Appropriate management of anemia in patients with hemodialysis (HD) involves the administration of iron supplementation and erythropoietin-stimulating agents (ESAs), in addition to monitoring the response. This study aimed to evaluate the treatment of anemia in patients with HD and describe the factors associated with it and its effect on health-related quality of life (HRQOL).The study was cross-sectional in design. The patients were included from three dialysis centers in Palestine from June to September 2018. The data collection instrument consisted of two portions; the initial portion contained demographic and clinical information on the patients, while the second consisted of the European Quality of Life 5-Dimension Scale (EQ-5D-5 L) and the visual analog scale EQ (EQ-VAS).The study included 226 patients. Their mean age (± SD) was 57 ± 13.9 years. The mean level of hemoglobin (Hb) (± SD) was 10.63 ± 1.71 g/dl, and 34.1% of the patients had a Hb level of 10-11.5 g/dl. All patients who required iron supplementation received it intravenously with a dose of 100 mg of iron sucrose. Almost 86.7% of the patients received darbepoetin alfa intravenously at 0.45 mcg/kg a week, and 24% had a Hb level > 11.5 g/dl. There were significant associations between the level of Hb and the number of comorbid diseases and the ESA that was received. However, other demographics and clinical factors did not significantly affect Hb levels. Certain variables, such as exercise, were a predictor of a higher quality of life. It should be noted that there is a significant impact of a low Hb value on the EQ-VAS scale.Our study found that more than half of the patients had a Hb level below the recommended goal of Kidney Disease Improving Global Outcomes (KDIGO). Furthermore, a significant association was found between patients’ Hb level and HRQOL. Therefore, the appropriate treatment of anemia in patients with HD should be followed by adherence to the guideline recommendations, which consequently improves the HRQOL of HD patients, in addition to obtaining optimal therapy.The online version contains supplementary material available at 10.1186/s12882-023-03254-7. Hemodialysis (HD) is the backbone and life-sustaining therapy for patients with end-stage renal disease (ESRD) who cannot undergo renal transplantation –3. The rAnemia contributes to mortality and morbidity , adverseThe most inclusive objective for managing patients with ESRD is maintaining a long survival rate and preserving the patient’s life at a satisfactory level . Good anA previous study found that 45% of patients with HD had a mean Hb level between 11.0 and 12.0 g/dl, which was according to the KDOQI recommendation, 30% were between 10.0 and 11.0 g/dl, 10% had a level lower than 10.0 g/dl, and 15% were above 12.0 g/dl. The study also showed that women and patients with a low albumin level had a statistically significant low Hb level . The EurIn Palestine, an observational retrospective study assessed compliance with the management guidelines among HD patients. As a result, 8.9% and 43% of the patients achieved the Hb goals according to the National Kidney Foundation Kidney Disease Outcomes Quality Initiative (NKF-KDOQI) and KDIGO, respectively. Additionally, the study found an insignificant association between anemia control and sociodemographic and clinical factors of patients .Treatment of anemia in ESRD is challenging for physicians and patients , as inapOur study is the first in Palestine to focus on this issue. Therefore, this research provides a framework and information that can be a reference to improve anemia management. Additionally, determining factors associated with a low Hb level among HD and evaluating the treatment strategy can help improve the treatment approach of these patients.This study was cross-sectional in design, achieved using a convenient and clustered sampling technique. Participants were recruited from three dialysis centers in the north, center and south West Bank, Palestine: An-Najah National University Hospital in Nablus, Ramallah’s Sons Ward of Palestine Medical Complex in Ramallah, and Al Hussein Government Hospital in Beit Jala.In 2016, the West Bank, under the Ministry of Health, operated a total of 11 hemodialysis units, which were equipped with 148 hemodialysis machines. Furthermore, An-Najah National University Hospital in Nablus had one unit with 35 hemodialysis machines. As a result, the West Bank was able to offer consistent dialysis services to 1,119 patients [http://www.raosoft.com/samplesize.html), with a response distribution of 50%, a 95% confidence interval, and a margin of error of 5%. The calculated minimum effective sample size was 215. Furthermore, the estimated sample size was increased by 5-10% to improve the reliability of the present study and decrease false results.According to the annual health report for Palestine 2016, the number of patients who underwent dialysis in the three centers was 484 . This nuAn-Najah National University Hospital received 47.8% (108 patients) of the sample, Ramallah’s Sons Ward received 32.7% (74 patients) and Al Hussein Government Hospital received 19.5% (44 patients). Furthermore, a pilot study of 10 patients was conducted before starting the actual study, which was not included in the analysis.Data were collected from the three dialyzes from June to September 2018. The questionnaire was completed throughout the working time of each center by interviewing the enrolled patients for almost 10 min and accessing their profiles.The questionnaire consisted of two parts. The first part contained the patient’s sociodemographic and clinical information divided into the following:  SociodemThe second part consisted of the European Quality of Life 5-Dimensions scale (EQ-5D). The Euro Quality of Life (QOL) Group developed the EQ-5D instrument, consisting of the descriptive system and the visual analog scale (EQ-VAS). The descriptive system contains five components . Furthermore, EQ-VAS records the respondent’s self-rated health on a 20 cm vertical visual analog scale with endpoints labeled ‘the best health you can imagine’ and ‘the worst health you can imagine’. EQ-VAS records how good or bad a patient’s health status is today by marking ‘X’ on the scale and then writing the number in the box below. The Arabic form of the EQ-5D has been used in several populations –27 and, ESRD patients who were on maintenance HD, 18 years and older, and accepted participation were included. However, we excluded patients who were unable to understand the question, such as psychiatric patients, cancer patients, or actively receiving chemotherapy.Approvals from the Institutional Review Board (IRB), the Palestinian Ministry of Health (PMOH), and An-Najah National University Hospital were obtained before the initiation of the current investigation. Furthermore, verbal consent was obtained from patients individually, and only patients who agreed to participate were included.The data were analyzed using version 21 of the Statistical Package for Social Sciences (IBM-SPSS) program. The analysis involved presenting the data using the mean (± standard deviation), median (lower-upper quartile), frequencies, and percentages. To assess normality, the Kolmogorov‒Smirnov test was employed. Correlations between the categories of Hg level and other factors with the quality of life scales were examined using the Kruskal‒Wallis and Mann‒Whitney U tests. The EQ-5D-5 L crosswalk index value calculator, based on values from the general population of the United Kingdom, was used to determine the EQ-5D scores. Differences in categorical variables were evaluated using the chi-square or Fisher exact test. Multiple linear regression analysis was conducted to identify the determinants of poor HRQOL. The significance level was set at a p-value of less than 0.05.n = 121, 53.5%) were male. The highest proportion  of patients lived in rural areas, followed by urban areas . The marital status reported by the patients was as follows: 78.8% were married, and 21.2% were single/divorced/widowed. Regarding the level of education, the majority  of the patients had secondary education, followed by  a primary education. A small proportion  of the patients reported having a current job, while 124 (54.9%) were previously employed. The majority  of the patients were nonsmokers, and only 7.5% exercised  was 57 ± 13.9 years, ranging from 21 to 87 years. More than half of the patients  was 4.0 (3.0-5.1). Most patients suffered from hypertension (87.6%), followed by diabetes mellitus (DM) (56.2%) and ischemic heart disease (42.5%)  (years) of renal failure and HD of the study participants was 4.0 (2.0–6.0) and 2.5 (1.0–5.0), respectively. Furthermore, the median number (interquartile range) of HD sessions per week was 3.0 (3.0–3.0), and the median number of hours (interquartile range) of HD sessions was 3.5 (3.5-4.0).n = 71, 31.4%) was the most popular herb used, followed by mixed herbs   used herbal remedies. Arabic gum , calcium carbonate , alfacalcidol  and iron sucrose  were the most commonly used medications  of 6.0 (5.0–8.0). Darbepoetin alfa (The mean Hb level (± SD) was 10.63 ± 1.71 g/dl, ranging from 6.10 to 17.31 g/dl. According to the definition of anemia by KDIGO, 109 (90.1%) male patients were anemic with Hb ˂ 13 g/dl, and 83 (79%) female patients were anemic with Hb ˂ 12 g/dl. Indeed, 84 (37.16%) patients had Hb values < 10 g/dl, 134 (59.3%) patients had Hb values ˂ 11 g/dl, 92 (40.7%) had Hb values > 11 g/dl, and 77 (34.1%) had Hb values 10-11.5 g/dl.Table The results showed that 196 (86.7%) patients used darbepoetin alfa; the mean dose administered (± SD) was 42.1 ± 16.2 mcg. Among 196 patients, 122 (62.2%) received 30 mcg, 69 (35.2%) received 60 mcg, and five (2.6%) received 90 mcg.n = 6) had a Hb level > 13 g/dl. However, of the 84 patients with Hb levels < 10 g/dl, 2.4% (n = 2) did not start ESA therapy. Among the 196 patients who received darbepoetin alfa, 24% (n = 47) had a Hb level > 11.5 g/dl.Of 30 patients (13.3%) who did not receive ESAs, 20% (p = 0.045) and receiving darbepoetin alfa (p < 0.001)  of the EQ-5D-5 L index and the EQ-VAS score were 0.69 0.49–0.81) and 60 (50–75), respectively. Factors such as marital status, having a higher level of education, and a lower number of comorbidities were predictors of Eq. 5D5L. Participants who exercised had higher median HRQOL scores (0.82 (0.79-1.00) for Eq. 5D5L and 70.00 (70.00-89.50) for EQ-VAS) than those who did not exercise (0.66 (0.44–0.79) for Eq. 5D5L and 60.00 (50.00–70.00) for EQ-VAS). However, other variables, such as duration and frequency of hemodialysis, were not significantly associated with the two HRQOL measurement scales Tables  and 4.9–0.81 anp = 0.039) and EQ-VAS (p = 0.004). According to the multivariate analysis, a low level of hemoglobin was the predictor of poor quality of life on the EQ-VAS scale only (Table The median (interquartile range) of the EQ-5D-5 L index for patients with a Hb level < 11 was 0.65 (0.42–0.79), while it was 0.73 (0.51–0.82) for those who had a Hb level ≥ 11. Furthermore, the median EQ-VAS score for patients with Hb levels < 11 was 50.0 (43.8–70.0) compared to 60.0 (50.0-78.8) for patients with Hb levels ≥ 11. The hemoglobin level was found to significantly affect both scales of quality of life: EQ-5D-5 L  was lower than that of a Saudi Arabia study (11.16 g/dl)  and sligHowever, the current study reported that the mean dose (SD) of darbepoetin alfa administered to HD patients was 42.1 ± 16.2 mcg, with the majority receiving the 30 mcg dose. Furthermore, a high percentage of patients (76%) who received ESA had a Hb level ≤ 11.5 g/dl. This could be due to hyporesponsiveness to ESA. Several factors influence the hyporesponsiveness of ESA, including noncompliance, iron deficiency, hyperparathyroidism, inflammation, and bone marrow disorder .According to the NKF-KDOQI and KDIGO recommendations on achieving serum ferritin > 