Brucella abortus, Leptospira sp., Neospora caninum, leukosis, infectious rhinotracheitis, and BVD using univariate analysis followed by logistic regression. Haplotype 2 of TLR2 was present in 70% of the animals that tested positive for N. caninum infection. Haplotypes of TLR10 and TLR6 and IL10RA were more common in seronegative animals. Haplotypes related to the gene IL10RA were associated with animals negative to all infections. Curraleiro Pé-Duro cattle presented polymorphisms related to resistance to bacterial, viral, and N. caninum infections.Susceptibility to diseases is inherited and can be transmitted between populations. Single-nucleotide polymorphism (SNPs) in genes related to immune response is associated with diseases in cattle. This study investigated SNPs in the genomic region of cytokines in 702 samples of Curraleiro Pé-Duro cattle and associated them with the occurrence of antibodies in brucellosis, leptospirosis, neosporosis, leukosis, infectious bovine rhinotracheitis (IBR), and bovine viral diarrhea (BVD) tests. DNA samples were evaluated by the kompetitive allele-specific polymerase chain reaction (KASP) method to identify polymorphisms. The gametic phase and SNP haplotypes were determined with the help of PHASE 2.1.1 software. Haplotypes were associated with serological results against  Mycobacterium bovis, and other infections [Curraleiro Pé-Duro (CPD) is a local Brazilian bovine breed with historical, cultural, and ecological values, but its population size is small and faces the risk of extinction . The brefections .BOLA), containing a set of closely linked polymorphic genes that code for cell surface proteins essential for the adaptive immune system [TNF) [Disease resistance is determined by many genes, including the ones that encode the regulatory molecules of the immune system. One of the most critical genomic regions involved in disease resistance is the bovine histocompatibility complex, called bovine leukocyte antigen (e system . The comem [TNF) . In the em [TNF) .NRAMP 1) [SLC11A1), immunoglobulin genes, and T-cell receptor genes [SLC11A1 encodes an iron ion transmembrane transport protein, while the toll-like receptor 1 (TLR1) encoding gene is responsible for pathogen-associated molecular pattern-recognition proteins (PAMPs) and regulates cytokine production [The immune response depends of the contributions of multiple genes that produce molecules involved in the regulating the activity of the host immune system. The complex regulation of immunity includes the gene encoding protein 1 (NRAMP 1) , which ior genes . SLC11A1oduction ,10,11. Coduction .TLR), which are present in phagocytic cells, are activated by microbial products and emit intracellular signals that activate cytokine production. The TLR2 receptor is activated by cell wall molecules of Gram-positive bacteria, TLR4 by bacterial membrane lipopolysaccharides, TLR5 by flagellin, and TLR9 by bacterial DNA [The type of pathogen involved in the infection determines the type of immune response. Toll receptors  by antigen-presenting cells stimulates natural killer cells that lyse infected cells [N. caninum infection triggers a cellular response mediated by pro-inflammatory cytokines such as interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and the induction of inducible nitric oxide synthase (iNOS) associated with phagocytic activity [Leptospira sp. modulate the immune response mediated by TLRs and the production of pro-inflammatory cytokines [Bacterial infections, such as those caused by ed cells ,16. N. cactivity . The proytokines .TNF-α. In vitro BVDV infection, in the presence of another pathogen, decreases the production of TNF-α, thereby contributing to the immunosuppression commonly observed in animals infected postnatally. There is a decrease in chemotaxis and the release of an inhibitor of interleukin 1 activity, which enhance the survival of the virus in the host [Bovine leukosis virus (VLB), bovine herpesvirus type 1 (BoHV-1), and bovine viral diarrhea virus (BVDV) induce the production of the host .IFN-γ is induced and promotes cytokine and T-cell response, interfering in immune response to pathogens, such as bovine herpesvirus 1 [The persistent infection in the fetus infected by the bovine viral diarrhea virus (BVDV) noncytopathic (ncp) strain induces higher neutralizing antibodies as compared to homologous cytopathic strain  and estasvirus 1 .Given the genetic role in immune regulation, marker-assisted selection can be a cheap and simple tool to identify the resistant animals based on single nucleotide polymorphism (SNPs) variations involved in the response to pathogens . It is iSingle nucleotide polymorphisms are a substitution of one nucleotide base in DNA sequence, some of which can affect gene transcription and protein activity, promoting differences between species and individuals. SNPs’ markers are used in genetic and genome-wide association studies, because they can be found throughout the genome and are stably transmitted to progeny. Most SNPs are biallelic and have lower information content in comparison to microsatellite markers, so many SNPs are necessary to evaluate differences in population ,25.A combination of alleles occurring on the same autosome or sexual chromosome region, inherited together in a population by the principle of linkage disequilibrium, is termed a haplotype. Each individual has two haplotypes, and one population may have numerous haplotypes. In regions of high linkage disequilibrium, haplotypes that contain, or are correlated to, casual variants can serve to identify disorders or diseases .This paper aims to identify the SNPs’ haplotypes in immune response genes and their association with serology test results in Curraleiro Pé-Duro cattle raised in Brazil.TM spectrophotometer  and standardized at 20 ng/µL, excluding samples that presented OD260/OD280 lower than 1.5 and higher than 2.0.The samples were obtained from the Pró-Centro Oeste Network database, research, and knowledge transfer in Brazil, which maintains blood, serum, and DNA samples as well as epidemiological data of CPD cattle herds. Aliquots of blood samples of animals of both sexes and all ages, contained in the database of samples of the Network at the Universidad de Córdoba, Spain, were subjected to DNA extraction by the salting out method  with somwww.lgcgenomics.com (accessed on 23 February 2017)) in the United Kingdom, where they were processed by the kompetitive allele-specific polymerase chain reaction (KASP) technique.The samples were separated; 940 samples corresponding to 10 plates with two controls in each were sent to the LGC Genomics laboratory  of Federal University of Goiás, protocol Nº 106/19.TLRs, node-like receptors, interleukins, and interferon-γ and were related to tick infestations in cattle [http://www.ensembl.org/Bos_taurus/Info/Index (accessed on 20 June 2016)).The SNPs used  were pren cattle , subclinn cattle , and bovn cattle . The priWith the help of Kbioscience software, the wavelength data emitted by FAM and HEX were plotted on the x- and y-axes, respectively, and ROX values were used as a reference for data normalization, eliminating problems related to the variation in the volume of liquid in each sample. The homozygous genotypes were separated from heterozygous genotypes according to the marked color and position in the graph . The genQuality control of genotype data was performed using the software Excell. Monomorphic SNPs were removed, and SNP markers with a minor-allele frequency (MAF) <1%, and individuals with a call rate <85% were discarded to preserve samples with the best quality of genotyping , leavingGiven the genotypes, the gametic phase of the individuals was determined with the help of PHASE 2.1.1 software to establish the haplotypes. SNPs were grouped according to the chromosomes in which they were present to define the effect of the combination of SNPs on the phenotype. For each chromosome analyzed, the algorithm was applied five times and 100 iterations, a sampling interval, and 100 burn-in through the Markov chain were used ,35. EachIn Brazil, the cattle are mainly raised in pasture systems, which implies an exposure to several pathogenic agents simultaneously. Thus, this study was designed to identify haplotypes associated with resilience in local breeds populations adapted to harsh environments and naturally exposed to infections. For locally adapted breeds from small cattle populations, the limited number of records for quantitative traits, especially for fitness and health traits, is a special challenge in genomic studies.Individuals from 19 different farms were selected for genotyping using a selective genotyping approach. In this regard, the selection criteria were the herd prevalence for the diseases considered in the study, which indicates the herd was naturally exposed to the pathogens.The phenotypes were defined according to the positive or negative reaction to serological tests . The anaB. abortus, Leptospira sp., N. caninum, bovine leukosis virus, bovine herpesvirus type 1 (causing infectious bovine rhinotracheitis), and bovine viral diarrhea (BVD). Serological analyses are a part of the Pró-Centro Oeste Network database and were carried out from 2011 to 2013.The SNPs were associated with a serological reaction (positive or negative) against ®, Curitiba, Brazil) was used against B. abortus, considering the agglutinated samples positive, and the 1:100 cut-off point was used in microagglutination tests against 19 serovars of Leptospira sp.  and indirect immunofluorescence against tachyzoites of N. caninum NC-1 isolate [Serological tests for viral infections included enzyme immunoassay tests , which were performed according to the manufacturer’s cut-off point. The buffered acidified antigen (AAT) method  statistical program using the packages Epitools for univariate analysis and Epicalc and Car for regression. A 5% significance level was considered in the Chi-square and Fisher’s tests ,36.The epidemiological variables obtained from the epidemiological questionnaires of the Pró-Centro Oeste Network were herd size, rearing of other breeds, animal acquisition, quarantine enforcement, use of common pastures or rental of pastures, presence of flooded area, slaughter at the property, occurrence of abortions, vaccination, type of management, presence of rodents, and veterinary assistance. These variables are associated with the risk of infection; hence, some animals, even in the presence of an environment favorable for infections, may not exhibit the infection. The significant variables were added to the multivariate generalized linear model using binary logistic regression. For this purpose, the glm function of the R software was utilized .In the quality control stage, two monomorphic SNPs, rs207532826 and rs42395524, were removed from the analyses. Based on the call rate criterion, 238 samples with >15% failures were excluded, and only 702 of the 940 initial samples were left. SNP quality control prevents newly pinned markers and samples with low informative content from generating false results in association tests.B. abortus, 42.07% (292/694) for Leptospira spp., 36.87% (257/697) for N. caninum, 21.07% (146/693) for VLB, 64.65% (450/696) for BoHV-1, and 40.38% (279/691) for BVD. The frequencies of each infection by property (owner) were calculated by considering only the samples used in the evaluation of SNPs (The frequencies of seropositive reactions were: 0.85% (6/702) for  of SNPs .The low prevalence of B. abortus agglutinins expresses the effective control of disease by the National Program for the Control and Eradication of Animal Brucellosis and Tuberculosis and the correct vaccination of heifers against brucellosis. On the contrary, there is no vaccine available against N. caninum infection in cattle, and the control strategies are not suitable.To assess whether SNPs are in linkage disequilibrium, the markers were grouped according to the chromosome, and haplotypes representing different genotypes were formed . SNP rs4The frequencies of infections were tested for each polymorphism by univariate analysis using Fisher’s test, and the significant differences are presented in p < 0.05  identification of the epidemiological variables that represent risk factors for the onset of