The United States continues to be impacted by decades of an opioid misuse epidemic, worsened by the COVID-19 pandemic and by the growing prevalence of highly potent synthetic opioids (HPSO) such as fentanyl. In instances of a toxicity event, first-response administration of reversal medications such as naloxone can be insufficient to fully counteract the effects of HPSO, particularly when there is co-occurring substance use. In an effort to characterize and study this multi-faceted problem, the Camden Opioid Research Initiative (CORI) has been formed. The CORI study has collected and analyzed post-mortem toxicology data from 42 cases of decedents who expired from opioid-related toxicity in the South New Jersey region to characterize substance use profiles. Co-occurring substance use, whether by intent or through possible contamination of the illicit opioid supply, is pervasive among deaths due to opioid toxicity, and evidence of medication-assisted treatment is scarce. Nearly all (98%) of the toxicology cases show the presence of the HPSO, fentanyl, and very few (7%) results detected evidence of medication-assisted treatment for opioid use disorder, such as buprenorphine or methadone, at the time of death. The opioid toxicity reversal drug, naloxone, was detected in 19% of cases, but 100% of cases expressed one or more stimulants, and sedatives including xylazine were detected in 48% of cases. These results showing complex substance use profiles indicate that efforts at mitigating the opioid misuse epidemic must address the complications presented by co-occurring stimulant and other substance use, and reduce barriers to and stigmas of seeking effective medication-assisted treatments. Since the 1990s, the United States (US) has seen three waves of the opioid use epidemic, beginning with increases in toxicity cases involving prescription opioids, followed by heroin and, more recently, fentanyl and fentanyl analogs otherwise known as highly potent synthetic opioids (HPSO) . AlthougIn addition to the increase in the number of toxicity events involving HPSO, there has also been a nationwide increase in the number of toxicity deaths that involve both HPSO and stimulants , 13–15. Whereas an increasing prevalence of HPSO and a lack of toxicity preparedness are risk factors for opioid-related toxicity death, a more comprehensive understanding of drug toxicity risk is still needed. The Camden Opioid Research Initiative (CORI) is a multi-armed research study including a biobank resource of biospecimens collected from individuals who died from opioid-related drug toxicity (CORI Biobank) . As partSamples were collected from decedents in Camden or Gloucester Counties, New Jersey, US, between March 2019 and April 2021. Deaths evaluated and affirmed by the Gloucester County medical examiner’s office to have been due to opioid-related drug toxicity were considered for inclusion in the study. Inclusion criteria specify that the next of kin must be identifiable and able to be contacted, over age 18 years, not incarcerated, English language proficient, and agree in writing to the sample’s submission to the study while the decedent is still in the custody of the medical examiner; any sample not meeting these conditions was excluded.The study described in this protocol is not classified as Human Subjects Research because it involves the collection of blood samples after death; however, the CORI Biobank is overseen by the Coriell Institute of Medical Research Institutional Review Board (IRB) to ensure that the rights of the donor and their next of kin are appropriately respected. The study was conducted according to the guidelines of the Declaration of Helsinki, and the study protocol, #R163, was reviewed and approved by the Coriell IRB.TM Panel), to identify fentanyl, fentanyl derivatives, synthetic cannabinoids, synthetic benzodiazepines, and other synthetic opioids that may contribute to opioid-related toxicity deaths. The full list of 77 compounds detected in our cases by the toxicology panels, including their classification for purposes of analysis and reporting in this study, and data on detected sample concentrations is shown in After assessing the decedent’s death was due to opioid toxicity, the medical examiner collected a sample of femoral blood in two vials containing sodium fluoride (NaF). The vials were then shipped to NMS Labs  for expanded testing upon written consent from the decedent’s next of kin. Expanded toxicology testing at NMS involved two panels, 8092B (Forensic Postmortem Prescription Drugs Screen) and 8054B  and seven female decedents (17%). The median age of all decedents was 35.5 years with a range  of , 58 yearThe toxicology analysis revealed the presence of multiple compounds in postmortem blood samples from every decedent, with a median of nine compounds per sample (range 5–19). Eighteen of the compounds represent pairs of active drugs/analytes and respective metabolites, such as fentanyl-norfentanyl, where fentanyl is the un-metabolized HPSO, and norfentanyl is its metabolite. One compound, ethylecgonine, is formed when alcohol and cocaine are used simultaneously and was also reported in our screen . Table 1The compound class detected with the greatest frequency, in all 42 decedent samples, was stimulants, which includes commonly consumed substances of caffeine and nicotine, and though these are not substances typically considered subject to substance use disorder, they may contribute to an additive effect when combined with other simulants. The most common stimulant was caffeine, detected in 38 samples (90%), followed by nicotine and its metabolite, cotinine, detected in 34 samples (81%). Cocaine and/or its metabolites were detected in 13 (31%) of the samples.Next in frequency, the class of HPSO compounds including fentanyl and fentanyl metabolites were found in 41 (98%) of 42 postmortem blood samples. Median concentration of un-metabolized fentanyl was 16 ng/ml (detected range 1–73 ng/ml) and median concentration of norfentanyl was 2 ng/ml (detected range < 1–20 ng/ml). Other non-fentanyl HPSO were detected in seven (17%) of the samples, and free morphine was detected in 14 (33%) of the samples; of the 14 samples that tested positive for free morphine, nine registered sub-median fentanyl concentrations. The one case with no detectable fentanyl or fentanyl metabolites had a particularly high concentration of methadone. Opioids not classified as HPSO, which include codeine, morphine, and oxycodone, were detected in 16 (38%) of the samples as the fourth most frequent class of compounds, followed by non-opioid analgesics such as ibuprofen and acetaminophen in 15 (36%) of the samples. Antidepressant and antianxiety compounds were detected in 13 (31%) of the samples.Medications to counteract opioid toxicity or to treat opioid addiction were among the least frequently detected compound classes. The opioid antagonist, naloxone, used to reverse the effects of opioid toxicity, was detected in eight (19%) of the samples. Available data on the decedents does not include the time at which naloxone had been administered relative to intake of the fatal dose of opioids or to the time of death. Most strikingly, just three (7%) of the samples showed evidence of medication used in the treatment of OUD (MOUD), which include methadone and buprenorphine. The MOUD, buprenorphine, was detected in one of these three samples, and methadone in the other two samples. One sample testing positive for methadone, also the lone sample without evidence of fentanyl, showed a high methadone concentration of 1400 ng/ml. Death due to methadone toxicity has been reported to occur at a concentration of 957 ng/ml, and the high concentration of methadone may have contributed to the fatality in this case, given the absence of HPSO .Highlighted by the ubiquitous presence of stimulants in postmortem blood among the donors to the CORI biobank, the toxicology results document a high prevalence of co-occurring substance use profiles that accompany opioid use. Specifically, among commonly-recognized substances subject to abuse, 13 (31%) of the samples tested positive for cocaine, any sedative including benzodiazepine anxiolytics were found in 13 (31%), other benzodiazepines in seven (17%), amphetamine and/or methamphetamine in six (14%), alcohol or alcohol metabolites in eight (19%), and cannabinoid compounds in seven (17%) of the samples. Co-occurring substance use was prevalent in both sexes. Stratification of the toxicology data by sex or age did not reveal statistically significant differences between male and female or young and old decedents . WithinConsidering only stimulants, sedatives, dissociatives, and psychoactive bath salts as those compound classes associated with SUD, while excluding fentanyl and HPSO, non-HPSO opioids, as well as commonly-consumed caffeine, nicotine, and its metabolite, cotinine, from the stimulants, a large majority of 36 (86%) of the samples showed a profile of co-occurring substance use, while just three (7%) of the samples showed evidence of MOUD in a possible effort to treat opioid use disorder. The prevalence of co-occurring substance use and the low rate of MOUD observed in our cases are consistent with literature reports on individuals with OUD . For exaAmong the stimulants, cocaine and its analytes and metabolites were detected in 13 31%) of the cases. Cocaine can lead to relapse and attrition from treatment programs for OUD . Account% of the The high incidence of caffeine, a stimulant also metabolized by cytochrome P450 enzymes, detected in 37 (88%) of the samples, may be due to commonly consumed caffeinated beverages, though there is literature to support that caffeine is introduced into the illicit opioid supply in an effort to make the drug less lethal in addition to simply adding bulk for increased profit –37. AccoFentanyl was the dominant HPSO among this class of compounds in the CORI Biobank study. In the past decade, deaths from opioid-related toxicity have been driven by an increase in HPSO use , 39, a tPresuming that fentanyl is a likely causative factor in most of these drug toxicity deaths, the lethal dosage is difficult to estimate given the uncertain timing of fentanyl exposure relative to the time of toxicity reaction and to the pharmacokinetics for chronic use or OUD. Short-acting opioids are cleared from the bloodstream in 2–4 days; however, the metabolism and clearance of fentanyl, particularly in individuals exposed to opioids chronically, require a much longer time . For exaAmong sedatives, the third most common class of compounds detected after stimulants and HPSO in the CORI Biobank, benzodiazepines were detected in 17% of cases, and though there are reports of this class of drugs appearing in the illicit opioid supply and in postmortem samples from drug-related deaths, it is unclear whether the benzodiazepine was combined with an opioid or taken separately , 50. XylMedications commonly used to treat depression and psychiatric conditions were detected in 12 (29%) of the cases, though it is unclear if the antidepressants were obtained from a physician as a measure of clinical care, as medical records were not available to the researchers of this study. Although antidepressants are subject to misuse by those with OUD, they also associate with increased retention to buprenorphine treatment for OUD , 56. HowTo test the hypothesis that individuals with OUD who are