The reqon plant . In thison plant . The reqon plant  and Lewa Germany ,60). In The water permeability The net driving pressure results from the pressure difference The ideal desalination membrane has a high water permeability paired with a near-zero salt permeability for ions which need to be rejected . Since bThe required feed pressure for each desalination scenario was set according to Equation (10). Therefore, a single-module design with 6-elements in series was selected. The number of parallel modules was adapted to the active surface area of the specific membrane modules, the required permeate flow, and the module-scale recovery. The module-scale recovery was set to 60% for NF and BWRO, and 40% for SWRO, according to the design guidelines for the specific membranes ,59,62,63In total, the SEC for pressure-driven membrane desalination depends on the water and salt permeability of the membranes, the water temperature, and the salt concentration in the feedwater. The SEC for pressure-driven membrane desalination . The active membrane area was set to 210 mm × 40 mm. Since the height of the feed channel was set to 1.016 mm by the membrane holder, the same diamond shaped spacer with a height of 40 mil was used for all experiments. All experiments were performed using a constant feed pressure at two different pressure stages, which were adjusted to the recommended permeate flux ranges of the membrane manufacturers [The desalination performance was investigated at the Leibniz Institute of Polymer Research Dresden using an OSMO Inspector 2  and a stainless steel membrane holder of SIMA-tecacturers ,59,63. SBefore each desalination test, the membranes were wetted and compacted gradually with deionized water until the permeate flux was stable. Since the retention for highly SGW needed to be higher than 95%, NF was not evaluated for desalination of highly SGW. All desalination tests were performed in triplicate.The permeate flow and the transmembrane pressure were measured online in each test period for 1–2 h. The concentration of single ions, the pH, and the electric conductivity were measured in the feed, concentrate, and permeate sample for each experimental run. The qualitative sample analyses were carried out at the Institute of Urban and Industrial Water Management at the Technische Universität Dresden. The pH and conductivity were measured using a HQ40D portable multi meter from Hach, Düsseldorf, Germany, with a PHC 301 electrode and a CDC 401 electrode. The cations were measured by atomic absorption spectroscopy using a SpectrAA 220FS Varian, Agilent, Palo Alto, CA, USA, according to DIN 38406-3:2002-03  and DIN :2005-12 .The most energy-consuming factor of MCDI is the electric energy needed to generate an electric field between the electrodes. The energy demand of the feed pressure can be neglected, since the feedwater is just flowing by and not pressed through a membrane ,35,71. Isorption .(13)SEC3/h desalinated water using Python 3.7 .The The electrosorption process in CDI can be described by the formation of the electrical double layers within the micropores of the carbon electrodes . In the 2. In this study, we wanted to achieve a constant diluate quality. Therefore, the desalination tests were performed using a constant electrical current [The MCDI small-scale experiments were conducted with a C 03 25 DDRG MCDI module integrated in the CapDi Pilot Unit from 2010 of Voltea B.V., Sassenheim, Netherlands, at the Technische Universität Dresden. The pilot plant was controlled by using the “Demo Unit software” from Voltea B.V. (2010) and Tera Term v. 4.92. The total electrode surface area of the module was 3.7 m current . The durk to 0 V .The required electrical current (on (14)) ,32,77.(According to Equation (14), a higher electrical current was selected in order to achieve the higher required salt retention. However, the MCDI module used in this study is not designed for desalination of highly SGW. Hence, desalination tests were performed only with slightly SGW and moderately SGW. In this study, 10 A, 14 A, 18 A, 22 A and 26 A were applied for desalination of slightly SGW, and 20 A, 40 A, 45 A, 50 A and 55 A for moderately SGW. The feed flow  and 5.91 L/(m2 h bar)  a. The hiand NO3− b.The experimental determined normalized water permeability at 25.0 °C follows the order of slightly SGW > moderately SGW > highly SGW . In gene3, whereas this was 0.4 to 1.5 kWh/m3 for desalination of moderately SGW with BWRO or SWRO  than in the feed (7.3–8.7).The ion selectivity in ectively . The SECIndependent of the method used, the lowest ejection . Therefo for Na+  was margcurrents . As a reIf a higher salt rejection for desalination of slightly SGW is required to comply with the target concentrations of local freshwater, pressure-driven membrane desalination technologies are more advantageous due to their higher system-scale water recovery, even though MCDI showed a similar SECV . AccordiThe laboratory tests were examined under ideal conditions. Even though precipitation was not detected in the concentrate during short-term experiments, the solubility limit was exceeded by simulating big-scale pressure-driven membrane desalination. Other substances, which were not examined in this study, such as silica, iron or organic matter, can further enhance the scaling and fouling potential ,96 and wThe obtained n et al.  and Zhaon et al. . The expterature ,20,21. Dterature  for sligIn this study, the desalination performance was examined for one-stage configurations to ensure similar conditions for pressure-driven membrane desalination and MCDI. According to Werber et al.  and ShriOverall, the experimental and simulated results in this study showed higher SECV  than SECV values  due to im design ,100. Accm design  future rm design . In ordem design ,103,104.For pressure-driven membrane desalination as well, as for MCDI, the feedwater temperature is an important factor influencing the energy efficiency, the SEC, and the water recovery. In this study, the desalination performance was evaluated for the specific temperature of the respective SGW. The dialysis . Howeverdialysis , the salThe module-scale water recovery and the module-scale salt rejection of NF, BWRO, and SWRO were linked to narrow operation guidelines of the module specifications for the flow rate of the feed, permeate, and concentrate. The total flow rates therefore depend on the specific design of the membrane elements. The adjustment of the flow rate for MCDI was, however, flexibly adjustable according to the required salt rejection and the applied electrical current. Therefore, MCDI shows great potential for meeting flexible desalination demands in terms of salt rejection and water recovery . Due to This study analysed the SEC for pressure-driven membrane desalination technologies and MCDI to compare the desalination performance for mixed salt solutions. In order to compare the total costs, further cost parameters, such as the capital costs, the required area for the desalination plant, and the adapted pre- and posttreatment costs need to be considered as well. Moreover, both desalination technologies—pressure-driven membrane desalination and MCDI—can benefit from each other in hybrid processes . Due to In this study, the desalination performance was examined for three different realistic SGW concentrations using NF, BWRO, SWRO, and MCDI. Our experimental results indicate that pressure-driven membrane desalination and MCDI show—depending on the operation procedure—different selectivity towards The lowest SEC values for a low level of required salt rejection, such as for slightly SGW (TDS = 1 g/L), were achieved with MCDI and NF. Thereby, the lowest SEC values were obtained with MCDI for slightly SGW with regard to the guideline concentrations for irrigation and drinking water. However, if a higher salt rejection is required, as for the target concentration for local freshwater, NF demonstrated a better desalination performance than MCDI, due to a higher water recovery. Pressure-driven membrane desalination—such as BWRO and SWRO—demonstrated, independent of the target concentrations under the respective boundary conditions, a better desalination performance for TDS concentrations ≥2 g/L.Even though the experiments of this study confirm that pressure-driven membrane desalination technologies show a higher energy efficiency regarding higher saline concentrations, we showed that MCDI is particularly suitable for desalination of


To investigate the association between different stages of posterior vitreous detachment (PVD) and macular microvasculature in the elderly.Swept-source optical coherence tomography (OCT), OCT angiography, and color fundus images of 490 eyes without retinal pathologies of 322 participants aged ≥65 years were evaluated. PVD was classified using enhanced vitreous visualization mode as no apparent PVD (stage 0/1), vitreous adhesions at the fovea and optic disc (stage 2), adhesion at the optic disc (stage 3), or complete PVD (stage 4). Microvascular parameters, including foveal avascular zone (FAZ) and vessel density (VD), were analyzed for their associations with complete PVD. Additionally, the association between PVD and central retinal thickness (CRT) was also addressed.2; P = 0.014), and the mean superficial VD was lower in stage 4 compared with stages 0–3 . However, PVD was not significantly associated with deep macular microvascular parameters or CRT.Overall, 80, 31, 31, and 349 eyes were categorized into stages 0/1, 2, 3, and 4, respectively. Using multivariate mixed-effects model, the mean superficial FAZ area was smaller in stage 4 compared with stages 0–3 (0.29 vs. 0.32 mmComplete PVD was associated with smaller FAZ area and lower VD in superficial macular microvasculature, while it was not associated with central retinal thickness. Posterior vitreous detachment (PVD) is one of the most common physiological phenomena experienced by the middle-aged and elderly populations. It manifests as vitreous gel liquefaction and weakened vitreoretinal adhesion. It prodOptical coherence tomography angiography (OCTA) is based on motion contrast imaging of high-resolution volumetric blood flow data. It allows for the generation of angiographic image slices of the retina as well as non-invasive three-dimensional vascular mapping of retinal microcirculation at different retinal depths. Recently, OCTA has been evaluated in clinical studies, proving