Solanum tuberosum is one of the most important crops in the world; however, drought has caused significant losses in its production. One solution is the use of arbuscular mycorrhizal fungi (AMF). In this study, the phenolic profiles and antioxidant activity of the leaves of two potato genotypes (VR808 and CB2011-104) were evaluated over time in crops inoculated with two strains of AMF, as well as a consortium, in combination with a commercial fungicide. In addition, three usable humidity levels were established after the beginning of tuberization. The phenolic compounds found during the first sampling time in the VR808 genotype reached a maximum of 3348 mg kg−1, and in the CB2011-104 genotype, they reached a maximum of 2982 mg kg−1. Seven phenolic compounds were detected in the VR808 genotype, and eleven were detected in the CB2011-104 genotype, reaching the highest concentration at the last sampling time. The antioxidant activity in the first sampling was greater than the Trolox equivalent antioxidant capacity (TEAC), and in the third sampling, the cupric reducing antioxidant capacity (CUPRAC) predominated. The association of AMF with the plant by PCA demonstrated that these fungi assist in protecting the plants against water stress, since in the last harvest, the results were favorable with both mycorrhizae. Currentl impact  and is il impact . Global l impact . The potl impact ; it has l impact .Plants are constantly exposed to extreme conditions that cause stress, which affects their growth, development, and production. These conditions include abiotic factors, such as deficiency, or overabundance conditions such as drought, temperature, salinity, light, nutrients, flooding, and phytotoxic compounds . Stress Rhizophagus irregularis, two PIP aquaporin genes  were upregulated under drought stress, demonstrating a better performance of AMF root cells in water transport under water deficit, which is connected to the shoot physiological performance in terms of photosynthetic capacity [Robinia pseudoacacia L., the height increased by 100% with the inoculation of AMF with respect to the non-inoculated ones [Pistacia vera L. treated with AMF was greater than that of the treatment without AMF. The same was true for the contents of N and Ca [Arbuscular mycorrhizal fungi (AMF) are soil microorganisms that have existed for more than 400 million years and are morphologically unchanged . Mycorrhcapacity . Chitarrcapacity  have alscapacity  revealedted ones , and theN and Ca .Solanum lycopersicum plants increases between 50% and 83% compared to the control [Eriocephalus africanus plants have been reported [Rhizophagus fasciculatus, Funneliformis mosseae, and R. irregularis and exposed to water stress, their concentrations of phenols and flavonoids increased, which is related to greater antioxidant properties. This increase showed that mycorrhizae act as moderators and cause the production of secondary metabolites [Triticum aestivum was inoculated with two AMF strains, also increasing the total phenol content by 55% and 40%, compared to the control [G. intraradices) compared to noninoculated plants [Solanum tuberosum plants exposed to water stress have improved physiological and metabolic activities when inoculated with AMF, and the main aim of this study is to evaluate the effect of AMF inoculation on Solanum tuberosum crops exposed to water stress by evaluating the physiological and metabolic parameters of the inoculated and noninoculated plants.Plants synthesize primary metabolites for essential functions, such as growth and development, and secondary metabolites for more specific functions under drought conditions and before other types of environmental stresses, in which secondary metabolites play an important role in the adaptation and survival of stressed plants . There a control . In whea control . On the reported . The increported . Phenolsreported . In the abolites . Similar control . Potato d plants . AccordiDifferent profiles of flavonols and HCADs were detected in the different genotypes, and three harvests were performed during growth; however, in both cases, the HCADs corresponded primarily to caffeoylquinic acid isomers and the flavonols corresponded to quercetin and kaempferol derivatives . Quantit−1 and 6.5 to 22.5 mg kg−1, respectively, and the maximum value in both was from the MIX treatment  and CC treatments with S1 stress, with increases of 380% and 1560%, respectively, as compared to the control with normal irrigation , FRAP (ferric reducing antioxidant power) assay, TEAC , and DPPH  method are widely used .−1 and 8.7 and 11.6 µmol g−1, respectively, in which the highest values were detected in the CC and HMC26 treatments; however, in the TEAC, there were no significant differences between the treatments. In the CB2011-104 genotype . In addition, in all the antioxidant activity methods, a tendency to decrease with S1 and S2 stresses was observed; however, primarily in S1, there were no significant differences compared to the control with normal irrigation.During the second harvest, the antioxidant activity levels in the VR808 genotype D–G in TE−1 in the HMC26 treatment with a stress level of S1, but there were no significant differences between the treatments for this increase. TEAC, DPPH, and FRAP maintained their concentrations in ranges similar to the second harvest, showing a tendency to decrease with increasing stress. In genotype CB2011-104  was performed considering all the evaluated parameters. In the VR808 genotype of the first harvest A,B, the However, during the first harvest of the CB2011-104 genotype A,B, the C. lamellosum/REFLECTXTRA® for VR808 genotype, F. mosseae/MONCUT for CB2011-509 genotype, and C. lamellosum/MONCUT for CB2011-104 genotype [C. lamellosum and REFLECTXTRA® was the ideal combination in all genotypes [Solanum lycopersicum with differentiated irrigation, in which the concentrations of total phenols over time also showed a significant decrease of 83% [Solanum tuberosum genotypes [−1, and between 3891 and 3753 mg kg−1 in leaves, have been reported, which is related to our results, since in the first harvest of the VR808 genotype, a maximum of 3348 mg kg−1 was obtained, and in the CB2011-104 genotype, a maximum of 2982 mg kg−1 was obtained [Scabiosa maritima during development can be explained by the biochemical changes that occur during growth, which are related to the plant’s defenses against biotic and abiotic stresses [C. claroideum, showed a tendency to increase the concentrations of phenolic compounds, which is one of the important parameters in the tuber stage, regardless of drought stress during the first harvest. These levels were similar. The reason why these levels were lower in the following harvests is because this change occurred when the tuberization process began, which is explained by the high concentration of these compounds in the tuber analysis [Previous studies in other substrates, such as peat, reported higher significant differences between the various evaluated parameters. In addition, different profiles and higher concentrations of phenolic compounds have been detected in potatoes depending on the genotype, AMF, or fungicide . Subsequgenotype . On the enotypes . Notablyenotypes . The dece of 83% . This deenotypes . It is iobtained ,25. Intestresses . Water danalysis ,25. Phenanalysis . In wheaanalysis ,40,41. Tanalysis . The maianalysis .C. claroideum and HMC26, and similar antioxidant activity levels were observed in the first harvest [F. ananassa crops inoculated with C. claroideum also increased [Sesamum indicum L., the effects of AMF inoculation (F. mosseae with R. irregularis) on the biomass and grain yield improved under severe water stress and were more effective than optimal irrigation conditions. This finding revealed that inoculating sesame plants with mycorrhizal fungi improved the state of water retention, and the plant could modulate and neutralize severe water stress; in addition, fewer antioxidant enzymes are required to eliminate radicals and active oxygen [Zea mays L. under AMF inoculation, there is a lower concentration of MDA than in uninoculated plants [T. aestivum L. inoculated with F. mosseae under water stress produced improvements in the composition of phenols and antioxidant activities in the grains of the wheat plants, with an increase of up to 30.2% in the concentration of apigenin-C-pentoside-C-hexoside III under water stress with respect to normal irrigation [The antioxidant activity results are consistent with those reported by Fritz et al. , in whic harvest . Notablyncreased . Most AMncreased . Howeverncreased . Potato ncreased . In Sesae oxygen . In Zea d plants . Anotherrigation . Antioxirigation , the mosrigation .F. ananassa inoculated with C. claroideum, where the antioxidant activity by this method also had an increase [The CUPRAC method detects the antioxidant capacities of phenolic acids, flavonoids, carotenoids, and other polyphenols . Potato increase .Solanum tuberosum plants over time. In both potato genotypes, under different levels of usable moisture, important variations in the leaves in the amounts of compounds and antioxidant activities were observed, since the VR808 genotype initially had greater activity when unstressed conditions were observed, but ultimately, its higher expression was at the S1 stress level under inoculation with each mycorrhiza separately. In the CB2011-104 genotype, in the first harvest, the treatment with S1 is highlighted. However, in the third harvest, all the mycorrhizal strains helped to maintain their antioxidant activities at the S2 stress level. The results reveal that the AMF helps protect the plant against hydric stress, since the PCA results show related symptoms; however, they help protect the CB2011-104 genotype to a greater extent because it was the one that possessed a greater protection under the S2 stress level in the last harvest.Highlighted here are the various most relevant results obtained, which were based on the monitoring of phenolic and antioxidant responses in the leaves of Water, methanol, acetonitrile (HPLC grade), Folin-Ciocalteu reagent, ethanol, and formic acid were obtained from Merck . Phosphoric acid, TROLOX , ABTS ), DPPH , neocuproin (≥98%), quercetin (>95%), copper (II and III) chloride (grade pa), potassium persulfate, chlorogenic acid (≥95%), iron (III) chloride, calcium chloride, ammonium acetate, and methanol were obtained from Sigma-Aldrich . Sodium carbonate and TPTZ  were obtained from Supelco .Solanum tuberosum plants were grown in pots subjected to water stress conditions in the greenhouse of the Department of Chemical Sciences and Natural Resources of the Universidad de la Frontera, Temuco, where they were kept under 50% artificial shade by placing a mesh over the greenhouse, with a light/dark photoperiod of 16/8 h and day/night temperatures of 25/18 °C. Tubers of two genotypes were used (VR808: white potato and CB2011-104: purple potato), with different skin and pulp colors, which were provided by the company Papas Arcoiris Ltd.a. . At sowing time, the potato tubers were inoculated with two strains of AMF according to each treatment ; moreover, control treatments without mycorrhizal (WM) were included. Plants inoculated with one inoculum also received the filtrate of the other inoculum. The inoculation method was carried out according to the one described previously by Fritz et al. (2022) [®, as an agronomic management, was added in all treatments. The stress factor was applied under three conditions: 40% (S2), 70% (S1), and 100% (0) available soil moisture levels. After sowing, all plants were treated with normal watering until the beginning of tuberization. Watering was managed considering three usable humidities ; moreover, the first collection time was carried out. Soil and sand were used as substrates in a 2:1 ratio, and this substrate underwent a sterilization process and was autoclaved for 20 min at 121 °C and 1 atm pressure. Each treatment was applied in triplicate . The potato leaf genotypes were harvested for a period of three months, during which the first harvest was 60 days after sowing (DAS), the second was 90 DAS, and finally the third was 120 DAS, and the leaves were immediately stored at −80 °C until analysis.. (2022) . The AM 18 column  and a Novapak Waters C18 guard column  at 40 °C and a flow rate of 0.55 mL min−1. The mobile phases were 92:3:5 (v:v:v) water, acetonitrile, and formic acid (mobile phase A), and 45:50:5 (v:v:v) water, acetonitrile, and formic acid (mobile phase B), with an elution gradient from 6 to 30% B for 10 min, 30 to 50% B for 9 min, and 50 to 6% B for 2 min, followed by 10 min of stabilization. The detection wavelengths were 320 nm for hydroxycinnamic acids (HCADs) and 360 nm for flavonols, using chlorogenic acid and quercetin as standards for external calibrations, respectively.The extraction procedure and HPLC analyses were performed according to Fritz et al., (2022)  using hiThe following reagents were added to a microtube in the following order: 15 µL of standard or extract, 750 µL of deionized water, 75 µL of Folin-Ciocalteu reagent, 300 µL of 20% m/v sodium