Carcinogenic aflatoxins can be inactivated by smectites  through adsorption and degradation. Proteins in gastric fluids can reduce smectite’s adsorption capacity for aflatoxins. The objective of this study was to evaluate the efficiency of smectites modified with organic nutrients in restricting the influence of proteins on aflatoxin adsorption. Arginine, histidine, choline, lysine, and vitamin B1 were selected to occupy part of the interlayer space of montmorillonite to achieve a smectite structure more selective for aflatoxin adsorption, but not for the large protein molecules. The unmodified montmorillonite had a maximum adsorption capacity of 0.2 mol/kg in the presence of pepsin. The vitamin B1-montmorillonite showed significant improvements in the aflatoxin affinity constant from 0.065 to 0.201  Aspergillus. Numerous adsorption experiments have demonstrated that smectites have a high adsorption capacity for aflatoxin. Montmorillonites can adsorb up to 20% (by mass) of aflatoxin. Animal experiments have shown that smectites can effectively reduce aflatoxin toxicity by adsorbing aflatoxins in the gastrointestinal tract when incorporated into an aflatoxin contaminated feed [Aflatoxins are carcinogenic secondary metabolites produced by fungi ted feed ,2,3.Variable temperature X-ray diffraction analysis demonstrated that aflatoxin molecules were adsorbed in the interlayer space of smectites and occupied the hydrophobic sites . It was sorption . The adsIn previous experiments, the efficiency of a bentonite  with a high aflatoxin adsorption capacity was tested in an in vivo experiment . The batDespite numerous successes in adsorption experiments that have demonstrated the high capacity of the clays in adsorbing aflatoxin in water, a complete animal recovery from aflatoxicosis in animal trials has not been observed after adding smectites to aflatoxin contaminated feed. Moreover, the quantity of incorporated smectite in the feed trials should adsorb several orders of magnitudes or more aflatoxins than those in the contaminated feed. The lower than expected performance of smectites in animal trials can be at least partially attributed to the complex compositions of the gastrointestinal fluids.Comparing the adsorption capacities of two smectites in water and in corn meal solution showed a significant AfB1 adsorption reduction in the corn meal solution . The lowA recent study indicated that the Ca-montmorillonite’s adsorption capacity (Qmax) for aflatoxin reduced from 0.52 mol/kg in water to 0.32 mol/kg in simulated gastric fluid . DivalenTo retain the high aflatoxin adsorption capacity of smectites in gastric fluids, the accessibility of proteins to the interlayer of smectite must be blocked by certain treatments, yet the treatments should keep the adsorption sites available for aflatoxin molecules.A common approach to modify the interlayer of smectites involves the intercalation of a cationic surfactant or small organic cations to increase the surface hydrophobicity of the clay. Jaynes et al.  intercalTo avoid the possible toxicity of the modifying organic or inorganic compounds, it would be more desirable to use non-toxic or low toxicity compounds such as food ingredients or organic nutrients to achieve an optimal interlayer space and surface polarity for selective adsorption of AfB1 . BarrienThe objective of this study was to evaluate the effectiveness of smectites’ modifications with five common organic nutrients in the aflatoxin B1 adsorption in simulated gastric fluid by restricting the interlayer access of proteins such as pepsin. Our goal was to let organic nutrients occupy part of the interlayer space of smectites to achieve a more desired interlayer space for aflatoxin adsorption, but not for the large protein molecules.The XRD diffraction patterns of the nutrient-montmorillonite (Mt) complexes showed a range of d(001)-values from 14.6 to 12.8 Å at room T . The intThe intercalation of the nutrient compounds in the montmorillonite was confirmed with the FTIR spectra of the nutrient-Mt complexes. The C=N vibration at 1671 cm complex . Interlad to C=O , was obsThe 1643 cmbrations , shifted choline . Both chWhen vitamin B1 was adsorbed by montmorillonite, the 1689 cmThe adsorption of pepsin resulted in different interlayer expansions in the montmorillonite . The basin Ca-Mt .When both AfB1 and pepsin were adsorbed, the Ca-Mt d001value was slightly lower than pepsin-Mt at room T . A similThe FTIR spectra of the Pep-nutrient-Mt complexes confirmed the presence of pepsin in the complexes as the major pepsin bands observed in the Ca-Mt spectrum were also present in the nutrient-MT complexes . The FTIIn the presence of pepsin, the nutrient-Mt complexes improved the aflatoxin adsorption as compared to Ca-Mt . The ArgThe FTIR spectra of the Aflatoxin-nutrient-Mt complexes showed the major bands of adsorbed aflatoxin molecules in the region from 1800 to 1200 cm−1 , similarMontmorillonites have been widely used as adsorbents for numerous organic compounds such as amino acids, vitamins, and various medical molecules. Early studies demonstrated the adsorption and interaction of amino acids by smectites. Jang et al.  investig et, al.  describe et, al. , when inThe thermogravimetric analysis of amino acids-montmorillonite complexes by Han et al.  showed aThe nutrient-Mt increased the amount of aflatoxin adsorbed in the interlayer in the presence of pepsin as a result of the nutrient-Mt complexes restricting the intercalation of pepsin. The efficiency varied possible due to the different nutrients’ loadings. Heating the Pep-nutrient-Mt complexes collapsed the interlayer to 13 Å in comparison to Pep-Ca-Mt, which maintained the interlayer at 15 Å. The significant difference in the d-values indicated that less pepsin was adsorbed in the nutrient-montmorillonite complexes than Ca-Mt.At room temperature, Chol-Mt had similar d-value as AfB-Pep-Chol-Mt. The lack of expansion indicated that choline in the interlayer favored the adsorption of aflatoxin on the hydrophobic sites, but in the pepsin solution alone, the interlayer expanded to about 15.5 Å.Among the five nutrient-Mt complexes, VB1-Mt was the most effective at improving the aflatoxin adsorption capacity and the affinity in the presence of the protein. The increased adsorption capacity from 0.35 mol/kg in Ca-Mt to 0.56 mol/kg in VB1-Mt in the presence of pepsin suggested that the nutrient-Mt potentially increased the adsorption of aflatoxin in animal experiments. Further in vivo trials are required to test the improved aflatoxin adsorption. The significant improvement of the Qmax and the affinity resembled previously observed isotherm values in simplified solution (water) . Ca-Mt sA complete pepsin restriction to the interlayer by the nutrient-Mt was not achieved in this study. The d001 basal spacing of the nutrient-Mt complexes collapsed to 10 Å after heating at 300 When modified with lysine, the basal space of montmorillonite showed only a slight expansion in the presence of aflatoxin and pepsin. This increase could be a change in the arrangement or conformation of the lysine or room humidity effect.VB1-Mt showed a higher d-value when both pepsin and aflatoxin were present in the solution. The high adsorption maxima, along with the reduced FTIR pepsin bands and the expanded basal spacing indicated a different aflatoxin molecule arrangement in the interlayer from the nearly flat monolayer observed in the AfB1-smectites formed in aqueous solutions. The adsorption isotherm was not L-shaped, but it resembled more an initial S-type shape.Deng et al.  demonstr748 cm−1 , which iThe band at 1235 cmIntercalating the five cationic and polar organic nutrients  into the interlayer space of montmorillonite improved the AfB1 adsorption in simulated gastric fluid by restricting the adsorption of pepsin. Yet, pepsin was not completely blocked. Among the five tested nutrients, vitamin B1 was the most effective at blocking the adsorption of pepsin and increasing the adsorption capacity and affinity of aflatoxin in the simulated gastric solution. AfB1 molecules could access the interlayer spaceof the nutrient-Mt complexes. The nutrient had small effects on the bonding mechanism of aflatoxin molecules to montmorillonite in the complexes.The <2 in water .Five commonly used nutrients in feed were selected as modifying organic anchors in the montmorillonite’s interlayer: three amino acids , one vitamin (thiamine), and a vitamin-like essential nutrient (choline). These small nutrient molecules are either cationic or can be protonated, and they are suitable to occupy the negatively charged sites in smectite without competing with the neutral non ionic aflatoxin molecules. Thiamin chloride (vitamin B1), L-lysine, and choline chloride were purchased from Sigma Aldrich . The L-arginine monohydrochloride and L-histidine monohydrochloride monohydrates were purchased from VWR. A stock solution of 1000 mg/mL was prepared in DI water for each organic compound, and the pH of each stock solution was recorded.Pepsin (from porcine gastric mucosa) was obtained from Sigma Aldrich. The pepsin solution was prepared by dispersing 624 mg of pepsin in 500 mL of DI water . The pepCa-montmorillonite (Ca-Mt) dispersions, containing 10 mg of the clay mineral, were mixed with 10 mL of the 1000 mg/mL stock solution of each organic nutrient. The samples were shaken overnight on an orbital shaker. The nutrient-montmorillonite (nutrient-Mt) complexes’ dispersions were centrifuged at 4000 rpm for 40 min and washed twice with DI water. After removing the supernatant, 5 mL of DI water were added to each sample to obtain a 2 mg/mL clay dispersion .To examine if the nutrients intercalated in the montmorillonite, the basal spaces d(001) of the nutrient-montmorillonite complexes were monitored by XRD at elevated temperatures. The nutrient-Mt complex dispersions were air died on silicon wafers and placed in an Anton Paar reactor chamber XRK900 on a Bruker D8 advance diffractometer. The initial temperature in the chamber was set to 30 The nutrient-Mt complexes were loaded with AfB1, pepsin, and AfB1 plus pepsin to check the interlayer accessibility. To prepare the AfB1-nutrient-Mt complexes, an aliquot of 0.5 mL of the nutrient-Mt dispersion, containing 1 mg of the complex, was mixed with 20 mL of an 8 ppm AfBAll nutrient-montmorillonite complexes loaded with AfB1, pepsin, or AfB1+pepsin were air dried on zero background silicon slides for X-ray diffraction analysis (XRD) at room temperature. Then, the slides were heated at 300 To investigate the interactions among the aflatoxin, nutrients, and montmorillonite, the montmorillonite complexes loaded with nutrients, aflatoxin, and the nutrient solutions were air dried on ZnS windows, and FTIR spectra were recorded in transmission mode in a Spectrum 100 Perkin Elmer FTIR spectrometer with 32 scans and a resolution of 2 cmAflatoxin adsorption isotherms in the pepsin solution of the nutrient-Mt complexes were used to evaluate their capacity in blocking proteins from the interlayer. The isotherms were prepared following the procedure described in Barrientos et al. . Fifty m


