Since the rise and rapid development of nanoscale science and technology in the late 1980s, nanomaterials have been widely used in many areas including medicine, electronic products, crafts, textiles, and cosmetics, which have provided a lot of convenience to people’s life. However, while nanomaterials have been fully utilized, their negative effects, also known as nano pollution, have become increasingly apparent. The adverse effects of nanomaterials on the environment and organisms are mainly based on the unique size and physicochemical properties of nanoparticles (NPs). NPs, as the basic unit of nanomaterials, generally refer to the ultrafine particles whose spatial scale are defined in the range of 1–100 nm. In this review, we mainly introduce the basic status of the types and applications of NPs, airborne NP pollution, and the relationship between airborne NP pollution and human diseases. There are many sources of airborne NP pollutants, including engineered nanoparticles (ENPs) and non-engineered nanoparticles (NENPs). The NENPs can be further divided into those generated from natural activities and those produced by human activities. A growing number of studies have found that exposure to airborne NP pollutants can cause a variety of illnesses, such as respiratory diseases, cardiovascular diseases, and neurological disorders. To deal with the ever increasing numbers and types of NPs being unleashed to the air, we believe that extensive research is needed to provide a comprehensive understanding of NP pollution hazards and their impact mechanisms. Only in this way can we find the best solution and truly protect the safety and quality of life of human beings. The development of nanotechnology can be traced back to the 4th century AD, and the most iconic example is the Lycurgus cup made of dichroic glass by the Romans. The colloidal gold and silver in the glass give the Lycurgus cup an opaque green color in reflected light, but a translucent red color in transmitted light ,2. The UNanoparticles (NPs), as the basic unit of nanotechnology products, generally refer to the ultrafine particles with a size less than 100 nm in at least one spatial dimension ,4. So whWhile people are intoxicated with the many novel functions of nanomaterials and the wonderful application prospects they will bring to our life, their potential safety problems are also increasingly emerging. Due to the extremely small size of NPs, there is a high possibility of inhalation during the development, production, use, and recycling of nano products. It is also a reminder that we need to reassess and understand how they are absorbed and what biological effects they can cause. In addition to the previously mentioned NPs produced by artificially controlled engineering processes, namely as engineered NPs (ENPs), inhalable NP pollutants also include NPs produced by gradual degradation of physical, chemical and biological processes in the environment, which are generally referred to as non-engineered NPs (NENPs). A number of studies have found that airborne NPs are able to penetrate deep into the alveoli through the respiratory tract following inspiratory action, and then distribute along the circulatory system to various tissues and organs, including the liver, spleen, kidneys, heart, and brain, where they may be deposited ,9,10,11.This review will elaborate on the basic status of the types and applications of NPs, airborne NP pollution, and the relationship between airborne NP pollution and human diseases.As mentioned earlier, the common feature of all ENPs is that at least one spatial scale is in the 1–100 nm range ,4. AccorThe main component of the inorganic NPs family is inorganic matter. Because of their abundant constituent elements and various kinds, they have diversified excellent performance. Here we will only focus on some of the classical ENPs, such as carbon-based NPs, metal-based NPs and oxide NPs.The carbon-based NP family is a family mainly composed of carbon elements, and their morphologies have been in constant change and development. According to the division of spatial dimension, carbon-based NPs generally include the following categories: zero-dimensional  and fullerenes), one-dimensional  and multi-walled carbon nanotubes (MWCNTs)), two-dimensional , and three-dimensional  . In thisC-dots, also known as carbon quantum dots or carbon nanodots, are typically zero-dimensional nanoscale carbon nanomaterials, whose morphology is a grape-like aggregate composed of highly fused spherical carbon particles, with a very small size (below 10 nm). Its history can be traced back to 2004, when Xu et al., from the University of South Carolina in the United States, first discovered C-dots that can emit bright fluorescence during the electrophoretic purification of products in the preparation of SWCNTs . C-dots 60) is one of the most widely known. It is a perfectly symmetrical caged icosahedron about 1 nm in size, made up of 60 carbon atoms with 12 pentagonal rings and 20 hexagonal rings [60 mainly focuses on its catalytic performance, superconductivity, biocompatibility and oxidation resistance. For example, Li and Xu used C60 as catalyst to catalyze the hydrogenation of nitrobenzene to aniline, in which the catalytic hydrogenation efficiency was as high as 100% [60 as a catalyst with excellent performance. In addition, the discovery of doping C60 superconductor is another major achievement in the field of superconductivity. Tanigaki et al. obtained superconductors with good performance and a critical temperature only second to that of oxide ceramic superconductor by doping C60 in metal potassium and rubidium [60 “free radical sponge” because of its role in scavenging reactive oxygen radicals, activating skin cells and preventing aging [60 is also used in cosmetics and skin care products and other related industries [60 in the disease treatments, such as anti-inflammatory, antiviral, or antibacterial agents [Fullerenes, discovered in 1985 by Harold Kroto of the University of Sussex in collaboration with James Health of Rice University in soot residue from the gasification of carbon in helium gas flows, are another zero-dimensional carbon-based NP material ,88. Throal rings . It is a as 100% . This oprubidium . Therefong aging . Therefodustries ,23. Othel agents ,25,26,27l agents ,31, as wl agents .3–108 [CNTs are hollow cylindrical nanomaterials formed by the graphene sheets composed of carbon six membered rings curling in one direction. Therefore, according to the number of graphene sheets, they can be roughly divided into two categories: SWCNTs formed by a single graphene sheet and MWCNTs formed by several concentrically nested graphene sheets. The nanoscale of these two types of CNTs is reflected in the radial dimension. The diameter of SWCNTs is extremely short, some only about 1 nm, while the diameter of MWCNTs generally varies from a few nanometers to tens of nanometers . Their l3–108 ,91. Sinc3–108 . For exa3–108 ,37,38, g3–108 ,37,39, a3–108 , and eve3–108 ,94 to tr3–108 ,42,43. I3–108 ,34. In a3–108 .2 hybrid carbon atoms covalently bonded in a hexagonal lattice. As early as 1895, Brodie discovered the presence of highly layered structures in thermally reduced graphite oxide [Graphene is actually an atomic layer of graphite, that is, a two-dimensional monolayer formed by SPte oxide . It was te oxide . Graphente oxide . It is lte oxide . Therefote oxide ,47,48,49te oxide ,45,46.Metal-based NPs are another important part of inorganic NPs family. In fact, in terms of composition, Metal-based NPs are exactly the same as ordinary metal materials; the key difference is that the particles or thickness of nano metal are on the nanoscale, so that nano-sized metal particles have nano properties that traditional metals do not! Common metal-based NPs include gold nanoparticles (AuNPs), silver nanoparticles (AgNPs), copper nanoparticles (CuNPs), and platinum nanoparticles (PtNPs). It is worth mentioning firstly that metal-based NPs and oxide NPs to be mentioned later are different from the carbon-based NPs. Each type of NPs comes in a variety of shapes and sizes ,100,101,165) and basic fibroblast growth factor (bFGF) [Compared to bulks of gold, AuNPs and their applications may be considered to be the product of modern science. However, as previously mentioned, AuNPs and AgNPs were used to color the Lycurgus cup, made by the Romans in the 4th century AD, making it a rare color-changing glass, that appears opaque green in reflected light while translucent red in transmitted light ,2. It war (bFGF) ,51 and pr (bFGF) ; or by er (bFGF) ,54,55,56r (bFGF) ,60,61. Tr (bFGF) .Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, Candida albicans, and other hundred kinds of bacteria [AgNPs as bacteriostatic agent is a major feature of their applications. In fact, silver itself has a certain antibacterial effect. However, the antibacterial activity of silver particles at nanometer size is much stronger than that of silver. A small amount of AgNPs can produce a powerful bactericidal effect. This bactericidal effect has a broad spectrum and can inhibit and kill including bacteria . Even bebacteria ,67,68. Abacteria . In addibacteria ; or by ubacteria .Compared with AuNPs and AgNPs, CuNPs and PtNPs seem to have better catalytic applications. The high surface area of NPs enables them to provide better dispersion of the active site and easy diffusion of the reactants. CuNPs and PtNPs not only have good catalytic performance in their own reactions, but also their catalytic performance can be significantly improved when they are combined to form bimetallic catalytic materials ,71. In a2) NPs, silica (SiO2) NPs, zinc oxide (ZnO) NPs, iron oxide (Fe2O3) NPs, and alumina (Al2O3) NPs. As mentioned earlier, oxide NPs, like metal-based NPs, have a variety of structural forms, including spherical, rod, cube, and so on [2 NPs, SiO2 NPs, and ZnO NPs are the three most produced nanomaterials worldwide. TiO2 NPs are widely used in solar cells, paints and coatings due to their photolysis properties. In addition, TiO2 and ZnO NPs not only absorb ultraviolet light, but also reflect and scatter ultraviolet light, so they are used in cosmetics such as sunscreen [2 NPs are widely used in cosmetics, food, automotive industry, coatings, and even biomedical fields [2O3 NPs makes them suitable for use in magnetic separation of biological products and cells, diagnosis and guidelines for site-specific administration [2O3 NPs are widely used in strengthening and toughening plastics, rubber, ceramics, refractories, and other products [In addition to carbon-based NPs and metal-based NPs, oxide NPs are also an important part of inorganic NPs family. There are many types of oxide NPs, including titanium oxide , particles with aerodynamic diameter less than 2.5 microns , and particles less than 100 nanometers . In fact, the health effects of the first two types of particulate pollutants are better understood. However, with the wide application of ENPs, they began to be accumulated in the environment, which gradually aroused