Matrine and oxymatrine in corroborant plant extracts and fertilizers: HPLC/MS-MS method development and single-laboratory validation.

A reversed phase high-performance liquid chromatographic method (HPLC/MS-MS) has been developed and validated for detection of alkaloids matrine and oxymatrine in fertilizer with labeled enhancer plant defense activities. The analytical method was validated statistically. The results show a strong matrix effect, requiring quantification by standard addition method. The regression lines showed r(2) > 0.994. Recoveries ranging from 97 to 104% were obtained for the fortification level of 0.01% wt wt(-1) and the relative standard deviations ranged from 3 to 4% (n = 10). The limits of detection were below 0.0001% wt wt(-1), while the limits of quantification did not exceed 0.0004% wt wt(-1). The method is currently applied in ICQRF Laboratory of Catania on fertilized and corroborant plant extract collected in the Italian market in the frame of MIPAAF institutional quality control activity, with the aim to dectect these unpermitted active substances.

Antiangiogenic VEGF isoform in inflammatory myopathies.

OBJECTIVE: To investigate expression of vascular endothelial growth factor (VEGF) antiangiogenic isoform A-165b on human muscle in idiopathic inflammatory myopathies (IIM) and to compare distribution of angiogenic/antiangiogenic VEGFs, as isoforms shifts are described in other autoimmune disorders. SUBJECTS AND METHODS: We analyzed VEGF-A165b and VEGF-A by western blot and immunohistochemistry on skeletal muscle biopsies from 21 patients affected with IIM (polymyositis, dermatomyositis, and inclusion body myositis) and 6 control muscle samples. TGF- ß, a prominent VEGF inductor, was analogously evaluated. Intergroup differences of western blot bands density were statistically examined. Endomysial vascularization, inflammatory score, and muscle regeneration, as pathological parameters of IIM, were quantitatively determined and their levels were confronted with VEGF expression. RESULTS: VEGF-A165b was significantly upregulated in IIM, as well as TGF- ß. VEGF-A was diffusely expressed on unaffected myofibers, whereas regenerating/atrophic myofibres strongly reacted for both VEGF-A isoforms. Most inflammatory cells and endomysial vessels expressed both isoforms. VEGF-A165b levels were in positive correlation to inflammatory score, endomysial vascularization, and TGF- ß. CONCLUSIONS: Our findings indicate skeletal muscle expression of antiangiogenic VEGF-A165b and preferential upregulation in IIM, suggesting that modulation of VEGF-A isoforms may occur in myositides.

[Thyroidectomy using the intra-operative neuromonitoring and the surgeon's confidence]. / Tiroidectomia totale con neuromonitoring. Esperienza sulla compliance dei chirurghi.

AIM: Reporting our experience with NIM-response 2.0 and 3.0 system in a series of 127 thyroidectomies. MATERIALS OF STUDY: 127 patients were operated at the "Magna Graecia" University (Catanzaro) from 2009 September to 2010 December, using the technique of IntraOperative NeuroMonitoring (IONM). For each patient the surgeon charge filled in a "questionnaire" assessing his/her compliance with the apparatus. RESULTS: We report 0.8% permanent and 0.4% transient recurrent laryngeal nerve paralysis. All surgeons have been fully satisfied from IONM technique. DISCUSSION: A recurrent laryngeal nerve lesion may occur in patients operated on total thyroidectomy between 5-8% for transient palsies and 1-3% for the permanent ones. The use of IONM seems to contribute to a reduction of transient paralysis, even if this method is still not widely accepted Nevertheless the compliance of the surgeons with IONM seems to develop very highly. CONCLUSIONS: NIM-response is helpful in protecting recurrent laryngeal nerve function. Advantages were observed in reduction of postoperative dysphonia and in improving the surgeon's confidence in performing a total thyroidectomy.

Safety and efficacy of continuous or intermittent enteral nutrition in patients in the intensive care unit: Systematic review of clinical evidence.

The best mode of delivering enteral nutrition (EN) in the intensive care unit (ICU) is still debated: several consensus guidelines (American Society for Parenteral and Enteral Nutrition [ASPEN] and the European Society for Parental and Enteral Nutrition [ESPEN]) suggest that EN in ICU should be preferably delivered continuously rather intermittently, but some authors highlight that the first is unphysiological. The aim of this systematic review (SR) is to summarize available clinical evidence related to safety and efficacy of continuous EN (C-EN) or intermittent EN (I-EN) in patients in the ICU, in relation to appropriated supply on nutrition status, gastrointestinal symptoms or tolerance, and risks on respiratory tract infections. A literature search of PubMed, EMBASE, and Google Scholar was performed comparing C-EN vs I-EN, and 4196 published studies were screened. Nineteen studies were selected for this SR reporting types of ICU, nutrition protocols, and study period. Effects of C-EN vs I-EN were presented according to the impact on nutrition status, digestive tract, and respiratory tract. The contrasting results confirmed that the optimal delivering mode of EN remains controversial. Future studies dedicated to identifying the benefits and limitations of C-EN or I-EN should be realized.

