This study found that the grain yield of the tested rice cultivars ranged from 5.75–11.00 t ha−1, with a growth duration lasting between 144–161 days across the cultivars. These cultivars were then categorized into high yielding (HY), medium yielding (MY), and low yielding (LY) based on daily yield by using Ward’s hierarchical clustering method. The higher grain yield for HY compared to MY and LY was attributed to more spikelets per unit area. The HY alleviated the conflict between higher panicle density and larger panicle size by improving the seedling emergence rate and productive stem rate. It also significantly increased shoot biomass at maturity. The HY reduced the period between seeding and beginning of heading (BBCH 51) and the proportion of dry matter partitioned to the leaf at the heading stage. However, it also increased the accumulation of dry matter in the grain and the proportion of dry matter partitioned to the grain at maturity. Furthermore, the HY markedly increased the harvest index and grain-leaf ratio, which are beneficial to coordinate the source–sink relationship. A quadratic function predicted that 98 days is the optimum growth duration before heading (BBCH 51) for achieving maximum yield. In conclusion, for dry direct seeding rice, it is appropriate to select high-yielding japonica inbred rice cultivars with shorter growth duration before heading (about 93–102 day), higher panicle number (about 450–500 × 104 ha–1), more spikelet number per panicle (about 110–130), higher seedling emergence rate (about 70–75%), higher productive stem rate (about 60–70%), and greater harvest index (about 50–55%).Dry direct seeding rice (DSR) is an emerging production system because of increasing labor and water scarcity in rice cultivation. The limited availability of rice cultivars suitable for dry direct seeding hampers the widespread adoption of this cultivation method in Northeast China. This study aimed to investigate grain production and plant characteristics associated with dry direct seeding rice. We conducted a field experiment on 79  Oryza sativa L.) is the primary crop and constitutes a staple diet for over half of the global population [Rice  to evaluate the yield and yield components of dry direct seeding rice of the three different cultivar types; (2) to measure the production of dry matter, distribution, and source–sink characteristics of the different cultivar types; and (3) to identify the relationships among growth duration and grain yield. The results of this study might offer both theoretical and empirical foundations for the selection and breeding of rice cultivars with high-yielding that are appropriate for dry direct seeding systems.Therefore, a field experiment was carried out in Shenyang City, Liaoning Province, China in 2020 and 2021. This study categorized 79 −1, 27.48–70.98 kg ha–1 d–1, and 144–161 d. The selected rice cultivars were separated into three cultivar types, including high yielding (HY), medium yielding (MY), and low yielding (LY), using Ward’s method for hierarchical clustering  . HY had Cultivar type markedly affected stem, leaf, and panicle biomass amount at each growth stage . HY and The proportion of dry matter partitioning to the leaf markedly decreased after the heading stage . In contDifferences were observed amongst cultivars in both the percentage of exported matter and the percentage of matter transformed in the stem-sheath, as demonstrated in Cultivar type markedly affected the LAI, SPAD, and HI . RelativGrain yield and daily yield increased significantly with an increase in panicles per unit area, which showed a close correlation with both seedling emergence rate and productive stem rate . Grain yThe regression model between grain yield and the period from seeding to beginning of heading (BBCH 51) (X) for high-yielding cultivars was established using the method of polynomial regression analysis , and theAccording to the regression equation, there was a single-peak curvilinear relationship between grain yield and growth duration before heading (BBCH 51) for HY cultivars. The grain yield first increased and then decreased with delaying the growth duration before heading (BBCH 51). Based on this quadratic equation, the maximum grain yield of rice cultivars suitable for dry direct seeding rice system may be obtained when the period from seeding to beginning of heading (BBCH 51) of rice cultivars were 98 d.japonica inbred rice cultivars that have differing yield levels. Moreover, we identified certain selection criteria for high-yielding rice cultivars.Dry direct seeding rice is a promising option for rice production in China and other countries, based on less water resource input and higher production efficiency ,34. Curr−1. We segregated these cultivars, based on the results of the hierarchical cluster analysis, into high-yielding (HY) cultivars, medium-yielding (MY) cultivars, and low-yielding (LY) cultivars. The significant difference in grain yield can be attributed to the variation in spikelets per unit area.  because HY had a higher grain–leaf ratio. Therefore, the rice population of HY cultivar showed a coordinated enhancement of sink size and source capacity.The formation of crop yield is a process of source–flow–sink interaction and is related to the production, transport, and accumulation of photosynthate ,45. SourIntercepted solar radiation during the growing season is a significant driver of crop canopy photosynthesis and biomass production ,47. The −1 of organic matter, 1.52 g kg−1 of total nitrogen, 107.42 mg kg−1 of available nitrogen, 15.84 mg kg−1 of available phosphorus, and 92.36 mg kg−1 of available potassium.Field trials were conducted in a farmer’s field located in Shenyang City, Liaoning Province, China (42°24′ N 122°59′ E), in 2020 and 2021. Shenyang has a temperate semi-humid continental climate. Between April and September 2020, the average ambient temperature and precipitation were 20.09 °C and 87.62 mm, respectively. Between April and September 2021, the average ambient temperature and precipitation were 19.69 °C and 119.16 mm, respectively. Before plowing, soil samples were collected from the assessed fields at a depth of 0–20 cm. The sample was air-dried, sieved, and analyzed in the laboratory using standard methods to estimate the physical and chemical properties. The experimental site soil texture was loam with a pH of 5.77, 14.45 g kgjaponica inbred rice cultivars were used as experimental materials . The seeding rate was 90 kg hm−2 (390 seeds per square meter), and the sowing depth was 2 cm. The total N amounts of the N fertilizer application rate was 190 kg ha−1, which was sequentially applied as follows: 40% at basal, 30% at tillering stage, 20% at panicle initiation stage, and the remaining 10% at the heading stage of each cultivar. All nitrogen fertilizers were applied in the form of urea. Phosphorus (100 kg P2O5 ha−1 as triple superphosphate) was applied in all plots one day before sowing. Potassium (75 kg K2O ha−1 as potassium sulfate) were equal applied in all plots one day before sowing and at panicle initiation stage, respectively.Field experiments were conducted with a completely random block design with three replications. The pretreated rice seeds were sown on 26 April 2020 and 28 April 2021, respectively. Fields were rotary tillage for 15 cm depth, suppression, then rotary tillage for 5 cm depth, harrowed, without peddling, and leveled before sowing. The size of each plot was 18 mWater management: All treatments were flooded after sowing to maintain the soil moisture level and then wetting–drying alternation irrigation was performed after seedling emergence. A flood water depth of 3–5 cm was maintained after the four-leaf stage. Water was then drained at the maximum tillering stage to reduce unproductive tillers and re-watered at the panicle initiation stage with a 3–5 cm layer of water until the heading stage. Wetting–drying alternation irrigation was performed during the grain filling stage, and the water was drained 10 days before maturity.−1), 75% Halosulfuron-methyl GR (56.25 g a.i. ha−1), 25 g/L Penoxsulam SC (22.5 g a.i. ha−1), and 30% Cyhalofop-butyl SC (225 g a.i. ha−1). Herbicides were applied twice, one using 480 g/L Clomazone EC (252 g a.i. ha−1), and 75% Halosulfuron-methyl GR (56.25 g a.i. ha−1) within three days after sowing, the other at six-leaf age using 25 g/L Penoxsulam SC (22.5 g a.i. ha−1) and 30% Cyhalofop-butyl SC (225 g a.i. ha−1). All herbicides were sprayed manually with a 3 WBD-18 knapsack electric sprayer. The sprayer was cleaned repeatedly before and after each application. After the second herbicide application for one day, a 3–5 cm water layer was maintained in the field for 5 days, and, during this period, the water layer was lower than 2/3 of the stems and leaves of rice.Weed management: 480 g/L Clomazone EC  and productive stem rate (PSR) were calculated as follows:The dates of sowing, heading stage, and maturity were recorded to determine the growth duration (GD). The seedling emergence rate was investigated 21 days after direct seeding. An area of 0.5 m2 from each plot were selected for sampling. Following the recording of the number of panicles, the plant samples were separated into leaves, stems (culm plus sheath), and panicles. The dry weight of each organ was determined after oven-drying at 80 °C to achieve a constant weight. The total biomass was defined as the sum of the dry matter weight of the organs. The biomass at heading stage (BHS) and biomass at maturity stage (BMS) were used to represent the total biomass of each stage, respectively. The percentage of dry matter accumulations in the organs were calculated. The export percentage of the matter in stem-sheath (EPMSS) and the transformation percentage of the matter in stem-sheath (TPMSS) were measured as follows:At the heading and maturity stages, plants with an area of 0.5 mDuring the heading stage, a leaf area meter  was used to measure the green leaf area of the plants and determine the leaf area index (LAI). The SPAD measurements were conducted using the SPAD-502 Chlorophyll Meter . To present the source–sink relationship of rice, we used the grain–leaf area ratio, including three forms, spikelet-leaf area ratio (SNR), grain–leaf area ratio (GNR), and grain weight–leaf area ratio (GWR), which were calculated as follows: 2 area at maturity. Each sample was counted for panicle number to determine the effective panicles (EP) per square meter, and the plants were then separated into straw and panicles. The grains were manually separated from the straw by threshing the plants. The grain samples, which had been threshed, were air-dried and then submerged in tap water to separate filled and unfilled spikelets. They were then oven-dried at 80 °C to a constant weight. Three subsamples, each containing 20 g of filled grains, were manually counted to determine the 1000-grain weight. First, we divided 20 by the number of counted filled spikelets and then multiplied the result by 1000. Next, we calculated the number of filled spikelets by dividing the total dry weight of filled grains by the grain weight. We manually counted all of the unfilled spikelets and calculated the total number of spikelets per square meter (SPM) including both filled and unfilled ones. We calculated the spikelets per panicle by taking the ratio of the total spikelet number to the panicle number. Additionally, the seed-setting rate was calculated by taking the ratio of the filled spikelet number to the total spikelet number [2 (the middle five rows by 5 m) area of each plot with a sickle. The grain yields (GY) were converted to a moisture content of 14.5%. The daily yield (DY) was the ratio between grain yield per unit area to the whole duration days. The harvest index (HI) was the ratio between gain yield to aboveground