diseases through the Chi-square test. The non-significant variables were evaluated as having little relevance for increasing the incidence of the disease; (2) identification of haplotypes associated with diseases. Logistic regression was performed considering haplotypes and significant epidemiological variables as independent variables in the model. The final regression results are expressed in Logistic regression was used to observe whether the response variable (presence/absence of antibodies) was related to the haplotypes of each chromosome and related to epidemiological variables.When the epidemiological factors were included into ANOVA analysis , the conB. abortus animals. For Neospora sp. infections, we found c10_h7 haplotype in 25.29% (130/514) positive animals, c17_h2 haplotype present in nine negative and 21 positive animals. Haplotype c18_h1 present in 104 positive and 130 negative animals. In bovine leucosis infection, the haplotype c27_h2 was present in 31.5% (46/146) positive animals.Haplotypes c8_h1 and c15_h2 were present in five of six positive It was possible to identify potential gene loci candidates for bovine resistance to infectious diseases in the 10 chromosomes studied. The BTA15 chromosome haplotypes were significant against the infections defined. Of the five haplotypes described, haplotypes 1 and 2 (c15_h1 and c15_h2) were more frequent in seronegative animals. Haplotypes 1, 5, and 6 in BTA6 were more common in animals negative against leptospirosis, neosporosis, viral diarrhea, and leukosis.p < 0.05) associated with positivity or negativity for the infection. Haplotypes were generally more frequent in seronegative animals and were, therefore, associated with the absence of infection.Haplotype blocks containing two, three, and four SNPs were found that were significantly  and in families capable of recognizing bacteria, protozoa, and fungi [Polymorphisms in nd fungi ,38.TLR10 are found in the placental tissues and are involved in protecting the fetus. TLR10 functions as a co-receptor of TLR2, a peptidoglycan agonist of Gram-positive bacteria that acts by inducing cell apoptosis and a reduction in chemokine secretion [TLR10 have a greater ability to fight infections and reduce the transmission of pathogens than those that do not express the receptor.The pathogens investigated are sexually transmitted and are important causes of reproductive loss in cattle. In humans, the proteins expressed by ecretion . AnimalsIL10RA were associated with all infections. The SNPs selected in BTA15 are present in the gene of interleukin 10 (IL-10) subunit α receptor. IL-10 is a cytokine produced by CD4+ T-cells, B-cells, and macrophages [IL-10 has immunoregulatory potential and inhibits pro-inflammatory cytokines, mainly FNT, IL-1, and IL-6, produced by activated macrophages and monocytes [IL-10 reduces inflammation during infections, preventing the onset of deleterious lesions. However, it can also promote the persistence of pathogens by interfering with the immune response, such as the persistence of Leptospira spp. in the kidneys of reservoir animals [The SNPs examined in rophages . IL-10 honocytes . Further animals .IL10RA and seronegative animals may be due to the regulatory effect of IL-10. In Leptospira spp. infection, IL-10 increases expression in the early post-infection days in resistant rats, while pro-inflammatory cytokine levels decrease [IL-10 during leptospirosis is to mitigate the deleterious effects caused by the increase in pro-inflammatory cytokines, such as interleukin 1 β (IL-1β), which is related to organ failure in infected hosts. On the other hand, it exerts an inhibitory effect on bacterial clearance, causing bacteria to remain in the kidneys and be eliminated by the host [The association between haplotypes in decrease . The immthe host .TLR8 are found in this chromosome. TLR8 recognizes the invasion of viruses and induces the innate immune response; hence, it is often associated with BVD and type 3 parainfluenza caused by the same bovine herpesvirus type 1, resulting in infectious rhinotracheitis as well as other viral diseases. Because these diseases are disseminated, animals are likely to be exposed for a long time. Furthermore, owing to evolutionary pressure, the pathogen–environment interaction of ancestral populations may have generated patterns of variation within the TLR3 and TLR8 genes that are seen as selection signatures [The X chromosome was associated with neosporosis from the Chi-square test but was not associated with any pathogen in the regression analysis. Gene-encoding gnatures .The SNP rs29026690, in the current study associated with BVD, was described in an investigation of the broad association of the genome with an increased proviral load of the bovine leukosis virus in Japanese Black herds (odds ratio 2.745), within a locus of independent quantitative characteristics (QTL—quantitative trait loci) on chromosome 23. In the same study, they identified a minor association of the disease with SNP rs17872126 (odds ratio 0.414) .IL10RA and TLR3 were present in a high rate of seronegative animals and may indicate a possible genotype of resistance to bovine leukosis virus. On the other hand, 70% of animals positive for N. caninum infection had haplotype 2 from TLR2.In the analysis of haplotypes associated with infections, significant differences were identified in chromosomes 5, 6, 8, 10, 15, 17, 18, 27, and X. Haplotypes from IFNG gene, which encodes γ interferons playing an important role in the innate host response. In this study, alleles and haplotypes were significant only when not associated with other explanatory variables. The non-association between haplotypes and epidemiological characteristics in response to infection observed in this study does not denote the lack of association. The conclusions obtained from case-control epidemiological studies are limited, and differences in the susceptibility of animals could be due to several factors such as errors in the classification of individuals in the categories of case (positive) and control (negative) [The herd and breed of animals were significant variables in a study of the association of paratuberculosis infection and SNPs in the region of the bovine egative) , as obseEven though other studies have shown the possibility of significant results between markers and characteristics, its occurrence in populations may not be replicable. If the same locus of quantitative characteristics is segregating into different populations, the results will still be different, because the allelic frequencies for each marker and the mutations are different. Genetic resilience can present various phenotypes, such as exposed individuals who do not develop an infection, exposed subjects who become asymptomatic carriers, and individuals who develop clinical signs of the disease but manage to cure themselves ,45.Alleles that confer resistance in one breed need to be validated in other breeds. It was observed that the alleles associated with a lower load of tick infestation in one population do not perform the same function in others. Genetic markers or haplotypes do not show precise effects between different breeds , indicatAnimals of local breeds have proven resistance to infections; however, the selection for resistance characteristics is still at an early stage. CPD animals can be immunologically challenged to investigate the type and magnitude of the immune response in those carrying the haplotypes identified as resistance markers. Animals carrying resistance alleles must be included in breeding programs, and their genetic material should be preserved in the germplasm bank.It is known that cattle production characteristics have been widely researched owing to the direct economic return. However, investments in health and the development of biotechnology make it feasible to control diseases via molecular markers as a way of supporting the sanitary strategies that have already been employed.TLR2 was present in 70% of animals positive for N. caninum infection, while haplotypes related to IL-10 and TLR10 were common in seronegative animals. The epidemiological variables demonstrate significance with serological status. Further research in experimentally infected CPD animals in a controlled environment should be done to validate the association between the presented SNPs and diseases antibody levels.SNP-type markers related to the risk of infectious diseases were present in animals of the CPD breed. The polymorphisms formed haplotype blocks for each chromosome, and chromosomes 5, 6, 8, 10, 15, 17, 18, 27, and X were associated with diseases. Haplotype 2 of


An auto-DODS can also supply pulsed-flow oxygen when it detects apnea, whereas a conventional DODS has only standard sensitivity. This randomized, open-label, crossover pilot study compared the performance of an auto-DODS with that of a conventional DODS.: When using portable oxygen, a demand oxygen delivery system (DODS), which senses the beginning of inhalation and delivers a bolus of oxygen, is often used. However, conventional DODS may not supply sufficient oxygen when reduced tidal flow fails to trigger the flow sensor. Recently, “auto-DODS,” which detects the negative pressure of inhalation and switches among 3 trigger sensitivity levels (: We recruited patients with chronic obstructive pulmonary disease (COPD) or interstitial pneumonia receiving long-term oxygen therapy. Interventions were performed on 2 different days for each participant. On each day, an auto-DODS or a conventional DODS were tested at rest for 30 minutes and during the 6-minute walk test. The primary outcome was mean oxygen saturation (SpO2). Secondary outcomes were the ratios of time for each sensitivity level and pulsed-flow oxygen when using the auto-DODS, total time desaturated below SpO2 90%, percentage of time desaturated below SpO2 90%, minimum SpO2, mean and maximum pulse rate, six-minute walk distance, recovery time after 6-minute walk test, modified Borg scale, comfort, and discomfort index.high or extra high sensitivity level was observed in addition to standard sensitivity in 6 of 8 participants. During the 6-minute walk test, only standard sensitivity was observed in 6 participants. Mean SpO2 differences between the auto-DODS and conventional DODS at rest and during the 6-minute walk test were −0.6  and 0.0  , respectively, neither of which were significant (P = .73 and P = .99). There were no significant differences in secondary outcomes. There were no adverse events when using the auto-DODS.: When using the auto-DODS at rest, a : This study showed that the auto-DODS did not show superiority in oxygenation either at rest or during exercise compared to a conventional DODS. The auto-DODS was shown to supply oxygen safely and detect inhalations with various trigger sensitivities. It has already been proven that long-term oxygen therapy can reverse the progression of pulmonary hypertension and improve the quality of life and prognosis of patients with COPD. It is still not clear whether the use of portable oxygen itself improves the prognosis of hypoxemic patients, but it has already been shown that portable oxygen helps to relieve the symptoms of breathlessness during exertion, improve mobility and increase compliance with oxygen therapy.–6Portable oxygen is also prescribed to an increasing number of hypoxemic patients.