Determination of phenolic compounds in human saliva after oral administration of red wine by high performance liquid chromatography.

Red wine is a relevant source of bioactive compounds, which contribute to its antioxidant activity and other beneficial advantages for human health. However, the bioavailability of phenols in humans is not well understood, and the inter-individual variability in the production of phenolic compounds has not been comprehensively assessed to date. The present work describes a new method for the extraction and analysis of phenolic compounds including gallic acid (Gal), vanillic acid (Van), caffeic acid (Caf), syringic acid (Sir); (-)-epicatechin (Epi); p-coumaric acid (Cum) and resveratrol (Rsv) in human saliva samples. The target analytes were extracted using Fabric Phase Sorptive Extraction (FPSE), and subsequently analysed by high-performance liquid chromatography (HPLC) coupled with photodiode array detector (PDA). Chromatographic separation was achieved using a Symmetry C18 RP column in gradient elution mode, with methanol and phosphate buffer as the mobile phases. The linearity (intercept, slope, and determination coefficient) was evaluated in the range from 1 to 50 µg/mL. The limit of quantification (LOQ) was 1 µg/mL (LLOQ ≥0.8 µg/mL), whereas limit of detection was 0.25 µg/mL. The intra and inter-day RSD% and BIAS% values were less than± 15%. The analytical performances were further tested on human saliva collected from healthy volunteers after administering red wine. To the best of our knowledge, this is the first FPSE procedure for the analysis of phenols in saliva, using a non-invasive and easy to perform sample collection protocol. The proposed fast and inexpensive approach can be deployed as a reliable tool to study other biological matrices to proliferate understanding of these compounds distribution in human body.

Fast liquid chromatography-tandem mass spectrometry method for the simultaneous determination of phytocannabinoids in oily based preparations.

The reported method aims to be a powerful aid for the simultaneous determination of tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), cannabigerol (CBG), tetrahydrocannabinolic acid (THCA), cannabidiolic acid (CBDA), and tetrahydrocannabivarin (THCV) in oily based preparations. The chromatographic separation was carried out using an Hypersil Gold PFP (50 × 2.1 mm, 1.9 µm) column, using H2O + 2 mM ammonium formate + 0.2 % formic acid (M1) and Methanol + 2 mM ammonium formate + 0.2 % formic acid (M2) as mobile phases. The flow rate was set 0.4 mL/min. Specifically, this method was validated in terms of linearity, limit of detections and quantifications (LODs and LOQs), accuracy (precision and trueness, both intra and interday), selectivity, and matrix effects. This procedure allowed quantifying seven phytocannabinoids in less than 10 min. The validated method shows a good linearity within the range 0.25-1000 ng/mL, while precision and trueness (intra- and inter-day) were below <13.25 % and 7.59 %, respectively. Regarding the matrix effect, the method satisfies all the requirements, except for the THC and THCV, where it reaches about 120 %. This element does not affect the method performances as it has been observed that this value is constant and reproducible and therefore does not involve errors in the quantitative analysis. The method was tested and applied on more 70 different oily based preparations. Furthermore, starting from four different cannabis cultivar (FM2, Bedrolite, Bedrocan, and Bediol), it allowed to evaluate the reproducibility of the magistrali preparations. The real samples, in fact, derive from different local pharmacies, and were analyzed by the accredited UNI CEI EN ISO/IEC 17025:2018, Pharmatoxicology Laboratory (ACCREDIA, lab n. 2274 ASLPE, accreditation number 1822 L), accordingly to the current regulations.

Fast off-line FPSE-HPLC-PDA determination of six NSAIDs in saliva samples.

A fast off-line FPSE-HPLC-PDA method has been reported that allows simultaneous clean up and determination of six non-steroidal anti-inflammatory drugs (NSAIDs) in saliva samples from healthy volunteers. Particularly, furprofen, indoprofen, ketoprofen, fenbufen, flurbiprofen, and ibuprofen were chromatographically resolved. Benzyl paraben was chosen as the internal standard (BzPB, IS). These target compounds were successfully extracted from human saliva using fabric phase sorptive extraction (FPSE) and then analysed in the liquid chromatographic system by means of a short analytical column (Symmetry C18, 75 × 4.6 mm, 3.5 µm) using acetonitrile (AcN) and phosphate buffer (PBS, 30 mM; pH = 2.5) as the mobile phases. The method, validated through the calculation of all analytical parameters in accordance of International Guidelines, was applied to real saliva sample analysis collected from informed volunteers. The proposed approach that included the use of sol-gel polytetrahydrofuran (sol-gel PTHF) sorbent immobilized on cellulose support and C18 stationary phase used in HPLC, showed high potential as a fast tool for future clinical and forensic applications. The herein reported results encourage potential future application of FPSE in the forensic field. Furthermore, the FPSE membrane was tested in dried saliva spot mode (DSS) in order to check its potential use as a sampling device, also for forensic applications.

