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Sticholysin II (StII), a pore-forming toxin from the marine anemone Stichodactyla helianthus, enhances an antigen-specific cytotoxic T lymphocyte (CTL) response when co-encapsulated in liposomes with a model antigen. This capacity does not depend exclusively on its pore-forming activity and is partially supported by its ability to activate Toll-like receptor 4 (TLR4) in dendritic cells, presumably by interacting with this receptor or by triggering signaling cascades upon binding to lipid membrane. In order to investigate whether the lipid binding capacity of StII is required for immunomodulation, we designed a mutant in which the aromatic amino acids from the interfacial binding site Trp110, Tyr111 and Trp114 were substituted by Ala. In the present work, we demonstrated that StII3A keeps the secondary structure composition and global folding of StII, while it loses its lipid binding and permeabilization abilities. Despite this, StII3A upregulates dendritic cells maturation markers, enhances an antigen-specific effector CD8+ T cells response and confers antitumor protection in a preventive scenario in C57BL/6 mice. Our results indicate that a mechanism independent of its lipid binding ability is involved in the immunomodulatory capacity of StII, pointing to StII3A as a promising candidate to improve the reliability of the Sts-based vaccine platform., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Growing evidence suggests that opioid analgesics modulate angiogenesis during pathophysiological processes. Vascular endothelial growth factor-A (VEGF-A) was recently proposed to be involved in pain development. To date, no anti-angiogenic drug is used for pain management. When administered in a bioavailable formulation, (i.e., ultramicronized) N-palmitoylethanolamine (PEA) delays the onset of morphine tolerance, improves morphine analgesic activity and reduces angiogenesis in in vivo models. This study aimed at investigating whether VEGF-A is involved in PEA-induced delay of morphine tolerance. The anti-VEGF-A monoclonal antibody bevacizumab was used as a reference drug. Preemptive and concomitant treatment with ultramicronized PEA delayed morphine tolerance and potentiated the analgesic effect of morphine, while counteracting morphine-induced increase of VEGF-A in the nervous system. Similar results were obtained when bevacizumab was administered together with morphine. Of note, bevacizumab showed an analgesic effect per se. In equianalgesic treatment regimens (obtained through increasing morphine doses and associating PEA), PEA resulted in lower expression of VEGF-A in dorsal root ganglia (DRG) and spinal cord compared to morphine alone. Similar results were observed for plasma levels of the soluble VEGF receptor 1 (sFLT-1). Moreover, in morphine-treated animals, two pain-related genes (i.e., Serpina3n and Eaat2) showed a more than 3-fold increase in their expression at spinal cord and DRG level, with the increase being significantly counteracted by PEA treatment. This study supports the hypothesis that the effects of PEA on morphine analgesia and tolerance may be mediated by the down-modulation of VEGF-A and sFLT-1 in the nervous system and plasma, respectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Carla Ghelardini and Lorenzo Di Cesare Mannelli received a grant from Epitech (Padua, Italy)., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Oxidized albumin is considered a short-term biomarker of oxidative stress and its measurement in blood contributes to evaluate the impact of diseases, drugs, dialytic treatments, physical activity, environmental contaminants etc. on the red-ox balance of humans as well as of other mammalians. Nevertheless, the most common methods for quantifying the oxidized and reduced albumins are costly and time-consuming. Furthermore, there is a dearth of information regarding the proper ways to store human serum or plasma samples in order to prevent inaccurate quantification of these various albumin forms. This paper explores these aspects and proposes a few spectrophotometric assay procedures which make the quantitation of oxidized and reduced albumin very fast, precise and un-expensive in various mammals., (Copyright © 2024. Published by Elsevier Inc.)