[3] However, there is a concern that a DODS may not supply sufficient oxygen because reduced tidal flow may fail to trigger the flow sensor.When using portable oxygen, a demand oxygen delivery system (DODS) that senses the beginning of inhalation and immediately delivers a short bolus of oxygen  is often used because this system limits oxygen consumption and extends the time that patients can use an oxygen cylinder in an ambulatory setting.standard, high, and extra high), has been developed recently to improve the efficacy of oxygenation in hypoxemic patients. This oxygen concentrator can also supply pulsed-flow oxygen when it detects apnea. Thus, we hypothesized that this auto-DODS is superior to a conventional DODS in improving oxygenation, and the present study aims to compare the performance of an auto-DODS with that of a conventional DODS both at rest and during exertion.To solve this problem, a new type of portable oxygen concentrator with an “auto-DODS,” which can detect the negative pressure of inhalation and switch among 3 trigger sensitivities . This trial was also registered in the Japan Registry of Clinical Trials (jRCT) on August 23, 2019 . The registration period was set from August 2019 to April 2020. The follow-up period was 2 months from the date when the final case was registered. The study protocol was reported by Nagano et al.Written informed consent was obtained from all the participants included in this study. All procedures performed were in accordance with the ethical standards of the institutional and national committees and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.2.2 or mentioned in the jRCT .We recruited patients aged 20 years or older with COPD or interstitial pneumonia who received long-term oxygen therapy (less than 4 L/min during the daytime). Detailed inclusion and exclusion criteria are summarized in a previous paper of this study protocol,2.3Interventions were performed on 2 different days for each participant: the first day of testing (Day 1), at a 4-week interval, and on another day of testing (Day 29). On Day 1 and Day 29, the auto-DODS or conventional DODS were tested both at rest for 30 minutes and during the 6-minute walk test. The oxygen flow rate during these tests was adjusted based on the prescription flow rate in participants’ daily lives, and the flow rate was set to be the same for both the auto-DODS and conventional DODS.After registration, participants were randomly assigned to either Group 1  or Group 2  . A detailed study protocol that describes randomization and masking procedures has been mentioned by Nagano et al.2.4Because no prior study has been conducted in hypoxemic patients using an auto-DODS, it was not possible to statistically ascertain the required number of participants in this trial. However, the sample size was determined to be 26 cases (13 cases per group) in consideration of feasibility within the study period. Because this is a pilot study, we decided that the registration period won’t be extended even if the sample size was not fulfilled during the period.2.5standard, high, and extra high: the higher sensitivity level can detect shallower inhalation), and thereupon directs the solenoid valve to open momentarily to deliver a bolus of oxygen. Specifically, the negative pressure of inhalation detected by high sensitivity is half that of the pressure detected by standard sensitivity, while the pressure detected by extra high sensitivity is one tenth of the pressure detected by standard sensitivity. With higher sensitivity level, oxygen can be supplied even during shallower inhalation. The trigger sensitivity is adjusted automatically; when the valve opens fewer than a predetermined number of times during a fixed interval, the sensitivity increases by 1 step, and when the valve opens more than the predetermined number of times, the sensitivity decreases by 1 step. Additionally, the auto-DODS supplies pulsed-flow oxygen when it detects apnea. The conventional DODS senses the negative pressure of inhalation with standard sensitivity only, and it does not respond to apnea .On Day 1 and Day 29, a Hi-Sanso Portable αII oxygen concentrator equipped with auto-DODS and conventional DODS settings  was used. Through a nasal cannula, the auto-DODS senses the negative pressure of inhalation and switches its trigger sensitivity among 3 levels  and pulse rate were continuously recorded for 30 minutes by a pulse oximeter, PULSOX-Me300 . During the 6-minute standardized walk test, participants were also equipped with either of the DODS, and both SpO2 and pulse rate were similarly recorded. Apnea alarms were also recorded during the measurement time.2.6The primary outcomes were mean SpO2 at rest and during the 6-minute walk test. The secondary outcomes were1.the ratios of time for each sensitivity and pulsed-flow oxygen when using the auto-DODS (percentage of equipment time spent on each sensitivity level),2.total time desaturated below SpO2 90% at rest and during the 6-minute walk test,3.percentage of time desaturated below SpO2 90% at rest and during the 6-minute walk test,4.minimum value of SpO2 at rest and during the 6-minute walk test,5.mean pulse rate at rest and during the 6-minute walk test,6.maximum pulse rate at rest and during the 6-minute walk test,7.six-minute walk distance,8.recovery time ,9.modified Borg scale ,10.comfort when using the DODS measured by a numerical rating scale  and11.discomfort index when using DODS measured by questionnaires .The details of the discomfort index questionnaires are shown in the Supplemental Digital Content.2.7P values were calculated based on a t-test. Additional details were described by Nagano et al. In all cases, P values ≤.05 were considered significant. Analyzes were carried out using SAS Proprietary Software 9.4, SAS/STAT 15.1, and JMP 9.0.2 . Statistical analyses were performed by a biostatistician at the Clinical and Translational Research Center at Kobe University Hospital (S.M.).Data are summarized using the median (range) or mean (standard deviation) for continuous variables and numbers for categorical variables. The differences in outcomes between the auto-DODS and conventional DODS and their 95% confidence intervals (CIs) were also calculated. 33.1During the registration period, 8 participants were included in this trial. The flow chart of participant randomization is shown in Figure 3.21% predicted, FVC % predicted, oxygen flow at rest, and oxygen flow during exertion).The characteristics of the participants in this study are summarized in Table 3.3standard, high, and extra high) and pulsed-flow oxygen when using the auto-DODS are shown in Figure The ratios of time at each sensitivity level . There were also no significant differences between the auto-DODS and conventional DODS in any of the secondary outcomes.As mentioned above in Fig. , in Grouextra high and high sensitivity accounted for 75.4% and 24.6% of the measurement time, respectively , he had chronic heart failure as an underlying illness. In this participant, when using the auto-DODS, 3.5P = .99). There were also no significant differences between the auto-DODS and conventional DODS in any of the secondary outcomes.Table 3.6There were no adverse events or problems when using the auto-DODS and conventional DODS in this trial.4 Moreover, the Japanese respiratory society reported that among patients using home oxygen therapy, more than 70% of them are using the DODS in an ambulatory setting.Home oxygen therapy is an established treatment for hypoxemic patients. In recent years, an increasing number of patients with chronic respiratory diseases, such as COPD, interstitial pneumonia, and lung cancer, have been receiving home oxygen therapy. According to this concept, fast space fills during the earliest part of inhalation and is well perfused with pulmonary circulation. On the other hand, slow space is filled at the very end of inhalation and is poorly perfused. Based on this concept, it is thought that providing oxygen flow at the onset of inhalation by the DODS leads to more efficient arterial oxygenation. Moreover, a majority of the respiratory cycle is spent in exhalation, while most of the time during inhalation is spent filling the dead space. Therefore, the concept of the DODS is very reasonable.There is a concept of “fast space ventilation” and “slow space ventilation”. However, in another study, Roberts et al showed that among subjects with severe COPD whose oxygen saturation fell below 90% during exercise, the DODS was less effective for oxygenation during walking than continuous oxygen flow. We thought that reduced tidal flow or rapid respiratory rates during rest or exercise could fail to trigger the flow sensor when using the DODS. Therefore, in this trial, we tested whether the auto-DODS, which can detect the negative pressure of inhalation and switch among 3 trigger sensitivity levels , could improve oxygenation compared to the conventional DODS, which senses the negative pressure of inhalation with standard sensitivity only.In a previous study, it was reported that the DODS can maintain arterial oxygenation equivalent to continuous oxygen flow both at rest and during exercise in hypoxemic patients with COPD.standard sensitivity, high or extra high sensitivity was observed in 6 of the 8 participants when using the auto-DODS. However, in contrast to our hypothesis, there were no significant differences between the auto-DODS and conventional DODS in mean SpO2 or any of the other variables. The estimates indicated as differences between the 2 DODSs in mean SpO2, considering their 95% CI, were approximately zero, as shown in Table extra high and high) with the auto-DODS could prevent trigger failure that would have happened with the conventional DODS. Thus, among patients with shallow breathing at rest, the auto-DODS may show superiority for improving oxygenation compared to the conventional DODS, and this should be elucidated in future studies.At rest, in addition to standard sensitivity was observed in 6 out of 8 participants when using the auto-DODS. This means that flow sensors can be successfully triggered during exercise in most of the participants by using only standard sensitivity and that the auto-DODS is unlikely to have an advantage in triggering flow sensors over the conventional DODS during exercise. Additionally, during the 6-minute walk test, no significant difference was observed in SpO2 between the auto-DODS and conventional DODS.During the 6-minute walk test, only The limitations of the current clinical trial are as follows. First, the number of participants was small (8 participants). Although it was not possible to statistically determine the required number of participants because of the exploratory design of this trial, further studies with more subjects are needed to confirm our findings. Second, oxygenation of the participants was assessed not by arterial oxygen saturation or arterial oxygen pressure but by SpO2 measured by a pulse oximeter. However, pulse oximetry has been proven to be effective in measuring arterial desaturation, and we think that this limitation has no serious effect on the results of this study.In conclusion, this clinical trial has shown that the utility of the auto-DODS in maintaining oxygenation at rest and during exercise was equal to that of the conventional DODS among patients with chronic respiratory failure. Additionally, the auto-DODS was demonstrated to be safe when used for hypoxemic patients both at rest and during exercise.The authors thank the staff of the Clinical and Translational Research Center of Kobe University Hospital for their support in this trial. The authors also thank the staff of the Division of Respiratory Medicine of Kobe University Hospital for recruiting and caring for patients in this trial. The authors also thank Springer Nature Author Services for editing our manuscript for proper English language.Conceptualization: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Daisuke Hazama, Naoko Katsurada, Masatsugu Yamamoto, Motoko Tachihara, Yoshihiro Nishimura, Kazuyuki Kobayashi.Data curation: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori.Formal analysis: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori.Funding acquisition: Tatsuya Nagano, Yoshihiro Nishimura.Investigation: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori, Daisuke Hazama, Naoko Katsurada, Masatsugu Yamamoto, Motoko Tachihara, Yoshihiro Nishimura, Kazuyuki Kobayashi.Methodology: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori, Yoshihiro Nishimura.Project administration: Tatsuya Nagano, Yoshihiro Nishimura.Resources: Tatsuya Nagano.Software: Tatsuya Nagano, Sae Murakami, Takashi Omori.Supervision: Tatsuya Nagano, Sae Murakami, Takashi Omori, Yoshihiro Nishimura, Kazuyuki Kobayashi.Validation: Takehiro Otoshi, Tatsuya Nagano.Visualization: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori.Writing – original draft: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori.Writing – review & editing: Takehiro Otoshi, Tatsuya Nagano, Sae Murakami, Takashi Omori, Daisuke Hazama, Naoko Katsurada, Masatsugu Yamamoto, Motoko Tachihara, Yoshihiro Nishimura, Kazuyuki Kobayashi.