biomass.To gather data on rice yield and yield components, plants were sampled from a 0.5 mt number . The graThe calculated daily yield of each of the 79 cultivars was used to classify them into different categories using Wards’ method of hierarchical cluster analysis. This analysis was conducted through the usage of the R (R 4.2.2) ‘stats’ package and ‘ggtree’ package. We used the least significant difference (LSD) test at a 0.05 probability level to compare the differences between treatments or multiple comparisons, using SPSS v. 26 . We used the Pearson correlation coefficient to analyze the relationship between various traits, at either the 0.05 or 0.01 probability levels. We used Origin version 2022  for the graphical illustration of the data. To establish the regression models between grain yield and growth duration of the heading stage, we used polynomial regression analysis.japonica inbred rice cultivars were classified into high-yielding (HY), medium-yielding (MY), and low-yielding (LY) cultivar. The HY had greater aboveground biomass and harvest index, mainly due to higher dry matter accumulation after heading stage. HY also increased the post-heading stage dry matter production and the accumulation of dry matter stored in the grains. In addition, the HY significantly enhanced both seedling emergency rate and productive stem rate, aiding in the conflict reduction between larger panicle size and higher panicle number per unit area. Furthermore, the HY cultivars coordinated the source and sink relationship by markedly improving grain–leaf ratio. Thus, HY enhanced the amounts of spikelets per unit area, leading to a 29.74–74.85% higher grain yield compared to MY and LY. Thus, rice cultivars with comparatively greater seedling emergency rate, productive stem rate, larger panicles, higher dry matter accumulation after heading stage, and higher apportionment of total dry matter towards grains at maturity stage can be chosen as HY japonica inbred rice cultivars for direct seeding cultivation.Ensuring food security and high production efficiency are important prerequisites for achieving the transformation of rice cultivation system. In this study, 79 tested


P < 0.0001). Among the components of MetS, significant associations were found between BD and diabetes mellitus , BD and hypertension , and BD and dyslipidemia . Our study indicated an association between BD and the risk of MetS and some of its components . Physician should consider these associations so that specific treatments are available for patients with comorbidities. Moreover, patients with BD should regularly monitor their blood pressure, fasting plasma glucose, and blood lipid levels.The present meta-analysis aimed to elucidate the association of Behçet’s disease (BD) with the risk of metabolic syndrome (MetS) and its components. Observational cohort studies were searched from the Embase, Web of Science, Medline, and Cochrane Library databases. The primary outcome was the association of BD with the risk of MetS and its relevant components. Effect estimates with odds ratios (ORs) were pooled using either the random-effects or fixed-effects models, according to heterogeneity. Leave-one-out sensitivity analyses were used to determine the stability of the results. Twenty-three studies, comprising 42,834 patients with BD, were included. Overall, a significant association between BD and the risk of MetS was found (pooled OR 2.26; 95% confidence interval [CI] 1.61–3.17; The online version contains supplementary material available at 10.1007/s10238-023-01044-x. Behçet’s disease (BD) is a multisystemic autoimmune and chronic inflammatory vasculitis, characterized by recurrent painful mouth sores, genital ulcers, uveitis, skin lesions, and other systemic manifestations . It freqMetabolic syndrome (MetS) affects 14–32% of the world’s population, and its incidence continues to increase . MetS isRecent studies have reported that patients with BD are more likely to have MetS than health control (HCs) –16; howehttp://www.prisma-statement.org/) guidelines [This study adhered to the Meta-analysis of Observational Studies in Epidemiology (MOOSE) and the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA)  searched for studies published before November 31, 2022, using electronic databases . The terms used included Behçet syndrome, Behçet disease, Behçet’s syndrome, Silk syndrome, Behçets syndrome, Behçet’s disease, BD, metabolic syndrome, metabolic disorders, hypertension, blood pressure, fasting blood glucose, plasma glucose, dyslipidemia, triglyceride, HDL, waist circumference, obesity, and abdominal obesity. We also performed a manual supplemental search by reviewing the reference lists of relevant articles, systematic reviews, and meta-analyses to avoid potentially missing articles. The review protocol was registered in the International Prospective Register of Systematic Reviews (PROSPERO) (CRD42022344815).The inclusion criteria for studies in the meta-analysis were as follows: (1) prospective or retrospective observational studies; (2) studies involving human participants; (3) studies investigating the association of BD with MetS or its relevant components, and studies describing the prevalence of MetS or its relevant components in BD patients; and (4) studies published in English.The primary outcome was the association between BD and the risk of MetS and its relevant components.Two authors (TC and XS) independently engaged in the study selection, data collection, and extraction. In the case of incomplete data, we emailed the authors to obtain supplementary information. The quality of each eligible study was assessed using the Newcastle–Ottawa scale (NOS) by each investigator to evaluate the quality . The NOSI2 test; I2 > 50% indicated that the studies were heterogeneous. If considerable heterogeneity (I2 > 50%) was recognized, the random-effects model was employed (DerSimonian and Laird method); otherwise, a fixed-effects model was used (Mantel–Haenszel method). Begg’s test was conducted to evaluate publication bias [All statistical analyses were performed using Review Manager 5.4 software  and the Bioconductor programming environment 22 . Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to assess the prevalence of MetS and its components in the comparison between patients with BD and controls. Statistical heterogeneity between studies was calculated using the ion bias , 21.The literature search process is illustrated in Fig. P < 0.0001; Fig. I2 = 27%; P = 0.21). Additionally, we did not examine publication bias based on the symmetry of the funnel plot and the Begg’s test (P = 0.4579).The prevalence of MetS in patients with BD was reported in eight studies , 22–27. P < 0.0001), and the heterogeneity of these studies was evident . The prevalence of diabetes mellitus in patients with BD using the Begg’s test (P = 0.3252) revealed no publication bias. Moreover, seven studies that reported fasting blood glucose levels in patients with BD were included, and no significant difference in fasting blood glucose was observed  [Fifteen studies reported the prevalence of diabetes mellitus in patients with BD, including 10 case–control studies, three cohort studies, and two cross-sectional studies. The pooled prevalence of diabetes mellitus in patients with BD was 11%  , 34, 35.P = 0.04; Fig. I2 = 61%; P = 0.03) [P = 1.0000).As illustrated in Fig.  = 0.03) , 35, 37.P = 0.11; Supplementary Fig. 2). A lower HDL level was observed in patients with BD in most studies; however, the difference was not markedly significant .Twelve studies recorded triglyceride levels , 35, 38,P = 0.002), with considerable heterogeneity . Of these studies, two recorded systolic and diastolic blood pressure (DBP) levels in patients with BD [P = 0.11); however, an evident difference in DBP between patients with BD and HCs  was observed .The results of the nine trials including 35,648 patients, also demonstrated an increased OR for hypertension in association with BD , 32–38.  with BD , 24. TheP = 0.90; Supplementary Fig. 5A) [P = 0.59; Supplementary Fig. 5B), or between BD and waistline .The pooled analysis of ORs of two individual studies showed no significant association of BD with obesity  , 35. ThiFig. 5A) –36, 38, Fig. 5A) , 33, 38 P = 0.16), with substantial heterogeneity . We conducted sensitivity analyses for BD and insulin resistance; the results did not show any significant alteration in the pooled OR when any individual study was sequentially omitted, demonstrating its stability and credibility. No evidence of potential publication bias was shown using Begg’s test (P = 0.4969). A subgroup analysis performed primarily based on different diagnostic tools of included trials revealed that patient with Behçet disease was associated with elevated incidence of insulin resistance when the diagnostic criteria for insulin resistance was hyperinsulinaemic-euglycaemic glucose clamp technique . But no obvious association between BD and insulin resistance was observed when the diagnostic criteria was the homeostasis model assessment of insulin resistance (HOMA-IR) formal score .Results for the analysis of the relationship between BD and insulin resistance are shown in Supplementary Fig. 6 , 27, 29.P = 0.02; Supplementary Fig. 8A) and dyslipidemia . Instead, the result indicated that there is no significant association between BD and diabetes mellitus .Besides potential immunologic interactions with metabolic processes, other factors including disease activity and treatment can also affect the risk of diabetes mellitus, arterial hypertension and dyslipidemia. In order to exclude the effect of drugs on the study, we excluded patients receiving drugs which may lead to metabolic complications such as steroids and DMARDs. The analyses showed similar results, and BD is significantly correlated with arterial hypertension . Furthermore, there was no significant heterogeneity or publication bias in the analysis. Further, the results of sensitivity analysis were stable and credible. Our findings revealed that BD is significantly associated with insulin resistance, diabetes mellitus and hypertension. Moreover, based on all the included studies, the association of BD with fasting blood glucose (a diabetes-associated parameter) and systolic blood pressure (a hypertension-associated parameter) was not statistically significant. However, there was an evident difference in DBP levels between patients with BD and non-BD controls; therefore, the DBP probability is a potential marker for predicting the incidence of BD-related hypertension. Still, the number of included studies reporting DBP levels was limited, and more studies are needed to confirm this result. In addition, when we excluded the effect of medications, the relationship between BD and diabetes was not statistically significant. Therefore, physicians should pay attention to the distinction between complications and drug effects in order to facilitate further treatment of patients.Consistent with previous reports, our study demonstrated a prominent association between BD and MetS. In the present study, the risk of MetS in patients with BD was 126% higher than in the healthy control group  are more predisposed to MetS than the general population (16.50%). An association between BD and the risk of MetS components—including diabetes mellitus, dyslipidemia, and hypertension—was also identified. Based on these findings, we recommend that physician consider these associations so that specific attention and treatment are available for patients with particular comorbidities. We also advise that patients with BD regularly monitor their blood pressure, triglyceride, fasting plasma glucose, and HDL cholesterol levels, as well as waist circumference.Supplementary file1 (DOCX 1252 KB)Below is the link to the electronic supplementary material.