Stimulated mast cells release inflammatory cytokines: potential suppression and therapeutical aspects.

Mast cells (MCs) are derived from bone marrow precursors and are immune cells involved in acute and chronic inflammation. MCs are ubiquitous and play a crucial role in innate and acquired immunity. They are activated through cross-linking of their surface high affinity receptors (FcεRI), leading to immediate secretion of stored inflammatory mediators, and late production and release of pro-inflammatory cytokines/chemokines without degranulation. Therefore, MCs are important in inflammatory responses. Members of the interleukin (IL)-1 cytokine family, such as IL-1 and IL-33, and various antigens markedly increase IL-1 and tumor necrosis factor (TNF) expression and secretion from MCs. One of the latest cytokines is IL-33, an IL-1 family member acting via its ST2/IL-1R4, which has been shown to regulate MCs. IL-1 and IL-33 are cytokines found to be implicated in many inflammatory disorders including rheumatoid arthritis, atherosclerosis and psoriasis. In general, IL-1 family member cytokines play a pro-inflammatory role and increase the pathological state. IL-37 is a member of the IL-1 family with anti-inflammatory activity through inhibition of pro-inflammatory cytokines. IL-37 particularly suppresses IL-1-mediated innate inflammatory response, but also acts on the acquired immune response. IL-37 is activated by pro-inflammatory agents and cytokines, playing a protective role against inflammation. This cytokine is a natural regulator of immunity and is a therapeutic promise against inflammatory diseases. Since IL-1 is produced by and activates MCs to release IL-33 and TNF, here we hypothesize that MCs can be inhibited by IL-37 and therefore reduce their pro-inflammatory activity. However, the maturation, transport and secretion of IL-37 remain to be clarified.

IL-33 mediates allergy through mast cell activation: Potential inhibitory effect of certain cytokines.

Mast cells (MCs) are hematopoietic immune cells commonly found in adjacent to blood vessels in the lamina propria of airway mucosa. They are important in allergic reactions since the cross-linking of their surface high affinity receptor FceRI induces activation of these cells, and provokes the synthesis, degranulation and release of inflammatory mediators including arachidonic acid-derived eicosanoids (de novo synthesized), stored enzyme mediators, and inflammatory TH1 and TH2 cytokines, and chemokines. Interleukin (IL)-33 participates in innate and adaptive immunity and inflammation and, acting on CD34+ cells, causes MC differentiation and maturation. IL-33 is generated by activated immune cells, and activates MCs which degranulate and release pro-inflammatory mediators. IL-33 is very important in mediating allergic inflammation and can be induced by IL-1 beta. It is also called "alarmin" and is an inflammatory cytokine IL-1 family member, expressed from mocytes and MCs, which binds its receptor ST2, provoking its release after cell damage. MC-derived allergic compounds in response to IL-33 is critical to innate type 2 immunity. IL-37 is expressed by immune and non-immune cells after pro-inflammatory stimulus. IL-37, an anti-inflammatory cytokine, binds IL-18Ra and suppresses pro-inflammatory IL-1 beta released by activated immune cells such as macrophages. Here, we hypothesize that pro-inflammatory IL-1 family member cytokines released by activated MCs, mediating inflammatory allergic phenomenon, can be suppressed by IL-37.

Impact of mold on mast cell-cytokine immune response.

Molds include all species of microscopic fungi, the spores of which are small molecules, ubiquitous, mostly found in soil with higher rainfall and high humidity, in the atmosphere of urban and rural settings and in decaying vegetation. They originate from pathogenic fungi and have a crucial role in inflammatory response, causing a broad range of diseases. Immune suppressed subjects may develop mycoses caused by opportunistic common pathogenic fungi. Mast cells (MCs) are immune cells involved in the pathophysiology of infected skin, lung, and organs, where there is an increase of angiogenesis. Airways fungi infections can induce allergic lung disease mediated by MCs and other immune cells. In addition, fungal infection may cause and/or aggravate asthma inflammation. Spores are able to navigate in the airways of the lung and can be recognized trough toll-like receptor (TLR) signaling by the innate immune cells including MCs. Activated MCs release preformed mediators including histamine, proteases (tryptase, chimase), pro-inflammatory cytokines/chemokines and they also generate arachidonic acid products. MCs activated by fungi provoke an increases of PGD2 levels and lead to hypersensitivity diseases which present signs such as irritation of the respiratory tract and eyes, recurrent sinusitis, bronchitis, cough and neurological manifestations including fatigue, nausea, headaches and brain fog. Therefore, fungi activate the innate immune response through the TLRs, leading to the release of myeloid differentiation factor 88 (MyD88) which, with a series of cascade reactions, induces the stimulation of AP-1 and NF-kB with subsequent activation of inflammatory IL-1 family members. Here, we report that fungi can activate MCs to secrete pro-inflammatory cytokines which may be inhibited by IL-37, a new anti-inflammatory IL-1 family member.

