Metabolic reprogramming is one of the main hallmarks of cancer. Numerous studies have shown that the metabolic phenotype of cells within tumors is heterogeneous and distinct from their normal counterparts . FurtherThis Special Issue, “Novel Metabolic Approaches Targeting Cancer Cells”, aims to compile the latest research on tumor cell metabolism and its impact on the development of novel therapeutic targets. The Special Issue contains five articles, two of which are reviews and three of which are original research papers from diverse research groups with remarkable contributions to the metabolism field in cancer.Extracellular vesicles (EVs) are membranous vesicles delimited by a lipid bilayer without replicative capacity. They can range in size from 30 to 1000 nm, are generated by all cells, and have been found in multiple biofluids . They arCarles-Fontana et al. explore the different EV-mediated mitochondrial reprogramming mechanisms supporting cancer survival and progression in their review. Firstly, they detail the main steps in EV biogenesis, release, and cargo in cancer development. Then they summarized a number of EV studies in which the EV-mediated modulation of mitochondrial processes occurred in cancer. Publications about EV cargos carrying various miRNAs derived from cancer cells which were shown to trigger metabolic coupling in the neighboring fibroblasts, or vice versa, are listed. Other EVs can initiate chemotherapy resistance, escape from immune response, or the activation of invasion and the metastasis of tumor cells through the modulation of OXPHOS, the reduction of Bax, and/or an increase in Bcl-2 expression in mitochondria. Cancer-associated fibroblasts have also been shown to transfer mtDNA to breast cancer cells via EV cargos . At the Mátyási et al. investigate how vesicular NME1 and NME2 released by breast cancer cells influence the tumor microenvironment and its metabolism. After overexpressing these two metastasis suppressor genes in human invasive breast carcinoma cells, they found that NME1 was present in medium-sized EVs or microvesicles, whereas NME2 was abundant in both microvesicles and small-sized EVs or exosomes, as well. When human skin-derived fibroblasts were treated with NME1 or NME2 containing EVs, the expression of fatty acid and cholesterol metabolism-related genes decreased significantly in response to NME1 or NME2 containing EV treatment, which showed that these EV-mediated metastasis suppressors directly modify lipid metabolism in fibroblasts in the tumor microenvironment.Oncometabolites are metabolites that accumulate in tumors as a consequence of an alteration in cellular respiration in metabolically rewired cancer cells. This process is considered a hallmark of cancer, driving cancer progression and therapeutic resistance .Low-glycolytic tumors seem to utilize carbon sources other than glucose for bioenergetics and tumor proliferation . CitrateAnother important oncometabolite in the TCA cycle which has gained attention recently is succinate . Casas-BCancer-associated cachexia, defined as involuntary weight loss and breakdown of adipose and muscle tissue, is present in most patients with pancreatic cancer and contributes significantly to mortality . ColemanIn summary, manuscripts published in this Special Issue describe how extracellular vesicles transmit essential factors among cells directly affecting the metabolism of the tumor and its microenvironment, and they can also serve as cargos to deliver various small molecules to target cancer cells. Other papers in this collection highlight the importance of oncometabolites, where the inhibition of their aberrant production can be the potential target of novel metabolic therapies, or where the accumulation of certain metabolites can help to induce the failure of metabolic adaptation in the tumor.


Bifidobacterium longum (BL) NCC3001 has been shown to reduce anxious behavior in preclinical models and feelings of low mood in nonpregnant human adults. During the COVID-19 pandemic, mental health issues increased, and conventionally conducted clinical trials were restricted by social distancing regulations.Perinatal mood disorders such as depression and anxiety are common, with subclinical symptomology manifesting as perinatal mood disturbances being even more prevalent. These could potentially affect breastfeeding practices and infant development. Pregnant and lactating women usually limit their exposure to medications, including those for psychological symptoms. Interestingly, the naturally occurring probiotic This study, Probiotics on Mothers’ Mood and Stress (PROMOTE), aimed to use a decentralized clinical trial design to test whether BL NCC3001 can reduce symptoms of depression, anxiety, and stress over the perinatal period.This double-blind, placebo-controlled, randomized, and 3-parallel-arm study aimed to recruit 180 women to evaluate the efficacy of the probiotic taken either during pregnancy and post partum  or post partum only  in comparison with a placebo control group . Participants consumed the probiotic or matched placebo in a drink once daily. Mood outcomes were measured using the State-Trait Anxiety Inventory and Edinburgh Postnatal Depression Scale questionnaires, captured electronically at baseline (28-32 weeks’ gestation) and during e-study sessions over 5 further time points . Saliva and stool samples were collected longitudinally at home to provide mechanistic insights.In total, 520 women registered their interest on our website, of whom 184 (35.4%) were eligible and randomized. Of these 184 participants, 5 (2.7%) withdrew after randomization, leaving 179 (97.3%) who completed the study. Recruitment occurred between November 7, 2020, and August 20, 2021. Advertising on social media brought in 46.9% (244/520) of the prospective participants, followed by parenting-specific websites . Nationwide recruitment was achieved. Data processing is ongoing, and there are no outcomes to report yet.Multiple converging factors contributed to speedy recruitment and retention of participants despite COVID-19–related restrictions. This decentralized trial design sets a precedent for similar studies, in addition to potentially providing novel evidence on the impact of BL NCC3001 on symptoms of perinatal mood disturbances. This study was ideal for remote conduct: because of the high digital literacy and public trust in digital security in Singapore, the intervention could be self-administered without regular clinical monitoring, and the eligibility criteria and outcomes were measured using electronic questionnaires and self-collected biological samples. This design was particularly suited for a group considered vulnerable—pregnant women—during the challenging times of COVID-19–related social restrictions.ClinicalTrials.gov NCT04685252; https://clinicaltrials.gov/ct2/show/NCT04685252DERR1-10.2196/41751 Subcl changes -5. It is changes , and app changes -8. Furth changes . The con changes -3,8 and  changes ,10. In a changes ,10,11, w changes .Many women opt to avoid taking pharmacological treatments to address feelings of low mood and stress during the perinatal period ,13, ofteLactobacillus rhamnosus HN001, whose primary outcome was infant eczema, showed its potential role in reducing PMD as a secondary outcome [Lactobacillus rhamnosus GG and Bifidobacterium lactis BB12 administered to pregnant women with obesity between 12 and 36 weeks of pregnancy, showed no difference between the intervention and placebo groups in improving prenatal depression or anxiety [A randomized controlled trial of the probiotic  outcome . However anxiety . Other p anxiety ,19. Such anxiety .Bifidobacterium longum (BL) NCC3001 normalized anxiety-like behavior in different preclinical models [Outside of the perinatal setting, the naturally occurring probiotic l models ,22 and rl models . HoweverThis study was designed to test the efficacy of the probiotic BL NCC3001 as a nonpharmacological intervention in reducing PMD. In addition, we are investigating the optimal time window for commencement of probiotic intake to obtain a protective effect: whether there is a need to start the intervention prenatally or whether there would still be a beneficial effect if it is started only after delivery. We hypothesize that daily ingestion of the probiotic BL NCC3001 in the perinatal period will lower time-trend scores of mood and stress as measured by the validated State-Trait Anxiety Inventory (STAI)  and EdinWith the unprecedented advent of the COVID-19 pandemic at the start of 2020, the conduct of many clinical trials was hampered by government-imposed restrictions on face-to-face contact. It was particularly challenging to set up new trials to evaluate interventions unrelated to COVID-19 infections, especially in populations considered vulnerable, such as pregnant women. Clinical trials that could be designed for remote conduct using digital tools were at an advantage . Given tStudy aspects as well as the technical means used for each aspect are presented in RecruitmentDigital advertising Physical posters in public places (public transport and stores)Paper brochure dissemination in mailboxes and partner locationsWord of mouthRegistration of interestSecured online form; participant-initiated self-referralScreening and study sessionsVideo call with screen-sharing capabilitiesSigning of the informed consent formREDCap  electronic databaseDelivery of intervention product and study materials, as well as biological sample collectionTrained and licensed courier serviceSelf-reported questionnairesMedidata Patient Cloud (Medidata Solutions Inc) mobile app in which participants can complete questionnaires Product compliance and all non–self-reported dataiMedidata electronic patient-reported outcome database entry completed by study teamThis trial was set up and conducted in Singapore, where the smartphone penetration rate in the population is 98% and where 42.5% of the population, including 34.3% of women of reproductive age, use social media predominantly to research brands . There iRecruitment in the community at large in Singapore was carried out via self-referral by interested women who heard about the study through the following channels: (1) social media advertising; (2) parenting websites as well as internet forums, platforms, and group chats; (3) physical posters on public transport; (4) brochures in mailboxes and partner locations; and (5) word of mouth. Relevant groups were approached through strategies such as targeted advertising to female users of reproductive age and specific estimated due date groups on various social media and messaging platforms, sending out electronic direct mail messages to members of the parenting websites engaged for the study, and placing physical posters in maternity stores or electronic posters on relevant websites such as the A*STAR Singapore Institute for Clinical Sciences web page.All advertisements included the distinctive, easily recognizable PROMOTE logo. Electronic advertisements contained a convenient web link, and public posters incorporated a QR code to allow potential participants to scan and be directly taken to the website of an official government organization with a secure address to view the eligibility criteria and register their interest. Once they had submitted the forms on the website, the research coordinators would call them to confirm their interest, explain the study, and arrange a screening session via video chat to confirm their eligibility.The study was designed to be conducted completely remotely, with all interaction between study team members and participants taking place via video chat sessions and with collection of biological samples by a licensed courier service.The screening session involved 3 parties  engaged in a video call when the participant was at approximately 27 (range 24-30) weeks’ gestation. If the woman was agreeable to taking part after a detailed explanation of the study, a remote electronic signing of the informed consent form was then undertaken by all 3 parties within the same sitting. The eligibility of the potential participant was confirmed according to the inclusion and exclusion criteria  and immeInclusion criteriaPregnant women aged ≥21 years at recruitmentWilling and able to provide written informed consentGestation of 28 to 32 weeks at randomizationSingleton pregnancy at recruitmentAble to complete questionnaires in EnglishHospital Anxiety and Depression Scale  score ofIntention to breastfeedExclusion criteriaNot willing or unable to comply with the study procedures and requirementsDiagnosed food allergyHas taken probiotic supplements in the 4 weeks before screeningHas received pharmacological treatment for anxiety or depression in the 12 weeks before recruitmentMajor complications during pregnancy Preexisting medical conditions Active participation in another clinical trial or ongoing observational studyThis is a double-blind, placebo-controlled, randomized, and 3-parallel-arm study aimed at assessing time trends in perinatal mood and stress with ingestion of the probiotic BL NCC3001, starting from either the third trimester (28-32 weeks’ gestation) or immediately after birth until 12 weeks post partum, compared with placebo. The 3 arms are (1) prepartum + postpartum probiotic, (2) prepartum placebo + postpartum probiotic, or (3) prepartum + postpartum placebo in a 1:1:1 ratio . ParticiUntil the database lock of the primary outcome, neither the investigator nor the participant nor the site team nor the independent monitor managing the study will know to which investigational product each participant was allocated, with the exception of specified delegated regulatory staff within Nestlé, the product manufacturing site, and the supply and quality managers at the sponsor’s clinical research unit.10 colony-forming units of the probiotic strain BL NCC3001 (8.25%) premixed with a maltodextrin (91.75%) carrier. It is being compared against