Sinorhizobium meliloti 1021 bacteria secretes a considerable amount of flavins (FLs) and can form a nitrogen-fixing symbiosis with legumes. This strain is also associated with non-legume plants. However, its role in plant growth promotion (PGP) of non-legumes is not well understood. The present study evaluated the growth and development of lettuce (Lactuca sativa) and kale  plants inoculated with S. meliloti 1021 (FL+) and its mutant 1021ΔribBA, with a limited ability to secrete FLs (FL−). The results from this study indicated that inoculation with 1021 significantly (p < 0.05) increased the lengths and surface areas of the roots and hypocotyls of the seedlings compared to 1021ΔribBA. The kale and lettuce seedlings recorded 19% and 14% increases in total root length, respectively, following inoculation with 1021 compared to 1021ΔribBA. A greenhouse study showed that plant growth, photosynthetic rate, and yield were improved by 1021 inoculation. Moreover, chlorophylls a and b, and total carotenoids were more significantly (p < 0.05) increased in kale plants associated with 1021 than non-inoculated plants. In kale, total phenolics and flavonoids were significantly (p < 0.05) increased by 6% and 23%, respectively, and in lettuce, the increments were 102% and 57%, respectively, following 1021 inoculation. Overall, bacterial-derived FLs enhanced kale and lettuce plant growth, physiological indices, and yield. Future investigation will use proteomic approaches combined with plant physiological responses to better understand host-plant responses to bacteria-derived FLs. The challenges imposed by sustained climate change and global warming have had tremendous negative impacts on the global food supply chain and human health. Part of this challenge is caused by increased population growth and increased food and nutrition insecurity . To feedProviding sufficient food for an increasing human population will require different strategies and approaches. The usage of plant growth-promoting (PGP) microbes as biofertilizers or biocontrol agents is a promising alternative to support eco-friendly and sustainable agricultural production systems. PGP microbes can enhance plant growth and protect plants from biotic and abiotic stresses through the facilitation of nutrient uptake from the soil and atmosphere, synthesis of specific compounds, and assisting the plant to tolerate and adapt to changes in environmental factors . SeveralOryza sativa) [Zea mays) [Hordeum vulgare) and wheat (Triticum aestivum) [Lactuca sativa) [Capsicum annuum) [Solanum lycopersicum) [Helianthus annuus) [Rhizobia are well-known PGP bacteria, that can infect or colonize the roots of legumes to form root nodules and are best known as symbiotic nitrogen-fixing bacteria. However, many reports suggested that rhizobia could also influence and colonize roots of non-legumes, as they do with legumes ,26. Prev sativa) ,28, maizea mays) , barley estivum) ,31, lett sativa) , pepper  annuum) , tomato ersicum) , and sun annuus) , forms a annuus) . Biswas  annuus)  and Yann annuus)  pointed S. meliloti 1021 also enhances plant growth and development not only in leguminous crops but also in non-leguminous crops [Several studies indicated that us crops ,37. For us crops ,36,38,39ribBA, ribD, ribH, and ribE/C genes convert one molecule of guanosine triphosphate (GTP) and two molecules of ribulose5-phosphate into RF [ribBA gene [ribBA genes code for a putative bifunctional enzyme with dihydroxybutanone phosphate synthase and guanosine triphosphate (GTP) cyclohydrolase II activities, which catalyzes the initial steps of the RF biosynthesis pathway. It is believed that RibBA is a vital protein for enzymes responsible for catalyzing RF biosynthesis [Sinorhizobium meliloti strain 1021, the deletion of ribBA did not result in RF auxotrophy. The strain missing ribBA retained considerable GTP cyclohydrolase II activity but secreted a smaller amount of FLs, compared to the wild type, which ultimately affected bacterial root colonization [S. meliloti strains 1021 (FL+) and S. meliloti 1021ΔribBA (FL−) to test our hypothesis by using germination pouch assays and greenhouse potted-plant trials. The hypothesis of this study is that bacteria-derived FLs will improve the growth, physiology, yield, and phytochemicals in kale  and lettuce (Lactuca sativa).The genetic organization of RF biosynthesis genes varies among bacteria. In bacteria, the enzymes encoded by the  into RF . FurtherbBA gene . The ribynthesis . Howevernization . Based o+ inoculated seedlings (FL+ seedlings) compared to FL− inoculated seedlings (FL− seedlings). The inoculation with FL+ significantly (p < 0.05) increased the total root length, total root surface area, and total hypocotyl surface area of kale seedlings compared to FL− seedlings  increases in root and hypocotyl growth parameters, such as total length and total surface area (p < 0.05) increased by 13.7% and 14.3% in FL+ lettuce seedlings, respectively, compared to FL− lettuce seedlings. Total hypocotyl length and surface area were higher  in FL+ lettuce seedlings compared to FL− seedlings (ace area . The toteedlings .p > 0.05) different in the number of leaves, plant height, and stem diameter between FL+ inoculated plants (FL+ plants) and FL− inoculated plants (FL− plants). However, the plant height and stem diameter were higher in FL+ plants than in the FL− plants and control plants (+ plants than in FL− plants (+ plants compared to the FL− and control plants (p < 0.05) difference in the stem diameter of kale between the FL+ plants and no inoculation controls  between treatments  different between treatments (+ plants than in the FL− and control plants (p < 0.05) increased in FL+ plants compared to the control plants (p < 0.05) increased in the FL+ plants compared to the control plants (+ plants compared to the other treatments (Lettuce plants inoculated with FLeatments . The letl plants . The numl plants . Moreovel plants . The freeatments .a and b and carotenoid content in the leaves were significantly (p < 0.05) increased in bacteria-inoculated plants compared to control plants, although they were not significantly (p > 0.05) different between FL+ and FL− kale plants  higher in FL+ plants by 28% and 12% more than in the FL− and control plants, respectively (+ plants had significantly (p < 0.05) higher total phenolic content than the FL− and control plants  different between FL+ and FL− plants, chlorophyll a and chlorophyll b were higher in the FL+ plants compared to the FL− plants (p < 0.05) higher in the FL+ plants compared to the FL− and control plants  in the FL+ plants compared to the FL− lettuce plants. Among the other nutritional, calcium, sodium, manganese, and zinc were increased by 2%, 24%, 18%, and 3% in the FL+ plants, respectively, compared to the FL− plants  seedlings showed the highest root and hypocotyl growth compared to the S. meliloti 1021ΔribBA (FL− strain). Moreover, FL factors can both directly and indirectly enhance plant growth. For example, flavoproteins influenced Arabidopsis plant growth and development of hormonal levels, especially the metabolism of auxin and cytokinin [Many cellular processes in plants directly depend on FLs, as flavoenzymes catalyze a broad spectrum of different reactions ,45,46. Tytokinin .S. meliloti 1021 improved plant physiological processes, mainly photosynthesis, leading to the enhanced yield of the kale and lettuce plants. Chlorophyll fluorescence indices are broadly used to determine the photosynthesis performance of plants [S. meliloti 1021 inoculation, although they were not significantly different (p > 0.05) between treatments. Chlorophyll has a key role in photosynthesis and the overall productivity of crops [a and b  were higher in FL+ plants than in the FL− plants and control plants. Carotenoids are accessory photosynthetic pigments that cannot transfer sunlight energy directly to the photosynthetic pathway, however, must pass their absorbed energy to chlorophyll and function as a photoprotector under excessive light incidence [+ plants than in FL− plants. In addition to N, we found out that phosphorus (P) in lettuce was enhanced by FL+. In kale, potassium (K) was increased in FL+ plants compared to the other treatments. Both P and K are essential elements that determine plant growth and productivity and are involved in several physiological processes, such as the activation of numerous enzymes, and the regulation of the cation–anion balance [A significant number of flavoproteins also play key roles in the primary and secondary metabolism of carbon, nitrogen, or sulfur assimilation, amine metabolism as well as the plant’s response to abiotic stress ,48,49,50f plants . Therefoof crops ,53,54 anncidence ,56. Theyncidence ,58. The ncidence . Nitrogencidence ,61. Furtncidence . Previouncidence  through ncidence . The pre balance . These p balance .