carbonate, and 360 µL of deionized water. Solutions were incubated at 20 °C for 30 min in the dark; then, 250 μL of the solution was added to a 96-well plate, and the absorbance at 750 nm was obtained . Chlorog3 in a 10:1:1 v:v:v ratio. Then, 285 µL of FRAP solution was transferred to a 96-well plate, which was heated to 37 °C, followed by 15 µL of sample or TROLOX standard with an incubation time of 30 min. The absorbance was measured at 593 nm. The results were expressed as TROLOX equivalents. Lipid peroxidation was determined by the formation of malondialdehyde (MDA) using the methodology described by Du and Bramlage., (1992) [−1).To measure the antioxidant activity, the same extract used to measure the phenolic compounds in the leaves was used. The Trolox equivalents antioxidant capacity (TEAC), copper ion reducing antioxidant activity (CUPRAC), and the antioxidant activity by DPPH were evaluated . The ant, (1992) . The absSolanum tuberosum were statistically analyzed using a factorial analysis of variance (ANOVA). The data that did not meet the statistical assumptions of normality and homoscedasticity were transformed using the ln function, but the results were presented in their original measurement scale.The main effects of mycorrhizal inoculant and water stress applications and their interactions for each genotype of p < 0.05 was considered statistically significant. For the analyses, SPSS 22.0 software  was used.Treatments with significant differences were analyzed using Tukey’s HSD as a post hoc test to compare the means between treatments. The data were also subjected to a principal component analysis (PCA) to evaluate the multivariate effect of the established treatments and the relationship between the experimental variables. For all procedures, a Solanum tuberosum plant was evaluated. The objective was to analyze the physiological and metabolic responses of two genotypes with different colorations, with the VR808 genotype displaying white skin and yellow flesh and CB2011-104 displaying purple skin and flesh. The VR808 genotype had higher protection under the S1 stress level with both mycorrhizae, CC and HMC16, but separately. However, the CB2011-104 genotype under the S2 stress level had greater protection, and all the AMF were demonstrated to protect the plant from this level of stress. The different results indicate the different combinations of AMF, potato genotype, and stress level, which affected the content of phenolic compounds and antioxidant activity in the plant. Lastly, the effectiveness of using AMF to counteract drought stress, with interesting results in both genotypes, showed that CC and HMC26 mycorrhizae can defend potato plants that have been exposed to water stress. In potato crops, AMFs promote growth and increase tolerance to abiotic stress caused by water deficit, since they improve the morphology and antioxidant activity of these plants; however, these effects are linked to the selection and optimization of the inoculum of HMA; it is for this reason that inoculation turns out to be a promising future strategy for the cultivation of Solanum tuberosum.The response of two potato genotypes under deficit irrigation that was exposed during different stages of development of the


Plantar fasciitis (PF) is the most common cause of heel pain in adult. There are a variety of ways to treat PF, but these treatments have varied result in their effectiveness, and exist different degrees of limitations. At present, clinical studies focus on the effect of glucocorticoid (GC) and platelet rich plasma (PRP) in the treatment of PF, but there is a lack of systematic evaluation PRP and GC's clinical effect towards PF. This study aims to evaluate the long-term efficacy of GCs and PRP in the treatment of PF by means of meta-analysis.The literature of a randomized controlled clinical trial of PRP in the treatment of plantantifasciitis was searched on the Internet. Retrieve 7 databases. EndNote X9 software was used for document management. The Jadad scale was used to score the literature. Risk assessment of the literature was conducted according to Cochrane's systematic evaluation manual 5.0. RevMan5.3 software was used for literature risk bias analysis. Stata12.0 software is used for sensitivity analysis.This study will provide effective evidence-based evidence for the long-term efficacy of PRP and GC in treating PF.A systematic review and meta-analysis were conducted for the comparison of the long-term effect of PRP and GC on plantar fascia in the treatment of PF. The pathological mechanism of PF is unclear, but it has been widely accepted for medical workers that PF is caused by heavy load or excessive tension results in a slight tear in the fascia. And under continuous micro tears and chronic damage accumulation, the chronic inflammation of the fascia is building up. Also, some scholars believe that the emergence of the disease is closely related to obesity, standing up for a long time, incorrect walking posture, unfit shoes, the habit of work and study, and other factors.,6 According to epidemiological studies, the incidence of PF in the United States is about 10%, and it is estimated that PF accounts for 11% to 15% of all foot diseases. Besides, studies have found that most PF patients are aged between 25 and 64 years old, with the highest incidence between 45 and 64. Moreover, the incidence rate of athletes or regular runners and soldiers is significantly higher than that of the general population, which is about 4% to 22%.Plantar fasciitis (PF) is a self-limited disease characterized by pain in the medial plantar after getting up or prolonged sitting along with mild plantar flexion and varus in the foot during walking and localized tenderness in the medial side of calcaneal tuberosity.,11 However, although there are a variety of ways to treat PF, these treatments have varied result in their effectiveness, and exist different degrees of limitations. Therefore, it is very important to find effective treatment for PF.At present, the treatment of PF focuses on pain relief, and mainly adopt conservative treatment. Common treatment measures include having a rest, icing plantar locally, using non-steroidal anti-inflammatory drugs, adopting physical therapy, using foot orthoses, injecting corticosteroid, etc. The pathological manifestation turns to be rupture of fascia fiber and local aseptic inflammation. The common treatment in clinical is to inject glucocorticoid (GC) into the specific point on fascia. GC does show a distinguished inhibitory effects when it is up to physiological level. But in the late stage of GC treatment, it easily causes rupture of metatarsal fascia and atrophia of plantar fat pad. Focus on this question, the clinical discovered platelet rich plasma (PRP) promotes the proliferation of bone marrow mesenchymal stem cell, adipose mesenchymal stem cell, and tendon cell in plantar fascia, and accelerates tendon's repair. Which can be used in PF's treatment. Whereas, although there have been some randomized controlled trials (RCTs) of PRP and GC in the treatment of PF, there is a lack of systematic evaluation PRP and GC's clinical effect towards PF. To fulfill it, this study was conducted by meta-analysis, to compare the long-term effect between PRP and GC towards PF. Wish to provide evidence for clinical practice.The research shows that PF is a kind of local inflammation triggered by abnormal tension of both abnormalities of foot and fascia plantaris.22.1https://inplasy.com/inplasy-2021-10-0067/), and the systematic review protocol of us will be performed strictly following the guidelines of the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols (PRISMA-P) for systematic evaluation and meta-analysis.The protocol study has been registered on the Inplasy website Lack of PRP or adopted GC as intervention;(2)Adopted different basic therapy;(3)Final indicator does not include Visual Analogue Scale or Ankle Hindfoot Scale;(4)Repetitive contents;(5)Non-clinical RCT; non-human trial;(6)Baseline date (age etc) between 2 group shows a statistical difference;(7)Being evaluated to low quality research by Jadad scale.2.5First of all, eliminate repetitive contents been searched, conducted by EndNote X9. Then, exclude contents against standard after reading their title and abstract. Third, exclude contents against standard after reading the whole contents. Fourth, extraction of literature data. These data contain method of research design, interventions, methodology, baseline date (age etc) between 2 group, final indicator, follow-up and missing situation, etc. What have been mentioned above were conducted by 2 independent evaluators, and any differences that were difficult to determine could be solved by the third independent evaluator. The selection process will be shown in Figure Next, 2 independent evaluators will extract data from qualified literature according to a pre-designed data extraction table. The extracted content included author's name, year of publication, article title, sample size, gender and age of participants, diagnostic criteria, information about intervention and control groups, intervention measures, follow-up time, outcome indicators, and outcomes. Any disagreement will be resolved through consultation with a third independent evaluator.2.6 According to Cochrane Handbook 5.0, ① stochastic method; ② allocation concealment; ③ adopt blinding to volunteers and researchers; ④ adopt blinding to evaluator; ⑤ the completeness of research data; ⑥ selective reporting study outcomes; ⑦ other bias. To decide whether it is low bias risk, bias risk unsure or lack of information. What have been mentioned above were conducted by 2 independent evaluators, and any differences that were difficult to determine could be solved by the third independent evaluator.The “risk of bias assessment” tool recommended in Cochrane System Assessment Manual 5.0 was used to evaluate the included clinical randomized controlled studies.2.7Using Stata12.0 software and RevMan 5.3 software to do statistical treatment, outcome indexes performed in odds ratio. For the dichotomous outcomes, we will use the relative risk to measure the treatment effect, and for the continuous outcomes, we will use standard mean difference to analyze the effect. Both calculating 95% confidence intervals.2.82 test (test level α = 0.05) to assess statistical heterogeneity of the studies. When I2 < 50%, it indicates that there is a small statistical heterogeneity or no significant statistical heterogeneity between studies, using a fixed effect model; when I2 > 50%, it indicates that the data of studies exist considerable heterogeneity, applying random effects models to do combined analysis. When the heterogeneity is greater, the source of heterogeneity needs to be further analyzed.Adopting I2.9Galbraith radial plot and Egger test were used to evaluate the potential reporting bias of the inclusion study.2.10If there is a large heterogeneity between the studies, we will conduct a subgroup analysis to investigate the differences in age and sex, measure of intervention, etc. And we will also use Stata12.0 software for sensitivity analysis to assess the robustness of the study conclusions. If the results showed no qualitative change in the combined effect, the results are stable.2.11Using Jaded trial to evaluate clinical RCT's score. The total mark was 5, take scores below (include) 2 as low quality research, scores more than 2 as high quality research. Evaluation standard was: ① randomized blind: The total mark was 2, if there mentioned “randomized blind” and its synonymy gets 1 mark, any specific describe of achieving method about randomized blind add another 1 mark; ② withdraw and loss of follow up: total mark was 1, with explanation about withdraw situation gets 1 mark.2.12This study does not collect the personal information of clinical trial participants, so no ethical approval is required. The result of this research will provide reliable evidence-based medical evidence for the long-term efficacy of PRP and GC in treating PF, and the research will be published in peer-reviewed journals.3PF is the most common cause of chronic heel pain in adults, and its pathological manifestations are local aseptic inflammation and fascia rupture. There are many clinical treatment methods, among which GC and PRP are the common treatment methods for PF. However, there is still a lack of systematic evaluation of the clinical efficacy of PRP and GC in the treatment of PF. In this study, a systematic review and meta-analysis were conducted for the comparison of the long-term efficacy of the 2 drugs in the treatment of PF, hoping to provide evidence-based medical evidence for the treatment of PF and guide clinical decision-making.Conceptualization: Rongrong Ge, Jun Wu.Data curation: Bangmin Wang, Jiawang Jiang, Qianan Cao.Formal analysis: You Zeng, Yunfeng Liu.Methodology: Jiawang Jiang, Bangmin Wang.Software: Rongrong Ge, Qianan Cao.Supervision: Rongrong Ge, Bangmin Wang.Writing – original draft: Rongrong Ge, Bangmin Wang, Shuying Chen, Jiawang Jiang.Writing – review & editing: Rongrong Ge, Shuying Chen, Jiawang Jiang, You Zeng.