We assessed adherence to government recommendations implemented shortly after the introduction of COVID-19 in Mozambique in March 2020, through two online cross-sectional surveys in April and June 2020. We quantified adherence to preventive measures by a composite score comprising of five measures: physical distancing, face mask use, hand hygiene, cough hygiene, and avoidance of touching the face. 3770 and 1115 persons participated in the first and second round respectively. Wearing face masks, regular handwashing and cough hygiene all reached compliance rates of over 90% while physical distancing and avoiding to touch the face reached a compliance rate of 80–90%. A multivariable model investigating factors associated with adherence found that being older, more educated, and belonging to the healthcare sector increased the odds for higher adherence. Private workers and retired people, respondents receiving COVID-19 information through social media, and those who reported flu-like symptoms were less likely to adhere. 6% of respondents reported flu-like symptoms which aligned with the WHO clinical definition of COVID-19, suggesting low level community transmission. In conclusion, most respondents in this online survey in Mozambique complied well with strategies to prevent COVID-19. Whether the good preventive behaviour explains the low grade COVID-19 transmission requires further study. On 11 March 2020 COVID-19 was declared a global pandemic by the World Health Organization  , DeDuring both surveys a relatively large number of respondents (18%) reported experiencing at least one flu-like symptoms during the two weeks preceding the survey. In about 6% of all respondents, the flu symptoms met the criteria of the WHO clinical definition of suspected COVID-19 infection. However, all 69 participants (54 without a recent flu–like illness) with available COVID-19 results tested negative. This suggest low grade COVID-19 transmission in Mozambique during the period of the surveys. This has been confirmed by several provincial sero-surveys in Mozambique with community sero-prevalences ranging from less than 1%  to 5% 1.Persons with low adherence were more likely to report a flu-like illness. This was contrary to our expectations; we anticipated that during a COVID-19 outbreak, the experience of flu-like symptoms would cause people to be more careful with their health and consequently improve their adherence to COVID-19 preventive measures. However, a more likely interpretation of this finding is that the flu-like symptoms were the consequences of low adherence to preventive measures.Research has shown that face masks are an important tool to prevent the spread of respiratory pathogens . HoweverA higher level of education, with the exception of postgraduate education level (only 7 respondents), was associated with better preventive behaviours. Participants who received COVID-19 information from social media were less likely to be adherent with the preventive measures suggesting that social media may have had a negative effect on the preventive behaviour of people. Therefore, it is important that governments disseminate clear, evidence based messages to the public about COVID-19 preventive measures. Being older was also related to high adherence to preventive measures. This is an important finding as older age is a risk factor for severe COVID-19 disease, and therefore the protection of the elderly is a priority during the current outbreak.Despite the fact that most COVID-19 infection are reported from Maputo City, adherence with the COVID-19 preventive measures was relatively high in this province. The explanation for this may be that in this province which was the epicenter of the local outbreak, people considered themselves more at risk for COVID-19 infection and therefore may be more motivated to adhere to the preventive measures.Some limitations of the study should be mentioned. To keep the questionnaire short and easier to fill, we assessed adherence to preventive measures using yes/no questions. Using Likert-like scale questions would have provided more precise information, albeit making the questionnaire lengthier. Similarly, the use of the term “regular” in quantifying the frequency of handwashing was not very precise and could be interpreted differently by different respondents. Moreover, self-reported responses may not reflect the real-life behaviour of participants.People with no or limited internet access  were not able to participate in the research. People without internet connection generally will be older and are less educated but also may have been less exposed about fake news circulating on the internet. Therefore, our respondents cannot be considered as representative of the general population in Mozambique. As this was a cross-sectional survey, causality is difficult to assess, as we pointed out above when assessing the relationship between adhering to preventive measures and having flu-like symptoms.In conclusion, these online surveys in Mozambique showed that most respondents complied well with most strategies to prevent COVID-19 transmission. In particular, adherence to face mask use and regular hand washing was high (more than 90%). However, a considerable number of participants attended markets where physical distancing can be challenging, which can be a risk factor for contracting COVID-19. Based on reported symptoms and data from local sero-surveys, there seems to have been low grade community COVID-19 transmission during the period of the surveys. Whether good COVID-19 preventive behaviour in Mozambique is responsible for this low grade of COVID-19 transmission requires further study. Complementing our study findings with qualitative research particularly among populations at higher risk for COVID-19 infection would certainly provide a better picture of the COVID-19 behavioural landscape in the country.


X-symmetry points of the Brillouin zone, we do not find these gaps to exhibit band inversion so that HoSb does not host a Dirac semimetal state. Our magnetotransport measurements indicate that HoSb can be characterized as a correlated nearly-complete electron-hole-compensated semimetal. Our analysis reveals that the nearly perfect electron-hole compensation could drive the appearance of non-saturating XMR effect in HoSb.The rare-earth monopnictide family is attracting an intense current interest driven by its unusual extreme magnetoresistance (XMR) property and the potential presence of topologically non-trivial surface states. The experimental observation of non-trivial surface states in this family of materials are not ubiquitous. Here, using high-resolution angle-resolved photoemission spectroscopy, magnetotransport, and parallel first-principles modeling, we examine the nature of electronic states in HoSb. Although we find the presence of bulk band gaps at the  The discovery of TIs has spurred the exploration of gapless nontrivial states beyond the insulators to include a great variety of topological semimetals such as the Dirac-, Weyl-, and nodal-line/loop semimetals in which the bulk bandgap opening is prohibited by crystalline symmetries9. These developments have opened up exciting new routes for finding exotic quantum phases and improved materials platforms for constructing low-power electronics/spintronics devices10. A distinct nontrivial 12 where the TSSs overlap with the bulk states. The rare-earth monopnictide (REM) family is drawing special interests as a rich playground for investigating Dirac fermionic excitations. REMs have been predicted to host topological Dirac semimetal as well as 3D TI states13 and exhibit extremely large magnetoresistance (XMR)20. Although a good deal of work in the literature concerns the linearly-dispersing states and their role in driving XMR in the REMs, a robust conclusion in this regard remains a matter of debate. Although the conventional explanation for the XMR effects involves electron-hole compensation22, the observation of XMR in the REM family and numerous other topological materials such as 23, 24, 25, and TaAs26 has ignited the possibility that XMR might have its origin in the forbidden backscattering channels of topological materials26. Notably, TSSs have been reported in LaBi, LaSb, CeSb, and CeBi by various groups36 whereas LaAs, LuBi, YBi, YSb, and CeSb have been suggested to be topologically trivial materials37. The presence of a Dirac semimetal state is suggested in NdSb38 and DySb18. With this background in mind, further experimental and theoretical studies are needed to gain an understanding of the nature of Dirac-like states and how they are connected with XMR in the REM family.Topological insulators (TIs) with novel properties such as extreme magnetoresistance and high carrier mobility are currently attracting intense interests in condensed matter and material science communities. A TI supports the presence of gapless topological surface states (TSSs) with Dirac-cone-like energy dispersions within an inverted gap between the bulk conduction and valence bandsf-electrons provide strongly-correlated, tunable magnetic ground states in the REM family. Moving from La to Lu in the lanthanide series, one observes a nonmagnetic to ferromagnetic transition. HoSb that supports an antiferromagnetic (AFM) ground state39 is not a well-studied member of the REM family. It displays a magnetic transition from an MnO-type AFM arrangement to a HoP-type ferromagnetic arrangement under external magnetic field40. More recently, an unusual XMR has been reported in HoSb42. Here, we report the observation of a gapped state at the X point of the BZ in HoSb. Using angle-resolved photoemission spectroscopy (ARPES) along with first-principles calculations and magneto-transport measurements, we examine in-depth the electronic structures of this material. Our analysis reveals the presence of a highly anisotropic Dirac-like cone at the X point. Our experimental data and theoretical results identify a small gap around 470 meV below the Fermi level. More importantly, our calculations of the Rare-earth elements with their Fm-3m like other members of the REM family  Å. Here, yellow and blue balls correspond to the Ho and Sb atoms, respectively  effects and treating mentclass2pt{minimX point, we clearly see two concentric elliptical-shaped pockets. However, moving to higher binding energies, the elliptical-pockets evolve into point-like features around 460 meV, which indicates the electron-like nature of the bands around the X point. Importantly, our calculations show the presence of a local band gap near this binding energy at the X point. Around the binding energy of 600 meV, size of the elliptical pocket increases, suggesting that the possible bulk bandgap lies around this binding energy. We will return below to present additional photon energy-dependent dispersion maps around the X points in order to ascertain the nature of the bands from our experimental measurements. Figure We now discuss the bulk band structure and the Fermi surface using our measured constant energy contours for the (001) surface, see Fig. entclass1pt{minimaX point is about 90 meV. There is a continuous band gap between the valence and conduction bands so that the 44, we find a trivial topological invariant Figure entclass1pt{minimai) was applied along the i and 41. Application of magnetic field strongly increases electrical resistivity and 41. The field dependent resistivity saturation at low temperatures has not been observed in the non-magnetic members of the REM family28. The origin of such a plateau has been recently attributed to the presence of a nearly perfect electron-hole carrier compensation and the high mobility of the compensated semimetals45. Figure R(B) and R(0) are resistances with and without the magnetic field, respectively. As seen from the inset of Fig. B) curves mark a metamagnetic transition that is related to the change in the magnetic structure with the increase of field46. At 41. In this connection, we measured MR in pulsed magnetic fields up to 30 T  curve at 4 K can be described by the form MR 45. In addition, for 49.Our ARPES measurements indicate that HoSb is a trivial semimetal. In order to study the transport