particularly useful in those on retinal diseases. Images The purpose of the current study is to investigate whether or not there is an association between PVD evaluated with SS-OCT and macular microvascular parameters obtained via OCTA among the community-dwelling elderly population in a metropolitan area of Tokyo.This cross-sectional study was conducted in 2018 as part of a prospective cohort study enrolling the community-dwelling elderly population in the Itabashi ward, a northwestern area of Tokyo. The cohort study, which had been conducted since 2011 and called “OTASSHA-KENSHIN” , was approved by the institutional review board and ethics committee of the Tokyo Metropolitan Institute of Gerontology,14, and In the OTASSHA-KENSHIN study, community-dwelling volunteers aged 65 years or older annually underwent comprehensive clinical examinations, including hematology tests, body composition measurements, cognitive function tests, and comorbidity and social background questionnaires. In 2018, in addition to these annual comprehensive examinations, the participants underwent fundus color photography, OCT, and OCTA with experienced optometrists who were not aware of the purpose of the present study.For the current study, the exclusion criteria were as follows: 1) eyes with a history of retinal disease , as reported by the participants through a questionnaire or demonstrated by fundus photos, OCT, or OCTA obtained through the current study; 2) images with insufficient quality, as measured by a built-in software  value of 40 or less); and 3) The participants underwent OCT/OCTA using an SS-OCT  with a wavelength of 1,050 nm, an acquisition speed of 100,000 A-scans per second, and an axial and transversal resolution of 7 and 20 μm, respectively, in tissue. Two trained operators captured standard three-dimensional 4.5 mm × 4.5 mm macular cubes for each eye. Slabs of the superficial capillary plexus (SCP) and deep capillary plexus (DCP) were automated and segmented by the built-in software . The SCP was delineated from 2.6 μm below the internal limiting membrane to 15.6 μm below the junction between the inner plexiform and inner nuclear layers, and the DCP was delineated from 15.6 μm below the inner plexiform and the inner nuclear layers to 70.2 μm below them[The SS-OCT images covered the posterior retina, including the macula and optic nerve disc (7 mm × 7 mm area centered on the fovea). The extent of PVD was graded based on a recently proposed system[CRT has been known to be associated with FAZ area. We addrUsing the ImageJ software , the grayscale OCTA images were standardized and cropped (3 mm × 3 mm), with the fovea at the center.The ImageJ macro for the FAZ measurement was as follows: Each image was edited in the order of smooth Gaussian Blur 3D  and Auto Local Threshold minimum . Then, uFor the VD measurement, using ImageJ, each image was binarized via the Niblack method , and theP-value of <0.05 was regarded as statistically significant.A statistical analysis was performed using the JMP Pro 14 software (SAS). For the univariate analysis, the ANOVA and Fisher's exact test were used to compare continuous and categorical data, respectively, among the different PVD stages. To include both eyes of the same participant in the analysis, a mixed model was applied for the multivariate analysis. The effects of age, sex, diabetes mellitus (DM), hypertension, and smoking status  were adjusted to explore the possible effect of PVD on retinal microvascular parameters. The summarized continuous data were presented as mean ± standard deviation (SD). A Images of 490 eyes (322 participants) were judged eligible for analyses in the present study. Among the 490 eyes included, 80 (16.3%), 30 (6.1%), 31 (6.3%), and 349 (70.9%) were categorized into PVD stages 0/1, 2, 3, and 4, respectively. 2, P = 0.014; P = 0.0089; Next, we investigated the effect PVD on macular microvascular parameters. As the number of eyes with PVD stage 2 or 3 was relatively small, we compared the effects of PVD stages 0–3  with those of stage 4 . After adjusting for the effects of age, sex, and history of DM, hypertension, and smoking status, stage-4 PVD was significantly associated with a smaller superficial FAZ area compared with stages 0–3  with those of stage 4  on macular microvascular parameters. Again, compared with stage 0/1, stage-4 PVD was associated with a smaller superficial FAZ area (0.31 vs. 0.29 mmP = 0.77) or between PVD stage 0 vs. stage 4 . The results did not support the hypothesis that CRT is involved in the effect of PVD on macular microvascular parameters.From the mechanistic viewpoint, we speculated that PVD could influence the macular microvascular parameters through its effect on central retinal thickness (CRT), because CRT is associated with FAZ area in healthy eyes. TherefoThe present study investigated the association between PVD and retinal microvasculature via SS-OCT and SS-OCTA among the community-dwelling elderly in Tokyo. Our results suggest that PVD has effects on superficial retinal microvasculature in the elderly, as demonstrated by a smaller FAZ area and lower macular VD. However, PVD does not appear to have a significant effect on deep macular microvasculature or CRT.In previous studies, correlations of retinal microvasculature to demographic, systemic, and retinal diseases have been extensively studied using OCTA. For example, females have a larger FAZ area in normal eyes. In addiOur conclusion that PVD is associated with a smaller FAZ area and lower VD was consistent across two comparisons, one comparing PVD stages 0–3 with stage 4 and the other comparing stage 0/1 with stage 4. We speculated how PVD could confer such effects on the superficial macular microvasculature. According to a recent report in which PVD development was investigated on montaged SS-OCT images, a subtle separation of the vitreous cortex from some parts of the retina has started  in most of the elderly, and PVDIn addition to physical changes in the tangential force on the retina, PVD is accompanied by autoregulatory physiological changes. Previous reports have revealed that the partial pressure of oxygen in the vitreous increases in the presence of PVD, by whicFurthermore, the change in tangential force and retinal oxygenation induced by PVD is considered to affect the inner retina more than the outer retina, which is consistent with our results that only superficial macular microvascular parameters are influenced by PVD.Several limitations exist in this study. To start, the enrolled participants were community-dwelling elderly volunteers whose medical and social backgrounds were not uniform. We performed multivariate analyses adjusted for age, sex, history of DM and HT, and smoking status to carefully address the possible effects of PVD on retinal microvasculature evaluated via SS-OCT. However, current understanding about the factors affecting OCTA-imaged retinal microvasculature is imperfect, and we could not adjust for unexpected confounding factors. In addition, the present study was not performed based on detailed ophthalmic examinations; medical history was collected through questionnaires, and fundus color, OCT, and OCTA images were carefully examined to detect pre-existing retinal pathologies. We could not collect data on refractive error or axial length, which might have caused some errors in the FAZ area estimates. HoweverIn summary, the association between PVD and macular microvasculature imaged via SS-OCT and SS-OCTA was investigated in the community-dwelling elderly in Tokyo. We observed a consistent association between complete PVD with a smaller FAZ area and lower VD in the superficial macula but not in the deep macular microvasculature. Our data, strengthened by the inclusion of 490 eyes, suggest an impact of PVD on macular microvasculature in the elderly.


Streptomyces and results in serious losses to the global economy every year. Via high-throughput sequencing, it was seen that the composition of endophytes in roots, stems, and tubers had significant differences (P < 0.05) and the distribution of the bacterial communities illustrated a gradient from soil to root to tuber/stem. PCS significantly reduced bacterial endophytes α-diversity indexes, including ACE and the number of observed operational taxonomic units (OTUs), of RE without significantly reducing the indexes of SE and TE. No significant effect on the composition of endophytes were caused by PCS in roots, tubers, or stems between high PCS severity (H) and low PCS severity (L) infections at the community level, but PCS did have a substantial impact on the relative abundance of several specific endophytes. Rhizobium and Sphingopyxis were significantly enriched in root endophytes with low PCS severity (REL); Delftia and Ochrobactrum were significantly enriched in stem endophytes with low PCS severity (SEL); Pedobacter, Delftia, and Asticcacaulis were significantly enriched in tuber endophytes with high PCS severity (TEH). OTU62, a potential PCS pathogen in this study, was capable of colonizing potato tubers, roots, and stems with few or no symptoms present. Co-occurrence networks showed that the number of correlations to OTU62 was higher than average in these three tissue types, suggesting the importance of OTU62 in endophytic communities. This study clarified the distribution and composition of potato endophytes in tubers, roots, and stems, and demonstrated the response of endophytes in non-lesion tissues to PCS.The response of plant endophytes to disease within infected tissues has been well demonstrated, but the corresponding response of endophytes in non-lesion tissues remains unclear. Here, we studied the composition and distribution of bacterial endophytes in potato roots (RE), stems (SE), and tubers (TE), and explored the response of endophytes in non-lesion tissues to potato common scab (PCS), which is a soil-borne disease caused by pathogenic  Endophytes are bacteria and fungi that exist saprophytically in plants, many are parasitic or symbiotic and live in healthy plant tissues at certain or at all life stages but, generally, do not cause damage to plant health. Endophytes are widely distributed in roots, stems, leaves, seeds, other tissues, and intercellular spaces of various plants . VariousRhizobium could colonize the root surface and then enter the interior of rice roots, becoming endophytic before rising to the stem, leaf sheath, and finally