200 ng/ml and > 500 ng/ml, 57.6% and 46.8% of our sample had achieved these goals, respectively. Importantly, serum ferritin and TSAT were available for 72.1% and 81.9% of the patients, respectively. The mean serum ferritin concentration (SD) was 447.49 ± 464.45 ng/ml, while the mean TSAT was 32.33 ± 17.16%. These values were lower than those previously found, 693 ± 420.5 ng/ml for serum ferritin and 38 ± 19.7 for TSAT [Iron administration in patients with HD is frequent. Therefore, iron status is mandatory, as it contributes to anemia and hyporesponsiveness to ESA therapy. However, notable differences were found in the percentage of patients who received iron supplementation and ESA between our results and the findings of the previous Palestinian study, which were 69.9% and 5.1%, respectively . The fluOur results showed an insignificant association between Hb level and any documented demographic characteristics. A previous study from Saudi Arabia found that Hb levels were not significantly associated with age. However, the females had a statistically significant low Hb level . FurtherThe present data revealed that the medians (interquartile ranges) of the EQ-5D-5 L index and the EQ-VAS score were 0.69 (0.49–0.81) and 60 (50–75), respectively. The results of the EQ-5D-5 L index and the EQ-VAS score of a study conducted in Palestine were 0.41 (0.06–0.77) and 50 (50–70), respectively . This diThis study found a significant association between quality of life and hemoglobin targets. A similar finding was found in a study carried out in Iran , which sThis research is the first in Palestine to study the treatment of anemia in patients with HD and determine its association with the HRQOL, demographics, and clinical characteristics of patients. Furthermore, the data were collected through face-to-face interviews and patient medical records, providing reliable and comprehensive data. However, our study had some limitations. First, it was a cross-sectional study, making the reason and the outcome unclear and the resulting associations difficult to interpret. For instance, we cannot determine whether anemia causes poor quality of life or if a low quality of life can worsen anemia. Second, the patients were chosen using a convenience sampling technique, which can disturb the generalizability of the study findings. Third, although face-to-face interviews provide accurate screening, offer the capture of verbal and nonverbal questions that show the level of discomfort with the question and capture the behavior and emotions, bias could be announced. Last, our study did not determine some clinical variables, such as parathyroid hormone level, infection, inflammation, and certain blood factors that may affect Hb level or ESA hyporesponsiveness. This means that multiple factors are associated with the Hb level, and we recommend studying all clinical and sociodemographic variables that have significant correlations with the Hb level; therefore, we could increase the validity of the findings.Our study found that more than half of the patients had a Hb level below the recommended KDIGO target. Approximately half of the patients received iron replacement supplementation, and most of them received ESA therapy. Although they received ESA, many patients had a high Hb level below the target. Consequently, we recommend choosing the most desirable, cost-effective, and optimal therapy and regularly following up with an evaluation of variables that are associated with an inadequate response to ESA. In addition, the study found a significant association between patients’ Hb levels and HRQOL. Therefore, the appropriate management of anemia in HD patients should be followed by adherence to guideline recommendations, consequently improving quality of life.Below is the link to the electronic supplementary material.Supplementary Material 1


Despite a century of research, our understanding of cement dissolution and precipitation processes at early ages is very limited. This is due to the lack of methods that can image these processes with enough spatial resolution, contrast and field of view. Here, we adapt near-field ptychographic nanotomography to in situ visualise the hydration of commercial Portland cement in a record-thick capillary. At 19 h, porous C-S-H gel shell, thickness of 500 nm, covers every alite grain enclosing a water gap. The spatial dissolution rate of small alite grains in the acceleration period, ∼100 nm/h, is approximately four times faster than that of large alite grains in the deceleration stage, ∼25 nm/h. Etch-pit development has also been mapped out. This work is complemented by laboratory and synchrotron microtomographies, allowing to measure the particle size distributions with time. 4D nanoimaging will allow mechanistically study dissolution-precipitation processes including the roles of accelerators and superplasticizers. In this work, the authors use near-field ptychographic nanotomography to visualize cement hydration in situ. They report hydration features with submicrometer detail including calcium silicate dissolution rates, etch-pit growth rates and water-to-air porosity evolution. Therefore, there are many attempts to decrease the cement CO2 footprint2. In order to rationally design approaches to decrease the embodied carbon content of binders, maintaining the performances, cement hydration understanding is key. Unfortunately, there are many unanswered questions3 regarding the complex dissolution and precipitation processes that lead the setting and early hardening of cements4.Portland concrete is the world’s largest fabricated commodity, ∼20 billion tonnes/yr. The enormous production of Portland cement (PC), ∼4 billion tonnes/yr, results in ∼2.7 billion tonnes/yr of CO3, see Fig. 5 and not the protective membrane theory. Conversely, there is no agreement in the mechanism(s) to explain the transition from acceleration to deceleration, when there is a degree of hydration of just 10-20% and plenty of space for the hydrates to grow. The most advanced theories, recently discussed6, are based on heterogeneous nucleation and growth within confined regions taking into account the initial particle size distributions7, for instance, see the reaction zone hypothesis8. There are alternatives9 like the needle model6 where alite, the most abundant component of PC, hydrates11 to yield nonstoichiometric calcium-silicate-hydrate (C-S-H) gel12, which nucleates and grows as needles. Neither the dissolution of small grains, nor water diffusion, nor etch‐pits coalescence, nor C-S-H gel impingement −alone− can currently explain the transition from the acceleration to the deceleration periods3. The factors affecting the C-S-H gel growth in these two periods, III and IV, are not known. Moreover, the role of etch-pits13 needs to be better understood as well as the consequences of the spatial gap which opens between the dissolving (inward) alite grains and the growing (mainly outward) C-S-H gel15. Finally, the density evolution of the C-S-H gel shells is also unknown.The hydration of PC can be divided into five periods16 and synchrotron17 powder diffraction allow following the phase development with time. These studies yield volume-averaged information which misses any spatial feature like particle size dependence. On the other hand, electron microscopy (EM) techniques yield very valuable information, with high spatial resolution, but they only give snapshots, as the experimental conditions are not compatible with the hydration in relevant conditions. In situ tomography19 can contribute to filling this gap. In cements, modelling20 and microstructural characterization methods21 acknowledge the growing importance of X-ray microtomography (μCT) in their different modalities22. Moreover, μCT is being widely used23 and in particular to follow in situ 4D (3D + time) some specific features of cement hydration33. In the last three years, important advances have been reported including: i) the automated correction for the movement of suspended particles at very early ages34 which allowed to follow in situ PC hydration after water mixing35; ii) to follow the fast dissolution of plaster and the precipitation of gypsum36; iii) the simultaneous use of neutron and laboratory X-ray tomographies for in situ studying the microstructural changes of PC mortars on moderate heating37; and iv) the measurement of alite particle dissolution using fast synchrotron nano X-ray computed tomography39. However, none of these 4D imaging works combine the stringent four requirements needed for carrying out relevant contributions to the understanding of the mechanism(s) of Portland cement hydration at early ages: (i) water to cement mass ratio (w/c) close to 0.50, (ii) submicrometer spatial resolution, (iii) good contrast to be able to identify the different evolving components (more than eight), and (iv) relatively large scanned volume to allow hydration to progress with appropriate particle sampling, the particle sizes of commercial PCs have Dv,50∈10-20 μm. In particular, hard X-ray synchrotron microtomography has not the required submicrometer spatial resolution neither sufficient component contrast40, hard X-ray synchrotron nanotomography has not the required contrast between the components to be able to classify the hydrates39 and soft X-ray synchrotron nanotomography has the contrast but it requires very large w/c ratios and very small fields of view which does not allow the hydrates to growth in relevant condition (i.e. confined space with low water-cement ratios)32.On the one hand, in situ laboratory41, which merges scanning X-ray microscopy and coherent diffraction imaging44, met the first three requirements. Hence, it was applied to several binders within capillaries of ∼40 μm of diameter46 using a photon energy of 6.2 keV. The second study46 provided valuable information about C-S-H gel hydrated for 5 months: an average stoichiometry of (CaO)1.80(SiO2)(H2O)3.96 with a mass density of 2.11 gcm−3 and an electron density of 0.64 e-Å−3. Moreover, it allowed quantifying a 6.4 vol% of a second amorphous component, iron-siliceous hydrogarnet, with 2.52 gcm−3 and 0.76 e-Å−3. Here, we have used PXCT in near-field configuration47 to acquire data in a record-thick capillary of ∼160 μm, employing a higher photon energy, 8.93 keV. This configuration, and the iterative algorithms that allowed the reconstructions44, now meet simultaneously the four stringent requirements with current data collections of 3-4 h allowing 4D measurements of spatially resolved data during the first four days of cement hydration. On the one hand, the relatively slow overall acquisition time is the main limitation of this work, but this is expected to improve, see last section. On the other hand, the excellent spatial resolution and contrast of ptychographic nanotomography gave quantitative values of relevant parameters in the dissolution-precipitation processes like alite spatial dissolution rates and etch-pit growth rate. These values can help to test the above described models.So far, ptychographic X-ray computed tomography (PXCT)2g1−, respectively. PC-52.5, with finer particles Dv,50∼12 μm, was used for the PXCT study in order to have more hydrating particles in the analysed volume. It is noted that, in a very recent nanotomographic study7, researchers milled alite very extensively, all particles<10 μm, in order to fit them within a small field of view (FOV) of ∼50 μm. PC-52.5 was used for the PXCT and laboratory μCT imaging studies and the additional laboratory characterization. PC-42.5, with slightly larger average particle size, was used for the synchrotron μCT imaging study.Two commercial PCs have been used and their laboratory X-ray powder diffraction (LXRPD) patterns were analysed by the Rietveld method, see Fig. 48. As expected, PC-52.5 releases more heat than PC-42.5, mainly because of its finesse. The DoH at the maxima of the heat flow peaks were 19% and 10% for