properties of HoSb, we used a micron-sized sample prepared by plasma focused ion beam (PFIB) microscope. Magneto-transport measurements were performed on a FIB50, HoSb offers a unique platform for exploring the interplay between XMR, magnetism and topology in an antiferromagnetic matrix.We have carried out in-depth photoemission and transport measurements on HoSb single crystals along with parallel first-principles modeling of the electronic structure of this member of the REM family. Our ARPES measurements show the presence of Dirac-like cone with a small gap at around 470 meV below the chemical potential, but our analysis shows that HoSb is a trivial and not a topological Dirac semimetal. The observed experimental bulk band gap at X point is around 120 meV which is consistent with the theoretically predicted value. Magnetoresistance is found to be large and non-saturating, even at a magnetic field as large as 30 T. The characteristic behavior of electrical resistivity at low temperatures indicates that HoSb is likely a new electron-hole-compensated semimetal. Notably, the XMR effect has also been reported in other nontrivial members of the REM family suggesting that the presence of non-trivial state might not be directly responsible for the XMR. However, the high carrier mobility associated with the topological states might play a role in generating the XMR. Furthermore, our magnetoresitivity measurements up to 30 T show a similar unsaturated behavior below and above 43. The crystal structure was determined by X-ray diffraction on a Kuma-Diffraction KM4 four-circle diffractometer equipped with a CCD camera using Mo KSingle crystals of HoSb were grown by the Sn flux technique as described elsewhereSynchrotron-based ARPES measurements of the electronic structure were performed at the Advanced light Source (ALS) beamline 4.0.3 with a Scienta R8000 hemispherical electron analyzer. The samples were cleaved in situ in a ultra high vacuum conditions  method. The electrical resistivity and magnetoresistivity were measured using a Quantum Design Dynacool-9 device in magnetic field up to 9 T. Magnetoresistance measurements at pulsed magnetic fields up to 30 T were performed at NHMFL, and LANL using the same HoSb sample as that used in DC fields. For both measurements, a standard four probe measurement technique was applied.51 as implemented in the VASP suite of codes53. The exchange-correlation functional was treated using SCAN meta-GGA54. An energy cutoff of 400 eV was used for the plane-wave basis set and a 55. The surface energy spectrum was obtained by using the iterative Green’s function method via the WannierTools package58.Electronic structure calculations were performed within the framework of the density-functional theory (DFT) using the projector-augmented-wave (PAW) methodSupplementary information.


Peripheral artery disease is caused by atherosclerosis of lower extremity arteries leading to the loss of blood perfusion and subsequent critical ischemia. The presence of diabetes mellitus is an important risk factor that greatly increases the incidence, the progression and the severity of the disease. In addition to accelerated disease progression, diabetic patients are also more susceptible to develop serious impairment of their walking abilities through an increased risk of lower limb amputation. Hyperglycemia is known to alter the physiological development of collateral arteries in response to ischemia. Deregulation in the production of several critical pro-angiogenic factors has been reported in diabetes along with vascular cell unresponsiveness in initiating angiogenic processes. Among the multiple molecular mechanisms involved in the angiogenic response, protein tyrosine phosphatases are potent regulators by dephosphorylating pro-angiogenic tyrosine kinase receptors. However, evidence has indicated that diabetes-induced deregulation of phosphatases contributes to the progression of several micro and macrovascular complications. This review provides an overview of growth factor alterations in the context of diabetes and peripheral artery disease, as well as a description of the role of phosphatases in the regulation of angiogenic pathways followed by an analysis of the effects of hyperglycemia on the modulation of protein tyrosine phosphatase expression and activity. Knowledge of the role of phosphatases in diabetic peripheral artery disease will help the development of future therapeutics to locally regulate phosphatases and improve angiogenesis. The bibliographic search was performed in the PubMed and Google Scholar databases. All papers published before June 2020 were included in the review. A large set of keywords was used including growth factor, tyrosine kinase receptor, diabetic PAD, angiogenesis, blood flow reperfusion, endothelial dysfunction, endothelial cells, perivascular cells, hindlimb ischemia, phosphatase, and hyperglycemia.Current data reveals that more than 400 million people are suffering from diabetes mellitus (DM) and projections estimate that numbers of this world-wide issue will almost triple by 2030 . DM is ain vitro and in vivo) will be reported to highlight the underlying molecular mechanisms leading to the alteration of the angiogenic process following hindlimb ischemia in diabetes. Findings based on other diabetic macrovascular complications sharing a similar pathophysiology as PAD will be reported where preclinical data was not available in the context of diabetic PAD.Angiogenesis is a complex, dynamic, and well-coordinated physiological process leading to the formation of new vessels from pre-existing vasculature in response to ischemia. This process involves temporal interactions between multiple growth factors and vascular cells, which have been extensively summarized in excellent reviews , 12. BriFrom the extensive list of pro-angiogenic factors, VEGF is by far the most clinically investigated. As a result of blunted cellular response to hypoxia, hypoxia-inducible factor-1 alpha (HIF-1α) itself and most of its target genes, including VEGF, are reduced in a diabetic state . Thus, aUnlike VEGF, the impact of hyperglycemia on PDGF expression and activation in the context of PAD has been poorly clinically investigated. To date, only one pilot study has reported lower serum levels of PDGF-BB in T2D patients with a previous cardiovascular event compared to those that have not  Table 1Table 1. Two consecutive cross-sectional studies investigated the serum levels of Ang-1 and Ang-2 in control and T2D patients, with or without CVD, including PAD , 24 Tab. Only Anet al. reported lower numbers of CXCR4-positive circulating cells, SDF-1's main receptor in T2D patients which was further decreased in T2D individuals suffering from peripheral atherosclerosis  that mimic the pathophysiology of PAD and allow to assess ischemia-induced angiogenesis/arteriogenesis as well as subsequent blood flow reperfusion. Several different surgical approaches are used to induce ischemia in the lower limb of small animals , 34. Thedb/db T2D mice, which was illustrated by a significant reduction in capillary density and blood flow reperfusion 35 days post HLI-surgery  1/2 in HUVEC simultaneously exposed to hyperglycemia and CoCl2-induced hypoxia compared to controls leading to reduced migration  activity will trigger abnormalities in cellular processes including proliferation, migration, and differentiation, thereby resulting in a global deregulation of angiogenesis . AdditioPTPN1) is an intracellular ubiquitous PTP. The systemic deletion of PTP1B results in viable and fertile mice with enhanced insulin sensitivity and accelerated wound healing  is a cytoplasmic tyrosine phosphatase. SHP-1 null mice showed chronic systemic inflammation and died within 3 weeks of birth from interstitial pneumonia  is a ubiquitously expressed non-transmembrane phosphatase sharing a similar structure and phosphatase activity mechanism as SHP-1. Unlike SHP-1, the global deletion of SHP-2 is embryonically lethal  belongs to the receptor PTP (PTPR) family anchored at the cell membrane. VE-PTP is exclusively expressed in EC  is the second member of the PTPR family and shares mechanistic analogies with VE-PTP. Nevertheless, since DEP-1 expression is not restricted to EC, it can bind a larger range of pro-angiogenic RTK mainly VEGFR2  is an intracellular ubiquitously expressed non-receptor PTP. TC-PTP was found to be expressed in both EC and VSMC  is a ubiquitous lipid and protein phosphatase. PTEN is also the main regulator of the PI3K/Akt pathway by dephosphorylating phosphatidylinositol-3,4,5-trisphosphate (PIP3) into phosphatidylinositol-3,4-bisphosphate (PIP2), therefore indirectly preventing RTK-induced Akt activation  are either cytoplasmic or nuclear phosphatases acting upon tyrosine or serine/threonine residues which are responsible for dephosphorylation and regulation of MAPKs . Therefore, DUSPs indirectly participate in the regulation of RTK-induced angiogenesis. Several studies reported a pro-angiogenetic role of multiple DUSPs. For instance, silencing DUSP1 blunted VEGF-A-induced migration , 96, proprouting . Conversprouting . High glprouting . Recentlprouting .In the HLI model, the deletion of DUSP1 led to a significant reduction in blood flow reperfusion at days 3 and 7 post-surgery, gradually recovering to levels similar to controls by day 28. Muscle vessel density was altered in the same time frame highlighting the critical role of DUSP1 in the early response to hindlimb ischemia . KnockinHyperglycemia is deeply involved in the progression of macrovascular complications by deregulating cellular responses to hypoxia. Diabetic PAD is characterized by insufficient angiogenesis of the lower extremities which is attributed to impaired production of pro-angiogenic factors and defects in signal transduction in both vascular and perivascular cells. Although diabetes-induced alteration of VEGF signaling is extensively proven, there is still a lack of conclusive large-scale clinical studies concerning other growth factors such as PDGF and angiopoietins. Hence, larger clinical studies supported by suitable preclinical mechanistic models are required to guide and optimize the development of new therapeutic targets based on the administration of exogenous growth factors which are currently lacking sustainable effects. The source of vascular complications in diabetes is tremendously multifactorial and PTP have progressively emerged as potent players in vascular cell unresponsiveness to growth factors. Although the role of PTP in the modulation of pro-angiogenic RTK signaling is well-documented at a physiological level, their regulation in the complex hypoxic and hyperglycemic environment in PAD remains partially understood. Because the expression of PTP is not strictly limited to the endothelial lineage but extended to perivascular cells, the use of simultaneous cell and phosphatase-specific knockout models represent the next step in improving knowledge on this subject. In addition, since diabetic PAD is in part characterized by vascular and perivascular cell unresponsiveness to pro-angiogenic growth factors due to the upregulation or downregulation of PTP activity, the development of PTP inhibitors and activators is also an exciting avenue to modulate pro-angiogenic RTK activity . From a The original contributions presented in the study are included in the article/supplementary materials, further inquiries can be directed to the corresponding author/s.CM and PG conceived the idea for the review. CM performed the literature search and wrote the manuscript. PG and MR critically revised the work. 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.