Use of CPAP Failure Score to Predict the Risk of Helmet-CPAP Support Failure in COVID-19 Patients: A Retrospective Study.

(1) Background: the aim of this study was to create a score to predict the incidence of CPAP failure in COVID-19 patients early. (2) Methods: in this retrospective observational study, we included all consecutive adult patients admitted between February and April 2021. The main outcome was the failure of CPAP support (intubation or death). (3) Results: two-hundred and sixty-three COVID-19 patients were managed with CPAP. The population was divided in short-CPAP (CPAP days ≤ 10; 72.6%) and long-CPAP (>10; 27.4%) groups. After balancing the entire population using a stabilized IPTW method, we applied a multivariable logistic regression analysis to identify the risk factors for CPAP failure. We used the identified covariates to create a mathematical model, the CPAP Failure Score (CPAP-FS). The multivariable logistic regression analysis identified four variables: SpO2 (OR = 0.86; p-value = 0.001), P/F ratio (OR = 0.99; p-value = 0.008), the Call Score (OR = 1.44; p-value = 0.02), and a pre-existing chronic lung disease (OR = 3.08; p-value = 0.057). The beta-coefficients obtained were used to develop the CPAP-FS, whose diagnostic ability outperformed other relevant COVID-19-related parameters (AUC = 0.87; p-value < 0.0001). We validated the CPAP-FS using a 10-fold internal cross-validation method which confirmed the observed results (AUCs 0.76−0.80; p-values < 0.0001). (4) Conclusions: the CPAP-FS can early identify COVID-19 patients who are at risk of CPAP failure.

Electrospun PLGA Fiber Diameter and Alignment of Tendon Biomimetic Fleece Potentiate Tenogenic Differentiation and Immunomodulatory Function of Amniotic Epithelial Stem Cells.

Injured tendons are challenging in their regeneration; thus, tissue engineering represents a promising solution. This research tests the hypothesis that the response of amniotic epithelial stem cells (AECs) can be modulated by fiber diameter size of tendon biomimetic fleeces. Particularly, the effect of electrospun poly(lactide-co-glycolide) (PLGA) fleeces with highly aligned microfibers possessing two different diameter sizes (1.27 and 2.5 µm: ha1- and ha2-PLGA, respectively) was tested on the ability of AECs to differentiate towards the tenogenic lineage by analyzing tendon related markers (Collagen type I: COL1 protein and mRNA Scleraxis: SCX, Tenomodulin: TNMD and COL1 gene expressions) and to modulate their immunomodulatory properties by investigating the pro- (IL-6 and IL-12) and anti- (IL-4 and IL-10) inflammatory cytokines. It was observed that fiber alignment and not fiber size influenced cell morphology determining the morphological change of AECs from cuboidal to fusiform tenocyte-like shape. Instead, fleece mechanical properties, cell proliferation, tenogenic differentiation, and immunomodulation were regulated by changing the ha-PLGA microfiber diameter size. Specifically, higher DNA quantity and better penetration within the fleece were found on ha2-PLGA, while ha1-PLGA fleeces with small fiber diameter size had better mechanical features and were more effective on AECs trans-differentiation towards the tenogenic lineage by significantly translating more efficiently SCX into the downstream effector TNMD. Moreover, the fiber diameter of 1.27 µm induced higher expression of pro-regenerative, anti-inflammatory interleukins mRNA expression (IL-4 and IL-10) with favorable IL-12/IL-10 ratio with respect to the fiber diameter of 2.5 µm. The obtained results demonstrate that fiber diameter is a key factor to be considered when designing tendon biomimetic fleece for tissue repair and provide new insights into the importance of controlling matrix parameters in enhancing cell differentiation and immunomodulation either for the cells functionalized within or for the transplanted host tissue.

Tendon Biomimetic Electrospun PLGA Fleeces Induce an Early Epithelial-Mesenchymal Transition and Tenogenic Differentiation on Amniotic Epithelial Stem Cells.

Background. The design of tendon biomimetic electrospun fleece with Amniotic Epithelial Stem Cells (AECs) that have shown a high tenogenic attitude may represent an alternative strategy to overcome the unsatisfactory results of conventional treatments in tendon regeneration. Methods. In this study, we evaluated AEC-engineered electrospun poly(lactide-co-glycolide) (PLGA) fleeces with highly aligned fibers (ha-PLGA) that mimic tendon extracellular matrix, their biocompatibility, and differentiation towards the tenogenic lineage. PLGA fleeces with randomly distributed fibers (rd-PLGA) were generated as control. Results. Optimal cell infiltration and biocompatibility with both PLGA fleeces were shown. However, only ha-PLGA fleeces committed AECs towards an Epithelial-Mesenchymal Transition (EMT) after 48 h culture, inducing their cellular elongation along the fibers' axis and the upregulation of mesenchymal markers. AECs further differentiated towards tenogenic lineage as confirmed by the up-regulation of tendon-related genes and Collagen Type 1 (COL1) protein expression that, after 28 days culture, appeared extracellularly distributed along the direction of ha-PLGA fibers. Moreover, long-term co-cultures of AEC-ha-PLGA bio-hybrids with fetal tendon explants significantly accelerated of half time AEC tenogenic differentiation compared to ha-PLGA fleeces cultured only with AECs. Conclusions. The fabricated tendon biomimetic ha-PLGA fleeces induce AEC tenogenesis through an early EMT, providing a potential tendon substitute for tendon engineering research.