PC-52.5 and PC-42.5, respectively. These values are well-known50 but they cannot be explained by current models.The calorimetric study, see Fig. Figure 46 and detailed in a subsection of the S.I. The w/c ratio of the scanned volume in the PXCT study was 0.41.Two additional in situ X-ray imaging studies were carried out to place the results of the PXCT nanoimaging in context. Emphasis is placed on the accuracy of the results that can only be estimated by comparison. The FOVs were cylinders. Figure 51. Figure Figure 52. The procedures are thoroughly detailed in 38. The spatial resolutions, determined by this approach, were 250(25) nm, 264(25) nm, 272(34) nm, 748(19) nm and 2.21(17) μm, for PXCT-19h, PXCT-47h, PXCT-93h, Syn-μCT and Lab-μCT datasets, respectively. As examples of this procedure, Figs. 53 has also been employed to estimate spatial resolution. Figs. The spatial resolution was characterised by two approaches as recently reported46, see Table -Å−3, for the 19, 47 and 93 h datasets, respectively. The spatial resolution from FSC was 470 and 500 nm, for the 47 and 93 h data, respectively. Hence, radiation damage cannot be discarded but it is small, if any. Thirdly, seven sets of components within the tomograms were identified: air, water, AFt/C-S-H gel/others, CH, Cc, C3A/C3S/C2S and C4AF, using the electron density and the absorption data in the bivariate plots, see Figs. 2, has a theoretical electron density value of 0.69 e-Å−3. The measured values at 19 and 93 h were 0.62(2) and 0.651(5) e-Å−3, see Table 54. This difference is very likely due to the presence of residual water porosity below the spatial resolution of the measurements, which we refer to partial volume effects. It should be noted that individual C-S-H nanoparticles have sizes close to 5 nm55 much smaller than the spatial resolution of this work, i.e. ∼250 nm. Therefore, the C-S-H regions analysed here very likely contain other components like gel and capillary water porosities and interspersed calcium hydroxide. It should also be noted that the employed stoichiometry for C-S-H gel, i.e. (CaO)1.80(SiO2)(H2O)4.057, is an assumption and slightly smaller Ca/Si ratios, ∼1.70, have also been reported58. On the other hand, ettringite and portlandite particles have larger sizes, usually ranging 1-5 μm3, and they can be imaged in the present work. In any case, partial volume effect  is always taking place in cement pastes as some hydrates may have quite small particle sizes17.In situ near-field PXCT data were taken as detailed in methods. To ensure the relevance of the results, the scanned volumes were assessed. Firstly, the w/c ratio was 0.41(2), as determined from the absorption data-Å−3, respectively; when the phase retrieval is carried out quantitatively54. The paste evolution is displayed in Fig. 59 which is out of in situ PXCT capabilities.The crux of our results is the 4D submicrometer features of cement hydration, see Figs. 2S particles were identified and excluded from this analysis, because they do not have C-S-H shells at 19 h of hydration. C3A particles were also recognised and discarded because of their smaller electron density values, i.e. ∼0.91 e-Å−3, 5% lower than that of alite. From 22 measurements along different surfaces, the dissolution rate between 19 and 47 h was 25(14) nm/h. This value compares well with 36 nm/h from the reaction zone model8 but poorly with 84 nm/h obtained from the same dataset by using the boundary nucleation and growth model60. Moreover, this spatial dissolution rate can also be estimated from the segmentation results presented in the next subsection. Based on the quantitative analysis derived from Machine Learning (ML) segmentation of the PXCT datasets  μm of size. Here, their time-evolution can be followed and it is noted that hollow regions are filled with water at 19 and 47 h but dried at 93 h, see Fig. 62 (at 24 h) reporting a maximum size of 5.0 μm with shells of 500 nm. Hence, it seems to be a 3-4 fold difference between the spatial dissolution rate of small alite grains in the acceleration period and that of large alite grains in the deceleration stage.The analysis of the 12 largest Hadley grains 1.8(SiO2)(H2O)4.0 composition, i.e. including gel pore water, had 0.64 e-Å−3  46. As this result is critical for discarding diffusion as the mechanism for the deceleration period, a larger study was carried out. 20 shells were analysed giving 0.51(4) e-Å−3 and 500(120) nm, for the average electron density and thickness, respectively. Moreover, this study also yields the average width of the gap, 490(140) nm. At 24 h, the thickness of the shells and gaps were reported as ∼500 and ∼300 nm from EM62. Moreover, a gap of 490 nm developed in 15 h, time between the end of the induction period and the first nanoimaging measurement, which means an alite spatial dissolution rate of ∼33 nm/h. Therefore, the dissolution rates of large and small alite grains differ at least three-fold, which should be considered for modelling. To further study the C-S-H shells, the components were segmented by ML as described in methods following the procedure summarized in Fig. Figure nano-features. Figure 30. Figure 4AF intergrown regions, evidencing the importance of alkanolamines at early ages as accelerators64. Moreover, Fig. 3A.Finally, Fig. 65. For cement pastes, higher porosity near the capillary wall has been observed by synchrotron microtomography24. For a water-rich alite paste, wall effect was clearly observed by PXCT where the resulting C-S-H gel had higher water content near the capillary wall66. In order to quantitatively study this feature, the scanned capillary was divided into two volumes, a central cylinder with half of the radius and the outer region up to the glass capillary wall. The mean electron densities were computed, but the voxels with electron density smaller than 0.24 e-Å−3, air porosity, were not included in order to minimise any bias due to the shrinkage/pore drying. The results for the centre volume were 0.614, 0.618 and 0.617 e-Å−3 for the 19, 47 and 94 h datasets, respectively. The corresponding mean electron density values for the outer region were 0.608, 0.610 and 0.601 e-Å−3. The 1% difference between the two regions at 19 h is quite small but not negligible. Hence, the degree of hydration could slightly be a function of the horizontal position of the particles.Figure The amount of anhydrous components, together with the initial values, allow determining the DoH from the three imaging studies which are reported in Table 3 or larger. For this particle tracking statistical study, they were arbitrarily classified into four groups having connected volumes (μm3) of 1.0≥vol1 > 27.0≥vol2 > 216.0≥vol3 > 1000.0≥vol4. A rough estimation of their sizes in μm could be ∛(vol) or 1.0≥size1 > 3.0≥ size2 > 6.0≥size3 > 10.0≥ size4. The groups contained 1117, 204, 61 and 20 particles, respectively. The corresponding volume percentages with respect to the overall anhydrous cement particle volume at this hydration age, were 5.4, 12.9, 21.5 and 60.2%. It is not possible to calculate the DoH of these groups at 19 h because there are no reference values at t0. The degree of reaction, between 47 and 19 h, was 69.4%, 59.3%, 37.2% and 15.9% for the groups classified as vol1, vol2, vol3 and vol4, respectively, and they account for 12.9, 26.4, 27.7 and 33.0% of the total dissolved volume. In other words, the twenty particles of vol4 group has a degree of reaction of 15.9% but it accounts for 33% of the dissolved volume in that period. A similar study can be carried out between 93 and 47 h. The degree of reaction during this period decreased to 14.7%, 11.6%, 6.7% and 3.6% for the corresponding groups. In terms of dissolved volume percentage in this period, the values are 6.8, 16.9, 25.3 and 51.0%, respectively. This simple analysis shows that 51% of the observed reaction during this diffusion-limited period is due to the 20 largest particles as they account for a very large fraction of the volume. The strong decrease of hydration rate in this time range is in line with a diffusion controlled hydration stage.Importantly, the high resolution and excellent contrast of the PXCT study allowed us to track down the individual hydration evolution of 1407 particles with connected anhydrous volume, at 19 h, of 1 μmFinally, Fig. 2 is due to a lower density of the C-S-H shells (higher water diffusion) or if it is mainly due to higher calcium supersaturation.The hydration of PC, in relevant conditions, has been measured with very high spatial resolution, i.e. 250 nm, and contrast. As expected but not directly measured so far, the hydration of Portland cement at 1 day or earlier is dominated by the small particles, smaller than 3 μm, and the hydration after 3 days is very much dependent of the large alite particles, larger than about 10 μm. The nanoimaging work shows the C-S-H gel shells surrounding every alite grain which does not preclude diffusion. The measured alite spatial dissolution rates, ∼100 nm/h for small grains in the acceleration period and ∼25 nm/h for large particles in the deceleration stage impose constraints on the cement hydration models. The alite etch-pit growth rate between 19 and 47 h has been estimated as ∼40 nm/h, which decreases to ∼7 nm/h in the 47 to 93 h interval. Moreover, etch-pit coalescence, the merging of different branches, has also been visually observed. However, better spatial resolution is required for a thorough etch-pit growth rate quantification. The configuration employed here already allows studying the roles of admixtures  by measuring C-S-H shell density, C-S-H gel spatial distribution and alite spatial dissolution and etch-pit growth rates. For instance, it would be possible to measure if the acceleration produced by CaCl67. Another approach could be to use machine learning/deep learning for denoising of datasets collected with much less X-ray dose68. On the other hand, gamma irradiation of Portland pastes, mortars and concretes is known to produce water radiolysis finally leading to H2 microbubbles and calcium peroxide, CaO2.8H2O69. Therefore, the signatures of these species should be monitored for studies with high X-ray doses. Higher spatial resolution and shorter acquisition times will allow to thoroughly study the acceleration stage which will be beneficial to rationally design new accelerator admixtures with the final aim of developing low carbon cements with competitive mechanical strengths and durability performances. Moreover, dissolution-precipitation processes with moderate reaction rates take place also in several other fields like geochemistry or biomaterials, which could benefit from the reported investigation.The current spatial resolution of in situ near-field PXCT, i.e. ∼250 nm, could be improved by increasing the number of projections, without larger acquisition times. Moreover, so far the time resolution is modest, i.e. 3-4 h for a complete tomogram, but this is expected to improve in fourth-generation synchrotron sources with tailored beamlines for ptychography. Time resolutions of ∼1 h will open the way to accurately study the processes in the acceleration period. However, in these cases, radiation damage could be an issue if the total dose is not kept low. Higher time resolution does not necessarily imply higher doses and therefore possibly larger radiation damage. For instance, sparsity techniques could be coupled to PXCT in order to decrease the overall acquisition time for the whole series by as much as 90%, as recently reported70.Two different types of commercial cement were used: a CEM I 52.5 R (PC-52.5) and a CEM I 42.5 R (PC-42.5) which conform to EN 197–1. The full characterization of these anhydrous materials is performed with X-ray Fluorescence (XRF) and Laboratory X-ray Powder Diffraction (LXRPD) using the Rietveld Method, see Fig. 2 adsorption isotherms in ASAP 2420 equipment . The air permeabilities were measured with the Blaine