Development of an analytical platform for the affinity screening of natural extracts by SEC-MS towards PPARα and PPARγ receptors.

BACKGROUND: Peroxisome proliferator-activated receptors (PPARs) belong to the superfamily of nuclear receptors and represent the targets for the therapeutical treatment of type 2 diabetes, dyslipidemia and hyperglycemia associated with metabolic syndrome. Some medicinal plants have been traditionally used to treat this kind of metabolic diseases. Today only few drugs targeting PPARs have been approved and for this reason, the rapid identification of novel ligands and/or chemical scaffolds starting from natural extracts would benefit of a selective affinity ligand fishing assay. RESULTS: In this paper we describe the development of a new ligand fishing assay based on size exclusion chromatography (SEC) coupled to LC-MS for the analysis of complex samples such as botanical extracts. The known PPARα and PPARγ ligands, WY-14643 and rosiglitazone respectively, were used for system development and evaluation. The system has found application on an Allium lusitanicum methanolic extract, containing saponins, a class of chemical compounds which have attracted interest as PPARs ligands because of their hypolipidemic and insulin-like properties. SIGNIFICANCE: A new SEC-AS-MS method has been developed for the affinity screening of PPARα and PPARγ ligands. The system proved to be highly specific and will be used to improve the throughput for the identification of new selective metabolites from natural souces targeting PPARα and PPARγ.

Candidacy and long-term outcomes of subcutaneous implantable cardioverter-defibrillators in current practice in patients with hypertrophic cardiomyopathy.

BACKGROUND: In patients with Hypertrophic Cardiomyopathy (HCM) S-ICD is usually the preferred option as pacing is generally not indicated. However, limited data are available on its current practice adoption and long-term follow-up. METHODS: Consecutive HCM patients with S-ICD implanted between 2013 and 2021 in 3 international centers were enrolled in this observational study. Baseline, procedural and follow-up data were regularly collected. Efficacy and safety were compared with a cohort of HCM patients implanted with a tv-ICD. RESULTS: Seventy patients (64% males) were implanted with S-ICD at 41 ± 15 years, whereas 168 patients with tv-ICD at 49 ± 16 years. For S-ICD patients, mean ESC SCD risk score was 4,5 ± 1.9%: 25 (40%) at low-risk, 17 (27%) at intermediate and 20 (33%) at high-risk. Patients were followed-up for 5.1 ± 2.3 years. Two patients (0.6 per 100-person-years, vs 0.4 per 100 person-years with tv-ICD, p = 0.45) received an appropriate shock on VF, 17 (24%) were diagnosed with de-novo AF. Inappropriate shocks occurred in 4 patients (1.2 per 100-person-years, vs 0.9 per 100 person-years with tv-ICD, p = 0.74), all before Smart-Pass algorithm implementation. Four patients experienced device-related adverse events (1.2 per 100-person-years, vs 1 per 100 person-years with tv-ICD, p = 0.35%). CONCLUSIONS: S-ICDs were often implanted in patients with an overall low-intermediate ESC SCD risk, reflecting both the inclusion of additional risk markers and a lower decision threshold. S-ICDs in HCM patients followed for over 5 years showed to be effective in conversion of VF and safe. Greater scrutiny may be required to avoid overtreatment in patients with milder risk profiles.

Long-term scarring outcomes and safety of patients treated with NovoSorbⓇ Biodegradable Temporizing Matrix (BTM): An observational cohort study.

Background/Aim: NovoSorbⓇ Biodegradable Temporizing Matrix (BTM) is a relatively novel, biodegradable polyurethane-based dermal regeneration template. The aim of this study was to evaluate the long-term scarring outcomes and safety of BTM in patients who underwent dermal reconstruction involving ≥5% of the total body surface area. Methods: This was a postmarket, multicenter, observational cohort study involving evaluation of long-term outcomes in patients treated with BTM. A total of 55 patients (35 from Royal Adelaide Hospital, South Australia, and 20 from Victoria Adult Burns Service, The Alfred, Victoria) who underwent dermal repair with BTM between 2011 and 2017 were screened for inclusion in this study. All patients had BTM implanted for ≥18 months. Results: Fifteen eligible patients with a mean (SD) age of 49.1 (14.3) years completed study assessments. These patients had a total of 39 areas treated with BTM. Using the Patient and Observer Scar Assessment Scale, scar quality was reported to be good by both observers and patients, with a mean (SD) observer score across all lesions of 3.6 (1.2) and mean (SD) overall opinion of 3.8 (1.2) as well as a mean (SD) patient score of 3.5 (1.2) and overall opinion of 5.0 (2.2). No adverse events or adverse device effects were reported or identified. Conclusion: The long-term scar quality is comparable to published studies. BTM is safe in the long term with no additional risks or adverse consequences being identified.