+ plants. We demonstrated that FL-secreting bacterial inoculation in plants can be associated with enhanced phytochemical content in both kale and lettuce plants, as previously reported for herbal tea by Fu [Phenolic compounds are inherent in plants and are known as potential agents for preventing and mitigating many oxidative stress-related diseases . Our resea by Fu . In addiea by Fu . These fThis study was conducted in the Compost and Biostimulant Laboratory, Department of Plant, Food and Environmental Sciences, Faculty of Agriculture, Dalhousie University, Canada.Sinorhizobium meliloti strains 1021 (FL+) [ribBA (FL−) [S. meliloti 1021 strains were grown at 28 °C in minimum mannitol ammonium (MMNH4) medium [21 (FL+)  and 1021BA (FL−)  were use) medium .Brassica oleracea var. acephala ‘Dwarf Green Curled’) and lettuce (Lactuca sativa ‘Simpson’) seeds were purchased from Halifax Seeds . S. meliloti strains  were grown for 5 days in MMNH4 medium plates. Cultures were scrubbed and suspended in sterile water to prepare bacterial solutions with an OD600 of 0.6. The lettuce and kale seeds were sterilized with 10% sodium hypochlorite for 10 min and washed thoroughly three times with sterile distilled water (ddH2O). The cleaned seeds were treated by soaking in solutions of FL+, FL−, and sterile ddH2O, which was the control treatment, for 90 min. The treated seeds were air-dried under a laminar flow for 30 min. Ten seeds per treatment were placed in a 16.5 × 18 cm CYG germination pouch  with six biological replicates, and saturated with 25 mL of the respective solution per seed treatment.Kale . The WinRHIZO™2000 software recognized scanned digital root and hypocotyl images and analyzed morphological traits, such as total lengths and total surface areas of root and hypocotyl ,72,73.All statistical analyses were performed using Minitab software version 20 . Data obtained with different treatments were subjected to a one-way analysis of variance (ANOVA). Post hoc means were compared using Fisher’s least significant difference (FSD) at α = 0.05.−1 illumination at a light intensity of 300 μmol m−2·s−1, and relative humidity of 70%. The seedlings were transplanted into 15 cm diameter plastic pots with a 10.5 cm depth. The mean day/night temperature cycle in the greenhouse during the experiment from transplanting to final harvest was 28 °C/20 °C with 71% relative humidity. Supplementary lighting was set to a 16 h day length cycle using a 600 W HS2000 high-pressure sodium lamp with NAH600.579 ballast . The growing medium was a general-purpose Pro-mix BX potting mix and transplanted seedlings were acclimatized for a week under greenhouse conditions before treatment application. Treatments were inoculation with FL+ strain, FL− strain, and a control (without inoculation). Inoculum was prepared as described in Pot experiments were carried out from March to June and September to December 2022 in the Department of Plant, Food, and Environmental Sciences greenhouse as two trials with seven biological replicates. Seeds selection and sterilization processes were the same as described in The number of green leaves for each plant was counted at 4 weeks after transplanting (AT). Plant height was measured from the apical meristem to the collar of the stem. The fourth leaf of each plant was tagged to estimate the plant growth rate by measuring the leaf elongation rate. Chlorophyll fluorescence indices were determined from three leaves per plant at 30 days AT using a chlorophyll fluorometer . Leaves were first dark adapted for 25 min before the initial fluorescence yield (Fo) was measured, after that the maximum chlorophyll fluorescence (Fm) emitted was measured during a saturating light pulse. Variable chlorophyll fluorescence (Fv) was calculated as Fv = Fm − Fo. Fluorometric parameters, including maximum quantum efficiency (Fv/Fm), and potential photosynthetic capacity (Fv/Fo), were determined at each saturating pulse. Fresh weights of leaf samples from each treatment were recorded at the final harvest, i.e., 35 days AT, using a portable tabletop balance . Harvested leaves were composed of leaf blades attached to petiole that had ≤5% blemishes or damage on the leaf surfaces. Those with blemishes or damage > 5% of the total leaf surface area were culled and not considered edible or marketable and were not included in the yield data. The leaves were dried in a 52100-10 Cole-Parmer mechanical convection oven dryer  at 65 °C to constant weights for 72 h, to determine leaf tissue micro and macro elements at the Nova Scotia Department of Agriculture Laboratory, Truro, NS, using inductively coupled plasma mass spectrometry .a and b, and carotenoid contents in kale and lettuce leaves were estimated as described by [g for 15 min and the absorbance of the supernatant was measured at 646.8, 663.2, and 470 nm against 80% acetone as a blank using a Bio-Tek Synergy H1 Hybrid Multi-Mode Reader and the Gen5 software application version 3.08 . Leaf chlorophylls a and b, and the total carotenoid contents of leaves were calculated by the following formulas and were expressed as a μg/g fresh weight (FW).Chlorophylls ribed by ,75, withg for 5 min before 100 μL of the supernatant was added to 200 μL of 10% (v/v) Folin–Ciocalteau reagent. The mixture was incubated in the dark at 25 °C for 2 h, after the addition of 800 μL of 700 mM sodium carbonate (Na2CO3). The absorbance of the supernatant was measured at 765 nm, using 95% (v/v) methanol as the blank. TPC was calculated using a gallic acid standard curve and expressed in terms of GAE (mg) per FW (g).Total phenolic content (TPC) was estimated using the Folin–Ciocalteu method, in terms of gallic acid equivalent (GAE) in mg/g of the extract, as reported by , with so3) colorimetric method, as described by Chang [g for 15 min. The reaction mixture containing 1.5 mL of 95% methanol, 0.1 mL of 10% AlCl3, 0.1 mL of 1 M potassium acetate, and 2.8 mL of distilled water was added to 500 μL of the supernatant. The solution was incubated at room temperature for 30 min and the absorbance was measured at 415 nm, using 95% (v/v) methanol as the blank without AlCl3. For TFC determination, quercetin was used to make the standard calibration curve and it was expressed in μg quercetin/g FW.Total flavonoid content (TFC) was calculated using the aluminum chloride . Data obtained with different treatments were subjected to a one-way analysis of variance (ANOVA). Post hoc means were compared using Fisher’s least significant difference (FSD) for plant growth, physiology, and yield parameter analysis and Tukey test for phytochemical analysis at α = 0.05. GraphPad Prism 9 software was used for visualization purposes.+ wildtype strain significantly increased the growth of kale and lettuce plants compared to inoculation with a FL− mutant strain, which secrete fewer FLs than S. meliloti 1021. The total phenolics and total flavonoid contents in the kale and lettuce leaves were significantly increased with bacterial FLs secretion. Two of the primary elements, nitrogen, and phosphorus, were increased by the bacterial-derived FLs in lettuce. Moreover, inoculation with FL+ apparently improved plant growth better than FL−. This study has the great potential to aid the development of a new methodology for improving the health and productivity of agriculturally important crops through inoculation of bacterial-derived FLs. However, in future investigations, we will use proteomic approaches combined with plant physiological responses to better understand host–plant responses to bacterial-derived FLs. Moreover, it will be important to investigate how bacterial-derived FLs impact on central carbon metabolism in kale and lettuce plants.This is the first study to examine the role of bacterial-derived FLs on plant growth and the development of green leafy vegetables. The results revealed that PGP from bacterial-derived FLs can enhance kale and lettuce growth and the development from the seedling to the harvest stage. The present results indicated that inoculation with a FL


Lactobacillus casei and Lactococcus lactis) were successfully engineered for scytovirin expression on the bacterial surface. These bacteria were found to be effective at neutralizing pseudotyped Ebolavirus in a cell-based assay. This approach can be utilized for prophylactic prevention, as well as for treatment. Since lactic acid bacteria can colonize the human body, a long-term efficacy could be achieved. Furthermore, this approach is also simple and cost-effective and can be easily applied in the regions of Ebola outbreaks in the developing countries.Scytovirin (SVN) is a lectin from cyanobacteria which has a strong inhibitory activity against Ebola virus infection. We engineered scytovirin as the inhibitor for surface display of lactic acid bacteria to block Ebola virus infection. Two different bacterial strains ( Ebolavirus in Zaire was first identified in 1976 in Africa (Ebola virus (EBOV) is an enveloped negative-stranded RNA virus which can cause severe Ebola viral disease (EVD) in humans and nonhuman primates . EBOV ben Africa , now fivn Africa : Zaire en Africa . The recn Africa . It is eScytonema varium (in vitro and in vivo (Scytovirin (SVN) is a small protein of 95 amino acids which was first identified from cyanobacteria of a varium . Scytovia varium . Becausea varium . Lectinsa varium . Due to a varium  and grifa varium , have bea varium  and havea varium . More st in vivo .in vitro data we have achieved successfully in this research direction.Because scytovirin demonstrated potent antiviral activity against Ebola infection, we used it to develop a novel live microbicide for Ebola infection control. This approach displayed scytovirin on the surface of Lactic acid bacteria (LAB) for delivery against Ebola virus infection. These bacteria can be delivered into mucosal surfaces of the body  that are the ports of viral entry where they can colonize and replicate. Previous studies using LAB expressing the