fineness apparatus (Controls) according to EN 196–6. The density of the samples was measured with a helium Pycnometer . The resulting data are reported in Table The textural characterization was carried out in dry conditions. Particle size distribution (PSD) data were measured by laser diffraction in a MasterSizer 3000 equipment . Specific surface areas were determined by NThe pastes were prepared using the PC-52.5 and PC-42.5 and with w/c mass ratios of 0.40 and 0.50, respectively. For PC-52.5, 0.43 wt% (by weight of cement) of PCE was used. Water and superplasticizer were mixed by magnetic stirring for 1 min. Then, the cement was mixed with the water/suspension and shaken, for 1 min manually and 1 min with a laboratory vortex mixer. Finally, the pastes were introduced into the glass ampoules. The calorimetric data were taken in an eight-channel Thermal Activity Monitor (TAM) instrument. Data were collected for up to 7 days at 25 °C and at 20 °C and for PC-52.5 and PC-42.5, respectively. The first 45 min were required for the thermal stabilization of the system. The employed conditions mimic the ones used for the imaging studies.1 radiation (λ = 0.7093 Å). This diffractometer is located at SCAI, University of Malaga. The incident beam was formed by a primary monochromator with a focusing mirror and a 2 mm anti-scatter slit. Moreover, 2.5° Soller slits were used for the incident and transmitted beams. An EIGER detector  was used which is optimised for Mo anodes. This was used with an aperture of 4 × 21 degrees, working in VDO mode. Data collection was performed from 3 to 35° (2θ) for 2 h and 10 min. Rietveld quantitative phase analysis was performed with GSAS software.The same paste prepared for the calorimetry measurement (PC-52.5-w/c = 0.40) was used to fill the capillaries of ~1 mm in diameter. LXRPD measurements were collected on a D8 ADVANCE diffractometer (Bruker AXS) using strictly monochromatic Mo-Kα6 source filament and employing a 0.25 mm Al foil to minimise the beam hardening effect. This source was operated at 55 kV and 130 µA. The CCD3 detector with a physical pixel size of 17.427 µm was set in a middle position with a source-to-detector distance of 315.449 mm and a source-to-sample distance of 9.051 mm which yielded a voxel size of 1.00 µm (binning 2 × 2). Finally, the projections were acquired every 0.22° over 360° with a total of 1637 projections per tomogram and using an exposure time of 1.9 s. This results in an overall recording time of 3.5 h per dataset. Image reconstruction of the CTs was carried out using Bruker NRecon software (version 2.1.0.1) and by applying Gaussian smoothing and beam hardening correction.Lab-μCT experiments were carried out at ~25 °C for the same capillary used in the LXRPD data collection and scanning the same region with time. Lab-μCT experiment was performed on a SKYSCAN 2214 (Bruker) scanner at SCAI, University of Malaga. Images were obtained using an X-ray tube with a LaBBecause Lab-μCT and LXRPD data were acquired in the same capillary, but not in the same equipment, the description of the timing is important. Lab-μCT data were collected from 17.5 h until 21.0 h, using cement-water mixing time as the reference. This dataset is labelled as 19 h. Then, the capillary was transfer to the diffractometer, in the same room, and the LXRPD data were taken from 21 h 15 min to 23 h 25 min. This dataset is labelled as 22 h. The data collections at 47 h and 93 h followed the same protocol but it is less important at these hydration ages, as the kinetics are slower.51. The resulting voxel size was 0.65 μm. Further experimental details are given in the SI.PC-42.5 with a w/c ratio of 0.50 was used to prepare the paste for this experiment and PCE was not required. The paste was manually mixed for 3.5 min and then introduced in a capillary of ~0.7 mm of diameter. Inline propagation-based phase-contrast microtomographic data were acquired at ID19 beamline of the European Synchrotron (ESRF) in Grenoble, France. The measurements were performed at 21.5 °C, temperature of the experimental hutch, using a photon energy of 19 keV. The distance between the sample and the detector was 15 mm. The total time to record a full tomogram was 6 min with 0.05 s exposure time. 6000 projection angles were acquired over a 360 degree tomographic scan. For reconstructions, Paganin phase retrieval of the projections was performed47 is a variant of X-ray ptychography44 in which the sample is scanned across a coherent divergent illumination and magnified images of the sample are recorded on the detector at each scanning position. Ptychographic phase retrieval algorithms are employed to reconstruct the complex transmission function of the specimen, with both absorption and phase, at each angular projection. We then repeat measurements of many projections at different incident angles of the X-rays onto the sample and combine them using standard tomographic reconstruction methods to obtain 3D maps of the electron density and the absorption coefficient of the specimen. In near-field ptychography, if a sufficient number of projections are recorded, the spatial resolution is limited by the magnified pixel size, which is determined by the pixel size of the detector, the distance between the sample and the detector, the divergence of the illumination and the position of the specimen from the point source of the beam, i.e. the focus. We performed our measurements using a high-stability instrument designed for high-resolution PXCT working in air72 and at the hutch temperature of 25 °C, using a photon energy of 8.93 keV. The thickness of the capillary in the imaged region was 160 µm. Therefore, it was scanned with a FOV of 186 µm in order to have more than 10 μm of air outside the capillary, which is required for quantitative phase imaging. The total time to record a full tomogram was between 3 and 4 h, including dead time during the motion of stages in between acquisitions. The voxel size was 186.64 nm. Further experimental details are given in the SI.The paste employed for the in situ nanoimaging study was PC-52.5 with a nominal w/c ratio of 0.50 and 0.43 wt% of PCE. The suspension was mixed with a mechanical stirrer at 800 rpm for 3.5 min and then introduced in a glass capillary of 200 μm of nominal diameter. Near-field PXCT data were taken at the cSAXS beamline of the Swiss Light Source (SLS) at the Paul Scherrer Institute (PSI), Villigen, Switzerland. Near-field ptychographyA supervised Machine-Learning (ML) image analysis approach was used to segment the different components of imaged samples, using the IPSDK Explorer software . The plot profiles along the scans electron density/grey value slices and the 3D rendering visualization were done by using Dragonfly software  Inc., Montreal, Canada). More information about data analysis can be found in the SI.Supplementary InformationPeer Review FileDescription of Additional Supplementary FilesSupplementary Movie 1Supplementary Movie 2


Transforming growth factor‐β (TGF‐β) type I receptor (TGFβRI) and type II receptor (TGFβRII) are the members of the TGFβ superfamily, which are potent regulators of cell proliferation and differentiation in many organ systems, and they play key roles in multiple aspects of follicle development.TGFβRI and TGFβRII genes, and the association with litter size in Tibetan sheep.We aimed to explore the characterization, expression analysis of TGFβRI and TGFβRII genes in Tibetan sheep and analyzed their genomic structures.In this study, we cloned the complete coding sequences of Ovis aries (100%), followed by Bos mutus (99%). The RT‐qPCR showed that two genes were expressed widely in the different tissues of Tibetan sheep. The TGFβRI expression was the highest in the lung (p < 0.05), followed by the spleen and ovary (p < 0.05). The TGFβRII expression was significantly higher in uterus than that in lung and ovary (p < 0.05). In addition, the χ2 test indicated that all ewes in the population were in Hardy–Weinberg equilibrium, and the population was in medium or low polymorphic information content status. We also found four Single Nucleotide Polymorphism (SNPs), g.9414A > G, g.28881A > G, g.28809T > C, g.10429G > A in sheep TGFβRI gene and g.63940C > T, g.63976C > T, g.64538C > T, g.64504T > A in TGFβRII gene. Three genotypes, except for g.64504T > A, and three haplotypes were identified in each gene. linkage disequilibrium analysis indicated that there was strong linkage disequilibrium in each gene. The association analysis showed that the four SNPs of TGFβRI were associated with litter size (p < 0.05), and g.63940C > T of TGFβRII was confirmed to be associated with litter size (p < 0.05).The results showed that percentages of sequences homology of the two proteins in Tibetan sheep were the most similar to Based on these preliminary results, we can assume that TGFβ receptors (TGFβRI and TGFβRII) may play an important role in sheep reproduction. TGFβR I and TGFβR II genes, and its association with litter size in Tibetan sheep. The TGFβ receptors (TGFβRI and TGFβR II) may play an important role in sheep reproduction.This study was undertaken to explore the characterization, expression analysis of  The health and reproduction records of the animals were kept by the farmers. Their litter size was obtained from reproduction records. All efforts were made to minimize discomfort during the blood collection. Blood samples were collected from the jugular vein under the supervision of qualified veterinarians. Genomic DNA was extracted from blood sample of each sheep using an EasyPure Blood Genomic DNA Kit . Three ewes were selected from purebred herds of the same farm in Qinghai province. The three selected ewe (6 months old) were healthy, similar in weight, and pastured in similar conditions of grassland. After slaughtered, and the tissues from hypothalamus, hypophysis, heart, liver, spleen, lung, kidney, ovary, oviduct, uterus, rumen, duodenum, and longissimus dorsi were collected and immediately frozen in liquid nitrogen, and then stored at −80°C. The RNA of tissues was extracted by TransZol (TransGen Biotech). Total RNA for each tissue was reverse‐transcribed to cDNA by TransScript One‐Step gDNA Removal.2.2TGFβRI and TGFβRII  were used as templates. The primer pairs were designed using the coding regions of the two genes  competent cell and grown in Luria‐Bertani (LB) agar plate with Amp. White colonies were selected (10 colonies for each sample) and cultured in liquid medium for 5 h, and then sequenced by Shanghai Sangon Biological Engineering Company. Alignments of multiple sequences were carried out with BLAST . Open reading frame (ORF) Finder (http://www.ncbi.nlm.nih.gov/projects/gorf/) was used to determine the ORF and predict the amino acid sequence. ProtParam (http://web.expasy.org/protparam/) was used to predict the physical parameters of each protein. The hydrophilicity and hydrophobicity were analyzed using ProtScale . Prediction of the secondary structure of each protein and its variants was analyzed using SOPMA (http://www.compbio.dundee.ac.uk/www‐jpred/). SWISS‐MODEL (http://swissmodel.expasy.org/) was used to predict the protein signal peptide and protein tertiary structure. Amino acid sequences alignments were conducted using DNAStar Lasergene (MegAlign), and a phylogenetic tree was established using the MEGA7 software.The PCR product was purified using agarose gel DNA extraction kit , and cloned into pMD19‐T vector , then transformed into 2.3TGFβRI and TGFβRII gene (GenBank accession No: XM_004004226.4 and XM_012099309.2) . All experiments were performed in triplicate, and GAPDH was used as the reference gene. The 2−∆∆CT method was used to analyze the data  were screened Using the dbSNP database (http://www.ncbi.nlm.nih.gov/snp) and verified by DNA sequencing. Improved multiplex ligation detection reaction (iMLDRTM) was used for genotyping following the instrument operating guidelines. Genotypic, allelic