a placebo of similar sensory characteristics, packaged in a dry powder stick pack containing maltodextrin (84%), yeast extract (12%), pea flour (2%), and cysteine (2%). Both products need to be stored refrigerated and were couriered to participants by designated services trained to maintain the cold chain. The participants were instructed to first dissolve the probiotic in an approved liquid medium  at room temperature or cooler, then consume it within an hour, in the evening with a meal on a daily basis.The daily intervention comprised a dry powder stick pack containing 1.0 × 10The dosage and schedule were identical to those of a previous study , and thiAdherence was assessed by participant recall at each study session. Good adherence was predefined as at least 80% over the study period. Upon the resupply or return of unused products, product consumption was additionally verified.Usual consumption of foods containing probiotics was recorded at baseline (screening session) and again at the end of the study. Participants were requested not to increase the intake of probiotic foods across the duration of the study. In any case, the intake of particular diets, treatments, medications, or concomitant therapies such as probiotic supplements and antibiotics that may interfere with the intervention, as well as medications known to affect mood and stress, during the trial period were captured.The primary outcome is change in time trends of mood and stress scores as measured by the STAI  and EPDSstress index score on a scale ranging from 1 to 5, where higher scores reflect greater stress levels [The secondary outcomes will be derived from validated self-reported questionnaires, including probable depression requiring follow-up (defined as an EPDS score of ≥13) at each time point; change in Hospital Anxiety and Depression Scale score   using video or voice calls accessed via a secure link that maintains data privacy. Participants filled out self-administered questionnaires on their own devices through Medidata Patient Cloud (Medidata Solutions Inc), an electronic patient-reported outcome app that was automatically integrated with the web-based research database . Each study participant was provided with a deidentified personal username and password to access the electronic patient-reported outcome system. For interviewer-administered questionnaires, the study team entered the answers directly into iMedidata. Each participant was assigned a clinical research coordinator for the entire duration of her participation to help build rapport and increase participant retention; regular contact included reminders for upcoming study sessions and the accompanying activities. If a participant became noncommunicative, attempted follow-up was carried out via SMS text messaging or email, and the case was only classed as lost to follow-up or withdrawn after the window period for her final study session had passed.Ethics approval was granted by the A*STAR institutional research board (2020-065), and all women provided signed informed consent. This study is being conducted in line with Good Clinical Practice  and the cold chain to maintain sample integrity. Biological samples are being stored at –80 °C at an appropriately regulated tissue repository until the planned batch analyses at the end of the study. This strategy required comprehensive collection kits to be sent to participants, which included a clear, illustrated, and concise manual, as well as items to alleviate any concerns about needing to store the biological samples in their freezer between the time of collection and courier pickup. These items included laboratory-grade biohazard bags and additional containers to contain the biohazard bags to have multiple degrees of separation between the collected biological sample and the food items in the freezer.Saliva and stool samples were self-collected at participants’ residences and couriered back to the study site by a designated, reliable, and licensed courier trained to respect the Monitoring was carried out periodically by an external, independent monitor. This allowed the evaluation of the trial progress, verification of the accuracy and completeness of electronic case report forms, and resolution of any inconsistencies in the trial records, as well as ensured that all protocol requirements, applicable local laws, International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use  guidelinSafety reporting was conducted in accordance with a preagreed study safety monitoring plan. All adverse and serious adverse events were recorded in the study database.This is an exploratory study of the effect of BL NCC3001 on mood in pregnant and postpartum women; therefore, effect sizes are unknown. However, illustrative power calculations are shown in t test. For each group comparison, the multiplicity coming from comparison at multiple time points is dealt with by performing inference based on the joint distribution of the test statistics from the appropriate set of linear contrasts of the linear mixed model parameters (using the R library multcomp).The trial has 2 primary outcome variables (EPDS and STAI scores) and 2 primary group comparisons (prepartum + postpartum intervention vs placebo control and postpartum-only intervention vs placebo control). Thus, the false positive rate controlled at 5% level is as follows: α / 4 = 0.0125 (for each combination of a primary outcome variable and a group comparison). The time trends for comparing prepartum + postpartum intervention versus control  showed a power of 50% to 70% for observing statistically different time trends when considering recruiting 60 individuals per group, after accounting for an expected dropout rate of 20%, representing 48 participants per group completing the study .With the substantially lower observed dropout rate of 2.7% compared with the initially anticipated 20%, more participants than expected have completed the study, and the power therefore is increased from this initial illustration. Although every effort has been made to obtain complete data sets, the mixed model used for analysis is robust against missing values and valid under missing at random. Outliers will be detected before unblinding, with appropriate action taken considering the medical context. The primary analysis will be an intention-to-treat analysis  with a secondary per-protocol analysis planned. No interim analysis was performed for this trial.Recruitment for the trial commenced on November 7, 2020, and was completed in the following 10 months, with the last participant recruited on August 20, 2021. A total of 520 women expressed interest in the trial through online registration, of whom 205 (39.4%) were agreeable and potentially eligible to participate. Of these 205 participants, 21 (10.2%) withdrew or were found to be ineligible after the screening session, leaving 184 (89.8%) women who were randomized. Of these 184 women, 5 (2.7%) withdrew after randomization, and 179 (97.3%) completed the study . TherefoOf all the recruitment strategies used , advertiThe geodemographic data of Singapore were considered so that resources could be focused on the MRT routes most used by our target population. We first targeted the North-East Line, which services District 19, a catchment area that includes an area with the highest number of residents aged <5 years, an indicator of young families. Almost a quarter  of the prospective participants reported residing in District 19 ,44. The This double-blind, placebo-controlled, and randomized controlled trial will be the first to examine the efficacy of the naturally occurring probiotic BL NCC3001 in alleviating or suppressing symptoms of PMD when taken either from the third trimester or immediately after birth until 12 weeks post partum. It was designed to be conducted entirely remotely in Singapore and was successfully carried out during the height of the COVID-19–related restrictions, using digital technology in all aspects of the conduct of the study while complying fully with research governance and regulations.Although other probiotic interventions have been tested in pregnant women for their impact on PMD with inconsistent results -19, it iThe COVID-19 era has posed many challenges globally. The first challenge concerned the need for social distancing and the associated unpredictable restrictions on in-person activities, including participation in conventional clinical trials. The second challenge concerned the fact that pregnant women are considered a vulnerable group with compromised immunity—in the early stages of the pandemic, very little was known about the detrimental effects of the virus on pregnant women and their unborn child; hence, more precautionary steps were taken to promote their isolation. Therefore, designing a research study that could overcome these hurdles and decrease barriers to participation was crucial to ensure feasibility, continuity, and the eventual successful completion of the study in the face of unpredictable governmental regulations. This study was designed to maintain robust operating procedures that would be resilient to the constant changes in COVID-19–related mandates in Singapore. This was possible because of several reasons, many novel to the local research arena. The remote approach contributed to minimizing inconvenience, reducing exposure risk to COVID-19 infection, and complying with local social distancing regulations. This included all study sessions being conducted virtually (e-visits), which comprised using digital apps to answer self-reported questionnaires and web-based databases that securely synced with these apps, and the intervention products being delivered by courier services, with self-collection of biological samples by participants conducted at their own residences and trained couriers engaged to transport the samples back, with a contactless pickup provided as an option for the participants as well. Despite the apparent success in the recruitment phase, there were some initial challenges before the study could begin. In particular, the ethics board had to be convinced of the reliability and feasibility of remote consenting procedures and approve of the entirely remote conduct of the trial, especially for a population considered vulnerable, such as pregnant women. We also had to design a new secure, acceptable, and legally valid remote e-consenting process to allow consecutive instantaneous signing by a participant, researcher, and witness located at different places, which could be completed rapidly.The widespread ownership and use of personal mobile phone and computerized devices, high availability of internet (both cellular and home internet), trust in digital security systems associated with established local institutions, and greater IT savviness among the target age group , coupledAlthough most of the participants  claimed that social media platforms gave them their first knowledge of the trial, recruitment efforts were strengthened when different avenues of recruitment were used simultaneously. Most notably, when we were targeting recruitment from a geographical perspective, interest from a particular district with a high density of young families peaked when brochures were dropped into residential letter boxes at a similar time to a physical poster advertisement campaign running on the main MRT line servicing that area. We speculate that these different forms of advertisements provided multiple reminders that increased the likelihood of expression of interest. The effectiveness of our nationwide community recruitment strategies is reflected by our data indicating that participants come from all parts of the country.Another aspect integral to the success of the study was the at-point-of-residence self-collection of biological samples. Extensive thought and planning were required to design a biological sample collection workflow that was easy to follow, convenient, and acceptable to participants. Good coordination among the study team, participants, and the licensed courier tasked for pickup was essential. The courier had to follow a robust workflow, maintain the cold chain in tropical Singaporean weather, and demonstrate flexibility in response to constant changes in local COVID-19–related restrictions. Additional benefits that likely attracted participation include increased convenience, attributable in part to no traveling time required to attend sessions. Similarly, this also increased overall time and cost saved for the study team.The actual subject matter of mood disorders also likely resonated more with pregnant women during the COVID-19 pandemic, which helped with study recruitment. Generally increased stress levels across the population have been widely reported worldwide during the pandemic, with pregnant women highlighted as a particularly affected group ,46. We aAs many women try to avoid taking pharmacological treatments during pregnancy and lactation ,13, therMultiple converging factors contributed to the study being able to recruit at speed and retain the required number of participants despite COVID-19–related restrictions. In addition to providing novel evidence for the potential benefit of the probiotic BL NCC3001 in reducing the symptoms of PMD, this entirely remote research study design sets a precedent for similar studies. Such studies could be highly suitable if (1) the intended study population has a high rate of digital device penetration and internet access, (2) systems allow secure data exchange and capture, (3) the intervention can be self-administered without clinical monitoring, and (4) eligibility criteria and outcome measures can be reliably assessed remotely either by digital capture or self-collected biological samples. We have described here such a study that has been successfully conducted in a group considered vulnerable, pregnant women, despite the presence of challenges and barriers presented by the COVID-19 pandemic.