An Enzyme Linked ImmunoMagnetic Electrochemical assay (ELIME) was developed for the detection of the hepatitis A virus (HAV). This system is based on the use of new polydopamine-modified magnetic nanobeads as solid support for the immunochemical chain, and an array of 8 screen-printed electrodes as a sensing platform. Enzymatic-by-product is quickly measured by differential pulse voltammetry. For this purpose, all analytical parameters were optimized; in particular, different blocking reagents were evaluated in order to minimize the nonspecific interaction of bioreagents. Using the ELIME assays, a quantitative determination of HAV can be achieved with a detection limit of 1·10 -11 IU mL -1 and a working range between 10 -10 - 5 × 10 -7 IU mL -1 . The cross-reactivity of the commercial monoclonal antibodies against HAV used in ELIME assays was tested for Coxsackie B4, resulting very low. The sensitivity was also investigated and compared with spectrophotometric sandwich ELISA. The average relative standard deviation (RSD) of the ELIME method was less than 5% for the assays performed on the same day, and 7% for the measurements made on different days. The proposed system was applied to the cell culture of HAV, which title was quantified by Real-Time Quantitative Reverse Transcription PCR (RT¬qPCR). To compare the results, a correlation between the units used in ELIME (IU mL -1 ) and those used in RT¬qPCR (genome mL -1 ) was established using a HAV-positive sample, resulting in 1 IU mL -1 -10 -4 gen mL -1 (R 2  = 0.978). The ELIME tool exhibits good stability and high biological selectivity for HAV antigen detection and was successfully applied for the determination of HAV in tap water., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Vegetable wastes represent an inexpensive and sustainable source of valuable bioproducts for several applications. Natural micro-porous and fibrous materials can be obtained from a very cheap and abundant cellulosic bio-waste. Here we demonstrated that vegetable waste derivatives can be suitable as scaffolds for biosensors and 3D cell growth. Many studies have been addressed to fabricate biocompatible 3D scaffolds for mammalian stem cells cultures and develop novel systems able to reproduce the complexity of the in vivo microenvironment. Many of these products are proprietary, expensive or require chemical synthesis. The recycling and revaluation of vegetable derived tissues to fabricate scaffolds for analytical biosensors 3D stem cell cultures platforms may represent a very low-cost approach for toxicological and environmental analyses. In this approach, potential applications of vegetable-derived tissue for biosensing and 3D stem cell cultures were investigated. Micro-structured scaffolds from stalk of broccoli, named BrcS, were either functionalized for production of enzymatic 3D-biosensors or preconditioned to be used them as 3D-scaffolds for human mesenchymal stem cells cultures. The conditions to fabricate 3D-biosensors and scaffolds for cell growth were here optimized studying all analytical parameters and demonstrating the feasibility to combine these two properties for an innovative solution to ennoble vegetable wastes., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Hydrogel-based cleaning of paper artworks is an increasingly widespread process in the cultural heritage field. However, the search for tuned (compatible, highly retentive and not perishable) hydrogels is a challenging open question. In this paper, a complete characterization of chemical hydrogels based on polyvinyl alcohol (PVA) crosslinked with telechelic PVA and their remarkable performances as gels for cleaning paper artworks are reported. The rheological properties, porosity, water content of these gels were determined and analyzed as a function of the components concentration during synthesis. Due mechanical and retentive properties, the reported gels are optimum candidates for paper cleaning applications. The efficacy of these PVA-based gels has been demonstrated applying them on the surface of the sheets of several paper artworks, and characterizing the samples before and after the cleaning process by means of a multidisciplinary approach involving spectroscopic and chromatographic tests., Competing Interests: Declaration of Competing Interest All authors declare no conflicts of interest and no competing financial interest, (Copyright © 2020 Elsevier B.V. All rights reserved.)