frequencies, and genetic parameters were directly calculated following previous description  program. All statistical analyses were performed using SPSS 23.0. Results with ijny is the phenotypic value, μ is the population mean, Pi is the fixed effect of the ith parity , Gj is the fixed effect of the jth genotype , PGI is the interaction effect of parity and genotype, and ijne is the random residual.where 33.1TGFβRI gene was cloned, which contained a calculated ORF of 1506 bp encoding a protein of 501 amino acid residues. Additionally, sheep TGFβRII contains ORFs of 1416 bp, and they encode proteins of 471 amino acid residues. The molecular weights of TGFβRI and TGFβRII are 55960.70 and 52879.55 Da, respectively, and the theoretical isoelectric points are 7.19 and 5.84, respectively. All of them include 20 types of amino acid composition. The total numbers of negatively charged residues (Asp + Glu) are 56 and 60, respectively, and the total numbers of positively charged residues (Arg + Lys) are 56 and 49, respectively. TGFβRI formula is C2470H3936N688O723S35. The total number of atoms is 7852. The Aliphatic index is 89.92; the grand average of hydropathicity (GRAVY) is −0.097; TGFβRII formula is C2339H3689N637O703S28. The total number of atoms is 7396. The Aliphatic index is 90.45; grand average of hydropathicity (GRAVY) is −0.170. A positive value indicates that the protein is hydrophobic, and a negative value indicates that it is hydrophilic, so all of them are hydrophilic. Subcellular localization of TGFβRI is 55.6% in endoplasmic reticulum; it is 22.2% in Golgi, 11.1% in plasma membrane, 11.1% in extracellular, including cell wall. And TGFβRII is 34.8% in nuclear; it is 26.1% in cytoplasmic, 21.7% in mitochondrial, 4.3% in endoplasmic reticulum, 4.3% in peroxisomal, 4.3% in vesicles of secretory system, 4.3% in vacuolar. The proteins of TGFβRI and TGFβRII have signal peptides. WEBSEQUENCE Number of predicted Transmembrane Helices is 2 and 1. There were potential N‐glycosylation sites at amino acids 41, 148, 268, and 348. The potential values were 0.7000, 0.8358, 0.6267, and 0.4613 in Tibetan sheep TGFβRI. There were potential N‐glycosylation sites at amino acids 70, 94, and 266. The potential values were 0.5869, 0.6930, and 0.6757 in Tibetan sheep TGFβRII. There are 50 and 45 potential phosphate sites in sheep TGFβRI and TGFβRII, respectively. Amino acid sequence alignment and percentage of sequences homology of the two proteins in Ovis aries, Bos taurus, Bos mutus, Homo sapiens, Sus scrofa, Mus musculus, Maylandia zebra, Canis lupus familiaris, Pan troglodytes, Macaca mulatta, and Gallus gallus showed that Tibetan sheep TGFβRI and TGFβRII are most similar to O. aries (100%), then B. mutus (99%), and least similar to C. lupus familiaris (82%), respectively  and TGFβRII (b) of Tibetan sheep, sheep, cattle, yak, house mouse, human, chimpanzee, rhesus monkey, pig, dog, chicken and zebra with that of Tibetan sheepThe structure prediction of sheep TGFβRI protein was performed by online protein analysis system SOPMA. The results showed that the extension chain composed of alpha‐helix, extended strand, beta turn, and random coil accounted for 39.32%, 11.38%, 3.39%, and 67.27%, respectively, and for TGFβRII protein, they were 33.76%, 15.92%, 3.82% and 46.50%, respectively , followed by the spleen, uterus and ovary (p < 0.05), and almost no expression in longissimus dorsi. The TGFβRII expression was the highest in uterus than in other tissues (p < 0.05), followed by lung, ovary, and spleen (p < 0.05). There were no significant differences among oviduct, duodenum, rumen, kidney, pituitary, liver, and heart (p > 0.05). Except for the hypothalamus, the expression of TGFβRII gene in longissimus dorsi was lower than that in the other tissues (p < 0.05).The RT‐qPCR was used to investigate the general tissue distributions of 3.3TGFβRI and TGFβRII genes, respectively. All mutations were synonymous mutations. Except for SNP g.64504T > A, the other SNPs were classified as three genotypes (Table r2) block indicated strong linkage disequilibrium in two genes, respectively  of Tibetan sheep TGFβRI were 0.72, 0.28, 1.40, and 0.24, respectively, and for TGFβRII, 0.76, 0.24, 1.31, and 0.21, respectively. Tibetan sheep were in medium PIC status at g.63940C > T and g.28809T > C sites, and the others have low PIC status  were identified in Tibetan sheep es Table , and thres Table . Linkagey Figure . In addi3.4TGFβRI and TGFβRII SNPs on litter size of the experimental populations were studied. The results showed that the g.9414A > G, g.28881A > G, g.28809T > C, and g.10429G > A of sheep TGFβRI were associated with litter size (p < 0.05). In contrast, the TGFβRII g.63940C>T substitution was associated with litter size (p < 0.05). However, the SNPs, g.63976C > T, g.64538C > T, and g.64538C > T had no association with litter size  (Grant No. 2020‐ZJ‐786); Outstanding Person of Kunlong: Rural Revitalization Program (Grant No. (2020)9).All experiments in this study were performed following the approved guidelines of the Regulation of the Standing Committee of Qinghai People's Congress. All experimental protocols and the collection of samples were approved by the Ethics Committee of Qinghai University under permission No. SL‐2021027.https://publons.com/publon/10.1002/vms3.1013.The peer review history for this article is available at


Deff), and rehydration rate of sweet cherries (SC). The drying duration of microwave‐treated SC was shorter than the untreated sample. The average drying time of fresh SC microwaved for 0, 30, 60, 90, and 120 s were 220, 205, 190, 175, and 150 min, respectively. The Deff values, total phenolics, and antioxidant capacity of microwave‐treated SC were higher than the untreated sample. In this study, the SC Deff as determined by the second Fick law varied from 8.73 × 10−10 to 1.41 × 10−9 m2/s. The experimental data for the dehydration curves were fitted to different thin‐layer equations, and the Midilli equation using the experimental constants best described the drying rate of SC. As the microwave pretreatment time increased from 0 to 120 s, the total phenolics and antioxidant capacity of dried SC increased from 1491.4 μg Gallic acid equivalents (GAE)/g dry to 2272.1 μg GAE/g dry, and 54.47%–62.59% (p < .05). The microwave pretreatment enhanced the rehydration rate of dried SC. The rehydration percent of dried SC microwaved for 0, 30, 60, 90, and 120 s were 127.27%, 136.63%, 136.91%, 137.07%, and 136.72%, respectively.The target of this work was to investigate the influence of microwave pretreatments  on the total phenolics content, antioxidant potential, mass transfer rate, effective moisture diffusivity ( Microwave pretreatment not only increases drying rate and excellent energy efficiency, but also improves the retention of nutrients and total phenolics. Our results showed that the drying rate of sweet cherries during drying can be significantly improved by microwave pretreatment. MicrowaPrunus avium L.) has recently increased due to consumer awareness of their health benefits, as they are rich in polyphenolics   of fresh and dried SC was calculated using an oven at 105°C for 5 h (Shimaz).2.2To apply the microwave pretreatment on the SC, a microwave oven  was used under atmospheric pressure. In this work, the influence of the microwave pretreatment time at five levels of 0, 30, 60, 90, and 120 s (power = 220 W) on the SC was examined.2.3After each microwave pretreatment, the SC were dried in an oven , until reaching a constant weight.2.4Wt, We, and W0 are the WC of the SC at time t, equilibrium, and initial WC of the SC on a dry basis (g water/g dry matter). Data processing and modeling were performed using Matlab software (version R2012a) to estimate models parameters.The dehydration kinetics of SC have been explained using 10 simplified drying equations , r is the average radius of the SC, n is positive integers and t is drying duration (s). For food dehydration process modeling, Equation lnMDeff values from the slope of lnMR and t  plot, which is calculated by Equation Slo2.6R2 = 9998). The results were expressed as μg GAE/g dry matter. For the analysis of the antioxidant capacity, 2,2‐Diphenyl‐1‐picrylhydrazyl  free radical scavenging activity (FRSA) method was used according to Salehi, Ghazvineh, and Inanloodoghouz , all these data were adopted a one‐way ANOVA to determine differences in measured variables among experimental groups. Each treatment was conducted in triplicate and the results are expressed as the mean ± standard deviation .An analysis of variance (ANOVA) was conducted to analyze drying rate and physicochemical responses of SC according to the time of microwave exposure by SPSS 21 software (IBM). A post hoc test by Duncan's multiple range calculation at 33.1Dried fruits have a long shelf‐life, making them a good substitute for fresh fruits and allowing you to have off‐season fruits available. Hot air drying is the most common foods preservation method for fruits and vegetables Salehi, . The impp < .05 level between pretreatments (the time of microwave exposure). The drying duration of treated SC was shorter than the untreated samples. As the microwave pretreatment time enhanced from 0 to 120 s, the dehydration duration of SC in the hot air dryer significantly decreased from 220 to 150 min (p < .05). Sharma and Prasad . The purpose of the study by Simsek and Süfer  were obtained with freezing pretreatment, respectively. In another study, Simsek and Süfer  and the minimum SSE (sum of squared errors), and RMSE (root mean squared error) values  for all conditions compared to that of the other models. The calculated constant coefficients of the Midilli equation include a, k, n, and b, are reported in Table r) for all dehydration conditions. Mean values of SSE, RMSE, and r for all experiments ranged from 0.0002 to 0.0028, 0.0039 to 0.0154, and 0.9987 to 0.9999, respectively. Figure The drying behavior of untreated and microwave‐treated SC in the hot air dryer was fitted with the Midilli model . The average total phenolics of dried SC microwaved for 0, 30, 60, 90, and 120 s were 1491.4, 2127.8, 2127.2, 2330.7, and 2272.1 μg GAE/g dry, respectively. Cheng et al. . The antioxidant properties of the dried SC were retained by the microwave pretreatment. The antioxidant capacity of treated SC was higher than the untreated sample. The average DPPH FRSA of dried SC microwaved for 0, 30, 60, 90, and 120 s were 54.47%, 61.80%, 60.68%, 63.32%, and 62.59%, respectively. The impacts of edible coating and sonication on the total phenolic content and antioxidant capacity of SC were examined by Salehi, Ghazvineh, and Inanloodoghouz . This issue can be due to higher volume and porous structure with lower shrinkage in microwave‐treated samples which give higher diffusion of moisture inside the dried SC cells, and therefore, higher rehydration ratio. The average rehydration ratio of dried SC microwaved for 0, 30, 60, 90, and 120 s were 127.27%, 136.63%, 136.91%, 137.07%, and 136.72%, respectively. Nayi et al. . This may be due to less structural collapse during drying of microwaved SC, which may be due to the shorter heating time of microwaved SC. Considering the drying time, Deff values, total phenolics content, antioxidant activity, and rehydration ratio, microwave pretreatment will be more promising for SC pretreatment before drying process.In this work, the impacts of microwave pretreatment on the total phenolics content, antioxidant potential, and thin‐layer drying rates of SC were studied. The drying duration of microwave‐treated SC was shorter than the untreated sample. The Fakhreddin Salehi: Conceptualization (lead); data curation ; formal analysis ; investigation (lead); software ; supervision (lead); writing – original draft ; writing – review and editing . Moein Inanloodoghouz: Data curation ; formal analysis ; software . Sara Ghazvineh: Data curation ; formal analysis ; software .None.