people’s strong attention on their role in human health and environmental pollution. In this section, the main sources of airborne NPs are described in detail , about 7 million people worldwide die from air pollution each year . Particun detail .3 in a CNT production laboratory [3 in an injection chamber at a AgNPs manufacturing facility [3 for AgNPs inhalation recommended by the American Conference of Industrial Hygienists [At present, airborne NP pollutants can be divided into two categories according to their sources: ENPs and NENPs. ENPs are made by artificially controlled engineering processes. As a result, exposure to ENPs can occur in a range of activities from the manufacture of nano products , product installation and use, and product disposal and recycling. For example, Han et al. detected airborne dust containing CNTs at levels up to 400 μg/mboratory . Lee et facility , which ogienists . Exposur3 [3 compared to lower background values (<4000 particles/cm3). In addition, Guyon et al. sampled within the boundary layer of 69 samples from biomass combustion in the Amazonia and estimated NENPs diameters of 110 ± 15 nm with emission factors ranging from 2.3 × 1014 to 5.4 × 1015 particles emitted per kg dry matter burned [NENPs are particles produced by gradual degradation in the environment through physical, chemical, and biological processes. While there are many differences in the physicochemical composition of ENPs and NENPs, one common feature is their very small size , which gives these particles unique properties that may cause harmful effects on human health. Generally, NENPs can be also divided into two categories: naturally occurring NENPs and artificially produced NENPs. Due to natural phenomena such as dust storms, volcanic eruptions, and forest fires, NENPs are already abundant in nature, which could have a serious impact on air quality. As Taylor reported in 2002, naturally occurring aerosols account for 90% of the total, while human activity generates only about 10% of the total. Dust storms appear to be the biggest source of naturally occurring NENPs . Accordi3 . Before 3 . Concentr burned .5 particles/cm3, and that most NENPs are aggregates of small spheres 30–50 nm in diameter. In addition, Kim et al. and Graczyk et al. tested NENP concentrations in a rubber factory and welding plant respectively and found that the NENP concentrations in the environment were very high, posing a very high risk of occupational exposure [5 particles/cm3 in the rubber factory and 1.69 × 106 particles/cm3 in the welding plant, with average diameters of 26 nm and 45 nm, respectively. NENPs generated in transportation are mainly caused by incomplete combustion of diesel or gasoline. The diameter of NENPs produced by diesel engines is mainly in the 20–130 nm range [11 and 3.2 × 1012 particles/vehicle/km at speeds of 50 and 70 km/h [7 particles/cm3 before and 2.0 × 103 particles/cm3 after the filtration treatment. The average diameter of the pre-filtered NENPs was about 150 nm, while the average diameter of the filtered NENPs was smaller, around 90 nm. This shows that the fabric filter does have a strong purification effect on the waste gas, but the open burning of household garbage still exists and causes great harm to the environment, suggesting that we still need to deal with garbage more properly. As a combustion product of smoking, tobacco smoke is also one of the manufactured air pollutants containing NENPs. It has been reported that, under poor ventilation, smoking can significantly increase the concentration of airborne particles in rooms and buildings [2.5 concentrations from 93 smoking homes were 31 μg/m3 and 3 μg/m3 for the 17 non-smoking homes, suggesting that members of smoking homes were at higher risk of fine particle exposure [11 particles/min, and the median diameters of particles were 102–113 nm [9 particles/cm3, and the particle size was in the 120–165 nm range [Artificially produced NENPs are also mainly produced through high temperature and combustion, and their sources include industrial emissions, transportation emissions, waste incineration, human routine activities, and smoking. The concentration and size of NENPs emitted by industry are determined by the raw material, processing manners and emission treatment. Leoni et al. measured NENP concentrations in residential areas 1.5 km away from a large steel plant . They esexposure ,116. Thenm range , while tnm range . Wearing 70 km/h . In addi 70 km/h . They fouildings ,122. Bas2–113 nm . Similar2–113 nm . Fuoco enm range . In addinm range ,127,128.10) entering with air mainly stays in the nasal cavity and upper respiratory tract, while fine particulate matter (PM2.5) and nano particulate matter (PM0.1) with small particle size may penetrate deeper into the alveoli [0.1 may also penetrate different tissue compartments of the lung, eventually reaching capillaries and circulating cells. These particles then travel through circulation to other organs, including the liver, spleen, kidneys, heart, and brain, where deposition may occur. As a result, not only does the respiratory system suffer damage from airborne NP pollutants, but other tissues and organs of the human body are also at risk  and large, thick MWCNT  induced strong acute phase and inflammatory responses after a single intratracheal instillation in C57BL/6 mice, respectively [In fact, inflammatory response is a defense mechanism activated by organisms in the face of “exogenous” foreign bodies. As a non-specific response, it can resist any kind of attack, whether biological, chemical or physical. In general, inflammation is a beneficial adaptive response that helps to maintain the body’s integrity. However, when the foreign body cannot be removed, a transition to chronic inflammation may occur, resulting in different degrees of pathological changes. There is no doubt that NPs, once in the body, are also recognized by the organism as foreign bodies that must be eliminated by inflammatory response, thus causing inflammation. For example, Poulsen et al. found that both small, curled , and calcitonin gene-related peptide (CGRP) were significantly increased in bronchoalveolar lavage fluid in asthmatic mouse models exposed to TiO2 NPs [2 NPs. These results suggest that TiO2 NPs can promote the development of neuroinflammation, and the enhanced neuroinflammation may be involved in the pathogenesis of bronchial asthma, leading to the release of neurotransmitters, thus aggravating asthma.Lavigne et al. conducted a population-based cohort study in Toronto, Canada, to assess the association between exposure to NENPs during pregnancy and early postpartum and the incidence of childhood asthma . In modeTiO2 NPs . At the In addition, as previously mentioned, CNTs have extremely high aspect ratio properties similar to those of asbestos fibers. There are concerns that inhaling CNTs may also cause damage similar to asbestos fibers, such as pulmonary inflammation and fibrosis. When Shvedova et al. treated C57BL/6 mice with pharyngeal aspiration of SWCNTs (1–4 nm in diameter), they found some abnormal responses in the mice’s lungs, which combined robust but acute inflammation with early onset yet progressive fibrosis and granulomas .10/PM2.5 and cardiovascular diseases, it is natural to think that the airborne NP pollution also plays a role in the process of atherosclerosis, myocardial infarction, and other diseases.Unlike respiratory diseases, the association between environmental particulate exposure and cardiovascular diseases was not accepted until the mid-1990s, when it was observed that hospital admissions for cardiovascular diseases increased on days with higher particulate concentrations . Since t2) NPs (5 nm in diameter) would cause oxidative stress and inflammation in the lungs and cardiovascular system, and such stress would eventually exacerbate the progression of atherosclerosis in the apoproteinE-deficient (ApoE−/−) mice, a sensitive animal model [2 NPs exposed mice were significantly larger than those in the control group (1.8 times), and the transcript levels of several genes involved in atherosclerosis, CCL-2, vascular cell adhesion molecule 1 (VCAM-1), and cluster of differentiation 68 (CD68) were also increased.Miller et al. used AuNPs to evaluate particle translocations when exploring the relationship between NPs and cardiovascular diseases . In miceal model . The res60, leading to exacerbation of myocardial infarction [Later, studies found that exposure to NPs also had a certain aggravation of the symptoms of myocardial infarction. After treating myocardial infarction mice with ZnNPs, Song et al. observed that ZnNPs exposure increased the size of myocardial infarction cells and disordered cell arrangement . Thompsofarction .2 NPs [In addition to the above diseases, the nervous system can also be affected by exposure to airborne NP pollutants. The earliest evidence comes from a study by Kilburn, who found that workers exposed to diesel had deteriorated visual performance, impaired cognitive performance, and balance problems, suggesting they had chemical brain damage . Later, 2 NPs . A numbe2 NPs ,146. Ame2 NPs . Further2 NPs .As mentioned above, NP materials do have a huge impact on people’s lives in many ways. While these materials are being developed and applied in large numbers, NPs are also beginning to make their way into the environment. NPs come from a wide variety of sources, whether unintentional or intentionally created, and if they are not managed and controlled, the situation of airborne NP pollutants will only be worsening. People’s exposure to airborne NP pollutants increases as these pollutants accumulate more in the air. Airborne NP pollutants enter the human body through the respiratory system, and then distribute to various tissues and organs along with the circulatory system. Accumulated day after day, their toxic effects will cause certain damage to the human body, such as the diseases mentioned above. Although many recent studies have focused on the adverse effects of airborne NP pollution, serious knowledge gaps remain in definitively identifying the human health effects of exposure to these nanoscale pollutants and the mechanisms of action involved. At present, the solutions to NP pollution are mainly reflected in the following aspects: technological innovation, policy formulation, and enhancing civic awareness. Technological innovation mainly depends on scientists’ exploration and research. There are also some possible directions. For example, we hope to find safer and more environmentally friendly NPs to replace those that are harmful to humans and the environment, or introduce some special enzymes, bacteria, and fungi to treat nano-waste. Policy formulation is mainly aimed at the control of the production process of NPs. In addition, raising citizens’ awareness of NP pollution is also a meaningful initiative. In conclusion, until the issue of NP contamination is identified and addressed, it is necessary to strengthen safety management of NP sources in order to reduce exposure and protect human health.