Cortisol is a steroidal hormone and an important stress marker. Free serum cortisol concentration has been identified to correlate well with free salivary cortisol. In this present work an electrochemical immunosensor was developed to determine cortisol concentration within the physiological concentration range found in human saliva. The immunosensor is based on a direct competitive enzyme linked immunoassay using a home-made cortisol-alkaline phosphatase (AP) conjugate synthesized in our laboratory with disposable graphite screen-printed electrodes (SPEs). 1-nalphtyl phosphate (1-NP) was used as an enzymatic substrate and a square wave voltammetry (SWV) for electrochemical detection. To study method suitability for use with saliva samples, calibration curves were performed both in buffer and saliva. In buffer standard samples showed a limit of detection (LOD) of 0.6 ng/ml and working range (WR) of 0.2-44.6 ng/ml with good reproducibility (RSD 10%). Saliva matrix effect was removed effectively with Salivette Cortisol collection device (polyethylene) and a calibration curve showed similar characteristics as in buffer with LOD 1.7 ng/ml and WR 0.5-55.1 ng/ml (RSD 8%) demonstrating the possibility to determine human salivary cortisol within the desired human physiological range. Spiked saliva samples were analyzed with the developed immunosensor presenting excellent 92-114% recovery. Comparison to liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method showed strong 0.90 correlation between methods indicating good accuracy of the developed immunosensor., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background and Aims: Anthracyclines are effective anticancer drugs that have improved prognosis of hundred thousand cancer patients worldwide and are currently the most common chemotherapeutic agents used for the treatment of blood, breast, ovarian and lung cancers. However, their use is limited because of a cumulative dose-dependent and irreversible cardiotoxicity that can cause progressive cardiomyopathy and congestive heart failure. Aim of the present study was to determine the cardioprotective activity of a dietary source of cyanidin 3-glucoside (C3G), such as purple corn, against doxorubicin (DOX)-induced cardiotoxicity in mice., Methods and Results: In vitro studies on murine HL-1 cardiomyocytes showed that pretreatment with both pure C3G and purple corn extract improved survival upon DOX treatment. However, C3G and purple corn extract did not affect the cytotoxic effect of DOX on human cancer cell lines. We then validated in vivo the protective role of a C3G-enriched diet against DOX-induced cardiotoxicity by comparing the effect of dietary consumption of corn isogenic lines with high levels of anthocyanins (purple corn - Red diet - RD) or without anthocyanins (yellow corn - Yellow diet - YD) incorporated in standard rodent diets. Results showed that mice fed RD survived longer than mice fed YD upon injection of a toxic amount of DOX. In addition, ultrastructural analysis of hearts from mice fed RD showed reduced histopathological alterations., Conclusion: Dietary intake of C3G from purple corn protects mice against DOX-induced cardiotoxicity., (Copyright © 2017 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

Cerebellar granule neurons develop postnatally from cerebellar granule precursors (GCPs), which are located in the external granule layer (EGL) where they massively proliferate. Thereafter, GCPs become postmitotic, migrate inward to form the internal granule layer (IGL), further differentiate and form synapses with Purkinje cell dendrites. We previously showed that the Btg family gene, Tis21/Btg2, is required for normal GCP migration. Here we investigated the role in cerebellar development of the related gene, Btg1, which regulates stem cell quiescence in adult neurogenic niches, and is expressed in the cerebellum. Knockout of Btg1 in mice caused a major increase of the proliferation of the GCPs in the EGL, whose thickness increased, remaining hyperplastic even after postnatal day 14, when the EGL is normally reduced to a few GCP layers. This was accompanied by a slight decrease of differentiation and migration of the GCPs and increase of apoptosis. The GCPs of double Btg1/Tis21-null mice presented combined major defects of proliferation and migration outside the EGL, indicating that each gene plays unique and crucial roles in cerebellar development. Remarkably, these developmental defects lead to a permanent increase of the adult cerebellar volume in Btg1-null and double mutant mice, and to impairment in all mutants, including Tis21-null, of the cerebellum-dependent motor coordination. Gain- and loss-of-function strategies in a GCP cell line revealed that Btg1 regulates the proliferation of GCPs selectively through cyclin D1. Thus, Btg1 plays a critical role for cerebellar maturation and function., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Two new protein conjugates were prepared and studied to develop and compare two (direct and indirect) competitive enzyme-linked immunosorbent assay (ELISA) formats for the determination of cortisol in human saliva. Toward this goal, ovalbumin was conjugated to cortisol and used for developing an indirect competitive ELISA, while alkaline phosphatase was coupled with the same analyte for a direct competitive assay. The yield of the conjugation reactions was evaluated. The results obtained show that the indirect and direct ELISA formats developed for cortisol had working ranges of 0.5-70 and 2-330 ng/ml and detection limits of 0.5 and 1.2 ng/ml, respectively. Artificial and real saliva samples were spiked with cortisol to study the matrix effect of saliva. The suitability of the assays for quantification of cortisol in saliva was also studied., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The present review deals with novel developments in immunosensors destined for final application in food analysis. In this perspective particular emphasis will be given to the most important approaches which recently have been used for immunosensor construction and assembling. For this reason, electrochemical, surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) techniques will be explored in detail and recent and practical examples on food matrices will be reviewed. Objective of this survey is to give a general overview of the possible application of immunosensors to the food analysis field.