Oxaliplatin (OXA), a third-generation platinum derivative, has become one of the main chemotherapeutic drugs for colorectal cancer and other cancers, but reports of adverse reactions are also increasing with the extensive application of OXA. In this study, post-marketing surveillance was carried out to investigate the safety profile of OXA in a real-world setting in Chinese cancer patients to provide a reference for the rational application of OXA.All patients with cancer who received OXA-based chemotherapy in 10 tertiary hospitals in Hubei Province, China, between May 2016 and November 2016 were enrolled. A central registration method was used to document patients’ demographics, clinical use, and any incidence of adverse reactions to OXA. All adverse drug reactions (ADRs) were collected and analyzed to assess causality, severity, treatment, and outcome.In total, 3687 patients were enrolled in this study. Approximately 64.6% of the patients were male, and 68.8% were aged 50-70 years, with a mean age of 55.3 years. The proportions of patients diagnosed with colorectal and gastric cancers were 59.3% and 31.6%, respectively. In this study, the overall incidence of ADRs and serious ADRs was 42.7% and 1.3%, respectively. The most common ADRs were gastrointestinal disorders (25.7%), blood disorders (21.1%), and peripheral nervous system disorders (8.0%). The serious ADRs identified were hypersensitivity reactions, thrombocytopenia, abnormal hepatic function, and leukopenia/neutropenia. The median onset of gastrointestinal toxicity, myelosuppression, peripheral neurotoxicity, and abnormal hepatic function was 1 d, 5 d, 1 d, and 14 d, respectively. The majority (84.7%) of hypersensitivity reactions were mild to moderate, and the median time to onset of these reactions was within the first 20 min of OXA infusion. Almost 88.0% of patients who experienced ADRs recovered or improved with treatment.Our data suggest that OXA-induced ADRs are very common in Chinese patients with cancer; however, more attention should be paid to hypersensitivity reactions caused by OXA. This study provides a valuable reference regarding the safe application of OXA in a real-world setting. Oxaliplatin (OXA) is a third-generation platinum-based anti-tumor drug that inhibits DNA and protein synthesis by forming intra- and inter-strand DNA platinum adducts, leading to tumor growth inhibition and apoptosis . OXA hasWith the extensive clinical application of OXA, adverse reactions towards OXA have also been reported in recent years, mainly including gastrointestinal side effects, hematologic toxicities, and dose-limiting peripheral neurotoxicity –12. MoreThis multicenter observational study was conducted in 10 tertiary hospitals in Hubei Province, China, between May 2016 and November 2016. All patients with malignant tumors treated with OXA were enrolled in this study. All patients were prospectively registered upon initiation of OXA treatment by documenting the patients’ demographics, clinical application, and adverse reactions of OXA using a central monitor method.The investigators received unified and standardized project training according to research work manual and case report forms (CRF)  during ward round and patient education as well as lab results  from HIS system, and the frequency was usually 3 times a week. The investigators needed to report all adverse reactions that occurred after OXA treatment, concomitant medications used, time to onset, symptoms, administered treatments, and outcomes. All reported ADRs were recorded according to the Provisions of Adverse Drug Reaction Reporting and Monitoring with the clinical pharmacists’ assessment of causality. This adverse reaction can be submitted only when the ADR correlation evaluation with OXA is possible or above. The severity of OXA-induced ADRs was classified according to the National Cancer Institute Common Criteria (NCI-CTCAE v5.0), and grade 3 and above adverse reactions were defined as serious ADRs. ADRs were classified using the preferred terms and system organ classes from the World Health Organization (WHO) Glossary of Adverse Drug Reactions. Additional information, such as the results of laboratory tests, was also recorded for serious ADRs.More than 3000 patients receiving OXA were enrolled to detect an ADR in one out of every 1000 patients with a probability of ≥95%. The collected data were verified and encoded by special personnel. All data analyses were performed using SPSS version 24.0 . Descriptive analysis was used to evaluate patients’ demographics (such as sex and age), clinical characteristics such as Karnofsky performance status (KPS), diagnosis, clinical use of OXA, and the incidence of adverse reactions, time to onset, symptoms, administered treatments, and outcomes.A total of 3775 case report forms were enrolled between May 1, 2016, and November 31, 2016. Eighty-eight reports were excluded due to duplication; thus, 3687 patients were enrolled for this safety study. Of the 3687 patients, ADRs were reported in 1575 patients, giving an incidence rate of 42.7%. Most ADRs were grade 1 and 2, with grade 1 ADR reported in 562 patients (15.2%) and grade 2 ADR reported in 965 patients (26.2%). Serious ADRs (grade≥3) were reported in 48 patients (1.3%), showed in The most commonly reported ADRs fell into the general categories of gastrointestinal disorders (25.7%), blood disorders (21.1%), and peripheral nervous system disorders (8.0%) Table 2.The OXA-linked ADRs with the highest frequency were nausea (20.3%), leukopenia (17.3%), neutropenia (12.1%), vomiting (9.8%), anemia (7.0%), thrombocytopenia (5.6%), and peripheral paresthesia or dysesthesia of hands and feet (5.7%). Meanwhile, the most frequent serious ADRs recorded were hypersensitivity reactions, leukopenia, neutropenia, thrombocytopenia, abnormal hepatic function, and vomiting  Table 3.The median time from the start of OXA administration to the occurrence of gastrointestinal disorders, blood system disorders, peripheral nervous system disorders, and hepatic function abnormalities were 1 d, 5 d, 1 d, and 14 d, respectively. For hypersensitivity reactions, the median time from start to occurrence was much shorter, at only 20 min. In addition, the above 5 ADRs occurred in a median cycle of OXA chemotherapy of 3, 4, 4, 4 and 6, respectively. More than 90% of those who experienced gastrointestinal toxicity, myelosuppression, and peripheral neurotoxicity continued OXA therapy. Of those that experienced hypersensitivity reactions, approximately 85% suspended OXA administration and received the corresponding treatment Table 4.The manifestation of OXA-related HSRs is shown in via discontinuation of treatment with OXA and the administration of hypersensitivity treatments, including dexamethasone (61.0%), histamine-receptor 1 antagonist (51.7%), oxygen (28.8%), and epinephrine (8.5%). All patients recovered or improved after the corresponding symptomatic treatment  of enrolled patients were male, within the age bracket of 50 to 69 years old. Epidemiology shows that the incidence rate of colorectal and gastric cancer in males is higher than in females, and most of these occur in middle-aged and older people . Thus, tMoreover, this study showed that patients receiving OXA were mainly diagnosed with colorectal and gastric cancer , and FOLFOX was the most commonly used chemotherapy regimen (61.7%), followed by XELOX 19.5%) and SOX (7.5%). According to NCCN guidelines of colorectal and gastric cancer , 19, FOL.5% and SRegarding the safety results, the overall incidence rate of ADRs was 42.7%, and that of serious ADRs was 1.3%. The most common reported ADRs of OXA in this study were gastrointestinal disorders (25.7%), blood disorders 21.1%), and peripheral nervous system disorders (8.0%), which were consistent with the results of other studies , 20, 21..1%, and Nausea, vomiting, and diarrhea were the most common gastrointestinal side effects of OXA, with a median time of onset of 1.6 d, and most patients recovered or improved quickly. Nausea and vomiting are usually mild to moderate and are readily controlled with the prophylactic administration of standard antiemetics such as dexamethasone or 5-HT3 receptor antagonists . Grade 1In general, hematological side effects caused by OXA include leukopenia, neutropenia, thrombocytopenia, and anemia . In our 2  was reported in 266 patients (8.0%), mainly including paresthesias or dysesthesias of the hands and feet. Most reported OIPN cases were mild, and 96.6% of patients recovered or improved, with the median time to onset of 1 d. OXA induces two clinically distinct forms of peripheral neuropathy. The first is acute OIPN, which is transient, appearing only during or shortly after infusion of OXA, and can be triggered by cold stimulation. The other form, chronic OIPN, is associated with a cumulative dose of OXA and appears after administration of OXA at a total dose of 540-850 mg/m2 . Acute O2 . Thus, oWhile not as common as the previously discussed ADRs, hepatotoxicity also represents a serious ADR that warrants further investigation. There is evidence that the most common type of hepatotoxicity associated with OXA administration is hepatic sinusoidal injury. It is histologically characterized by sinusoidal dilatation, hepatocyte necrosis, and obliteration of hepatic venules due to sinusoidal endothelial cell damage –32. OXA-OXA-related HSRs are another major problem associated with the extensive use of the drug, the occurrence of which may lead to therapy delay, discontinuation of treatment, and even death . The repPrevious studies have noted that OXA-induced hypersensitivity reactions usually occur within the first 30 min of infusion , 47. In However, this study can still be further expanded. First, this was a non-interventional observational study that aimed to observe the safety profile of OXA with no control group, and there was a short observation period of only 6 months and lack of follow-up. Hence, the OXA efficacy in these patients was unknown, and some delayed ADRs such as chronic OIPN may not be immediately reported and were underestimated. Second, patients treated with OXA was the only inclusion criteria implemented, and a comparison of safety accounting for the effects of different diseases and treatment regimens was not performed. In addition, although the researchers have identified OXA-related ADRs, the effects of other drugs cannot be completely excluded because OXA is often used in combination with other chemotherapeutic drugs. Further investigation is also required to identify the risk factors that affect the incidence of OXA-induced ADRs to provide a reference for the rational application of OXA.In conclusion, this large post-marketing surveillance study conducted in more than 3000 Chinese patients preliminarily explored the incidence, characteristics, cycle, occurrence time and outcome of OXA induced ADRs. Overall, OXA induced adverse reactions were very prevalent. Our results showed that gastrointestinal toxicity, hematotoxicity, peripheral neurotoxicity, HSRs and abnormal liver function were the main common ADRs of OXA, in which the latter two had unique characteristics, need more attention, and warrant close monitoring during OXA infusion. Although further studies are still required, this study provides valuable reference for the rational use of OXA and has great guidance for the management of OXA-induced ADRs in routine clinical practice.