Phaseolus vulgaris L. is one of the most produced legumes worldwide and may be an alternative to reduce the environmental impact of meat production as a reliable source of high-quality protein. Plant growth-promoting rhizobacteria (PGPR) are emerging as a sustainable option to increase agricultural production. To understand the dynamics between plants and microorganisms, the culturable microbiota of bean roots was isolated and identified at distinct stages of plant development  and root compartments . Diversity and abundance of bacteria associated with root compartments differed throughout the plant life cycle. Bacterial plant growth promotion (PGP) and protection abilities  were assessed and associated with plant phenology, demonstrating that among the bacteria associated with plant roots, several strains had an active role in the response to plant biological needs at each stage. Several strains stood out for their ability to display one or more PGP traits, being excellent candidates for efficient stage-specific biostimulants for application in precision agriculture.Current agricultural methodologies are vulnerable to erratic climate and are dependent on cost-intensive fertilization to ensure high yields. Sustainable practices should be pursued to ensure food security.  The use of chemical fertilizers, pesticides, and irrigation has enabled a steep increase in food production, sustaining the world population in the last few decades, but with high impacts on both human and natural systems. The current world population is around 7.7 billion, and by 2100, the forecast is for a 54% increase, which is about 11.88 billion people [Phaseolus vulgaris L.) is one of the legumes most produced and consumed by humans, both dry and green, with an annual production in 2020 of 28 million tons (MT) and 23 MT, respectively [The common bean  is one of the possibilities to increase food security and food nutritional value, protecting the soil, maintaining the services it provides, and producing safer food for human consumption ,7.2) is considered to be one of the most crucial ecological services that microorganisms can provide to plants, thereby reducing the application of inorganic nitrogen fertilization [According to Zhang et al. , the miclization . Moreovelization ,13,14,15lization ,17,18,19lization . Hence, lization ,22. Indilization . For inslization .The significant role of microorganisms in agricultural productivity is an indication of their potential to support a more sustainable food production system. Hence, understanding the role of microbial communities deserves urgent attention, as it may provide information that can boost crop productivity.The impetus for this study came from the need to verify if: (1) the interaction between soil cultivable bacteria and plant root changes throughout the plant life cycle, and (2) the plant growth-promoting abilities of the root-associated cultivable microbiota differ among different developmental stages. These dynamics may be related to specific plant needs at each stage of plant development, allowing the use of cultivable bacteria in precision agriculture and inducing the optimized development at each stage of the plant life cycle. The hypotheses were evaluated by assessing alterations in the cultivable biodiversity of the microbiota associated with different root compartments of bean plants at different growth stages and by evaluating the ability of isolated microorganisms to produce PGP traits at each developmental stage.v/v) to prevent alterations in the community of microorganisms present in the soil . The addition of sand made the soil lighter, facilitating the collection of roots.Plants were grown in equivalent parts of soil and washed and autoclaved sand  were sown in early July 2020. Plants were grown in a greenhouse environment and watered with sterile deionized water twice a day, in the morning and evening, to maintain constant soil moisture. The average recorded temperature varied between 18 and 22 °C, with daily thermal amplitudes between 5 and 8 °C.The mixture was used to fill black 15 L plastic pots with 10 L of the soil mixture. In each pot, five bean seeds  were selected.Bacteria were isolated following the method described by Somasegaran & Hoben  with modA loopful of each isolate was used to inoculate a 5 mL YMB tube. Tubes were incubated at 26 °C for 2–5 days, depending on the growth rate of the bacteria, and 500 μL of the resulting cultures were transferred to microtubes. To each microtube, 500 μL of sterile glycerol (30%) prepared in YMB medium was added, and the microtubes were vortexed and stored at −80 °C.The 351 isolates were typed using BOX repetitive element-polymerase chain reaction (BOX-PCR) to screen for unique fingerprints before 16S rRNA gene amplification. Thus, duplicate genotypes can be identified, and only a representative isolate of each fingerprint pattern can be identified by 16S RNA gene sequencing. Isolates were inoculated on YMA, and single colonies were used to prepare a bacterial suspension in 50 μL autoclaved deionized water. Each PCR tube contained a mixture of 1 μL bacterial suspension and 1 μL BOXA1R primer (5′-CTACGGCAAGGCGACGCTGAC-3′) , as descThe PCR products were electrophoresed on an agarose gel (1.5%). Each gel (20 wells) contained two ladders , one negative control, and 17 samples. Electrophoresis was performed at 80 volts for 70 min. After staining the gel for 20 min in ethidium bromide, it was immersed in distilled water for 20 min to remove excess staining. The gels were then observed and scanned under ultraviolet light to obtain the individual typing profiles. GelCompar II  was used to calculate the Pearson correlation coefficient and analyze the clusters formed by applying the unweighted pair group method with arithmetic mean (UPGMA. A representative isolate from each fingerprint was randomly selected. The procedure yielded 227 representative isolates.The 16S rRNA gene from the representative isolates was amplified using primers 27f (5′-AGAGTTTGATCCTGGC TCAG-3′)  and 1492Agarose gels (1% agarose) were then electrophoresed. Each gel had 20 wells, two ladders, and one negative control, and the others were filled with samples. Electrophoresis was performed at 80 volts for 45 min, and the gel was stained for 20 min in ethidium bromide and then 20 min in distilled water to remove excess staining. The gels were then observed and scanned under ultraviolet light to obtain a clean bar at position 1400 bp in comparison with the NZYTech ladder control.The PCR products were sent to Eurofins Genomics  for sequencing. Each sequencing reaction tube contained the results of the PCR clean-up protocol or purification of DNA from enzymatic reactions using the NZYGelpure kit . Sequences (between 546 and 1140) were edited using FinchTV V1.4.0 . A BLAST search against the GenBank database was performed to identify bacteria at the genus level. Partial 16S rRNA gene sequences from the referred isolates were deposited in GenBank (ON419136-ON419292).Three plant growth-promoting (PGP) abilities were evaluated in the 227 isolates: siderophore production, indole-3-acetic acid (IAA) production, and antifungal capacity.To assess the ability of bacteria to produce siderophores, strains were grown for ten days on YMA medium supplemented with chrome azurol S (CAS) solution consisting of 1.21 mg/mL CAS, 0.1 mM FeCl3·6H2O, and 1.82 mg/mL hexadecyltrimethylammonium bromide (HDTMA). The presence of an orange halo around colonies was considered positive for siderophore production . Data arg and 4 °C, the supernatant was collected, and 500 μL of supernatant was reacted with 200 μL of Salkowski reagent  for 10 min at room temperature. The absorbance was measured at 530 nm using a spectrophotometer. IAA concentration was determined using IAA (Sigma) as a standard [To quantify IAA production, strains were grown in 5 mL tubes of yeast-mannitol broth (YMB) supplemented with 100 μg/mL of tryptophan, the precursor of IAA. Strains were grown in the dark at 26 °C and 150 rpm, until 1.0 optical density at 600 nm or when maximum growth was reached after five days. After centrifugation for 10 min at 10,000× Fusarium oxysporum L3W_8, GenBank Accession: OP071238) to guarantee that active hyphae were used. F. oxysporum was selected because it is recognized as one of the major causes of losses in agriculture worldwide in several different crops, including the common bean [Because bacteria exhibited slower growth than fungi, YMA plates were first inoculated with bacteria in the dark at 26 °C. The bacterial incubation time varied between 1 and 5 days depending on the bacterial isolate. When the colonies reached 0.5 cm, the center of the plate was inoculated with a 4 mm diameter plug from the peripheral zone of the fungal colony . Pearson correlation vectors of plant growth-promoting parameters (correlation ≥ 0.97) were specified as supplementary variables on the PCO graph, allowing the identification of the descriptors that contributed most to the differences observed among the stages evaluated. http://bioinformatics.psb.ugent.be/webtools/Venn/ (accessed on 1 January 2022)), to compare the presence of bacterial genera throughout the different stages of bean plant development.Venn diagrams were constructed using a tool from Bioinformatics and Evolutionary Genomics, University of Ghent . The four most common genera represented 61% of the identified diversity. The second group of the most represented genera accounted for 23.2% of the observed diversity. The least represented genera were Chitinophaga and Microbacterium with four strains; Rhizobium and Leclercia with three strains; Curtobacterium, Stenotrophomonas, and Mucilaginibacter with two strains; and Flexibacter, Azospirillum, Trinickia, Cronobacter, Pantoea, Klebsiella, Acidovorax, Cupriavidus, Agrobacterium, and Paraburkholderia with one strain each.The identified diversity of the cultivable microbiota associated with bean roots at different developmental stages is shown in Achromobacter, Flavobacterium, Bacillus, Priestia, and Pseudomonas. The genera Enterobacter and Delftia were recorded at all stages, except the pod stage (P). Variovorax was identified at all stages, except the late vegetative stage (V2). Chitinophaga were found at the flowering stage (F) and final vegetative stage (V2). Rhizobium was found at the pod (P) and late vegetative (V2) stages. Four genera were only present in the early vegetative stage (V1): Pantoea, Leclercia, Stenotrophomonas, and Klebsiella. Six genera  were exclusive to the late vegetative stage (V2). Five genera appeared exclusively in the pod stage (P): Paraburkholderia, Agrobacterium, Microbacterium, Acidovorax, and Cupriavidus. No genera were found to be associated solely with the flowering stage (F).Diversity throughout the plant life cycle  evidenceEnterobacter, Pseudomonas, Flavobacterium, and Bacillus, accounting for 72.1% of the total, with Enterobacter being the most represented genus in the three root compartments. Enterobacter (29%), Flavobacterium (21%), Pseudomonas (14%), and Bacillus (14%) were the most abundant genera in the nodules (N). Inside the roots (In), Enterobacter (33%), Pseudomonas (17%), and Delftia (16%) were the most abundant. Outside the roots (Out), Enterobacter (29%), Pseudomonas (20%), and Flavobacterium (20%) were the most represented genera.The diversity associated with different root compartments in plants at different growth stages is presented in Pseudomonas, Kosakonia, Enterobacter, and Flavobacterium were the most abundant genera, accounting for 55% of the total. Within this stage, the most represented genera were Pseudomonas (31%) and Flavobacterium (23%) for nodules, Kosakonia (27%), Enterobacter (23%), Pseudomonas (23%) inside the root, and Enterobacter (15%) outside the root. Higher diversity (12 genera) was observed outside the roots (9 genera) and nodules (6 genera).During late vegetative growth (V2), a higher diversity of bacterial genera (15 genera) was observed. Pseudomonas, Bacillus, and Flavobacterium, were the most abundant, totaling 67% of the strains. Within this stage, five genera were identified in the nodules, with Pseudomonas (38%) and Achromobacter (25%) being the most abundant; six genera inside the root, with Bacillus, Priestia, and Flavobacterium being the most abundant ; and seven genera outside the root, with Pseudomonas (38%), Bacillus (24%), and Flavobacterium (14%) being the most abundant.At the flowering stage (F), three genera, Pseudomonas and Bacillus were the most abundant (47% of the strains), followed by Priestia, Variovorax, and Microbacterium (33% of the strains). In the nodules, 50% of the strains belonged to the Pseudomonas genus. Inside the roots, three genera were more represented . Bacillus (22%) and Pseudomonas (17%) were the most represented genera outside the root, but their diversity was higher, and two more genera were identified.Thirteen genera were identified in the pod stage (P). Pseudomonas was common to all stages in the nodules. Flavobacterium was recorded at all stages, excluding the pod stage (P). Enterobacter was present only during the vegetative stage (V1 and V2). In contrast, the genus Variovorax was only recorded during the reproductive stages (F and P stages). In the nodules, 11 genera exclusive to one stage were found. Enterobacter was present at all stages inside the root (In), except for the P stage. Achromobacter and Pseudomonas were recorded in all stages, but not in the F stage. The genus Variovorax was identified in all stages, but not in the V2 stage. Bacillus was found in all stages, but not in V1. The genus Delftia was found at Stages V1 and V2. Chitinophaga and Flavobacterium were recorded in stages F and V2, respectively. The genus Priestia was observed in stages F and P. Two genera were exclusively found inside the root at V1, two other genera (Flexibacter and Kosakonia) at V2, and Microbacterium and Cupriavidus were specific to P. Outside the root (Out) Flavobacterium, Bacillus, Priestia, and Pseudomonas were present at all stages. Enterobacter was recorded in all stages, but not in the P stage. The genus Delftia was identified in Stages V2 and F. Fourteen genera were identified in only one stage, especially V2 and P.Analysis of each root compartment  evidenceThe IAA production ability of the strains differed with plant development a. SignifDifferences among developmental stages a showed Fusarium oxysporum growth. However, inhibition higher than 50% was registered at all stages, especially at the pod stage, with a significantly higher antifungal activity than in the first three growth stages. The overall analysis of antifungal capacity a showed A more detailed analysis highlighted the differences among the different root compartments for each stage b. At thePCO1 appeared as the main axis (69.7%), explaining the variation in the PGP ability of bacteria isolated from the roots of bean plants at different growth stages . Along PThe bacterial communities associated with plant roots can be influenced by several factors . In thisPseudomonas, Bacillus, Priestia, Variovorax, Enterobacter, Kosakonia, and Flavobacterium were the most common genera in the cultivable root microbiota of bean plants, and changes throughout the life cycle were noted. The genus Enterobacter was the most represented during the initial vegetative stage (V1). The genera Pseudomonas, Flavobacterium, and Bacillus were present throughout the plant life cycle, although differences in representativity among the stages were observed. Pseudomonas and Bacillus were more abundant in the reproductive stages (F and P). Flavobacterium was less abundant in the pod stage (P). In the vegetative stage, Kosakonia increased from V1 to V2. Indeed, the Kosakonia genus participates in some processes specific to this stage of development, such as root branching and growth, nutrient uptake, and water uptake, thus favoring vegetative plant development [The first hypothesis, that the diversity of cultivable bacteria associated with roots differs with plant development, which has already been hypothesized by other authors ,36, was elopment ,38,39. Priestia appeared to be highly represented in relation to the vegetative stages (V1 and V2). Variovorax was also highly represented in the pod stage. In contrast to the other three stages, the genus Enterobacter was not present at the pod stage (P). The apparent loss of Enterobacter during late bean plant development can be linked to alterations in roots at this stage [Enterobacter growth. In contrast, Bacillus increased steadily during the flowering and pod stages (F and P). According to Chaparro et al. [Pseudomonas, Flavobacterium, Bacillus, and Priestia. Although there were fewer isolates, several genera of bacteria were common in all four developmental stages. This basal microbiota may be complemented by other bacterial genera that are specific or more abundant at certain stages of plant development. The literature shows the capabilities of several strains of these poorly represented and sporadic genera to exhibit growth-promoting characteristics that may participate in plant development directly, through IAA or siderophore production, or indirectly, through protection against phytopathogenic fungi [In the reproductive stages (F and P), the genus is stage , possiblo et al. , it is pic fungi ,42,43,44Enterobacter, Pseudomonas, Delftia, and some other genera less represented. At V2, Delftia was replaced by Kosakonia. At F and P, Bacillus and Flavobacterium, which are minor genera, became dominant, and the genus Priestia also increased. Thus, in bacteria colonizing the root, major changes were observed between the plant vegetative (V1 and V2 stages) and reproductive (F and P stages) growth, possibly evidencing different growth requirements, more related to the development of root and photosynthetic organs initially and changing to flowering and seed growth later [Leclercia, Stenotrophomonas, Trinickia, Kosakonia, Curtobacterium, Azospirillum, Cronobacter, and Chitinophaga being exclusive to vegetative growth (V1 and V2) and Paraburkholderia, Rhizobium, Agrobacterium, Microbacterium, and Variovorax being exclusive to the pod stage.As an insight into the diversity in root compartments, the cultivable bacterial community present in nodules was much more specific and conserved throughout the plant life cycle, possibly indicating the particular conditions present in nodules where only some bacteria are able to grow. Inside the root, differences were observed throughout plant development, evidencing that conditions inside the root changed, initially supporting (V1) the growth of th later . OutsideAt the beginning of development (V1), the bacteria associated with the root may reflect the diversity present in the soil, and no major changes in diversity were observed within the three root compartments. Over time, the bacterial community may adapt to the different conditions present inside and outside the roots, favoring the genera most adapted to the prevalent conditions in each compartment, and the microbiota became more diverse among compartments . These vThe second hypothesis, that changes in diversity would reflect alterations in the PGP ability of strains meeting plant needs at crucial moments of its development, was also confirmed by our data .Pseudomonas, Enterobacter, Bacillus, Priestia, Achromobacter, Microbacterium, Curtobacterium, and Trinickia showed antifungal activity, confirming the information presented in the literature [Flavobacterium as a genus with high antifungal ability, with some strains inhibiting fungal growth above 60%.Antifungal activity was similar until the flowering stage  but increased significantly at the pod stage, validating the hypothesis that PGPR capacity varies throughout the plant life cycle. Differences among root compartments were also observed, with higher activity occurring inside the root (V2), outside (F), or not differing (V1 and P). In the early vegetative stage (V1), the root system is actively growing, and associations with microorganisms are being established . Thus, dterature ,51,52,53Priestia, Variovorax, Kosakonia, Leclercia, Pseudomonas, Enterobacter, Bacillus, Flavobacterium, Microbacterium, Rhizobium, Leclercia, Mucilaginibacter, and Cupriavidus. In addition to its use as a biostimulant, the capacity of the bacterial strains tested to synthesize IAA can be taken into account for the production of IAA, replacing the synthetic alternative and making the process more efficient and sustainable [IAA production was significantly different between the early vegetative (V1) and flowering (F) stages, but the results showed that the highest capacity was detected in isolates from the late vegetative (V2) and pod (P) stages, validating the hypothesis that PGPR capacity varies throughout the life cycle. Auxins are a class of phytohormones that regulate the entire period of plant growth and development in processes such as cell division, elongation, and differentiation . Accorditainable . Some getainable ,63,64,65Chitinophaga, Flexibacter, and Trinickia showed the ability to produce siderophores, adding new genera to the list that already included Pseudomonas, Enterobacter, Bacillus, Flavobacterium, Kosakonia, Variovorax, Delftia, Priestia, Achromobacter, Microbacterium, Rhizobium, Mucilaginibacter, Azospirillum, Acidovorax, Cupriavidus, Agrobacterium, Paraburkholderia, and Curtobacterium [Siderophore production may reflect the plant’s need for iron. In the late vegetative stage (V2), the bacteria in the nodules showed a high ability to synthesize siderophores. At this stage, the most effective nodules (higher and pinker) were observed. The effectiveness (and pink color) of nodules is linked with leghaemoglobin, a red-colored protein containing iron, responsible for controlling oxygen diffusion and protecting the oxygen-sensitive enzymes from denaturation, such as nitrogenase in bacteroids, but at the same time supplementing sufficient oxygen for bacterial respiration . Nitrogeacterium ,73,74,75The results from this study indicate a high cultivable diversity of genera associated with bean roots, which changes throughout plant development. Some genera were common throughout the plant life cycle, while others were specific to one or more developmental stages, and sporadic genera were also encountered.Higher bacterial specificity in the internal tissues was observed both in the nodules and in the internal parts of roots, indicating more constant and defined conditions inside the plant. The more diverse conditions outside the root certainly received the plant’s influence through root exudates but also from external pressures, supporting the higher bacterial diversity observed.Bacteria play important roles in the regulation and development of the plant life cycle. The variation in the observed diversity of bacteria associated with the bean root was reflected in the PGP ability of the strains, possibly meeting the plant needs at vital moments of its development, which was evidenced by the differences observed in bacterial PGP traits associated with each developmental stage.At each plant stage, several strains stood out for their ability to produce one or more PGP traits. Tailoring inoculants, not only to specific developmental stages but also to specific conditions and crops, can become a new option to produce biostimulants that are better adapted to plant-specific needs. Thus, propelling precision agriculture will contribute to more sustainable and environmentally friendly food production.