Dehydroacetic acid in cheese and cheese coating, results of official control in Italy.

This paper reports the occurrence of dehydroacetic acid in cheese and cheese coatings collected in Italy during Agricultural Ministry Official control. Dehydroacetic acid is an antimicrobial substance not allowed to be used in EU countries as a food additive, with unknown effects on human health. Dehydroacetic acid was measured by a validated HPLC method according to Commission Decision 2002/657/EC criteria in terms of specificity, linearity, precision and accuracy, limit of detection, and limit of quantification. The method was successfully applied to 129 samples of commercial cheese coatings and related treated cheeses collected in Italy during 2017. The overall results demonstrated that about 40% of the investigated cheese coatings contained dehydroacetic acid, ranging from 0.010% to 2.5% w/w, evidencing illicit employment of this substance. Moreover, about 25% of treated cheeses contained dehydroacetic acid, from 5 to 250 mg/Kg, proving transfer of this substance from crust to cheese.

Platinum-based antitumor drugs containing enantiomerically pure alpha-trifluoromethyl alanine as ligand.

Synthetic amino acids such as fluorinated alpha-amino acids are currently actively investigated for their biological activity. Herein, we report on the synthesis and characterization of platinum complexes containing an N,O-chelated pure enantiomer of alpha-trifluoromethylalanine (alpha-Tfm-Ala). The compounds are either anionic, K[PtCl2(alpha-Tfm-Ala)], or cationic, [PtAm2(alpha-Tfm-Ala)](NO3) (Am2= (NH3)2, ethylendiamine (en), 1,10-phenanthroline (phen), and 2,9-dimethyl-1,10-phenanthroline (Me2phen)). All complexes are highly soluble in water and have been fully characterized by NMR spectroscopy. In vitro cytotoxicity assays on different human tumor cell lines have been performed on some of the isolated compounds. [Pt(NH3)2(alpha-Tfm-Ala)] with R configuration of the amino acid proved to have an activity comparable to that of the reference compound cisplatin. Flow cytometric analysis on NCI-H460 tumor cells (absence of G2/M arrest, which instead is observed in the case of cisplatin) suggests a mechanism of action different from that of cisplatin.

Aminocarminic acid in E120-labelled food additives and beverages.

An analytical method was developed for investigating aminocarminic acid occurrence in E120-labelled red-coloured-beverages and in E120 additives, with the aim of controlling the purity of the carmine additive in countries where the use of aminocarminic acid is forbidden. The carminic acid and the aminocarminic acid were separated by high-performance liquid chromatography-photodiode array-tandem mass spectrography (HPLC-PDA-MS/MS). The method was statistically validated. The regression lines, ranging from 10 to 100 mg/L, showed r(2 )> 0.9996. Recoveries from 97% to 101% were obtained for the fortification level of 50 mg/L; the relative standard deviations did not exceed 3%. The LODs were below 2 mg/L, whereas the LOQs did not exceed 4 mg/L. The method was successfully applied to 27 samples of commercial E120-labelled red-coloured beverages and E120 additives, collected in Italy during quality control investigations conducted by the Ministry. The results demonstrated that more than 50% of the samples contained aminocarminic acid, evidencing the alarming illicit use of this semi-synthetic carmine acid derivative.

Solid-supported solution-phase synthesis of 4-amino-1,2,4,5-tetrahydro-2-benzazepine-3-ones.

Starting from Boc-o-aminomethylphenylalanine, a solution-phase parallel synthesis of 2,4-substituted 4-amino-1,2,4,5-tetrahydro-2-benzazepine-3-ones is described. This heterocycle has two nitrogen functions, which are differentiated and can be selectively substituted. The sources of diversity are aldehydes for the R(1) position and carboxylic acids, sulfonyl chlorides, or isocyanates for the R(2) position. High-throughput synthesis and purification of this multistep synthetic sequence was accomplished using polymer-bound reagents and scavengers and liquid-liquid extraction protocols, and a small library of compounds was prepared. Polymer-bound cyanoborohydride was found to work well for the reductive amination. Scavenging of excess of amine was performed by polymer-bound benzaldehyde, and cyclization was performed in the presence of polymer-bound coupling reagent 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC). After Boc-deprotection, the second nitrogen can be acylated using carboxylic acids, sulfonylated or converted to a urea. The acylation is again performed by polymer-bound EDC. Excellent yields and purities were obtained.