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by the Ethics Committee of Tongji Medical College, Huazhong University of Science and Technology (No.TJIRB20160504). The patients/participants provided their written informed consent to participate in this study.ZY and RH, first authors, contributed to the data analysis and writing of manuscript. LZ and XY contributed to organization, management, and supervision of the project. XW and XS contributed to data collection, data curation, analysis, and validation. WL and ML contributed to contributed to the editing and submission of the article. CZ and DL, the corresponding authors, involved in conception of the project, performing data analysis, review of manuscript and provision of feedback and comments to first author. All authors contributed to the article and approved the submitted version.This work was supported by the National Natural Science Foundation of China (No: 7187040708), Hubei Center for Adverse drug reaction Monitoring (No: 20160422), Funding for research-oriented clinician plan of Tongji Medical College, Huazhong University of Science and Technology (No: 5001540076); Clinical toxicology foundation of Chinese Society of Toxicology (CST2020CT107).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Feline chronic gingivostomatitis (FCGS) is a common pathology in cats, related to an aberrant immune response. The cause of FCGS remains elusive, despite extensive investigations. A multitude of conditions and infectious agents have been related, without proof of causation, as follows: virus, bacteria, environmental stress, hypersensitivity, etc. In recent years, therapies based on feline adipose-derived mesenchymal stem cells (fAd-MSC) have become an interesting alternative for the treatment of different complex pathologies in cats. Mesenchymal stem cells secrete a wide variety of therapeutic elements, such as bioactive molecules and extracellular vesicles, such as exosomes. It is essential to characterize these elements, to better understand their mechanisms of action. In this study, we show, for the first time, that the proteomic profile of fAd-MSC-derived exosomes, from calicivirus-positive patients with severe FCGS, is altered. Using bioinformatic tools, we have demonstrated the existence of different proteins in the exosomes from diseased patients, responsible for an altered biological effect. In addition, the exosomes do not only experience changes in their cargo, but are also produced in larger quantities. This study might contribute to the better prediction of the clinical outcomes of mesenchymal stem cell treatments in veterinary patients with immune-mediated diseases, such as FCGS.Feline chronic gingivostomatitis (FCGS) is a pathology with a complicated therapeutic approach and with a prevalence between 0.7 and 12%. Although the etiology of the disease is diverse, feline calicivirus infection is known to be a predisposing factor. To date, the available treatment helps in controlling the disease, but cannot always provide a cure, which leads to a high percentage of refractory animals. Mesenchymal stem cells (MSCs) play a pivotal role in the homeostasis and reparation of different tissues and have the ability to modulate the immune system responses. This ability is, in part, due to the capacity of exosomes to play a part in intercellular cell communication. However, the precise role of MSC-derived exosomes and their alterations in immunocompromised pathologies remains unknown, especially in veterinary patients. The goal of this work was to analyze the proteomic profile of feline adipose tissue-derived MSCs (fAd-MSCs) from calicivirus-positive FCGS patients, and to detect possible modifications of the exosomal cargo, to gain better knowledge of the disease’s etiopathogenesis. Using high-resolution mass spectrometry and functional enrichment analysis with Gene Ontology, exosomes isolated from the fAd-MSCs of five healthy cats and five calicivirus-positive FCGS patients, were pooled and compared. The results showed that the fAd-MSCs from cats suffering from FCGS not only had a higher exosome production, but also their exosomes showed significant alterations in their proteomic profile. Eight proteins were exclusively found in the exosomes from the FCGS group, and five proteins could only be found in the exosomes from the healthy cats. When comparing the exosomal cargo between the two groups, significant upregulation of 17 and downregulation of 13 proteins were detected in the FCGS group compared to the control group. These findings shed light on new perspectives on the roles of MSCs and their relation to this disease, which may help in identifying new therapeutic targets and selecting specific biomarkers. Feline chronic gingivostomatitis (FCGS) is an oral mucosal inflammatory pathology, with an estimated prevalence between 0.7 and 12% in the cat population, and a complicated therapeutic approach ,4,5,6. TMesenchymal stem cells (MSCs) are known to play an important role in the homeostasis and repair of numerous tissues, as well as the ability to modulate the responses of both the innate and the adaptative immune system ,10. As tRecently, the exosomes have become particularly interesting, because of their potential diagnostic and therapeutic applications . On one In the last years, it has been demonstrated that the MSCs from human patients with certain immune-mediated diseases are dysfunctional, leading to a better knowledge of these cells and their involvement in diseases ,23,24,25The main objective of this study was to analyze the proteomic profile of fAd-MSC-derived exosomes from FCGS patients who tested positive for calicivirus infection, and to evaluate the existence of alterations in the exosomal cargo. These data will help us to better understand the etiopathogenesis of this disease.All the used protocols received approval by the Ethics Committee of the Andalusian Center for Nanomedicine and Biotechnology . The procedures affecting animals were performed by veterinarians after obtaining informed consent from the owners, which approved the use of biological samples from their cats for this study.Ten European shorthair cats were included in this study . The inclusion criteria for the FCGS group were as follows: severe FCGS confirmed by a biopsy, with all molars extracted at least 6 months earlier and being refractory to immunomodulatory therapies, not showing any clinical improvement . None ofThe cats with FCGS were selected after being subjected to a clinical exploration and hematological and biochemical tests to discard any other existing pathologies. Hyperproteinemia, hyperglobulinemia, and neutrophilia were accepted, as they have been reported to be related to FCGS. The absence of dental roots was confirmed by dental radiographs . The selThe healthy control cats did not have any known pathology. They were subjected to the same clinical and laboratory tests and were proved negative for the aforementioned viral infections.No medication was given to the cats for at least ten days prior to the extraction of adipose tissue. Only analgesic drugs were allowed for the FCGS patients for palliative purposes.A subcutaneous adipose tissue sample (~5 g) was extracted from the inguinal area of each cat after sedation for other procedures not related to this study . The cats were sedated with ketamine , midazolam , and butorphanol . The obtained samples were maintained in Dulbecco’s modified Eagle medium  at 4 °C until taken to the laboratory on the same day.2, 95% RH) and subcultured when reaching >80% confluency. All experiments were performed with cells at passage 2. The fAd-MSCs were characterized by flow cytometry, and their multipotential differentiation capacity into adipogenic, osteogenic and chondrogenic lineages, according to the criteria of the International Society of Cellular Therapy (ISCT), were evaluated using protocols previously published by our group [Further, fAd-MSCs of each donor were isolated and characterized according to a protocol previously optimized within our group ,29. Brieur group .2 in three T-175 flasks in DMEM supplemented with 10% fetal bovine serum (FBS), 2.5 mM L-glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin . When 80% confluency was reached, the cells were washed with PBS and DMEM with 10% exosome-free FBS was added to the flasks. The exosome-free FBS was previously obtained by ultracentrifugation at 100,000× g for 60 min at 4 °C. The culture medium containing the fAd-MSC-derived exosomes was collected after 72 h under standard culture conditions. The cells remaining in the flasks were quantified and their overall viability determined by trypan blue staining and loading in a Neubauer chamber.The exosomes from each individual donor were processed separately. The isolation of the exosomes was conducted by ultracentrifugation and their characterization afterwards was conducted following a protocol previously used by our group ,15. Brieg for 30 min at 4 °C to eliminate the cell debris and microvesicles. Afterwards, it was ultracentrifuged at 100,000× g for 90 min at 4 °C using a fixed-angle 70Ti rotor in an Optima LE-80K ultracentrifuge . The sedimented exosomes were collected by resuspending them in PBS before repeating the ultracentrifugation step. Finally, the exosomes were resuspended in 100 µL of PBS, quantified using a BCA kit  and stored at −80 °C until needed. For the characterization of the exosomes, equal amounts coming from each donor were combined.The culture medium was centrifuged at 13,000× Aliquots of the isolated exosomes were adsorbed onto formvar/carbon-coated 200 mesh grids, negatively stained with 2% aqueous uranyl acetate and observed using a JEM-1400 transmission electron microscope , operated at 80 kV following directions from the Transmission Electron Microscopy Unit from the Central Research Support Service (SCAI) of the University of Malaga.Size, distribution, homogeneity, and zeta potential of the exosome fractions (20 µL) that were previously diluted in 1 mL of distilled water were analyzed using a Malvern Zetasizer 2000 . Each sample was analyzed in triplicate to obtain the size distribution of the exosome population.The presence of exosome-specific biomarkers in the isolated fractions was evaluated by Western blot ,32,33,34The exosomal proteins were purified by a trichloroacetic acid protein precipitation-based procedure  and the resulting protein pellets were recovered and quantified by a Bradford assay. Afterwards, gel-assisted proteolysis was carried out, entrapping the protein solution in a polyacrylamide gel matrix. Briefly, gel plugs containing the proteins were cut into small pieces and incubated with DTT to break disulfide bonds. Cysteine residues were carbamidomethylated with iodoacetamide and the proteins were digested with trypsin . The resulting peptides were then extracted from the gel, dried in a SpeedVacTM vacuum concentrator, and re-dissolved in 0.1% formic acid aqueous solution. Finally, the peptides were purified using a C18 ZipTip  according to manufacturer’s instructions and brought to an identical concentration before being transferred to the injection vial.Samples were injected into an Easy nLC 1200 UHPLC system coupled to a hybrid linear trap quadrupole Orbitrap Q-Exactive HF-X mass spectrometer . The samples were automatically