The Multidisciplinary Approach for the Diagnosis of Laryngohyoid Lesions: a Systematic Literature Review and Meta-Analysis.

Background: The diagnosis of neck lesions remains a medico-legal diagnostic challenge because of the complexity of the anatomical relationship of the neck's organs and their anthropometric morphological variability. We compared the multidisciplinary approach using autopsy and postmortem computed tomography (PMCT), postmortem fine preparation (PMFP), postmortem micro-computed tomography (micro-CT), and postmortem magnetic resonance (PMMR) with the performance of a single diagnostic method among them evaluating the significance of different results. The multidisciplinary approach significantly reduced the number of unidentified neck lesions. The analysis demonstrates the need to better define the scan protocols and compose forensic guidelines for radiological application. The results of this study point out the need to compare the different diagnostic approaches in deceased subjects to better define the radiological scan protocol based on a multidisciplinary approach, including autopsy and radiological methods and the radiological scan protocols. Methods: We performed a systematic electronic search of retrospective scientific articles in PubMed, the Scopus database, and the Cochrane Library. The following combinations of words were used: "hyoid fracture"; "comparison between PMCT AND autopsy"; "hyoid fracture PMCT AND autopsy"; "hyoid bone fracture AND forensic imaging"; "hyoid fracture AND PMCT"; "neck fracture PMCT AND autopsy"; "laryngohyoid lesions"; "postmortem CT AND autopsy in strangulation"; "postmortem AND strangulation Signs "; "strangulation virtopsy"; and "strangulation AND MRI". We selected 16 articles that were published between March 2003 and June 2020. We conducted a meta-analysis with R software to evaluate the rates. We obtained related confidence intervals and a forest plot. Results: Thyroid cartilage damages were significantly more common than hyoid bone fractures (61.7% vs 42.2%) in a sample of 128 subjects. The synergic uses of autopsy/PMCT, autopsy/PMFP, autopsy/microCT, and autopsy/PMMR revealed significantly higher rates than a single investigation. We analyzed the PMCT scan data. The scan parameters evaluated were as follows: row, scan sample, reconstruction, kernel, slice thickness, kVp, and mAs. A lack of uniformity in the application of the protocol was observed. Conclusion: Further studies are needed to better define the radiological scan protocols and to draw guidelines to identify the appropriate radiological methods in relation to the specific case.

Patterns of repeated diagnostic testing for COVID-19 in relation to patient characteristics and outcomes.

BACKGROUND: Whilst the COVID-19 diagnostic test has a high false-negative rate, not everyone initially negative is re-tested. Michigan Medicine, a primary regional centre, provided an ideal setting for studying testing patterns during the first wave of the pandemic. OBJECTIVES: To identify the characteristics of patients who underwent repeated testing for COVID-19 and determine if repeated testing was associated with downstream outcomes amongst positive cases. METHODS: Characteristics, test results, and health outcomes for patients presenting for a COVID-19 diagnostic test were collected. We examined whether patient characteristics differed with repeated testing and estimated a false-negative rate for the test. We then studied repeated testing patterns in patients with severe COVID-19-related outcomes. RESULTS: Patient age, sex, body mass index, neighbourhood poverty levels, pre-existing type 2 diabetes, circulatory, kidney, and liver diseases, and cough, fever/chills, and pain symptoms 14 days prior to a first test were associated with repeated testing. Amongst patients with a positive result, age (OR: 1.17; 95% CI: (1.05, 1.34)) and pre-existing kidney diseases (OR: 2.26; 95% CI: (1.41, 3.68)) remained significant. Hospitalization (OR: 7.88; 95% CI: (5.15, 12.26)) and ICU-level care (OR: 6.93; 95% CI: (4.44, 10.92)) were associated with repeated testing. The estimated false-negative rate was 23.8% (95% CI: (19.5%, 28.5%)). CONCLUSIONS: Whilst most patients were tested once and received a negative result, a meaningful subset underwent multiple rounds of testing. These results shed light on testing patterns and have important implications for understanding the variation of repeated testing results within and between patients.

Barotraumatic blowout fracture of the orbit after sneezing: Cone beam CT demonstration.