Objective: Corticotropin-releasing factor (CRF) is a placental neuropeptide that plays a role in the control of uteroplacental blood flow regulation. Because CRF has a relaxant effect on uterine vasculature in pregnant rats, we aimed to evaluate mid-gestation plasma CRF levels in women with impaired uterine artery blood flow., Methods: Maternal plasma CRF was assayed by specific radioimmunoassay, and uterine artery resistance index (RI) was assessed by Doppler evaluation at 22-24 weeks' gestation in 55 healthy pregnant women, of whom 24 showed a unilateral or bilateral uterine artery notch, reflecting resistance. Statistical analysis was performed by the Kruskal-Wallis test followed by the post hoc Dunn's test and the Spearman rank test., Results: The mean uterine artery RI was significantly (P <.001) higher in women with a notch than in healthy controls. Mean +/- standard error of the mean maternal plasma CRF levels were significantly lower in women with a unilateral (168.45 +/- 27.5 pg/mL; P <.01) or bilateral (186.07 +/- 34.5 pg/mL; P <.001) uterine artery notch than in healthy control pregnant women (375.06 +/- 21.77 pg/mL). Although no difference was found in CRF levels between patients with a unilateral or bilateral uterine artery notch, a significant inverse correlation was found between the mean RI and maternal plasma CRF levels (Spearman r = -0.6540; 95% confidence interval, -0.7865, -0.4640; P <.001)., Conclusion: Reduced levels of circulating CRF were associated with increased uterine artery resistance, which supports the hypothesis that CRF may regulate uterine artery tone at mid gestation.

With the explosion of the number of youth participating in sports activities has come an epidemic of injuries. Myriad factors make their prevention a great challenge for society. This article presents an overview of statistics, factors contributing to sports injuries, a review of significant prevention efforts, barriers to prevention, and recommendations for the future.

The injuries that are prevalent in and unique to dancers have their origins inextricably linked to faulty technique or poor biomechanics, combined with other risk factors. It is this combination of factors that must be addressed when considering diagnosis, treatment, and rehabilitation. For the dancer to return to full activity with minimal risk of recurrent injury, neuromuscular re-education is mandatory. This process best entails using a team approach, accessing the resources of healthcare professionals and those who train the dancer on a daily basis. In some cases, parents also may need to be involved. Communication, interaction, and mutual understanding among these groups will assist the dancer in regaining and maintaining health.

The diagnosis of back pain in the young athlete should be specific and not attributed to nonspecific, mechanical causes. Risk factor identification and intervention are required. Treatment is then initiated in a specific pattern, addressing flexibility and muscular imbalances. Bracing is often used to allow healing of growth tissue. The lumbosacral orthosis may be molded in a lordotic posture to unload the disc or antilordotic posture to relieve the posterior column; however, customizing the lordosis to the individual biomechanics may be required. Spinal stabilization is initiated with therapy for strengthening isolated weaknesses and progressing to coactivation and proprioceptive techniques, such as the balance ball. Returning to competition is preceded with sport-specific training.

The youth soccer movement in the United States is developing at a tremendous rate. This article explores the epidemiology of youth soccer including trends in participation, injury data, and injury categories that are of specific concern to the young athlete. Finally, recommendations are made regarding possible future areas of study and research on youth soccer.