Curtobacterium, belonging to the Microbacteriaceae family of the Actinomycetales order, includes economically significant pathogenic bacteria of soybeans and other agricultural crops. Thorough phylogenetic and full-genome analysis using the latest genomic data has demonstrated a complex and contradictory taxonomic picture within the group of organisms classified as the Curtobacterium species. Based on these data, it is possible to delineate about 50 new species and to reclassify a substantial part of the Curtobacterium strains. It is suggested that 53 strains, including most of the Curtobacterium flaccumfaciens pathovars, can compose a monophyletic group classified as C. flaccumfaciens. A genomic analysis using the most recent inventory of bacterial chromosomal and plasmid genomes deposited to GenBank confirmed the possible role of Microbacteriaceae plasmids in pathogenicity and demonstrated the existence of a group of related plasmids carrying virulence factors and possessing a gene distantly related to DNA polymerase found in bacteriophages and archaeal and eukaryotic viruses. A PCR diagnostic assay specific to the genus Curtobacterium was developed and tested. The presented results assist in the understanding of the evolutionary relations within the genus and can lay the foundation for further taxonomic updates.The genus of  Curtobacterium, in 1972 [Curtobacterium comprised eight validated species, recently accompanied by a number of proposed species and unclassified strain groups. Phytopathogenic strains causing the wilting and rotting of various legumes (Fabaceae) and ornamental plants have been established as a separate species, C. flaccumfaciens, and further subdivided into several pathovars, depending on the host plants and physiological properties [Curtobacterium spp. are not known to cause any disease of plants from which they were primarily isolated [Since the definition of the genus operties . Meanwhiisolated . Many stisolated , grapeviisolated , maize [isolated , tomatoeisolated , black pisolated , strawbeisolated , citrus isolated , poplarsisolated  and eucaisolated  and haveisolated  and mariCurtobacterium spp. in plant pathogenesis worldwide and the links between genomic features and virulence with respect to plants used for taxonomic distribution need elaboration and revision.Therefore, the role of Curtobacterium using data acquired from previous taxonomic studies. Polyphasic studies largely depend on the 16S rRNA gene [It is difficult to unravel the structure of the complex genus RNA gene . ClassifRNA gene .New taxonomic technologies often raise the question as to whether discrepancies associated with earlier phylogenetic structures were caused by conflicts between the phenotype, single genes or entire genomes or by data interpretation, such as those between the different algorithms of a taxonomic analysis . In phylCurtobacterium based on the principles of phylogenetic systematics applied for genome comparison. A comprehensive sampling of publicly available whole-genome sequences of strains representing the genus Curtobacterium was used to construct genome-scale phylogenetic trees and to address the following questions:(a)Curtobacterium?Is there any conflict between the phylogenies calculated from whole-genome sequences and the current classification of the genus (b)Curtobacterium need to be revised because they are evidently nonmonophyletic?Which species within the genus The present study was designed to provide an improved framework for the classification of the genus Curtobacterium plasmids have been shown to contain possible virulence genes, and the effect of mobile elements and gene exchange could reveal the ways of the emergence of pathogenic strains and pathovars [Curtobacterium plasmids were studied in the context of the genomics of pathogenicity.Another important question concerns the relatedness between the mechanisms of pathogenicity of curtobacteria and their taxonomy and phylogeny. The genomes of athovars ,21,22,23Curtobacterium taxonomy and genomic data. The abbreviations in the text below are as follows: C.—Curtobacterium, C. f.—Curtobacterium flaccumfaciens, C. fpf—flaccumfaciens pv. flaccumfaciens and C. f. pv.—Curtobacterium flaccumfaciens pathovars.In addition, this research concerns the development of existing diagnostic methods to better correspond to the Bacterial chromosomal and plasmid genomes were downloaded from the NCBI Genome database . ChromosA genome sequence comparison and a visualisation of the annotated plasmid genomes were made with EasyFig , applyinThe average nucleotide identity (ANI) was calculated using OrthoANIu . The datgyrB, parE, rpoA and rpoB were extracted from the annotated genomes and aligned with MAFFT [Genes sequences of the rRNAs, ribosomal proteins, th MAFFT  with L-Ith MAFFT . Phylogeth MAFFT  and RAxMth MAFFT  softwareth MAFFT  and Geneth MAFFT  nucleotiThe core genes’ concatenated alignment was obtained with the Panaroo pipeline  using PrProtein structure modelling was performed in two steps. First, the structure was predicted using AlphaFold 2.0  with defCurtobacterium from other genera with fairly good reliability. To develop primers, the 23S rRNA sequences of all the available Curtobacterium strains and a set of unrelated strains were aligned using MAFFT [Curtobacterium was manually determined. Oligonucleotides for the amplification of this region were optimised using Primer3Plus [The phylogenetic analysis showed that the 23S rRNA sequence could be used to differentiate ng MAFFT . Then, fmer3Plus .Curtobacterium [A total of 81 bacterial strains, listed in acterium . The remacterium ,56.Routine cultivation of the strains was carried out on YD medium  at a temperature of 28 °C. Long-term storage of the strains was carried out at −80 °C in 30% glycerol.A GeneJET Genomic DNA Purification Kit  was used to isolate genomic DNA following the manufacturer’s instructions. The concentration of purified DNA was measured using a NanoProteometer N60 spectrophotometer . On average, the DNA amount in the samples was ~20 ng.Conventional PCR was performed using a BioRad T100 ThermalCycler  in a volume of 25 μL. Each reaction contained five μL of ScreenMix , 0.3 mM of each primer, and 20 ng of template DNA.The thermal cycling mode was 94 °C for 300 s, followed by 30 cycles of 94 °C for 30 s, 65 °C for 10 s, and 72 °C for 20 s, with a circuit extension at 72 °C for 5 min. PCR products were visualised by electrophoresis (1.5% agarose gel with the addition of ethidium bromide and 1× TAE buffer). A 1-kb ladder length marker (Evrogen) was used to determine the size of the DNA fragments.To construct a test plasmid, the target sequence was first amplified in the Cff VKM Ac-1923 (← DSM 20129) strain. After the resulting PCR, the product was purified using the Qiagen MinElute PCR Purification kit  and ligated into the pDrive vector (Qiagen) using the Qiagen PCR Cloning kit. The correctness of the insertion was checked using Sanger sequencing.PCR was performed in a final volume of 10 μL. The reaction mixture included 2 μL of Evrogen qPCRmix-HS SYBR, 0.3 mM of each primer, and 20 ng of template DNA. The final concentration of dNTP was 0.12 mM, and the concentration of magnesium was 3 mM. Thermal cycling was performed on a LightCycler 96  in the following mode: 94 °C for 300 s, then 45 cycles of 94 °C for 10 s, 65 °C for 10 s, and 72 °C for 10 s. All experiments were carried out in duplicate per run and repeated twice. Thus, there were four technical replicates. The processing of the amplification curves and calculation of the threshold cycles were carried out using Roche software. Reactions with water were used as a negative control and with a test plasmid as a positive control.For the experiment to determine the detection sensitivity, tenfold dilutions of the test plasmid and genomic DNA Ac-1923 (← DSM 20129) were prepared. PCR was performed with each dilution in the same way as described above.Curtobacterium, including two metagenome assemblies. Most of the strains were unclassified on the species level; 35 strains were classified as Curtobacterium flaccumfaciens, including 27 pathovars; and seven strains were classified as representatives of seven other Curtobacterium species . The preliminary analysis demonstrated that one of these strains, Curtobacterium S6, had the lowest average nucleotide identity (ANI) value, i.e., about 69% of all the other Curtobacterium strains. The phylogenetic analysis using 16S rRNA and 23S rRNA sequences of the closest homologues belonging to different Microbacteriaceae genera placed strain S6-1 distantly from the Curtobacterium clade (data not shown). Seemingly, this strain has been misclassified and might possibly represent a novel genus. Thus, only 190 genomes can be considered as Curtobacterium spp.  can assist taxonomy ,58,59. TCurtobacterium genomes . According to the analysis of 3500 genomes representing type strains of species from >850 bacterial or archaeal genera [Interestingly, the ANI value of  75–76%, , was alml genera , the ANICurtobacterium are located in one operon, together with 5S rRNA, and are separated by internal transcribed spacer (ITS) regions. The majority of 16 complete Curtobacterium genomes contain three rRNA operons, and several complete genomes contain four copies of rRNA operons. In the latter case, two operons are located next to each other and are separated by the 1300–2200 base pairs (bp) region and are located in one direction. The average length of Curtobacterium 16S rRNA is about 1530 bases, and the average length of 23S rRNA is about 3120 nucleotides.Genomic loci encoding ribosomal RNAs are often used for evolutionary phylogenetic analysis and species delineation ,71,72. AThe copies of rRNA genes belonging to the same genome are often not identical. To conduct the phylogenetic analysis, the rRNA gene, which has an identical copy (copies), was chosen. If all the sequences were different and shared the same, or the adjacent, clade, the sequences with the least sum of branch lengths to the root of draft trees were chosen for the final tree.C. fpf CFBP 3423 was found to be identical to Moraxella osloensis YV1 16S rRNA, which could have been the result of sample contamination and assembly error. Furthermore, this gene was not located in a common operon, together with the 23S and 5S rRNA genes. One of the 23S sequences of C. sp. HSID17257 was 100% identical to a Cryptococcus neoformans complex 25S rRNA and showed a high level of similarity with rRNA genes found in plasmids  and several bacteria.The 16S sequence of Curtobacterium strains  did not improve the robustness of inference and resulted in a similar consensus tree topology. This can be due to the low difference between the 16S rRNA sequences—the pairwise identity  of the 16S rRNA alignment was as high as 99.4%.The results of phylogenetic analysis of 16S rRNA genes belonging to 185  strains  raised dCurtobacterium strains , topoisomerase IV subunit B (parE) evolutionarily related to gyrase B, DNA-directed RNA polymerase subunit α (rpoA), and DNA-directed RNA polymerase subunit β (rpoB). All the genes were found in single copies in almost all genome assemblies.Conservative genes coding for proteins, mainly related to DNA processing  