A 40-year-old man with no history of trauma or previous sinus surgery complained sudden right diplopia after vigorous sneezing. The patient was submitted elsewhere to a MRI study for persisting diplopia, with inconclusive findings. One week later the patient was submitted to a maxillofacial cone beam CT (CBCT) in our Institution. A blowout fracture of the right orbital floor and lateral orbital wall with an intact orbital rim and a ipsilateral maxillary sinus effusion with an air liquid level, were detected at the CBCT study. Our conclusion, confirmed by the clinician, was the patient had a barotraumatic, isolated, pure blowout fracture of the right orbit consequence of the episode of vigorous sneezing. The patient was treated by steroid and antibiotic therapy and diplopia resolved after two weeks. Clinicians and radiologists should be aware that a barotraumatic blowout fracture of the orbit after sneezing should be included among the unusual causes of sudden diplopia.

Cardiac rupture caused by traffic accident: Case reports and a literature review.

The commonest cause of blunt cardiac injuries is from traffic accidents followed by violent falls, sport activities, accidents or a fight but rupture of the heart is rare and lethal. The precise incidence of cardiac injury after a blunt chest trauma is unknown as rates vary greatly in the literature from between 7% and 76% of cases. Autopsy studies have shown that the right ventricle is the most frequently ruptured, followed by the left ventricle, right atrium, intraventricular septum, left atrium and interatrial septum with decreasing frequency. Post-mortem imaging is a rapidly advancing field of post-mortem investigations of trauma victims. The available literature dealing with the comparison of post-mortem computed tomography results with forensic autopsy indicates that conventional autopsy remains superior for the detection of organ and soft tissue injuries in all body regions.

Utility of post mortem computed tomography in clivus fracture diagnosis. Case illustration and literature review.

Clivus fractures are usually associated with head blunt trauma due to traffic accident and falls. A 23 - year-old man died immediately after a smash-up while he was stopping on his motorcycle. Post-mortem Computed tomography (PMCT), performed before autopsy, revealed a complex basilar skull base fractures associated with brainstem and cranio-vertebral junction injuries, improving the diagnostic performance of conventional autopsy. Imaging data were re-assessable and PMCT offers the possibility to perform multiplanar and volume rendered reconstructions, increasing forensic medicine knowledge related to traumatic injuries.

Prognostic evaluation of biofeedback response in patients treated for anorectal malformation.

PURPOSE: Functional bowel outcome in patients with anorectal malformation often is poor. For fecal incontinence resulting from sphincter dysfunction, biofeedback (BFB) training appears to be effective. The aim of study was to investigate the bowel function in incontinent children treated for ARM, using a clinical score, a manometric and pelvic magnetic resonance evaluation, in order to establish predictive parameters of response after BFB. METHODS: 25 children (median age of 6.5 years) with true fecal incontinence were evaluated by clinical score, anorectal manometry and magnetic resonance imaging (MRI). According to these evaluations patients were divided in 4 groups: group 1 (favorables manometry and MRI); group 2 (favorable manometry and unfavorable MRI); group 3 (unfavorable manometry and favorable MRI); group 4 (unfavorables manometry and MRI). All groups started a cycle of BFB and six months after end of BFB, were reevaluated by clinical score and manometry. RESULTS: The overall response to BFB was excellent in 44%, discrete in 40% and poor in 16%; a better response was found in groups 1 and 2 than groups 3 and 4. The differences between groups before BFB proportionally correlated with values after BFB; a correlation with genitourinary and spinal anomalies was found. CONCLUSIONS: Our results showed that BFB is an effective for fecal incontinence when the assessment pretreatment (functional and morphologic) is favorable; the manometry can evaluate the potential sphincterial recovery after BFB with a further prognostic benefit if correlated to morphologic evaluation with MRI.

[Cell phones: health risks and prevention]. / Telefoni cellulari: rischi per la salute e strategie di prevenzione.

The paper describes first of all the electromagnetic radiation of cellular phones and presents the physical parameters used to measure and evaluate the absorption of emissions of radio stations and cellular phones. It then presents selected research results of the experimental studies in vivo and in vitro which examine the biological effects of the emissions of cellular phones. The review of the epidemiologic evidence focuses in particular the epidemiologic studies on the use of cell phones and brain tumours, identifying some of the reasons of the conflicting results obtained. Studies dealing with the health risks involved in the increasing use of cellular phones by adolescents and children, more sensitive to this exposure, are also presented showing the need for special caution. The problem of hypersensitivity observed in some individuals is also briefly discussed. Finally the paper presents a summary of the main prevention measures necessary in order to reduce the risks in the framework of the "precautionary principle" including prevention policies and exposure limits in various countries.

Geometrically constrained derivatives of indolylglyoxylamides as ligands binding the GABAA/BzR complex.