However, an efficient, simple, and environmentally friendly method for separating V from its compounds is still lacking. In this study, we used first-principles density functional theory to analyse the vibrational phonon density of states of ammonium metavanadate and simulated its infrared absorption and Raman scattering spectra. By analysing the normal modes, we found that the V-related vibration has a strong infrared absorption peak at 711 cm−1, while other significant peaks above 2800 cm−1 are from N–H stretching vibrations. Therefore, we propose that providing high-power terahertz laser radiation at 711 cm−1 may facilitate the separation of V from its compounds through phonon–photon resonance absorption. With the continuous progress of terahertz laser technology, this technique is expected to be developed in the future, and it may offer new technological possibilities.Vanadium( −1 is from the V–O stretching vibrational mode. It is selected for ideal photon–phonon resonance absorption frequency.The peak at 711 cm It is resistant to both hydrochloric acid and sulfuric acid and outperforms most stainless steels in some ways. Because of its extensive utility, it is known as the metal ‘vitamin’. V is mainly used in the production of high-strength low-alloy steels, specialised steels, and aerospace alloys, as well as in other applications.1–3 In the field of aerospace, the excellent improvement effect of V in titanium alloys has been discovered, making it useful in body structures and jet engines.4–6 The global market for V redox flow batteries is also expected to grow at an annual rate of 59.7% between 2018 and 2025.7 Considering the production and demand data of V for the energy technology industry specifically, the demand for V is expected to increase by up to 73% by 2050.8 This dramatic increase will be due to the growing demand for new materials, making mining and recycling of renewable resources strategically important for many countries.Vanadium in 1950,21 many studies using infrared (IR) and Raman spectroscopy have focused on V in NH4VO3.22–30 Among these studies, Waal and Twu et al. investigated the pyrolysis process of NH4VO3 under the action of N2 and NH3 + H2O using Raman spectroscopy at the molecular level.24,27 However, there has been a lack of theoretical study of the lattice dynamic processes based on vibrational spectroscopy. In this work, we simulated the vibrational spectrum of NH4VO3 and analysed the normal modes. Through our analysis, we were able to assign V-related vibrational peaks and determine the IR-active modes of V in the NH4VO3 spectrum. Based on these findings, we propose a new method to assist V separation using photon–phonon resonance absorption (PPRA).Since the discovery of the crystal structure of ammonium metavanadate  exchange–correlation functional because the gradient of electron density in NH4VO3 varies widely.32 The convergence tolerance for energy and self-consistent field (SCF)was set to 1 × 10−9 eV per atom to eliminate virtual frequencies. The energy cut-off was set to 750 eV and the K-point mesh to 3 × 1 × 2 to calculate phonons using a norm-conserving pseudopotential and the linear response method. The property of polarisability, IR and Raman spectra were calculated. The simulated spectra can be compared with experimental data and the vibrational normal modes from the phonon calculation can be used for assignments.We performed geometry optimisation and phonon calculations using the CASTEP code,4VO3 has a pyroxene (Si2O6) structure, and its primitive cell contains 36 atoms with space group Pbcm.21,33,36 The crystal consists of four NH4+ clusters and two VO3− ribbons oriented in one direction. N3− is bonded in tetrahedral geometry to four H1+ atoms in NH4 clusters, while V5+ is bonded to four O2− atoms to form corner-sharing VO4 tetrahedra in each VO3 ribbon. In the analysis of vibrational modes, intermolecular vibrations were classified as translation and rotation modes, while intramolecular vibrations were classified as bending and stretching modes. The study focused on the V-related vibrational modes for further analysis and assignments.NH4VO3. Considering the normal vibrations at the gamma point, there are 108 normal modes for a 36-atom primitive cell, corresponding to 36 × 3 = 108 dispersion curves that make up the VDOS. Excluding three acoustic branches, there are 105 optical branches that can be detected by IR absorption or Raman scattering through phonon–photon coupling.4VO3, its IR- and Raman-active phonons are fully complementary. et al. reported the Raman spectrum of NH4VO3 in 1989 and assigned the peaks at 85.5, 149, 170, 195, and 209 cm−1 to translational modes, and the peaks at 269 and 311 cm−1 to rotational modes.36 Similarly, Du et al. reported the Raman and IR spectra and assigned the Raman-active peaks at 260 and 210 cm−1 to V–O bending vibrations,30 while Onodera et al. assigned the peaks at 205 and 255 cm−1 (Raman) and the peak at 223 cm−1 (IR) to V–O–V bending vibrations.34 In this work, we identified corresponding Raman-active vibrational modes at 44, 84, 137, 152, 195, 215, 261, and 312 cm−1, while the IR-active vibrational modes were at 99, 126, 172, 207, and 221 cm−1. In the low-frequency band, the vibrational modes are intermolecular collective vibrations. Our vibrational analysis confirmed that the modes at 44 and 84 cm−1 belong to NH4+ rotations, the modes at 99, 126, and 137 to VO3− rotations, the modes at 152, 172, 195, and 207 to NH4+ translations, and the modes at 215, 221, 261, and 312 cm−1 to skeletal vibrations, where the two ionic groups vibrate together. Due to the unique structure of NH−1, which are bending vibrations of the V–O bond. Onodera et al. compared NH4VO3 with KVO3 and assigned a peak at 375 cm−1 to the hydrogen bond vibration between NH4+ and VO3−.34 Du et al. attributed the mode at 385 cm−1 to the V PBM data was replaced with SVG by xgml2pxml:<glyph-data id="z.dbd" format="PBM" resolution="300" x-size="8" y-size="10" xml:space="preserve">00000000000000000000000000000000111111110000000011111111000000000000000000000000</glyph-data><svg xmlns="http://www.w3.org/2000/svg" version="1.0" width="13.200000pt" height="16.000000pt" viewBox="0 0 13.200000 16.000000" preserveAspectRatio="xMidYMid meet"><metadata>Created by potrace 1.16, written by Peter Selinger 2001-2019</metadata><g transform="translate scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg>O bending vibration.30 Park et al. discovered two Raman peaks at 326 and 344 cm−1, one of medium intensity and the other a shoulder peak.36 According to In et al. assigned various combined vibrations of V–O bonds between 500 and 850 cm−1 and a symmetric vibration of a Raman-active peak at 496 cm−1.30 Similarly, Onodera and Park assigned the Raman peak at 497 cm−1 to the symmetric vibration.34,36 They tentatively assigned the peaks using group theory. −1 corresponds to a skeletal rotation, as shown in In the IR spectrum, Du −1, which are all related to V–O stretching. Onodera and Park assigned three Raman peaks at 643, 895, and 925 cm−1, and three at 646, 897, and 936 cm−1, to asymmetric and symmetric V–O stretching vibrations, respectively. Onodera also observed IR peaks at 690, 850, and 935 cm−1. Du assigned two IR peaks at 895 and 935 cm−1 to V–O stretching. Our simulations are in good agreement with the experiments. We found five IR-active modes at 711, 862, 924, 975, and 994 cm−1, and five Raman-active modes at 712, 898, 948, 959, and 994 cm−1. Note that the IR-active mode at 994 cm−1 is not the same as the Raman-active mode at 994 cm−1. These two wavenumbers are not degenerate energies; they appear equal only due to number rounding. As shown in −1 correspond to the experimental IR peaks at 690, 850 cm−1 (ref. −1 (ref. −1 is presented in the ESI files.−1, the intensity ratios to 711 cm−1 are 45.55%, 17.23%, 17.02%, and 18.63%, respectively. It is clear that the IR peak at 711 cm−1 is very strong, which means that the PPRA effect at this peak should be very efficient. If a terahertz laser at this frequency is applied to NH4VO3, the V–O bonds will absorb the radiation energy efficiently, which may potentially break the chemical bonds and facilitate the separation of V from NH4VO3.There are 10 normal modes ranging from 700 to 1000 cmm−1 ref. , and 895 −1 ref. . The dyn−1, we found that all vibrations are related to the NH4+ group due to the low mass of the ions. Waal et al. attributed 1414 and 1422 cm−1 to the triply degenerate bending vibrations of NH4+ and pointed out a shoulder peak at 1406 cm−1.35 Onodera et al. suggested that the Raman peaks at 1420, 1440, and 1650 cm−1 represent NH4+ bending, while the peak at 1412 cm−1 represents NH4+ bending in the IR spectrum.34 Du et al. reported that the IR peak at 1416 cm−1 was related to the bending vibration of the N–H bond.30−1 all represent NH4+ bending.In the higher-frequency region ranging from 1418 to 3289 cm−1, the IR and Raman spectra exhibit several distinct characteristic peaks corresponding to N–H stretching vibrations. Waal et al. assigned vibrational peaks at 2839, 2926, and 3019 cm−1 to symmetric stretching, and peaks at 3122 and 3207 cm−1 to triply degenerate asymmetric stretching.35 Park et al. assigned Raman peaks at 2920 and 3050 cm−1 to NH4+ stretching.36 Onodera et al. observed IR peaks at 2800, 2980, and 3190 cm−1,34 while Du et al. assigned IR peaks at 2790, 2950 and 3200 cm−1 to NH4+ stretching.30 In this work, we identified that the modes at 2874, 2875, 2876, 3285, 3285, and 3289 cm−1 correspond to asymmetric stretching, while those at 2910, 2911, 2912, 2956, 3251, and 3253 cm−1 correspond to symmetric stretching. Although there are some high-intensity IR peaks in this range corresponding to N–H stretching modes, the PPRA effect does not directly facilitate V–O breaking. Two examples of normal modes at 1420 and 2876 cm−1 are shown in In the higher-frequency range above 2800 cm4VO3, we analysed the dynamic processes of the normal modes. The results show that the IR-active modes and Raman-active modes are fully complementary. Each vibrational normal mode is either IR-active or Raman-active.Based on density functional theory simulations of the VDOS of NH−1 represent the V–O stretching vibrations. The highest-intensity peak in the IR spectrum is at 711 cm−1, indicating that the PPRA effect of IR radiation at this frequency is very strong. Although there are still some high-intensity peaks in the region above 2800 cm−1, they do not stimulate the PPRA effect of V-related vibrations.In particular, we confirmed that the normal modes from 711 to 994 cm37 through metallurgical processing, where it is produced as a by-product. The V in the ore is usually in the form of powdered V2O3, which is then dissolved in water or an acidic or alkaline solution to form V-containing ion clusters.38–40 The two main chemical methods currently used for the industrial extraction of V are the sodium roasting–water leaching process9–13 and the calcium roasting–acid leaching process.14–18 Based on our mode analysis, we propose the use of a high-power terahertz laser radiation at 711 cm−1 to assist in breaking the V–O bonds and separating V from NH4VO3. With the continuous progress of terahertz laser technology, this PPRA method could offer new application prospects. By utilising this PPRA physical method, it may be possible to achieve an environmentally friendly and efficient extraction of V from ores.V is typically obtained from V-bearing titanomagnetite and ilmenite oreThere are no conflicts to declare.RA-013-D3RA02053C-s001