and their concatenation, can be efficiently used for high-resolution phylogenetic analysis ,77,78,79parE encoding topoisomerase IV subunit B demonstrated good phylogenetic potential resolving the tree topology slightly better, and with a little better bootstrap support, than the gyrB, rpoA, and rpoB analyses, presumably because of the greater divergence of parE. The pairwise identities of the alignments were 91.4% for gyrB (189 sequences), 89.2% for parE (189 sequences), 96.6% for rpoA (189 sequences), and 95.7% for rpoB (188 sequences). It appears that a low number of variable sites hampers the fidelity and informativeness of the phylogeny.The phylogenetic trees of all the genes listed above  featuredgyrB, parE, rpoA, and rpoB and demonstrated a higher resolution and better bootstrap support than the single genes phylogenies and, moreover, the rRNA phylogenies. The topologies of the MLPA trees and the compositions of the clades supported with high bootstrap values were close to those of the ANI tree   employedANI tree . This coThe phylogenetic analysis based on the amino acid and nucleic acid sequences of ribosomal proteins (r-proteins) was shown to contain a reliable phylogenetic signal at a wide range of taxonomic depths, which was not significantly affected by mutational saturation or lateral gene transfer ,81,82,83rplA-rplF, rplI-rplP, rplR-rplY, rpmA, rpmB, rpmD, rpmE2, rpmG2, rpmH-rpmJ, rpsA-rpsD, rpsG-rpsO, rpsQ, rpsS, and rpsT. The total length of the concatenated alignment was 21,202 bases; the pairwise identity was 92.4%.In this research, the concatenated nucleotide sequences of r-proteins extracted from reannotated genomes were used. The list of r-proteins genes used for concatenation comprised 46 genes: C. flaccumfaciens, was very close to the ANI tree , as well strains C. sp. VKM Ac-1376 (Species 4) and C. sp. YC1 (Species 5)  B. The prC. f. pv. oortii CFBP 3400 and 30 unclassified Curtobacterium strains.The proposed genomospecies 6–10 constitute two related clades of phylogenetic trees  and adjaC. pusillum, the strain labelled as C. flaccumfaciens JUb65, and ten unclassified Curtobacterium strains. Taking into account the genetic distances and phylogenetic data, the strains, currently classified as C. pusillum, should be assigned to three distinct species.The proposed genomospecies 11–20 belong to a single clade of phylogenetic trees  and haveC. luteum. One strain is unclassified, and one strain represents a metagenome assembly. It appears that the strains, currently classified as C. luteum, should be assigned to two different species.The phylogenetic trees’ clade and ANI matrix cluster E, compriC. albidum, C. citreum, C. oceanosedimentum, an unclassified draft genome, and a metagenome assembly, constitute a clade on phylogenetic trees . The genomes of the plasmids with a low GC contain multiple repeats, CTTT, CCTTTT, and similar, with the overweight of the thymine residues.As of October 2021, there were ten database . They coClavibacter sepedonicus strain ATCC33113 [The plasmids pCff1 and pCff113 attract special interest because of virulence proteins reported to be encoded in their genomes . These pTCC33113 .Curtobacterium trypsin proteases were found in pathovars of Xanthomonas campestris and Xanthomonas citri. Homologues of several supposed virulence genes were also found in phytopathogenic Clavibacter, Dickeya, and other phytopathogenic bacteria. Distant homologues of pectate lyase were found to be encoded by the genomes of phytopathogenic nematodes.The genome of pCff1 was reannotated using Prokka  and a thThe pCff1 genome also contains the genes of conjugation apparatus, a toxin–antitoxin system, DNA repair and restriction, and several transposases.Curtobacterium strains with confirmed pathogenicity, Curtobacterium sp. PhB130 and PhB136 chromosomes, and Curtobacterium plasmids pCff113 and pCff119 , pCM2 , and pVQ28-1 .The sequence search on pCff1 demonstrated the presence of extended homologous regions in chromosomes of 13  pCff119 . In addiBacillus phage φ29 [−5); an unusual lemon-shaped archaeal virus Salterprovirus His1 ; Streptococcus phage Cp-1 ; human adenovirus C ; and other bacterial, archaeal, and eukaryotic viruses. Significant similarities between phage φ29 and pCff1 gp137 were also shown by the I-TASSER homology modelling server [Actinomycete hypothetical proteins.HMM–HMM motif comparison found meaningful similarities between gene product 137 of plasmid pCff1 and DNA polymerases of hage φ29   (No. 1–22) and eight Curtobacterium strains isolated by the authors from infected plants (No. 23–30), as well as 15 strains isolated according to the same protocol but which turned out to be representatives of other genera (No. 31–45). Additionally, a set of strains isolated from rotting plants was tested to confirm the absence of false-positive amplification with common phytopathogenic bacteria such as Clavibacter, Pseudomonas, Xanthomonas, Pectobacterium, Dickeya, and others (No. 46–81). To check the correctness of the amplification, the PCR product was sequenced for five random samples.The set of strains included 22 Curtobacterium strains, while no amplification was observed for non-Curtobacterium isolates.The qPCR results are shown in Curtobacterium isolates from other microbiota found in infected plants, including closely related Microbacteria.According to the results obtained, amplification proceeded quite selectively, making it possible to differentiate C. flaccumfaciens Ac-1923 (← DSM 20129) strain, serial tenfold dilutions and qPCR were conducted. The amplification curves for the plasmid experiment are shown in Additionally, for the qPCR detection system, the efficiency and sensitivity of the detection were evaluated. For greater accuracy in calculating the number of copies in the medium, a test plasmid was constructed. For this purpose, an amplified fragment of the target 23S rRNA region was ligated to the pDrive vector. With the resulting plasmid and the genomic DNA of the 3 cfu/mL, which is a normal sensitivity for such detection systems.The Cq values obtained made it possible to plot a graph of the dependence of the threshold cycle on the logarithm of the DNA concentration per reaction. The data obtained are presented in Curtobacterium (family Microbacteriaceae) was defined by Yamada and Komagata in 1972 for a group of motile Brevibacteria [C. albidum (Komagata and Iizuka 1964). Yamada and Komagata 1972 homotypic synonym: Brevibacterium albidum Komagata and Iizuka 1964 [uka 1964 ;C. ammoniigenes ;C. citreum (Komagata and Iizuka 1964). Yamada and Komagata 1972 [ata 1972 ;C. flaccumfaciens (Hedges 1922). Collins and Jones described this species as a plant pathogen with six recognised pathovars [athovars ;C. herbarum ;C. luteum (Komagata and Iizuka 1964). Yamada and Komagata 1972 [ata 1972 ;C. oceanosedimentum—approved as a homotypic synonym: Flavobacterium oceanosedimentum Carty and Litchfield 1978 [eld 1978 ;C. pusillum (Iizuka and Komagata 1965). Yamada and Komagata 1972 [ata 1972 .The genus C. glycinis from Glycine max, C. gossypii from Gossypium hirsutum, and C. oryzae sp. from Oryza sativa [Other species have been proposed but not yet approved, such as a sativa .Curtobacterium spp. available at the NCBI Genome database were analysed using different bioinformatic methods. Deposited complete and draft genomes include 35 strains classified as C. flaccumfaciens, only 27 of which are identified as C. flaccumfaciens pathovars with confirmed virulence to a certain host plant. Seven genomes were classified as being representatives of other Curtobacterium species: C. albidum, C. ammoniigenes, C. citreum, C. herbarum, C. luteum, C. oceanosedimentum, and C. pusillum (Curtobacterium spp. have not yet been identified to a certain species. The relatively large number of sequenced genomes of C. flaccumfaciens (18.4%) can be explained by the status of C. fpf as an emerging and regulated plant pathogen that is spreading rapidly worldwide and that occurs worldwide in legume-producing countries.In the current work, 190 genomes of pusillum . Thus, 7Curtobacterium. It is probable that this limitation is the major reason for the uncertain taxonomic positioning of 148 analysed genomes.The results of a phylogenetic analysis using ANI and conventional 16S rRNA and 23S rRNA genes showed limited applicability to the definition of species within the genus gyrB, parE, rpoA, and rpoB) produced phylogenetic trees with significantly higher resolution and bootstrap supporting values than the rRNA phylogenetic analysis. The topology of these trees  extracted from reannotated genomes was used. The resulting best-scoring phylogenetic tree obtained with RAxML  demonstrANI tree .Curtobacterium with 51 potential genomospecies can be proposed  and isolated from host plants over a period of 77 years from North America, South America, Europe, and Asia. The C. flaccumfaciens strains were grouped into three main clusters. Colony colour aside, the three clusters did not correlate with pathovar affiliation, isolation date, geographical location, or isolation host. Thus, the pv. flaccumfaciens strains, pathogenic on beans, were scattered among the three clusters as diverse as other Curtobacterium species. Strains from the same pathovars (hosts of isolation) were placed in different clusters, irrespective of their geographical origin [MLSA based on housekeeping genes recA, gyrB, ppK, atpD, dnaK, and rpoB was used to investigate the phylogenetic relationships among 84 strains of C. flaccumfaciens, which were distributed among four pathovars (l origin .Curtobacterium spp. strains with glycosyl hydrolases (GHs) and, thus, with the potential for being degraders of cellulose and other polysaccharides, were not phylogenetically related to the type strain of C. flaccumfaciens, because they shared only 85% ANI. Furthermore, strains GD1, BH-2-1-1, MCBA15_013, and YR515 probably belong to separate species that are currently unnamed.A core genome phylogeny of 50 genomes  providedC. flaccumfaciens in the literature and found the genetic diversity within the group to be greater than had so far been described. Only two strains, MMLR14-002 and MMLR14-014, were identified as being members of C. flaccumfaciens, while the remaining strains needed to be reclassified as novel taxa.Chen et al.  evaluateatpD, gyrB, ppk, recA, and rpoB), revealed that the three strains, MCBA15-007, MEB126, and UCD-AKU, were phylogenetically closely related to the type strain of the poinsettia (Euphorbia pulcherrima) pathogen (pv. poinsettiae) ICMP 2566T, while the strains MCBA15-005 and UNCCL17 were phylogenetically closely related to the type strain of the sugar beet pathogen (pv. betae) ICMP 2594T [C. flaccumfaciens must be attributed to different species, reinforcing the need to reconsider the taxonomy of phytopathogenic members of the species. Thus, previous evaluations of Curtobacterium strains using genome sequences or MLST/MLSA have illustrated the presence of strain groups that are different enough from type strains of the eight validated species for new unnamed species to be claimed.Recently, MLST, using the sequences of five housekeeping genes . Hence, the area of genetic discontinuity may vary, depending on the taxa considered and their