Indolylglyoxylamides are a class of distinctive benzodiazepine receptor ligands, proposed in the mid-eighties as open analogues of -carbolines. Thorough and long-lasting studies of their structure-activity relationships led to the development of a great deal of derivatives, to satisfy increasingly structural and pharmacophoric requirements of the benzodiazepine binding site in the central nervous system. Efforts to pre-organize their flexible structure in the three-dimensional shape adopted when bound to the receptor led to the identification of two novel classes of rigid ligands, characterized by planar tricyclic heteroaromatic cores: the [1,2,4]triazino[4,3-a]benzimidazol-4(10H)-one and the [1,2,3]triazolo[1,2-a][1,2,4]benzotriazin-1,5(6H)-dione. The present review focuses on these selected classes of ligands, whose rational development, in terms of chemical structures and structure-activity relationships, will be fully discussed.

Pediatric sialolithiasis distinctive characteristic in radiological imaging.

AIM: Aim of the present paper was to investigate the imaging and related clinical characteristics of sialolithiasis in Italian pediatric population trying to determine the difference between pediatric and adult. METHODS: Twenty-nine pediatric patients (age range 1-17 years) with pain and postprandial swelling and/or purulent discharge in the salivary gland areas were referred to radiology department after pediatric ear, nose and throat (ENT) evaluation. They all were submitted to ultrasound examination of the main salivary glands. Multidetector computed tomography (MDCT) only was performed in 2/6 patients, in 2/6 patients both sialography and MDCT were performed due to inconclusive MDCT features, 2/6 only sialography was performed. Sialoliths were classified on their location and size. RESULTS: In 6 out of 29 patients (4 males, 2 females, age range 1-17 years) salivary stones were detected. Sialoliths were detected in 5/6 patients in the submandibular gland and 1/6 in the parotid gland. All sialoliths, excepted for a case of multiple sialoliths, were located in the distal part of the main salivary ducts. CONCLUSION: Imaging characteristics of sialolith in pediatric group are similar than in adult population in few aspects. In fact sialoliths are smaller in size and located more frequently in the distal part of the main salivary duct, than in adult, making sialography cannulation more complex and requiring short thickness in MDCT.

High Resolution CT Angiography in Detection of an Aneurysm of the Vein of Galen as a Source of Intracranial Haemorrhage in a Newborn.

Cerebral haemorrhage is a rare condition in infants and carries a known poor prognosis. Common causes of spontaneous haemorrhage include various vascular venous lesions due to incomplete hydrovenous maturation, among them Galen vein aneurysm may be a very rare cause of cerebral haemorrhage. This report emphasizes the role of multidector CT with high resolution CT angiography in a newborn with cerebral hemorrhage caused by Galen vein aneurysm rupture. MDCT with high resolution CT angiography helps to differentiate the cause of haemorrhage, and to address the appropriate treatment.

[Gender at work: from theory to practice]. / Il genere al lavoro: dalla teoria alla pratica.

INTRODUCTION: In Italy a recent legislation introduced norms concerning gender based risk assessment. METHODS: Collection of recent european and international publication and network Congresses on gender. RESULTS: Italy has now the scientific knowledge to start a gender oriented risk assessment. DISCUSSION: Gender sensitive good practices can represent a good way to improve women working conditions in Italy

Pain assessment in fibromyalgia and rheumatoid arthritis: influence of physical activity and illness perception.

OBJECTIVE: Pain visual analog scales (VAS) have been validated for clinical use in fibromyalgia (FM) and rheumatoid arthritis (RA) patients. There are potential limitations, however, not only considering their use as a continuous measurement, but also with regard to the influence of personal illness perceptions, habitual physical activity and other life-style features. The aim of the study was to ascertain whether different illness perception, physical activity and clinical and laboratory characteristics can predict the severity of perceived pain assessed by VAS. MATERIALS AND METHODS: This is an observational comparative study of forty consecutive out-patients, 20 of them with fibromyalgia and 20 with rheumatoid arthritis, treated by medical and physical therapy. Patients were assessed also by Pain VAS, Health Assessment Questionnaire (HAQ) disability index, Ritchie index, Baecke questionnaire for physical activity, Illness Perception Questionnaire (IPQr) and SF36. RESULTS: Pain VAS is explained differently by some of the studied variables: in the total group HAQ and Ritchie index explain 29.8% of the variance; in the RA patients number of joints with pain and Ritchie index explain 52.7% of the variance; in FM patients total SF36 score and IPQr personal control dimension explains 44.7% of the variance. No definite role of anxiety and/or depression was found as predictor of perceived pain and disability. CONCLUSION: Pain perception and complaint are explained by belief in FM patients: This seems to suggest the need for a more articulated cognitive approach; addressing both diagnostic and therapeutic interventions to anxiety/depression issues is not supported by our results.