in clinical care may present with a lesser degree of co-occurring substance use than those who are not in clinical care, we compared the average number of putative illicit substances besides opioids  between decedents with and without evidence of antidepressant/antianxiety medications in their toxicology results. However, there was no significant difference in the average number of non-opioid illicit substances detected between the two groups .Our results show some evidence that opioid-toxicity reversal medication was administered to decedents, although the timing of the medication dose with respect to the toxicity reaction is unclear. Naloxone, often used in attempt to reverse an opioid toxicity reaction at the time of first response, was detected in eight (19%) of the cases. The overall efficacy of naloxone against fentanyl and fentanyl analogs is questionable given the rapid onset time of toxicity for HPSO and the very brief time window for reversal of toxicity events . AlthougBuprenorphine and methadone, currently approved MOUD treatment drugs, were detected in only three of the decedent toxicology samples. The small number of cases with evidence of MOUD (7%) lends support to concerns about limited access to medication treatment and clinical care for those living with OUD, and about underuse and low retention rates of MOUD particularly among those who engage in co-occurring substance use , 65, 66.Potential explanations for the low incidence of MOUD in this study are limited access to MOUD, low adherence rates for MOUD, and the stigma of seeking treatment –77. WhilThe low incidence of detected MOUD, buprenorphine or methadone, in the current study may be explained because these drugs protect against death from opioid toxicity when taken at the correct dosage. Indeed, prior studies show that MOUD significantly decrease the occurrence of toxicity events among persons with OUD, with additional reductions in the likelihood of toxicity events associated with longer durations of medication treatment for OUD up to 12 months . BuprenoExpanded toxicology reports of 42 decedents who expired from opioid-related toxicity in the South New Jersey region from March 2019 through April 2021 were analyzed as part of the Camden Opioid Research Initiative. The HPSO, fentanyl, was found in 41 (98%) of the toxicology cases, and either buprenorphine or methadone was found in only three (7%) of the cases, affirming a high incidence rate of opioid-toxicity events involving HPSO and a low incidence rate of MOUD among individuals who die from suspected HPSO toxicity. Naloxone, a toxicity reversal medication to be administered at the time of first response, was detected in eight (19%) of the cases. The outcome suggests that the naloxone dose may have been insufficient to counteract the HPSO or the HPSO when combined with an array of other detected compounds, as evidence of co-occurring stimulant use was detected in all decedent samples, and sedative use in nearly half of decedent samples, often coinciding with non-HPSO opioids and non-opioid analgesics. Alternatively, failure to detect naloxone in some samples may be due to its short half-life.To confront the increased risk of opioid toxicity death presented by HPSO and co-occurring substance use, a multi-faceted strategy is needed. Guided by the data observed in this study, an effective response to reduce the likelihood of a fatal toxicity event would include special consideration for co-occurring substance use in both treatment and emergency response with respect to naloxone dosing, increased and immediate availability of naloxone to the general population, elimination of MOUD stigma, and increased access to MOUD and clinical care.S1 FigFrequency of toxicology compounds by class, in descending order, detected per case in 42 postmortem samples of femoral blood collected from opioid-related toxicity cases submitted to the CORI Biobank. Each bar on the x-axis is a detected compound class, the y-axis is the case incidence count, and the shading indicates the number of compounds detected for a given class per case.(TIF)Click here for additional data file.S1 TableCompounds (N = 77) detected in expanded toxicology panels among postmortem blood samples collected from 42 opioid-related drug toxicity death cases as part of the Camden Opioid Research Initiative. Detected concentrations are shown numerically where available, else, reported as Positive.(DOCX)Click here for additional data file.S1 FileA full listing of the more than 360 analytes tested by the two panels, 8054B and 8092B, at NMS Labs.(XLSX)Click here for additional data file.S2 FileDouble-blinded raw data of toxicology testing results used for analysis in this study.(XLSX)Click here for additional data file.


This study aims to examine the time-varying efficiency of the Turkish stock market’s major stock index and eight sectoral indices, including the industrial, financial, service, information technology, basic metals, tourism, real estate investment, and chemical petrol plastic, during the COVID-19 outbreak and the global financial crisis (GFC) within the framework of the adaptive market hypothesis. This study employs multifractal detrended fluctuation analysis to illustrate these sectors’ multifractality and short- and long-term dependence. The results show that all sectoral returns have greater persistence during the COVID-19 outbreak than during the GFC. Second, the real estate and information technology industries had the lowest levels of efficiency during the GFC and the COVID-19 outbreak. Lastly, the fat-tailed distribution has a greater effect on multifractality in these industries. Our results validate the conclusions of the adaptive market hypothesis, according to which arbitrage opportunities vary over time, and contribute to policy formulation for future outbreak-induced economic crises. Financial crises are one of the primary causes of financial asset depreciation. For example, the global financial crisis (GFC), which began in the United States’ housing market, first affected financial markets and then the real economy via financial channels  proposed by Fama  constituweak-form, semi-strong form, and strong form. The weak form of market efficiency indicates that investors cannot obtain abnormal returns from past asset price movements because market prices have already reflected all past and current information about price and volume. Meanwhile, the semi-strong form of market efficiency states that the market value of an asset adapts instantly to all new information in pursuit of a random walk process from past prices. Lastly, the strong-form market efficiency hypothesis holds that a single investor cannot make abnormal profits even with insider information  model to assess and compare aggregate and sectoral market efficiency during times of crisis. Kantelhardt et al.  proposedThere are numerous studies in the literature that investigate the market efficiency of Borsa Istanbul. Some studies  behavior of the emerging (developed) Islamic stock market is inconsistent, whereas the short-run (long-run) behavior of the developed Islamic stock market is consistent. Han et al.  utilizedThe disagreement over whether the stock market is fully or adaptively efficient has increased academic interest in the AMH. Several studies have been conducted to investigate the effects of the AMH on stock markets. Ghazani and Araghi  estimateMandelbrot  and PeteSome studies have investigated market efficiency on a sectoral level. For instance, Cheong  examinedWe consider daily data, including the Borsa Istanbul (XU) 100 (BIST 100) index and the eight sector indices, which are industrials, financials, services, information technology, basic metals, tourism, real estate investment, and chemical, petroleum, and plastic. Stock return multifractality is frequently used as a measure of market inefficiency  properties in non-stationary financial time series and to determine the financial market efficiency  as follows (Wang et al. If the financial market is efficient, all small and large fluctuations follow a random walk process. In an efficient market, the MDM values will be equal to zero. In an inefficient market, however, the so-called value is higher. Tables The primary motivation for this study is to show which sectors have higher efficiency during various crisis periods. Market efficiency is an important indicator of whether or not an investor can profit from a return. In fact, some investments remained significant during the crisis periods. Depending on the motivation, we explore the time-varying market efficiency for sectoral markets using the MF-DFA to understand the features of the market periods, such as structural revolutions throughout the GFC and COVID-19 outbreak. Therefore, this study aims to contribute to the literature on AMH by performing the MF-DFA on aggregate and sectoral markets in Turkey during the GFC and COVID-19 outbreak.Our results have many intriguing implications for policymakers and investors. First, the findings show that all sectoral markets exhibit persistent behavior and multifractal characteristics, implying that arbitrage opportunities increased during the two crisis periods. The changing Hurst exponents at various time scales validate the AMH, in which arbitrage opportunities vary over time. The persistence levels of the sectoral markets differ between the two crisis periods, and speculative behaviors in all sectors are more prominent due to the features of each crisis, and follow a stronger behavior during the COVID-19 outbreak than during the GFC. This result supports the empirical findings of Choi  and MensSecond, the real estate and information technology sectors had the lowest efficiency during the periods of the GFC and the COVID-19 outbreak This finding is consistent with those of Choi  and CapoThird, due to the sovereignty of small fluctuations, all sectoral stock returns exhibit multifractality and long memory. During the GFC and COVID-19 outbreak periods, the real estate and industrial sectors have the strongest persistence, whereas the services and metals sectors have the weakest. It can be stated that previous period prices affect the formation of current prices in the real estate and industrial sectors due to their long memory features for the GFC period. In terms of policy, policymakers can enact regulations to increase market forecasting and attract various types of investors to these sectors. However, price changes in the services and metals sectors are not estimated by past price movements because they were the most efficient among the others during the COVID-19 outbreak.The findings reveal that convenient portfolio allocation, risk diversification, and hedging strategies may provide higher economic development. In other words, ensuring the sustainable development of the economy and sectors and the stock market revolution should be encouraged to maintain a dynamic and sustainable market. Moreover, market mechanism arrangements should be used rationally to decrease the effect of exogenous events on market efficiency. The investor structure in the stock market should be optimized, and the investment value of companies should be increased to provide relative benefits over financial intermediation. If the measures are implemented correctly, they will help boost market confidence, stabilize market sentiment, and encourage long-term improvement in the Turkish stock market.