unique ecophysiologic and evolutionary characteristics, and 95% ANI appears to be the genetic level that distinguishes most natural discrete populations and named species, but not necessarily all.”In answer to the critics of the general concept of Jain et al. , the autIt was concluded that, “for these reasons, taxon descriptions should not be based on a single metric or threshold but the careful investigation of ecological and functional data together with genetic relatedness ” .Curtobacterium and Clavibacter strains appears to be related to their virulence. The important role of plasmids has been shown for many phytopathogenic bacteria [The occurrence of pCff1-related plasmids in the pathogenic strains of bacteria ,104,105.bacteria ,107. HowClaviceps purpurea [Discovery of the gene that encodes the protein structurally similar to viral DNA polymerase I (which belongs to family B DNA polymerases) raises important questions about the origin and evolution of pCff1-related plasmids. A plasmid-encoded DNA polymerase is an unusual event for bacteria and archaea, but several incidents, including actinomycetes, have been reported ,111,112.purpurea . Additiopurpurea . It is nThe homologues of pCff1 putative DNA polymerase have been detected only in chromosomal and plasmid genomes of actinomycetes. The relatedness of pCff1 putative DNA polymerase with the DNA polymerases of bacteriophages and archaeal and eukaryotic viruses have been revealed with the HMM–HMM motif comparison and structure modelling. These analyses have demonstrated that pCff1 putative DNA polymerase is more similar to the phage DNA polymerase. It is possible to hypothesise about the comparatively ancient origin and divergence of this protein. It is necessary to study the functioning of this enzyme experimentally. It might be suggested that the pCff1 putative DNA polymerase I could participate in the initial stages of replication, as has been reported in the context of some other plasmids ,122,123.Curtobacterium. The set makes it possible to differentiate Curtobacterium from representatives of other microbiota of an infected plant, which is an urgent applied problem. Primers for the species-specific detection of Cff have previously been described in the literature [The current study has seen, for the first time, the development of a genus-specific primer set for the diagnosis of terature , but theterature .The resulting test system can become a valuable and easy-to-use tool for rapidly assessing the strains isolated from a diseased plant. In addition, the sensitivity of detection should allow the use of this method for assessing plant extracts for the presence of a pathogen, even in asymptomatic plants.Curtobacterium. Based on these data, it is possible to discuss the delineation of up to several dozen genomospecies. A monophyletic group of 53 strains can be assigned to the species of Curtobacterium flaccumfaciens, which can include both pathogenic and non-pathogenic strains. The pathogenicity of Curtobacterium and other Microbacteriaceae can be related to a group of plasmids carrying virulence factors and featuring the presence of a gene distantly related to viral DNA polymerase I. The recent increase in genomic data challenges widespread diagnostic methods. The presented genus-specific PCR diagnostic kit developed and tested in the present work may serve as a good complementary tool for further studies of Curtobacterium strainsANI comparisons and the phylogenetic analysis of ribosomal and core proteins testify to the necessity for global taxonomic revisions within the genus of


Concrete, the most common material in the building industry, involves the use of mineral aggregates that represent an exhaustible resource, despite their large availability. For a series of applications, these mineral aggregates can be replaced by vegetal ones, which represent an easy renewable natural resource. In this study, two types of vegetal raw materials, namely sunflower stalks and corn cobs, were used in developing 10 compositions of ecological microconcrete, with different percentages involved: 20%, 35%, 50%, 65% and 80%; they were analyzed from the perspectives of density, compressive strength, splitting tensile strength, resistance to repeated freeze-thaw cycles, modulus of elasticity and thermal conductivity. The results revealed that the microconcretes with sunflower stalks registered slightly higher densities and better results regarding the compressive strength, splitting tensile strength, modulus of elasticity, and freeze-thaw resistance than those with corn cobs. Lightweight concrete is obtained when more than 50% replacement rates of the mineral aggregates are used. In 2018, they were responsible for 39% of the total CO2 emissions and for 36% of the global energy consumption [Buildings and construction represent high-demand sectors in terms of energy consumption and CO2 for each ton of cement produced [Concrete represents the most used building material all over the world but it is a nonecological material by definition, if we take into account the amount of exhaustible raw materials used for its production, and the energy consumption involved in obtaining Portland cement. To produce one ton of Portland cement, around 1.5 tons of raw materials are used  . Cement produced .Concrete greening methods studied so far include the substitution of cement, as its production determines an energy consumption of about 92% of the total energy , with vaNumerous studies have been carried out on replacing the mineral aggregates in concrete. They have used various variants of substitute materials for mineral aggregates, natural or industrial: bamboo , sugarcaHelianthus annuus) is a widespread agricultural plant in South America, Southern Europe, European Russia and South Africa [Zea mays) is a widely used agricultural plant in Eastern Europe, the United States, China, South Africa, cultivated in 2019 on more than 189,000 ha globally [Among vegetal aggregates suitable for use in concrete are corn cobs and sunflower stalks, given their wide global distribution, annual regeneration nature and the encouraging results of studies carried out so far on these plants ,29,30,31h Africa , grown oh Africa . Maize  and a peripheral woody part (bark) , very si 2 kg/m3 . These v 2 kg/m3 . The the 2 kg/m3 . In rese 2 kg/m3  on the a 2 kg/m3  developeThe aim of this research is to realize a comparison between concrete with aggregates from corn cobs and concrete with aggregates from sunflower stalks. The method applied for obtaining and treating the aggregates was approached similarly to that presented in detail in a previous study . A microThe present study involved the development of microconcrete recipes with vegetal aggregates from corn cobs and sunflower stalks, respectively. As a starting point, we used a reference concrete (RC) made with a conventional microconcrete recipe of the C25/30 strength class, with a maximum aggregate size of 8 mm, according to NE 012/1-2007 . This cl-aggregates: natural sand with diameter of up to 4 mm, and river gravel with diameter of 4–8 mm;-cement type CEM II/A-LL 42.5R MPa, produced in Romania;-polycarboxylate ether superplasticizer additive;-rhodanide-based accelerator additive, to speed up the early strength development of the vegetal concretes , so wateRC was made using the following materials:The water/cement ratio used was 0.50.In the RC recipe, the mineral aggregates were replaced with vegetal ones in volumetric proportions of 20%, 35%, 50%, 65% and 80%. Thus, we obtained five concrete compositions with corn cob aggregates, and five concrete compositions with sunflower aggregates. The vegetal aggregates were in granular form with variable diameter smaller than 7 mm in the case of corn cobs, and chips and fibers smaller than 25 mm length combined with granules with variable diameter of up to 7 mm in the case of sunflower stalks ; they weThe 10 concrete recipes with vegetal aggregates were noted as SFCXX or CCCXX, XX denoting the volume percentage  of sunflower aggregates or corn cob aggregates, respectively.In The density of the hardened concrete was determined 28 days after pouring, according to the methodology of EN 12390-7:2019 . The denThe concrete specimens were tested in monoaxial compression, with a hydraulic concrete press carrying a load evenly distributed on the surface of the specimen, according to EN 12390-3:2019 . The speThe determination of splitting tensile strength was performed on cylinders with diameter of 100 mm and length of 200 mm. This test consisted in compressing a specimen after two diametrically opposed generators .The tests were performed according to EN 12390-6:2009 . A hydraThe freeze-thaw resistance test  was perfAfter completing 50 cycles of 4 h of freezing and 4 h of thawing, the specimens were compression tested, and the loss of compressive strength was determined.For determination of the modulus of elasticity to compression of the concrete developed in this research, method 2 of EN 13412:2006  was appl2 and one third of the compressive strength of the specimen, determined according to EN 12390-3:2019 [The modulus of elasticity is determined by measuring the change in tension in the concrete test specimen at loading, to produce a pressure (force) between 0.5 N/mm0-3:2019 .The thermal conductivity was determined according to a method provided in SR EN 12667:2009  on the tIn the results analysis, we realized an interpretation of the individual values of the vegetal concretes ; the following variants, SFC35, SFC50, SFC65, and SFC80, recorded approximately similar values of this parameter, with small variations between 0.80–0.99 N/mm2.In the case of concrete with sunflower aggregates, there was also a decrease in splitting tensile strength as the percentage of plant material increased . The incIn a comparative analysis, according to RC registered a modulus of elasticity of 35,809.80. According to In terms of compressive strength, after being subjected to 50 freeze-thaw cycles, RC registered a decrease of 12.74%. Concrete with corn cob aggregates recorded decreases to a greater extent than RC . Thus, gIn comparative analysis , and above those that implied broken corn cobs, not shredded. The compressive strength of CCC80 was above other studies on concrete with similar corn cob content. SFC80 recorded bigger or similar values for compressive strength and better thermal conductivity than similar studied materials.As for advantages of sunflower stalks and corn cobs in concrete composition, they had a significant influence in reducing density and thermal conductivity, in addition to their ecological and sustainable character. A concrete with lower density and a more elastic character could lead to a better behavior of the buildings in case of seismic activity, since it would lower overall weight if it is used in nonstructural applications, such as screeds, plaster, partition walls or in masonry blocks form. As screeds, adding their thermal insulation characteristics, a reduction of the thermal bridges at the slab level can be obtained.SFC and CCC can improve a building’s sustainability by diminishing their total carbon footprint through the use of easy-to-obtain, renewable raw materials instead of exhaustible ones.