Autophagy is a lysosomal degradation pathway for intracellular components and is highly conserved across eukaryotes. This process is a key player in innate immunity and its activation has anti-microbial effects by directly targeting pathogens and also by regulating innate immune responses. Autophagy dysfunction is often associated with inflammatory diseases. Many studies have shown that it can also play a role in the control of innate immunity by preventing exacerbated inflammation and its harmful effects toward the host. The arms race between hosts and pathogens has led some viruses to evolve strategies that enable them to benefit from autophagy, either by directly hijacking the autophagy pathway for their life cycle, or by using its regulatory functions in innate immunity. The control of viral replication and spread involves the production of anti-viral cytokines. Controlling the signals that lead to production of these cytokines is a perfect way for viruses to escape from innate immune responses and establish successful infection. Published reports related to this last viral strategy have extensively grown in recent years. In this review we describe several links between autophagy and regulation of innate immune responses and we provide an overview of how viruses exploit these links for their own benefit. Cellular catabolism is ensured by both the Ubiquitin Proteasome System and the process of autophagy acting in a coordinated manner in order to maintain homeostasis . HoweverUpon viral infection, host cells possess an arsenal of innate responses to protect themselves and their neighbors, counteracting pathogen replication and spread. Pattern Recognition Receptors (PRRs) orchestrate the detection of Pathogen-Associated Molecular Patterns (PAMPs), triggering an anti-microbial response by an important shift in transcriptional activity . Upon stper se, but also to autophagy-independent functions of autophagy-related proteins. Thus, even if autophagy is not the only regulatory pathway involved, it is now considered an important regulator of the innate immune response. In this review, we describe the different innate immune signaling pathways that can be counteracted or used by viruses for their own benefit.Autophagy is involved in major biological processes, for instance in development , cellulaThe term autophagy refers to “self-eating” and is the lysosomal degradation of intracellular components. It includes three distinct mechanisms: microautophagy, Chaperone Mediated-Autophagy (CMA), and macroautophagy .Microautophagy is a process involving the protrusion or invagination of lysosomal membranes, leading to engulfment of cytosolic components directly into the lysosome for their subsequent degradation  10). CM. CM10). Historically deciphered in yeast models, autophagy involves more than 40 AuTophaGy-related (ATG) genes. Eighteen of them are highly conserved among eukaryotes and comprise the core autophagy machinery –16. Brie(i) The initiation step: This is dependent on two main complexes: the uncoordinated-51-like protein kinase (ULK) complex. This is composed of ULK1 or ULK2, ATG13, Focal adhesion kinase family Interacting Protein 200 (FIP200), and ATG101 . The sec(ii) The elongation step: This corresponds to phagophore expansion, in which two sophisticated ubiquitination (Ub)-like conjugation complexes are involved. The first one is responsible for the conjugation between ATG5 and ATG12, mediated by the E1-activating enzyme ATG7 and the E2-conjugating enzyme ATG10. Then, ATG16L1 binds to ATG5-ATG12 conjugates in a non-covalent manner . The sec(iii) The maturation step: This consists in the fusion between the autophagosome and the lysosomal compartment. This step is achieved by the UV Resistance-Associated Gene (UVRAG)-containing class III PtdIns3K complex. Many other actors are also involved, such as the Rab7 GTPase and the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex. This is composed of Syntaxin 17 (STX17), SyNaptosomal-Associated Protein 29 (SNAP29), and Vesicle-Associated Membrane Protein 8 (VAMP8) .Fusion leads to the formation of a structure called the autolysosome, in which the degradation of the sequestered materials occurs by lysosomal hydrolases. It is noteworthy that the ATG8-PE conjugate is present on autophagosomes throughout the entire process of autophagy and is degraded in the lysosome, making it a good marker for monitoring autophagic flux. Finally, resulting metabolites are transported into the cytosol in a recycling step .Although initially considered a random mechanism, it is now well-established that autophagy can be highly specific through the action of selective autophagy receptors (SARs) . These pSARs are able to bind substrates and lead them to the expanding phagophore. On one hand, substrates are often ubiquitinated and therefore recognized by the ubiquitin-binding domain present on most SARs. On the other hand, SARs contain an LC3-Interacting Region which allows the targeting of selected substrates to phagophores . Many SAATG8 proteins, hereafter referred to as LC3 , are present on autophagic membranes from the elongation to the degradation step in their conjugated form (LC3-PE known as LC3-II). Therefore, monitoring LC3-II protein expression is a classic approach in studying autophagic flux . HoweverAutophagy is an intrinsic pathway of innate immunity. Consequently, autophagy intersects with all the innate immune signaling pathways activated upon viral infection. Indeed, it favors the innate immune response by participating in cytokine secretion and by down-regulating the immune response to prevent deleterious effects of prolonged immune activation. TLRs are classified in two subfamilies based on their localization : those eDrosophila, acts as a selective autophagy receptor for IKK-β in order to negatively regulate NF-κB responses and prevent constitutive inflammation. However, although NEMO in Drosophila contains a LC3-Interacting Region, it is not the case for its mammalian orthologs and they are therefore not involved in IKK-β-selective degradation, showing an evolutionary divergence in NEMO function   and Humafunction . In addifunction , while tfunction . Finallyfunction .Nucleotide-binding oligomerization domain, Leucine-rich Repeat and Pyrin domain-containing (NLRP) receptors are PRRs belonging to the NOD-Like Receptor (NLRs) family. Their stimulation with a specific PAMP leads to the formation of a structure named the “inflammasome.” Upon activation, NLRPs recruit Apoptosis-associated Speck-like protein-containing a CARD (ASC) proteins and mediate their binding with Caspase-1 (CASP1). In turn, the inflammasome triggers the catalytic activity of CASP1, which is able to cleave pro-interleukins, such as pro-IL-1β and pro-IL-18, leading to their release . The infSeveral NLRs have the ability to form inflammasomes, such as NLRP1, NLRP3, and NLR family CARD domain-containing protein 4 (NLRC4). Some additional sensors have the same ability, including the Absent In Melanoma 2 (AIM2) protein or pyrin . During Atg5 genetic variants and Crohn's disease is associated with Atg16l1 and Irgm1 genetic variants. First evidence of a role for autophagy in inflammasome activation was described in Dextran Sulfate Sodium (DSS)-treated mice and LPS-treated macrophages both depleted of ATG16L1 expression. The absence of ATG16L1 led to an increase in inflammasome activation, higher levels of IL1β, and enhanced acute colitis in mice  and Melanoma Differentiation-Associated 5 (MDA-5) are the two mains sensors involved in this recognition. They are structurally composed of a DExD/H-box helicase and a C-terminal domain, both important for RNA recognition . RIG-I rThe first report demonstrating crosstalk between components of autophagy and the RIG-I/MAVS signaling pathway involved the ATG5-ATG12 complex independently of autophagy. This complex is able to prevent RIG-I/MAVS association by binding to their CARD domains, resulting in the inhibition of their activation . BECN1 iTo conclude, a recent study has shown that the RIG-I pathway induction is able to induce autophagy in several cell types. Indeed, upon RIG-I activation, BECN1 interacts with TRAF6 located on mitochondria and undergoes a K63-polyubiquitination that triggers autophagy. This process could reveal a negative feedback regulation of RIG-I/MAVS activation , howeverDNA is normally found in the nucleus or mitochondria of eukaryotes. Therefore, the presence of DNA in the cytosol manifests either microbial infection or cell damage. One of the main cytosolic DNA detection pathways depends on the cyclic GMP–AMP Synthase (cGAS) sensor. DNA binding of cGAS triggers conformational changes and activates its enzymatic activity . cGAS caActivation of the cGAS-STING signaling pathway leads to relocation of STING from the ER to the ERGIC and Golgi apparatus. Components of autophagy are crucial to prevent its aberrant activation. Indeed, STING is found on single-membrane ATG9 and LC3B-positives vesicles after double-stranded DNA treatment and this location is essential for the regulation of its activity . BesidesDnase II-depleted cells is directed to lysosomal degradation preventing STING activation  binding protein 1 (G3BP) degradation by autophagy. A recent study established that picornaviruses dampen type I IFN production  by inducMany studies on STING regulation by autophagy have been carried out in the HSV-1 infection model. Correspondingly, in infected macrophages STING is targeted to p62/SQSTM1-mediated autophagic degradation, a process responsible for decreasing the levels of pro-inflammatory cytokines . This dePLP2-TM viral proteins from several betacoronaviruses, such as SARS COV-1 and MERS, induce the interaction of STING with BECN1. This complex negatively regulates STING activity and therefore type I IFN production . AdditioOther components of innate immune responses are also targeted by autophagy during viral infections. For example, RUBICON is able to bind to IRF3, a process that inhibits its activation and favors the replication of several viruses, such as VSV and the H1N1 strain of IAV . During A constant selective pressure is imposed on viruses by their hosts, leading pathogens to adapt in order to replicate efficiently. Consequently, even if autophagy is known to be an important process in fighting viruses, some viruses have evolved various strategies to exploit it for their own benefit. In recent years, an increasing number of studies have reported a role of autophagy in the regulation of the immune system and how this can be hijacked by pathogens. Interestingly, this hijacking seems to be a common feature between pathogenic viruses independently of their genomic structures or their specific hosts, likely because autophagy is occurring in all eukaryotic cells. We describe in this review the current knowledge still expanding in the field. The description of the regulation of innate immunity by autophagy is often focused on one single innate immune pathway. However, several reports indicate a cross-talk between multiple cellular immune responses , showingin vivo. Many studies remain descriptive and further work is required to decipher the related mechanisms. It would be particularly important to characterize the effect of autophagy-modulating drugs during viral infections. Moreover, the recently discovered LAP and LANDO processes suggest that there is still a great deal to discover with respect to co-evolution between viruses and their hosts.It is well-known that innate responses can be deleterious by inducing an uncontrolled inflammation in many viral infections, leading to severe symptoms and sometimes death. This is particularly well-illustrated in the case of the SARS-CoV2 infection, responsible for the COVID-19 pandemic. In this context, autophagy could be considered as an important process regulating both viral replication and the innate immune response. Only a few studies have been conducted to study the link between the control of innate responses by autophagy and the outcome of the infection Humanity is facing the emergence of a large number of new pathogens, including zoonoses, particularly due to climate change and human activity. In this light, we will undoubtedly face many new viral emergences in the future. Therefore, the study of infectious diseases is essential to anticipate these future challenges.Overall, this review shows that autophagy plays a central role in host/pathogen interactions, in particular in the immune response and its hijacking by viruses. Thus, it seems essential to develop our knowledge in this field in order to be able to uncover new therapeutic strategies that combine effects on both viral replication and on the host immune response to infections.BP, VR-H, and LE have all contributed to the writing 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.