Metal foams possess remarkable properties, such as lightweight, high compressive strength, lower specific weight, high stiffness, and high energy absorption. These properties make them highly desirable for many engineering applications, including lightweight materials, energy-absorption devices for aerospace and automotive industries, etc. For such potential applications, it is essential to understand the mechanical behaviour of these foams. Producing metal foams is a highly challenging task due to the coexistence of solid, liquid, and gaseous phases at different temperatures. Although numerous techniques are available for producing metal foams, fabricating foamed metal still suffers from imperfections and inconsistencies. Thus, a good understanding of various processing techniques and properties of the resulting foams is essential to improve the foam quality. This review discussed the types of metal foams available in the market and their properties, providing an overview of the production techniques involved and the contribution of metal foams to various applications. This review also discussed the challenges in foam fabrications and proposed several solutions to address these problems.  Metal foams are cellular structures comprising solid materials with a large portion of gas-filled pores by volume. Due to their cellular structure, metal foams possess a set of unique mechanical and physical properties. These properties allow them to become highly efficient in several engineering applications, notably in components for blast resistance, fire resistance, thermal insulation, foam core sandwich panels, and sound and vibration damping ,2. In adThese foams are mostly developed by the addition of foaming agents or space holders in the matrix metal. Several methods have been used for foam development. These methods include adding foaming agent in liquid melt , compactFoam structures are divisible into two types, i.e., the open- cell foams and closed-cell foams, as listed in In open-cell foams, no film occurs between adjacent cells in the matrix material. A larger effective surface area (10–1000 times) is exposed to the surroundings compared to the dense material. They have sponge-like interconnected pores, as shown in Cells in the closed-cell foams are separated by a thin film of matrix material b 28,29],2928,29]2 as foaming agent. The resultant foams exhibited a uniform distribution of coated carbon fibre on the cell walls of aluminium foam, which means good wettability at the carbon fibre–Al foam interface and thus strong interfacial bonding of carbon fibres with the Al matrix was achieved. 2O3 particles were developed via melt foaming. These foams exhibited a closed cell, uniform cell size and roughly equiaxed polyhedral structure. In addition, the uniform distribution of the Al2O3 particles in the cell wall resulted in good interfacial interaction between the A359 matrix and Al2O3 particle [The microstructure of the cell-wall matrix influences mechanical properties and necessitates a controlled pore structure in metal foams. The microstructure analysis of Ni-coated carbon fibres reinforced AlSi7 foams was carried out. These foams were fabricated by a powder metallurgy technique using TiHparticle .The uniform cellular structure and dual-size cellular structure  under compression as shown in Using potassium carbonate particles as space holders, copper foams were developed by the powder metallurgy technique. The two types of pore structure were formed: macro-pores and micro-pores. Macro-pores were interconnected spherical-shaped pores with the same size as potassium carbonate particles, while micro-pores were found in the pore struts and walls. The open-pore structure and connectivity of the macro-pores are formed by these micro-pores, resulting in foams of a high degree of pore connectivity . The homThe selection of a proper compaction pressure is critical for producing high-strength foams because it affects their pore morphology. The influence of compaction pressure on pore morphology and densification was analysed using acrawax as space holders, which are lubricating and compressible. At lower compaction pressure (200 MPa), spherical pores were obtained, however, the cell walls were porous (micro-pores). Denser cell walls and mostly spherical pores were obtained when compacted at 300 MPa. At higher compaction pressure (400 MPa), the cell walls were more dense, but the high pressure deformed the pore morphology into an ellipsoidal shape . The stu4 phase. The cooling curve of Al foams showed that during solidification, primary α-Al formed first, followed by CaAl4; thus, CaAl4 was not present in the melt and does not contribute to thickening [The effect of anodization treatment on the morphology of primary α-Al grains, as well as the constitution and distribution of secondary phases induced by adding thickening and foaming agents to Al foams developed by melt foaming, were investigated. The results showed the formation of dendritic α-Al grains and CaAlickening . In addiickening ,56,57,582 particles to Al foam increases the maximum foam expansion significantly due to the solid phase lowering the minimum cell wall thickness, allowing larger expansions before cell rupture and collapse. This results into higher proof stress, yield strength and more energy absorption for a given strain as compared to pure Al foams [The mechanical properties of metal foams, particularly their compressive strength and energy absorption capacities, are dependent on their cell structure, porosities, and relative densities. Al foams ,65. −3. The energy absorption capacity has the highest values of 72.22 MJ m−3 at 45.8% porosity. In addition, the compression properties of AlCu5Mn foams were found to be better than those of other Al-based foams [The higher values of porosities can reduce energy absorption capacity of foams. Therefore, it is essential to develop foams with optimum porosities. As the porosity of AlCu5Mn foams increases from 45.8 to 91.2%, the corresponding energy absorption capacity decreased from 72.22 to 2.70 MJ med foams . The effed foams . In anoted foams . The comed foams . The comed foams . The angIn an Al foam prepared by the space-holder technique, the plateau region as shown in f of metal foam.pl = plateau stress of the metal foam, σys = yield stress of the cell metal foam; ρrel is the relative density; C = shape factor = 0.3 Ef = elastic modulus of the metal foam, and Es = elastic modulus of the solid metal. As shown in −1, the shape factor values were found to be C = 0.33, 0.42, and 0.48, respectively. In addition, for Ef shape factor, values were 0.52, 0.62, and 0.72 at the strain rates of 0.001, 0.01, 0.1 s−1, respectively. The value of ‘C’ was found to increase with strain rate. Furthermore, Jiang et al. [The mechanical properties of metal foam have been predicted using Gibson–Ashby mathematical models ,85,86. Ag et al.  developeSimilarly, Bekoz and Oktay  developeAs it is clear that the plateau stress and yield strength are a function of relative density , Figure Metal foams are primarily manufactured using either liquid-phase (or melt foaming) or solid-phase  techniques.2O3, etc.) [Melt foaming is particularly popular because it produces relatively inexpensive foams with desirable properties ,97. The 3, etc.) ,99. In m3, etc.) .2 [3 [3)2) [2, CaO, Al2O3, etc., are also added into the melt [2 and releasing gas and bubbles. The foamed melt is taken out of the furnace and cooled in the air. This technique was used to fabricate A356/20SiC composite foams with varying porosities and cell sizes with TiH2 as a blowing agent [Some of the common foaming agents and injection gases include TiH2 ,104,105,2 [3 ,107, zir2 [3 ,109, dol[3 [3)2) ,111, etcthe melt ,112. Theng agent .3 as a blowing agent and SiC as a foam stabiliser. The yield stress and collapse plateau stress of composite foams increased along with the SiC volume fraction [2 were developed to investigate the effect of the thermal decomposition of TiH2 on the foaming behaviour of the Al alloy. The effectiveness of the melt foaming of the Al alloy is highly dependent on the decomposition properties of TiH2 [3, TiH2, or MgH2 powders as blowing agents, which release gas during decomposition and gradually foam the magnesium alloy melt [2 and CaCO3 were mostly used as foaming agents for the development of a foam structure.In another study, AlSi9Mg/SiC composite foams were successfully developed using direct melt foaming with CaCOfraction . Similarfraction . In addifraction . Additio of TiH2 . Mg alloloy melt ,119,120.loy melt . Several2 powder) or space holders in a powder mixer. The mixed powders are then compacted cold, using a uniaxial compaction, in a lubricated tool-steel die at required pressures to achieve precursor green densities. The precursor specimens are later led to a furnace, so that the foaming procedure take place under high temperatures in case of green compacts with foaming agents, while for compacts with space holders, porosities are acquired by a leaching process followed by sintering.In this technique, the metallic powder was mixed with some mass fraction of foaming agent (TiH2 or MgH2) were used to fabricate ZnAl4Cu1-alloy and AlSi12 alloy foams [Metallic powders and a foaming agent such as titanium hydride powder are mixed and then cold compacted, as illustrated in oy foams . It was oy foams .2CO3 [The space-holder technique enables producing metal foams with controlled pore morphology . Figure 2CO3 , carbami2CO3 ,139, car2CO3 , polymet2CO3 . Approxi2CO3 ,143. The2CO3 . Many authors used this technique to produce metal foams focusing on processing parameters, feasibility, and efficiency ,146,147.In addition, the advantages and disadvantages of these techniques are mentioned in Metal foams encompass a wide range of applications as mentioned in Another closed-cell Alporas (Al foam core) was fabricated into a composite beam a by embe2 of Ni foam annually for these applications [4HCO3) as a space holder was developed by powder metallurgy for bone implant application [Alcoa (the USA) developed a new method based on the continuous casting technique to reduce the cost of Al foams for inexpensive products . The tecications . Ti-baselication .The technology of metal foaming is developing at an accelerated pace with considerable progress. This research area involves interdisciplinary collaborations among physics, chemistry, and materials engineering to produce the required quality economically with reproducible foamed materials that possessed a unique range of properties. These diverse applications allow the construction of functionally new parts or devices. Since foams have many competitors that are mostly less expensive, it becomes crucial to improve fabrication technologies for large-scale foam production, generating a greater variety of low-cost metal foam products. Despite the successful development, more basic research remains essential to understanding the correlation between the composition of metal alloy and foamability. Producing foams of consistent quality, morphology, and control of structure is challenging. However, the foam quality and properties can be controlled by optimizing the factors and parameters. In addition, the foam’s porosities can be controlled by heat treatment or coating the blowing agent to stop the decomposition of the gas-blowing agent in the melt. Overall, to produce uniformly structured metal foams, it is necessary to optimize the processing parameters in the existing fabrication techniques. In addition, further studies are required to compare the available processes to produce metal foams from raw material (powders) or Al scrap. The development of alloy foams for the core with improved properties may contribute to their successful usage in various applications.The academic and industrial research plays a crucial role in eradicating problems that otherwise limit the wider applications of metal foams. Improvement is required to ensure the reproducibility and uniformity of the foam’s cell structures for attaining even densities and distribution of pore sizes in the entire part or component, resulting in uniform foam structures. Additionally, new processing routes, cheaper raw materials, and less wastage may reduce the processing and material costs of metal foams. Case studies based on simulation, innovative design, and testing could show end-users that despite the higher costs of some foams or parts, there are more benefits associated with these new structures and materials, such as savings in weight and energy.This review presents an overview of the metal foams developed by various techniques to enhance their properties and performance, summarising the efforts to improve the foaming behaviour and characteristics. Depending on their applications, foam structures could be tailored to generate open- or closed-cell foams. Microstructural studies evaluated the effect of reinforcements and fabrication techniques on the foams’ microstructure, and it defines their mechanical behaviour. The foams’ properties, particularly the compressive strengths and energy absorption, could be improved by adding different reinforcements, such as metal or ceramic particles, to stabilise the foam structure. The porosities in these foams are dependent on foaming agents and space-holder particles. The processing techniques that fabricate metal foams play a vital role in deciding their properties and foaming behaviour. Their applications as structural components in automotive parts, such as ships and aerospace transportation, and as sound dampers, filters, electrodes, etc. can be effectively attained. However, improvement would be possible by identifying the downsides and feasible solutions for better performances under various working conditions and production techniques. Although metal foams find applications in many sectors due to their lightweight and high strength, the high cost of fabrication techniques hinders large-scale production. The challenge of future process development should therefore focus on reducing the production cost as well.