Alloys of magnesium, zinc or iron that do not contain toxic elements are attractive as construction material for biodegradable implants, i.e., the type of implants that harmlessly dissolve away within the human body after they have completed their intended task. The synergistic influence of mechanical stress and corrosive human body fluid can cause sudden and catastrophic fracture of bioimplants due to phenomena such as stress corrosion cracking (SCC) and corrosion fatigue (CF). To date, SCC and CF of implants based on Zn have scarcely been investigated. This article is an overview of the challenges, research needs and way forward in understanding human body-fluid-assisted fractures  of Zn alloys in human body fluid. Avoiding an orthopedic surgical procedure is an extremely attractive proposition in addressing health concerns of ageing populations, which are among the greatest challenges of current times. A surgical procedure can be circumvented when temporary implants  are constructed from alloys that are not only non-toxic and biocompatible but can also harmlessly dissolve within the human body after accomplishing their temporary healing function ,2,3,4,5.Among the structural metals, Mg, Zn and Fe meet the criteria of non-toxicity and biodegradability, which are essential requirements for temporary biodegradable implants. The objective of this article is to present an overview of the comparative merit of zinc (Zn) alloys vis-à-vis magnesium (Mg) and iron (Fe) alloys for non-toxic, biodegradable, temporary implant applications, followed by a thorough discussion on the challenges, research needs and way forward in understanding the human body-fluid-assisted fracture  of Zn alloys in human body fluid.Constructing temporary orthopedic implants from alloys that can harmlessly degrade away in the human body after completing their function has attracted rapidly increasing research and clinical interest. The most significant aspect of such application is to allow the alloy (containing only non-toxic elements) to slowly degrade/dissolve within the human body thus avoiding the second surgery that is usually required to remove the implant.(a)They should not suffer premature failure due to excessive corrosion ,2,3,6;(b)Their elastic modulus (stiffness) should be close to that of human bone in order to avoid “stress shielding” during the healing process ,8. Stres(c)They should possess sufficient mechanical integrity during the entire span of their use as implant ;(d)They must be resistant to sudden brittle fractures that may occur due to the synergy of mechanical stresses and the corrosive environment of human body fluid, i.e., stress corrosion cracking (SCC) and corrosion fatigue (CF) ,8,9,10.The most critical requirements of biodegradable alloys as temporary implants are the non-toxicity of the base metal and its alloying constituents to human physiology, and their ability to degrade away eventually, completely and harmlessly in the human body. The essential requisites of alloys for such applications include:It is essential to ensure the mechanical integrity of the implant device, and that it possesses the desired level of resistance to corrosion and corrosion-assisted fracture for the duration of its temporary use. Therefore, selection of an alloy for temporary implant applications is governed by the constraints of the conflicting requirements of non-toxicity, biocompatibility, adequate mechanical properties and corrosion resistance in human body fluid. Such constraints constitute a narrow window of suitable metals and alloys, and alloys based on Mg, Zn and Fe are the most suitable metallic materials that meet these constraints. However, even for such alloys, some of the requirements may be challenging. For example, precipitation hardening that is commonly employed for strengthening is often deleterious for corrosion and corrosion-assisted premature fracture of Mg alloys, because the strengthening precipitates that are generally highly cathodic to the alloy matrix may cause localized pitting corrosion  and elastic modulus (E = 41–45 GPa) are respectively similar to those of human bones  [2+ ions stimulates tissue growth and healing [2+ ions is harmlessly excreted via urine [Mg possesses the most suitable properties: (a) its low density (–20 GPa) ; (b) Mg Fe is non-toxic, biocompatible and biodegradable in the human body. However, Fe alloys possess a much higher modulus (>>100 GPa) than human bone, creating a risk of “stress shielding”, which leads to weakening of the bone. Another problem with Fe alloys in this specific application is the slow degradation rate.−1) [−1) [E = 80–120 GPa) is not as close to that of bone (E = 3–20 GPa) as that of Mg (E = 41–45 GPa) [Zn corrodes much faster than Fe in the human body, and thereby offers the opportunity to circumvent the issue of the sluggish degradation rate of Fe in temporary implant applications. Both in vitro and in vivo tests suggest that Zn corrodes at considerably slower rates (0.014–0.03 mm y−1) [−1) , which m−1) [−1) , and thu−1) [−1) , which eZn is essential for human physiology. An adult human contains 2–3 g of zinc. Zn is a stimulus for the growth and mineralization of bone tissue, and inhibits bone resorption . The humLi et al.  reviewedThe article by Li et al.  providesAs discussed in the preceding section, Zn alloys meet considerable requisites as a construction material for biodegradable implants. There are a few reports on chemical degradation (corrosion) of Zn alloys in simulated body fluid (SBF). The corrosion and degradation behavior of Zn and its alloys can be determined via in vitro and in vivo studies.3/CO2 on the biodegradation behavior of pure Zn in NaCl solution and SBF, and suggested that the pH of corrosive media that plays an important role in the formation of corrosion products can be mediated using the three buffer systems. Their results indicate that Tris-HCl suppresses the formation of corrosion products and accelerates the corrosion process in both NaCl and SBF, whereas HEPES and NaHCO3/CO2 promote the formation of a passive film and slow down the corrosion rate in NaCl solution, but inhibit the deposition of corrosion products and accelerate the corrosion process in SBF. Furthermore, SBF buffered with Tris-HCl or NaHCO3/CO2 has been proposed as a suitable test medium to assess the in vitro biodegradation behavior of Zn in stent applications. Recently, Wang et al. [The corrosive environment significantly affects the corrosion process and degradation rate of biodegradable Zn alloys, which have been proposed for applications in bone fixation plates and screws, vascular stents, and guided bone regeneration membranes . The serg et al.  confirme2, ZnCO3, Zn3(PO)4 and Ca3(PO)4. The corrosion mechanism and the formation of byproducts are schematically shown in The in vitro corrosion performance of Zn alloys can be evaluated using electrochemical tests including potentiodynamic polarization (PDP) and electrochemical impedance spectroscopy (EIS) in SBF, according to ASTM G59-97. The in vitro degradation rate of Zn alloys can be determined by immersion of the samples in SBF for a defined duration, followed by weight-loss measurements. The commonly used SBF can be either a Hank’s solution, a phosphate-buffered saline (PBS), a Ringer’s saline solution, or a cell culture medium mixed with PBS . The corThe chemical composition and processing procedures  of Zn alloys can play a major role in the corrosion mechanism and nature of the corrosion byproducts . This isThe in vivo corrosion and degradation behaviors of Zn alloys can be assessed using various animal models such as Sprague–Dawley (SD) rats, Wistar rats, mice, beagle dogs, white pigs and rabbits . Zn-alloThere is little reported on the CF and SCC of Zn alloys under physiological conditions, which are critical for bioimplant performance . The simultaneous presence of dynamic loading along with corrosive environment can result in stress corrosion cracking (SCC) and corrosion fatigue (CF), which often occur at stresses considerably below design stresses that would be considered for a non-corrosive environment. The most fundamental and detrimental feature of CF and SCC is that a ductile material that would have undergone considerable elongation before fracture may suffer embrittlement in the presence of a corrosive environment, leading to premature brittle fracture. Because brittle SCC/CF fractures can be sudden, catastrophic and premature, they are the most dangerous forms of corrosion-assisted failures in bioimplants. The dynamic loading in the human body, along with the corrosive physiological environment accentuates the threat of CF and SCC. CF and SCC are of particular concern for devices with sharp contours, such as pins, screws and stents since sharp locations are common stress concentrators, and thus the crack initiation points. There have been several instances of fracture due to SCC of implants for traditional alloys . −5 s−1 and constant load testing while immersed in circulating SBF at 37 °C. The study made a benign claim for the occurrence of SCC on the basis of a comparison of stress-strain data generated for the alloy in SBF and air, but did not make a conclusive inference on SCC. In general, it is critical to understand the role of strain rate in the susceptibility of an alloy to SCC. The strain rate employed in SSRT for investigating SCC is often in the range of 10−6–10−7 s−1, as established for various alloy systems such as Mg alloys in SBF [−5 s−1 that was employed for SSRT in the recent study on SCC of Zn-0.8Li [−7–3.1 × 10−7 s−1 and suffered SCC, as confirmed by representative fractographs revealing transgranular crack propagation and crack branching in SBF  , a Zn-0.s in SBF ,38,39,40s in SBF ,42. Therg in SBF . As show−7 s−1, but no SCC was observed when the strain rates were either higher or lower than 3.1 × 10−7 s−1. These findings have been corroborated by the fractographic features  often suffer extensive pitting due to secondary precipitates