Increasing studies indicated that microRNA-203 (miR-203) may play an important part in the prognosis of CRC. Nevertheless, the prognostic and influential mechanism of miR-203 expression in CRC remains to be inconclusive. Accordingly, we conducted the current study to investigate the biomarker performance of miR-203 in CRC.In the present study, we conducted an evidence synthesis of the published literatures to identify the prognostic roles of miR-203 in patients with CRC. Moreover, several bioinformatics methods were applied for exploring the biomarker roles of miR-203.P = 0.021) for CRC. The gene Ontology (GO) analysis indicated that miR-203 targets were primarily involved in a series of GO items closely associated with the molecular pathogenesis of CRC. The pathway analysis exhibited the potential signal pathways of miR-203 involved in CRC including pathways in cancer, wnt pathway, prolactin signaling pathway, proteoglycans in cancer, FoxO pathway, focal adhesion and Ras pathway. By constructing a protein-protein interaction (PPI) network of the targets of miR-203, ten crucial proteins and a significant network module were retrieved and found to serve important roles in the molecular pathogenesis of CRC.It was demonstrated that elevated miR-203 expression was clearly related to worse overall survival (HR: 1.55, 95% CI: 1.07–2.24, Our results indicated that miR-203 may function as a promising biomarker to monitor CRC survival outcomes and progression. Notably, large-scale prospective cohort studies and biological experiments are required to confirm our conclusions. Colorectal cancer (CRC) is a significant health burden in both men and women, responsible for the third leading factor of cancer-related death . In spitMicroRNAs have been recognized as a subset of small and single-stranded noncoding RNAs which have emerged to be key regulators in various cellular processes containing differentiation, proliferation, apoptosis and stress response . In humaAmong these microRNAs, microRNA-203 (miR-203) is one of most commonly dramatically studied microRNAs in human cancers, with the location of the chromosome 14q32.33 . PreviouThus, we performed a comprehensive study of all eligible literatures to identify the relevance between miR-203 expression status and clinical outcomes of CRC patients. Moreover, an integrated bioinformatics analysis was also conducted for further understanding of prognostic value of miR-203 and its potential biomarker mechanism in CRC.The first evidence synthesis part was implemented following strictly with the guidelines of the PRISMA statement . RelevanInclusion criteria were as follows: 1) investigated the association between miR-203 status and CRC overall survival (OS); 2) diagnosed CRC with surgical pathology; 3) provided the hazard ratio (HR) and 95% confidence intervals (CIs) directly or key information for estimating these parameters indirectly through Kaplan–Meier curves and original survival data.Studies were excluded based on these exclusion criteria a: 1) lacked sufficient data; 2) letters, reviews, case reports or conference abstracts; 3) non-English studies.All included studies were independently assessed and extracted the data by two investigators (QP and YS). The following details of each article were recorded: i) first author’s last name and time of publication; ii) study location, patients number, distribution of age and gender and tumor stage; iii) miR-203 assay specimen and assessment method; iv) HRs and 95%CIs of elevated expression levels of miR-203 for OS. If the studies provided multiple types of survival data, we preferred the results of multivariate analysis as it considered confounding factors and has been demonstrate to be more accurate. If HRs were not offered by the original article, the statistical variables may be estimated from the Kaplan-Meier curves following the previously described method .We applied Newcastle-Ottawa scale for evaluating the quality of each eligible article which was judged on three levels: selection, comparability, and outcome . High qu2 statistic to evaluate the heterogeneity of the studies [P < 0.05), random-effects model would be taken to assess the combined effect size. If not, fixed-effects model may be selected [P-value < 0.05 would be deemed as statistically significant.For the data pooling, HRs and the 95% CIs were calculated to elucidate the relevance between the expressing level of miR-203 and clinical survival. We selected Cochran’s Q test and Higgin’s I studies . When obselected . We usedselected . In ordeselected . P-valueThe target genes of miR-203 were predicted using miRTarBase which is a database covers experimentally proved microRNA-target regulatory pairs based on reporter assays, western blot, or microarray experiments with up-regulated or down-regulated of microRNAs . UpdatedP < 0.05.For determining the functions of miR-203, we implemented a comprehensive enrichment analysis with the targets of miR-203. Gene Ontology (GO) has been recognized as a commonly selected approach in the annotation of a large number of proteins while the KEGG pathway analysis could provide more valuable functional data about how molecules or genes are networked in different biological pathways , 20. In P-value < 0 .05 would be reflected as statistically significant.To gain the in-depth understanding of the biological functions of miR-203, we employed a PPI network analysis with the target genes of miR-203. We applied the Search Tool for the Retrieval of Interacting Genes (STRING) database for retrieving the predicted interactions among the regulated proteins of miR-203 . A combiThe workflow of research identification process was illustrated at Fig. n = 6) or serum (n = 5). The expression of miR-203 was measured by quantitative real-time polymerase chain reaction (qRT-PCR). All the studies had scores from 6 to 9, which were categorized as high-quality studies.The principal features of each study were illustrated at Table 2 = 76.1%, P < 0.001), random-effects model was selected for measuring the association of miR-203 elevation and clinical outcome of CRC. The pooled analysis of the 11 studies revealed that elevated expression of miR-203 was demonstrated to be obviously correlated with shorter OS for CRC  , compared than Asian patients . The subgroup by specimens revealed the predictive role of miR-203 was especially more significant in tissue samples , but not in serum samples . Moreover, regarding the analysis of sample size subgroup, miR-203 exhibited a more prominent effect on predicting poor OS of CRC in large sample size  compared with small sample size .For explaining the heterogeneity in OS, subgroup analysis was further employed according to three subcategories containing sample source, sample size and the patient origin Table . When stP = 0.080), participant numbers (P = 0.078), and specimen sources (P = 0.993).Furthermore, a meta-regression was also performed to seek the possible sources that may be responsible for heterogeneity. The results revealed that the following factors may potentially cause the heterogeneity but did not reach statistical significance: patient origins , the module consisted of 40 nodes and 383 edges were identified to be the most significant. The nodes involved in the screened module were conducted function and pathway enrichment analysis, which were significantly involved in the pathways in cancer, proteoglycans in cancer, adherens junction, TNF, toll-like receptor, FoxO and colorectal cancer signaling as well as some other cancers including bladder cancer and pancreatic cancer  with a The above clinical data synthesis part has suggested the usefulness of miR-203 expression as a promising biomarker in predicting the survival outcomes for CRC. However, the mechanisms how miR-203 affected the initiation and progression of CRC was not fully understood. The molecular pathogenesis of CRC mainly comes from multistep, multifactor, and polygenic effects and involves changes in various oncogenes and tumor associated microRNAs. Thus we applied several bioinformatics methods for exploring the biological function of miR-203 at the functional level. Since microRNAs play the biological role by regulating downstream genes, we first conducted a functional enrichment analysis including GO and KEGG analysis with the regulated genes of miR-203. The GO analysis demonstrated that the genes regulated by miR-203 were primarily involved in regulation process for the biological process level, mainly enriched in basic cellular component for the cellular component level and significantly clustered in binding function and transcriptional activator activity for the molecular function level. KEGG pathway analysis indicated that some of the signaling pathways regulated by the target genes of miR-203 were involved in some important pathways. The majority of these signaling pathways have been demonstrated significantly related to the occurrence and development for CRC. For example, pathways in cancer may be the most crucial pathway due to the coverage of a series of pivotal signaling, for example the p53, TGF-β, Wnt and MAPK signal pathway, which have dominant impact on almost all the aspects of the tumorigenesis and progression of cancers including CRC. Accumulating evidence suggests that Wnt pathway is important during the pathogenesis of various types of cancers including CRC through influencing cellular proliferation, differentiation, and motility . The abnAs microRNA regulates multiple target genes, and each target gene has