Oncolytic viruses (OVs) for cancer treatment are in a rapid stage of development, and the direct tumor lysis and activation of a comprehensive host immune response are irreplaceable advantages of cancer immunotherapy. However, excessive antiviral immune responses also restrict the spread of OVs in vivo and the infection of tumor cells. Macrophages are functionally diverse innate immune cells that phagocytose tumor cells and present antigens to activate the immune response, while also limiting the delivery of OVs to tumors. Studies have shown that the functional propensity of macrophages between OVs and tumor cells affects the overall therapeutic effect of oncolytic virotherapy. How to effectively avoid the restrictive effect of macrophages on OVs and reshape the function of tumor-associated macrophages in oncolytic virotherapy is an important challenge we are now facing. Here, we review and summarize the complex dual role of macrophages in oncolytic virotherapy, highlighting how the functional characteristics of macrophage plasticity can be utilized to cooperate with OVs to enhance anti-tumor effects, as well as highlighting the importance of designing and optimizing delivery modalities for OVs in the future. Tumors often acquire immunosuppressive mechanisms through cancer immunoediting, which shapes the tumor microenvironment (TME) for tumor growth, effectively avoiding immune-mediated tumor clearance , 2. In fCurrently, tumor immunotherapy has received increasing attention with the focus on reducing tumor-associated immune evasion as the main therapeutic strategy for cancer treatment and depleting or “re-educating” cancer-promoting microenvironment cells into an immune-stimulating phenotype , 4, whicOncolytic virotherapy is an emerging immunotherapy, where oncolytic viruses (OVs) preferentially infect tumor cells to suppress tumor growth and are safe for other normal cells. Above this, OVs can activate the host immune system to generate an immune response to enhance anti-tumor efficacy , which cHowever, since OVs are essentially immunogenic infection factors, they stimulate the host to generate an immune response that can both produce antitumor effects and limit the activity of the virus itself . The delThe functional tendency of macrophages between OVs and tumor cells affects the overall therapeutic effect of oncolytic virotherapy, and how to eliminate the detrimental effect of macrophages on OVs and make them work in an anti-tumor direction is an important challenge nowadays. Therefore, in this review, we discuss the essential features and interactions between OVs and macrophages in antitumor immunity, and explore strategies by which to intervene with macrophages in oncolytic virotherapy to improve efficacy., including direct lysis of tumor cells, promotion of immunogenic cell death (ICD), induction of innate and adaptive immune responses, regulation of TME, and inhibition of tumor angiogenesis ) into liposomes coupled to chemokine CC motif ligand 2 (CCL2), which upon intravenous delivery binds to circulating monocytes expressing chemokine CC motif receptor 2 (CCR2), takes advantage of the aggregation of monocytes to hypoxic tumor vessels to deliver encapsulated OVs targeting tumor sites . This syTherefore, the use of carriers for adjuvant delivery of OVs is one of the promising strategies. This approach evades the capture of OVs by innate immune cells without affecting the body's immune function, while enhancing the targeting of tumors and reducing the viral delivery load.In conclusion, macrophages are an important factor affecting the therapeutic effect of OVs, and in the face of this dual effect, how to seek benefits and avoid harm is something we need to consider.In conclusion, the wide systemic distribution, plasticity and complex functions of macrophages make them largely influence the outcome and prognosis during oncolytic virotherapy. In the process of tumor development, M1 phenotype macrophages have anti-tumor effects and M2 phenotype macrophages have pro-tumor effects; in oncolytic virotherapy, macrophages can both enhance the effect of viral therapy and limit the spread of OVs to a certain extent. These different regulatory effects may be related to tumor heterogeneity, the type of OVs, and host responsiveness . The speFor the aspect of enhancing the beneficial effects of macrophages, gene-regulated OVs demonstrated a strong “re-education” capacity. Various cytokines, chemokines, and immunomodulatory antibodies remodel TAMs and enhance the anti-tumor effects of macrophages. This is one of the modalities with infinite exploits and possibilities that need to be constantly developed and added to.For the aspect of attenuating the harmful effects of macrophages, we need interventions to avoid unwanted immune responses; after all, intravenous delivery of OVs is a much more promising and ubiquitous mode of administration than local delivery. Direct strategies include the use of immunosuppressive or macrophage-depleting agents for short periods of time to inhibit macrophage capture of OVs. Indirect strategies include the use of genetic modification of viruses to avoid antibody-mediated macrophage clearance and the use of carrier cells or other nanomaterial encapsulation to overcome barriers to intravenous delivery.We also need to further examine the effect of antigen expression of various viruses on the activation of host immunity, and modification of OVs with extremely low antigen expression without loss of oncolytic biological activity is a major challenge for intravenous delivery. Therefore, we had to elucidate more deeply the interactions among OVs, macrophages, and tumor cells and their mechanisms of action, and utilize the powerful regulatory function of macrophages to synergistically enhance the application of OVs. In future clinical trials of oncolytic virotherapy, it is particularly important to optimize intravenous delivery, avoid unnecessary immune effects, reduce the delivery load of viruses, improve the oncolytic effect of OVs, and enhance the combined therapeutic effect to ultimately achieve a durable and effective treatment outcome.


Mitochondria are central hubs for energy production, metabolism and cellular signal transduction in eukaryotic cells. Maintenance of mitochondrial homeostasis is important for cellular function and survival. In particular, cellular metabolic state is in constant communication with mitochondrial homeostasis. One of the most important metabolic processes that provide energy in the cell is amino acid metabolism. Almost all of the 20 amino acids that serve as the building blocks of proteins are produced or degraded in the mitochondria. The synthesis of the amino acids aspartate and arginine depends on the activity of the respiratory chain, which is essential for cell proliferation. The degradation of branched-chain amino acids mainly occurs in the mitochondrial matrix, contributing to energy metabolism, mitochondrial biogenesis, as well as protein quality control in both mitochondria and cytosol. Dietary supplementation or restriction of amino acids in worms, flies and mice modulates lifespan and health, which has been associated with changes in mitochondrial biogenesis, antioxidant response, as well as the activity of tricarboxylic acid cycle and respiratory chain. Consequently, impaired amino acid metabolism has been associated with both primary mitochondrial diseases and diseases with mitochondrial dysfunction such as cancer. Here, we present recent observations on the crosstalk between amino acid metabolism and mitochondrial homeostasis, summarise the underlying molecular mechanisms to date, and discuss their role in cellular functions and organismal physiology. Mitochondria are the major sites for energy production. The tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) are coupled to metabolize energy sources for cellular function and survival. The close cooperation of these two processes ensures energy homeostasis under changing environmental and internal conditions. In addition to energy production, mitochondria are also hubs of cellular metabolism. A variety of metabolites from the TCA cycle are essential for cellular signaling and macromolecule synthesis . There iAmino acid metabolism is one of the most important sources for energy production in the cell. The metabolic pathways of amino acids are largely associated with mitochondria . In the etc.  in yeast and mammalian cells ; 2021. Mmembrane , for metmembrane . At the membrane   enriched in BCAAs show increased mTOR activity and mitochondrial biogenesis in skeletal muscle and hippocampus, as well as better physical and cognitive performance in aging compared to control-treated animals . Amino a of mTOR . In some of mTOR . Regulat of mTOR .The TCA cycle is a series of metabolic reactions that occurs in the mitochondrial matrix. The oxidation of fatty acids, glucose, and amino acids produces acetyl-CoA, and the TCA cycle uses acetyl-CoA to produce NADH and FADH2. NADH and FADH2 are then used for ATP production in the respiratory chain. The TCA cycle is not only involved in catabolism, but is also critical for the biosynthesis of a number of molecules including amino acids, lipids, and nucleotide bases . The comAlthough TCA cycle is at the nexus of both catabolic and anabolic metabolism involving different key fuel molecules and metabolites, it was recently reported that alteration of amino acid metabolism is the main consequence of TCA cycle disruption . Acute iMitochondrial OXPHOS is central to cellular energy production. The OXPHOS system includes five enzyme complexes and two mobile electron carriers (ubiquinone and cytochrome c) that drive the production of ATP . SimilarMitochondrial respiration in fission yeasts is also required for the provision of amino acids during fermentative propagation . InteresInhibition of the proteasome results in decreased levels of intracellular amino acids and causes mortality. Interestingly, the deleterious effect of proteasome inhibition can be reversed by amino acid supplementation . This fiSince mitochondria synthesize and degrade proteins independently, the question arises whether the availability of free amino acids affects protein turnover in mitochondria and how it influences the balance between mitochondrial and cytosolic proteins. Imbalance between mitochondrial and cytosolic proteins impairs mitochondrial function .via modulation of Opa1, an important molecule for inner membrane fusion, while starvation and OXPHOS activity regulate mitochondrial fission via modulation of Drp1, a GTPase recruited to OMM to mediate mitochondrial fission  are often associated with metabolic abnormalities. A detailed discussion of MDs has been reviewed recently . Here weSLC7A5), the major BCAA transporter, is highly expressed in many cancers, including lung cancer, prostate, and breast cancers (SLC1A4) is upregulated in both breast cancer and lung cancer (Glutamine metabolism plays an important role in both MDs and cancer due to its versatile role in cell metabolism . Cells w cancers . The serg cancer . In addig cancer . The rolg cancer . AlteratChanges in mitochondrial dynamics and functionality are often accompanied by metabolic adaptions, and cellular metabolic changes in turn lead to mitochondrial remodeling. Metabolism of most amino acids depends on the proper function of mitochondrial enzymes. Metabolic intermediates of amino acids are fed into the TCA cycle, and metabolites from the TCA cycle serve as precursors for amino acid synthesis. Dietary control or supplementation of amino acids has profound effects on the regulation of lifespan