that are commonly developed in alloys for the purpose of strengthening . Most se(a)Maximize solid-solution strengthening by selecting alloying elements that have high solubility and also meet the non-toxicity criteria for body implant applications;(b)Employ suitable thermomechanical treatments to control the grain size as a complementary strengthening measure.In light of the restrictions described above, suppression of precipitation in Zn alloys may be a challenging task. However, there is great value in developing Zn-alloy implants that possess adequate resistance to human body-fluid-assisted fractures. Therefore, there may be a need to:2Mg5Zn5 [2Ca), for manipulating pitting susceptibility. A similar strategy may be pursued for Zn alloys. However, it is noted that the strain-hardening characteristics of Zn alloys during extrusion will be considerably different, and the processing parameters will thus need to be thoroughly optimized.For example, among the alloying elements that meet the non-toxicity criterion, manganese (Mn) possesses significant solubility in Zn , and hen2Mg5Zn5 ) to anodA proper simulation of the chemical composition of the body fluid and ensuring that mechanical factors have been appropriately considered is critical for generating accurate data for corrosion and corrosion-assisted fracture of bioimplants.As elaborated earlier, in vitro corrosion studies on biodegradable metals in simulated body fluids (SBFs) have employed many types of pseudo-physiological solutions that mimic the composition of body fluids ,52,53; f2. As shown in 2 bubbling through SBF (instead of HEPES) to control pH was employed in recent studies on CF (and SCC) of biodegradable alloys [Another critical aspect concerns the appropriate means for controlling the pH of the simulated body fluid (SBF), which is commonly achieved using buffering chemicals, such as 2-(4-(2-hydroxyethyl)-1-piperazinyl) ethanesulfonic acid (HEPES)). However, HEPES can interfere with the main corrosion reaction . The acte alloys ,58.Metallurgical characteristics such as grain size and texture, which develop as a result of thermomechanical processing such as extrusion, have been found to influence the corrosion of magnesium alloys , and henLittle has been reported on the influence of physiological molecules such as proteins and glucose content of blood plasma in corrosion and corrosion-assisted cracking  of Zn alloys. In particular, there are no reported studies on SCC and CF of such alloys in SBF that considered both inorganic and organic constituents of body fluid. A study that investigated SCC and CF of a Mg alloy in SBF with and without the most abundant organic constituent of body fluid, i.e., bovine serum albumin (BSA), found BSA addition to significantly increase susceptibilities to SCC but, intriguingly, BSA addition retarded corrosion fatigue . Such stKISCC). The scientific/mechanistic basis for the CNT technique to generate accurate KISCC data has also been validated. The CNT testing and associated specimen fabrication is simpler and less expensive  [Another aspect that needs particular attention is that the devices often consist of both sharp and smooth contours. While SCC and CF can also initiate on smooth metallic surfaces, sharp contours are the most susceptible locations for SCC and CF initiation. Therefore, the most common SCC and CF data used for design purposes are generated using specimens with pre-existing sharp cracks/notches. Smooth specimens are tested for establishing the susceptibility of a metallic material to cracking, whereas tests using specimens with pre-existing sharp cracks/notches establish the susceptibility of an existing crack to propagate under the influence of a corrosive environment. Therefore, it is essential to test pre-cracked specimens of identified Zn alloys, and such data are currently non-existent for Zn alloys in body fluid. Circumferential notch tensile (CNT) testing  is a costesting) , which tBecause corrosive body-fluid-assisted premature fracture is a concern for bioimplants, it is necessary to understand and address such fracture before implants of such materials are put into actual service. This article briefly discusses the suitability of Zn, Mg, and Fe alloys for their use as biodegradable implant materials, and critically assesses recent investigations on stress corrosion cracking (SCC) of Zn alloys as potential bioabsorbable implant materials. One of the critical aspects in this article is to illustrate the need of comprehensive investigations of SCC and corrosion fatigue (CF) of Zn alloys in physiological environments. This article identifies in particular the critical knowledge gaps in SCC and CF investigations on Zn alloys in appropriately simulated mechanical loading and human body-fluid conditions.


Scoliosis is a three-dimensional structural deformity of the spine; more than 80% of scoliosis has no specific pathogenesis but is understood to be closely related to genetic, hormonal, and environmental factors. In recent years, the epigenetic alterations observed in scoliosis have been analyzed in numerous studies to determine the pathogenesis and progression of this condition, however, there is currently no comprehensive review of the epigenetic factors to date. We searched PubMed, Embase, and Web of Science databases for relative studies without language and date restrictions in March 2023. Twenty-five studies were included in this review and analyzed from the four main aspects of epigenetic alteration: DNA methylation, non-coding RNAs, histone modifications, and chromatin remodeling. The relationship between DNA methylation, non-coding RNAs, and scoliosis was considerably reported in the literature, and the corresponding related signaling pathways and novel biomarkers observed in scoliosis provide insights into innovative prevention and treatment strategies. However, the role of histone modifications is rarely reported in scoliosis, and few studies have investigated the relationship between scoliosis and chromatin remodeling. Therefore, these related fields need to be further explored to elucidate the overall effects of epigenetics in scoliosis. Scoliosis is a generic term that describes a three-dimensional structural deformity of the spine , primariEpigenetics is defined as variations in gene expression that are not caused by DNA sequence alterations and are heritable in mitosis and meiosis . Key epiIn this review, the terms “scoliosis,” “epigenetics,” “DNA methylation,” “histone modification,” “chromatin remodeling,” and “non-coding RNA” were used as keywords to find relative articles in PubMed, Embase, and Web of Science databases. This search was conducted until March 2023. The inclusion criteria were as follows: 1) clinical or basic research relevant to the above keywords; 2) papers published in journals with no language restrictions. The exclusion criteria were as follows: i) papers without full-text availability; ii) research with low relevance to the topic; iii) research with an unclear description of the original text or a relatively insufficient evidence-based rating. A total of 487 relevant papers were searched, and 25 papers were finally included in this review according to the selection criteria. Ten articles were related to DNA methylation, 13 related to noncoding RNA, 2 articles introduced histone modification, and we did not find article focused on the relationship between chromatin remodeling and scoliosis. The source of samples in each article were summarized in DNA methylation within the human genome typically occurs on the cytosine of the CpG dinucleotide, which is often located at the starting point of gene transcription . Under tCOMP gene from patients with AIS and healthy controls  is an extracellular matrix component essential for cartilage growth. Some studies have demonstrated that the secretion of COMP is significantly lower in patients with AIS than in normal controls . Mao et controls . The levPITX1 gene was hypermethylated, and the expression levels of downstream products were markedly reduced in patients with AIS compared to those in controls  promoter 0N in the paravertebral muscle was significantly higher on the concave side of the patient than that on the convex side. Corresponding correlation analysis revealed that ESR2 promoter 0N methylation variation was strongly associated with the development of AIS but was not significantly associated with the degree of curvature. This study was the first to elucidate a possible mechanism for the development of scoliosis from the perspective of aberrant local tissue DNA methylation. Janusz et al. (ESR1) gene. ESR1 expression was determined to correlate with the methylation of CpGs in the T-DMR2 of the concave paravertebral muscle. The methylation of the four CpG islands in the T-DMR2 of the concave paravertebral muscle was markedly low in patients with less severe scoliosis (Cobb’s angle <70°). These results establish a correlation between aberrant local tissue methylation at key loci and the progression of scoliosis.Alternatively, other studies focused on DNA methylation alterations that are specific to skeletal muscle tissue around the spine of patients with scoliosis. In 2020, Chmielewska et al. examined DNA methylation levels in deep paravertebral muscle samples taken from the convex and concave sides of patients with AIS . The metz et al. assessed RARA gene on chromosome 17), cg08826461 (corresponding to an unknown gene on chromosome 2), cg12922161 (within the LOC150622 gene on chromosome 2), and cg16382077 (corresponding to an unknown gene on chromosome 7) were the four most highly correlated DMRs observed in this study. Most of the corresponding gene products were enriched in the Wnt signaling pathway, which is associated with the production of neuropeptide Y (NPY). The Wnt signaling pathway has been previously reported to be important for bone formation and remodeling  at a range of loci often differ between patients with scoliosis and normal controls; therefore, in