GC-MS Determination of Flunitrazepam and its Major Metabolite in Whole Blood and Plasma

A gas chromatography-mass spectrometry method was developed for the analysis of flunitrazepam (FN) and its major metabolite, 7-amino-flunitrazepam (7-amino-FN), in both plasma and whole blood. The method was based on acid hydrolysis of the samples after dilution with HPLC water followed by extraction and derivatization (heptafluorobutyrate) of the resulting benzophenones. Analysis of plasma and whole blood samples from subjects administered 2-mg doses of FN showed that FN was only detected in whole blood (LOD 5 ng/mL) and not in plasma. However, 7-Amino-FN was detected in both plasma and whole blood, although the levels were much higher in plasma. 7-Amino-FN was detected for the entire period of specimen collection (12 h), but FN was only detected in whole blood for 4 h after ingestion with peak levels after 1

Evaluation of an Internet Document Delivery Service

An Internet-based Document Delivery Service (DDS) has been developed within the framework of the CNR ( the Italian Research National Council) Project BiblioMIME, in order to take advantage of new Internet technologies and promote cooperation among CNR and Italian university libraries. Adopting such technologies changes the traditional organisation of DDS and may drastically reduce costs and delivery times. An information system managing DDS requests and monitoring the temporal evolution of the service has been implemented, running on the local-area network of a test-site library. It aims to track number and types of documents requested and received, user distribution, delivery times and types (surface mail, fax, Internet), to automate repetitive manual procedures and to deal with the various accounting methods used by other libraries. Transmission of documents is carried out by means of an e-mail/Web gateway system supporting document exchange via Internet, which assists receiving libraries in retrieving requested documents. This paper describes the architecture and main design features of the e-mail/Web gateway server (the BiblioMime server). This approach permits librarians to continue using e-mail service to send large documents, while resolving problems that users may encounter when downloading large size files with e-mail agents. The library operator sends the document as an attachment to the destination address; on fly the e-mail server extracts and saves the attachments in a web-server disk file and substitutes them with a new message part that includes an URL pointing to the saved document. The receiver can download these large objects by means of a user-friendly browser. We further discuss the data gathered during the triennium 1998-2000; this consists of about 5,000 DDS transactions per annum with 300 other Italian scientific and bio-medical libraries and commercial document suppliers. Use of the instruments described above allowed us to evaluate the performance of service "before" and "after" the use of Internet Document Delivery and to extract some critical data regarding DDS. Those include: a) libraries with which we have greater numbers of exchanges and their turnaround times; b) extraordinary reduction in costs and delivery times; c) the most frequently requested serial titles (allowing cost-effective decisions on new subscriptions); d) impact on DDS of library participation in consortia which allow user access to greater numbers of online serials

Mimicking Molecular Pathways in the Design of Smart Hydrogels for the Design of Vascularized Engineered Tissues

Biomaterials are pivotal in supporting and guiding vascularization for therapeutic applications. To design effective, bioactive biomaterials, understanding the cellular and molecular processes involved in angiogenesis and vasculogenesis is crucial. Biomaterial platforms can replicate the interactions between cells, the ECM, and the signaling molecules that trigger blood vessel formation. Hydrogels, with their soft and hydrated properties resembling natural tissues, are widely utilized; particularly synthetic hydrogels, known for their bio-inertness and precise control over cell-material interactions, are utilized. Naturally derived and synthetic hydrogel bases are tailored with specific mechanical properties, controlled for biodegradation, and enhanced for cell adhesion, appropriate biochemical signaling, and architectural features that facilitate the assembly and tubulogenesis of vascular cells. This comprehensive review showcases the latest advancements in hydrogel materials and innovative design modifications aimed at effectively guiding and supporting vascularization processes. Furthermore, by leveraging this knowledge, researchers can advance biomaterial design, which will enable precise support and guidance of vascularization processes and ultimately enhance tissue functionality and therapeutic outcomes

3d collagen hydrogel promotes in vitro langerhans islets vascularization through ad-mvfs angiogenic activity

Adipose derived microvascular fragments (ad-MVFs) consist of effective vascularization units able to reassemble into efficient microvascular networks. Because of their content in stem cells and related angiogenic activity, ad-MVFs represent an interesting tool for applications in regenerative medicine. Here we show that gentle dissociation of rat adipose tissue provides a mixture of ad-MVFs with a length distribution ranging from 33–955 µm that are able to maintain their original morphology. The isolated units of ad-MVFs that resulted were able to activate transcriptional switching toward angiogenesis, forming tubes, branches, and entire capillary networks when cultured in 3D collagen type-I hydrogel. The proper involvement of metalloproteases (MMP2/MMP9) and serine proteases in basal lamina and extracellular matrix ECM degradation during the angiogenesis were concurrently assessed by the evaluation of alpha-smooth muscle actin (αSMA) expression. These results suggest that collagen type-I hydrogel provides an adequate 3D environment supporting the activation of the vascularization process. As a proof of concept, we exploited 3D collagen hydrogel for the setting of ad-MVF–islet of Langerhans coculture to improve the islets vascularization. Our results suggest potential employment of the proposed in vitro system for regenerative medicine applications, such as the improving of the islet of Langerhans engraftment before transplantation