close connections with others. So we carried out a PPI analysis of genes regulated by miR-203 to explore more valuable information about miR-203. Next, based on the PPI network construction and analysis, top ten crucial genes were obtained. The crucial genes were demonstrated closely associated with some important pathways, which had pivotal impact on the occurrence and progression according to literature exploration. The most important module of the network map was further identified. The KEGG pathway analysis revealed that proteins of the prominent module were significantly enriched into a series of vital pathways. Most of the pathways have been confirmed by previous literatures. Here, we have to mention some more important pathways. It is worth noting that TNF, as an extraordinarily pleiotropic cytokine, is highly associated with maintenance and homeostasis of the immune system, inflammation as well as host defense . AdherenRecently, some studies on the tumor suppressive functions of miR-203 have been investigated. For example, recently, You et al.  reportedTo the most of our knowledge, our study may be the most comprehensive study about miR-203 as we not merely quantifies the biomarker performance of miR-203 for predicting the prognosis of CRC by meta-analysis, but also qualitatively explores the potential function and mechanism of miR-203 through integrative bioinformatics analysis. On the whole, our quantitative results strongly agreed with the present mainstream viewpoint that an undesirable impact of miR-203 high expression was related to worse clinical outcome. Moreover, there are still numerous valuable implications from our interesting results for future clinical and scientific research. In addition, although we have not conducted experiment, most of the enriched modules, pathways or networks were successfully confirmed by recent experimental literatures.However, several limitations exist in the present study. Firstly, the number of studies was relatively small. More clinical studies should be conducted to extend and validate the conclusions in the future. Secondly, we collected and pooled data from public studies rather than individual case data, so subgroup and regression analyses focusing on some important variables containing age, sex and stage of cancer were restricted due to the insufficient data. Thirdly, we only enrolled patients from Asians and Caucasians; so further studies assessed the biomarker roles of miR-203 about more ethnicities are required as the study population are not comprehensive. Besides, language bias may exist as only English studies were enrolled for evidence synthesis which may published more positive results. Due to the fact that all studies that are used for the data pooling have patients of various TNM stage (from stage I to stage IV), there is considerable heterogeneity in survival. Without patient-level data, it would be very difficult to tell whether miR-203 is actually useful as a prognostic factor. Finally, though we have validated the relevance of miR-203 status with the prognosis of CRC and proved the accuracy of our predictive results through an integrated bioinformatics analysis, well-designed biological experiments are still required for future confirmation as the initiation and development of CRC is complex, with multiple cumulative genetic changes.It was found that up-regulated miR-203 level may be a risk factor for CRC prognosis revealing a poor OS. Some vital genes, pathways and modules were identified through an integrated bioinformatics analysis, indicating the potential mechanisms of miR-203 during the initiation and progression of CRC. More large-scale and multicenter prospective clinical studies and biological experiments should be performed before the practical implementation for miR-203 regarding the survival prediction of CRC patients.


Acute kidney injury (AKI) is a common finding in patients with coronavirus disease 2019 (COVID-19) and has been associated with higher rates of death when compared to COVID-19 patients without kidney injury. Whereas the definitive pathogenesis of COVID-19-related AKI (CoV-AKI) is not clear, histopathologic evidence seems to point at multiple etiologies for the disease, including indirect and direct viral kidney injury. The high incidence of CoV-AKI, along with the aggressive clinical presentation of this entity, have increased the demands for kidney replacement therapies, rapidly overwhelming the supplies of healthcare systems even in major tertiary care centers. As a result, nephrologists have come up with alternatives to maximize the efficiency of treatments and have developed non-conventional therapeutic alternatives such as the implementation of acute peritoneal dialysis for critically ill patients. The long-term implications of CoV-AKI are yet unknown, though early studies suggest that around one third of the patients who survive will remain dependent on kidney replacement therapy. Nephrologists and healthcare workers need to be familiar with the clinical presentation and therapeutic challenges of CoV-AKI in order to develop strategies to mitigate the burden of the disease for patients, and for services providing kidney replacement therapies. It is estimated that the number of fatal cases of coronavirus disease 2019 (COVID-19) will near 500,000 in the United States (US) by the time this review is published . Acu. Acu8]. In general, the pathogenesis of CoV-AKI includes two major causes: (1) indirect injury resulting from the inflammatory and hemodynamic changes during COVID-19 and (2) direct injury induced to the organ by SARS-CoV-2.An uncontrolled inflammatory state induced by SARS-COV-2 infection and the resultant hemodynamic instability are likely to account for the majority of the cases of CoV-AKI. It is not coincidental then that most of the kidney biopsy series available for COVID-19 patients report variable degrees of classic ATI. AKI in COVID-19 may then follow similar pathogenic models to sepsis-induced AKI.A histopathological analysis on 26 autopsies in patients with COVID-19 described ATI in the proximal convoluted tubule (PCT) based on loss of brush border, vacuolar degeneration, and dilation of tubular lumen with cellular debris . A few oCytokine induced kidney injury is another proposed mechanism to the pathogenies of CoV-AKI. Several pro-inflammatory interleukins (IL), interferon (IFN), colony stimulating factors, tumoral necrosis factors (TNF), and chemokines are all potential mediators of AKI. The term cytokine storm was originally used to describe a syndrome of immune dysregulation that developed after chimeric antigen receptor (CAR) T-cell therapy . CytokinOrgan crosstalk may also play a role on the development of kidney injury in patients with COVID-19 . SeveralSARS-CoV-2 uses the angiotensin converting enzyme 2 (ACE2) receptor to enter cells . The virIn vivo and ex vivo evidence of kidney tropism from SARS-CoV-2 is conflicting. Early evidence suggesting presence of SARS-CoV-2 in the kidneys came from Farkash et al. after adverting in the tubular epithelium particles which were morphologically identical to SARS-CoV-2 in the autopsy of a patient who died from COVID-19 . However2 except for one patient. The investigators also found viral RNA and protein by using in situ hybridization (ISH) and indirect immunofluorescence (IF) in glomeruli and tubules. Kidney tropism was also corroborated by Braun et al. in a cohort of 63 patients [p = 0.003) [In a series of autopsies from COVID-positive patients, Puelles and collaborators detected viral load in kidney tissue using quantitative polymerase chain reaction (PCR) specific for SARS-CoV-2 in 13 out of 21 cases . The samOn the other hand, a study performed by Kudose et al. utilized immunostains for viral spike and nucleocapsid phosphoproteins, ISH for viral RNA, and ultrastructure examination with electron microscopy to test for kidney tropism from SARS-CoV-2 in a series of 17 patients. Their techniques did not detect virions or viral RNA in the kidney . One of It is still unclear if the nephron is a direct target of SARS-CoV-2, and though several models may potentially be used to explain cellular damage to the kidney parenchyma, these are all speculative. If confirmed, the presence of the virus in kidney cells may induce a local immune response and complement activation, ultimately resulting in inflammation, cell lysis, and endothelial damage with thrombi formation . Entry tA consequence of the variability in the pathogenesis of CoV-AKI is expecting similar variability in the clinical phenotype of this entity. Volume-responsive prerenal AKI may account for around 9% of the cases based on the report by Mohamed et al . The samUrinary abnormalities such as proteinuria and hematuria are very common in patients with SARS-CoV-AKI. Among 161 cases of CoV-AKI in New Orleans, de novo sub-nephrotic range proteinuria occurred among 39% of patients, and de novo nephrotic range proteinuria among 4% . ProteinIn the abovementioned series, there are conflicting data in regard to urinary sodium. While Mohamed et al. describe low urinary sodium in only 38% of the cases, Hirsch reports low urine sodium in 65.6%. It is worth mentioning that Hirsch and collaborators used a higher cutoff for low urinary sodium (less than 35 meq/L), whereas Muner used 20 meq/L. This may potentially help to explain why there is a large discrepancy among the two studies.Given the heterogeneity in the presentation of CoV-AKI, it is important to identify urine abnormalities in order to discern which patients may have AKI mediated by hemodynamic changes, and which patients may be presenting with glomerular injury and may benefit from a kidney biopsy. Limitations for the interpretation of existing data from urine studies in patients with COVID-19 AKI include the absence of serial urine measurements, especially in those patients with low urinary sodium, and the absence of separation of samples collected after indwelling catheter, which may have cause hematuria secondary to trauma.Susceptible individuals may be prone to the development of glomerular diseases during COVID-19. Collapsing glomerulopathy is the most common histopathologic finding in terms of glomerular diseases in patients with COVID-19. Patients may have different presentations ranging from isolated proteinuria in nephrotic ranges to AKI. While the pathogenesis is unclear, this feature is associated with other viral infections such as human immunodeficiency virus (HIV), Epstein–Barr, and parvovirus B-19 . The proThe immune response elicited by the viral infection has been associated with a hypercatabolic state similar to that seen in patients with rhabdomyolysis or tumor-lysis syndrome. Skeletal muscles loss has been linked to pro-inflammatory cytokines such as IL-6 and TNF-α ,76, whicPatients with CKD, especially those with end-stage kidney disease receiving periodic KRT, are a vulnerable population for adverse outcomes from COVID-19 in light not only of their kidney condition, but also of comorbid conditions. Several studies in the US reported mortality up to 30% for this patient population .To date, the largest study looking at the outcome of individuals