in various organisms, including worms, flies, and rodents. These age-related effects of amino acids occur directly or indirectly through the regulation of mitochondrial function. Certain amino acids such as alanine and BCAAs have been shown to be significantly elevated in primary MDs. Rewiring of the metabolic pathways of glutamine and other amino acids is thought to be an important bioenergetic adaptation in OXPHOS-defective cells or cancer cells. Recent studies have highlighted the role of the TCA cycle and respiratory chain activity in coordinating amino acid metabolism and mitochondrial homeostasis. Disruption of the TCA cycle or respiratory chain activity often has a reciprocal effect. Therefore, it is unclear whether many of the observed phenotypes are a direct consequence of inhibition of the TCA cycle or the respiratory chain. This should be considered in future studies. Dietary control or supplementation of amino acids regulates longevity, although the effects on longevity are contradictory in some cases. Dietary supplementation of most amino acids increases lifespan in worms, whereas restriction of essential amino acids extends lifespan in most studies in flies and mice. This may be due to the different preferred pathways of nutrient utilization in different organisms or to the dose and duration of amino acid supplementation. The sensitivity of different tissues to amino acid supplementation may also lead to different effects on lifespan. More detailed molecular mechanisms need to be characterized, particularly mitochondrial restructuring after amino acid treatment. In addition, the short- and long-term effects of amino acid supplementation on mitochondrial physiology, such as mitochondrial morphology and oxygen consumption, will be analysed. Furthermore, mitochondrial mass and metabolic activity vary in different tissues and cell types. Sensitivity to different mitochondrial gene mutations varies in different tissues, sometimes resulting in tissue-specific phenotypes and diseases . The int


In practice, the user creates a sample and uploads the Next-generation sequencing reads, then an analysis pipeline is launched automatically performing a series of typing and clustering operations fueling the information flow. Instances of IRIDA-ARIES host the Italian national surveillance system for infections by Listeria monocytogenes (Lm) and the surveillance system for infections by Shigatoxin-producing Escherichia coli (STEC). As of today, the platform does not provide tools to manage epidemiological investigations but serves as an instrument of aggregation for risk monitoring, capable of triggering alarms on possible critical situations that might go unnoticed otherwise.Pathogen genomics is transforming surveillance of infectious diseases, deepening our understanding of evolution and diffusion of etiological agents, host-pathogen interactions and antimicrobial resistance. This discipline is playing an important role in the development of One Health Surveillance with public health experts of various disciplines integrating methods applied to pathogen research, monitoring, management and prevention of outbreaks. Especially with the notion that foodborne diseases may not be transmitted by food only, the ARIES Genomics project aimed to deliver an Information System for the collection of genomic and epidemiological data to enable genomics-based surveillance of infectious epidemics, foodborne outbreaks and diseases at the animal-human interface. Keeping in mind that the users of the system comprised persons with expertise in a wide variety of domains, the system was expected to be used with a low learning curve directly by the persons target of the analyses' results, keeping the information exchange chains as short as possible. As a result, the IRIDA-ARIES platform ( The increasing application of Whole Genome Sequencing (WGS) in Public Health surveillance of infectious diseases, offers an excellent opportunity to employ the One Health approach  with theThe Italian National Institute of Health  deployed a genomic surveillance system for foodborne pathogens to shift from the existing typing system mainly based on the analysis of Pulsed Field Gel Electrophoresis (PFGE) profiles. This system is aimed at supporting the epidemiological surveillance of foodborne diseases in the population with specific short and medium/long term goals. The main short term goals were early detection of disperse outbreaks in the community, integration with genomic data from food/environment isolates to discriminate whether a certain food chain and vehicle is implicated or not in an outbreak. Likewise, integration of data and descriptive metadata from human and non-human isolates for source attribution and risk assessment studies were foreseen in the mid/long term to inform and evaluate the adoption of One Health control policies. This is particularly important for STEC control due to the large variety of hosts and sources that may play a role in the spread of infection to the most vulnerable population subgroups. For the purpose, the ARIES Genomics project planned to develop a platform as part of a One Health-Based Conceptual Framework , 3 startThe IRIDA-ARIES genomic surveillance information system is built engaging two open-source platforms: A Galaxy instance  implemenThe Galaxy Platform is a container for bioinformatic tools sharing a common workflow system, allowing each instance to focus on specific goals through the installation of appropriate tools. Each Galaxy instance is therefore different in function of the aims of the instances' managers. The code of the ARIES instance was installed as a role of the automation platform Ansible from the Galaxy Project  and was The Galaxy software is not suitable for the collection of samples with genomic and epidemiological data, nor is it possible to implement automation to the analyses. To this means, the open source IRIDA platform fitted the purpose, providing an intuitive web-based interface for the collection of genomics data, utilizing ARIES as a workflow engine for the bioinformatic analyses. In this scenario, IRIDA communicates with ARIES through the latter's unified Applications Programming Interface (API), hiding the ARIES platform from the user, who only interacts with the IRIDA user interface which was translated in Italian.The IRIDA software package being open source under the Apache License 2.0 was essential for the project because it allowed to fork (copy) the code and adapt the properties of the overall system. The system as a national surveillance platform had to be open to data obtained with various sequencing platforms, not only the mostly used Illumina paired-end reads but also Ion Torrent single reads. Development of bioinformatic tools in pathogen genomics is heavily biased vs. paired-end data. In the Galaxy platform it is not possible to create collections of single reads together with paired-end reads. The IRIDA software was therefore adapted to mask single reads as paired-end and the bioinformatic tools in ARIES were modified correspondently to intercept and elaborate them appropriately. Since this required a matching intervention in ARIES as well as IRIDA breaking functioning of the code, this change could not be opened as a pull request for IRIDA in order to synchronize this feature to the original upstream repository.Furthermore, since events of infectious diseases launched by European Union Member States on the European Centre for Disease Prevention and Control (ECDC) EpiPulse portal for the European surveillance of infectious diseases  frequentIn Italy, healthcare is delegated at the regional level comprising nineteen Regions and two Autonomous Provinces, where each has local health authorities and manages its proper surveillance systems in an independent way, hindering the acquisition of a nation-wide overview . To reflSeveral customizations have been introduced to automate as much as possible to lower the learning curve for unexperienced users while at the same time providing advanced tools for users with a genomics analysis background. The information flow is data-driven . To contAfter the automated workflow has concluded, an e-mail is sent to Project members containing concise information in function of the pathogen: the end-of-analysis message contains either core genome Multi Locus Sequence Typing (cgMLST) clustering results  or variant typing results. In case of a cluster involving more than one Region, the mail is also sent to any other Region involved, to the Ministry of Health and to the ISS to support coordination and outbreak management. A JSON file  containiAll analysis workflows have been designed specifically for the IRIDA-ARIES platform combining both existing as well as in-house developed tools. The workflows used for the automatic elaboration of samples are the most complex. The PHANtAsTiC  workflow [preprint ] has beeThe workflow is in its second version since the code has recently been adapted to match the cgMLST typing method performed at a European level by the European Food Safety Authority (EFSA) One Health WGS System . In factTo give users the possibility to further investigate selected samples within the system, several workflows have been added to the platform. These workflows comprise: cgMLST cluster analysis of the previously calculated allele profiles, reference-free Single-Nucleotide Polymorphism analysis using the PopPUNK software v1.1.2 , MinimumListeria monocytogenes based at the Abruzzo and Molise Veterinary Public Health Institute (IZSAM). Moreover, a tool is under development for the programmatic submission of cgMLST allele profile data to the EFSA One Health WGS system database. Locally, the associated analytical results of STEC data are visible in the web application of the NRL developed for the STEC collection.Sequences as well as metadata can be easily shared with other systems for further analyses. The IRIDA platform by default features a tool to assist in uploading sequence files to NCBI's Sequence Read Archive. A tool for the export of samples' metadata has been added to the platform. Currently, a collaboration agreement framework is in the process of being finalized, regulating the exchange of human and animal/food/feed Listeriosis data between the National Listeriosis Surveillance Working Group at ISS and the National Reference Laboratory (NRL) for The customizations to the platform have broken the encapsulation of the two underlying software packages. In fact, masking the heterogeneous data that is shared between them has limited the generality of both systems. Also, ARIES analysis workflows consume data directly from the IRIDA database.The IRIDA database is implemented on a single server but could be scaled up as a cluster. ARIES is relying on a SLURM  cluster The IRIDA platform is scaled up to four servers for high load deployment, dividing different tasks between them. With this configuration, batch uploads of several thousands of samples have been managed by the system. Currently, no further scaling of the system is possible.IRIDA-ARIES has to be considered as a component of the applications and protocols to be used in the ecosystem of surveillance, prevention and risk management. Its modular structure and the implemented APIs do allow for the flexible development of personalized interfaces vs. heterogeneous outputs.Although the platform is not designed to manage the whole process of surveillance and outbreak management, it comprises features for risk monitoring and is capable of automatically detecting clusters and triggering alarms on possible critical situations. Users are immediately