antiviral lectin CV-N demonstrated that the bacteria can pass through the GI tract unharmed and stably produce lectins to inhibit HIV . Therefo22.1Escherichia coli DH5α and E. coli BL21 DE3 were used for cloning and initial protein verification, respectively. Both were cultured in LB media  overnight in a shaker at 250 rpm and 37°C. The antibiotics used for plasmid selection were 50 μg/mL kanamycin for pET28a and 250 μg/mL erythromycin for pLSVN3 and pLSVN7  at 37°C and 5% CO2 for 2–3 days until reaching log phase. For engineered bacterial culturing, 5 μg/mL erythromycin was added to the MRS media for applying selection pressure. HEK 293 T and TZM-bl mammalian cell lines were grown in Dulbecco Minimum Essential Media (DMEM)  supplemented with 10% FBS , 1 mM L-glutamine , and 100 μg/mL penicillin/streptomycin . Cells were cultured in a humidified cell culture incubator at 37°C with 5% CO2 using T-75 flasks.d pLSVN7 . L. case2.2SacI/XhoI restriction sites by GenScript and inserted into the vector pET28a  to test protein expression in E. coli BL21 (DE3) bacterial cells. SVN was overexpressed in E. coli previously by The scytovirin (SVN) gene sequence (GenBank: P86041.1)  with an 2.3Lactate dehydrogenase promoter (Pldh), signal peptide (SP), cell membrane anchor protein (ANC), E-tag marker (E), and the protein marker (GFP). The constructs were built up based on our previous plasmid pWZ486  by electroporation as previously described . Briefly2.5L. casei, the fusion protein was detected based on the GFP fluorescence. Bacteria were washed three times with PBS and analyzed on a BD FACSAria using a 488 nm laser. For the pLSVN7 construct transformed into L. lactis, the bacteria were first stained for 1 h with two primary antibodies: mouse monoclonal anti E-tag (Novus NBP2-67081) and rabbit polyclonal anti SVN . Goat anti mouse conjugated with AlexaFluor488  and goat anti rabbit conjugated with AlexaFluor594  were used as secondary antibodies, respectively. Bacteria were analyzed on a Beckman Coulter CytoFLEX FX at 488 nm and 561 nm. Unstained bacteria and stained wild-type bacteria were used in all experiments for an appropriate gating strategy.For the pLSVN3 construct transformed into 2.6Confocal microscopy was performed using the same fluorescent fusion protein or antibody combinations as described for flow cytometry (above). Following the antibody staining, bacteria were pelleted, resuspended in 10 μL PBS, and transferred to a microscope slide with cover slip. Images were captured using a Nikon A1R-Ti2  inverted confocal system.2.7ΔEnv (NIH HIV Reagent Program). The Ebola Envelope gene , was synthesized by GenScript and cloned into pcDNA3.1(+). Both plasmids were co-transfected into 293 T cells in a 10 cm plate using transfection reagent polyethyleneimine (PEI). Three days post transfection, the medium was harvested and centrifuged at 500 g to remove cell debris, and then the supernatants were stored at −80°C ] was added, and the samples were incubated at 37°C for 1 h in a heating block. Then, the samples were pipetted onto DEAE Filter mat circle papers (PerkinElmer), followed by three 10 min washes in 2X SSC buffer, and one 10 s wash in 100% ethanol. The filters were dried at room temperature and analyzed using a scintillation counter to measure the incorporation of 3H-dTTP into cDNA. The average CPM values from duplicates were determined.The titers of pseudotyped viruses were determined by Reverse transcriptase assay (RTA) . 500 μL 2.97/mL) in 1.5 mL micro-centrifuge tubes. The mixtures of bacteria and viruses were incubated for 1 h at room temperature. Then the tubes were spun for 1 min at a 13,000 g to remove the bacteria and bound pseudotyped virus. The supernatants were collected, and the viral titers determined by RTA.Pseudotyped Ebola virus stocks were mixed with wild-type bacteria or engineered bacteria  in a Veritas Luminometer.For the neutralization assay, pseudotyped Ebola virus was mixed with wild-type or engineered bacteria in the same manner as the adsorption assay described above. After centrifugation to remove the bacteria and bound pseudovirus, the remaining supernatants were applied to TZM-bl cells which were used as the target cells due to their ability to express luciferase when infected . The TZM2.11t-test at p-value ≤0.05.Statistical analyses were conducted for virus adsorption and virus neutralization data using GraphPad Prism software (version 9.0). The significances were determined by using unpaired two-tailed Student’s 33.1SacI/XhoI sites. The SVN plasmid was transformed to E. coli BL21 (DE3) cells for protein expression. The SVN fusion protein was observed under induction of 1 mM IPTG by Coomassie blue staining. The induced band of ~16kD fusion protein was noticed clearly  that appeared is the partial fragment of the fusion protein complex without the anchor domain (ANC). Flow cytometry analysis revealed the rate of SVN-GFP positive cells was 33.5% (The SVN-E-tag construct was cloned into our previous plasmid (pWZ486) through s pLSVN3 . This GF and 037 . To engiei cells . A ~ 75kei cells , suggestas 33.5%  a and b.as 33.5%  b. The bas 33.5% . The viras 33.5% . These m3.3Lactococcus lactis is another type of LAB which has a round shape and is widely used in the production of buttermilk and cheese. Thus, it is a very safe host strain candidate for anti-Ebola infection. We created the construct to remove the protein marker GFP but keep the E-tag marker from the construct pLSVN3, and this new construct was designated as pLSVN7  lysate as the positive control was also shown in the Western blots  and SVN (red), respectively. The results demonstrated that the SVN fusion protein is clearly expressed on the surface of the bacterium. The single color, green or red and the merged color of green and red indicated the overlapping presence of fusion protein surface expression  through non-specific binding, but the specific binding of the engineered bacteria to the viruses and are more than twice as effective, suggesting the SVN-expressing L. lactis bacteria can offer better protection against Ebola virus infection.Functional studies were performed against the pseudotyped Ebola viruses. First, we used the viral particles adsorption method to evaluate the bacterial ability for capturing the viruses. We then tested the engineered strain activity to inhibit Ebola virus infection using TZM-bl cells. The virus adsorption and neutralization data are shown in  control ,B. In co4L. lactis (92.4%) than in L. casei (33.5%), suggesting that lower positive rate in L. casei is due to the higher genetic instability because certain bacteria may lose the SVN plasmid or SVN gene during the replication, especially when the antibiotic pressure is reduced. Another possibility is the genetic recombination between the SVN plasmid and host genome by which the plasmid composition could be changed or damaged. We found that LAB were capable of quickly developing resistance to erythromycin (5 μg/mL) selective pressure, especially when grown in stationary, liquid culturing conditions, suggesting that the bacteria lose the plasmid (data not shown). Thus, increased erythromycin concentration is usually needed to get more genetically stable strains. In general, using a genomic integration method for engineering would be better than plasmid transformation for making genetically stable strains, which is essential for clinical or therapeutic use. In this research, L. lactis was found to be highly stable and would be assessed for in vivo (mice) colonization, and for protective efficacy against challenge with infectious Ebola viruses conducted in the BSL-4 containment.Bacterial therapy is a promising approach for human health and the common use of probiotics demonstrates its great potential . Using cin vivo applications by avoiding the potential for immunogenicity and cytotoxicity caused by the GFP protein  to determine the stability of the L. lactis strain under different physiological conditions, as well as determining the efficacy of the bacteria to prevent in vivo infection prior to advancing to clinical trials. In conclusion, the commensal bacterial based anti-Ebola approach is promising and will be beneficial for combating this deadly viral disease.In this report, our data from the nfection , and it The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/supplementary material.Ethical approval was not required for the studies on humans in accordance with the local legislation and institutional requirements because only commercially available established cell lines were used. Ethical approval was not required for the studies on animals in accordance with the local legislation and institutional requirements because only commercially available established cell lines were used.JW: Data curation, Methodology, Writing – review & editing, Formal analysis, Investigation, Validation, Visualization. NN: Data curation, Formal analysis, Investigation, Methodology, Validation, Writing – review & editing, Visualization. WW: Data curation, Formal Analysis, Investigation, Methodology, Writing – review & editing. LW: Formal Analysis, Investigation, Methodology, Writing – review & editing. HH: Investigation, Writing – review & editing. S-HX: Data curation, Investigation, Writing – review & editing, Conceptualization, Formal Analysis, Funding acquisition, Methodology, Project administration, Resources, Supervision, Writing – original draft.