No muscle pain or rhabdomyolysis was reported. Results from our study highlight that training based on short-duration high-intensity exercise improves overall metabolism and aerobic fitness, thus being feasible, at least in a case of CPTII deficiency.In the mild subtype of inherited carnitine palmitoyltransferase II (CPTII) deficiency, muscular mitochondrial fatty acid β-oxidation is impaired. In this condition, interventions involve daily dietary restriction of fats and increase of carbohydrates, whereas physical exercise is commonly contraindicated due to the risk of muscle pain and rhabdomyolysis. We present the case of a 14-year-old female with CPTII deficiency who underwent a 1-h session of unsupervised exercise training for 6 months, 3 days per week, including interval and resistance exercises, after diet assessment and correction. Before and after intervention, the resting metabolic rate (RMR) and respiratory quotient (RQ) were measured by indirect calorimetry, and a cardiopulmonary exercise test  was performed. Interval training consisted of a 1 min run and a 5 min walk (for 15 min progressively increased to 30 min). During these efforts, the heart rate was maintained over 70% HR max corresponding to respiratory exchange ratio (RER) of 0.98. Resistance training included upper/lower split workouts . Blood CK was checked before and 36 h after two training sessions chosen randomly without significant difference. After training, RMR increased (+8.1%) and RQ lowered into the physiological range (from 1.0 to 0.85). CPET highlighted an increase of peak power output (+16.7%), aerobic performance (VO The carnitine palmitoyltransferase (CPT) system includes two enzymes, CPTI and CPTII, located in the outer and inner mitochondrial membranes, respectively, and involved in the transportation of long-chain fatty acids (LCFA) into the mitochondria .CPTII deficiency is the most frequent muscular inherited disorder of the muscle fatty acid metabolism resulting in reduced LCFA oxidation as an energy source, both at rest and during muscular activities. CPTII deficiency leads to a spectrum of clinical conditions ranging from a lethal neonatal disorder, a severe infantile hepatocardiomuscular disorder and a mild myopathic form . Most ofClinical symptoms of the disease occur more frequently during infancy (1–12 years old) than in adolescence (13–22 years old) and adulthood (>22 years old) . Muscle The biochemical consequences of the disease-causing mutations are still debated. Previous studies highlighted that the enzyme activity ranges from undetectable  up to noIn CPTII deficiency, to prevent symptoms and rhabdomyolysis, the current interventions involve a daily dietary restriction of fats and increased carbohydrate intake. Glucose infusion is adopted whenever the patient has a fever or infections that raise the metabolic demand. The suggested nutritional regimen results in a short-term increase of the exercise tolerance but recurrent rhabdomyolysis has been also observed in these cases . TherefoDespite these premises, we speculate that a specific type of training, based on short duration contractions and resistance exercises, can be beneficial also for CPTII deficient patients. In fact, it is widely known that the energy sources used during exercise strictly depend on the duration and percentage of the maximal sustainable load . In partWe report here the case of a 14-year-old girl complaining of lower-limb weakness and myalgia following physical exercise at school. The patient and her parents gave their written consent to participate in the study and for the data to be published after a thorough explanation about the purpose of the study, which was approved by the local institutional ethics committee.In the clinical and biochemical workout hyperCKemia was found  without any other neurological signs or symptoms. The girl’s pre- and perinatal history of was normal, as was the family history. The parents were not consanguineous. In the recent history of the girl a couple of similar episodes  were reported, both following dancing and skiing without dark urine. Both of the episodes resolved in about 24 h. HyperCKemia was confirmed 1 month later, even without reported exercise in the previous hours . Neurological examination was normal. Biochemical study in the family showed high CK in her apparently healthy brother, so a Multiplex Ligation-dependent Probe Amplification (MLPA) analysis of the Dystrophin gene and Dried Blood Spot (DBS) enzyme assay for Pompe disease were carried out, both with negative results. Muscle magnetic resonance imaging was normal: no remarkable signs of muscle disease were found. Spirometry and cardiologic examination did not show any abnormalities. Then a muscle biopsy was performed: it showed slight morphological alterations, possible expression of a previous metabolic rhabdomyolysis, or, alternatively, a paucisymptomatic muscular dystrophy. No increased content of glycogen and lipids was seen; myophosphorylase and myoadenylate deaminase activity, as well as the mitochondrial enzyme activity, were normal. Anti-dystrophin antibodies displayed a normal pattern. Muscle biopsy and clinical features suggested a lipid metabolic disorder, and the dosage of CPTII was recommended: enzyme assessment showed a reduced activity. Then, genetic analysis confirmed the diagnosis: deficiency of CPTII, with a mutation in the heterozygosis of p. S113L . Subsequent genetic analysis confirmed that the brother carried the same mutation.After diagnosis, the use of a weekly food diary template highlighted consumption of fats as well as processed foods. A personalized low-fat normocaloric diet of 2,000 kcal/day was achieved based on the estimated calorie needs per day, by age, gender, and physical activity level . A nutri2), carbon dioxide production (VCO2), and respiratory quotient (RQ = VCO2/VO2 in steady state conditions), considered an index of which type of fuel is mainly metabolized during the test   was 6.5%. Initial RMR was 1,597 kcal/day, with an RQ of 1.07 , highlighting a high percentage of carbohydrates utilization (92%).After 3 months of adherence to the new diet regimen, the resting metabolic rate (RMR) was measured using indirect calorimetry technique  under optimal environmental conditions (23°C at 8:00 a.m. in fasting conditions). RMR provides information in the form of oxygen consumption (VO.7 fats) . Gas exc2 and VCO2 output were measured using the breath-by-breath method. The test was performed until exhaustion, with the achievement of a respiratory exchange ratio (RER) of 1.16 . The maximal power reached was 120 watts. VO2peak was 1,693 ml/min corresponding to 21.7 ml/min/kg . The heart rate peak (HRpeak) was 153 bpm, 75% of the predicted HRpeak.Further, a cardiopulmonary exercise test (CPET) was performed on a cycloergometer , with the patient wearing a two-way breathing mask covering the nose and mouth , connected to a gas analyzer . CPET was performed with an incremental technique , and VO2 was below 30, but from the metabolic impairment.Overall CPET underlined a very low oxygen uptake during the effort, not from respiratory deficiency, as VE/VCOAfter getting used to the proposed exercises, the patient was involved in an unsupervised 1 h exercise training session three times per week for 6 months. Each session included interval training and resistance exercise activity. The interval training consisted of a 1 min run and 5 min walk for 15 min progressively increased to 30 min (0° inclination). During the patient’s running, the heart rate, monitored with a cardiofrequenzimeter, was maintained over 70% HRmax (corresponding to a RER of 0.98 measured at the first CPET). Resistance training was based on three sets of eight repetitions with 2 min rest between sets. During training, upper/lower split workouts were carried out. Upper-body exercises included bench press, shoulder press, lat pulldown, dumbbell flies, and dumbbell pull-over and arm curls. Load was fixed at 6–7 of the 0–10 scale for resistance exercise  accordinBlood CK was checked before and 36 h after two training sessions chosen randomly with the following results: pre-313 U/L, post-314 U/L; pre-439 U/L, post-242 U/L). Before, during, and after each session, the patient drank a total of 500 ml of a solution containing 22.4 g of carbohydrates, 3 g of creatine, 248 mg of sodium, and 146 mg of potassium .2peak, 8,3%) and anaerobic threshold (+5.7%), oxygen pulse (+4.5%) and longer isocapnic buffering period (IB) (+335%) (2 was still under 30 (25.3) underlining the absence of respiratory deficiency (Before and after training RMR and CPET results are reported in  (+335%) . VE/VCO2ficiency .This study demonstrated that under the regimen of a properly balanced diet based on a high consumption of carbohydrates and a low intake of fat, a training program with high intensity/low duration bouts of exercise is safe and associated with an overall improvement of body metabolism and aerobic fitness in a CPTII-deficiency patient.Initial RMR and cardiopulmonary tests revealed high carbohydrate utilization at rest and scarce aerobic fitness, below what was predicted for age and gender. Following exercise training combined with diet adjustment, RMR increased and RQ decreased toward normalization, suggesting an improvement of metabolic active muscles combined with the optimization of macronutrient utilization. In light of the genetic nature of the impaired fatty acids metabolisms, the observed change in RQ probably reflects an increased efficiency in the energy disposal and utilization of carbohydrates at rest and/or an amelioration of the residual fat utilization through alternative pathways .2peak, VO2 at the anaerobic threshold (AT), and oxygen pulse and isocapnic compensation. Given the results obtained, we can speculate that both muscular  and cardiovascular adaptations, including changes in cardiac contractility, blood volume, and cardiac output, may have contributed to the observed VO2peak improvement. This possibility is mirrored by concomitant changes in resting RQ, which may justify significant amelioration of the mitochondrial expression and/or function and increased oxygen pulse known to represent an indirect indicator of the cardiac stroke volume  deficiency , followie volume . Importae volume .2peak  into the muscle cell. Under resting conditions, Ca2+ cellular concentration should remain at nanomolar levels. Ca2+ would increase through cell activation and muscle contraction when ATP storage is depleted , activation of Na+-Ca2+ membrane exchanger, and rupture of the sarcolemma (2+ has been reported to increase the protease and lipase activities that determine sarcolemma damage. In a context in which lipid oxidation is not efficient, as in CPTII, an increased exercise-induced susceptibility to rhabdomyolysis is expected (Importantly, our results also indicated that the training protocol was safe, as neither significant post-exercise changes in blood CK nor muscle pain or rhabdomyolysis was experienced by the patient all along the experimental period. The main pathophysiological mechanisms involved in the progression of rhabdomyolysis include an increase of calcium (Cadepleted . Severalrcolemma . An increxpected . In factexpected . In our expected .The results of this study show that, after diet adjustment, a training protocol based on high intensity/low duration contractions is safe and may improve metabolism and aerobic fitness in patients affected by CPTII deficiency. Nevertheless, as a single case report, the results must be interpreted with caution and cannot be generalized to other patients, and future studies with proper statistical analysis are needed to confirm the findings. Moreover, we considered a young female patient, and we cannot exclude the possibility that different outcomes may be obtained from training male CPTII deficient subjects, who experience more severe and frequent clinical presentation of the disease .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 Institutional Review Board. Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin.MP analyzed and interpreted the data, wrote the first draft of the manuscript. EP provided medical care, testing, and medical tracking of the patient. LC supervised functional measurements and interpreted the data. SA analyzed and interpreted the data. MN provided nutritional assessment and correction. MB contributed to writing and revision of the manuscript for intellectual content. AB provided medical care, testing, and medical tracking of the patient, writing and revision of the manuscript for intellectual content. GD’A developed the conception of the study, interpreted the data, and wrote the first draft and the final version of 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.