The COVID-19 pandemic have had deleterious effects on mental health of students. Authors suggest that the psychological effects will persist long after COVID-19 has peaked, but we have no data to confirm this.Objective: The objective of this study is to compare clinical issues  and adjustment (coping strategies) in French university students during different phases of the COVID-19 pandemic in 2020 and 2021 (during two periods of lockdown and two periods after lockdown)T1 = 1294); during the period after lockdown ; 1 year after the first lockdown, which was also a lockdown period ; and 1 year after the first unlockdown, which was also a unlockdown period . The following variables were measured: concerns, coping strategies, anxiety and depressive symptoms.Method: Data were collected anonymously at four timepoints: during France’s first national lockdown (23 April- 8 May 2020; nIn 2021, students have significantly higher levels of anxiety and depressive symptoms than in 2020, and this is even more pronounced during the lockdown periods. For example, 44.1% had probable anxiety symptoms in the 2021 lockdown, compared to 33% in the 2020 lockdown. In the unlockdown periods, the rates are 21.7% in 2020 and 26.4% in 2021.Our results suggest that university students, known to be a vulnerable population with significant mental health deterioration, have become even more vulnerable with the COVID-19 pandemic.No significant relationships.


DNA methylation is an epigenetic mark that living beings have used in different environments. The MTases family catalyzes DNA methylation. This process is conserved from archaea to eukaryotes, from fertilization to every stage of development, and from the early stages of cancer to metastasis. The family of DNMTs has been classified into DNMT1, DNMT2, and DNMT3. Each DNMT has been duplicated or deleted, having consequences on DNMT structure and cellular function, resulting in a conserved evolutionary reaction of DNA methylation. DNMTs are conserved in the five kingdoms of life: bacteria, protists, fungi, plants, and animals. The importance of DNMTs in whether methylate or not has a historical adaptation that in mammals has been discovered in complex regulatory mechanisms to develop another padlock to genomic insurance stability. The regulatory mechanisms that control DNMTs expression are involved in a diversity of cell phenotypes and are associated with pathologies transcription deregulation. This work focused on DNA methyltransferases, their biology, functions, and new inhibitory mechanisms reported. We also discuss different approaches to inhibit DNMTs, the use of non-coding RNAs and nucleoside chemical compounds in recent studies, and their importance in biological, clinical, and industry research. DNA methylation is an essential epigenetic mark that living beings have used to survive in different ambient conditions. For example, in prokaryotes, it is used to differentiate their own DNA from foreign DNA and avoid endoreduplication . MoreoveThe MTases family catalyzes DNA methylation, and it is a conserved mechanism from archaea to eukaryotes, from fertilization to every stage of development and from early stages of cancer to metastasis . HoweverThis work focused on DNA Methyltransferases, their biology, their functions, and their new inhibitory mechanisms reported. Many studies have focused on DNMTs because they have an important role in the epigenetic regulation of living beings . Other iDNA methyltransferases (MTases) are conserved in living beings acting in orchestra with other epigenetic players. They have positioned themselves with a few exceptions as the main transcription regulators. MTases are a group with methyltransferase activity; they have evolved in different orthologs, but all have the methyltransferase domain and a DNA target recognition domain. Living beings have conserved MTases to survive the different and dynamic ambient conditions ,3,4,5,19l-homocysteine (AdoHcy) and methylated DNA [The reaction mechanism of MTases catalyzes DNA methylation in adenine or cytosine bases is known. All MTases interact with the cofactor S-adenosyl methionine (AdoMet) to transfer a methyl group and produce S-adenosyl-ated DNA . Moreoveated DNA   was the first DNMT identified with DNA methyltransferase activity. DNMT1 is called maintenance DNMT because it methylates hemimethylated DNA through the cell cycle to maintain the epigenetic memory in differentiated cells ,26,27. DDrosophila melanogaster [Saccharomyces cerevisiae, Saccharomyces boulardii, Schizosaccharomyces pombe [Caenorhabditis elegans  system, which consists in that MTases methylate host DNA and do not methylate foreign DNA . It is tvolcanii , Pyrocucs abyssi , Pyrococrikoshii ,67, Sulfctaricus , and Thetolerans . In bactbacteria , Borreligdorferi , Escherihia coli , Helicobr pylori , and als species .In protists, we know that there are 5′ cytosine methyltransferases homologs because they have been reported by analysis in silico ; howeverVolvox carteri, Methyltransferase1 (met1), is a homolog of DNMT1 in mammals and is detected during the DNA replication fork. It has been localized in transposon CpG sequence methylation [met1) Chromomethylases (CMT), Domains Rearranged Methyltransferase (DRM). In algae, MTases are overexpressed during the sexual and asexual phases of the green algae Boechera genus [Algae and plants have DNA methylation and the same MTases homologs. In algae, MTases have been reported in the multicellular green algae. The MTase gene in hylation . Nowadayra genus . These fArabidopsis thaliana and Salvia milthiorrhiza act in gene methylation of non-coding RNAs and coding RNAs; also, in plants, MTases act in the phenomena of genomic imprinting, calling of epi-alleles [Cynara cardunculus var. escolymus development [Solanum pennellii [Elaeis guineensis, and plants detoxification of pesticides [MTases in plants such as -alleles ,43,74. R-alleles ; they arelopment ; in the ennellii , stress ennellii ; in prodsticides . These sHumicola grisea [Aspergillus clavatus [Aspergillus flavus [Fungi is a biological kingdom that has let us see the importance of metabolite production and the diversity of various types of metabolisms that have evolved to survive on every external condition. Although in fungi , homologa grisea ; the proclavatus , and thes flavus . Fungi aIn animals, MTases are DNA methyltransferases (DNMTs), which have been reported in several functions on invertebrates and vertebrates. DNMTs are present in all animal phyla. DNMTs are functional in radial animals, echinoderms: starfish and sea urchins. Interestingly, not all DNMTs are present in echinoderms; they have only DNMT1 and DNMT3. Another intriguing enigma to research is why DNMT1 does not have a PCNA binding domain in starfish . AnotherCrassostrea gigas treated with 5-azacitidine, an inhibitor of DNMTs catalytic activity [Aplysia is treated with RG108, another compound that inhibits DNMTs. This treatment was applied for 24 h and resulted in amnesia until 48 h [Solenopsis invicta, DNA methylation and DNMTs expression of different embryos and adults changed eight-fold [DNMTs are essential in the epigenetic memory of animals. Different types of memory in cells have been affected when DNMTs are inhibited. For example, early development is affected in the Oyster activity ; on the til 48 h . Moreovetil 48 h , fecundight-fold ,87 and sght-fold ; the excght-fold ,89.DNMTs are involved in vertebrate evolution and have been studied since embryonic development. In addition, it has been shown that DNMT3A leads the embryonic development . For exaOreochromis niloticus [Solea seleganensis, which has five paralogs of DNMTs that are regulated differentially during development. In addition, it is a model where it has been demonstrated that DNMT expressions change with temperature and 5-aza-2′deoxycitidine treatment [In the zebrafish genome, 8 DNMTs have been recognized: one DNMT1, one DNMT2, and six DNMT3 , and theiloticus . Other ereatment . Moreovereatment .In summary, several studies have shown the structural and functional role of DNMTs in vertebrates during development . DNMTs iTo know the regulatory mechanisms, we have focused on splicing alternative isoforms. The alternative splicing process post-transcriptionally regulates DNMTs. DNMT1 and DNMT3A produce catalytically active isoforms, and DNMT3B produces catalytically active and inactive isoforms ,99. OstlDNMT3B isoforms could be classified into three families. The first includes the isoforms DNMT3B1-7, which are the better studied. DNMT3B1 and DNMT3B2 are the catalytically active isoforms of DNMT3B; although DNMT3B2 lacks exon 10  Figure , it is kDNMT3B3 is a DNMT3B isoform that lacks exons 10, 21, and 22. But, it still possesses the methyltransferase motifs I, IV, VI, IX, and X, the target recognition domain, and 9 amino acids of motif XI in the carboxylic domain . DNMT3B3ΔDNMT3B is the second family of DNMT3B isoforms. This family has seven members reported. They do not have the first exons before the PWWP domain . For exaHelicobacter pylori infection. DNMT3B4 is also overexpressed in hepatitis, cirrhosis, and liver cancer cells [The alternative splicing process regulates DNMT3B to produce several isoform products. These products have a role in mediating DNMT methylation patterns. Each DNMT3B isoform has a cell-specific role in maintaining the optimal function of DNMTs in the tissue, and transcriptional deregulation could be a part of cancer development. Overexpression of each DNMT3B isoform has been associated with any cancer initiation, development, progression, or metastasis. Probably, Isoforms are defined by tissue origin, embryonic origin, and external factors by which cells are exposed . Therefoer cells . On the er cells , and oveer cells ,114.The variation of DNA methylation is essential in embryonic development. In fact, mutation of the catalytic domain of DNMT3B produces the syndrome of immunodeficiency facial instability 1 (ICF1) . The expOryzias latipes) have shown that DNA demethylation in sperm cells is during maturation. However, DNA methylation of the cells shows a gradual de novo methylation, after fertilization, until the finish of the gastrula stage. In the same work, DNMT3B was measured, and they found that the decrease in DNMT1 and increase in DNMT3BB.1 are associated with DNA methylation patterns. In Zebrafish, the authors show that the phenomena are similar but have dramatic de novo methylation from late morula to gastrula. In mammalians, at this point, there are variations too. In a mouse model (Mus musculus), during maturation, sperms gain DNA methylation. After fertilization, the demethylation process starts until the blastocyst stage, and it occurs dramatically de novo methylation, compared with humans (homo sapiens) [Importantly, it has been proved that germline mutation in the catalytic domain of DNMT3B has, as a consequence, the ICF1 syndrome in humans ,121. Thesapiens) ,123. In contrast, human sperm DNA is