Patient satisfaction with the results of their total knee arthroplasty (TKA) is one of the primary goals of this elective procedure. Furthermore, the association between the fulfilment of patients’ expectations and their satisfaction is well known. The aim of this study was to identify the key expectations of patients awaiting a TKA, evaluate their fulfilment, and compare the outcomes between very and not fully satisfied patients.A prospective cohort study of patients with knee OA scheduled for primary TKA was performed. Pre- and one-year postoperatively patient-reported outcome measures (PROMs) were assessed. Expectations and their fulfilment were evaluated via a questionnaire encompassing 31 expectations. Preoperatively, expectations were indicated as mandatory, desirable and not important. Postoperatively, fulfilment was rated as exceeded, fulfilled, partially or not fulfilled, and not applicable. Satisfaction with the results of TKA was measured with a numeric rating scale (NRS) of 0–10. Discrimination between not fully satisfied and very satisfied patients was set at ≥ 8, as has been proposed recently. To identify independent predictors of this discrimination, a multivariate logistic regression analysis was performed.Complete data sets of 352 patients were analysed. A set of 17 key expectations was identified. Relief of knee pain was fulfilled the most, and improvement of physical function was fulfilled the least. When asked about overall fulfilled expectations, 40% of patients rated them as exceeded, 34% as fulfilled and 26% as less fulfilled than expected. Not fully satisfied patients showed significantly lower PROMs pre- and postoperatively and less fulfilled key expectations. Higher numbers of exceeded and fulfilled mandatory expectations, higher overall fulfilment and better range of motion (ROM) were significant predictors for satisfaction ≥ 8.Patients’ expectations of TKA outcomes were high with equal emphasis on knee-related and general health-related aspects. Their fulfilment was positively associated with satisfaction. Surgeons should ask patients about mandatory expectations for successful TKA and counsel them about the likelihood of their fulfilment to avoid unrealistic expectations.II.The online version contains supplementary material available at 10.1007/s00167-022-07301-y. The decision for this surgery is associated with a number of individual outcome expectations , 34, 41.fter TKA .As largely acknowledged, a considerable number of patients remain not fully satisfied after TKA . ProportIt was hypothesised that patients present high expectations before surgery, but not all would be fulfilled postoperatively. Furthermore, it was hypothesised that greater fulfilment of expectations and better outcomes leads to very satisfied patients.This prospective cohort study has been performed in compliance with the Helsinki Declaration and has been approved by the ethics committee of the TU Dresden (EK 423112014).Between 09/2017 and 11/2019, all patients with knee OA scheduled for primary TKA surgery in a university hospital were informed about this study and asked to participate. Inclusion criteria were patients with advanced knee OA (grade 3 and 4 Kellgren and Lawrence), primary TKA , ability to understand German language, and signed informed consent. Patients were handed a set of PROMs including Oxford Knee Score , EuroQoLAll surgeries were performed by three different surgeons using a medial parapatellar approach without a tourniquet. All implants were cemented and no patellar resurfacing was performed. Full weight-bearing was allowed immediately and patients completed a standardised rehabilitation protocol.2 (SD 5.9), 54.5% were female, and 53.4% had serious comorbidities (ASA score 3 or 4). The majority of 328 patients (93.1%) received a bicondylar TKA and 24 patients (6.9%) needed a rotating-hinge prosthesis in severe valgus deformity.Between 09/2017 and 11/2019, altogether 441 patients received a primary TKA and 392 participated in this study. Until the one-year follow-up, six patients had died and one revision had occurred due to peri-prosthetic infection whilst undergoing oncological chemotherapy two months after surgery; 33 patients did not complete the follow-up, resulting in 352 complete data sets for analysis . Data are reported as mean and standard deviation (SD) for continuous values and absolute and relative frequencies for categorical values, respectively. Comparison between time points was performed by paired t test for continuous and by McNemar’s respective Wilcoxon signed-rank test for categorical data. Based on the proposed cut-off by Tolk et al. [t test for continuous and by chi-squared respective Mann–Whitney U test for categorical data. The significance level was set at p < 0.05.Sample size calculation was not conducted due to the exploratory design of this prospective study. All statistical analyses were performed using SPSSk et al. , the oveR2 were calculated [R2 has an upper bound of less than 1.0, whilst Nagelkerke R2 is an adjusted version of CoxSnell R2 and ranges between 0 and 1. The higher the R2 value, the better the fit between the model and the data.To determine key expectations, a threshold of 75% of patients indicating them as mandatory  was defined. Most important key expectations were identified by a threshold of 90%. This approach was recently introduced by a study investigating the expectations of patients before undergoing total hip arthroplasty (THA) . Expectalculated . CoxSnelWithin one year after surgery, 12 re-operations were performed . In addition, 11 patients required manipulations under anaesthesia.Out of 31 expectations, patients indicated a mean of 23 (SD 5.9) as mandatory  for a successful TKA and 5 (SD 4.7) as desirable . Six expectations were rated as mandatory by at least 90% of patients (most important key expectations) and another 11 by at least 75% of patients (key expectations)  is presented in Fig. Satisfaction with the results of surgery was rated a mean of 8.1 (SD 2.1), the distribution of answers on the satisfaction NRS 0–10 is shown in Fig. 247 patients indicated a satisfaction score of  ≥ 8 (70.2%), and 105 patients had a satisfaction score of < 8 (29.8%) on the NRS. Not fully satisfied patients were significantly older, had more comorbidities, had worse ROM postoperatively, and showed significantly lower PROMs (Table p < 0.001). Proportions of exceeded, fulfilled, partially or not fulfilled main and secondary goals were significantly different  in very satisfied and a mean of 8.0 (SD 3.2) in not fully satisfied patients , a higher overall fulfilment of expectations on the global rating scale (p = 0.038), a higher number of fulfilled main goals (p = 0.002), and better ROM postoperatively (p = 0.010). Table In multivariate logistic regression analysis, significant predictors for allocation into the group of very satisfied patients (NRS ≥ 8) were a higher number of exceeded main goals Below is the link to the electronic supplementary material.


The study aims to increase understanding of edaravone’s efficacy and safety as an amyotrophic lateral sclerosis (ALS) treatment and provide significant insights regarding this field’s future research.We conducted a comprehensive search of the Embase, PubMed, Cochrane Library, Web of Science, and Scopus databases for randomized controlled trials and observational studies up until September 2022. We evaluated the studies’ quality using the Cochrane risk of bias tool and the National Institutes of Health tool.P = 0.02), , and , respectively. However, the administration of edaravone did not result in any significant difference in adverse effects or efficacy outcomes between the two groups, as indicated by a P value greater than 0.05.We included 11 studies with 2845 ALS patients. We found that edaravone improved the survival rate at 18, 24, and 30 months (risk ratio (RR) = 1.03, 95% confidence interval (CI) [1.02 to 1.24], Edaravone improves survival rates of ALS patients at 18, 24, and 30 months with no adverse effects. However, edaravone does not affect functional outcomes. In order to ensure the validity of our findings and assess the results in accordance with the disease stage, it is essential to carry out additional prospective, rigorous, and high-quality clinical trials. The current study offers preliminary indications regarding the effectiveness and safety of edaravone. However, further comprehensive research is required to establish the generalizability and sustainability of the findings.The online version contains supplementary material available at 10.1007/s10072-023-06869-8. Amyotrophic lateral sclerosis (ALS) is a degenerative neurological condition characterized by the gradual deterioration of the motor neurons located in the brain and spinal cord, which are responsible for regulating voluntary muscle movement , 2. ALS Although the pathophysiological mechanisms and environmental factors that affect the disease are still unclear, the evidence suggests that free radicals play a crucial role in the progression of ALS , 9. FreeAlthough many drugs have been tested to treat ALS, riluzole was the only approved drug for about 20 years. Riluzole functions as an anti-glutamatergic agent by diminishing the discharge of glutamate, a neurotransmitter that is hypothesized to be involved in the deterioration of motor neurons in ALS. Clinical trials have demonstrated that riluzole has the potential to decelerate the advancement of ALS and extend the patient’s lifespan , 14. AddEdaravone is a newly tested free radical-scavenging medication that prevents neural cell damage by eliminating hydroxyl radicals and lipid peroxides , 16. ManThe authors conducted a systematic review and meta-analysis in adherence to the most recent PRISMA updates and Cochrane standards , 25.We searched PubMed, Cochrane library, Embase, Scopus, and Web of Science databases for relevant trials until September 2022. Our search was done using the following search strategy: (edaravone OR norantipyrine OR norphenazone OR edarabone OR radicava OR fraseda OR radicut OR phenylmethylpyrazolone OR methylphenylpyrazolone OR nuravon OR aravon OR “MCI 186” OR “MCI-186” OR “MCI186”) AND .We included randomized control trials (RCTs) and observational studies that compared edaravone versus control in patients with ALS regarding safety, efficacy, and survival outcomes. We removed the duplicates using EndNote software and screened the title and abstract of all the remaining studies. Studies that might fit the eligibility criteria were full-text screened. Additionally, we screened the references of the included articles to find any missing relevant studies. Two authors did the previous steps, and the third author solved any conflict.The Cochrane risk of bias tool (version 1) was used to assess the degree to which the studies under consideration may have been biased . The purSummary data: study design, site, registration, follow-up duration, inclusion criteria, and outcomes.Baseline data: arms, patients’ number, age, gender, ALS diagnosis definite ALS diagnosis probable, ALS severity grade, duration of disease, riluzole use, and ALS Functional Rating Scale-Revised (ALSFRS-R score) before the observation period.Outcomes:Primary outcomes include survival rates, changes in the 40-item ALS assessment questionnaire (ALSAQ-40 score) and ALSFRS-R score, and any adverse events or serious adverse events.Secondary outcomes: changes in forced vital capacity (FVC), grip strength in Kg and modified Norris scale score, and other adverse events .The following data were retrieved in Excel sheets:P value < 0.05. We assessed the heterogeneity using I-square (I2) and chi-square tests. Homogeneous data (P ≥ 0.1 or I2 < 50%) were pooled in a fixed-effect model, while heterogeneous data (P < 0.1 or I2 > 50%) were analyzed in a random-effect model. We performed subgroup analyses depending on the number of cycles and months of survival.The process of analyzing data was carried out through the utilization of Review Manager (RevMan) software version 5.4. The measure of effect utilized in the study was either risk ratio (RR) or mean difference (MD), along with 95% confidence intervals (CI), based on the nature of the data (dichotomous or continuous). We reported the significance at After removing duplicate results, the initial search returned a total of 720 unique articles. A subset of 23 studies were determined to be suitable for full-text assessment after undergoing a thorough screening process that involved evaluating the relevance of titles and abstracts. In the end, eleven articles fulfilled the inclusion criteria for our quantitative study after a thorough evaluation , 28–33 .Fig. 