A. arguta (Actinidia arguta) is an anthocyanin-rich and excellent hardy fruit. Many studies have focused on the green-fleshed A. arguta, and fewer studies have been conducted on the all-red A. arguta. Here we reported a regeneration and Agrobacterium-mediated transformation protocol by using leaves of all-red A. arguta as explants. Aseptic seedling leaves of A. arguta were used as callus-inducing materials. MS medium supplemented with 0.3 mg·L-1 2,4-D and 1.0 mg·L-1 BA was the optimal medium for callus induction of leaves, and medium supplemented with 3 mg·L-1 tZ and 0.5 mg·L-1 IAA was optimal for adventitious shoot regeneration. The best proliferation medium for adventitious buds was MS + 1.0 mg·L-1 BA + 0.3 mg·L-1 NAA. The best rooting medium was 1/2MS + 0.7 mg·L-1 IBA with a 100% rooting rate. For the red flesh hardy kiwi variety ‘Purpurna Saduwa’ (A. arguta var. purpurea), leaves are receptors for Agrobacterium (EHA105)-mediated transformation. The orthogonal experiment was used for the optimization of each genetic transformation parameter and the genetic transformation of the leaves was 21% under optimal conditions. Our study provides technical parameters for applying genetic resources and molecular breeding of kiwifruit with red flesh.The all-red  Actinidia arguta) is native to China and is abundant in locations such as the northeast and north of the country. It has strong resistance to adverse conditions and is an excellent rootstock material. As a third-generation emergent dual-purpose fruit, it not only has a distinct fruit flavor and is easy to consume, but it also has a large market prospective and development value  is a Ukrainian-bred variety, with red flesh and skin , IBA (Indole-3-butyric acid), and IAA (Indole-3-acetic acid) were used as auxins  HgCl2 for 6 min and washed three times with sterile water. Next, the branches were cut into 3-cm long stem segments with buds, and the morphological lower ends of the explants were inoculated onto MS + 1.0 mg·L-1 BA + 0.2 mg·L-1 NAA medium. Five explants were inoculated per bottle and placed in the culture room for 30 d to obtain aseptic plants.PGR (Plant growth regulator) ratios of tZ (Trans-Zeatin), TDZ (Thidiazuron), NAA, and IAA, were cultured in the dark for 30 d and then under a 16 h light/8 h dark photoperiod for 30 d. The culture-induced adventitious shoots, about 3 cm in height, and these were transferred to MS medium without PGRs for 15 days, and then transferred to MS medium containing different hormone ratios of BA and NAA for the proliferation of adventitious shoots for 30 days under a 16-hour photoperiod. Rootless plantlets  were inoculated on 1/2 MS medium with IBA  for root induction. All media were solidified with 7.0 g·L-1 agar, supplemented with 30 g·L -1 sucrose, and the pH was adjusted to 5.8. All experiments were repeated three times, treating ten explants at a time. And all experiments were performed at 25 ± 2°C and with a 16 h/day photoperiod with 30 µE m-2 of photosynthetically active light.Aseptic leaves (5 mm x 5 mm) were inoculated on MS  medium w-1 BA, 0.3 mg·L-1 2,4-D, 300 mg·L-1 Timentin, 300 mg·L-1 Carb (Carbenicillin), and various doses of Kan (Kanamycin)  and Hyg (Hygromycin B) , in order to determine the optimal Kan or Hgy concentration for selecting transformed callus. In each Petri dish, 10 leaf discs were grown at each Kan and Hgy concentration, with three duplicates per dish. After 30 days of culture, the growth and development situation and survival rate of explants were recorded.The differentiating capacity was evaluated using 5 mm × 5 mm leaf pieces. Leaf pieces were cultured in media containing 1.0 mg·LThe Agrobacterium tumefaciens strain was EHA105  and was stored at -80°C. The transformation plasmid was pCAMBIA1301, which contained the CaMv35S-regulated 1,3-glucosidase (GUS) reporter and the neomycin phosphotransferase (NPT‖) gene against kanamycin (Kan). An intron was inserted between the CaMv35S promoter and the GUS gene, which is expressed solely in plant cells.Agrobacterium was incubated in a YEP liquid medium containing antibiotics at 28°C for 36–48 h on a constant temperature shaker at 220 rpm. Then, the bacterial culture was incubated until it appeared clear yellow. Next, 100 mg·L-1 AS (Acetosyringone) was added at the appropriate concentration, and the bacterial culture was further incubated for 4 h until OD600 = 0.6–0.8. Finally, the Agrobacterium bacterial culture was centrifuged at 3,000 rpm for 10 min and the supernatant was discarded. A volume of 30–40 mL sterilized distilled water was added to the centrifuge tube. The bacteria were resuspended by gently aspirating with a pipette. Next, the bacterial culture was centrifuged at 7,000 rpm for 5 min at room temperature, this step was repeated once. Finally, the supernatant was poured off, and then about 50 mL of MS liquid medium was added to the centrifuge tube, and it was set aside.PGRs for 30 d were selected, and the lower middle part of the leaf (Pre-experiments showed a high regeneration rate in the lower and middle part of the leaf) was cut into pieces of 5 mm x 5 mm with scissors. The leaves were inoculated on MS medium containing 1.0 mg·L-1 BA and 0.3 mg·L-1 2,4-D and placed in the dark for pre-culture . Bacterial culture of different OD600  were added to the Petri dishes followed by pre-cultured leaves for infestation  with intermittent shaking during the period. Afterward, the bacterial solution was poured off, the excess bacterial solution was blotted off the leaves with sterile filter paper, and the leaves were placed on MS medium containing 1.0 mg·L-1 BA, 0.3 mg·L-1 2,4-D, and appropriate concentrations of AS  and co-cultured for 3 d at 25°C in the dark. The leaves were transferred to MS medium containing 1.0 mg·L-1 BA, 0.3 mg·L-1 2,4-D, 300 mg·L-1 Timentin, and 300 mg·L-1 Carb for delayed screening . Afterwards, leaves were transferred to MS medium containing 3.0 mg·L-1 tZ, 0.5 mg·L-1 IAA, and 10 mg·L-1 Hyg for resistance screening. Subculturing was performed every fifteen days and collected after a total of three subculturing. One treatment was repeated five times, for a total of 100 leaves.Invitro-raised seedlings grown on MS medium without After incubation on the selection medium for >45 d, selected resistant calli surviving in the medium were immersed in a GUS staining solution  and held at 37°C for 12 h. The callus samples were decolorized by transferring them into 70% ethanol 2–3 times until the negative control material was white. Successfully stained calli were photographed, and the data was recorded. The genomic DNA was extracted using a Plant DNA Isolation Kit . GUS-forward (5’-AACCGACGACTCGTCCGTC-3’) and GUS-reverse (5’-GAAGTTCATGCCAGTCCAGCG-3’) primers were used to detect exogenous GUS insertions. PCR was performed using 2×M5 PAGE Taq PCR Mix  according to the manufacturer’s instructions, and the PCR products were analyzed by agarose gel electrophoresis.P<0.05). All data passed the homogeneity test before Duncan’s multiple range test was applied, and Microsoft Excel software was used to analyze and display the results.IBM SPSS Statistics 20.0 was used to identify significant differences between treatments  and highest fresh weight (9.85 g) was on the medium of 1.0 mg·L-1 BA + 0.3 mg·L-1 2,4-D. Moreover, as seen in A. arguta leaves. In this study, it was concluded that the best combination was 1.0 mg·L−1 BA + 0.3 mg·L−1 2,4-D (As shown in -1 tZ + 0.1 mg·L-1 NAA at 43.33% and 10.3, respectively. When the auxin was IAA under the same tZ concentration conditions, the regeneration rates of both low and high IAA concentrations were not significantly different and did not change significantly. In the experiment, it was also noted that when IAA was used as the auxin, sprouting occurred earlier than when NAA or no auxin was used. On the medium containing IAA, visible buds appeared after one week of incubation in the light and expanded swiftly into visible seedlings as the incubation period rose, though on the medium containing NAA, visible buds appeared only after two weeks of incubation in the light, and their growth was sluggish. When the tZ concentration was varied over a relatively low range, neither the regeneration rate nor the number of adventitious shoots significantly changed. However, when the tZ concentration was increased to 3 mg L-1, both the regeneration rate and the number of adventitious shoots grew substantially. Therefore, the ideal hormone combination for adventitious shoot regeneration was 3 mg·L-1 tZ + 0.5 mg·L-1 IAA.According to The effect of TDZ on the regeneration of adventitious shoots was not significant. The regeneration rate of leaf adventitious shoots was higher for low concentration of TDZ than for high concentration of TDZ. It was observed in the experiment that the differentiation of leaf adventitious shoots was late, and most of them started to appear buds only 8 weeks after inoculation. Meanwhile, the induced adventitious shoots were basically single shoots, and extended incubation time revealed that most of the shoots eventually failed to grow into a robust seedling. Therefore, TDZ was found to be unsuitable as a cytokinin for the regeneration of ‘Purpurna Saduwa’ adventitious shoots in this experiment.The leaves were cultured on a regeneration medium for about 15 d (A. arguta were beginning to grow from the axillary buds (During the same experiment, it was noticed that adventitious shoots of -1, the maximum number of roots was 17.60, but the rooting rate was only 90%. When IBA was 0.3 mg·L-1 and 0.7 mg·L-1, the rooting rate reached 100%, and the number of roots was 5.90 and 14.50, respectively, and the average number of roots of the latter was significantly higher than that of the former. The fresh weight of shoots also