addition to aberrant methylation at key loci, attention has been provided to the impact of aberrant methylation sites across the genome. Since monozygotic (MZ) twins are genotypically identical but phenotypically different , the cormodeling ; additiomodeling . Aberranmodeling .In recent years, an increasing number of studies have demonstrated that non-coding RNAs play an essential role in the regulation of gene expression in the development of various diseases . The mosLiu et al. revealed a series of 546 mRNAs with 139 lncRNAs differentially expressed in AIS children and controls . In partin vitro cytological assays; moreover, this fraction of knocked-down cells failed to effectively form bone. This study further established that in normal humans, lncAIS interacted with NF90 to promote HOXD8 mRNA stability, thereby enhancing RUNX2 transcription in BM-MSCs and safeguarding the osteogenic differentiation of normal BM-MSCs. Within the AIS population, a significantly low expression of lncAIS causes impaired osteogenesis of the differentiated MSCs, resulting in the development of scoliosis  of patients with AIS and normal controls . Specificoliosis .H19 gene, was observed to be significantly under-expressed in the concave paravertebral muscle tissue of patients with AIS, implying that this lncRNA plays an important role in the regenerative differentiation of skeletal muscle   of Foxo4, thereby downregulating Foxo4 levels. Luciferase and siRNA assays revealed that SULT1C2A downregulates rno-miR-466c-5p expression through the PI3K-ATK signaling pathway, thereby increasing Foxo4 expression. Foxo4 is a forkhead box (Fox) transcription factor that participates in the development of cartilage and skeletal muscle during embryonic development  . SULT1C2elopment . TherefomiRNA is a small type of RNA that can bind to mRNA to promote its expression or breakdown. Overall, miRNA contributes to the development of scoliosis through the action of signaling pathways. In 2017, Jiang Heng et al. selected muscle tissue from the convex and concave sides of the paravertebral vertebrae of three female patients with AIS ; correspin vitro. miR-145 expression was positively correlated with the expression of CTNNB1, which encodes β-catenin; further, continued activation of β-catenin may impair osteoblast function in patients with AIS. After knocking down the miR-145 gene in AIS osteoblasts, miR-145 expression was downregulated, the formation of an active β-catenin/Tcf4 complex was significantly inhibited, and normal expression of osteoblast markers, including osteoprotegerin (OPG), transmembrane glycoprotein E11, and sclerostin secretion (SOST), was restored. Additional specific disorders can accompany scoliosis. In 2018, Seco-Cervera et al. analyzed miRNA differences between patients with Friedreich’s ataxia (FRDA) and controls , five patients with severe AIS (Cobb angle >50°), and five healthy controls and sequenced their serum samples for miRNA . Correspcontrols ; in thisSome studies have also used animal models to explore the role of miRNAs in scoliosis. Ishiwata et al. determined that miR-224-5p expression was greatly upregulated in the lumbar spine of a CS model (Ishibashi rats) compared to that in normal 42-day-old rats . FurtherIn recent years, the possibility of utilizing specific circRNAs in the serum as diagnostic biomarkers for scoliosis-related diseases has been reported. García-Giménez et al. reported differences in the distribution of circRNA abundance between patients with AIS and normal controls . They foSome studies have also focused on the overall variation of the three types of non-coding RNAs in patients with scoliosis. In 2018, Chen et al. sequenced non-coding RNAs with mRNAs in VAD-CS rat embryos . A totalIn 2020, Li et al. searched PubMed, EMBASE, and GEO databases for literature that compared differences in gene, miRNA, and lncRNA expression between patients with AIS and normal control MSCs ; a compuHistone modification such as methylation and acetylation is an epigenetic modification that affects transcriptional activity . Mao et In 2019, Li et al. performed histological and genetic testing of articular cartilage from 11 patients with IS compared to 10 matched controls . Light mChromosomes consist of nucleosomal units of DNA bound to histones, which enables the genome to be highly condensed . ChromatEpigenetic regulation plays an essential role in the pathogenesis of scoliosis. Consequently, recent studies have explored the relationship between scoliosis and epigenetics, with primary focus on two mechanisms: DNA methylation and non-coding RNA regulation. These mechanisms have specifically been evaluated by analyzing the changes in the expression of key products and their corresponding influence on signaling pathways, leading to a deeper understanding of the pathogenesis of scoliosis. However, the small number of studies that can be cross-examined may affect the corresponding depth of understanding within this review. Most studies analyzing epigenetics in scoliosis have not yet adopted methods such as downstream protein analysis to validate their proposed theories of pathway effects; therefore, there remains the requirement for further studies to deepen the understanding of DNA methylation and non-coding RNA regulation in scoliosis by testing the validity of current studies and further evaluating the mechanisms involved in this pathogenesis. Further, there are few studies on histone modification and chromatin remodeling, and the effects of chromatin remodeling on scoliosis are yet to be elucidated. Because the process of chromatin remodeling can be affected by a range of cellular behaviors, such as replication, transcription, and repair, the corresponding research is less reliable; therefore, developing an understanding of chromosome remodeling patterns specific to patients with scoliosis will likely be challenging. To conclude, epigenetic research remains an emerging field in scoliosis, and further studies are required to improve our current understanding of the pathogenesis of scoliosis and provide new insights into the future clinical diagnosis and treatment of this disorder.


Thrombotic thrombocytopenic purpura (TTP) is a fatal disease in which platelet-rich microthrombi cause end-organ ischemia and damage. TTP is caused by markedly reduced ADAMTS13  activity. ADAMTS13 autoantibodies (autoAbs) are the major cause of immune TTP (iTTP), determining ADAMTS13 deficiency. The pathophysiology of such autoAbs as well as their prognostic role are continuous objects of scientific studies in iTTP fields. This review aims to provide clinicians with the basic information and updates on autoAbs’ structure and function, how they are typically detected in the laboratory and their prognostic implications. This information could be useful in clinical practice and contribute to future research implementations on this specific topic. Thrombotic thrombocytopenic purpura (TTP) is a rare hematological disorder caused by a deficiency in the enzymatic function of a member of the disintegrin and metalloprotease with thrombospondin-type motifs family, an enzyme called ADAMTS13, which is synthesized primarily in the liver and, in limited quantities, by vascular endothelial cells, megakaryocytes, platelets, glomerular podocytes and glial cells. ADAMTS13 binds soluble von Willebrand Factor (VWF) and interacts with endothelium-anchored Ultra-Large VWF Multimers (ULVWFMs), resulting in the cleavage of ULVWFM strings or bundles to regulate their interaction with platelets, thus preventing the formation of blood clots in normal circulation. In patients with immune TTP (iTTP), ADAMTS13 activity is significantly reduced due to the binding of anti-ADAMTS13 autoantibodies (autoAbs) to the metalloprotease. Consequently, ULVWFMs remain uncleaved in circulation, forming platelet-rich thrombi in the microvessels under conditions of high shear stress [As in other autoimmune disorders, iTTP is characterized by a loss of tolerance resulting in a shift to autoimmunity . AntigenThe autoimmune response against ADAMTS13 is polyclonal and heterogeneous ,7. A stuSeveral scientific groups are working to better understand which epitope/epitopes of ADAMTS13 these autoAbs recognize and bind to. Several authors have demonstrated that a major binding site for autoAbs in iTTP is the cysteine-rich/Spacer (CS) domain ,15,16,17Increasing knowledge on autoAb immunoprofiles might support the improvement of targeted therapies for better iTTP patient management. For example, if a patient has only anti-S autoantibodies, an rADAMTS13 variant, mutated in the S domain, could be used to escape the binding of anti-S autoAbs ,24. How do ADAMST13 autoAbs work? ADAMTS13 deficiency may manifest with reduced activity and/or a reduction in circulating antigen depending on the autoAbs mechanism. Thus, the autoAbs response to ADAMTS13 includes neutralizing and/or non-neutralizing antibodies. Neutralizing autoAbs block the proteolytic activity of ADAMTS13 towards VWF, primarily reducing the enzymatic function of the molecule, while non-neutralizing autoAbs may contribute to increasing ADAMTS13 clearance or interfere with ADAMTS13 interaction with cells or other plasma proteins. As previously reported, anti-CS domain autoAbs mainly inhibit the function of ADAMTS13 by targeting the S domain, which plays a key role in VWF binding . Thus, tRecently, Underwood and colleagues demonstrated the enhanced rate of ADAMTS13 clearance and how this appears to be the major mechanism of reduced ADAMTS13 activity during plasma exchange (PEX) . The autIn summary, the development of ADAMTS13 autoAbs is the result of an immune imbalance involving B cells, cTfh cells and plasmablasts, leading to a polyclonal autoimmune response. Most ADAMTS13 autoAbs belong to the IgG family and recognize the CS domain of the ADAMTS13 molecule. One matter of debate is the main function of autoAbs, neutralizing or non-neutralizing, in the enhancement of ADAMTS13 clearance. Increasing knowledge