loaded onto a trap column  at a flow rate of 20 uL/min and eluted through a 50 cm analytical column . The binary gradient mobile phase consisted of 0.1% formic acid in water (solvent A) and 0.1% formic acid in 80% acetonitrile (solvent B). Peptides were eluted from the analytical column with a 120 min gradient ranging from 2% to 20% solvent B, followed by a 30 min gradient from 20% to 35% solvent B and finally, to 95% solvent B for 15 min before re-equilibration at a constant flow rate of 300 nL/min.m/z 300–1750 at a resolution of 120,000. The 20 most intense precursor ions were isolated within a 1.2 m/z window by a data-dependent acquisition method and fragmented to obtain the corresponding MS/MS spectra. The fragment ions were generated in a higher-energy collisional dissociation (HCD) cell and detected in an Orbitrap mass analyzer at a resolution of 30,000.Data acquisition was performed in the electrospray ionization positive mode. MS1 scans were acquired from ® HT as search engine and Felis catus NCBI version 2017.10.30 (43.896 sequences) as database. Peptide spectral matches and consecutive protein assignments were validated using the Percolator® algorithm [Acquired raw data were analyzed using the Proteome DiscovererTM 2.3 platform  with SEQUESTlgorithm  by imposLabel-free quantitation was implemented using the Minora feature of Proteome DiscovererTM 2.3. Protein abundances were based on precursor intensities. Normalization was performed based on total peptide amount and abundance ratio p-values were calculated by ANOVA based on individual protein abundances.http://www.geneontology.org/- accessed on 12 February 2021) using the network-based annotation service of the Proteome DiscovererTM 2.3 application.Biological process, molecular function and cellular component enrichment analysis of the identified proteins was carried out via the international standardized gene function classification system of gene ontology , CD34−, CD45−, and MHC-II- (hematopoietic markers) cell populations (data not shown). These fAd-MSCs differentiated to the three mesodermal lineages after appropriate induction. Adipogenic differentiation was evidenced by the presence of Oil Red O-positive fat drops; osteogenic differentiation was confirmed by alizarin red-positive calcium deposits; and chondrogenic differentiation was confirmed by the formation of micromasses exhibiting metachromasia when stained with toluidine blue .6 fAd-MSCs were isolated from the FCGS-positive cats and 7.12 ± 6.3 µg exosomes/106 fAd-MSCs from the healthy donors. The exosome fractions were analysed by TEM and dynamic light scattering (DLS). The exosomes that were isolated from the FCGS cats had a clearly lower average size and z-potential compared to the exosomes that were obtained from the healthy cats , hepatocellular function and development (oncoprotein-induced transcript 3), immune response (complement C1s subcomponent), regulation of biological processes , gene expression (threonine-tRNA ligase), vesicular transport , and protein folding (heat shock 70 kDa).The proteins that were, on the contrary, downregulated in the exosomes from the FCGS cats, were mainly involved in cell adhesion , mitochondrial metabolism (NADH-cytochrome b5 reductase), cell surface receptor (CD63), membrane and cytoskeletal dynamics (protein S100-A11), metabolic processes , regulation of biological processes , proliferation (guanine nucleotide-binding protein G), and RNA binding (poly(rC)-binding protein 2).The use of MSCs is an interesting therapeutic alternative for a large number of immune-mediated pathologies, due to the ability of these cells to modulate the immune system ,37,38. IIt has been estimated that around 50% of the fAd-MSCs might be infected with feline foamy virus , which sThe fAd-MSCs from the cats with FCGS produced notably more exosomes than the cells from the healthy donors , as has been previously reported for other pathologies . FurtherThe proteomic analysis of the exosome cargo, by high-resolution mass spectrometry, is a very useful tool for comparing the protein profiles in exosomes from different origins, and can lead to conclusions on their role in normal biological and physiopathological processes . A totalOn the other hand, 17 proteins were significantly more represented in the exosomes from the FCGS group, mainly related to metabolic processes, immune responses, regulation of biological processes, and gene expression. Thirteen other proteins were downregulated and these were mainly involved in metabolic functions.These differences confirmed that there are evident changes in the secretory profile of the fAd-MSCs, and the proteomic cargo of their exosomes, from cats with severe FCGS. The amount and variety of expressed proteins show that certain biological functions, related to tissue regeneration, metabolism, and immune responses, may be affected. These processes may be involved in the pathogenesis of FCGS and might help to understand what role the exosomes play in certain immune-mediated pathologies . FurtherAlthough complete teeth extraction usually leads to a high percentage of improvement in cats suffering from severe FCGS, around 30% of the patients are refractory to the existing treatments ,28. TherRecently, the use of both autologous and allogenic MSCs has been proven to be useful for the treatment of this pathology ,38. HencCurrently, the use of exosomes, as therapeutic agents in different disease models, has been shown to be equally as safe and efficient as the direct use of MSCs ,59. FurtThe number of animals studied, their age differences, and the limited amount of information available in the cat databases, are the main limitations of our study. Furthermore, since the exosomes from each donor could not be analyzed separately, possible inter-individual heterogeneity might have been lost. However, our results might set a base for the future identification of robust groups of proteins present in the fAd-MSC-derived exosomes from healthy and diseased animals. This would allow us to perform more studies on the use of exosomes and their cargo, for either diagnostic or therapeutic purposes.Finally, since the domestic cat suffers with FCGS naturally and in environmental conditions that are similar to those in which humans live, it might be a suitable preclinical model for transferring these results to some human pathologies, such as oral lichen planus, stomatitis, pemphigus, or pemphigoid .In this study, the alteration of the proteomic profile of fAd-MSC-derived exosomes, from severe, calicivirus-positive FCGS patients, is presented for the first time. Using bioinformatic tools, we have demonstrated that the altered proteomic profile in the exosomes from diseased patients affects the proteins related to several biological functions, such as tissue remodeling, metabolism, and immune responses. The exosomes do not only experience changes in their cargo, but are also produced in larger quantities. Possibly, the alterations in the exosomes in these patients will change their ability to participate in cell communication between the Ad-MSCs and their niche, which is affected by this pathology.


Background: The global epidemic of pediatric obesity is well-known, but data on co-existence of obesogenic behaviors are limited. We aim to report the prevalence of and trends in the co-existence of obesogenic behaviors in adolescents from 15 countries.Methods: This study was based on the Global School-based Student Health Survey 2003–2017 and included 121,963 adolescents aged 12–15 years from 15 countries where at least 2 cross-sectional surveys were conducted. We used sampling weights and calculated the country-level prevalence of and trends in the co-existence of obesogenic behaviors  during survey years. Pooled prevalence and trend estimates were calculated with random-effects models.Results: Pooled prevalence of exposure ≥ 1, ≥2, and ≥3 obesogenic behaviors was 88.2, 44.9, and 9.8% in the first survey and 88.4, 46.4, and 10.2% in the last survey, respectively. Plateauing, increasing, and decreasing trends in the co-existence of obesogenic behaviors were observed in different countries. Specifically, we identified a plateauing pooled trend in the exposure ≥ 1, ≥2, and ≥3 obesogenic behaviors .Conclusion: Trends in the prevalence of the co-existence of obesogenic behaviors varied significantly across different countries, but the prevalence remained high in most countries. These findings suggest the need for behavioral interventions to mitigate obesogenic behaviors in adolescents for overweight and obesity prevention. Obesity in children and adolescents is a major risk factor for physical and psychosocial disorders throughout the lifespan –5. The pThe onset and development of obesity is a complicated process affected by a variety of biological and behavioral factors , 8. CompSingle obesogenic behavior has been clarified well in adolescents , 10, 13.The GSHS is a series of multi-country, repeated cross-sectional, and school-based surveys , 10. It https://www.cdc.gov/gshs and https://www.who.int/ncds/surveillance/gshs/factsheets/en/. From the available GSHS dataset, 34 countries that underwent repeated cross-sectional surveys were included in the present study. We excluded 1 country that lacked a nationally representative sample; 1 country with a different coding rule compared to other countries; 2 countries where the last surveys were not conducted after 2010; 11 countries that lacked data on fruit and vegetable intake, anxiety-induced insomnia, physical activity, sedentary behavior or weighted information; and 4 countries that lacked a sufficient sample size  in any of at least 2 survey waves. Consequently, 177,201 school-attending adolescents from 15 countries were included in this study. We then excluded 46,029 adolescents whose age was missing or were >15 or <12 years old, and 9,209 adolescents for whom data on sex, food insecurity, fruit and vegetable intake, anxiety-induced insomnia, physical activity, sedentary behavior or weighted information were lacking. Eventually, 121,963 adolescents aged 12–15 years were included in the final analysis.Publicly available data from GSHS can be found at All students completed the survey questionnaire about obesogenic behaviors . Low vegThe GSHS asked participants for their obesogenic behaviors ; a value of “1” or “0” was assigned to participants who had or did not have each type of obesogenic behavior, respectively. Consequently, the number of obesogenic behaviors for each participant ranged from 0 to 4, with each type of obesogenic behavior worth 1 point.Data on sex, age, and food insecurity were also collected. Food insecurity was evaluated based on the question “How often did you go hungry because there was not enough food in your home during the past 30 days?” The response options were “never,” “rarely/sometimes,” and “most of the time/always , 10.”Sampling weights were used to account for the complex sampling design . We calcP-values of <0.05 as statistically significant.We used SAS version 9.4  and STATA version 11.0  to perform the statistical analyses, and considered This study included 121,963 adolescents aged 12–15 years from 15 countries. Ps > 0.05 for 10 countries; Ps < 0.05 for 3 and 2 countries). Notably, the prevalence of the co-existence of ≥2 obesogenic behaviors in all of the 15 countries in the latest survey year remained