leaf. The diversity and abundance of endophytes in the root of reeds were higher than those of in the above-ground tissues  is considered to be one of the largest soil-borne diseases in the potato market, causing serious economic losses around the world. PCS causes shallow and deep corky blemishes on the surface of potatoes with the phytotoxin thaxtomin  were cut and collected after the leaves were removed. Tubers and roots were carefully collected with aseptic stainless-steel shovels, and the soil that was loosely attached to the tubers and roots was gently removed. All samples were transported to the laboratory within 12 h under low temperature conditions (4°C). GS and RS were classified as soils tightly attached to the surface (about 1 mm) of tubers and roots, respectively. After soil sampling, the roots, stems, and tubers were washed three times with sterile, distilled water, then soaked in 70% ethanol for 2 min, followed by sterile water twice, then soaked in 3% sodium hypochlorite twice, for 1 min each, and finally washed with sterile water three times at 2 min each. We subsequently used the last sterile water rinse to inoculate a control plate to check the disinfection process. All the plant tissue was then cut into small pieces, ground into a homogenate, and stored at −80°C as preparation for DNA extraction.TM Plant DNA Kit  to extract DNA from the three kinds of potato tissue samples according to the manufacturer’s instructions. We measured the concentration and quality of DNA with a NanoDrop 2000 spectrophotometer , and all DNA samples were stored at −80°C until further analysis.We used the E.N.Z.A.PRJNA657530.The sequencing of endophyte communities and soil communities were carried out in a single batch with the same amplification primers, libraries construction, and sequencing conditions. The raw sequence data for the endophytic communities is publicly available under Bioproject number ® UltraTM DNA Library Prep Kit for Illumina , and index codes were added. Amplicon libraries were sequenced on a MiSeq PE250 sequencer  with 250–bp paired–end read. To optimize data quality, we used USEARCH v. 9.2 software to merge pairs, remove primers, and connectors, and filter out low-quality and short sequences. Then, we removed chimeras and generated operational taxonomic units (OTUs) at 97% similarity. We generated an OTU profile for each of our 50 samples according to the RDP database, which included about 34,621 sequences per sample. We then removed any OTUs assigned as chloroplasts or mitochondria and counted the relative abundance of taxonomic profiles at the level of phylum and genus.The primers 341F (5′–CCTACGGGNGGCWGCAG–3′) and 805R (5′–GACTACHVGGGTATCTAATCC–3′) were used to amplify the V3–V4 hypervariable region of the 16S rRNA gene. Per the manufacturer’s instruction, amplicon libraries were constructed using the NEB Nextdiversity function from the “vegan” package in R 3.6.3.Based on the OTU profile, we calculated α-diversity indices, including ACE, Chao1, and Obs (the number of observed OTUs) of endophytes and soil bacteria communities using the anosim function from the “vegan” package in R with the weighted unifrac matrix.We used a repeated-measures design, where microbial sampling was performed on the same individuals but in different tissues, so we used a multivariate approach  to discern subtle differences in the bacterial community composition between different tissue types and to eliminate individual variation. We performed sPLSDA prior to rarefaction . sPLSDA 2. We used the following R packages to organize, analyze, and visualize the data: package “vegan,” “dplyr,” “doParallel,” “foreach,” “mgcv,” “reshape2,” “ggplot2,” “cowplot,” “Rcpp,” and “RcppArmadillo.” We traced RE from GS, RS, and SE, SE from GS, RS, and RE, and TE from GS, RS, SE, and RE. We expressed the sources of different samples as percentages of the total contribution, which were calculated from the average of the FEAST results per sample.We traced the source of endophytes of each plant tissue sample using Fast Expectation-maximization for Microbial Source Tracking (FEAST) . All scrt-test with the parameters of “Two-sided.”In order to explore the differences in the endophyte communities between different tissues, we used STAMP  to ident3 aligned sequence reads. A phylogenetic tree was calculated by neighbor-joining method and the results of 1,000 bootstrap trials were shown at the nods. The evolutionary distances were computed using the Jukes-Cantor method and are in the units of the number of base substitutions per site.We used Molecular Evolutionary Genetics Analysis (MEGA7) to infer the phylogenetic affiliations and relationships of the strains using NCBI’s Microbes BLASTrcorr” function from “Hmisc” package in R, and kept the correlations of P < 0.05 to construct the network. The network was visualized using Gephi.The co-occurrence network was constructed to assess the importance of OTU62 in the endophytic community. We calculated the Pearson’s correlation coefficient between OTUs via the “P < 0.05 was considered significant.For all statistical analyses, N = 1,202) bacterial communities  and soil  bacterial communities (GS and RS). Among the endophytes from different plant tissues, there were 983 OTUs from roots, 561 OTUs from stems, and 353 OTUs from tubers. Endophytic bacteria from these three tissues types shared 214 OTUs, accounting for 21% of RE, 38% of SE, and 60% of TE (P < 0.05). Meanwhile, in samples with high PCS severity (H), Chao1, ACE, and Obs indices of roots (REH) were only significantly higher than those of tubers (TEH) but not stems (SEH). PCS significantly reduced Ace and Obs indexes of RE without significantly reducing the indexes of SE and TE. There were no significant differences between SE and TE.A total of 2,455 OTUs were obtained from plant endophytic (0% of TE . From OT0% of TE . In sampP < 0.005). When we focused on the effect of PCS, only GS had significant differences between H and L groups, while the endophytic communities from the three types of plant tissues had no significant differences. TE had no significant differences  between H and L groups but showed more marginally significant differences than H and L groups in RE  and SE  communities. The difference between TE and RE of the L group  was smaller than that of the H group , indicating that PCS increases the difference between the tuber and root endophytic communities.To reveal the bacterial community distribution, sPLSDA and ANOSIM were performed . The disWith FEAST, we tracked the endophyte sources of roots, tubers, and stems . The maiProteobacteria, Bacteroidetes, Actinomycetes, and Firmicutes. But we saw differences in the relative abundance of taxa among different plant tissues types and soil samples. For example, at the genus level, Sphingomonas and Pseudomonas were dominant bacteria in SE, Enterobacter and Rhizobium were abundant in TE, Pseudomonas and Enterobacter were abundant in RE and RS, and Rhizobium and Pseudomonas were abundant in GS. Among the five sample types, we identified 769 genera, among which 517 were identified in plant samples and 753 in soil samples, underscoring the richness of microorganisms in soil vs. endophytes on plants.To further characterize the endophyte community composition among different tissues types, the top 10 endophytes at the phylum and genus levels were shown . In genet-test to identify endophytes with significant differences between different plant tissue types. The relative abundance of 35 endophytes was significantly different in different tissues at the genus level  and S. turgidiscabies (YD1-11) from the same sample. We found that OTU62 was 100% identical to the sequence of strain YD1-11, while OTU430 and OTU2835 were not 100% identical with the any pathogenic strains (Streptomyces.The genus  strains . This ilChryseobacterium (four OTUs), Rhizobium (three OTUs), Pseudomonas (three OTUs), Bacillus (two OTUs), Stenotrophomonas (two OTUs), Sphingobacterium (two OTUs), and Streptomyces (two OTUs). Four OTUs, OTU430 (Streptomyces), OTU41 (Chitinophaga), OTU33 (Sphingobacterium), and OTU40 (Cellvibrio), were significantly correlated with OTU62 in more than one network. The degree (the number of other OTUs that are significantly correlated with an OTU) of OTU62 was higher than average, and the most of the degrees of OTUs correlated with OTU62 were higher than average, too. Thus, OTU62 may play an important role in endophytic networks, even if it does not show significant enrichment in samples with high PCS severity from any of the tissue types.We conducted a co-occurrence network analysis to evaluate the importance of OTU62 in endophytic networks of three tissue types . In thesEndophytes are considered to be the second line of defense against plant disease , and theP < 0.05), and the community diversity of roots was higher than in stems and tubers , OTU41 (Chitinophaga), OTU33 (Sphingobacterium), and OTU40 (Cellvibrio), were significantly correlated with OTU62 in more than one network. A multitude of studies have reported that strains of Chryseobacterium spp., Chitinophaga spp., Pseudomonas spp., Sphingomonas spp., and Stenotrophomonas spp. tend to be enriched in plants in response to different pathogen/pest attacks and promote plant resistance . Roots are the gateway for the soil bacterial community to enter the plant. About 50% of root endophytes could be traced to the soil community, especially the rhizosphere community, but only about 20% of tuber and stem endophytes could be traced to the soil. PCS significantly reduced Ace and Obs indexes of RE without significantly reducing the indexes of SE and TE. No significant compositional differences to the endophytes were caused by PCS in the non-lesion tissue types, including roots, tubers, or stems, but did affect the relative abundance of several specific endophytes. PRJNA657530.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: NCBI Sequence Read Archive (SRA) under the BioProject number ZG and BZ conceived and supervised the project. ZG, BZ, WS, and GS designed the experiment. WS, GS, ML, BW, RL, YY, and TW performed the material preparation, the data collection, and analysis. WS, GS, and ML wrote the first draft of the manuscript. All authors contributed to the study conception, design, commented on previous versions of the manuscript, read, and approved the final manuscript.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.