about the structure, function and specific epitope recognition of ADAMTS13 autoAbs not only helps to better understand iTTP pathophysiology but, as seen here, could have important clinical-therapeutic implications in the future, especially with the advent of new molecules.A test of ADAMTS13 activity is required to confirm TTP diagnosis. Then, in order to differentiate iTTP from congenital TTP (cTTP), the identification of ADAMTS13 autoAbs is mandatory. Diagnostic samples for ADAMTS13 activity and autoAbs testing should be collected prior to treatment . Severe Bethesda assays are used to detect and titer neutralizing autoAbs. Test plasma is heat-treated to inactivate any ADAMTS13 still present, leaving autoAbs in the plasma intact. One volume of heat-treated plasma is added to one volume of normal pooled plasma (NPP), the source of ADAMTS13 in the assay. A separate control mixture is prepared, comprising equal volumes of NPP and buffer; in this way, both tubes begin the procedure with identical levels of ADAMTS13. Then, both tubes are incubated for 30 to 120 min, depending on the protocol, to permit the formation of antigen–antibody complexes. The ADAMTS13 activity of the test and control mixtures is then measured in a functional assay and the residual ADAMTS13 activity of the test sample is calculated as a percentage of that of the control sample subjected to identical incubation conditions. One Bethesda unit (BU) is an inhibitor titer that decreases the residual activity to 50% of the expected value. Performing a dilution series allows for titer determination. Rather than wait for the result on just the 1 + 1 dilution, it is common to perform the assay on a range of dilutions in the first instance, then correcting for the dilution factor . Some auThe tests commonly referred to as ADAMTS13 autoAbs tests utilize enzyme-linked immunosorbent assay (ELISA) . ELISA iELISA-positive/Bethesda-negative results have been described in recovered iTTP patients who showed both ELISA- and Bethesda-positive autoAbs results at the time of acute iTTP diagnosis, confirming that inhibition is not necessarily the primary effect of some ADAMTS13 autoAbs. Thus, using only the Bethesda assay will lead to the under-detection of ADAMTS13 autoAbs  and geneTogether with ADAMTS13 activity and other potential prognostic markers, several authors have evaluated whether and what role ADAMTS13 autoAbs could have in a prognostic sense, considering both the impact on severity and mortality, and on the recurrence risk. Ferrari et al. observed that patients included in their study were less likely to survive their first iTTP event if they had IgG1 and very low or undetectable IgG4 levels plus higher titers of other classes of ADAMTS13 autoAbs (particularly IgA), suggesting that an immune response characterized by high levels of IgG4 could, at least partially, predict a more treatable form of iTTP . The autWhile the significance of reduced ADAMTS13 activity during remission is more consolidated, the predictive value of disease recurrence determined by the presence of ADAMTS13 autoAbs, their inhibitory activity and Ig classes and subclasses is still controversial. In different studies, ADAMTS13 autoAbs during remission emerged as one of the possible risk factors associated with an increased risk of iTTP relapse, together with young age, race, a previous relapse of iTTP and severely deficient ADAMTS13 activity in remission ,57,58. PSumming up, together with ADAMTS13 activity and other clinical and laboratory factors, the role of ADAMTS13 autoAbs has been and is currently an interesting object of research. Even if global agreement between authors has not been reached, ADAMTS13 autoAbs are emerging as possible markers of a more severe acute form of iTTP, associated with a longer time to platelet count recovery, a higher number of PEX sessions, mortality and burden of care. Such conclusions could also be used to optimize the selection of those patients needing early immunosuppressive treatment intensification with RTX or, on the other hand, those with good chance to achieve clinical response with only steroid therapy. Furthermore, ADAMTS13 autoAbs together with ADAMTS13 activity may also represent a useful tool in the remission phase, able to identify patients with higher recurrence risk and with possible indication for pre-emptive treatment.ADAMTS13 autoAbs represent only one of the interesting and not fully understood aspects of iTTP. Much progress in understanding the pathophysiological mechanisms has been made. Interest in such mechanisms is growing given the important clinical implications they may have with the advent of new therapeutic molecules. Their prognostic value is another important aspect. Although a unanimous consensus cannot be drawn by published studies, ADAMTS13 autoAbs appear to play a role both in the identification of more severe forms of acute iTTP and as a marker of recurrence risk during the remission phase. Thus, the refinement of laboratory methods aimed at their detection and characterization as well as large data collections will represent a cornerstone in the world of iTTP for the coming years.


Tellurium is a super-trace metalloid on Earth. Owing to its excellent physical and chemical properties, it is used in industries such as metallurgy and manufacturing, particularly of semiconductors and – more recently – solar panels. As the global demand for tellurium rises, environmental issues surrounding tellurium have recently aroused concern due to its high toxicity. The amount of tellurium released to the environment is increasing, and microorganisms play an important role in the biogeochemical cycling of environmental tellurium. This review focuses on novel developments on tellurium transformations driven by microbes and includes the following sections: (1) history and applications of tellurium; (2) toxicity of tellurium; (3) microbial detoxification mechanisms against soluble tellurium anions including uptake, efflux and methods of reduction, and reduced ability to cope with oxidation stress or repair damaged DNA; and (4) the characteristics and applications of tellurium nanoparticles (TeNPs) produced by microbes. This review raises the awareness of microorganisms in tellurium biogeochemical cycling and the growing applications for microbial tellurium nanoparticles. Tellurium (Te) was first discovered in 1783 from the tellurium-bearing gold mining area of the Metaliferi Mountains, in modern-day Romania Emsley . Its cruThe usage of tellurium may have also increased dramatically if tellurium compounds had been used to prevent engine-knocking of combustion engines – despite their efficacy, their smell meant that insidious tetraethyl lead was used instead, to environmentally detrimental effect  Anderson , but a m32−) and tellurate (TeO42−), in natural waters and weathered surface environment geological samples  is less toxic than both  taken orally was absorbed by the gut; tellurium was primarily stored in the kidneys, but it accumulates in the liver, spleen, heart, lungs, brain, and bones  interference of thiols (redox enzymes); 2) replacement of Se and S in proteins; (3) damage of cell membrane structure; and (4) increased oxidative stress  activity, soil bacterial count and wheat root length  of tellurite  is a typical efflux protein that could eliminate tellurite out of cells  oxide respiring bacteria around black smokers is known to remove tellurium  oxyanions as terminal electron acceptors through anaerobic respiration 2Te, DMTe], which is less toxic than tellurite, highly volatile and has a characteristic garlic odour 2Te2, DMDTe] and the mixed species dimethyltellurenyl sulfide   in Tellurite or tellurate react with reduced thiols or reductases after entering the cell, leading to production of reactive oxygen species (ROS) , alkyl hydroperoxide reductase (AhpCF), glutathione reductase (GorA), glutaredoxin A (GrxA), and thioredoxin C (TrxC) to cope with oxidative stress  and endonuclease IV to deal with the toxicity of ROS produced by tellurite  reduction of ROS or (2) repair DNA damage caused by ROS to cope with the increasing ROS. For instance, the production of ROS led to the up-regulation of global regulatory factors such as OxyR and SoxRX in S. barnesii and Bacillus selenitireducens  and improved the survival rate of infected animals, while also showing a reasonable level of safety in terms of liver and kidney function. Bio-TeNPs are expected to be an alternative to traditional antibiotics and chemical fungicides used in antibacterial coatings of medical devices  Biod et al.  tested BIn recent years, with the increasing demand for tellurium, concerns about environmental pollution and toxicity associated with tellurium have gained widespread attention. Microbes play an important role in the tellurium biogeochemical cycle. Several different mechanisms for tellurium detoxification have been identified in microorganisms. Among the detoxification mechanisms, the conversion of highly toxic tellurate and tellurite oxyanions into lower-toxicity elemental tellurium or methylated organotellurium compounds through microbial processes has been demonstrated as an environmentally friendly, feasible, and promising approach to deal with tellurite and tellurate contamination. Moreover, some microorganisms could reduce tellurite or tellurate to elemental tellurium and form Bio-TeNPs. Bio-TeNPs show great potential applications in fields including biomedicine, optoelectronics and environmental remediation. Future research is expected to focus on the following aspects: (1) tellurium pollution monitoring in the environment, (2) migration and transformation of tellurium in the natural environment, (3) tellurite and tellurate reduction mechanisms under anaerobic conditions, (4) high efficiency tellurite and tellurate reductases, (5) identification of tellurate methylase, (6) Bio-TeNPs formation and secretion mechanism, (7) the toxicity evolution of Bio-TeNPs, (8) more application directions of Bio-TeNPs.