1PR).The studies included in this review comprised four RCTs –21 and s2. Our cohort studies had a fair quality; the details are in Online Resource (Online Resource 2).The quality of the RCTs that were incorporated in our analysis was rigorously evaluated to ensure the dependability and accuracy of our results. The majority of the RCTs exhibited a low probability of bias, suggesting a considerable degree of methodological quality and a reliable basis for our analysis. Upon further examination, it was discovered that the study conducted by Eishi-Oskouei et al. exhibited a higher probability of bias , which rFig. . Our cohP = 0.0004). The results obtained through pooling of data were found to be heterogeneous   Fig. .Fig. 3A P value = 0.45) and heterogeneous   Fig. .P = 0.23). The results obtained through pooling of data were found to be heterogeneous  . The results obtained through pooling of data were found to be heterogeneous  . The results obtained through pooling of data were found to be heterogeneous   Fig. .P = 0.03). The results obtained through pooling of data were found to be homogeneous  . The results obtained through pooling of data were found to be homogeneous   showed no difference between both groups . The results obtained through pooling of data were found to be heterogeneous  . The results were homogeneous  . The results obtained through pooling of data were found to be heterogeneous  . The results obtained through pooling of data were found to be heterogeneous  . The studies were heterogeneous  . The results obtained through pooling of data were found to be homogeneous  . The results obtained through pooling of data were found to be homogeneous   revealed insignificant variation between both groups . The results obtained through pooling of data were found to be homogeneous  . Heterogeneity was observed  .Five articles reported the FVC for 782 ALS patients. There was no improvement in FVC score in ALS patients treated with edaravone . The results obtained through pooling of data were found to be homogeneous  .The analyzed data (sample size = 350) showed no variation between the edaravone and control groups . The analysis showed homogeneity  . The data were homogeneous  .The pooled data of 358 ALS patients revealed insignificant differences . Our results were homogeneous  . Other adverse events such as gait disturbance, dysphagia, constipation, insomnia, and upper respiratory tract inflammation showed insignificant results (P > 0.05). Moreover, the most commonly reported adverse events were gait disturbance [.The results showed that the edaravone-treated patients had significantly lower musculoskeletal disorders than the control group  and Lunetta et al. (2020) [P = 0.53), respectively. The results remained significant (P < 0.05) . The heterogeneity of the change in ALSFRS-R score after six cycles was solved by removing Okada et al. (2018) [P = 0.22). The results remained insignificant (P = 0.3) . The results of FVC were homogeneous (P = 0.77) and still insignificant (P = 0.12) after sensitivity analysis .The survival rates at 18 and 24 months were homogeneous after excluding Houzen et al. (2021)  (P = 0.1. (2020)  (P = 0.5. (2018)  Supplementary file2 (DOCX 19 KB)Supplementary file3 (PDF 256 KB)Below is the link to the electronic supplementary material.


A comorbidity of anorexia nervosa (AN) and myalgic encephalomyelitis (ME/CSF) is uncommon. A 17 years-old male adolescent with possible onset of ME/CFS after an Epstein Barr Virus infection (EBV) and later onset of AN during a second period of weight loss was twice treated off-label with metreleptin for 15 and 11 days, respectively. As in previous cases, eating disorder specific cognitions and mood improved. Interestingly, fatigue and post-exertional muscle pain (P-EMP) improved, too. We discuss potential mechanisms. Treatment with metreleptin may prove beneficial in AN and in ME/CSF associated with substantial weight loss. Metrele changes , 7, 8.Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CSF) is one of the post-acute infection syndromes . An infeFatigue, post-exertional malaise, neurocognitive problems, and unrefreshing sleep are common in ME/CSF . AdditioWe describe a currently 18 years old male who had developed both probable ME/CSF and AN after an Epstein Barr virus (EBV) infection. Family history was uneventful apart from a major depressive disorder (MDD) of the maternal grandfather. The patient had discrete separation anxiety upon entrance to kindergarten and speech therapy for dyslalia. During childhood, otitis media was treated repeatedly with antibiotics, a salmonella infection required a 5 days hospitalization. The achievement-oriented youth played a ball game competitively at the national level. After otherwise normal somatic, emotional, social and cognitive/academic development the patient fell ill at age 15 years and 8 months with an EBV infection  with splenomegaly and enlarged lymph nodes. His physical condition deteriorated entailing repeated brief hospitalizations. Weakness, loss of appetite and constipation were prominent.After exclusion of systemic infections, hemato-oncologic disorders (bone marrow biopsy), autoimmune, endocrine and neuromuscular disorders  CFS was diagnosed. The patient was intermittently able to walk a few steps only with physical exertion induced muscle pain in the upper arms and legs and post-exertional malaise which could persist the following day. Weakness and tingling paresthesia as well as episodes of headaches and dizziness were noted 3 months after onset of the infection. After 6 months nerve pain appeared at the right neck triangle and celiac disease (Marsh type 3b) was diagnosed via biopsy.The introduction of a gluten free diet led to an improvement of constipation; a second biopsy conducted 5 months later revealed normal results. Weight loss amounted to 15 kg 7 months after onset of EBV infection , after which a body weight of around 53–55 kg was maintained for approximately 12 months .2; He was admitted for inpatient adolescent psychiatric treatment at age 17 years and 8 months his body weight had dropped to 42.6 kg  were used to track emotions and cognitions daily ; the immThe patient was concomitantly treated with olanzapine 7.5 mg (3 × 2.5 mg) daily. Psychoeducation focused on expected treatment effects, the presumed underlying mechanism , and the necessity to use the boost in motivation to gain weight in order to enable sufficient endogenous leptin secretion.Metreleptin (3 mg/days) was applied subcutaneously at 9:00 am for 11 consecutive days. Again, the patient was concomitantly treated with 3 × 2.5 mg olanzapine daily. Psychotherapeutic support was provided throughout the hospital stay, including both dosing periods.d + 18 scores of the items feeling fat and fear of weight gain had rebounded completely.Similar to previous findings , daily mThe baseline responses provided in the CFSQ were compatible with a diagnosis of CFS. VAS items fatigue and muscle pain (evening ratings only) dropped rapidly after initiation of dosing . Sleep qd − 3) to 25 (day 4) and 20 (d + 3) to reach the low nadir of 16 on d + 13 (d + 18: 23). The raw score of the EDI-2 dropped from 377  to 331 .Self-rated depression BDI-II score dropped from 42 (2) exceeded the 99th percentile of leptin levels adjusted for sex, Tanner stage and BMI.During the initial 11 weeks of inpatient treatment serum leptin levels proved undetectable see . The levd + 79, muscle pain continued to be diminished. The patient reported no pain upon normal walking or marching. However, upon jogging muscle pain rapidly set in. Fatigue, too, was not viewed as a complaint any longer. Concentration problems persisted; the patient expressed frustration due to memory problems in school (attendance of 2 h daily only). He had reinitiated social contacts. Mood swings represented an additional complaint. Overall, his condition, including eating disorder related symptoms had further improved in comparison to discharge from inpatient treatment (d + 2). BMI increased over the course of the treatment  to 8 (day 9). BMI dropped from 18.8 kg/m2 by 0.4 kg/m2 during the dosing period but returned to the same value (18.8 kg/m2) at the end of the treatment period  he is working full-time as an apprentice in a bakery; eating disorder specific cognitions persist at a subclinical level and muscle pain was no longer reported. The current BMI is 19.11 kg/m2. He recently began sports activities again. A relapse did not occur, but cannot be excluded in light of the low BMI.After discharge from inpatient treatment the patient continued to struggle with AN related symptoms, which slowly improved. The BMI development over the whole course of the treatment is shown in This case report is remarkable for the rapid improvement of symptoms of both AN and probable ME/CSF upon two short-term metreleptin treatments. The positive changes set in shortly after the beginnings of both dosing episodes, but partly faded after the end these episodes, which is also reflected in the high baseline values of some VAS items at the beginning of the second dosing. We cautiously refer to probable ME/CSF, because we focused our clinical assessment on P-EMP; post-exertional malaise was not systematically assessed. We also did not evaluate the ME/CSF symptomatology with questionnaires based on the Canadian Consensus Criteria or ICC criteria , 22. Fin2. At this time point, weight phobia, body image disturbances, fatigue and evening muscle pain dominated the clinical symptomatology, which in combination with the severe underweight led to the DSM-5 diagnosis of AN in addition to preexisting probable ME/CSF.In our patient an Epstein–Barr virus infection initially resulted in weight loss, fatigue and P-EMP and malaise, for which no underlying medical condition was detected. Concentration and memory problems had plagued the patient from the beginning on. The initial weight loss of approximately 15 kg was substantial and plateaued for 12 months, after which further 11.5 kg were lost resulting in an exceedingly low referral BMI of 12.45 kg/mWeight loss can occur with EBV infection  and ME/CTriggered by a remark from one of our reviewers, we briefly discuss a potential immune system involvement as an overlapping pathophysiological hypothesis for the simultaneous occurrence of probable ME/CFS and AN. Immune dysregulation in ME/CFS is common and a subset of patients with ME/CFS may have an autoimmune etiology . Some stApart from fatigue, early onset P-EMP was pronounced in our patient; post-exertional malaise persisted until the next day. Within weeks after onset of the EBV infection, the patient complained of up to five nightly bladder voidings consistent with a genitourinary symptom of ME/CSF . In accoUpon hospitalization for treatment of AN 2.0 years after onset of EBV-infection, the longstanding P-EMP interfered with the urge to move, which is a common symptom of AN, as is hyperactivity , 35. WhiDuring the initial dosing period the patient received 3 to 6 mg per day. The parents had noticed strong changes during the dosing days with 6 mg. It is unclear if more pronounced improvements would have been associated with continuous application of 6 mg and/or a longer dosing period. Mood improvement (VAS) in the current case was somewhat less pronounced than in other patients; nevertheless, BDI-II scores were approximately halved after the dosing period. For an intermittent period of time mood improved around noon. Mood swings were rated high throughout dosing. Notably, these changes occurred despite an exceedingly high baseline serum leptin level adjusted for sex, Tanner stage and BMI, which likely reflects prior weight gain .The improvements in sleep quality, depression, and eating disorder cognitions have been reported previously in five patients with AN . In addiLeptin is assumed to contribute to the inflammatory state found in obesity that predisposes to metabolic syndrome, type 2 diabetes mellitus, and cardiovascular disease. However, leptin is also an important mediator of the immunosuppressive state in undernutrition .ob/ob mice  and acid-sensing ion channels , and minor(s)’ legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article. Written informed consent was obtained from the participant/patient(s) for the publication of this case report.JH: Conceptualization, Writing – original draft, Writing – review & editing. JA: Writing – review & editing, Visualization. LP: Data curation, Investigation, Writing – review & editing. CK: Data curation, Investigation, Writing – review & editing. BS: Data curation, Investigation, Writing – review & editing. GG-D: Data curation, Investigation, Writing – review & editing.