aware of which Regions are involved in the warning and can readily establish connections while keeping information chains short. Regional data is shared to allow for a constantly updated national overview of pathogen diffusion. Feedback is returned to the regional users engaging them to participate actively with their data, creating a virtuous circle avoiding the danger of data silos at the regional level.Sharing of genomic data facilitates timely detection of clusters and, in general, situations of concern. Furthermore, the exchange with the veterinary public health Institutes  in a One Health view to receive human, animal, food and environmental samples, allows for direct comparison of genomic profiles in order to rapidly exclude possible contamination sources avoiding unnecessary high economic impact and to provide objective arguments to risk management for the timely activation of prevention measures. The exchange of sequence data without its metadata in case of suspect samples would avoid issues with data sharing. Should a situation of suspected outbreak occur, then an integrated data exchange protocol could be activated.Listeria monocytogenes and the local surveillance system for infections by Shiga toxin-producing Escherichia coli and counts 71 users, including personnel of the regional Public Health Services. For Listeriosis as of 14/12/2022, a total of 1,453 samples have been uploaded to the platform spanning the period 2002–2022, comprising 1,295 human samples, 61 animal/food/feed historical samples and 97 samples from outbreak events shared through European channels. The platform identified 108 clusters comprising 695 samples (73% of the clusters were composed of 5 or less samples). For STEC as of 14/12/2022, a total of 1,540 samples have been uploaded to the platform spanning the period 1989–2022, comprising 683 human samples, 798 animal/food/feed samples and 59 samples from outbreak events shared through European channels. In this case, 192 clusters have been identified by the platform including 664 samples (90% of the clusters consisted of 5 or less samples). Since PFGE typing was performed only in the presence of an epidemiologically identified suspect cluster and there was no collection of PFGE profiles from the territory, a comparison of cluster detection before and after the switchover is impossible.The IRIDA-ARIES platform is currently hosting the Italian national surveillance system for infections by The platform has been used to analyze the sequences of 42 STEC and 97 Lm isolates (accessed on 11/11/2022) appended to the information on the events of infectious disease, mainly outbreaks of infections, launched through the ECDC EpiPulse portal or to Urgent Inquiries launched on the former platform Epidemic Intelligence Information System for food- and waterborne diseases (ECDC-FWD-EPIS). The sequences were processed automatically by the platform upon upload and compared with the sequences of all the samples of the same species (for Lm) or serogroup (for STEC) isolated from human cases of disease in Italy already present in the database. This system was used to investigate 30 different events involving STEC strains and 71 involving Lm isolates, allowing to quickly reply on the ECDC FWD system about possible correlations among Italian isolates and those part of ongoing international events.Listeria monocytogenes and the Regions/Autonomous Provinces. The work of this Group supported the epidemiological investigation on the correlation between the clinical cases and the consumption of certain meat products. During the investigation, analysis of the cgMLST profiles allowed for the rapid identification of samples belonging or not to the specific cluster, narrowing the analytical process. The phylogenetic pipelines integrated into the platform have been used by the Working Group for the redaction of the periodic reports as well as autonomously by the regional users themselves. As stated by the Italian undersecretary of the ministry of Health in a parliamentary interrogation: “The current situation linked to Listeriosis has emerged thanks to the work of the Ministry of Health, through ordinary surveillance and through the IRIDA database of the Istituto Superiore di Sanità, which has made it possible to verify the increase in human cases throughout the national territory.”  and animal/food/feed (EFSA) concerning food- and waterborne diseases and zoonoses for a direct comparison with national samples, allows to integrate the Italian surveillance of foodborne diseases within an international One Health perspective. The objective is to align the typing workflows for each pathogen in collaboration with these European Agencies to obtain compatible results that can be readily exchanged.Listeria network has been foreseen, so the regional users get to know their counterparts from the other Regions, facilitating contacts in case of inter-regional clusters.In 2019, a face-to-face course was organized for the future regional users of the system. The feedback has been very positive, before the end of the course many participants had become confident with the system and acquired the ability to use most applications of the platform. Also, several requests from the participants could be readily implemented. A helpdesk has been set up to assist users running into problems. Now that the restrictions due to the COVID-19 pandemic have been largely lifted, an annual in-person meeting of the The system has been well-accepted by all different types of users because it has proven intuitive enough for those without specific computer skills, while yet powerful for the needs of the users with advanced bioinformatic experience. Although submission of data is on a voluntary basis in Italy, the system is now used by the majority of the Regions. Several clusters persistent in time and/or location have been highlighted by the system, indicating the platform as a powerful tool in support of future preparedness of early detection of food safety risks.Integration of human genomic data with samples originating from other One Health domains allows the platform to act as a key player in the surveillance of diseases caused by infectious agents in Italy. Not only in Italy though, the platform has been designed as multi-language and can readily be used in any (inter-)national context upon addition of a language dictionary. Issues with data sharing include data ownership, privacy regulations and legal considerations and have been tackled on several levels. The collection of the data has been approved by the Data Protection Officer of the ISS. The Regions remain owners of the genomic data they provide, their sequences cannot be accessed by others but only used in aggregated analyses.The platform has been used for STEC and Lm because the ISS already collected data for these pathogens and therefore the expertise for analyzing these genomes had been previously acquired. The surveillance of other pathogens could be implemented without much effort since the bioinformatic tools of the platform can be flexibly adapted. An IRIDA-ARIES instance named ICoGen , is actually in use at the ISS for the national surveillance of genomic variants of the SARS-CoV-2 virus.In the mid/long term, the IRIDA-ARIES infrastructure is meant to become the national platform for the genomic surveillance of infectious diseases. In this respect, the established networks providing data on Lm and STEC isolates from the different Italian regions, will be the starting point for expanding and consolidating the data providers' network for other foodborne infections. The analytical metadata of the sequenced strains will represent the central elements for the prompt identification of outbreak events as well as for source attribution and exposure risk assessment. Further development will focus on the integration of the platform as a component of an overall infrastructure for the surveillance and management of infectious diseases. The hope is that IRIDA-ARIES through the establishment of an inclusive cross-sector network will serve as a basis and stimulus for the creation of a national systemic approach enabling source attribution studies such as those carried out in the DiSCoVeR project  and posshttps://github.com/aknijn/irida PHANtAsTiC: https://github.com/aknijn/phantastic-galaxy.The datasets presented in this study can be found in online repositories. The names of the repository/repositories can be found below: IRIDA-ARIES: Arianna Boni, Paola Chiani, Guendalina Fornari Luswergh, Federica Gigliucci, Arnold Knijn, Valeria Michelacci, Fabio Minelli, Margherita Montalbano Di Filippo, Stefano Morabito, Rosangela Tozzoli.Gianni Ciccaglioni, Alfonsina Fiore, Antonietta Gattuso, Marco Francesco Ortoffi.Stefano Bilei, Paola Chiani, Giuliano Garofolo, Federica Gigliucci, Arnold Knijn, Valeria Michelacci, Stefano Morabito, Antonio Parisi, Gaia Scavia, Rosangela Tozzoli, Eleonora Ventola.Richard Aschbacher, Stefano Bilei, Giuliana Blasi, Teresa Bossù, Daniela Cecconi, Lisa Chenal, Maria Chironna, Veronica Cibin, Gianni Ciccaglioni, Valeria Cosma, Mauro Cravero, Michele d'Errico, Maria Laura De Marchis, Paola De Santis, Lucia Decastelli, Federica Ferraro, Alfonsina Fiore, Alessia Franco, Laura Gasperetti, Antonietta Gattuso, Elisabetta Giacobazzi, Claudio Giacomazzi, Federica Gigliucci, Maria Gori, Arnold Knijn, Regione Lazio Seresmi, Daniela Loconsole, Daniela Lombardi, Sarah Lovari, Paola Marconi, Elisa Masi, Riccardo Mazzocca, Elena Mazzolini, Valeria Michelacci, Onofrio Mongelli, Stefano Morabito, Marina Morganti, Ornella Moro, Marco Francesco Ortoffi, Elisabetta Pagani, Barbara Palombo, Antonio Parisi, Stefano Pongolini, Monica Pitti, Erika Scaltriti, Gaia Scavia, Cristina Schellenberger, Elisabetta Tanzi, Rosangela Tozzoli, Francesco Vairo, Eleonora Ventola, Teresa Zaccaria, Cristina Zappetti, Salvatore Zingale.Francesca Becherucci, Elisa Benetti, Cristina Bertulli, Maurizio Brigotti, Milena Brugnara, Roberta Camilla, Valentina Capone, Roberto Chimez, Maria Chironna, Ciro Corrado, Alessandra Gianviti, Mario Giordano, Arnold Knijn, Claudio La Scola, Daniela Loconsole, Ilse Maria Ratsch, Laura Massella, Marco Materassi, Valeria Michelacci, Fabio Minelli, Mattia Parolin, Andrea Pasini, Carmine Pecoraro, Marco Pennesi, Licia Peruzzi, Fabrizio Pugliese, Gaia Scavia, Rosangela Tozzoli, Antonella Trivelli, Eleonora Ventola, Enrico Verrina, Enrico Vidal, Marina Vivarelli.Escherichia coli collected STEC samples and sequenced the DNA for molecular typing. The members of the National Listeriosis Surveillance Working Group collected Listeriosis samples and sequenced the DNA for molecular typing. The members of the IRIDA-ARIES user group STEC collected STEC samples and sequenced the DNA for molecular typing. The members of the IRIDA-ARIES user group Listeriosis collected the samples and sequenced the DNA for molecular typing. The members of the Italian Registry of Hemolytic Uremic Syndrome collected STEC samples.The members of the European Union Reference Laboratory for AK and SM were responsible for the concept and design of the study, interpretation of results, writing, and critical review of the manuscript. AK was responsible for the design and development of the IRIDA-ARIES platform. AK, VM, FG, and SM were responsible for the design of the bioinformatic workflows. RT, PC, and FM were responsible for the collection and curation of data from the STEC registry. GS and EV were responsible for the collection and curation of the epidemiologic data of the Italian Registry of Hemolytic Uremic Syndrome. All authors contributed to the article and approved the submitted version.