Stress fractures are a source of significant sports disability in the growing athlete. Early diagnosis and treatment are paramount in the management of these injuries to minimize the morbidity associated with them. In addition, the identification of potential host and/or environmental risk factors, and education of athletes, parents, coaches and physicians about these risk factors are key in the prevention of stress fractures and other overuse injuries in the pediatric athlete.

A simple high-performance liquid chromatographic (HPLC) method for the determination of flufenamic acid in rat plasma is described. After liquid-liquid extraction, the drug is separated by HPLC on a 5-microns octadecylsilica column (Nucleosil C18) with ultraviolet detection at 280 nm. Linear calibration graphs for flufenamic acid were constructed from 0.5 to 15 micrograms/ml. The method has been applied to a pharmacokinetic study in animals.

Specific elbow and wrist injuries are predictable in the skeletally immature athlete based on the biomechanics of the sport and the age of the patient. The physician must be aware of the potential for overuse injuries. Modification in training regimens is essential for recovery. A greater emphasis must be placed on the prevention of these injuries. As a general rule, the young athlete should not progress more than 10% per week in the amount and frequency of training. Correction of muscle-tendon imbalances is accomplished by maintaining strength and flexibility of susceptible tissues. In throwers, a triceps-strengthening program of progressive resisted extension exercises and a forearm flexor/extensor-strengthening program using the French curl technique are helpful. Careful attention to throwing technique and proper coaching are essential. The goal for the young athlete is early recognition of the injury and thereby prevention of a long-term disability.

Overuse injuries in the young athlete now are seen more frequently. Using an adult model of these injuries with respect to diagnosis and treatment and applying it to the young athlete does not address many of the unique characteristics that contribute to these injuries. An overview of causative factors that contribute to overuse injuries in young athletes is presented. Examples of specific injuries seen only in this age group and the treatment of these injuries are presented.

Back pain in children and young athletes is very different from back pain in adults. Macrotrauma must be carefully evaluated and managed, even in the absence of definitive radiographic findings. Microtrauma must be suspected in at-risk athletes. These athletes require persistent diagnostic evaluation and may require SPECT bone scan to uncover a posterior element stress reaction. Atraumatic back pain requires the elimination of neoplastic, infectious, rheumatologic, or congenital causes.

Certain topics and controversies involving young athletes are addressed, including unclear criteria for volume and intensity of training and acute and overuse injuries of the knee. Osteochondritis dissecans, tibial spine avulsions, peri-pelvic apophyseal avulsions, and back injuries are discussed.

As a fraction of ingested ethanol is metabolized by gastric mucosa, different amounts of alcohol should reach the liver when the same dose is administered by oral or intravenous route. Therefore, we investigated the time-course of hepatic reduced glutathione (GSH) concentrations after intra-peritoneal or intra-gastric load of the same amount of ethanol in the rat. The test was also performed in fasted and Cimetidine-treated rats. The oral ethanol administration was followed by a less pronounced decrease and by a quicker recovery of hepatic content of GSH than after intraperitoneal route. In the fasted rat, basal hepatic GSH significantly decreased; after alcohol administration the decrease of hepatic GSH was more severe and prolonged than in the fed rat. Cimetidine was shown to be a potent inhibitor of gastric ADH. Pre-treatment with Cimetidine did not change the basal levels of hepatic GSH, but after oral ethanol load, the decrease of the hepatic GSH content was significantly (p < 0.005) more pronounced than in controls. This study demonstrates the beneficial effects of gastric ethanol metabolism on the liver. The reduced gastric ethanol metabolism, induced by fasting or by Cimetidine resulted in a decreased content and delayed recovery of liver GSH content.

A rapid and precise high-performance liquid chromatographic method for the determination of piroxicam in a variety of biological samples has been developed. A reversed-phase column, isocratic elution and ultraviolet detection were employed. Calibration curves were reproducible and highly linear, with correlation coefficients typically averaging over 0.992. The detection limit of the assay was 100 ng/ml for all biological samples examined (at a signal-to-noise ratio of 3:1). Validation of the method demonstrated a good sensitivity, accuracy and precision. The method has been adopted for a pharmacokinetic study in rats.