C. elegans.Prolonged fasting, characterized by restricting caloric intake for 24 h or more, has garnered attention as a nutritional approach to improve lifespan and support healthy aging. Previous research from our group showed that a single bout of 36-h water-only fasting in humans resulted in a distinct metabolomic signature in plasma and increased levels of bioactive metabolites, which improved macrophage function and lifespan in This secondary outcome analysis aimed to investigate changes in the plasma lipidome associated with prolonged fasting and explore any potential links with markers of cardiometabolic health and aging.2) in four metabolic states: (1) overnight fasted (baseline), (2) 2-h postprandial fed state (fed), (3) 36-h fasted state (fasted), and (4) 2-h postprandial refed state 12 h after the 36-h fast (refed). Plasma lipidomic profiles were analyzed using liquid chromatography and electrospray ionization mass spectrometry.We conducted a controlled pilot study with 20 male and female participants . After correction for multiple testing, 245 out of 832 lipid species were significantly altered in the fasted state compared to baseline (P < 0.05). Random forest models revealed that several lipid species, such as LPE(18:1), LPC(18:2), and FFA(20:1) were important features in discriminating the fasted state from both the overnight fasted and postprandial state.Several lipid classes, including lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylethanolamine, and triacylglycerol were significantly reduced in the 36-h fasted state, while free fatty acids, ceramides, and sphingomyelin were significantly increased compared to overnight fast and fed states (Our findings indicate that prolonged fasting vastly remodels the plasma lipidome and markedly alters the concentrations of several lipid species, which may be sensitive biomarkers of prolonged fasting. These changes in lipid metabolism during prolonged fasting have important implications for the management of cardiometabolic health and healthy aging, and warrant further exploration and validation in larger cohorts and different population groups. FastPF exhibits both beneficial and detrimental effects on human metabolism. One mechanism by which PF can beneficially affect health, and in fact has even been found to extend lifespan, is the reduction of oxidative damage, which is thought to be a normal part of the aging process . StudiesThe effects of PF on the plasma lipidome have gained attention as an area of interest. During fasting periods of 12 to 24 h, there is a metabolic shift from using glucose to utilizing lipid-derived ketone bodies and free fatty acids (FFA) as an energy source in response to caloric deprivation to maintain vital brain and tissue function . LipolysC. elegans  to measure four distinct nutritional states: (1) overnight fasted (baseline), (2) 2-h postprandial (fed), (3) 36-h fasted (fasted), and (4) an additional 2-h postprandial 12-h after the 36-h fasting period (refed) (The complete study design is described in detail elsewhere . Plasma  (refed) . Partici2.2.Blood samples from all participants were collected using ethylenediaminetetraacetic acid plasma tubes and subjected to centrifugation  for 10 min at 4°C. Following centrifugation, the samples were partitioned into smaller aliquots. Samples were preserved at −80°C until analysis.2.3.Lipidomic analysis was performed by Metabolon, Inc using their Complex Lipid Targeted Panel . Briefly, lipids were extracted from plasma in the presence of deuterated internal standards using the butanol:methanol method, as previously described . The ana2.4.post hoc analysis was performed using the function pairwise_t_test from the R package rstatix  models with cross-validation (number of partitions = 5) were used as an unbiased supervised machine learning approach for classification using the Tidymodels package . This meSaturation level of FA was determined by categorizing each lipid class based on the number of double bonds present in their respective FA. FA were classified as saturated, monounsaturated, or polyunsaturated depending on whether they had 0, 1, or 2 or more double bonds, respectively. FA species that were not present in one of the participants at any time point were excluded from the analysis. Total FA concentration for each lipid class was calculated by summing the concentrations of FA within each saturation level. The data was then log-transformed for linear modeling using the R package limma  to calcu3.3.1.P < 0.001 for all, P < 0.05), fed (P < 0.01), and refed state (P < 0.05) (P < 0.001) and fed state (P < 0.01) and remained significantly elevated in the refed state compared to baseline (P < 0.01) and the fed state (P < 0.05) . PE was significantly decreased in the fasted state compared to the baseline (P < 0.0001), fed (P < 0.0001), and refed states , while LCER remained significantly elevated in the refed state compared to both the baseline (P < 0.001) and fed states (P < 0.001). MAG was significantly increased in the fed state compared to baseline  . SM was  < 0.05) . PC [F . PCA reved state . Among ted state . Of the ed state . All 34 ed state . Out of Analysis of the 518 TAG species, spanning a range of 36 to 60 carbon atoms and 0 to 12 double bonds, revealed distinct patterns of change . TAG spe3.3.P < 0.0001 for all, Random forest analysis was used as a multiclass classification model to differentiate between the four nutritional states . The mod3.4.P-value < 0.05. In the comparison of the 36-h fasted state vs. baseline, MUFA LPE and PI were significantly reduced (P < 0.001 for both), as well as polyunsaturated acids (PUFA) LPE and LPC  and LPC(18:2) during the 36-h fasted state compared to both the overnight fasted and fed states. This finding is consistent with a study by Steinhauser et al., which demonstrated that several plasma LPC and LPE lipid species, including LPE(18:1) and LPC(18:2), were decreased during periods of starvation lasting between 1 and 10 days compared to an 8-h overnight fast . FurtherSphingolipids, including SM, CER, and their analogs DCER, HCER, and LCER are a family of lipids involved in cell recognition and signal transduction , 52. TheWe found that total PI concentration was not significantly different across the four nutritional states, while total MUFA PI species significantly decreased in the 36-h fast compared to the overnight fast. PI serves as a signaling molecule in the cell membrane involved in insulin signaling and glucose uptake . FastingA major strength of this study is the targeted quantitative analysis of the plasma lipidome, offering a precise and dependable evaluation of overall lipid alterations during PF. Furthermore, the same participants were measured at four different time points, which strengthens the internal validity of the study by reducing inter-individual variability. However, certain limitations should be considered. The small sample size limits the generalizability of our findings, necessitating further studies with larger, more diverse cohorts for validation. Additionally, the participants were healthy young adults recruited from the Davis, CA area, which may not fully represent the general population, further emphasizing the need for broader, more diverse samples in future investigations. Lastly, while our study reveals complex lipidomic changes during acute PF in a cohort of healthy young individuals, further research is needed to explore the underlying mechanisms of PF and their health implications more thoroughly, particularly in older or diseased populations. Future research could explore the long-term effects of repeated PF cycles on lipid metabolism and assess the potential benefits for individuals at risk of cardiometabolic diseases. Investigating the connection between PF and the sustained decrease in TAG, PE, and lysophospholipid levels with overall health outcomes would be particularly beneficial for individuals with metabolic or dyslipidemic disorders.In summary, we used targeted quantitative analysis to investigate the acute effects of PF on the plasma lipidome. Our findings revealed marked changes in various lipid classes, such as FFA, TAG, LPC, LPE, PE, and sphingolipids during a 36-h fast. These alterations in lipid metabolism could be attributed to the body's adaptation to the fasted state, with potential implications for cardiometabolic health and aging. This secondary outcome analysis contributes valuable insights into the complex lipidomic changes occurring during PF and establishes a foundation for future research to elucidate the underlying mechanisms and health implications of these lipid alterations.