high (>30%), regardless of the different country-level trends. Furthermore, plateauing, increasing, and decreasing trends were identified in the prevalence of ≥3 obesogenic behaviors in 8, 5, and 2 countries, respectively . In 4 of the 8 countries with a plateauing trend  and 3 of the 5 countries with an increasing trend , the prevalence of ≥3 obesogenic behaviors in the latest survey year was >10%. This study indicated that the prevalence of exposure ≥ 2 and ≥3 obesogenic behaviors varied widely across the 15 countries and continued to be at high levels in most countries. We also founded the plateauing, increasing, and decreasing trends in the prevalence of exposure ≥ 2 and ≥3 obesogenic behaviors within different countries.The effects of obesogenic behaviors on adiposity are well-known. For example, low consumption of fruits and vegetables is known to promote pediatric obesity due to the displacement of fiber- and micronutrient-rich fruits and vegetables with energy-dense foods . A shortPrevious cross-sectional studies based on GSHS dataset and publications from other data sources have reported a high prevalence of single obesogenic behaviors , which is consistent with our results , 19–22. Our finding of the wide variation in the co-occurrence of ≥2 and with the ≥3 obesogenic behaviors and the corresponding trends across countries might be attributable to country-level variations in environmental, economic, social, policy-related, and other factors , 25–29. Multilevel factors, such as individual, family, community, and societal factors, contribute to the onset and development of childhood obesity . NotablyThe main strengths of our study are the nationally representative samples, rigorous and standardized data collection, the practical nature of this research and its global scale. However, some limitations need to be acknowledged. First, this study did not include countries from Europe or low-income countries, included only 1 country from the African region, and excluded 11 countries without complete data and 4 countries without the sufficient sample size  during any of at least 2 survey waves, which limits the generalizability of our findings. Second, the survey years, sample sizes, and survey numbers differed between the participating countries, which could have affected our results. Third, our study included only school-attending adolescents aged 12–15 years. A future study on trends in obesogenic behaviors should focus on younger children or adolescents who do not attend school. Fourth, there is no data about environmental factors that affect obesogenic behavior in the present study. Finally, the data collected were self-reported, which may have introduced some bias.In conclusion, the results of this study indicate that the prevalence of the co-existence of obesogenic behaviors remained high in most of the included countries during 2003–2017, even though the corresponding trends varied widely across the countries. This finding suggests that most countries need to implement interventions to mitigate obesogenic behaviors in adolescents and prevent overweight and obesity.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the local government administration and an institutional review board or ethics committee. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.HF conceptualized and designed the study, carried out the initial analyses, drafted the initial manuscript, and reviewed and revised the manuscript. XZ critically reviewed and revised the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The advancements in nanoparticles (NPs) may be lighting the sustainable and eco-friendly path to accelerate the removal of toxic compounds from contaminated soils. Many efforts have been made to increase the efficiency of phytoremediation, such as the inclusion of chemical additives, the application of rhizobacteria, genetic engineering, etc. In this context, the integration of nanotechnology with bioremediation has introduced new dimensions for revamping the remediation methods. Hence, advanced remediation approaches combine nanotechnological and biological remediation methods in which the nanoscale process regulation supports the adsorption and deterioration of pollutants. Nanoparticles absorb/adsorb a large variety of contaminants and also catalyze reactions by lowering the energy required to break them down, owing to their unique surface properties. As a result, this remediation process reduces the accumulation of pollutants while limiting their spread from one medium to another. Therefore, this review article deals with all possibilities for the application of NPs for the remediation of contaminated soils and associated environmental concerns. Rapidly increasing anthropogenic/technogenic activities are adding potentially toxic metals, agrochemicals, and an excess of nutrients to the soil . Soil isThe advancements in nanotechnology open a window globally to remediate or restore polluted soil in an effective way ,7. It haWith the help of NPs, hyperaccumulators and indigenous soil microbes could enhance biodegradation processes, thereby increasing the potential extent of remediation. This could be called nano-phytoremediation and microbial-mediated nano-remediation. The use of NPs with bioremediation approaches may result in a lot of benefits as these particles are small 1–100 nm) with a larger surface area and reactivity 0 nm with,12,13. TThe utilization of NPs based on a metal–organic framework (MOF) has received a lot of important attention lately, but it is primarily employed for drinking water and wastewater treatment ,17,18. TThe present review unbinds the possibilities for the elimination of contaminants, with particular emphasis on microbe-mediated remediation, hyperaccumulator plants, and NPs, as well as the approaches to restore HM-contaminated soils, the benefits, and the potential risks associated with nano-bioremediation technologies. The gaps and future perspectives are comprehensively elaborated.Soil rich in vital nutrients and micronutrients is believed to best support the optimum health of the plant and its growth . Human-mNanobioremediation is a cost-effective technique of utilizing plants and microbes for the breakdown of pollutant compounds, ultimately improving soil quality and reducing pollution. By breaking down contaminants in the soil, the process may be able to eradicate, retain, or reduce the amount of pollutants present ,25. The Nanobioremediation implies both nanotechnology and bioremediation together, where the process is executed at the nanoscale. The target pollutants are adsorbed, degraded, or modified owing to the unique physicochemical properties of the NPs, which also act as catalysts and help to reduce the activation energy required for breaking down the compounds . The nanBiosorption and bioaccumulation using plants and microbes are traditional methods to remove HMs from polluted soils. However, recent pieces of evidence have reported the use of NPs in HM remediation with remarkable outcomes . NanoparThe synergy of NPs and bacterial degradation has also gained attention; however, the availability of a handful of published papers would not permit the making of a categorical review. Many authors have given their efforts to nanobioremediation, and yet it is still too insignificant for making any conclusive decision .Integration of NPs with microbes for bioremediation is a two-phasic process that involves overlapping abiotic and biotic processes  36]. In. In36]. The existence of HMs in the environment is largely due to increased anthropogenic activities. However, disturbed biogeochemical cycles are also responsible for their release into the environment as pollutants. Elements like As, Cd, Cr, Hg, and Pb have no biological functions to perform in the biological system. Heavy metals comprise major inorganic pollutants as they exhibit substantial toxic impacts on biota even at the lowest concentrations ,41. The Heavy metals primarily affect the plants and lower soil organisms by inducing the generation of reactive oxygen species (ROS), which further results in the damage of macromolecules such as proteins and nucleic acids . The exiMorganella psychrotolerans [One of the key methods for the elimination of HMs includes site stabilization that immobilizes them at a specific site to decreases mobility and availability in the soil, and stops them from leaching across the sites . The usetolerans ,58.Halomonas sp. This approach has significantly removed nearly 100% of Pb after 24 h, and Cd after 48 h, as compared to removal by bacteria or only NPs [3O4 NPs treated with S. aureus, with a surface encapsulated with phthalic acid (as a n-Fe3O4-Phth-S complex), was used for the removal of Cu, Ni, and Pb, and the adsorptive removal of 795, 1355, and 985 µmol g−1 for Cu, Pb, and Ni was achieved, respectively. In terms of percentage, the recovery rates of 83.0–89.5% for Cu2+, 99.4–100% for Pb2+, and 92.6–7.5% for Ni2+ were observed. The comparative study with dried S. aureus and n-Fe3O4-Phth-S for HM removal inferred that the n-Fe3O4-Phth-S core of the NPs, as well as the functional groups present on the microbial surface, played a key role in the removal of HMs [Nanoparticles of FeO coated with polyvinylpyrrolidone (PVP) have been successfully used for improving the bioremediation of the soil contaminated with Pb and Cd by a Gram-negative bacteria, only NPs . A biosol of HMs . Thus, tB. cereus (PMBL-3) and L. macroides (PMBL-7) evidently confirmed that ZnO NPs at 5 mg L−1 synergistically removes the Cr by 60%, the Cu by 70%, and the Pb by 85%, as compared to B. cereus (80 and 60%) and L. macroides (55 and 50%) at neutral pH, respectively [B.cereus has also been reported to reduce the Cr6+ through an enzyme-mediated process. The reduced Cr3+ was observed to have a binding affinity to cells using coordination bonds with the functional group present on the surface of the bacterial cell wall. The formation of Cr2O3 NPs was found on the cell surface as a by-product [A recent study on the removal of Cu, Cd, Cr, and Pb using HM-resistant bacteria such as ectively . At neut-product .L. casei and L. fermentum) to absorb Cd from water in association with Se5+ and Se NPs was also investigated. The higher absorption of Cd by L. casei with Se4+ ions (65%), compared to Se NPs (55.90%), was discovered in this study, and it was correlated to the higher solubility of Se5+ compared to Se NPs. When comparing L. fermentum and L. casei, the efficiency of Cd absorption was significantly higher in L. fermentum (50.87%) than L. casei (43.78%). The percentage of Cd adsorption by L. casei when used in conjunction with Se NPs shows no significant change. However, with increased Se NPs ratio percentages, Cd absorption was slightly increased from 5.49 to 16.54 in the presence of L. casei with Se NPs, compared to L. casei [The use of probiotic bacteria . The range of Pd NPs was observed as 10 nm by transmission electron microscopy; however, the size of the Pd NPs was dependent on environmental conditions such as temperature, pH, and biomass. The obtained Pd NPs have great use as a bionanocatalyst that shows good catalytic efficiency (6.3 mg Pd NPs completely reduced 5.0 µmol Cr6+ in 12 h) and the application is potentially useful to treat industrial effluents [A threefold approach is now gaining popularity as the HM pollutants can be used by selective