A perennial task is to prevent the occurrence and/or recurrence of most frequent or life-threatening cardiac arrhythmias such as atrial fibrillation (AF) and ventricular fibrillation (VF). VF may be lethal in cases without an implantable cardioverter defibrillator or with failure of this device. Incidences of AF, even the asymptomatic ones, jeopardize the patient’s life due to its complication, notably the high risk of embolic stroke. Therefore, there has been a growing interest in subclinical AF screening and searching for novel electrophysiological and molecular markers. Considering the worldwide increase in cases of thyroid dysfunction and diseases, including thyroid carcinoma, we aimed to explore the implication of thyroid hormones in pro-arrhythmic signaling in the pathophysiological setting. The present review provides updated information about the impact of altered thyroid status on both the occurrence and recurrence of cardiac arrhythmias, predominantly AF. Moreover, it emphasizes the importance of both thyroid status monitoring and AF screening in the general population, as well as in patients with thyroid dysfunction and malignancies. Real-world data on early AF identification in relation to thyroid function are scarce. Even though symptomatic AF is rare in patients with thyroid malignancies, who are under thyroid suppressive therapy, clinicians should be aware of potential interaction with asymptomatic AF. It may prevent adverse consequences and improve the quality of life. This issue may be challenging for an updated registry of AF in clinical practice. Thyroid hormones should be considered a biomarker for cardiac arrhythmias screening and their tailored management because of their multifaceted cellular actions. Atrial fibrillation (AF) and ventricular fibrillation (VF) are clinically relevant and potentially life-threatening arrhythmias. Despite some differences, both AF and VF have been assumed to occur due to abnormalities in the electrical activity involved in impulse initiation and impulse propagation ,6,7,8,9.4), which is a relatively inactive prohormone, and lower amounts of the active hormone, triiodothyronine (T3). About 20% of T3 is made by the thyroid gland and the other 80% comes from T4 upon conversion by type 1 iodothyronine deiodinases (IDs), which is widely distributed. Type 2 ID catalyzes the inactivation of FT4 and FT3 while type 3 ID acts like type 1 ID, and is expressed in skeletal and cardiac muscles, thus providing local intracellular production of T3 [The thyroid gland produces thyroxine  produced by the pituitary gland. This forms part of a feedback loop known as the hypothalamic–pituitary–thyroid axis . The thyyroidism .4 and T3 concentrations are within the reference range. The euthyroid status is defined as the condition wherein the TSH level is within the range from 0.45 to 4.49 mlU/L. In the National Health and Nutrition Examination Survey, 2.5% of the population had a serum TSH below the lower limit of the reference range [Both overt and subclinical (latent) hyperthyroidism are endocrine disorders that occur as a result of excessive TH secretion. TSH is a sensitive indicator of thyroid function. Subclinical hyperthyroidism is common in the general population and its frequency is variable, depending on age, sex, and iodine status ,16,17, ace range .2+ handling, and structural remodeling. Disorders of these cellular factors due to a TH imbalance affect the susceptibility of the heart to arrhythmias [TH affect heart metabolism, electrical properties, and function through the interplay of genomic and non-genomic mechanisms of action ,20,21,22hythmias ,15,17.4 is associated with an increased AF incidence [Both overt and subclinical hyperthyroidism in humans increases the risk for cardiac arrhythmias, especially AF ,23,24. Rncidence . The incncidence ,26,27. 4 administration results in similar increases in circulation, as well as the T3 levels, in cardiac tissues [It should be noted that an exogenous T tissues . Evidenc tissues .2+ handling disorders are the sources of the ectopic electrical activity, while re-entry circuits are promoted by the atrial tissue heterogeneity and disorders in the intercellular electrical coupling mediated by connexin (Cx) channels [GJA5 (encoding Cx40), have been identified in AF [Concerning AF, the cardiomyocyte sleeves overlapping the pulmonary veins along with Cachannels ,32,33,34channels , as welled in AF ,37. As aed in AF . TH may Risk factors for AF, such as aging, obstructive sleep apnea, diabetes, hypertension, dyslipidemia, obesity, cancer, renal dysfunction, and thyroid diseases, which are all accompanied by deleterious oxidative stress, may act synergistically to cause AF ,42,43,44AF, as recognized according to an irregular R–R interval and a missing P wave in an ECG, is a highly prevalent arrhythmia promoting heart failure, embolic stroke, and death . Even shManagement of AF includes antiarrhythmic drug therapies that are often ineffective in terminating AF or preventing its recurrences, possibly because these drugs target a single pathophysiological mechanism . CatheteIt should, however, be emphasized that a considerable number of individuals have no AF symptoms ,63, whic4 concentration has been independently associated with AF [4 to T3 is elevated in Graves’ disease mostly due to an increased deiodase-1 activity. Thyroidectomy in Graves’ disease has been reported to abolish the AF load [4 facilitated the induction of AF in animal models as well [TH are one of the factors associated with AF and potential drivers of AF ,22,69. I with AF . Maximum with AF . The inc with AF  and in p with AF ,77, whic with AF . The rat AF load ,81,82,83 as well ,85,86,873, as well as high-normal T4 or lower TSH levels, have been associated with AF recurrence after an arrhythmogenic foci ablation [4 levels influence the success rate of ablation procedures even in the normal range. Likewise, TH replacement therapy exerts an impact on the AF ablation outcome [Moreover, TH affect the outcomes after an invasive treatment of AF. Accordingly, high circulating Tablation ,89,90,91 outcome . Despite2+-handling and facilitated in the presence of an arrhythmogenic structural substrate . All these events are influenced by the modulating factors, such an ischemia or autonomous nervous system (ANS) and hormonal imbalance, including TH. Specific QRS complex patterns, recognized due to hypertrophy, may potentially predict ventricular arrhythmias [Concerning VF, development of this life-threatening arrhythmia is similar to the development of a multifactorial AF ,6,93. VFhythmias . When sthythmias ,54,95,963 have been positively associated with the heart rate, QTc, and negatively associated with the PR interval and QRS duration [Despite recognizing the basic mechanisms that can cause VF, the changes in the cardiac electrical properties remain poorly understood. Electrical disturbances result from the immediate operation of one or other arrhythmogenic mechanisms in different heart conditions. Accordingly, higher levels of total Tduration .Notably, many pathophysiological processes implicated in the development of AF and VF are linked to a mitochondrial dysfunction, which causes an altered calcium homeostasis, an excess of reactive oxygen species formation (oxidative stress), and alterations in the oxygen consumption. Mitochondria are considered to be a metabolic sink and  the targDespite the progress in the treatment of heart diseases and the management of arrhythmias, sudden cardiac death, occurring due to malignant ventricular arrhythmias, remains a major cause of mortality globally . An implIn contrast to the prevalence of AF, the incidence of VF attributed solely to the hyperthyroid status is less common and registered with a frequency similar to that in the euthyroid population . It is lAutoimmunity may alter the myocardial electrical properties promoting idiopathic ventricular arrhythmias ,102. RecOccasionally, acute thyrotoxicosis accompanied by severe hypokalemia can induce a persistent ventricular tachycardia . Unlike Concerning TH, the prevalence of atrial versus ventricular arrhythmias may be explained by the chamber-related differences in the expression of ion and connexin channels, the duration of the effective refractory period, conduction velocity, and local activation time ,110,111,Thyroid dysfunction and diseases are more prevalent in women (about 5–20 times higher than those in men) and their incidence is more pronounced with age . LikewisEV600 mutation positivity and mutations in the telomerase reverse transcriptase promoter may also be prognostic in TCA progression or recurrence [Inflammation and oxidative stress are involved in the pathogenesis of cancer in a manner similar to that in cardiovascular diseases ,124,125 currence ,129. A mcurrence . 4 therapy. Surgery is the primary crucial treatment to remove the tumor and the involved regional lymph nodes, to facilitate an accurate staging of the disease, to minimize the risk of disease recurrence and metastasis, and to facilitate a postoperative treatment with RAI and an accurate long-term surveillance for recurrence [Standard management of the TCA patients includes a total thyroidectomy, a neck lymph node dissection (if indicated), and radioactive iodine (RAI) ablation followed by a TSH suppressive Tcurrence . The acccurrence ,132.Total thyroidectomy followed by RAI has been associated with benefits in high-risk patients, as well as in the decrease in TCA recurrence. Current guidelines recommend an individualized approach for RAI indication  because 4 downregulates the pituitary TSH that promotes growth of the residual malignant thyroid cells [4 is involved in the long-term management of DTCA [4 affects the TH metabolism and results in a stable subclinical hyperthyroid state along with the down-regulation of deiodinases 1–2 and upregulation of deiodinase 3 [Post-surgery suppressive therapy with Tid cells . Thus, l of DTCA . The ben of DTCA . Long-tedinase 3 . Whetherdinase 3 .4, TH analogs possessing a thyroidomimetic activity, such as triiodothyroacetic acid (an acetic acid metabolite of T3) and tetraiodothyroacetic acid (a derivate of T4), reduce the risk of cancer progression, enhance therapeutic effects, and suppress cancer recurrence [Besides exogenous Tcurrence . Howevercurrence ,148,149.4 therapy and the dosage required for the TSH suppression may be reduced over time [The thyroid axis ‘set-points’ are significantly altered after a long-term Tver time . Howeverver time . Neverthver time . In the ver time . The updThere is a link between AF and thyroid nodules, whereby cytokines and growth factors, such as IGF-1, EGF, and FGF, may be involved . MoreoveTCA is considered an independent predictor for AF along with well-known AF risk factors ,154,155.According to a Swedish Nationwide Study, DTCA patients have a higher AF incidence compared to the general population, and females face a slightly higher incidence of cerebrovascular disease . Thus, iIn the context of arrhythmias, an interesting question arises as to whether TCA patients during follow-up exhibit some specific risk factors for cardiac arrhythmias, such as the pro-arrhythmic β1-adrenergic and M2 muscarinic receptor autoantibodies or changes in the myocardial connexins. It has been shown that TSH and TH regulate the expression of connexins in a variety of target tissues, including the thyroid gland . ReducedTH are powerful modulators of electrical properties of the heart ,23,164, KCNA5 (Kv1.5) and KCNB1 (Kv2.1), that contributed to the main repolarizing K+ currents, as well as the HCN2 and HCN4 genes encoding pacemaker If channels [KCNQ1 (KvLQT1) and its regulator KCNE1 (minK) generating the slow component of cardiac delayed rectification, IKs current, was decreased. The up-regulation of Kv1.5 mRNA was greater in the atrium than that in the ventricle.Microarrays analyses showed that the hyperthyroid status increased the expression levels of genes encoding the voltage-gated potassium channel proteins, notably channels . In contIf) current [Electrophysiological studies revealed that the sinus tachycardia was related to the effect of TH on the rate of diastolic depolarization of the pacemaker cells via the increase in the pacemaker HCN2  and rapid delayed rectifier K+ currents , which result in a decreased APD and the increase in the membrane potential during the plateau phase [IK,Ach), which results in a hyperpolarization and