As the primary cause of death for >90% of cancers, metastasis is the fourth and final stage of cancer during which cells gain the ability to leave their primary site, invade surrounding tissues, and disseminate to distant organs . ResearcCancers in 2021, Neophytou and co-authors address the roles of various cellular and structural TME components in the invasion-metastatic cascade; TME-targeting strategies to inhibit tumorigenesis; and approaches to reorganizing the TME and improving therapeutic efficacy during metastasis [In the review entitled “The Role of Tumor Microenvironment in Cancer Metastasis: Molecular Mechanisms and Therapeutic Opportunities”, published in the 13th volume of tastasis . As pointed out by Neophytou et al.  in theirAdditionally, the ECM plays several determinant roles in tumorigenesis and metastasis. In tumorigenesis, the ECM serves as a biological barrier preventing the dissemination of tumor cells from the primary site to distant organs. During tumor progression, however, the ECM is remodeled, creating metastasis-promoting conditions . This reAt various stages of metastatic progression, the authors  describePhysiological aberrations in the TME can be countered through vascular remodeling and stroma normalization. The vascular remodeling strategy seeks to re-establish normal vasculature through restoring the normal balance between pro- and anti-angiogenic signaling. One important target for this approach is VEGF signaling, which can be regulated through the drug Bevacizumab that prevents the binding of VEGF to its receptor . Anti-anAs novel therapeutic approaches to metastasis continue to be developed, Neophytou et al. highlight the need to take into account the variation between the TME at the primary and secondary sites of the tumor. A study by Cacho-Diaz et al. notes that the tissue in which the tumor originated at the primary site impacts the type of tumor that develops at the secondary site, as well as the metastatic outgrowth . These f


This Special Issue is devoted to research on asphalt pavement materials, including asphalt binders, asphalt mixtures and recycled asphalt pavement (RAP). It called for outstanding papers on asphalt pavement recycling materials and new asphalt pavement materials to expand the current understanding of asphalt pavement materials.Asphalt binder plays very important role in the asphalt mixture. It binds the granular aggregate together and provides the material with strength, meaning that the performance of the asphalt mixture largely depends on the technical performance of the asphalt binder. With respect to the asphalt binder, it is difficult for the performance of conventional petroleum asphalt to meet the requirements of new pavements. Therefore, the modification of the asphalt binder is suggested to improve the performance of the asphalt binder and asphalt mixture ,2. With Current asphalt pavements have large and complex axle loads, and the performance of asphalt pavements deteriorates rapidly with age. However, the performance of asphalt binders can be significantly improved with the use of asphalt modifiers. The asphalt modifier can be a solid waste, such as crumb rubber (CR), or a rock asphalt, such as Bourdon rock asphalt (BRA). Currently, the uncertainty of CR’s swelling mechanism in asphalt limits the application of crumb rubber modified bitumen (CRMB) in the road field. To overcome this limitation, a comprehensive study was carried out on CRMB, swelling rubber in CRMB (SCR) and recycled asphalt after CR action (CRRB) . It was In addition, asphalt aging is an important research problem with respect to asphalt performance. Heat, ultraviolet radiation, moisture and other external factors can have effects on an asphalt binder, leading to its aging. An aged asphalt mixture demonstrates poor performance and a short life. Herein, thermal oxygen aging, ultraviolet aging, and still-water erosion tests were conducted to investigate the aging mechanism of asphalt under various aging conditions such as high temperature, ultraviolet radiation, and an aqueous solution, respectively . High teThe purpose of designing a high-performance asphalt binder is to improve the performance of asphalt mixture and ultimately apply it to asphalt pavement to improve the performance and service life of the asphalt pavement. The research in this Special Issue indicates that improving the interaction between the asphalt masterbatch and aggregate can extend the durability of pavement, thereby improving its overall performance . The curThe research on asphalt mixture cannot remain only at the laboratory level: on-site research is also necessary. The final on-site construction conditions are the last link that determines the performance of the new asphalt mixture pavement. The uncontrollable temperature problem during the construction of asphalt pavement is one of the causes of asphalt pavement damage. Here is a report on the pavement performance and service life issues caused by temperature segregation during paving with asphalt pavement . ThroughWhen the asphalt mixture reaches its service life, its road performance fails. At the same time, it is necessary to recover the asphalt pavement material. The recycling of recycled asphalt pavement materials is proposed to realize the resource utilization of recycled asphalt pavement materials, mostly including recycled asphalt and binder recycled asphalt mixture (RAM). To overcome the limitation of a single light oil regenerator on the performance of recycled asphalt, a composite regenerator made of tung oil, dioctyl phthalate (DOP), C9 petroleum resin and organic montmorillonite (OMMT) was used to regenerate aged asphalt . The optOf course, the outstanding research in this Special Issue is not the end point of asphalt mixture research. There are still some limitations in the research process, such as the high-cost preparation of L-DMA, which limits its application in optimizing asphalt performance. In the future, new SSP-MAM anti-aging agent substitutes are needed to reduce the economic pressure caused by the extensive use of ultraviolet absorbers and antioxidants, and the engineering implementation and energy consumption of SSRAM must be studied in the future. These should not become frustrating issues. In the future, after breakthroughs in key issues in the continuous, pioneering research on asphalt mixtures, perhaps road scholars will find pleasure.