with pre-existing kidney disease and COVID-19 comes from Flythe et al. on behalf of the STOP-COVID investigators . This grPatients receiving chronic in-center hemodialysis are at particularly high risk for SARS-CoV-2 exposure and infection given the inability to fully isolate . A largeOn the other hand, patients receiving chronic PD have a much lower risk of SARS-CoV-2 infection given the limited need for visits, now more than ever restricted thanks to the implementation of new technologies which allow for telehealth to be a feasible option. Among 818 patients on chronic PD who were followed from January to April in Wuhan, China, only 8 were diagnosed with COVID-19 .Now more than ever, it is imperative that nephrologists advocate for home-based dialysis in order to reduce SARS-CoV-2 exposure. Knowing that there are restrictions to candidature for home dialysis modalities, it is mandated for dialysis facilities to promote the safety of those who choose or already receive in-center hemodialysis. The effects SARS-CoV-2 infection has on individuals with immune-mediated glomerulonephritis (GN) were studied by Waldman et al. in the IRoc-GN international registry . In thisRecent guidelines were published by the Immunology Working Group of the European Renal Association—European Dialysis and Transplant Association (ERA-EDTA) for the management of patients with immune-mediated kidney disease during the SARS-CoV-2 pandemic. Among other recommendations, the group suggests to consider stopping or at least reducing the dose of antimetabolites, and postponing infusion of biological drugs in patients with COVID-19. Steroids should not be discontinued abruptly without hydrocortisone replacement . Kidney transplant recipients may be at particular risk for adverse outcomes from COVID-19 given the use of chronic immunosuppression along with the comorbidities which may be present in this population. The Montefiore Medical Center in New York City was the first to share their experience on 36 kidney transplant recipients diagnosed with COVID-19 . The autIn addition to augmented case fatality rates, several reports have also noted the overall increase in major adverse outcomes such as admission to the ICU and need for mechanical ventilation. A systematic review on 12 studies and 204 kidney transplant recipients diagnosed with COVID-19 reported that 34% of the patients in the review were admitted to the ICU, while 19.7% required mechanical ventilation . The largest dataset looking at the outcomes of kidney transplant recipients with COVID-19 comes from the European Renal Association COVID-19 Database (ERACODA). Their study enrolled 305 kidney transplant patients with COVID-19, 89% of whom required hospital admission. The 28-day probability of death within this group was 21.3%. In this study, the ICU admission rate was 21%, and 18% of the total patients required ventilatory support . The high incidence of CoV-AKI in the US imposed a significant burden on nephrology departments and ancillary staff. The challenge of providing adequate KRT to a large number of patients in the absence of adequate personal protective equipment (PPE), trained personnel, and dialysis equipment  prompted some institutions to make changes to their protocols and to adapt to this challenging scenario.Given the lack of strong data on how to treat CoV-AKI, management strategies have been extrapolated from non-COVID AKI studies. Clinical trials such as the IDEAL-ICU , and STAThe same uncertainty applies to the selection of the preferred modality of kidney replacement therapy. In critically ill patients, there is no significant difference in patients’ outcomes between continuous kidney replacement therapies versus intermittent kidney replacement therapy. Therefore, different modalities are chosen based on institution availability and patient tolerability. Equally to non-CoV-AKI cases, continuous kidney replacement therapies are the modalities of choice in patients with hemodynamic instability who are not able to tolerate intermittent hemodialysis (iHD). No head-to-head trial has been made comparing the different modalities of continuous kidney replacement therapy (CKRT) in CoV-AKI.There are no solid data to suggest that removal of pro-inflammatory cytokines through convection will improve treatment outcomes or increase survival. There are only small case reports and case series to support the use of extracorporeal therapies or highly adsorptive filters to remove pro-inflammatory molecules. The rationale of removing pro-inflammatory molecules by ultrafiltration seems reasonable, but further studies will be needed in order to assess the potential benefits of these therapies.Acute PD is a reasonable option to offer continuous or intermittent kidney replacement therapy as an alternative to hemodialysis or hemofiltration. Peritoneal dialysis is rarely used as an option for hospitalized patients with AKI . HoweverIn the face of shortage of KRT machines and human personnel capable of providing therapy, institutions had to make adjustments to standard treatment practice. Higher dialysate flow rates were used in continuous kidney replacement therapy modalities when treatment time was decreased, and additional extension tubing was used in order to place dialysis and CKRT machines outside patient rooms in order to minimize exposure to COVID-19 for doctors and nurses. Isolated slow continuous ultrafiltration sessions were implemented in between treatments for volume management if diuretics were not effective ,100. Simp = 0.001). Patients receiving SLEDD seemed to benefit from low-molecular-weight heparin (LMWH) over unfractionated heparin or the direct thrombin inhibitor Argatroban .A particular complication noted in critically ill patients with COVID-19 ARDS is frequent vascular access and dialysis filter clotting. An early retrospective study from Wuhan alluded to the high mortality rate in COVID-19 patients who had abnormal coagulation parameters, compared to those with normal parameters . In a coAnother retrospective study conducted in New York City took a different approach by looking at the life of 502 CKRT circuits on different anticoagulation regimens . ClottinFurther studies need to be conducted in order to assess for optimal strategies in anticoagulation for patients with CoV-AKI receiving KRT. Despite disparities in outcomes among individual strategies, it is an undisputed fact that circuits without anticoagulation tend to perform poorly in patients with CoV-AKI when compared to anticoagulated systems. Anticoagulation options for patients on KRT are summarized in Kidney injury is a common extra pulmonary finding in patients with COVID-19. Though further research is still needed to understand the mechanisms of injury, it is likely that more than one mechanism is implicated. Despite the variability of reports on the incidence of CoV-AKI, we have to expect a high influx of patients and high demands that may overwhelm inpatient nephrology services. As such, it is imperative that centers prepare to match this demand by anticipating the need for human and non-human resources , and by developing strategies such as the creation of urgent peritoneal dialysis programs to assist hemodialysis services. Physicians also need to be aware of the special challenges in treating COVID-19-related AKI, especially in terms of KRT. There is hope that most of the survivors will have at least partial recovery of kidney function, though we should be preparing for a raise in the incidence of CKD and ESKD.


A total of 80 participants were recruited from three private middle SES schools of Rahim Yar Khan, Pakistan to explore the narrative skills development in Urdu-speaking preschoolers aged between 4-and-5 years. Data collection was completed using a two-stage sampling technique (convenience and purposive). After obtaining parental consent, the participants were screened for their intellectual functioning. Three personal narrative samples, using conversational maps, were collected from the participants who passed the screening. The narrative data were analysed on both macro and microstructural levels by using high point analysis and use of cohesions , mean length of utterances in words (MLUw), the total number of words (TNW), and the number of different words (NDW). The data presented here include age and gender of participants, their scores on the verbal IQ test, the scores obtained on macro-and microstructural levels. The results based on differences and relationships of the data obtained are published somewhere else  The data will help to form a baseline for future studies in the related domains.•All the researchers working in the field of language development not only from Pakistan where Urdu is the lingua franca but also from around the globe where the Urdu-speaking children reside as social ethnic minorities can benefit from this data.•The presented data can be further utilized to design studies based on the comparison of these 4-and-5-year olds with Urdu-speaking adults and children from different age groups. These data are based on typically developing (TD) children, henceforth, can also be used for comparing the macro-and microstructural skills of TD children with different developmental disorders, to find out the deviated patterns. Moreover, this data can also be used for cross-linguistic comparisons.1The attached excel file contains the demographic information of the participants including age and gender. In addition, it includes the verbal IQ scores, macrostructure scores analysed using High point analysis 22.1The between-group cross-sectional research design was employed to explore the patterns of personal narrative development in the Urdu-speaking preschoolers.2.2Eighty Urdu-speaking children were recruited by using the two-stage sampling technique from three middle socioeconomic-status (SES) schools of Rahim Yar Khan, Pakistan. The stage one employed the convenience sampling technique meaning that the children whose parents were available and were willing to provide the consent of participation were included. Then in stage two by applying the purposive sampling technique, the children were screened for their intellectual functioning, and those who passed the screening were included. Slosson Intelligence Test, Revised (SIT-R3) , and 5-year-olds . There were 38 girls and 42 boys in the final sample. Additional information based on the acquisition of language developmental milestones was obtained from the mothers within a demographic form (constructed by researchers) attached to the consent form.Based on the screening results, the final sample included two main groups with 40 children each, divided into groups based on their age, 4-year-olds 2.3The personal narratives were elicited using Peterson and McCabe's 2.4The narrative analyses were carried out both on macro and microstructural levels. For macrostructure, high point analysis by McCabe and Rollins 2.5All the statistical analyses were conducted using SPSS v21.The authors declare that they have no known competing financial interests or personal relationships which have, or could be perceived to have, influenced the work reported in this article.