shortening of APD [It is noteworthy that the interaction of β1-adrenergic receptor autoantibodies with the target receptor amplifies the L-type Caau phase . Interacg of APD .2+ handling and intracellular free Ca2+ concentration by regulating the transcription of the genes involved in Ca2+ cycling [2+-ATPase (SERCA2a) increase in response to TH [2+ uptake by sarcoplasmic reticulum whereby its Ca2+ overload may facilitate spontaneous Ca2+ leaks. An abnormal Ca2+ cycling and Ca2+ leak through the ryanodine receptor (RyR) in sarcoplasmic reticulum are involved in the occurrence of triggered activity such as EAD or DAD, followed by premature contraction [NIa current, resulting in an increase in intracellular Ca2+ likely via an accelerated reverse mode of Na+/Ca2+ exchanger [2+ and Ca2+ overloads inhibit intercellular communication mediated by connexin channels that may result in a slower or block of conduction promoting re-entrant arrhythmias [Thyroid status affects myocardial Ca cycling . Both gese to TH , along wse to TH . It suggtraction . DAD maytraction . Moreovexchanger . High dihythmias ,174.2+-handling alterations, AF occurrence is promoted by the presence of a structural arrhythmogenic substrate, such as hypertrophy. This is linked with alterations in the expression, phosphorylation, and distribution of the Cx40, Cx43, and Cx45 channels [TH induce a hyperdynamic cardiovascular state accompanied by an elevation of the left atrial pressure and increase in the size of the atria that facilitates a paroxysmal AF . It suggchannels ,162,163.channels . Hyperthchannels ,101,158.channels . Moreover, a cardiac overload of various etiologies (including hyperthyroidism) is associated with an altered myocardial redox state and oxidative stress, which impairs the function of numerous target proteins including the ion and connexin channels, thereby facilitating the development of AF or VF ,6.TH interact with the sympathetic nervous system by augmenting responsiveness to a sympathetic stimulation presumably via modulating the adrenergic receptor function and/or density. Myocardial adrenergic receptor binding sites have been shown to be enhanced by a chronic, as well as an acute treatment with TH . There i2+ due to an altered Ca2+ handling, along with alteration of the expression and function of the HCN, Na+, K+, and Ca2+ channel and an impairment in the connexin channel-mediated cell-to-cell coupling, seem to be crucial factors in the proarrhythmic signaling of TH. Accordingly, it appears that the long-term subclinical hyperthyroid state including a TSH suppressive therapy with L-thyroxine may increase the risk for AF in post-thyroidectomy patients. Though relatively uncommon, perhaps because of a lack of real evidence due to limitations in the screening process [TH regulate multiple nuclear and extra-nuclear processes that operate to maintain the cardiac function. An overt or more frequent subclinical hyperthyroid state due to thyroid diseases disturb this regulation, promoting the development of cardiac arrhythmias, mostly AF. Disorders in the intracellular Ca process , awarene


Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and has high mortality. Biomarkers related to HCC, such as alpha-fetoprotein, and imaging technology, such as ultrasound and computed tomography, have been used to screen and monitor HCC, but HCC is still difficult to diagnose effectively in the early stage due to the low sensitivity of the above mentioned traditional methods. There is an urgent need for noninvasive biomarkers to facilitate the screening and early diagnosis of HCC. With the advancement of next-generation sequencing, genetic biomarkers are becoming the core of cancer diagnosis. Genetic biomarkers such as peripheral blood circulating tumor DNA, microRNAs, long noncoding RNAs, circular RNAs, and exosomes have become the focus of early HCC diagnostics. HCC genetic biomarkers have been implemented in clinical practice. In this review, we describe the available literature on peripheral blood genetic biomarkers in the diagnosis of early HCC. Liver cancer is the fifth most common cancer worldwide and has become the third leading cause of cancer-related deaths, based on reports from the World Health Organization. Hepatocellular carcinoma (HCC) accounts for ~90% of primary liver cancer, and chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection is the main cause of HCC, accounting for 80% of HCC cases worldwide –3. ApproMiRNA is a short single-stranded noncoding endogenous RNA molecule with a length of ~22 nucleotides. It regulates gene expression by targeting messenger RNA (mRNA), leading to degradation or inhibition of translation , 18. MiRA meta-analysis evaluated microRNA-21 as a biomarker for the early diagnosis of HCC. A total of 515 HCC patients and 338 healthy controls (HCs) or patients with chronic hepatitis (CH) were included in six published studies from different laboratories. The pooled sensitivity and specificity of miR-21 for HCC diagnosis were 85.2% (73.3–88.4%) and 79.2% (68.4–87.0%), respectively. The area under the curve-receiver operating characteristic curve (AUC-ROC) was 0.89, and it was concluded that miR-21 is a useful biomarker for the early diagnosis of HCC . HoweverThe miRNA-based targeted therapy of HCC may be a potential strategy for anticancer immunotherapies. Recently, miRNAs such as miR-16, miR-21, miR-155, and miR-34 have been introduced as therapeutic targets in clinical trials for cancer treatment, including multiple solid tumors and hematological malignances, and have improved the overall survival rate of cancer patients , 54. A mMiRNA is not the only nucleic acid used as a biomarker for HCC, but the long noncoding RNA (lncRNA) is also a current research hotspot as a potential biomarker . The lncp18, and enhances epithelial-mesenchymal transition (EMT) to promote the hepatocarcinogenesis and metastasis of HCC by sponging miR-200a-3p and upregulating the expression of Zinc finger E-box-binding homeobox 1. HULC can also promote HCC progression by activating acyl-CoA synthetase long chain family member 1 and inhibiting miR-372 and miR-9 expression  with size in 30–200 nm diameter that are secreted by almost all types of cells. They can be found in most body fluids  and have variable components such as encapsulate lipids and multiple types of proteins as well as nucleic acids including mRNA, miRNA, lncRNAs, and DNA detected inside exosomes, which can reflect the status of host cells . In the tastasis , 136–138Several studies have shown that exosomal mRNA, microRNA, and lncRNA can be used as biomarkers for HCC screening , 139–144The expression levels of lncRNA ENSG00000258332.1 and LINC000635 in serum exosomes of HCC patients were significantly higher than those of patients with CHB. The AUCs of exosomal lncRNA ENSG00000258332.1 and LINC00635 in discriminating HCC from CHB were 0.719 (71.6% sensitivity and 83.4% specificity) and 0.750 (with 76.2% sensitivity and 77.7% specificity), respectively. The AUC for the combination of lncRNAs (ENSG00000258332.1 and LINC00635) and AFP was 0.894 (83.6% sensitivity and 87.7% specificity), and exosomal ENSG00000258332.1 and LINC000635 were positively correlated with lymph node metastasis, TNM stage and overall survival in HCC patients. These data suggest that these lncRNAs could be potential biomarkers for early HCC diagnosis and the prognosis of HCC .Studies have shown that the serum levels of exosomal miR-122 and miR-148a were significantly higher in early HCC than in LC. The AUCs of exosomal miR-122 and miR-148a in discriminating early HCC from LC were 0.816 and 0.891, respectively. The AUC for the combinations of exosomal miR-122, miR-148a, and AFP in discriminating early HCC from LC was 0.947 (87.0% sensitivity and 90.0% specificity). Exosomal miR-122 had the best diagnostic performance, with an AUC of 0.990 (100% sensitivity and 92.0% specificity), discriminating early HCC from CH. However, exosomal miR-122 and miR-148a were not different between the HCC and CH groups; therefore, the combination of these miRNAs and other markers of HCC diagnosis, such as exosomal lncRNAs, could discriminate early HCC from CH . A studyIn 1948, Mandel et al.  first diStudies have shown that peripheral blood cfDNA levels in HCC patients are correlated with HCC, and total blood cfDNA and ctDNA concentrations in HCC patients are significantly higher than in patients with HC , 167. Nop15 and p16, APC, SPINT2, SFRP1, TFP12, GSTP1, and RASSF1A . Since the methylation profiles of HCC tumor DNA and matched plasma ctDNA were highly correlated; they constructed a diagnostic prediction model with these methylation profiles. Their study demonstrated that this diagnostic prediction model showed better diagnostic performance in differentiating HCC from HCs  are considered to harbor the most important mutations in HCC. Studies have shown that regardless of AFP status, the AUC of these six selected mutations genes to diagnose HCC is 0.92 (TP53, CTNNB1, PLCB1, PCLO, ROBO1, BAZ2B, and TTN, and HCC clinical processes were analyzed. Studies have shown that plasma samples have a high degree of consistency with single nucleotide mutations in corresponding tumor tissues. Dynamic changes in single nucleotide mutations were well correlated with the patient's tumor burden, ctDNA mutation profiles accurately assessed the patient's tumor burden with high consistency as compared with imaging results, and ctDNA mutation profiles could discover tumor occurrence early in advance of imaging for an average of 4.6 months and showed superior performance as compared with serum biomarkers AFP and DCP (TP53 p.R249S mutation is more common in HBV- and aflatoxin-associated HCC than in HCC associated with other etiologies in peripheral blood, and the detection of the TP53 p.R249S mutation shows promise but may only be highly specific for HCC in certain regions and races. In China, the TP53 p.R249S mutation detected in HCC cases was higher than that detected in Thailand and Africa (In addition to the changes in methylation status, tumor-specific single nucleotide mutations of various genes have been found in the peripheral blood of HCC patients, including d CTNNB1 –185. Six is 0.92 . In anot and DCP . Therefo and DCP , 189. Std Africa , 191. Etd Africa . For higd Africa , which iAlthough the above described serum genetic markers have better relative sensitivity and specificity, there are still many limitations to be overcome for the diagnosis of early HCC. The main limitation of most studies using nucleic acid molecules as biomarkers of HCC is the limited size of the cohort, and the heterogeneity of HCC should be also considered. A larger patient cohort including patients with different liver disease conditions, such as alcohol-induced HCC, HBV/HCV-associated HCC and HCCs caused by other etiologies, is necessary to achieve a comprehensive analysis, and also the different races can be the basis for the evaluation of the clinical usefulness of the suggested gene biomarkers. Ideal biomarkers not only have sufficient sensitivity and specificity but perhaps more importantly, they must be widely used for monitoring and cost-effectiveness. The cost of separation, processing and amplification of these molecules, as well as quantitative detection and sequencing, might be so high that it would be difficult to apply on a large scale. These small numbers of cohorts can lead to promising results in research, but verification in large cohorts and the establishment of standard cut-off values for screening and diagnostic purposes are still needed. In addition, the establishment of a standardized test is essential for effective screening and diagnosis. Finally, due to the high heterogeneity of HCC, a panel containing multiple genetic and epigenetic alterations as well as protein and metabolite biomarkers could achieve the best diagnostic sensitivity and specificity to distinguish HCC from liver fibrosis or cirrhosis. CancerSEEK analysis detects eight tumor-associated protein biomarkers and mutations in 1933 distinct genomic positions with 98% sensitivity and 99% specificity for HCC detection with an estimated cost of $500 to perform a test .TS, LL, and BS conceived of the presented idea. TS, SW, and BS wrote the article, and BS supervised the manuscript. All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication. All authors read and approved the final manuscript.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.