Integrated smart system for energy audit: methodology and application

Abstract The article describes the design and the application stage of a smart energy audit system, integrated within building, and the methodologies adopted for the detection of malfunctions of the plant. The system is set up as a "black box" consisting of a hardware aimed at logging both energy and environmental parameters and a software for the assessment of building behavior and the management of energy flows. The Energy Signature was chosen as the reference method for the evaluation of the energy performance of building. The system was tested in an existing public office building

Battery lifetime of electric vehicles by novel rainflow-counting algorithm with temperature and C-rate dynamics: Effects of fast charging, user habits, vehicle-to-grid and climate zones

The adoption of electric vehicles is expected to soon widespread to cope with energy transition needs; however, concerns on battery lifetime arise, especially related to charging behaviors, vehicle usage habits, vehicle-to -grid and weather conditions. In fact, lifetime battery modeling is a challenging dynamic to characterize, as it involves complex chemical processes related to charging, discharging and temperature dynamics over long time spans that are often difficult to dominate, given the large uncertainties. Having a fatigue-like behavior, the battery aging has sometimes been modeled using rainflow-counting algorithms, yet traditional modeling is not holistic and approximations are used, especially when considering temperature or current dynamics. Based on experimental data, this paper aims at developing a holistic battery degradation model based on rainflow-counting algorithm to properly account for all major determinants of capacity loss, namely cycling usage, calendar lifetime, dynamic temperature and battery current. The approach is coupled with a physical-electro-thermal modeling of the vehicle system, developed in Modelica language, to accurately simulate the intertwined thermal and electrical behavior of the system subject to different usage charging behaviors, including slow and fast charging, as well as vehicle-to-grid application. The proposed case study shows the expected lifetime of electric vehicles to be comparable with of traditional cars (10-20y) and that the proposed temperature -dependent battery modeling enables reducing estimation errors up to 27%. A sensitivity on different climate zones has been considered and results suggest that cool climates can increase life expectancy by 30% with respect to hot climates in typical Italian contexts

Neural Crest-Derived Chondrocytes Isolation for Tissue Engineering in Regenerative Medicine

Chondrocyte transplantation has been successfully tested and proposed as a clinical procedure aiming to repair articular cartilage defects. However, the isolation of chondrocytes and the optimization of the enzymatic digestion process, as well as their successful in vitro expansion, remain the main challenges in cartilage tissue engineering. In order to address these issues, we investigated the performance of recombinant collagenases in tissue dissociation assays with the aim of isolating chondrocytes from bovine nasal cartilage in order to establish the optimal enzyme blend to ensure the best outcomes of the overall procedure. We show, for the first time, that collagenase H activity alone is required for effective cartilage digestion, resulting in an improvement in the yield of viable cells. The extracted chondrocytes proved able to grow and activate differentiation/dedifferentiation programs, as assessed by morphological and gene expression analyses

Identification of phenolic compounds from inflorescences of non-psychoactive Cannabis sativa L. by UHPLC-HRMS and in vitro assessment of the antiproliferative activity against colorectal cancer

Phenolic compounds from Cannabis sativa L. (Cannabaceae family), in particular cannflavins, are known to possess several biological properties. However, their antiproliferative activity, being of great interest from a medicinal chemistry point of view, has not been deeply investigated so far in the literature. In the light of this, the aim of this study was to obtain an enriched fraction of polyphenols (namely PEF) from inflorescences of a non-psychoactive C. sativa (hemp) variety and to evaluate its antiproliferative activity against cancer cells, capitalizing on a new and selective extraction method for hemp polyphenols, followed by preparative flash column chromatography. Untargeted metabolomics, using a new method based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS), was applied here for the first time to fully characterize PEF. Then, the main phenolic compounds were quantified by HPLC-UV. The antiproliferative activity of PEF and of the isolated compounds was assessed in vitro for the first time against Caco-2 and SW480 human colon adenocarcinoma cell lines providing promising IC50 values, in comparison with the reference drug used in therapy for this cancer type. Based on these results, PEF can be considered as a new highly potential therapeutic product to be further investigated against colorectal cancer, thanks to the possible synergistic interaction of its compounds

Mannheim Peritonitis Index (MPI) and elderly population: Prognostic evaluation in acute secondary peritonitis

Introduction. Acute Secondary Peritonitis due to abdominal visceral perforation is characterized by high mortality and morbidity risk. Risk stratification allows prognosis prediction to adopt the best surgical treatment and clinical care support therapy. In Western countries elderly people represent a significant percentage of population Aim. Evaluation of Mannheim Peritonitis Index (MPI) and consideration upon old people. Patients and methods. Retrospective study on 104 patients admitted and operated for \ue2\u80\u9cAcute Secondary Peritonitis due to visceral perforation\ue2\u80\u9d. MPI was scored. In our study we want to demonstrate efficacy of MPI and the possibility to consider older age an independent prognostic factor. Results. Mortality was 25.96%. Greatest sensitivity and specificity for the MPI score as a predictor of mortality was at the score of 20. MPI score of 22. Patients with MPI score 17-21 had 0.46 times lower risk of mortality compared to patients with MPI score >21. In the group of patients with MPI score of >20 the mortality rate was 48.5% for patients older than 80 years old and 12.1% for younger patients (p < 0.005); in the group with MPI score of < 20 mortality rate was respectively 8.4% and 1.4% (p < 0.005). Discussion and conclusions. Data confirm the accuracy of the test. MPI score and age over 80 years old resulted independent predictors of mortality at multivariate analysis