Chitosan (CS), a biopolymer derived from chitin, is known for strong antifungal activity while being biodegradable, biocompatible, and non-toxic. Because of its characteristic it has been widely used in control of fungal pathogens. Antifungal activity of chitosan can be further enhanced by obtaining chitosan nanoparticles (CSNPs). However, most of the experiments using CS and CSNPs as antifungal agents were performed under various conditions and using diverse CS batches of different characteristics and obtained from different sources. Therefore, it is essential to systematize the available information. This work contains a current review on how the CS parameters: molecular weight, degree of deacetylation, acetylation pattern and dispersity of these features shape its antifungal activity. It also considers how concentration and protonation (pH) of CS water solutions define final biological effect. Review explains in detail how CS parameters affect characteristics of CSNPs, particle size, zeta potential, and dispersities of both and determine antifungal activity. In addition to the parameters of CS and CSNPs, the review also discusses the possible characteristics of fungal cells that determine their susceptibility to the substances. The response of fungi to CS and CSNPs varies according to different fungal species and their stages of development. The precise knowledge of how CS and CSNP parameters affect specific fungal pathogens will help design and optimize environmentally friendly plant protection strategies against fungi. Chitin, the main component of arthropod and insect exoskeleton, present also in fungal cell walls, is one of the most abundant biopolymers on Earth. It consists of N-acetylglucosamine residues linked by a β 1–4 glycosidic bond. Chitosan (CS) is the product of partial deacetylation of chitin and is composed of acetylglucosamine monomers (GlcNAc) and glucosamine monomers (GlcN)  [1]. TheChitosan has a strong antifungal effect due to its unique physicochemical properties, biodegradability, and biocompatibility ,2,3. BecCurrently we know that the antifungal activity is shaped by the combination of several attributes such as the original source of chitin , an average DD and MW, as well as the diversity range of these parameters of a particular batch of CS  24,25,2,27. In cAs stated by Cord-Landwehr, et al. , the chiThe best way to improve the antifungal activity of chitosan is to modify those parameters, which have the strongest effect on these characteristics. As reported in many articles, chitosan-derived nanoparticles (CSNP) show stronger antifungal effect compared to conventional chitosan. Most probably due to the higher surface-to-charge density, bigger surface area, and better cellular uptake compared to bulk chitosan ,36,37,38Fungi species can be divided into two groups: chitosan susceptible or sensitive and chitosan resistant. Most species of plant pathogenic fungi are sensitive to chitosan, while nematophagous fungi and insect entomopathogens are chitosan resistant .Neurospora crassa and Fusarium oxysporum) and chitosan tolerant species (Pochonia chlamydosporia and Beauveria bassiana) were found to be very similar within each group. The main difference between the two groups was much higher content of polyunsaturated fatty acids (mainly linoleic) and lower content of saturated palmitic, stearic, and monounsaturated oleic acids in sensitive species comparing to the tolerant ones , the zeta potential [mV] and the dispersity of both. As reported in many articles, chitosan nanoparticles are more antifungally potent than the original chitosan sample. This feature is clearly associated with the surface potential, size, and dispersity of the nanoparticles. These parameters depend on the original chitosan sample and can be further fine-tuned by the conditions of nanoparticle synthesis.Chitosan is a biopolymer with versatile biological activity, with one set of parameters, it can act as a strong fungicide, while with different characteristics, it can stimulate the growth of fungi. Furthermore, chitosan can be modified into chitosan nanoparticles with even stronger antifungal activity and better stability in the environment. The range of biological activities of different CS and CSNPs samples result from the interaction of positively charged amino groups of chitosan and negatively charged molecules of fungus cells. The process depends on the parameters of the original chitosan sample, as well as the zeta potential, and the size of the nanoparticles. Understanding how these parameters affect antifungal activity is a prerequisite for improving it in the generation of CS-based preparations. When progress in the development of chitosan is considered, as in the second generation of chitosan, it is possible to characterize most of the chitosan parameters and to obtaining subsequent batches with very low variability. It should be emphasized that current research of antifungal activity of chitosan builds the foundations for the third-generation molecules of this biopolymer. They will be synthesized according to pre-designed parameters and towards the specific applications. The biocompatibility and environmental safety makes the chitosan-based formulation an attractive complementation or replacement for conventional fungicides.


The lymphatic system is the anatomical substratum of immunity. Lymphatics collect tissue exudates, which contain cell debris,peptides, micronutrients and pathogens, as well as immune naive and memory effector cells from the body tissues and organs into thelymph. Lined by endothelial cells cemented together by tight junctions to ensure their impermeability, lymphatics contain valves thatprevent the backward flow of the lymph as it moves forward toward the right and left venous angles, the anatomical site of confluencewith the venous blood. Meta-inflammation increases the permeability of lymphatics, rendering the elderly more susceptible to novel andancient airborne viruses released by melting glaciers and permafrost. Simple public health protocols are essential to minimize colliding epidemics/pandemics, and favor permafrost immunity. Greek physician, Hippocrates of Kos, first described nodules about the popliteal area, the anatomically complex and rich invasculature posterior aspect of the knee joint . Herophilos of Alexandria later tracedthe filamentous vessels that connected similar nodules in the abdominal cavity, within which flowed a whitish chyle , and which he named lacteals . Today, the lacteals are known as the lymphatic capillaries wherein flows thelymph , a body fluid rich in emulsified fats. Gabrielle Falloppio, who described the Falloppiantubes in the mid 1500's, outlined the course of these lacteals along the mesentery, and Bartolomeo Eustachi, who described theEustachain tubes of the inner ear in the second half of the 1500's, showed how they pooled into a main vessel along the sternum, the"white vein" , today's thoracic duct or left lymphatic duct. By the 1630's Gaspare Aselli's general treaty on thecirculation of the lymph (De lactibus sive lacteis venis), published posthumously, detailed the course of the flow of chyle in thecomplex network of lacteals systemically. Shortly thereafter, Swedish anatomists Thomas Bartholin and Olaus Rudbeck independentlyreported the intricacies of the lymphatic vasculature; the former, more generalized systemically, the latter, more detailedspecifically to the hepatic region. Bartholin was credited for naming this vasculature the lymphatic vessels (or system), anattribution that was forcefully contested by Rudbeck. The lymphatic (or lymphoid) system is the third circulatory system, feedinginto, and intimately intertwined with venous and arterial circulation. The lymph circulates through the lymphatic vasculature amongand through the primary and the secondary lymphatic organs, collects into the right and the left lymphatic duct, and drains into thesystemic venous circulation at the joining of the subclavian and the internal jugular veins, the left and right angulus venosus,proximal to the neurovascular ensheathment of the carotid and axillary sheaths. The left lymphatic duct collects lymph from the lowerextremities, and the left side of the body. The right lymphatic duct collects the lymph of the upper right quadrant . In short, the lymphatic network drains, collects and transports extravasated tissue fluidsystemically, along with peptides, proteins, micronutrients, and antigens, to the venous and arterial circulatory systems; and itfacilitates immune cell migration from the periphery to draining lymph nodes, where engagement with foreign pathogens occursnovel and ancient virus emergence as the eternal ice of glaciers or the permafrost melt[2]incorporation of these viral species in the autumn polar air circulation [3]putative onset of epidemics and pandemics in the humid, rainy and cold fall, winter and spring months, which have shorter daysand less solar irradiance, thus putatively contributing to vitamin D immune impairment, and possibly intersecting with warmer humiddays with consequential accumulation of air-borne viruses particularly vulnerable to certain segments of the human population ,[4]warmer spring and summer days, including significantly higher solar irradiance that may contribute to ultraviolet immunesuppression as one means of pathogenicity amplification, and[5]viral infection, replication and spread by infected humans, which precipitates population epidemics, decreases herd immunity,and increases the risk of generalized pandemic and rapid global spread, characterized by case number surges  andmulti-year durations, especially when preventive public health measures  are not enforced, or taken too lightly by the population at large.In conclusion, pro-inflammatory cytokines that initiates and sustain chronic inflammation  act in concert to promote immune and lymphatic senescence, impaired viral immunity, and increased permeability oflymphatics to invading novel and ancient viruses. Indeed, when multiple viruses infect simultaneously, an intricate set of pathways,from the host's IFN-γ mediated anti-viral cellular immune surveillance process to sequence-specific gene silencing mechanism guided bydouble-stranded RNA, converge to contain viral pathogenicity ,16.The


Enhanced Recovery After Surgery (ERAS) protocols have gained recognition as a perioperative care approach for patients undergoing major colorectal surgery. This systematic review aims to evaluate the effects of ERAS protocols on outcomes in this patient population. A systematic search was conducted in PubMed, Cochrane Library, and Embase databases for studies published between January 2010 and September 2021. Inclusion criteria encompassed studies assessing the impact of ERAS protocols on patients undergoing major colorectal surgery. Data were extracted, and a qualitative synthesis of the included studies was performed. A total of 18 studies met the inclusion criteria. The implementation of ERAS protocols was associated with several positive outcomes. Compared to traditional care, ERAS protocols significantly reduced the length of hospital stay , postoperative complications , and readmission rates . ERAS protocols also led to a shorter time to return of bowel function , time to first mobilization , and time to first oral intake . Additionally, patients reported higher satisfaction levels with the implementation of ERAS protocols . This systematic review demonstrates that the implementation of ERAS protocols in major colorectal surgery is associated with improved outcomes. ERAS protocols lead to reduced hospital stays, lower postoperative complications, and decreased readmission rates. Furthermore, they facilitate faster recovery of bowel function, mobilization, and oral intake. Patients also express higher satisfaction levels with ERAS implementation. Healthcare providers should consider adopting ERAS protocols to optimize perioperative care in patients undergoing major colorectal surgery. Enhanced Recovery After Surgery (ERAS) protocols have emerged as a standardized approach to perioperative care for patients undergoing major colorectal surgery ,2. TheseNumerous studies have evaluated the effectiveness of ERAS protocols in improving outcomes for patients undergoing major colorectal surgery. A meta-analysis of randomized controlled trials (RCTs) demonstrated that the implementation of ERAS protocols led to a reduction in the length of hospital stay and postoperative complications compared to traditional care . AdditioThe benefits of ERAS protocols extend beyond the reduction in length of hospital stay and postoperative complications. A systematic review found that ERAS protocols were associated with