microbes to synthesize biogenic NPs (resource recovery), thereby removing them from the environment (remediation) and yielding value for the waste (effective waste utilization). A study using ffluents .0 NPs) using pollutant tellurite Te4+ oxyanions present in the wastewater. It has been reported that 2-Hydroxy-1,4-naphthoquinone promotes the reduction of Te4+ and the quantity of Te0NPs synthesis [Rhodobacter capsulatus, where malate is the electron-donating substrate [4+ reduction by anaerobic methanogenic granular sludge [A similar study produced Te NPs from anaerobic sludge based upon supplementation with riboflavin . It formynthesis . The proubstrate , and ribr sludge .Sphingomonas) was earlier reported to degrade the pesticide hexachlorocyclohexane (HCH) [Sphingomonas sp. strain NM05 or CMC-Pd/nFe0 alone [The pollution posed by POPs has been shown to have a negative influence on both the environment and human health, as certain POPs have been found to bioaccumulate in adipose tissue and to have the potential to act as carcinogens. Therefore, their remediation is a major challenge and is obligatory. A Gram-negative bacterial strain (NM05 of ne (HCH)  upon tree0 alone . The dege0 alone .3) NPs and biochar from water caltrop (Trapa natans) shells studied on marine sediment reported enhanced degradation of PAHs. The study used lignocellulosic fiber-reinforced biodegradable composites (LFBC) at 0.75 g L−1 and pH 6.0 to activate the peroxymonosulfate (3 × 10−4 M) that helped in the oxidation of oxidizing PAHs in the sediments.The perovskite  from column effluent within 50 days [Up to 90% of total degradation was achieved; however, individually 2-ring PAHs 52%, 3-ring PAHs 61%, 4-ring PAHs 66%, 5-ring PAHs 56%, and 6-ring PAHs 29% were observed . The proobserved . In a co 50 days .In the case of soil, improving the microbial community by application of NPs is another way to reduce/remove the toxic pollutant loads from it. Si NPs have been reported to improve microbial colonization and biomass, including the rhizospheric microbes that are helpful for improving soil health ,73. HoweBoth from an ecological standpoint and one of restoring degraded areas, the remediation of HM-contaminated soils is a pressing issue that needs to be addressed immediately . This se2 and O2, etc. [The kinds of soil found at a contaminated site, as well as the percentage of metal contamination present, influence the rate at which hyperaccumulating plants can be utilized to remediate the site . The bioO2, etc. . The rhiO2, etc. .Pedioplanis burchelli, Amaranthus spinosus, and Alternanthera pungens, have survived at an optimal level of HMs by rhizofiltration and demonstrated avoidance mechanisms for HM uptake in their environment [A few plant species, such as ironment . Beyond ironment . Howeverironment ,79. The ironment .Parthenium hysterophorus, Sacrum munja, and Ipomoea carnea, and the authors observed more than one BCF and TF for Cr, Ni, Cd, and Pb, which revealed a phytostabilization mechanism, while more than one BCF and less than one TF for Zn and Mn showed a phytoextraction process for HMs [The hyperaccumulator plant that has a bio-concentration factor (BCF) of HMs of more than one owes it to the mechanism of phytostabilization and phytoextraction . A BCF a for HMs . SchematRhizodegradation is the process through which pollutants are deposited in the rhizospheric area of soil by microbial activity, where bacteria metabolize these contaminants for energy and nutrition. In this mechanism, microbes are capable of breaking down hazardous pollutants into nontoxic and harmless products . Plant rA wide range of treatment strategies, including physicochemical and biological methods, have been used to decontaminate sites polluted with HMs. The mechanisms of these methods are based on redox reactions, adsorption, ion-exchange, bioremediation, and phytoremediation . All theRecently, adsorbent materials such as activated carbon, biochar, and NPs became commonly available; these adsorbent materials exhibit rapid adsorption capacity, cover large surface areas, provide more interplay sites for HMs, and have low price value . Hence, 2 NPs, and Ag NPs have all been extensively investigated for their potential for soil remediation [2+ at 10.0 pH [2+ are removed at 3.0 pH [6+ at 6.0 pH, while at high pH (>10.0) the FeS NPs elimination rate was decreased [2+ by Fe3O4 NPs rises from 298 to 328K [2+ at 30 °C [2+ removal efficacy by FeS NPs (stabilized by sodium carboxymethyl cellulose) achieves its highest elimination percent at 30 min of contact time [2+ and Fe3+, which develop the excellent potential of nZVI for the elimination of HMs. Removal of Zn2+ (109.7 mg g−1), Cu2+ (161.9 mg g−1), and Pb2+ (195 mg g−1) was achieved by utilization of nZVI after 6 h [For the remediation of soil contaminated with HMs, carbon or non-carbon NPs have primarily been employed; CNTs (carbon nanotubes), nZVI , TiOediation . The HM- 10.0 pH  and a hit 3.0 pH ; FeS NPsecreased . The equ to 328K , while cat 30 °C . Hg2+ react time . The nZVnd Pb2+ 1 mg g−1 w−1) was utilized in the pot experiment, and it was observed that the immobilization efficiency of Cr6+ from the soil was increased by 100% after 15 days [−1) by 46.66%, 48.88%, and 47.01%, respectively, reported [Salix alba L.) with an increased bio-concentration factor for Cd, while a high concentration of nZVI showed an adverse effect on plant growth and BCF for Cu and Pb [Brassica napus) and Chinese cabbage (B. rapa subsp. pekinensis) [Several HMs  are effectively removed by the application of nZVI at the highest rate from contaminated sites . The nZV 15 days . The elireported . The lowu and Pb . The eliu and Pb  using nZ2 NPs have high reactivity and photo-catalytic activity, which enables contaminates to get adsorbed on its surface area; this trait of TiO2 NPs serves to minimize the toxic behavior of HMs, as well as maintain their mobility. It absorbed 88.01% of Cu2+ and 70.67% of Cd2+ effectively at 7.0 pH [Suaeda salsa and decreased the uptake of Cd and Ni compared to the control [Zea mays) plants and promote plant growth by optimizing the gibberellin, abscisic content in Z. mays leaves and PGPR interaction [B. cereus (LPR2), Ag NPs significantly induce the growth of Z. mays [It is observed that TiOt 7.0 pH . Other N control . Ag NPs eraction . In comb Z. mays . Hence, Metalloids are elements with a plurality of attributes, i.e., they have the physicochemical features of both metals and nonmetals. They are naturally found, but the introduction of industrialization has resulted in concentrations that exceed the allowable limit . AntimonIn previous years, several remediation methods have been introduced for the reclamation of these metalloids from soil. However, successful remediation of sites contaminated by metalloids is hindered by the fact that these pollutants do not decompose on their own and that it is not always possible to extract all of the contaminated soil . Therefo3O4 particles for immobilizing As in contaminated soils. Among these, Fe3O4 NPs were reported to be more effective than other NPs for immobilizing As. Besides iron oxides, other NPs are also described as remediating soils polluted with different metals and metalloids [6+ to non-toxic Cr3+ in aqueous [6+ reduction efficiency was recorded up to 90%. Furthermore, in some cases, NPs were also employed to immobilize the trace elements in contaminated soil. For example, nanostructured TiO2, MgO, and ZnO were found to be efficient adsorbents for the elimination of Cr ions from leather factory waste treated soil [Among NPs, iron oxides NPs are considered to remediate contaminants like HMs from soils owing to their capacity to absorb these pollutants . In thistalloids . Apart f aqueous . In the ted soil . Hence, ted soil ,115. AltDespite the numerous roles of and advancements in nanotechnology, there is still concern about its presence in environmental spheres, its fate, and the consequent toxicological impacts. Therefore, the main focus of this section is to distinctly provide an overview on the fate and the emerging environmental challenges of the deliberate emission of NPs, owing to their enhanced applications, especially in remediation processes. Recently, some reports documented that the increasing applications of NPs in agriculture have imposed serious implications. For example, the deliberate application of NPs may result in their accumulation or in an increase in the concentration of their constituents in the soil, thus affecting the soil’s properties ,20. The + from the soil to yield Cu ions and Cu(OH)+ ionic complex. Furthermore, this process is stated to be more enhanced in acidic soil [A study conducted by Cullen reported that modification of nanoscale zero-valent iron (nZVI) in the soil can cause a significant rise in the pH of a soil solution . A simildic soil . Also, Ndic soil . The extdic soil . In addidic soil ,121. Furdic soil .In soil, there are three main procedures, viz., surface complexation, hydrophobic partitioning, and ion exchange for the sorption of NPs, which are also involved in the colloid-facilitated transport of pollutants . MobilizDanio rero (zebrafish), Daphnia pulex (daphnia), and Pseudokirchneriella subcapitata , and they caused acute toxicity and filter-feeding, according to the findings of the study. It was noted that the NPs were more hazardous to daphnia and microalgae than to zebrafish, which is understandable because daphnids are particle filter feeders and, hence, are more susceptible to being exposed to NPs [2 NPs and TiO2 NPs induced immunomodulatory and immunosuppressive effects in biological systems [Once NPs enter into water bodies, they act as pollutants and are ingested by lower aquatic biota. Metal NPs were tested in three different animal models representing different trophic levels, including the d to NPs . Thus, ad to NPs ,137. DNAd to NPs ,139,140. systems ,142. ThuIn this modern period, there is a rapid release of pollutants into the environment, posing a substantial hazard to both human and the environmental health. As a result, having effective techniques for removing them from various environmental media will be critical in preventing their negative impacts. Because many conventional procedures are incapable of efficiently removing different groups of contaminants, novel approaches are therefore required to eliminate pollutants to the maximum extent possible. Meanwhile, a very likely method for developing remedies for polluted media is nanotechnology, according to the observations from a profusion of studies in this respect. A favorable result is the filling of the data gap on real-world remediation, which is aided by closer collaboration between the research and the industry. Thus, it will be useful for driving future soil cleanup efforts, which are sometimes necessary and cannot wait for experimental results to be obtained, utilizing methods to be attained in the meanwhile. Besides, the majority of the currently available research on nanobioremediation is limited to laboratory experiments and computational modelling. Therefore, to alleviate soils contaminated with a varied range of contaminants in the field, it is necessary to utilize multidisciplinary approaches. However, the environmental fate of NPs should be carefully taken into consideration before widening the application of nanobioremediation approaches.