demethylated during maturation and fertilization; the zygote demethylates until blastocyst and then starts the de novo methylation process . These wDNMT3A gene [DNMT1 are related to genome-wide alterations of DNA methylation in colon cancer [The dysregulation of DNA methylation has a role in the development of cancer cells and other diseases ,124. As T3A gene , whereasn cancer   and 5-aza-2′-deoxycytidine (DAC or decitabine). These compounds are used for treating myeloid dysplastic syndrome (MDS) and acute myeloid leukemia (AML) ,130,131.More recently, a series of new quinoline-based inhibitors have been discovered. These belong to the non-nucleoside inhibitors, the first of them is SGI-1027 , from whOn the other hand, it has been reported that ncRNAs, which comprise small interfering RNAs (siRNAs), microRNAs (miRNAs), and long non-coding RNAs (lncRNAs), are capable of inhibiting DNMTs . FirstlyCCDC26 (a lncRNA that interacts with DNMT1) is lost, DNMT1 is mislocalized to the cytoplasm, turn causes the hypomethylation of the DNA [DACOR1 in colon cancer [HOXD-AS1 in lung adenocarcinoma [On the same line, miR29b is a miRNA that targets DNMT3A, and DNMT3B is downregulated in cancer of lymphoma and cancer of the pancreas ,143. In  the DNA . Other ln cancer  and HOXDarcinoma . linc-POarcinoma . AlthougCancer cell development has been associated with epigenetic alterations, and DNMTs are one of the main actors in transcription regulatory processes. DNMTs are regulated by several mechanisms as ncRNAs and deregulation of DNMTs have been found in cancer. These deregulations have been reported in several types of cancer in a combinatorial manner. That is why the regulatory mechanisms of DNMTs are important in the cancer study. However, studying not only the mechanism of action of the DNMTs but also their broad diversity in organisms and how they have evolved in time allows us to understand better how they are regulated and predict the consequences of mutations and deregulations. Furthermore, manipulating DNMTs expression could be of great importance to knowing the drugs and the different ncRNAs that can help regulate them in cancer cells.Our understanding of the family function of DNMTs has increased considerably as their discovery about thirty years ago. It is now clear that these proteins play critical roles in setting DNA methylation patterns genome-wide at specific developmental time points, particularly during early development and in specific tissues undergoing dynamic methylation in living beings. Most recent studies indicate that DNA methylation plays a role in variation during evolutionary context and adaptation to the ambient conditions. Other studies have shown their importance in clinical and industry research. Future biochemical and structural studies should focus on DNMT and DNMT isoform-specific and their relation with ambient variables to assess phenotypic association and elucidate functions in different types of cells, parasites, fungi, plants, and animal models. More studies are also necessary to understand the role of DNMTs isoforms in cancer and its interrelation between tumors and the origin tissues.


Chronic pain, nocturnal cramps, and sleep alterations are prevalent symptomsand signals in Charcot-Marie-Tooth disease patients. Sleep and pain arebidirectionally related and physical therapy can improve the binomial sleepand pain/nocturnal cramps. Therefore, we hypothesized that the applicationof a specific physical therapy program for Charcot-Marie-Tooth disease wouldimprove sleep quality, pain perception, and nocturnal cramps.A non-randomized controlled study that included 9 Charcot-Marie-Tooth diseasepatients  and 8 controls(active control group - booklet on sleep hygiene). The intervention lasted 8weeks, three sessions per week. The effects were evaluated ten days before(baseline) and ten days after the intervention (post). Our primary outcomewas sleep quality ; and secondary outcomes werepain perception (brief pain inventory) and nocturnal cramps(self-report).p=0.005) and nocturnal cramps(p<0.001) but had no effect on actigraphy data(p>0.05) neither on pain perception(p>0.05).The program was able to improve the subjective sleep quality(Our initial hypothesis was partially corroborated: the improvement insubjective quality of sleep and nocturnal cramps is already beneficial forthe health promotion of the volunteers in this study affected by thedisease. Our findings may serve as a basis for future research to develop aprogram focused on the treatment of analgesia, which can improve painperception and alter the objective quality of sleep. It is aprogressive disease that initially affects the lower limbs with symptoms thatinclude feet deformity and leg atrophy1,5.Charcot-Marie-Tooth disease (CMT) is a hereditary peripheral neuropathy,characterized by sensory and motor manifestations6.It affects mainly the extremities of the body (feet and hands) and can also manifestitself as muscle cramps6,7, which in CMT disease occurs mainlyat night8. These nocturnal symptomscompromise their quality of life and interfere in daily activities such as sleep andability to perform exercises8.Another symptom of the CMT disease is the neuropathic pain that affects 23 to 100% ofthe individuals with the disease9demonstrated that about 63% of the pain cases in CTM disease are neuromuscularrelated, with an average duration higher than 15 years, which indicates a chronicpain condition.The study by Ribiere et al. (2012)10,11. In a previous study conducted by our group, weobserved that pain perception, fatigue, and nocturnal cramps were related to sleepalterations in patients with CMT disease12 and that these patients present greater sleep fragmentationand changes in its architecture13.Chronic pain and sleep are bidirectionally related: proper sleep improves physicaland psychological symptoms of pain and pain worsens sleep quality, but themechanisms involved in this relationship are not well established14 and is also used in sleep medicine as acomplementary treatment for several respiratory and neurological-baseddisorders15.Given this evidence, we hypothesized that the application of an intervention capableof improving pain symptoms could improve sleep and, consequently, improve thepatients’ quality of life. Physical therapy is indicated and used to treatneuropathies as it can promote functionality gainTherefore, physical therapy could contribute positively to the sleep-pain binomial inpatients with CMT disease, allowing them to recover their physical functionality andimproving their ability to perform daily tasks.The objective of this study was to evaluate the effects of a specific physicaltherapy program directed to CMT disease (SPTP-CMT) on sleep, pain perception, andnocturnal cramps in patients with Charcot-Marie-Tooth disease. We hypothesize thatphysical therapy will be able to improve sleep quality (subjective and objective),pain perception, and nocturnal cramps in these patients.A non-randomized controlled trial conducted between October and December 2017 inthree cities of Sergipe - Brazil: Tobias Barreto, Pedrinhas, andCristinápolis. Volunteers were recruited in partnership with the publichealth system and the study only began after approval by the ethics committeeaccording to Resolution No. 466/12 (No. CAAE: 69490117.5.0000.5546). This study wasregistered in the Brazilian Registry of Clinical Trials (RBR-6hyt5g7) and its designand conduct following the methods described by CONSORT.Inclusion criteria: individuals aged between 16 and 65 years, with aclinical and electrophysiological phenotype consistent with the disease. Thediagnosis was made by clinical and electroneuromyographic examination, in additionto the evaluation of the CMTNS score16. The control group was composed of individuals from the samefamily not diagnosed with the disease, matched for sex and age.Exclusion criteria: acute or chronic lung disease, cognitive orpsychiatric disorders, severe or poorly controlled hypertension, heart failure,chronic kidney disease, systemic diseases, diabetes mellitus, pregnant women,obesity, active smokers, alcoholics, and refusal to participate in the study.Sample size calculation: we performed a sample size calculation usingthe GPower 3.1.9.7 software, considering an effect size of 33%,power of 80%, and alpha of 5%, with two timepoints and two evaluation groups,obtaining a minimum value of 22 individuals (11 per group).16: resultsbelow 10 were considered as mild, between 11 and 20 points as moderate and above21 as severe.An initial assessment was performed to collect demographic data such as name,gender, date of birth, age, address, investigation regarding the use ofmedications, associated diseases, and anthropometry. A targeted CMT diseasescore was recorded to assess disease severity as described by Shy et al.(2005)Primary outcomes: subjective and objective sleep quality.Secondary outcomes: pain perception and nocturnal cramps.Nocturnal cramps: was based on self-report, the volunteers received anevaluation form in which they had to rate the intensity of the cramps from 0to 10.15. Resultsless than or equal to 5 were considered good sleep quality and greater than5 poor sleep quality. Was applied the version validated forPortuguese-BR17,18.Pittsburgh sleep quality index (PSQI): subjective assessment of sleepquality17,19. Scores less than 10 were considered as normal daytimesleepiness, and greater than or equal to 10 as excessive daytime sleepiness.Was applied the version validated for Portuguese-BR19,20.Epworth sleepiness scale (ESS): evaluates the chance of sleeping in everydaysituations19. Results equal to 0 wereconsidered as no pain; between 1 and 4 points as mild pain; between 5 and 6points as moderate pain and between 7 and 10 points as severe pain. Wasapplied the version validated for Portuguese-BR21,22.Brief pain inventory (BPI): assesses the perception of pain23,24.Chalder fatigue scale (CFS): assesses the perception of fatigue. Resultslower than 4 were considered as absent fatigue and higher or equal to 4, aspresent fatigue. Was applied the version validated forPortuguese-BR25 and Sadehet al. (1994)26, availablein the programs Action 3 - Version 3.15 and Action for Windows - Version1.05 . The estimated parameters were sleep latency, sleepefficiency, total sleep time, and wake time after sleep onset (WASO).The actigraph model used in this study was the Mini Motionlogger Actigraph -Basic 32  with the ZCM algorithm (“zerocrossing mode”). The time of collection was set at 1 minute. Thecomputerized analysis was done according to the algorithms proposed by Coleet al. (1992)Actigraphy data analysis was carried out by an individual with expertise inthe field for at least ten years and reviewed by the second author of thispaper, who is also an expert in the analysis of this type of data. Duringthe recording period, the volunteers also filled out a sleep diary used forcombined actigraphy analysis.Individuals participated in a specific physical therapy