New concepts in neuroinflammation: mast cells pro-inflammatory and anti-inflammatory cytokine mediators.

The activation of brain nociceptors and neurons may lead to neurogenic inflammation, an event that involves immune cells including mast cells (MCs). Microglia are similar to macrophages and secrete pro-inflammatory IL-1 family members and TNF. TNF is rapidly released (first 10 minutes from MC granules) and is subsequently secreted along with other pro-inflammatory cytokines with a new synthesis after several hours. MC-derived TNF is a very powerful pro-inflammatory cytokine which mediates sensitization of the meningeal nociceptors. Here, we report the involvement of MCs in neuroinflammation, the role of inflammatory cytokine IL-1 family members, and of TNF, as well as the potential inhibition of IL-37.

Low-grade chronic inflammation mediated by mast cells in fibromyalgia: role of IL-37.

It has been observed that acute stress causes the activation of TH1 cells, while TH2 cells regulate and act on chronic inflammation. Fibromyalgia (FM) is a chronic, idiopathic disorder which affects about twelve million people in the United States. FM is characterized by chronic widespread pain, fatigue, aching, joint stiffness, depression, cognitive dysfunction and non-restorative sleep. The mechanism of induction of muscle pain and inflammation is not yet clear. In FM there is an increase in reactivity of central neurons with increased sensitivity localized mainly in the CNS. Mast cells are involved in FM by releasing proinflammatory cytokines, chemokines, chemical mediators, and PGD2. TNF is a cytokine generated by MCs and its level is higher in FM. The inhibition of pro-inflammatory IL-1 family members and TNF by IL-37 in FM could have a therapeutic effect. Here, we report for the first time the relationship between MCs, inflammatory cytokines and the new anti-inflammatory cytokine IL-37 in FM.

Legal aspects in implantable defibrillator extraction.

At the Institute of Legal Medicine in Chieti, a case of iatrogenic superior vena cava perforation was observed during laser extraction of an infected biventricular implantable cardiac defibrillator. The presentation of this particular case represented a starting point for studying the occurrence of similar complications in literature, since their knowledge and understanding should induce resolution of any organisation problems, aid in increasing physicians' training and impose the availability of cardiac surgeons during such operations.

First report of an acute purulent maxillary sinusitis caused by Pseudomonas aeruginosa secondary to dental implant placement in an immunocompetent patient.

STUDY DESIGN: In this case report, we present maxillary Pseudomonas aeruginosa sinusitis in an immunocompetent patient who underwent an autologous bone transplant for the insertion of dental implants. RESULTS: The infection was eradicated after removal of the dental implants and long-term antibiotic therapy. CONCLUSION: Despite the infection resolution, severe complications were observed with important legal consequences.

Diagnostic failure of ciprofloxacin-induced spontaneous bilateral Achilles tendon rupture: case-report and medical-legal considerations.

Rare side-effects of fluoroquinolone therapy are tendinitis and tendon rupture. Many reports have demonstrated that the concomitant use of corticosteroids, in patients aged 60 years or older, increase the risk substantially. We present a case of spontaneous bilateral Achilles tendon rupture induced by ciprofloxacin and methylprednisolone. A 61-year-old woman was diagnosed with Bronchiolitis Obliterans with Organizing Pneumonia (BOOP) and was started on oral ciprofloxacin 500 mg twice daily for 3 weeks and on oral methylprednisolone 16 mg twice daily for 2 weeks. The diagnosis was made after doctors, rather than stop drug therapy and advise complete rest, had mistakenly prescribed for the woman to undergo physiotherapy and local NSAIDs, thus favoring the onset of tendon ruptures and resulting in surgical and legal implications. Inspired by this case, we also submit a brief review on professional liability in Orthopaedics.

A case of accidental aspiration of a dental cutter into the bronchopulmonary tree: clinical implications and legal considerations.

Though rare, aspiration of foreign bodies into the lower airway during dental procedures may have sequelae that endanger the patient's health. This article discusses the risks associated with routine dental procedures, patient safety precautions during endodontal therapy, and the medicolegal aspects of professional liability in dentistry. The case concerns a 31-year-old man who inhaled a dental cutter which impacted in the left posterior basal lobe. Bronchoscopy failed to retrieve the object; a magnetic resonance imaging study was performed and the object was removed by video-assisted thoracic surgery. Procedure-related criticalities, precautionary measures for patient safety, and medicolegal implications of professional negligence were identified from dentistry protocols and a review of the literature. Professional liability hinges on scrupulous exercise of care and adoption of safety precautions even in routine ''low risk'' dental procedures.