Resin acids are antimicrobial and anti-inflammatory compounds derived from spruce and pine trees. In a previous study, in which a mixture of resin acids and fatty acids from pine and spruce trees was added to the feeds of broiler chickens, they grew heavier, and their footpads stayed in better condition. Here, three separate trials were conducted to study whether dietary resin acids without the fatty acids have the same effects on broiler chickens. Broiler chicken diets were amended with different dosages of a resin acid concentrate product. Bird weight gain increased, and the efficiency of broiler production was improved when the resin acid concentrate was added to the diet at 125–250 g/ton. The litter material of pens stayed drier in resin acid concentrate groups, which may indicate better intestinal condition of the birds. However, the footpad condition was unaffected by the experimental diets. The results suggest that the resin acid concentrate improves bird weight gain and the efficiency of broiler production. Therefore, the resin acid concentrate may be a promising feed additive for broiler chickens.Dietary coniferous resin acids have previously been suggested to support the intestinal integrity of broiler chickens by reducing mucosal collagen degradation. The present study examined the effects of resin acid concentrate (RAC) on broiler performance and litter quality. In trial 1, RAC was added to diets at 0, 125, 250, or 1250 g/ton, while in trials 2 and 3, RAC dosing was 0 or 175 g/ton. Bird weight, feed consumption, mortality, feed conversion ratio (FCR), European Efficiency Index (EEI), litter moisture, and footpad dermatitis (FPD) lesions were measured. In trial 1, RAC at 125 and 250 g/ton improved weight gain and EEI, while RAC at 1250 g/ton group did not differ from control. Feed consumption, FCR, FPD scores and mortality were similar in all treatments, but litter quality was improved by all doses of RAC. In trials 2 and 3, RAC increased the final weight of birds, improved FCR, EEI, and litter quality, but had no effects in other parameters. In summary, RAC at 125–250 g/ton improved bird performance and thus shows promise as a feed additive. The dryer litter in RAC treatments may suggest improved intestinal condition as a response to in-feed resin acids. During the past two decades, novel research methodologies have provided a lot of detailed information on the association of intestinal inflammation and suboptimal production performance in poultry and other farm animals ,2,3. GutPinus sylvestris L.) and Norway spruce (Picea abies L.) and which has since 2015 been marketed for farm animal feeds in some areas and countries such as the European Union, Norway, Switzerland, Turkey, Thailand, and Mexico. Amending diets with TOFA has improved the production performance of broiler chickens [Clostridium perfringens [A newcomer to the group of plant-based feed supplements is tall oil fatty acid (TOFA)  which cochickens ,10,11, tchickens , and sowchickens . In vitrfringens ,11,14, wfringens ,10. Favofringens  may partWhile the fatty acids of TOFA, mostly linoleic and oleic acids, are common in poultry feeds because of their presence in other feedstuffs, no history seems to exist for the intentional addition of purified coniferous resin acids to farm animal feeds. Resin acids, the main components of coniferous rosin, are hydrophobic diterpene carboxylic acids with antibacterial, antifungal, and anti-inflammatory activity ,16,17. AWhen purified natural mixture of coniferous resin acids was fed to broiler chickens at the dietary level of 200 g/ton for three weeks, the density of inflammation-associated T-cells in the duodenal mucosa was reduced, and the collagenolytic matrix metalloproteinase activity in the ileal mucosa was inhibited . These rThe hypothesis of the present study was that if resin acids have a positive effect on gut barrier and homeostasis, they may also improve the performance of broiler chickens, reduce the moisture of litter material in pens, and inhibit the development of FPD lesions. Three experiments were conducted including different doses of RAC in broiler feeds until 35 days of age. Bird performance, efficiency of broiler production, FPD incidence, and litter moisture were measured in all three trials. The dosing of RAC was from 125 to 1250 g/ton, the highest dosing giving information on the safety of the product to the birds.Pinus sylvestris L.) and Norway spruce (Picea abies L.), and it was produced by Forchem Ltd. . The analyzed total content of resin acids in RAC was 37.5% (±0.2%).The product RAC  contained 40.0% tall oil rosin, 60% of food grade whole grain wheat flour, and antioxidants BHT (175 ppm) and BHA (175 ppm) in the form of dry flour. Tall oil rosin was derived from the tree species Scots pine  in the amount of 1 mg/kg of diet. Moreover, all the premixes contained the enzymatic product RovabioR  and PhyzymeR XP . The birds received a basal diet (Control) or diets  supplemented with RAC at 125, 250, or 1250 kg/t . Selecting the dosing of RAC for was based on the assumption that resin acids are the active ingredient in the product and also in the existing TOFA product  [In the completely randomized trial 1, conducted from November–December 2019, newly hatched Ross 308 broilers, 840 in total, were randomly divided into four dietary treatments and each further into seven pens of 30 chickens. Complete formulations adjusted to the demands of fast-growing birds were used . The amon acids) ,11,12,13R, Elanco, Greenfield, IN, USA) were produced, one for trial 2 and the other for trial 3  per pen was calculated by dividing feed consumed (kg) by weight gain (kg) of the respective time period. Feed intake was corrected for mortality by deducting the estimated amount of feed intake of dead birds. Fattening efficiency was determined on the basis of the European Efficiency Index (EEI). To calculate it, the following data were used: length of fattening, feed intake, final bodyweight, and the survival rate in each group:For each pen, one foot per bird was systematically evaluated for macroscopic examination and scored in three classes of FPD on days 14, 28, and 35 as follows: 0 = No lesions; no or very small superficial lesions, slight discoloration on a limited area, mild hyperkeratosis; 1 = Mild lesion; discoloration of the foot pad, superficial lesions, dark papillae; 2 = Severe lesion; ulcers or scabs, signs of hemorrhages or swollen foot pads according to the classification of Ekstrand et al. [From all trials, litter quality (moisture) was measured per pen by electronic device  at days 14, 28, and 35 as a standard procedure at the experimental farm. Measurements of all observations, from five different sites per pen, were averaged per pen.t-test. For both trials, the results were characterized using the arithmetic mean (x), the standard error of the mean (SEM), and the level of significance (p). The computer program STATISTICA 12 was used for calculations.The pen was considered the experimental unit for statistical purposes. For trial 1, the results were analyzed statistically using one-way analysis of variance (ANOVA) and Duncan’s test. For trials 2 and 3, the results were analyzed statistically using Student’s The results of the analysis of nutritional components of the experimental diets are given in In all three trials, the general health of the birds was good during the experimental period, and no medication was used. Birds were growing at typical rate for the breed.p < 0.01). The final weight of birds in group RAC1250 was similar to weight of control birds. The total feed intake, mortality and FCR were similar in all dietary treatments, although in the second week, RAC at 125 and 250 g/ton tended to improve FCR. The EEI value was significantly improved in the RAC125 and RAC250 groups by 8.1% (p < 0.05) and 10.9% (p < 0.01), respectively. There was a significant positive effect of RAC addition at 250 and 1250 g/ton on litter quality on days 28 and 35, and for RAC1250, already on day 14 . Feed intake and mortality were similar for both groups. The use of RAC significantly improved EEI . Days 1–35 showed a trend for better FCR in RAC-treated chickens (p = 0.067). Litter quality was significantly improved by RAC175, compared with control, on day 14, but similar to control in days 28 and 35. Healthy feet with no FPD lesions were observed in all inspected birds, and thus no differences in the FPD score were found between groups (data not shown).All results of trial 2 are presented in p < 0.01). Feed intake and mortality were not affected by the dietary treatments. For the first four weeks, FCR was similar for both treatments, but for days 1–35, FCR was significantly better for RAC175 group than for the control group (p < 0.01). Dietary RAC significantly improved EEI . There was positive effect of RAC175 on litter quality on day 28 (p < 0.1) and day 35 (p < 0.05). The inspected birds had healthy feet with no FPD lesions, and thus, the treatment groups showed no differences in FPD scoring (data not shown).All results of trial 3 are presented in The performance and feed efficiency of broiler chickens is closely associated with their intestinal functions . EntericAccording to the feed analysis results, the quantity of basic nutritional components in experimental diets were at targeted levels in all three trials. The analysis of resin acids from feed samples confirmed that RAC was dosed to the experimental diets at acceptable accuracy, and none of the control diet samples contained resin acids. The discrepancy between calculated and analyzed resin acid concentrations is mainly due to the feed matrix interfering with the extraction of resin acids, leaving the error of margin of this analysis typically from 10 to 20%, although the possibility of other contributing factors, such as feed mixing and sampling, needs to be kept in mind.One of the main observations of the present study was the significant increase in the final weight of the chickens by dietary resin acids. On day 35 of all three trials, the weight of birds fed with RAC at 125, 175, or 250 g/ton of feed  was significantly higher than that of control birds, the magnitude of the effect being from 3.0% to 6.1%. The range of body weight improvement in the present study is well in line with two broiler studies, in which dietary TOFA, providing 45 g/ton or 67 g/ton resin acids, increased the final weight of broiler chickens by 3.3%  or 3.6% In all three trials, feed intake was similar for all dietary treatments, including the highest RAC dose of 1250 g/ton in trial 1. The result indicates that at doses of 125–1250 g/ton, RAC does not influence the palatability of broiler diets, and that it causes no such physiological effects which would reduce feed intake. In line with this observation, the studies describing the performance effects of TOFA on broiler chickens or turkeys have reported no effects on feed intake ,10,11,12p < 0.1) towards the same direction was seen in trial 2. The average FCR values of control groups were 1.53, 1.62, and 1.66 for trials 1, 2, and 3, respectively. It can be speculated that the good FCR in trial 1 did not leave much space for improvement. In any case, the ability of RAC to affect FCR in broiler chickens needs to be verified in future studies. For antibiotic growth promoters, an average improvement of FCR is 2.64% [Mortality was not affected by the dietary treatments. The birds were in normal condition in these trials and not affected by any infections or stress factors which would have increased the mortality over normal levels. The FCR values showed inconsistent results between the trials. Only in trial 3, the resin acid treatment significantly improved FCR for days 1–35, although a trend (is 2.64% .Another way to measure the efficiency of meat poultry production is the EEI, which takes into account broiler mortality, final weight, feed conversion, and duration of the rearing period. In all the three trials of the present study, amending the feeds with RAC at doses of 125–250 g/ton significantly improved EEI by 5.1–10.9%, compared with the respective control groups. The 250 g/ton dose of RAC showed the largest response to EEI. In comparison, TOFA dose providing 45 g of resin acids/ton of feed has been reported to result in 6.4% improvement of EEI [The motivation to measure litter moisture and footpad lesions arose from a study suggesting that dietary resin acids support gut barrier functions by reducing the inflammation-associated collagen degradation of intestinal mucosa . Gut leaIn contrast to the hypothesis, RAC amendment did not reduce FPD lesions in the present study, although it should be noted that birds in trials 2 and 3 did not suffer from FPD lesions at all. In a previous broiler study, TOFA, providing resin acids 45 g/ton, significantly reduced FPD scores by 18%, but the score of the control group was 91.14 . In 15-wThe 1250 g/ton RAC dosing was designed to give information on safety margins of this dietary additive. The intended level of usage of this feed additive lies between 125 and 250 g/ton, so the 1250 level represents a 5–10 × overdose to this range of levels. As resin acids are fat-soluble molecules not commonly found in feed ingredients, it is necessary to ensure their safety in diets of meat poultry when fed at practical levels. Although published information on the toxicity of resin acids on chickens does not exist, studies on laboratory rodents suggest quite low toxicity, as discussed by Apajalahti et al. . In broiThe effects of a novel resin acid-based product, RAC, on broiler chickens were studied in three separate 35-day trials. The positive effects of RAC on broiler performance were demonstrated here for the first time. Higher EEI as a response to dietary resin acids suggests improved profitability of broiler production at the dietary RAC levels of 125–250 g/ton. Resin acid treatments resulted in a reduced litter moisture, but no effects on FPD lesions were observed. The reduced litter moisture may suggest that resin acids support the intestinal condition and thus inhibit the risk for diarrhea. Overall, the results suggest that the resin acid concentrate is a promising feed additive for broiler chickens.