program for CMT disease(SPTP-CMT). The program was divided into two phases and consisted of exercisestargeting four domains: A) gain of joint range of motion in lower limbs; B)muscle strengthening in proximal and mid-limb segments; C) coordination, staticand dynamic balance; and D) functional independence. The intervention lasted 8weeks, totalizing 24 sessions, which were held three times a week, each sessionlasting an hour.Before the physical therapy sessions began, the volunteers performed an initialwarm-up series: going up and down a ramp five times and pedaling on an exercisebike for five minutes. Then, in a sitting position, they received pressure onthe soles of the feet with a six-centimeter diameter proprioception ball torelease the plantar fascia.The 24 sessions were divided into two exercise phases: phase 1 up to the tenthsession) and phase 2 (remaining fourteen sessions).A) Stretching (performed in three sets of 15 seconds for eachmuscle group).Hamstring stretching performed with the volunteer in dorsal decubitusposition with a stretching band supported on the midfoot for hip flexionwith knee extension. The contra lateral limb remained supported in flexionon the stretcher.Stretching of eversors, inversors, plantar flexors, and dorsiflexors: sittingon a chair, the volunteers performed the self-stretching with the stretchingband of eversors, inversors, plantar flexors, and dorsiflexors.Triceps sural stretching: the volunteer stood in a standing position with thehip and knee in extension and the foot resting on the floor, the oppositeside with knee flexion and the hands resting on the wall.Iliopsoas stretching: the last stretching series performed was the standingquadriceps and iliopsoas stretching, with knee flexion, gluteal contraction,and support for execution.B) Muscle strengthening (performed in three sets of 10repetitions).Gluteal strengthening: the volunteer was in dorsal decubitus, performing hipelevation with bent knees and feet supported on a stable surface(bridge).Strengthening of dorsiflexors and eversors: the volunteers were in a sittingposition and performed the exercise from the movement against elastic bandresistance.Strengthening the quadriceps: in the sitting position, the volunteersperformed a knee extension with a shin pad.Ischiostibial strengthening with the volunteer in standing position, kneeflexion was performed, with shin pads.C) Coordination, static and dynamic balanceProprioception exercises on a proprioceptive board, standing up, performingplantar flexion, and dorsiflexion for two minutes. Next, the unipodalsupport on a stable surface with and without visual feedback and, finally,the exercise of sitting and standing on a Swiss ball/chair, with or withoutsupport.A) Stretching (performed in 15 seconds standing + 15 secondswith torso flexion).Triceps sural and hamstring stretching: the volunteer was positioned standingon an incline.Quadriceps and iliopsoas stretching: the volunteer stood with one of thelower limbs on the stretcher, with the knee flexed, and the contralaterallimb with the foot on the floor and the knee flexed.B) Muscle strengthening (performed in three sets of 10repetitions).Strengthening the quadriceps, gluteal muscles, and posterior thigh: with theindividual lying in a supine position in dorsal decubitus with knees bentand support of the plantar part of the feet on the stretcher raising theglutes on the stretcher.C) Coordination, static and dynamic balancePlantar flexion and dorsiflexion for two minutes on a proprioceptivestanding board.Squatting with support of only one upper limb (if you could performwithout support) on foam with a density of 60 cm,Stepwise advance on a proprioceptive disk.Baseline (ten days before the beginning of the intervention)On day 0 (D0) the volunteers filled out the questionnaires, started theactigraphy, received the sleep diary. After ten days, the actigraphs wereremoved, sleep diaries collected, and the intervention started (D10). Thecontrol group also received a lecture on sleep hygiene and printed material onthe subject. The research team contacted the members of this group weekly, viatelephone, to monitor their sleep habits.Post-intervention (10 days after the end of the intervention)Ten days after the end of the intervention, the volunteers started again theactigraphy recording, filled the questionnaires and the sleep diary.No follow-up was performed in this study.The general mixed model (GMM) was applied, considering as dependent variables thescores in the questionnaires and the parameters obtained by the actigraphyrecording. The group , gender, time , and the interaction between group and time were considered as fixedfactors, and age and BMI as covariates in the model.27.Individuals were added as a random factor to verify the effect ofintra-individual variability (placebo effect). This choice was made based ontheory  and on the analysis ofcomponents of variance (ICC>10%)p<0.05). The software used for the analyses was Jamoviversion 1.2.27. Effect size was calculated using Lenhard e Lenhard(2016)28calculator.The post hoc test adopted was Bonferroni and the significance level was 5%/apre; and, to describe the percentages of thetreatment effect, these values were multiplied by 100. Negative values indicatehigher values after the intervention and positive ones, the opposite.Treatment effect: the treatment effect was calculated using the formulasdescribed by Herbert (2000)p=0.005; β=3.4 95%CI: 1.0-1.4; ICC=51,d=2.0 - large effect 95%CI: 0.4-3.6) when compared to thecontrol group   .p=0.22ICC=2.7.10-15%), efficiency ,total sleep time , and awake time after sleeponset  (We did not observe a significant effect of the intervention on the sleepparameters obtained from actigraphy: latency (ICC=26%) .p<0.001, β=5.0, 95%CI: 4.4-5.6,ICC=44%, d=6,2 - large effect 95%CI: 3.0-9.3)  .p=0.36, ICC=45%)and on fatigue . For fatigue, a significanteffect of the group factor alone was observed : regardless of the interventionpatients diagnosed with CMT disease had worse fatigue perception than thecontrol group  in pain , which cThe results obtained in this study partially corroborated the proposed hypothesis.The specific physical therapy program improved the subjective sleep quality andnocturnal cramps  and 3 bu31 obtainedsimilar results: improvement in the subjective sleep quality after three and sixmonths of intervention in people with chronic low back pain with a similarintervention protocol and ways of assessing the outcomes  but with some differences mainly in the duration of theintervention (8 weekly meetings vs. 3 weekly meetings performed inour study).Eadie et al. (2013)32.It has also been observed that stretching before bed has been able to improvenighttime cramps by reducing their frequency and intensity. In our study we did notassess frequency, but we obtained similar results, with a reduction in the intensityof nighttime cramps in patients with CMT disease, but it is worth noting importantdifferences in the application of the intervention (time and duration of theintervention)33. However, it is worth noting that there are a wide variety ofoutcomes and intervention protocols, which hinders a more effective match with ourresults33.To our knowledge, this is the first study whose objective was to evaluate the effectof physical therapy on sleep in patients with CMT, so evidence for comparison withour outcome is lacking. Studies indicate that physical therapy is considered auseful tool for symptom management in CMT disease since it can improve the patient’smuscle functionality of patients34 demonstrated that moderate physical exercises are moreeffective in improving sleep quality than intense ones.Despite this subjective improvement in sleep quality, the same was not observed inthe objective measurements obtained from the actigraphy recording . Wang an15,31,34, we did not observe this effect. We attribute this result tothe lifestyle of the volunteers, who are from cities in the interior of Sergipe, andwere able to take daytime naps, which may have impacted their sleepiness.The program also had no effect on daytime sleepiness . Althoug35. This may explain the absence of improvement in actigraphyparameters.The ICC values indicate that our results also showed high intra-individualvariability, which may be an indication of a placebo effect. Studies show that foranalgesia this response may be caused by the expectation of change in symptomscombined with emotional motivations36,37.Sleep is regulated by biological factors (sleep pressure and circadian rhythm) butalso by social and environmental factors, which influence sleep duration.Interestingly, the sleep patterns of the volunteers in our study are like ancestralsleep theories: sleep during the night of 5 to 7 hours and naps distributedthroughout the day38. In this work, we found no significant effect of theprogram on perceived pain and fatigue. One possible factor is that perhaps theduration of the intervention was not enough to lead to these modifications since astudy by Eadie et al. (2013)31observed effects after three and six months of intervention.Data in the literature show that rehabilitation programs that use physical therapy asa foundation  are the most effectivefor the mid-to-long-term treatment of fatigue39,40.It is also worth noting that although the physical therapy program used in this studywas based on the joint strengthening of the lower limbs, there was noregion-specific stimulation or focus on systemic condition. Studies show thatdepending on the type of peripheral pain, stimulation can be done on the skin withdifferent materials to reduce the sensitization of peripheral nervesTherefore, further studies evaluating the effects of physical therapy, adoptingdifferent protocols  may answer this gap in theliterature and offer more conclusive data.A valid limitation to be pointed out is the sample size. We know the importanceof adequate sample size, but due to the characteristics of the disease(uncommon) and the difficulty in adhering to the physical therapy program, wewere not able to fit the sample to the ideal size. Since this is a pilot andpreliminary study, future studies with a more adequate sample size will be ableto better elucidate the relationship between sleep and pain, and the possibleeffects of physical therapy on these parameters.41. We triedto reduce these imbalances by using an age- and sex-matched control group andcontrolling for some variables in the statistical analysis42 but we are aware of thislimitation.Other limitation is the lack of randomization since a non-randomized allocationcan lead to biased interpretation of the data. This does not invalidate ourresults, as non-randomized studies are considered moderate level ofevidenceOur hypothesis was partially corroborated by our results: the specific physicaltherapy program for Charcot-Marie-Tooth was able to improve subjective sleep qualityand nocturnal cramps, but we did not observe an effect on objective sleepparameters, assessed by actigraphy.