Carcinogenesis, the process by which normal cells transform into cancer cells, is complex and multifaceted. Understanding the molecular mechanisms underlying this multitude of diseases is crucial for the development of effective diagnostic tools and targeted therapies. Transcription factors (TFs) play a central role in regulating gene expression and have emerged as key players in the development and progression of various cancers. In simple terms, a transcription factor  is a specialized protein that binds to DNA and governs the rate of transcription of genetic information from DNA to messenger RNA ,2. BecauCancers, five articles highlight the important role of certain transcription factors in carcinogenesis.In the context of carcinogenesis, the aberrant activation or inactivation of TFs can disrupt the delicate balance of gene expression, leading to uncontrolled cell growth and the formation of tumors. Recent research has underscored the significance of distinct transcription factors in various cancer types, shedding light on their roles and potential therapeutic implications. In this Special Issue of Two of the articles, one a review  and the As the authors indicate, the TIME is a highly complex system comprising both innate and adaptive immune cells that play a central role in the development and progression of cancer, including HCC . Based oOverall, this is a comprehensive and informative review focusing on HCC and the tumor immune microenvironment. Within this complex microenvironment, the SOX transcription factors assume a crucial role, orchestrating gene expression and influencing a myriad of processes both within the tumor and its surrounding tissue. Among these factors, SOX transcription factors are emerging as key regulators of the tumor immune microenvironment, wielding their influence over immune cells, stromal cells, and the overall tumor microenvironmental landscape. This newfound understanding positions SOX transcription factors as promising targets for cancer research and potential therapeutic interventions aimed at enhancing the efficacy of cancer treatments .SOX family genes encode a group of proteins that share a similar DNA-binding high-mobility-group (HMG) domain [SOX1 is hypermethylated in various cancers [The research article focusing on SOX published in this Special Issue delves into the enigmatic role of SOX1 in lung cancer, unearthing its robust potential as a tumor suppressor through its effective blocking of the hairy and enhancer of split 1 (HES1) . The HES) domain  and are ) domain ,13,14,15) domain ,17,18. T cancers ,20,21, i cancers .SOX1 in lung cancer does indeed contribute to SOX1 silencing or downregulation. Furthermore, the restoration of expression of SOX1 in cells or mice suppresses lung cancer growth and invasion. Additional experiments exploring possible mechanisms for these effects show that SOX1 directly targets the HES1 promoter to reduce its expression. The downregulation of HES1 was confirmed to contribute to the effectiveness of SOX1 suppressive effects against lung cancer cell growth. In conclusion, researchers show that SOX1 performs its tumor suppressor function by directly binding to the HES1 promoter and thus SOX1 may be a critical tumor suppressor gene in lung cancer. These findings [In the current study , they cofindings  not onlyA second review article  in this This particular review concentrates on categorizing and listing the transcription factors involved in the EMT based on their various DNA-binding domains (DBDs). These domains include TFs containing DNA-binding basic domains, zinc-coordination DNA-binding domains, helix-turn-helix binding domains, other all-alpha-helical DNA-binding domains, immunoglobulin fold DNA-binding domains, and beta-hairpin exposed by an alpha/beta-scaffold DNA binding domains.An overview of the epigenetic regulation pathways of TFs involved in the EMT is also presented followed by the therapeutic implications and challenges of targeting EMT-TFs in cancer. Currently no drugs have been approved by the FDA because of the complexity of the EMT. Thus, understanding the roles of various TFs is vital for developing targeted therapies and identifying specific biomarkers for personalized treatments. Overall, this review provides a comprehensive systematic classification of the various types of TFs involved in the EMT process based on their DNA-binding domain structure .Two of the five articles published in this Special Issue are associated with cancers that are difficult to treat due to their complex heterogeneity or their resistance to therapy.The additional compelling signaling pathways include those mediated through the primary cilium (PC), a long, thick organelle that protrudes from the tip or apex of almost all cell types, particularly epithelial cells. The primary signaling pathway associated with the PC is probably the hedgehog (Hh) pathway. The exact role of primary cilia in transducing Hh signaling is complex and context-dependent because cilia can act as both positive and negative mediators of Hh signaling. Deregulation of Hh signaling has an important role in the development and progression of cancer, especially MPM. In this Special Issue, researchers investigated the presence of the PC in malignant pleural mesothelioma (MPM) , a commoThe fifth article discussed in this editorial ties together the EMT and TME because of its focus on hypoxia , a hallmSpecifically, authors verify that hypoxia mainly inhibits ERα protein expression. Loss of ERα expression has been associated with endocrine-resistant breast cancer, although this is somewhat controversial. They also observed that a high expression of HIF1α is associated with poor overall survival in ERα-positive patients, which they conclude that this confirms that when solid breast tumors become hypoxic, overall patient survival is reduced. Based on this study, the role of hypoxia in luminal ERα-positive breast cancer emerges as a critical factor interfering with endocrine therapies. Notably, this association is linked to a less favorable clinical prognosis for breast cancer patients .Recent advances in our understanding of the role of various transcription factors in carcinogenesis have provided new opportunities for the development of innovative diagnostic tools and targeted cancer therapies. The identification of TF biomarkers holds promise for early cancer detection and prognosis, while the development of TF-targeted therapies represents a significant leap forward in personalized cancer treatment. As research in this field continues to progress, we can anticipate more effective strategies for treating cancer and improving patient outcomes.


Tissue inflammation is a dynamic process that develops step by step, in response to an injury, to preserve tissue integrity. Resident cells are firstly involved, and once activated, they produce proinflammatory mediators and cytokines that, in turn, activate downstream proinflammatory signals. Blood cells are recruited into the inflammatory environment and activate a phagocytic mechanism to eliminate debris and microorganisms.Chronic tissue inflammation may represent a risk factor for many diseases, such as autoimmune diseases and malignancies. In the past, therapeutic strategies to fight tissue inflammation have mainly been based on the inhibition of proinflammatory mediators and signaling. The resolution of inflammation is an active process that enables inflammation to switch off, protecting tissue from persistent injury and restoring tissue homeostasis. Like inflammation, resolution involves several mediators and signaling pathways that control immune cell functions. A host’s failure to resolve inflammation may be caused by a pathological mechanism driving tissue towards chronic inflammatory conditions. In recent years, the research in this field has been working towards clarifying these pro-resolving pathways and signaling pathways that may be targets for therapeutic intervention ,2.“Signaling Pathways in Inflammation and Its Resolution: New Insights and Therapeutic Challenges” includes five original papers and two reviews in which international experts present new information about novel molecular targets for possible therapeutic strategies to treat inflammatory disease.The Special Issue entitled 2 (PGE2). At the same time, by analyzing the plasma metabolome, they found alterations in lipid metabolism, with a particular increase in fatty acid and cholesterol metabolic pathways, glycerophospholipid metabolism, and sphingolipid metabolism, and a reduction in tricarboxylic acid (TCA) metabolism.In line with recent evidence supporting integrative omics as an important tool for the prognosis and diagnosis of different diseases, Hernandez-Baixauli and coworkers  highlighAnother important issue was addressed by Lee and co-workers . Followi2S is the most recently identified endogenous gas transmitter; it regulates several cell functions, and it is beneficial in several inflammatory conditions and tissue injury. Scheid and coworkers [2S in neuronal injury following the ischemia–reperfusion of the retina in rats. Based on the observation that low doses of H2S may offer neuroprotection, in contrast to high H2S concentrations, which may be toxic [2S inhalation in a rat model of retinal ischemia–reperfusion injury. The results obtained in this study demonstrated that H2S inhalation is beneficial in the ischemia–reperfusion injury of neuronal cells, in a dose- and time-dependent manner, especially at 80 ppm given 1.5 h after reperfusion. The authors showed that a neuroprotective effect is achieved via the inhibition of NF-kB activation, the reduced expression of the pro-inflammatory cytokines IL-1ß and TNF-α, the decreased expression of the pro-apoptotic protein Bax, and the increased expression of the anti-apoptotic Bcl-2.Several eye diseases are associated with ischemia reperfusion injury (IRI). Howorkers  highlighbe toxic ,7, they Antonioli and coworkers  addresseAcute lung injury (ALI) is a clinical disorder with a high mortality rate; together with acute respiratory distress syndrome (ARDS), it is the main cause of respiratory failure associated with high morbidity and mortality rates . Palmito−/− mice. There are data demonstrating that the administration of sCD83 in animal models of inflammation and transplantation leads to the resolution of inflammation and enhances the allograft survival rate. They outlined the function of mCD83 in inflammation as a regulator of DC and Treg and the sCD83 isoform as important in resolving inflammation in autoimmune diseases and in inducing tolerance in transplantation.CD83 is a molecule expressed in a variety of cells. It is present as a membrane-bound isoform (mCD83) highly expressed on activated immune cells, such as dendritic cells (DCs) and B cells, and it is also released as a soluble isoform (sCD83) ,16. HighThe review by Korbecki and coworkers  discusseOverall, these papers provide new insights into the mechanisms of activation of some inflammatory pathways and signaling pathways that could be helpful in the identification of potential therapeutic targets to control inflammation.