increased with the increase of IBA concentration but in the treatment of IBA at 1.0 mg·L-1 (-1.Four days after inoculation, the shoots began to generate callus at the base. A few shoots began to root at seven days. The majority of shoots began to root around 10 days, and after 20 days, no new roots emerged, the roots only became longer (After acclimatization, the plantlets were withdrawn from the vials, the roots were rinsed, and the plantlets were transplanted into a substrate consisting of 7:3 coconut coir: perlite. The pots were properly watered with tap water for the first time. Trays with a 1,000-fold diluted nutrient solution were placed beneath the pots, and the pots were covered with a moisturizing lid to keep them hydrated. A total of 30 seedlings were transplanted and they all survived and flourished (-1, no callus growth was observed on the leaves. As the concentration of Kan was raised, neither callus growth nor browning death was noticed on the leaves. It was thus impossible to determine the deadly concentration of Kan on the leaves. When the antibiotic was substituted with Hyg, the leaves were discovered to be more susceptible to Hyg, with no callus formation at 5 mg·L-1 and browning and death of the leaves at 10 mg·L-1. Therefore, 10 mg·L-1 Hyg was the fatal concentration for leaves and 10 mg·L-1 was chosen as the resistance screening threshold concentration.600.If a single-factor design is used, the best single-factor values are selected one by one, so that the theoretical best combination of conditions selected item by item is not necessarily the true best combination of conditions. If an orthogonal experiment is used, it not only allows for a more scientific experimental design, but also has a balanced dispersion of tests and simple data calculation, as well as reduces workload. As depicted in -1 AS was added to both the culture broth and co-culture medium. Therefore, the optimum AS concentration was 100 µmol·L-1. As shown in 600 of 0.2 and 0.8 was significantly lower than that for 0.4 and 0.6. When OD600 was 0.4, the mean genetic conversion rate was 7.25%, when OD600 was 0.6, the mean genetic conversion rate was 7.50%, and there was no difference between these two genetic conversion rates. Therefore, the optimal OD600 was 0.4 to 0.6.The optimum level for each treatment was derived by range analysis. As can be seen from 600 of 0.4–0.6, infection for 20–30 min, the addition of 100 µmol·L-1 AS to the culture broth and co-culture medium, co-culture for 3 d, and delayed culture screening for 0–2 d.In summary, the optimal conditions for genetic transformation were: pre-culture for 4 d, ODThe vector carried the GUS gene, thus, the resistant calli was stained with GUS. Those that were fully stained blue were positive and those that were not stained blue were negative, and the genetic transformation rate was determined based on this staining data and PCR identification. The staining results are presented in −1, browning was severe in all three treatments of NAA, indicating that browning was aggravated when BA concentration was too high. Liu et al. found that both high concentrations of BA and low concentrations of NAA aggravated the browning of explants during tissue culture of A. arguta, which is similar to the findings of the present study (A. arguta callus induction.In plant tissue culture, the type of plant growth regulator and the concentration ratios employed are crucial criteria for experimental success . In thisnt study . 2,4-D int study . Wang etA. arguta plants. Meanwhile, TDZ, as a highly potent cytokinin, has a better effect on the regeneration systems of strawberries and pears. A. deliciosa, a finding contrary to the present study, probably due to the difference between the culture and the plant itself. In the present study, it was also discovered that plants could be regenerated only on a medium containing tZ. Therefore, auxin only plays an auxiliary role in the differentiation of explants, and IAA is more effective than NAA. A. arguta regeneration system, and its effect is irreplaceable by TDZ.Trans-Zeatin and BA are the most commonly-used cytokinins for the induction of kiwifruit leaf regeneration, but the appropriate cytokinins in kiwifruit regeneration vary among genotypes. In particular, tZ is frequently used for adventitious shoot induction in kiwifruit callus . TransZe−1 during the proliferation of adventitious shoots. This observation indicates that ‘Purpurna Saduwa’ is more sensitive to BA and can tolerate lower concentrations of BA. -1 and NAA mg·L-1 showed the highest proliferation rate of 2.91 for axillary buds. −1 IBA, resulting in a rooting rate of 94%. In the ‘Purpurna Saduwa’, it showed a higher propensity for rooting at an IBA concentration of 0.7 mg·L−1, which aligns with previous research findings. The best medium for rooting was 1/2MS + 0.5 mg·L−1 IBA, with a rooting rate of 94%. The rooting of ‘Purpurna Saduwa’ was easier and the best IBA concentration for rooting was 0.7 mg·L−1, which is generally consistent with previous studies (A. arguta has good economic benefits and broad market prospects. With the rise of industrialization of A. arguta cultivation, the demand for this high-quality kiwi species has gradually increased, and the establishment of a regeneration system for A. arguta is a key solution to this problem. However, the genetic background of A. arguta is complex and plant regeneration is difficult. Regeneration systems have been established on only a few kiwi varieties, and the regeneration rates obtained are not high, and stable and efficient regeneration systems are lacking. Through this study, an effective method to improve the quantity and quality of A. arguta reproduction was explored. The resulting findings contribute to developing a fast-breeding system for A. arguta, provide a theoretical basis for the factory breeding of all-red A. arguta plantlets, and offer a way to reduce the breeding cost. A. arguta, therefore, may become better used in the food and medicine industries and provide a large amount of high-quality material for research on the genetic transformation system of A. arguta as well as its disease resistance, micrografting technology, etc.In this experiment, it was found that deformed plantlets easily appeared when the BA concentration was 2.0 mg·L studies . As a smPlant transformation plays a key role in plant improvement by introducing beneficial exogenous genes or by silencing the expression of endogenous genes in crop plants . TransgeA. arguta leaf tissue was created.The Agrobacterium tumefaciens-mediated genetic transformation system is currently the most desirable transformation method for dicotyledonous plants. Its transformation efficiency is often influenced by strain type , explant-1 Hyg may prevent the development of non-transformed tissue in grapes (-1 of Hyg was the efficient concentration for screening positive callus. This research confirms that pre-culturing for four days can put the explant cells in a state where they are easily able to receive information, which is conducive to the transformation of Agrobacterium and can also reduce the damage caused by Agrobacterium to the explant, thereby increasing the transformation efficiency. This is consistent with the prior findings of kiwifruit genetic transformation studies (600 value was between 0.4 and 0.6, the infection time was between 20 and 30 minutes, and the addition of 100 mol·L-1 AS during the Agrobacterium activation stage and co-cultivation improved transformation efficiency, the greatest rate of expression was 21%. In this work, it was also proven that adding AS during the activation stage of Agrobacterium and co-cultivation may considerably enhance transformation efficiency. As infection materials, -1 AS to the Agrobacterium activation medium and the co-culture medium simultaneously increased the transformation efficiency. -1 AS to the bacterial solution increased the transformation efficiency of Geranium notably more than other concentrations. In this study, it was noticed that the callus development of infected explants was sluggish. The callus structure was soft, pale in color, and its process of redifferentiation was challenged. Consequently, no transgenic plants were obtained throughout this research. Kan and Hyg are commonly utilized as screening antibiotics in kiwifruit genetic transformation. Compared to Kan, Hyg inhibits plant development more obviously. For instance, 2–10 mg·Ln grapes , pear (Xn grapes , etc. In studies . The OD6A. arguta transgenic plants with GUS by infecting A. arguta‘K2D2’with EHA105 and by reducing the basal salt strength of the medium, based on the results of A. arguta. A. deliciosa, A. chinensis, A. eriantha Benth, and A. arguta and demonstrated that the transformation rate and a number of transgenic plants obtained were lowest in A. arguta and significantly lower than in the other three species. A. arguta and verified the anthocyanin synthesis function of the gene. However, the process did not have a high rate of shoot regeneration, and the genetic transformation efficiency was low. A. arguta was strongly influenced by variety, with significant differences in regeneration rates between varieties. Most of the varieties studied for these studies were green-fleshed A. arguta (A. arguta. Due to the complicated genetic background of A. arguta and the difficulties of plant regeneration, regeneration systems have only been established in a few varieties, and the regeneration rates obtained are low, indicating the lack of a stable and efficient regeneration system. In addition, research on genetic transformation in A. arguta has only been conducted for a short length of time. As a result, there is a lack of a stable and comprehensive genetic transformation system. This experiment innovatively established a regeneration system and genetic transformation system for the all-red A. arguta. All transplanted plants survived, which is important for the conservation of germplasm resources and practical production applications.. arguta . However. arguta . These aThe original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.WY: Writing - original draft, Visualization, LK: Investigation, Methodology, Data curation, DL: Investigation, Validation, BZ: Methodology, Validation, HT: Software, Methodology, XZ: Investigation, Methodology, YuL: Formal analysis, YTZ: Investigation, QC: Software, YW: Visualization, ML: Investigation, WH: Investigation, YaL: Visualization, XW: Visualization, HRT: Project administration, YZ: Writing - review and editing, Conceptualization, Funding acquisition. All authors contributed to the article and approved the submitted version.