Patient adherence to guidelines is important for improved outcomes and prognosis. Nevertheless, many patients with type 2 diabetes mellitus (T2DM) do not comply with the recommendations regarding medication, physical activity, diet or self-care. The present cross-sectional study aimed to assess the level of adherence to the dietary recommendations issued by the American Diabetes Association (ADA) among patients with T2DM in Komotini, Greece. A total of 162 adults with T2DM (64.7 ± 10.6 years old), of which 41.4% were men, were recruited from the Sismanoglio Hospital and participated in the study. The level of adherence to individual recommendations issued by the ADA was assessed using yes/no questions. The overall adherence rate to the guidelines was low (41.2%). According to the multivariable analysis, age and medication therapy were identified as contributors to the compliance rate. No differences were noted in the total compliance rate between patients of different religious denominations (Muslims/Christians). Patients on oral antidiabetic agents (OAA) were more adherent compared with those on insulin therapy. A mere 3.7% of the participants had received nutrition education by a registered dietitian, 9.9% were following an individualized diet plan to improve glycemia, and 3.1% had set specific energy goals to reduce body weight. These findings are indicative of the need for the delivery of improved nutrition education. Diabetes mellitus (DM) is a chronic condition affecting more than 451 million adults worldwide , demonstWith regard to self-management, assessment of blood glucose concentrations is performed on average on 0.2–2.2 days each week, whereas foot care is implemented for approximately 2.2–4.3 days/weekly .Following the lifestyle recommendations is another issue of concern among patients with DM . AnxietyPoor compliance is associated with poorer disease prognosis, an increased risk for developing comorbidities and complications , and a hn = 9), they were excluded from the analysis. Thus, the final sample consisted of 162 patients with T2DM.A total of 171 patients with a diagnosis of DM were recruited from the “Sismanoglio” General Hospital in Komotini, Northern Greece, during the year 2019 (25 June to 16 December). All out-patients visiting the clinic at the time of the study, fulfilling the inclusion criteria, were recruited. Pre-existing nutrition education was not a pre-requisite for participation. Given that the number of recruited patients with type 1 DM was low (Inclusion criteria were as follows: (1) adult patients, (2) diagnosis of T2DM, (3) ability to understand and articulate effortlessly in the Greek language, (4) willingness to participate, and (5) provision of informed consent. Exclusion criteria involved (1) patients not capable of consenting, (2) those unable to understand and communicate in Greek and (3) underage patients. The characteristics of the sample are provided in The study’s permission was granted by the “Sismanoglio” General Hospital Directorate , and all patients provided informed consent before participation. Data were handled according to the Helsinki Declaration for research on humans.According to the World Health Organization (WHO) ,19, adheThe percentage (%) of adherence to the ADA’s recommendations was calculated based on the sum of adherence  according to the 17 questions for patients on insulin therapy and based on the sum adherence of 15 questions for patients on therapy with oral antidiabetic agents (OAA) only.For the purpose of the study, an experienced registered nutritionist dietitian (RDN) (S.K.) was present at the outpatient clinic for DM every day, throughout the study’s recruitment period. Participating patients were interviewed by the RDN on a one-on-one basis, and their reported adherence to the ADA recommendations was recorded on the questionnaire, on paper form, by the same RDN. No training was provided to the participants prior to participation.For specific questions, participants were assisted with photographs of common food portion sizes and the micronutrient content of selected food groups, by the RDN. For instance, with regard to fiber, adequate intake was presented as daily portions of vegetables, fruit, or grains. Concerning sugar-sweetened beverages (SSBs), commercially available choices were presented to participants in order for them to report their intake. Alcohol portions were also presented in picture format, for participants to better understand and report their consumption. Commercially available sugar substitutes and dietary supplements were also presented, to achieve a more valid reporting.The participants’ body weight and height were measured during morning hours with a digital scale  and a wall-mounted stadiometer . Body mass index (BMI) was calculated for each patient by dividing body weight (kg) by the stature (m) squared. Weight status was identified according to the World Health Organization thresholds for BMI [1c assessment, education level, and medication therapy for DM were recorded for each participant. Given that Komotini has the greatest Muslim population in Greece, the religious denomination of participants was also recorded.Data concerning disease duration, comorbidities, latest HbAn) with their respective percentages in parentheses. Chi-square tests and the Mann–Whitney test were utilized to examine differences among qualitative and quantitative variables, respectively.Normality in distribution was assessed both visually and with the Shapiro–Wilk test. Normally distributed variables are presented as mean ± standard deviation (SD) and non-normally distributed as median with the respective interquartile range (IQR). Descriptive statistics for qualitative data are presented as frequencies (2 (continuous), educational level (primary vs. secondary or tertiary), religious denomination (Christian vs. Muslim), diabetes duration in years (continuous), HbA1c in % (continuous), and therapy type (insulin vs. OAA only). A multivariable linear regression analysis was conducted, which included those independent variables selected by the augmented backward selection procedure as previously described [A univariable linear regression analysis was conducted in order to assess the impact on percentage adherence to the ADA’s recommendations total score (dependent variable) of the following study variables: age in years (continuous), sex , BMI in kg/mescribed . NormaliTotal sample adherence to the ADA dietary recommendations  and adheA mere 3.7% of the sample had received nutrition education by an RDN. The majority of participants reported adhering to the fiber (87.7%) and fish intake (78.4%) recommendations; most reported being aware of how to combat hypoglycemia (97.5%) and of the fact that supplements cannot improve glycemic control (79.6%). Only 9.9% of the patients followed an individualized diet plan to improve glycemic control, and 3.1% had specific energy targets set to reduce their body weight. Among those on insulin therapy, only 1.2% were competent to count their meals’ carbohydrate content, and 3.7% were receiving fixed insulin doses paired with a consistent carbohydrate intake. Recommendations regarding alcohol intake were only adhered to by 3.7% of the sample. The vast majority (87.7%) reported maintaining the pleasure of eating. Only 17.9% and 21.6% of the participants were able to control their blood lipid and blood pressure levels by diet alone, without pharmacotherapy.p ≤ 0.001). In parallel, men were more compliant compared with women  and younger patients compared with the older ones .Patients on OAA therapy only, exhibited increased adherence compared with those receiving insulin  demonstrated a negative association with percentage of ADA adherence score . Religious beliefs and BMI were not associated with the ADA adherence score .In a univariable assessment , patientIn the multivariable model , patient1c, and Muslin denomination reduced compliance, whereas, having attained secondary or tertiary education and being on OAA therapy only, were associated with increased compliance. In the multivariable analysis, only OAA therapy and DM duration were associated with the rate of adherence to the recommendations.Patients with T2DM exhibited low compliance to the ADA dietary recommendations for DM. Among those on OAA therapy only, men and younger patients reported a better adherence rate compared with patients receiving insulin, women and older patients. Based on the univariable analysis, age, BMI, DM duration, HbA1c concentrations. In an RCT comparing adherence to the ADA dietary recommendations against a low-fat vegan diet, an improved overall adherence rate was noted in the latter compared with the former [1c concentrations.In the present sample, only 3.7% of the participants had received nutrition education by an RDN. According to a systematic review and meta-analysis, the delivery of dietary education by RDNs consists of the most effective medium to improve patient outcomes . Previoue former , indicate former  failed te former . Based oe former , changinApart from nutrition education, delivery of medical nutrition therapy (MNT) is important, as it involves the nutrition care process model, setting individual goals and guiding patients towards their attainment, close monitoring and evaluation. In the present sample, 9.9% of the patients were following an individualized diet plan to improve glycemia, whereas 3.1% had set specific energy targets and adhered to an MNT scheme aiming to reduce body weight. Individualized nutrition therapy carries more benefits for the patient than receiving dietary advice alone . This isAmong participants in the present sample, complying with the alcohol recommendations proved to be a difficult task, as only 3.7% of them reported adhering to this goal. In contrast to the findings herein, in Northern California , most paConcerning the use of oral nutrient supplements (ONS) to improve glycemic control, most participants (79.6%) were aware that they are not efficient unless a deficiency is apparent, and subsequently, most patients herein abstained from their use. Nevertheless, 6.8% reported consuming cinnamon supplements, 1.9% received aloe vera ONS, 0.6% were reliant on curcumin to aid glycemic control, and 8% were consuming other supplements, with a special focus on poly-unsaturated fatty acids (PUFA). According to a recent scoping review , for manAn individual’s diet is greatly dependent on an interplay of religious, familial, psychological, financial, and personal factors . AccordiBased on the multivariable model, each yearly increment in the age reduced total adherence score by −0.23%, whereas those on OAA therapy only had an increased dietary compliance score by 8.36%. These findings agree with previous research on patients with T2DM, suggesting that the use of insulin is associated with a decreased adherence to the therapy . With reThe results also indicated a reduced adherence rate among women participants as compared to their male counterparts. Previous research on Greek patients with DM has also revealed a reduced nutritional knowledge and self-management practices among women with T2DM inhabiting the city of Agrinio, in Central Greece ,26. AparConcerning adolescents with T1DM , accordiThe present study indicates that MNT and nutrition education are often neglected areas of DM care in Greece. Most hospitals fail to employ an adequate number of RDNs, and, thus, nutritional education and care are often offered in private practice to those patients who have the financial means to address relevant out-of-pocket expenses. On the other hand, multidisciplinary teams are required for the management of all conditions/diseases, and as far as DM is concerned, systematic reviews and meta-analyses have highlighted the importance of including RDNs in routine DM care for improved patient outcomes .The limitations of this study include the assessment of adherence to the ADA guidelines instead of relevant recommendations from a Greek society/authority. Furthermore, the cross-sectional design does not allow for causal associations. In parallel, using a relatively small sample from one hospital only might not allow for extrapolation of the findings .According to the WHO, among patients with chronic disease, the rate of adherence to lifestyle recommendations is lower as compared to medication ones . A salie