ApoB48 is produced after post-transcriptional RNA modification by Apobec-1 editing enzyme, exclusively in the small intestine of humans and most other mammals. In contrast, in the liver where Apobec-1 editing enzyme is not expressed (except in rats and mice), the unedited transcript encodes a larger protein, ApoB100, which is used in the formation of very low-density lipoproteins (VLDL) to transport liver-synthesized fat to peripheral tissues. Apobec-1 knockout (KO) mice lack the ability to perform ApoB RNA editing, and thus, express ApoB100 in the intestine. These mice, maintained on either a chow diet or high fat diet, have body weight gain and food intake comparable to their wildtype (WT) counterparts on the respective diet; however, they secrete larger triglyceride (TG)-rich lipoprotein particles and at a slower rate than the WT mice. Using a lymph fistula model, we demonstrated that Apobec-1 KO mice also produced fewer CMs and exhibited reduced lymphatic transport of TG in response to duodenal infusion of TG at a moderate dose; in contrast, the Apobec-1 KO and WT mice had similar lymphatic transport of TG when they received a high dose of TG. Thus, the smaller, energy-saving ApoB48 appears to play a superior role in comparison with ApoB100 in the control of intestinal lipid transport in response to dietary lipid intake, at least at low to moderate lipid levels.Fatty meals induce intestinal secretion of chylomicrons (CMs) containing apolipoprotein (Apo) B48. These CMs travel  Obesity has become a global epidemic, affecting more than 39.8% of adults in the US and increasing risk of cardiovascular diseases (CVD) and stroke . Epidemi2 to yield lysophosphatidylcholine and FA in the intestinal lumen. After enterocyte uptake, most of the lysophosphatidylcholine is reconstituted to form phosphatidylcholine  and then combined with a B48-free lipid droplet to form a pre-CM . WhereasIntestinal TG-rich lipoproteins are essential for transporting dietary fats into the circulation . The secvia a nonproteasomal, post-ER, presecretory proteolysis (PERPP) pathway, likely involving autophagy, in conditions when TG availability and incorporation into VLDL are normal  pathway for destruction . When the normal .2 cells (a human colon carcinoma cell line), little or no ubiquitination and proteasomal degradation of ApoB48 synthesized in colon cell lines is observed  and CaCoobserved . In conterocytes . In vivoy lipids . WhetherGeneration of the Apobec-1 knockout (KO) mouse, which is incapable of ApoB48 synthesis, has offered a unique opportunity to study the importance of ApoB48 through comparison of CM formation and lipid transport in WT animals versus the Apobec-1 KO animals with exclusive ApoB100 production . The Apoin vivo within the small intestine, accounting for the decreased secretion of TG-rich CMs observed in our previous report  deg ApoB100 , but ther ApoB48 ; and (3)r ApoB48 . Althougr ApoB48 , its regIn an early study utilizing blockade of plasma lipoprotein degradation, it was shown that fat-fed Apobec-1 KO mice produced larger plasma lipoproteins with greater amounts of TG in CMs than WT mice . Since pDietary lipids are absorbed by intestinal enterocytes, and the transport of long-chain fatty acids from the small intestine to circulation is controlled primarily by a constant number of ApoB48-containing lipoprotein particles with alterable particle size . InsulinThe raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The animal study was reviewed and approved by the Institutional Animal Care and Use Committees at the University of Cincinnati. Written informed consent was obtained from the owners for the participation of their animals in this study.CL contributed to data collection and analysis and manuscript preparation. KC contributed to data analysis and manuscript preparation. 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.


O-sulfated galactose head group of sulfatides, an abundant glycosphingolipid class, poses the (sphinx-like) riddle on involvement of glycan bridging by tissue lectins (sugar code). First, synthesis of head group derivatives for functionalization of amphiphilic dendrimers is performed. Aggregation of resulting (biomimetic) vesicles, alone or in combination with lactose, demonstrates bridging by a tissue lectin . Physiologically, this can stabilize glycolipid-rich microdomains (rafts) and associate sulfatide-rich regions with specific glycoproteins. Further testing documents importance of heterobivalency and linker length. Structurally, sulfatide recognition by galectin-8 is shown to involve sphingosine's OH group as substitute for the 3′-hydroxyl of glucose of lactose. These discoveries underscore functionality of this small determinant on biomembranes intracellularly and on the cell surface. Moreover, they provide a role model to examine counterreceptor capacity of more complex glycans of glycosphingolipids and to start their bottom-up glycotope surface programming.The small 3-  •N)-glycan bridging by galectins-4 and -8Nanoparticle programming detects sulfatide-(•Protein design (linker/domain type) is a switch for aggregation activity•Sphingosine's OH group is involved in contact building with a galectin Supramolecular Chemistry; Biochemistry; Biophysics O-sulfated derivative in sulfatides has started efforts to explain the abundant presence of a lipid-linked monosaccharide and its site-specific sulfation. Of course, it is reasonable to assume that these simple compounds are more than inert constituents of the lipid bilayer . The occurrence of further sulfotransferases acting on sugars and their diversification to gain selectivity for diverse substrates are the pillars of the hypothesis for (patho)physiological relevance of glycan sulfation, which is assumed to convert rather small glycans (even a monosaccharide) into potent ligands  at branch ends of glycan chains after protein glycosylation is a substrate   present their own mysteries . Especiaglycans) . Howeverglycans) , as wellglycans) , 2016, tHere, we describe preparation of conjugatable headgroups and their alkyne-functionalized adapter for linking sulfatide derivatives to a lipid anchor to prepare custom-made amphiphilic Janus glycodendrimersomes (GDSs), which made galectin testing (incl. architecture variants obtained by protein engineering) possible: this strategic combination is applied as a step to solve pertinent mysteries both on (ga)lectin/glycosphingolipid presence and on structural diversity of galectins. The advantage of proof-of-principle work with amphiphilic Janus glycodendrimers, besides the perspective for chemical programming of the nanoparticle surface, is gaining access to diverse morphologies relevant for pathobiology such as cubosomes  or for gtrans-interactions by aggregation assays, we also probed into the structural basis of contact building on the atomic level. This process commonly requires extension of galactose to the disaccharide (lactose) for galectins so that the glucose moiety can contribute, as revealed for the galectin CRD by crystallography ). Its synthesis starting from galactopyranoside 1 . Their application in aggregation assays documented that both CRDs in the heterobivalent Gal-4 can associate to this type of ligand, hereby bridging the suLac (type I)-bearing GDSs  in such microdomains and glycoprotein cargo with LacNAc (or suLac) termini together physiologically.Interestingly, homodimeric (linker-free) Gal-1, -2 and -7 are active, pointing to a combination of nature of CRD and architecture of its presentation for extent of activity , right. 4 described in a patent by  or sulfatide-2   sulfatide presence in detergent-resistant membrane sections. Remarkably, these artificial nanoparticles established the assay platform to explore sulfatide activity as ligand in our study. Compared with suLac (type I), their ligand activity for Gal-4 was rather small but clearly detectable  will be a respective sensor. The inert sugar mannose (Man) serves as negative control  part of suLac(NAc) may play a significant role here, when noting the respective capacity of suLac as docking point. The two tandem-repeat-type galectins thus appear to use a compensatory contact for acquiring stability, and here sphingosine's hydroxyl group comes into play as a possibility. A means to test the validity of this hypothesis is crystallographic study, if a complex of a galectin CRD with synthetic sulfatide-1 could be obtained.Having revealed physical contacts in this heterotypic  system, the question can be addressed next as to whether Gal-4 is unique for this interplay. Respective testing disclosed aggregation also for Gal-8 . The los6Z6Y). Clear electron density for the head group structure was obtained, whereas the amide's acyl chain appeared rather mobile, so that a profiling of the head group's contacts became possible. As depicted in 3AP6), is illustrated in 5DUW, Systematic testing of conditions with the N-terminal CRDs of Gal-4 and -8 led to crystals for Gal-8N and thus data to decide the issue sphingolipids and glycerophosphatides typically bearing unsaturated acyl chains and addition of cholesterol are then helpful to constitute systems with a larger number of variables that can implement and regulate fluidity. Variation of acyl chain length, for example to the C24 form of sulfatide assumed to be the preferential binding partner , is likepaul.v.murphy@nuigalway.ie).Further information and requests for resources should be directed to and will be fulfilled by the Lead contact, Paul V. Murphy (This study did not generate new unique reagents.The published article includes all datasets/code generated or analyzed during this study.All methods can be found in the accompanying