Recent advances in the development of dual topoisomerase I and II inhibitors as anticancer drugs.

DNA topoisomerases (topos) are essential enzymes that regulate the topological state of DNA during cellular processes such as replication, transcription, recombination, and chromatin remodeling. Topoisomerase I (Topo I) is a ubiquitous nuclear enzyme which catalyzes the relaxation of superhelical DNA generating a transient single strand nick in the duplex, through cycles of cleavage and religation. Topoisomerase II (Topo II) mediates the ATP-dependent induction of coordinated nicks in both strands of the DNA duplex, followed by crossing of another double strand DNA through the transiently broken duplex. Although the biological functions of Topoisomerases are important for ensuing genomic integrity, the ability to interfere with enzymes or generate enzyme-mediated damage is an effective strategy for cancer therapy and, in this connection, DNA topos (I and II) proved to be the excellent targets of clinically significant classes of anticancer drugs. Actually, specific Topo I and Topo II inhibitors reversibly trap the enzyme-DNA complexes, thus converting Topos into physiological poisons, able to produce permanent DNA damage, which triggers cell death. Given that both enzymes are good targets, it would be desirable to jointly inhibit them, but use-limiting toxicity of sequential or simultaneous combinations of topo I and II poisons include severe to life-threatening neutropenia and anemia. Furthermore, the emergence of resistance phenomena to topo I inhibitors is often accompanied by a concomitant rise in the level of topo II expression and viceversa, leading to the failure of clinical therapies. In this regard, a single compound able to inhibit both Topo I and II may present the advantage of improving antitopoisomerase activity, with reduced toxic side effects, with respect to the combination of two inhibitors. Due to the high interest in such compounds, this review represents an update of previous works dealing with the development of dual Topo I and II inhibitors as novel anti-cancer agents. The newly collected derivatives have been described focusing attention on their chemical structures and their biological profiles.

Sialographic findings in Wharton duct evagination.

Ductal evagination is a rare condition affecting the Wharton duct. The aim of this study was to establish the incidence, imaging features and clinical significance of ductal evagination in patients undergoing submandibular gland sialography. The sialographic findings and reports of 322 patients undergoing submandibular gland sialography during the period 1998-2007 were retrospectively reviewed. Ductal evagination was identified on sialograms as a unique diverticulum, filled with contrast medium, of the Wharton duct, with a narrow neck and a blind end. A ductal evagination was found in 5/322 patients with swelling and pain in the submandibular gland. It was always located in the middle tract of the Wharton duct. Other findings were: in 5/5 patients, an enlargement of the primary and secondary ducts due to sialodochitis; in 3/5 patients, salivary stones; in 1/5 patients, a sinuous distal tract of the Wharton duct; in 2/5 patients, a stenosis of the proximal tract of the Wharton duct. In our series, the incidence of wharton duct evagination was 1.6% incidence. It may represent a form of duct wall weakness although its cause is uncertain. It is, however, a condition that needs to be highlighted on sialograms for eventual inflammatory consequences or in guiding sialoendoscopy to avoid eventual ductal perforation.

Synthesis and biological activity of 1,4-dihydrobenzothiopyrano[4,3-c]pyrazole derivatives, novel pro-apoptotic mitochondrial targeted agents.

This study reports the synthesis of a number of 1- and 2-phenyl derivatives of the 1,4-dihydrobenzothiopyrano[4,3-c]pyrazole nucleus, which were obtained by the reaction of the versatile 7-substituted 2,3-dihydro-3-hydroxymethylene-4H-1-benzothiopyran-4-ones with hydrazine and substituted phenylhydrazines. The antiproliferative activity of the synthesized compounds was evaluated by an in vitro assay on human tumor cell lines (HL-60 and HeLa) and showed a significant capacity of the 7-methoxy-substituted benzothiopyrano[4,3-c]pyrazoles 3b-d, carrying the pendant phenyl group in the 1-position, to inhibit cell growth. Investigation of the mechanism of action indicated the induction of the mitochondrial permeability transition (MPT) as the molecular event responsible for the inhibition of cell growth. This phenomenon is related to the ability of the test compounds to cause a rapid Ca2+-dependent and cyclosporin A-sensitive collapse of the transmembrane potential (DeltaPsi) and matrix swelling. All this leads to the release of caspase activators, such as cytochrome c (cyt c) and apoptosis-inducing factor (AIF), which trigger the pro-apoptotic pathway leading to DNA fragmentation.