A fatal feline panleukopenia virus (FPV) infection occurred in endangered Siberian tigers living in a zoo in the Republic of Korea. Genetic and phylogenetic analyses revealed that the infection was caused by the cross-species transmission of cat-derived FPV prevalent in Korea. The vaccinated tigers were infected with FPV most likely due to insufficient protective immunity or vaccine failure triggered by maternal-derived antibody interference. Therefore, improved biosecurity and vaccination guidelines are required to save the lives of endangered wild carnivores from fatal FPV infection. The data presented in this study will help us expand our knowledge of the molecular epidemiology of FPV in Korea and establish an effective control plan to prevent future FPV infections in captive wild carnivores.Protoparvovirus carnivoran1, is one of the most fatal pathogens of domestic and wild carnivores. The virus endemically infects domestic carnivores worldwide and its cross-species transmission threatens endangered wild carnivores, including Siberian tigers. In this study, a fatal FPV infection in endangered Siberian tigers was investigated to trace the origin of the virus and elucidate the reason behind FPV’s infection of the vaccinated tigers. Our genetic characterization and phylogenetic analysis revealed that the virus detected in the infected tigers, designated as the KTPV-2305 strain, was closely related to FPV strains circulating in Korean cats, suggesting that it might have been transmitted from stray cats wandering around the zoo. Compared with the prototype FPV reference strains, the KTPV-2305 strain carried three distinct amino acid (aa) mutations in the VP2 protein sequence  in this study. These three mutations are commonly found in most global FPV strains, including Korean strains, indicating that these mutations are common evolutionary characteristics of currently circulating global FPVs. The reason why the vaccinated tigers were infected with FPV was most likely the insufficient protective immunity of the affected tigress or vaccine failure triggered by the interference of maternal-derived antibodies in the affected tiger cubs. These findings suggest that improved vaccination guidelines are urgently needed to save the lives of wild carnivores from this fatal virus.Feline panleukopenia virus (FPV), a member of the species  Protoparvovirus carnivoran1, in the genus Protoparvovirus of the family Parvoviridae, along with closely related canine parvovirus 2 (CPV-2) [Feline panleukopenia, caused by feline panleukopenia virus (FPV), is one of the most serious diseases in domestic cats, characterized by anorexia, vomiting, diarrhea, leukopenia, and high mortality in kittens . FPV is  (CPV-2) ,4.As viral DNA synthesis relies on cellular DNA polymerases, FPV replicates in rapidly dividing cells in the intestines, lymphoid tissue, and bone marrow of infected cats, which exhibit clinical presentations of viral infection, such as vomiting, diarrhea, dehydration, lymphopenia, and panleukopenia. The outcome of FPV infection in cats ranges from subclinical to fatal infection with high mortality depending on the cat’s age, immune status, and concurrent infections . DiseaseDue to the ubiquitous distribution of the virus and critical outcomes of the infection, FPV vaccination is highly recommended as a feline core vaccine that all domestic cats should be routinely inoculated with . Both moPanthera tigris altaica), also known as the Amur tiger, is listed as an endangered species by the International Union for the Conservation of Nature. Currently, about 500 Siberian tigers remain in the wild, but many are being bred in zoos around the world [Apart from domestic cats, wild felids are also susceptible to FPV infection, and the clinical outcomes are often fatal, threatening the survival of endangered wild animals such as Siberian tigers ,8. The She world ,10. Althhe world , China [he world , and Korhe world . To protIn May 2023, panleukopenia-suspected clinical signs were observed in captive Siberian tigers at a Korean zoo. The tigers had been vaccinated with commercial FPV MLV vaccines, and one of the infected tigers died after showing severe clinical signs. After the outbreak, an extensive epidemiological investigation was conducted, but the origin and transmission route of the virus have not yet been elucidated. Therefore, the purpose of this study was to identify the origin of this FPV infection through a genetic characterization of the virus and to explore the scientific reason why these tigers were not protected from FPV infection despite being vaccinated with multiple doses of commercial vaccines before the FPV outbreak. The findings of this study expand our knowledge regarding the molecular epidemiology of FPV infection in Korea and establish improved control plans to protect captive wildlife from fatal viral infections, which will contribute to conserving endangered wild animal species.Clinical and epidemiological data of the Siberian tigers were collected to understand the epidemiology of the disease outbreak at Seoul Zoo in Gwacheon city, Gyeonggi-do province, the Republic of Korea. The basic epidemiological information collected included housing practices, vaccination history, clinical manifestation and length of the infection, contacts with other Siberian tigers in the same facility or nearby facilities, and the presence of stray cats roaming around the tigers’ facilities.g for 3 min. Then, total nucleic acids were extracted from 200 µL of each supernatant using a TANBead nucleic acid extraction kit equipped with a fully automated magnetic bead operating platform . The extract was eluted with 100 µL elution buffer according to the manufacturer’s instructions and stored at −80 °C until use. Whole blood samples were collected from each affected tiger via venipuncture of the jugular vein and transferred to a BD Vacutainer® blood collection tube containing anticoagulant . White blood cells (WBCs) were counted using an automatic hematology analyzer  according to the operator’s guide.Fecal samples were collected from five Siberian tigers, including three diseased tigers (two cubs and their mother) and other two Siberian tigers without clinical signs housed in a nearby facility. As the fecal samples to be used in this study were collected by veterinarians at Seoul Zoo without any animal contact, ethical approval from our institute regarding the animals was not required. Prior to our analysis, fecal samples were screened for parvovirus antigens using a commercial immunochromatography assay kit  according to the manufacturer’s protocol. Subsequently, each fecal sample was immersed in 1 mL of phosphate-buffered saline  and then centrifuged at 3000× Molecular analyses were performed on all samples to screen for FPV, feline coronavirus (FCoV), feline calicivirus (FCV), canine distemper virus (CDV), and feline leukemia virus (FeLV) using THUNDERBIRD™ Probe One-step quantitative reverse-transcription polymerase chain reaction (qRT-PCR) kit  according to the manufacturer’s instructions. The primers and probes used in this study are shown in To genetically characterize FPVs detected from Siberian tigers in this study, complete VP2 gene sequences were obtained from two FPV-positive samples with low Ct values. To obtain complete VP2 gene fragment, PCR was performed with a pair of primers  using Takara Ex Taq  as described by An et al. (2011) . A PCR pwww.ncbi.nlm.nih.gov/genebank (accessed on 30 May 2023)). Eighteen Australian FPV sequences  were excluded from this study because their start codons were not definite. The VP2 nucleotide (nt) and aa sequence of the KTPV-2305 strain obtained in this study were aligned with corresponding sequences of the selected 574 global FPV strains, including three FPV reference strains, and four reference CPV-2 strains using MAFFT multiple sequence alignment software (v7.490) [https://www.geneious.com, accessed on 1 June 2023). For the phylogenetic analysis, the IQ-TREE v2.0 software package  was used in this study [The VP2 gene sequences of 595 global FPV strains derived from various hosts were retrieved from the GenBank database ((v7.490) . Based ois study . The phy3/μL) of the affected tiger cubs were 4.89 (Haerang), 3.08 (Sarang), and 0.66 (Parang)  at 7, 12, and 16 weeks of age. The mother tigress had been annually vaccinated with the FPV vaccine since 2011 and the last dose had been administrated in June 2022. Based on the vaccination schedule of the animals in the zoo, the affected siblings were supposed to receive a booster vaccination in May 2023, which was not executed due to the unexpected disease outbreak. However, according to the zoo’s officials, stray cats were often seen roaming around the facilities where the Siberian tigers were housed; thus, there was a potential risk of the virus spreading from stray cats to the tigers through indirect contact or fomites.Prior to the disease outbreak, 13 Siberian tigers were housed alone or in pairs in separate breeding houses with their own playground at Seoul Zoo located in Gwacheon city, Gyeonggi province, the Republic of Korea. As an exception, four tigers including three one-year-old sibling cubs  and a 13-year-old mother tigress (named Penza) were housed in the same breeding house. Starting from 2 May 2023, the four Siberian tigers housed in the same breeding house became sick, showing clinical signs of depression, anorexia, vomiting, diarrhea, nasal secretions, epistaxis, and/or dyspnea . Using a(Parang) , which wOut of the five fecal samples collected from the Siberian tigers, three samples were identified as positive for FPV/CPV via the qPCR assay. All three of the FPV/CPV-positive samples were from clinically ill tigers housed in the same facility, whereas the other two FPV/CPV-negative samples were from clinically healthy tigers housed in a nearby facility . MoreoveTo characterize the FPV detected from diseased tigers, complete VP2 gene sequences were obtained from two positive fecal samples of the affected Siberian tigers. The alignment of the two sequences showed that they were 100% identical to each other, and thereafter, a further analysis was performed with a sequence of the KTPV-2305 strain (GenBank accession number: OR365078). The complete VP2 gene sequence of the KTPV-2305 strain was compared to corresponding sequences of the FPV and CPV-2 reference strains as well as 33 Korean FPV strains available in GenBank . The VP2To further analyze the mutation of the KTPV-2305 strain, the aa sequence of VP2 was compared with that of the reference CPV-2 and FPV, as well as Korean FPV strains derived from cats . The sixn = 5), clade 3 (n = 4), and clade 4 (n = 24). The KTPV-2305 strain detected from the Siberian tigers in this study was grouped into clade 4 and was closely related to previously reported Korean FPV strains detected in domestic or stray cats in 2008, 2017, and 2019 , which were distinct from those of reference FPV strains but were identified in almost all FPV strains throughout the world, including in Korea . AlthougAccording to previous studies, the I101T mutation of FPV strains was first identified in a tiger parvovirus (CHJL-Siberian Tiger-01/2017 strain) isolated from a captive Siberian tiger in China  as well As FPV infection is a fatal disease in domestic cats, several FPV vaccines are commercially available and widely used as core vaccines according to the guidelines for feline vaccination recommended by the World Small Animal Veterinary Association (WSAVA) ,42. HoweIn this study, a fatal FPV infection in endangered Siberian tigers was investigated to track the origin of the virus and elucidate the reason behind the FPV infection of the tigers despite them being vaccinated against the virus. Genetic characterization and a phylogenetic analysis revealed that the virus detected in the diseased tigers was closely related to FPV strains prevalent in Korean cats, suggesting that the virus was transmitted from the stray cats roaming around the zoo to the tigers. The tigers were infected with FPV despite receiving multiple doses of a commercial vaccine most likely due to insufficient protective immunity or vaccine failure triggered by MDA interference, indicating that more effective vaccination guidelines are needed to save wild carnivores from fatal viral pathogens. The findings of this study will help expand our knowledge of the molecular epidemiology of FPV in Korea and establish control measures to prevent future FPV infections in captive wild carnivores.