Riceberry rice, a gluten-free grain, contains many nutrient components, including carbohydrates, proteins, certain fatty acids, and micronutrients, as well as bioactive non-nutrient compounds, such as polyphenolic compounds. This study aimed to evaluate the effect of bread made from anthocyanin-rich Riceberry rice on the postprandial glycemic response, glucagon-like peptide-1 (GLP-1), antioxidant status, and subjective ratings of appetite. In the crossover design, 16 healthy participants (six men and 10 women) completed four sessions involving blood collection in the fasting state and at 30, 60, 90, 120, 150, and 180 min after food consumption (50 g of available carbohydrate) in a randomized order: 1) glucose solution, 2) wheat bread (WB), 3) Riceberry rice bread (RRB), and 4) Hom Mali bread (HMB). Consumption of RRB resulted in significantly lower postprandial plasma glucose concentration at 30 and 60 min when compared to HMB. No difference in postprandial glucose concentration between RRB and WB was observed. In addition, postprandial plasma insulin showed a significant decrease in the group which received RRB at 15 and 60 min, as compared to HMB. In comparison with 50 g of glucose, as a reference, the glycemic index (GI) of RRB, WB, and HMB was 69.3 ± 4.4, 77.8 ± 4.6, and 130.6 ± 7.9, respectively. Interestingly, the ferric-reducing ability of plasma (FRAP) level was shown to significantly increase after consumption of RRB. In the meantime, a significant decrease in the postprandial FRAP level was also observed following an intake of WB and HMB. All breads caused increases in the postprandial plasma protein thiol group and had similar effects on hunger, fullness, desire to eat, and satiety ratings. However, consumption of RRB, WB, and HMB did not change plasma GLP-1 and malondialdehyde (MDA) levels when compared to the baseline. The findings suggest that anthocyanin-rich Riceberry rice can be a natural ingredient for gluten-free bread which reduced glycemic response together with improvement of antioxidant status in healthy subjects. Clitoria ternatea beverage attenuated postprandial glucose together with improvement of plasma antioxidant capacity when consumed with sucrose [Polyphenols are the most diverse group of phytochemical compounds distributed in vegetables and fruits. Today, there is an increasing interest in the biological activity of polyphenols because of their beneficial effects on human health. A meta-analysis of epidemiological studies revealed that long-term consumption of a plant-based diet rich in polyphenols has been associated with reduction in the risk of major non-communicable chronic diseases, such as diabetes  and card sucrose . In thisO-glucoside and peonidin-3-O-glucoside [Rice, a staple food in many parts of the world, has been recognized as an excellent source of gluten-free ingredients . Howeverlucoside . Riceberlucoside ,15. Recelucoside . Intereslucoside .Interestingly, Riceberry rice has been used to produce commercial gluten-free rice flour and associated food products. Thiranusornkij et al. (2019) discovered that Riceberry rice flour had higher levels of total polyphenols, anthocyanins, and ferric-reducing activity power (FRAP) than Hom Mali flour. In addition, bread made from Riceberry rice flour has been found to contain a higher proportion of undigested starch, leading to a decreased rate of starch hydrolysis . Despitew/w) of Riceberry rice or Hom Mali rice flour, respectively. All breads were prepared the day before the study. Cross-sections of bread made from wheat flour, Riceberry rice, and Hom Mali rice powder are shown in Riceberry rice and Hom Mali rice were ground and sieved using 150 meshes and made into powder, respectively. The rice powder was stored in a sealed bag at room temperature until use. In addition, the study bread was prepared by KCG Corporation Co. Ltd. using the procedure of Chusak et al. . The RRBn = 20) were enrolled in this trial through advertisements on the university bulletin board and social media. The recruitment of the participants was performed by a researcher at the Faculty of Allied Health Sciences, Chulalongkorn University. Measurements of fasting blood glucose, total cholesterol, triglyceride, and LDL cholesterol were conducted by the Health Sciences Service Unit, Faculty of Allied Health Sciences, Chulalongkorn University. The participants performed a self-administered questionnaire containing the inclusion and exclusion criteria. Inclusion criteria included: age of 20–40 years, BMI 18.5–22.9 kg/m2, a fasting blood glucose concentration of <100 mg/dL, a fasting total cholesterol concentration of <200 mg/dL, a fasting triglyceride concentration of <150 mg/dL, a fasting LDL cholesterol concentration of <130 mg/dL, and being free of serious illness. The exclusion criteria included: known history or presence of gastrointestinal disease, metabolic diseases or type 1 and type 2 diabetes, pregnancy or lactating, smoking, alcohol abuse, known food allergies, intolerance or gastrointestinal problems to wheat, gluten or starchy foods, and taking supplementations or medications known to influence glucose tolerance or gastric emptying. Finally, 16 healthy participants (six men and 10 women) who met these criteria were invited to the laboratory and involved in duplicated oral glucose tolerance tests (OGTTs). The informed consent forms were signed by 16 eligible individuals to participate in the study. This study protocol was approved by the office of Ethics Review Committee for Research Involving Human Research Subjects, Human Science Group, Chulalongkorn University (COA No. 270/2561). The trial was registered with the Thai Clinical Trials Registry (TCTR20190113001) and was conducted between February and July 2019.Healthy young male and female volunteers (The study was carried out using a randomized and crossover design with a 1-week wash-out period. Participants were randomly assigned to white wheat bread (WB), Riceberry rice bread (RRB), or Hom Mali 105 rice bread (HMB) by using a computer to generate random numbers with simple randomization. The study investigator generated the randomization plan, then assigned participants to the study treatments. The foods were tested once, and the glucose reference beverages (50 g glucose) were each tested twice.Participants were asked to consume 250 mL of a drink containing 50 g of glucose and then randomly assigned to consume three types of bread: WB, RRB, and HMB containing 50 g of available carbohydrate , respectAfter overnight fasting, participants arrived at the Faculty of Allied Health Sciences, Chulalongkorn University before 8:00 a.m. on the test day. The intravenous catheter was inserted into an antecubital vein, and a fasting blood sample withdrawn by the registered nurse. Then, the participants were instructed to consume the study food within 10–15 min. The time when the participants stared to ingest the study food was set as 0 min. Further blood samples were obtained before (0 min) and at 15, 30, 60, 90, 120, 150, and 180 min after starting consumption of each study food. Moreover, the subjective ratings of hunger, fullness, desire to eat, and satiety were evaluated by the visual analog scale (VAS) rating using a 100 mm scale ranging from 0  to 100 (“extremely”). The scores were recorded at the time of blood collection . A maximPlasma was obtained from EDTA and NaF tubes before centrifugation at 3000 rpm for 10 min at 4 °C. The plasma samples were taken and kept at −20 °C until analysis. Plasma glucose, insulin, and GLP-1 concentration were measured by using the glucose oxidase method  and Human Insulin and GLP-1 ELISA kit , respectively.3 and then incubated in the dark at room temperature for 30 min. The absorbance was measured at 595 nm. The FRAP level was expressed as FeSO4 equivalents. Moreover, the level of plasma thiol group was measured by an Ellman’s assay [The FRAP level was measured according to a previously published report . The pla’s assay  with a s’s assay . Plasma The sample size for this study was calculated and based on a previous study of postprandial responses that reported a significant reduction in glucose . Therefop < 0.05 considered to be significant. Data were reported as changes from baseline and compared by a repeated measures ANOVA and Duncan’s post-hoc test. The incremental area under the curves (iAUCs) of glucose, insulin, GLP-1, FRAP, MDA, and thiol were calculated by the trapezoidal rule. The iAUCs were statistically performed using one-way ANOVA followed by Duncan’s post-hoc test. The glycemic index (GI) of WB, RRB, and HMB was determined by the mean individual ratios resulting from iAUCs for glucose over 120 min after consumption of each bread and ingestion of the glucose solution multiplied by 100.Data were presented as means ± SEM. All statistical analyses were performed with SPSS software  with The baseline characteristics of the participants are described in Postprandial plasma glucose changes after ingestion of WB, RRB, and HMB are shown in The iAUCs for postprandial plasma glucose of breads are illustrated in Plasma insulin responses after consumption of WB, RRB, and HMB are presented in In comparison to the baseline, consumption of WB, RRB, and HMB had no effect on an increase in postprandial GLP-1. Furthermore, there were no significant differences in postprandial plasma GLP-1 levels and iAUCs of the test breads A,B.When compared to the baseline, consumption of WB and HMB markedly decreased the FRAP level A. IntereConsumption of RRB and HMB did not change plasma thiol concentration from the baseline at 30 min, whereas a decrease in plasma thiol was markedly seen in the group which received WB A. After According to the generation of lipid peroxidation, the levels of postprandial plasma MDA after ingestion of all breads are presented in As demonstrated in Carbohydrates are the most important nutrient to modulate and influence postprandial glucose and insulin secretion in the blood circulation . This isNormally, the glycemic index (GI) is a value indicating the immediate effect of dietary carbohydrate-rich foods on the postprandial glucose level after consumption. The results showed that the order of GI was HMB > WB = RRB. The observed results of RRB represented strong support for the greater reduction of postprandial glucose and insulin. Our findings are in agreement with an in vitro study of Thiranusornkij et al. indicating that HMB had a higher predicted GI than RRB . AlthougDuring carbohydrate metabolism, postprandial hyperglycemia directly promotes oxidative stress by inducing formation of reactive oxygen species, reducing antioxidant defenses, and increasing oxygen radical-induced protein and lipid peroxidation . To thisIn addition to oxidative damage of proteins, increasing production of ROS induces the oxidation of sulfhydryl groups in thiol-containing molecules . The redIn general, MDA is produced from the production of reactive oxygen species or free radicals, indicating the increased rate of lipid peroxidation and glucose autoxidation. The findings revealed that postprandial plasma MDA level did not increase at all following ingestion of all breads when compared to the baseline. Our report is inconsistent with the earlier finding indicating that an intake of sucrose (50 g) or glucose (37.5 g) induced an increase in postprandial MDA in healthy subjects ,35. The In general, GLP-1 has been shown to promote satiety and suppress appetite, and thereby reduce food intake. In the current study, ingestion of the three breads had no effect on increasing postprandial plasma GLP-1. In accordance with our results, consumption of bread containing 50 g of carbohydrates led to a slight increase in postprandial GLP-1 than that of wholemeal bread . GonzaleA postprandial rise in plasma glucose and insulin elicited by the consumption of anthocyanin-rich Riceberry rice bread was less than that following the consumption of HMB. In addition, an intake of RRB led to a higher increase of FRAP level compared with WB and HMB. However, all breads similarly affected the protein thiol group, MDA level, and appetite ratings. The findings suggest that consumption of RRB was more effective on reduction of postprandial glucose than that of HMB, whereas it had a higher ability to improve the plasma antioxidant status than that of WB. Therefore, anthocyanin-rich Riceberry rice can be used as a natural ingredient for gluten-free bread in order to manage postprandial glucose with a concomitant improvement of antioxidant status in healthy participants. Further study is needed to investigate the long-term effects of RRB with different compositions of macronutrients.