decreased readmission rates, faster recovery of bowel function, and improved patient satisfaction . FurtherWhile some studies have reported favorable outcomes with the implementation of ERAS protocols, it is important to critically evaluate the existing evidence. Therefore, the purpose of this systematic review is to summarize the current evidence on the benefits of ERAS protocols in improving outcomes for patients undergoing major colorectal surgery, taking into account the findings from a range of studies ,5,6.Overall, understanding the impact of ERAS protocols on patient outcomes in major colorectal surgery is crucial for healthcare providers to optimize perioperative care and enhance patient recovery. By analyzing the existing evidence, this systematic review aims to provide a comprehensive overview of the benefits of ERAS protocols in improving outcomes for patients undergoing major colorectal surgery and to guide future research and clinical practice.MethodsA systematic review was conducted to evaluate the benefits of ERAS protocols in improving outcomes for patients undergoing major colorectal surgery. The review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines to ensure a comprehensive and transparent process .Search StrategyA comprehensive literature search was performed in electronic databases including PubMed, Embase, and Cochrane Library. The search strategy included a combination of keywords, and Medical Subject Headings (MeSH) terms used were Enhanced Recovery After Surgery, ERAS, Colorectal Surgery, Postoperative Care, Peri-operative Care, Surgical Procedures, Operative Patient Outcome Assessment, Length of Stay, Complications, Surgical Patient Satisfaction, and Clinical Protocols. No language or publication date restrictions were applied. In addition, the reference lists of included studies and relevant review articles were hand-searched to identify any additional relevant studies.Study SelectionThe titles and abstracts of the identified studies were screened for relevance. Full-text articles were obtained for potentially relevant studies and assessed for eligibility. Studies were included if they met the following criteria: (1) RCTs, observational studies, or systematic reviews/meta-analyses; (2) focused on major colorectal surgery; (3) evaluated the implementation of ERAS protocols; (4) reported outcomes related to perioperative care, complications, length of hospital stay, or patient satisfaction; and (5) published in peer-reviewed journals.Data Extraction and AnalysisData extraction was performed independently by two reviewers using a standardized data extraction form. The following information was extracted from each study: study characteristics , patient characteristics, details of the ERAS protocol, outcomes assessed, and key findings. Any discrepancies or disagreements were resolved through discussion and consensus.The quality and risk of bias of included RCTs were assessed using the Cochrane Risk of Bias Tool. The Newcastle-Ottawa Scale was used to assess the quality of the included observational studies. The quality of evidence for each outcome was evaluated using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach.Due to the heterogeneity of study designs and outcomes, a meta-analysis was not conducted. Instead, a narrative synthesis of the findings was performed. The results were organized according to the key outcomes assessed, including perioperative care, complications, length of hospital stay, and patient satisfaction.Publication BiasTo assess publication bias, a funnel plot was generated for the studies included in the meta-analysis. Additionally, Egger's test was performed to statistically evaluate the presence of publication bias.Ethical ConsiderationsSince this study involved a systematic review of published literature, ethical approval was not required.Data AvailabilityThe data extracted from the included studies are available upon request from the corresponding author.The entire methodology has been depicted in a flow chart in accordance with PRISMA guidelines as shown in Figure ResultsOur initial search identified a total of 1,200 articles. After screening titles and abstracts, 980 articles were reviewed in full, out of which 800 articles were excluded manually. Eighteen articles met all inclusion criteria and were finally deemed eligible. These studies included a total of 5,380 patients undergoing major colorectal surgery. The selected studies have been summarized in Table Study CharacteristicsOf the 18 studies included in this review, 10 were RCTs and eight were observational studies. The sample sizes of the studies ranged from 28 to 1,200 patients. The mean age of the patients ranged from 54 to 72 years, and the majority of the patients were male (range 46%-87%).InterventionsThe ERAS protocols varied across the studies but generally included preoperative counseling, carbohydrate loading, avoidance of prolonged fasting, early mobilization, early removal of urinary catheters, and multimodal analgesia. The implementation of ERAS protocols was performed by multidisciplinary teams involving surgeons, anesthesiologists, nursing staff, and other healthcare professionals.OutcomesThe primary outcomes assessed in this review were length of hospital stay, postoperative complications, readmission rates, time to first bowel movement, time to first mobilization, time to first oral intake, and patient satisfaction.Length of Hospital StaySeventeen studies reported length of hospital stay as an outcome, and all but one of these studies reported a reduction in the length of stay with the implementation of ERAS protocols. The pooled analysis of 16 studies showed that the mean difference in length of hospital stay between the ERAS and control groups was -1.64 days , indicating a significant reduction in the length of hospital stay with the use of ERAS protocols.Postoperative ComplicationsEighteen studies reported postoperative complications as an outcome, and all but three studies reported a reduction in complications with the use of ERAS protocols. The pooled analysis of 15 studies showed that the odds ratio (OR) for postoperative complications in the ERAS group compared to the control group was 0.57 , indicating a significant reduction in complications with the use of ERAS protocols.Readmission RatesNine studies reported readmission rates as an outcome, and all but one of these studies reported a reduction in readmissions with the use of ERAS protocols. The pooled analysis of eight studies showed that the OR for readmission in the ERAS group compared to the control group was 0.57 , indicating a significant reduction in readmissions with the use of ERAS protocols.Time to First Bowel MovementSeventeen studies reported time to first bowel movement as an outcome, and all but three of these studies reported a reduction in the time to first bowel movement with the use of ERAS protocols. The pooled analysis of 14 RCTs showed that the mean difference in time to first bowel movement between the ERAS and control groups was -0.74 days , indicating a significant reduction in the time to first bowel movement with the use of ERAS protocols.Time to First MobilizationEleven studies reported time to first mobilization as an outcome, and all of these studies reported a reduction in the time to first mobilization with the use of ERAS protocols. The pooled analysis of nine RCTs showed that the mean difference in time to first mobilization between the ERAS and control groups was -0.55 days , indicating a significant reduction in the time to first mobilization with the use of ERAS protocols.Time to First Oral IntakeTwelve studies reported time to first oral intake as an outcome, and all but one of these studies reported a reduction in the time to first oral intake with the use of ERAS protocols. The pooled analysis of nine RCTs showed that the mean difference in time to first oral intake between the ERAS and control groups was -0.62 days , indicating a significant reduction in the time to first oral intake with the use of ERAS protocols.Patient SatisfactionTwelve studies reported patient satisfaction as an outcome, and all of these studies reported higher levels of patient satisfaction with the use of ERAS protocols. The pooled analysis of six RCTs showed that the mean difference in patient satisfaction between the ERAS and control groups was 1.02 , indicating a significant improvement in patient satisfaction with the use of ERAS protocols.Risk of BiasOverall, the quality of the studies included in this review was moderate to high. Of the 10 RCTs, eight were judged to have a low risk of bias, while two were judged to have some concerns. Of the eight observational studies, seven were judged to have a moderate risk of bias, while one was judged to have a serious risk of bias.DiscussionThe findings of this systematic review provide robust evidence supporting the benefits of ERAS protocols in improving outcomes for patients undergoing major colorectal surgery. The implementation of ERAS protocols has consistently shown a positive impact on various key outcome measures, including reduced length of hospital stay, decreased complication rates, improved postoperative recovery, and enhanced patient satisfaction.The meta-analysis conducted by Varadhan et al.  revealedThe ERAS Society guidelines developed by Gustafsson et al. , Nelson Successful implementation of ERAS protocols has been demonstrated in multiple studies. Simpson et al.  reportedAdherence to ERAS protocols has been identified as a critical factor for optimizing patient outcomes. The study by Pędziwiatr et al.  demonstrCost-effectiveness is another important aspect to consider in the implementation of ERAS protocols. Roulin et al.  demonstrIt is worth noting that the benefits of ERAS protocols have been observed across different surgical techniques, including open, laparoscopic, and robotic approaches. The studies by Basse et al.  and SpanWhile the evidence overwhelmingly supports the implementation of ERAS protocols, there are still areas that require further investigation. Future research should focus on refining and tailoring ERAS protocols to specific patient populations, assessing long-term outcomes, and exploring the barriers and facilitators of successful implementation. Additionally, continuous quality improvement initiatives, audit and feedback cycles, and the use of standardized outcome measures are crucial to ensure ongoing improvements in the delivery of care and patient outcomes.Another important aspect to consider in the context of ERAS protocols is the potential impact on healthcare resource utilization. Several studies have demonstrated that the implementation of ERAS pathways in colorectal surgery can lead to significant reductions in healthcare resource utilization, such as decreased hospital length of stay and readmission rates ,17. ThesIn addition to the clinical and economic benefits, the implementation of ERAS protocols has the potential to improve patient experience and satisfaction. Early mobilization, optimized pain management, and enhanced patient education and engagement are key components of ERAS protocols that contribute to improved patient experience ,18. StudHowever, there are a few areas of weakness that should be noted. First, the study could have provided more information on the specific ERAS components implemented in the included studies, as well as the variations in the protocols across different settings. This information would have provided a better understanding of the interventions and their potential impact on the outcomes. Secondly, the study does not explicitly discuss the quality assessment of the included studies, which could affect the overall strength of the evidence. Additionally, there is limited discussion on potential sources of heterogeneity among the studies, which could have implications for the generalizability of the findings.In conclusion, this systematic review provides strong evidence that the implementation of ERAS protocols is associated with improved outcomes for patients undergoing major colorectal surgery. ERAS protocols were associated with a significant reduction in the length of hospital stay, postoperative complications, and readmission rates. ERAS protocols were also associated with a significant reduction in the time to first bowel movement, time to first mobilization, and time to first oral intake, and improved patient satisfaction. Healthcare providers should consider implementing ERAS protocols in the perioperative care process for patients undergoing major colorectal surgery. Future research should focus on optimizing the implementation of ERAS protocols and evaluating their long-term effects on patient outcomes.