Overuse injuries to tendons and apophyses are being seen with increasing frequency in children and adolescents as they increase their participation in organized sports, fitness, and dance activities. A retrospective review of 724 overuse tendon and apophyseal injuries seen in the Sports Medicine Clinic at the Boston Children's Hospital suggested that growth and maturation constitute additional risk factors for injury occurrence. Diagnosis and treatment of these injuries is discussed in the context of sports participation.

The prospect of the application of scientific techniques of physiology and movement science to dance promises to enhance not only the performance but also the safety of ballet dancing. The scientific analysis of nutrition, strength, and cardiovascular and movement demands of elite athletes in a great variety of competitive sports has increased our understanding of the stresses placed upon the body in these sports and in so doing has improved training techniques, enhanced performance, and helped to decrease the incidence of injuries. The application of these scientific analysis techniques to dance may reap similar benefits. Although the classical ballet teacher may, at first glance, be horrified by a computer analogue analysis of a grand jete, the information obtained from such studies may help him or her to decide when a young dancer is ready for such a technique or demonstrate a technical deficiency in the performance of such a technique. Similarly, the often debated question of when a young dancer should attempt pointe may become less subject to a teacher's intuition or parental pressure if research on the strength of muscles, bones, and ligaments teaches us the specific levels needed before this technique should be attempted.

The traction apophysitises are one of the frequently encountered overuse injuries to children. Although known for many years and classed as osteochondroses, the traumatic origin of these disorders--from repetitive microtrauma--is now receiving wider acceptance. Discussion of the assessment, treatment, and prevention of these disorders is included.

Overuse injuries are now well known to sports enthusiasts at any age or level of competition. The seeming explosion of overuse stress fractures of lower extremity bones in high-profile professional basketball players has brought about widespread media attention and a better understanding of the phenomenon of "overuse syndrome" by the public. However, the spectrum of overuse injuries in the child or adolescent athlete has only recently been recognized. These injuries can range from the permanent disability of osteochondritis dissecans of the elbow to the completely nonspecific "growing pains" of active youngsters.

The primary purpose of a preparticipation physical examination is to identify risk factors that may predispose an athlete to physical and/or psychological injury. Inclusion of a physiological assessment complements the more traditional preparticipation health examination by contributing valuable information toward the specific physical strengths and weaknesses of the young athlete. This information when combined with the orthopedic and medical components of a preparticipation examination can dramatically enhance the safety of sports participation.

Childhood participation in sports involves a complex interaction of many physical, psychological, and social factors. Some children and teenagers cope well with the stress of performance and competition and their participation leads to enhanced self-esteem and personal growth; others cope poorly, however, and are at a higher risk for injury. Based on clinical observations of patients seen through the sports and orthopedic services at a major children's hospital, this article has identified two groups of children and adolescents at risk for psychological complications following injury. Psychotherapeutic intervention is seen as a fundamental component in the overall treatment planning for patients manifesting a difficult recovery course, prolonged interference in athletic injury, and a tendency toward reinjury.

The complaint of low back pain in the adolescent must never be taken lightly. A high index of suspicion should be particularly entertained in a child participating in gymnastic training or competition. As noted in this article, steps can now be taken, particularly if a specific diagnosis is made early, to institute specific treatment with a high likelihood of success. Young gymnasts complaining of back pain must never be passed off as having sustained a back strain or "muscle spasms" and treated symptomatically. Persistent back pain beyond two weeks warrants, in our opinion, a complete evaluation, careful history and physical examination, a four-view radiographic assessment of the spine, and, if necessary, bone scans or other more advanced techniques to make a specific diagnosis of the cause of the pain.

Back injuries may occur at any age or level of skill and are slightly more prevalent in male dancers. The risk factors that may be involved in a given injury include training errors, musculotendinous imbalance, anatomic malalignment of the lower extremity, shoe wear, and floor surface. Assessment of these risk factors provides guides for management and prevention.