Neuropilins are transmembrane glycoproteins that play important roles in cardiovascular and neuronal development, as well as in immunological system regulations. NRP1 functions as a co-receptor, binding numerous ligands, such as SEMA 3 or VEGF and, by doing so, reinforcing their signaling pathways and can also interface with the cytoplasmic protein synectin. NRP1 is expressed in many cancers, such as brain cancers, and is associated with poor prognosis. The challenge today for patients with pediatric brain tumors is to improve their survival rate while minimizing the toxicity of current treatments. The aim of this review is to highlight the involvement of NRP1 in pediatric brain cancers, focusing essentially on the roles of NRP1 in cancer stem cells and in the regulation of the immune system. For this purpose, recent literature and tumor databases were analyzed to show correlations between NRP1 and CD15 (a stem cancer cells marker), and between NRP1 and PDL1, for various pediatric brain tumors, such as high- and low-grade gliomas, medulloblastomas, and ependymomas. Finally, this review suggests a relevant role for NRP1 in pediatric brain tumors progression and identifies it as a potential diagnostic or therapeutic target to improve survival and life quality of these young patients. Neuropilins (NRPs) are transmembrane glycoproteins found in all vertebrates and are highly conserved across species. Two homologous NRPs isoforms have been identified, namely neuropilin-1 (NRP1) and neuropilin-2 (NRP2). NRPs are non-tyrosine kinase surface proteins that play important roles in neuronal development, cardiovascular development, and in the immune system . NRPs arIn children, primary tumors of the central nervous system (CNS) are the second most common cause of cancer after leukemia, accounting for approximately one fourth of all childhood cancers. There are approximately 5,000 new cases diagnosed each year in the United States among children younger than 18 years . ActuallAlthough the roles of NRP1 in tumor angiogenesis, tumor microenvironment, as well as its possible targeting in tumor progression, have recently been reviewed elsewhere –20, in tGliomas are the most frequently pediatric brain tumors and represent 25% of all brain tumors. They are heterogeneous and are divided into two main categories, low- and high-grade tumors, according to their aggressiveness and their prognosis. Low-grade gliomas (LGGs), such as pilocytic astrocytoma (PA), are usually curable while high-grade gliomas (HGGs), such as diffuse intrinsic pontine glioma (DIPG), are consistently fatal.LGGs, classified by the World Health Organization (WHO) as grade I or II, comprise several subgroups, including PA, pleomorphic xanthoastrocytomas (PXA), pilomyxoid astrocytomas (PMA), subependymal giant cell astrocytomas (SEGA), low-grade fibrillary astrocytomas, and diffuse astrocytomas. The most common LGG diagnosed in children aged 0 to 14 years is the PA, which accounts for 85% of all LGGs . Five-yeMedulloblastoma (MB) is the most common malignant brain tumor of the posterior fossa in pediatric, accounting for 15% to 20% of all childhood brain tumors. The survival rate of patients with embryonal tumors of the central nervous system, such as MB, is 54% at 5 years. The prognosis is significantly worse before the age of 1 year: 62% survival rate at 5 years, compared to 81% for children aged 10 to 14 years , 37. BecEpendymomas (EPNs) are the third cause of malignant brain tumors in children. These tumors affect mainly the children younger than 5 years. EPNs are usually located in the posterior fossa (70% of cases), but they can develop anywhere within the central nervous system. The histological grade score of the tumor is not reproducible and has no prognostic impact. Recent advances in molecular biology confirm that, depending on their localization , EPNs are different diseases. These tumors have a worse prognosis in children than in adults , 42. EPNThere is a wide diversity of pediatric brain tumors that are very rare, accounting for less than 30% of all tumors. Some of them, such as the choroid plexus papillomas, a benign (WHO grade I) neuroepithelial intraventricular tumor, have an excellent prognosis after complete surgical resection when they occur in children. Other tumors, such as craniopharyngiomas, arising from the sellar region, also have a very good overall survival after complete resection, but relapses may later occur that necessitate a complementary treatment by radiotherapy. The excellent overall survival of these tumors is unfortunately impaired by endocrine, and often visual, sequelae.Other embryonal tumors of the CNS, like atypical teratoid and rhabdoid tumors, and embryonal tumors with multilayered rosettes, are highly aggressive, poorly differentiated, and occur predominately in young children. Their treatment is therefore very challenging, and their prognosis is still grim.Unfortunately, despite the progress made in recent decades in the management of these cancer types, the effectiveness is still suboptimal and sequelae are frequent. Indeed, chemotherapy has been shown to provide significant toxicities and adverse side effects , 47. SimNRPs are transmembrane glycoproteins specific to vertebrates. There are two members in this family of proteins, NRP1 and NRP2, which are coded by two different genes located on two chromosomes (10p12 for NRP1 and 2q34 for NRP2). NRPs play an important role in biological processes, such as neuronal development, cardiovascular development, and immunological system Figure 1via the RhoA GTPase  , the hepAccording to the studies of West et al., NRP1 binds to FGF isoforms  and to their receptor FGFR-1, thus contributing to angiogenesis. The same team also showed that NRP1 binds to HGF, which is involved in various functions, such as cell proliferation, cell migration, and morphogenesis , 61. Thevia the regulation of TGF-β in endothelial cells , and lymphocytes, in particular regulatory T lymphocytes (LTreg) –121 and via the SEMA 3A/NRP1 signaling pathway  model of mice deficient in NRP1 on their T CD4+ lymphocytes (LT CD4+) and showed that mice presented aggravated EAE in contrast to mice over-expressing NRP1 on their LT CD4+, which did not develop EAE . This su pathway . We know pathway , 131. Al pathway . Further pathway . More re pathway . NRP2 is pathway , 132, an pathway . Further pathway .In addition to playing an immune-regulatory role, NRPs are also involved in the immune cell migration. For example, a study showed that SEMA 3A decreases the adhesion capacity of thymocytes expressing NRP1 and induces their migration by a repulsive effect . In the via MHC-I and MHC-II molecules, to present an antigen. APCs present antigen by forming an immunological synapse with LTs and thus stimulate their responses. NRP1 is expressed on both the surface of CDs and LTs and appears essential in their interaction to initiate the immune response , which have the ability, response . In partresponse . Moreoveresponse .Macrophages, another important player in immunity, also express NRPs , 126. ThThe immune system plays an essential role in our organism, especially during cancer when its immunological surveillance system is essential to control the tumor development and eliminate tumor cells. However, an important aspect of tumor progression is the ability of cancer cells to evade the monitoring and clearance by the immune system. Indeed, tumors cells avoid the immune system by different mechanisms, such as reducing the immune recognition, increasing the resistance to immune attacks, or creating an immunosuppressive microenvironment and recruiting specific immune cells that favor tumor growth and progression , 143. Bevia the SEMA-4A signaling pathway  cell responses , 135. Inr growth . Indeed,r growth .via its interaction with SEMA 3A, is involved in their migration. Indeed, the work of Casazza et al. shows that NRP1 controls the recruitment of TAMs within tumors. Its inhibition within macrophages causes a reduction of their pro-angiogenic and immunosuppressive effects and leads to an inhibition of tumor growth and metastasis (via the expression of IL10 or TGF-β for example (Another immune cell involved in anti-tumor immunity is the macrophage. Tumor-associated macrophages (TAMs) are strongly associated with tumorigenicity. NRP1, tastasis . A recentastasis . In this example .NRPs are, therefore, involved in anti-tumor immunity and it is questionable whether the same is true for brain tumors. The central nervous system was classically considered as an “immune-privileged site,” being immunologically inert and separated from the peripheral immune system by the blood-brain barrier (BBB). However, brain tumors represent a particular case in which the BBB is disrupted to various degrees, and therefore, acquired an innate immunity that may play a role in the development and growth of brain cancers. Most studies have focused on adult gliomas, and very few so far have focused on pediatric brain tumors. Several studies led by Miyauchi et al. have shown that NRP1 is expressed by macrophages and microglia associated with gliomas , 130. Inin vitro and in vivo have been developed  has been observed in pediatric tumors database. Moreover, NRP1 is involved in the activation of the immune cells, in particular in the interaction between T cells and DCs. NRP1 may also play a role in the regulation of immunogenicity of tumor cells, highlighted by the correlation between NRP1 and PDL-1. This role is not yet fully investigated, but we will focus our future research on this aspect. Numerous peptides or biological molecules that target NRP1 and whose efficacy has been tested eveloped  and moreeveloped . To concMD, PC, and CB contributed to redaction of this review. MD wrote the first draft of 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.