The buffalo dairy industry has deep roots in Southern Italy, due to the traditional link with Mozzarella di Bufala, a Protected Designation of Origin cheese with high economic value and market demand. At farm level, strategies aiming to improve milk yield and quality are essential to maximize profitability and dairy chain efficiency. In this study, we analyzed a large data set of Italian Buffaloes  in order to detect the phenotypic sources of variation of milk yield and quality traits, disclose exploitable favorable correlations among milk traits, and provide useful information for dairy buffalo chain stakeholders.The buffalo milk industry has economic and social relevance in Italy, as linked to the manufacture of traditional dairy products. To provide an overview of the current status of buffaloes’ performances on a large scale, almost 1 million milk test-day records from 72,294 buffaloes were available to investigate milk yield, energy corrected milk, fat, protein, and lactose content, and somatic cell score (SCS). Phenotypic correlations between milk traits were calculated and analysis of variance was carried out through a mixed model approach including fixed effect of parity, stage of lactation, sampling time, month of calving, and all their interactions and random effects of buffalo, herd-test-date, and residual. Third-parity buffaloes were the most productive in terms of milk yield, while the lowest solid content was detected in sixth parity buffaloes. A considerable gap between primiparous and multiparous buffaloes was observed for milk yield, especially in early- and mid-lactation. Overall, SCS progressively increased with parity and showed a negative correlation with milk yield in both primiparous (−0.12) and multiparous (−0.14) buffaloes. Results suggested that, at the industrial level, milk of primiparous buffaloes may be preferred for transformation purposes, since it was characterized by greater solid content and lower SCS. Results of this study provide a picture of the Italian population of buffaloes under systematic performance records and might be beneficial to both dairy industry and breeding organizations. Bubalus bubalis) is farmed for dairy purposes [Approximately 3% of the world buffalo population is hosted in the Mediterranean area, where the subspecies water buffalo , in the last three decades, the Italian buffalo dairy industry has constantly increased in terms of national market demand and export of cheeses. Both the high quality of products and the favorable selling price have promoted industrial growth [Despite the economic boost, in the PDO area both farm management and knowledge of factors affecting milk-related performances in this species still might be improved. Furthermore, farmers’ sensitivity to milk quality needs to be enhanced. Data collection and analysis on farms, animals, and milk would provide the basis for such potential improvements. On the milk side, the mid-infrared spectroscopy for a fast and cheap analysis is implemented in the official recording schemes of Italian buffaloes since the early nineties, providing phenotypes mostly for breeding purposes. In this paper, part of such data  has been analyzed in order to characterize Italian dairy buffaloes and detect the main non-genetic sources of variation of milk yield  and quality traits.n = 1,414,449) from 106,388 buffaloes reared in 386 farms of Central and Southern Italy (mostly the Campania region) were available along with information on MY, birth, and calving date, stage of lactation, and parity. Test-day records covered the window from January 2013 to December 2017. For buffaloes milked once a day, estimation of MY at 24 h through specific coefficients was available from AIA. Content (%) of FC, protein (PC), and lactose (LC) were predicted from individual milk spectra using buffalo-specific prediction models implemented in the MilkoScan™ FT6000  of the milk laboratory of AIA in Benevento . The somatic cell count  was determined by Fossomatic™  and the energy corrected milk  standardized at 740 kcal was calculated as [For this work, no ethical approval was needed because no experimental trials with living animals were performed. Data were routinely obtained during the official test-day recording system and were provided by the Italian Breeders Association . Test-day records , while SCC was allowed to range from 1000 to 999,000 cells/mL. Values of SCC were converted into somatic cell score , in order to achieve a Gaussian distribution of data: P) were carried out in SAS software version 9.4 .Days in milk (DIM) ranged from 5 to 400 and the maximum parity was fixed at 6. The within parity editing on age at calving was performed with the purpose of excluding buffaloes out of the designed age ranges: 3.32 ± 1.28, 4.66 ± 1.29, 5.91 ± 1.35, 7.08 ± 1.39, 8.24 ± 1.48, and 9.34 ± 1.52 years for first, second, third, fourth, fifth, and sixth parity, respectively. At least 5 test-day records per lactation and a minimum of 5 buffaloes sampled in each herd-test-date (HTD) were kept, ending up with 980,330 test-day records from 162,776 lactations of 72,294 buffaloes in 341 farms. Test-day records belonged to first- (28.04%), second- (24.70%), third- (18.53%), fourth- (13.33%), fifth- (9.41%), and sixth-parity animals (5.99%) and were equally distributed across the years of sampling, with minimum and maximum frequency in 2013 (16.00%) and 2016 (23.74%), respectively. The coefficient of phenotypic variation (CV) was calculated as the ratio between standard deviation and mean of each trait and Pearson’s correlations ; µ is the intercept; Par is the fixed effect of ith parity order; Lac is the fixed effect of the jth stage of lactation; Month is the fixed effect of kth month of calving; (Par × Lac) is the fixed effect of interaction between parity and lactation stage; (Par × Month) is the fixed effect of interaction between parity and month of calving; (Lac × Month) is the fixed effect of interaction between stage of lactation and month of calving; ID is the random effect of lth buffalo; HTD is the random effect of mth contemporary group (HTD); and e is the random residual term. To model lactation stage, 11 blocks of 30 DIM each were created, except for the first, the second-last and the last stage, whose windows were 5–30, 271–310, and 311–400 DIM, respectively. Part of the buffaloes had repeated observations across different lactations. Considering the strong seasonality of the species, the month of calving (12 levels) was included as a fixed effect. The analysis of variance allowed missing values and was carried out in ASReml software version 4.1 [Average MY was calculated for each year of sampling ; all valThe average SCS was the highest in 2017 and the lowest in 2013 ; in partP among all investigated traits were calculated separately for primiparous and multiparous buffaloes  and between SCS and PC (0.03) are available in literature for the same species [P of MY with FC and PC were calculated in both primiparous and multiparous  in inflamed mammary glands is responsible for the lower LC in the alveolar lumen, i.e., in milk [P of LC with both FC and PC obtained in this study were expected (P between FC and PC was positive and moderate in both primiparous (0.43) and multiparous (0.39), slightly greater than the overall rP (0.31) calculated for buffaloes in Brazil [P (0.36 and 0.48) reported for cattle [P (0.96) was found between MY and ECM  affected variability of buffalo MY and quality traits. p < 0.05; data not shown) revealed MY of buffaloes calving in February and April to be equal to MY of buffaloes calving in March. In addition, least square means of July, August, and September were also statistically identical and were all different from least square means estimated for March, February, and April at p < 0.001. The effect of the calving period on MY and composition has been already investigated in buffaloes reared in Brazil [All fixed effects were highly significant in explaining the variability of studied traits. With regard to the effect of the month of calving, it is worth to highlight that buffalo is a short-day breeder species. This means that the reproductive activity increases when daylight hours decrease or, when light hours increase but dark hours are still prevalent . Therefon Brazil  and no dHowever, in Brazil, MY of buffaloes that calved during autumn and winter (5.59 ± 0.10 kg/d) was different than that of buffaloes that calved during spring and summer (5.29 ± 0.06 kg/d) . ConsideThe interaction between the effect of parity and stage of lactation for all studied traits is depicted in Overall, the lactation curves of FC, PC, and LC were comparable to those reported in studies based on smaller datasets . GeneralNon-genetic sources of variation of MY, ECM, composition traits, and SCS were investigated on a large scale in Italian dairy buffaloes. The effects of month of calving, parity, stage of lactation, and their interactions were significant for all traits. Both MY and ECM exhibited a clear gap between primiparous and multiparous animals, especially in early- and mid-lactation. Overall, buffaloes in third parity were the most productive in terms of MY, while the lowest FC and PC were observed in sixth parity buffaloes, particularly in mid- and late-lactation. Overall, LC tended to decrease during the productive life, while SCS progressively increased. Supporting this, SCS was negatively correlated with LC, MY, and ECM. The correlation between SCS and MY was weak but stronger (more unfavorable) in multiparous than primiparous buffaloes. In general, summer calving buffaloes were characterized by lower MY compared to those that calved in the rest of the year, likely due to managerial milking choices related to the milk market demand. In fact, in this study, the most productive buffaloes calved in March in order to maximize productivity when the period of maximum cheese demand is covered. Finally, findings suggested that milk of primiparous buffaloes might be more suitable for cheesemaking compared to milk of multiparous buffaloes and that major attention should be given to milk SCS at individual level to avoid loss of MY and coagulation ability. However, milk coagulation traits collected on a large scale should be used for validation. This paper aimed to provide a phenotypic characterization of milk performances of the current Italian buffalo population under control and may be a useful base for all dairy buffalo stakeholders to optimally address farming techniques and develop strategies useful at both farm and industrial level.