Circulating Biomarkers of Endothelial Dysfunction and Inflammation in Predicting Clinical Outcomes in Diabetic Patients with Critical Limb Ischemia

Critical limb ischemia (CLI) is a severe manifestation of peripheral artery disease characterized by ischemic pain, which is frequently associated with diabetes and non-healing lesions to inferior limbs. The clinical management of diabetic patients with CLI typically includes percutaneous transluminal angioplasty (PTA) to restore limb circulation and surgical treatment of diabetic foot ulcers (DFU). However, even after successful treatment, CLI patients are prone to post-procedure complications, which may lead to unplanned revascularization or foot surgery. Unfortunately, the factors predicting adverse events in treated CLI patients are only partially known. This study aimed to identify potential biomarkers that predict the disease course in diabetic patients with CLI. For this purpose, we measured the circulating levels of a panel of 23 molecules related to inflammation, endothelial dysfunction, platelet activation, and thrombophilia in 92 patients with CLI and DFU requiring PTA and foot surgery. We investigated whether these putative biomarkers were associated with the following clinical endpoints: (1) healing of the treated DFUs; (2) need for new revascularization of the limb; (3) appearance of new lesions or relapses after successful healing. We found that sICAM-1 and endothelin-1 are inversely associated with DFU healing and that PAI-1 and endothelin-1 are associated with the need for new revascularization. Moreover, we found that the levels of thrombomodulin and sCD40L are associated with new lesions or recurrence, and we show that the levels of these biomarkers could be used in a decision tree to assign patients to clusters with different risks of developing new lesions or recurrences.     

Bacteriology of unshelled frozen blue swimming crab (Portunus pelagicus)

In this study, 45 (10 whole specimens and 35 frozen claws) frozen samples of Portunus pelagicus imported into Sicily (Italy) from the west coast of Africa were examined to assess their bacteriological characteristics and suitability for consumption. Bacteriological examination was performed on two subsamples for each whole crab. The first was the body and claw muscle; the second was a pool of viscera and gills. In the case of frozen claws, each muscle claw was a sample. An aerobic plate count at 30 degrees C (mesophilic aerobic plate count [MAPC]) and 18 degrees C (psychrotrophic aerobic plate count [PAPC]) for 3 days, sulfite-reducing anaerobes, total coliforms, Escherichia coli, enterococci, and Aeromonas spp. were enumerated. Detection of halophilic Vibrio spp. was also performed using salt polymixin broth as an enrichment medium and thiosulfate citrate bile salts sucrose agar as a selective medium; a further morphological and biochemical identification of suspected colonies was performed. The bacterial load of muscle and viscera and gills was low. The MAPC ranged from 0.78 to 3.26 log CFU/g, and the PAPC ranged from 0.48 to 2.41 log CFU/g. Vibrio spp., Aeromonas spp., sulfite-reducing anaerobes, and E. coli were never isolated from muscles or viscera and gills. In contrast to the findings of others, this study showed good bacteriological quality of crabs imported into Sicily from the west coast of Africa. This study also demonstrated the positive influence of the characteristics of environment of origin and postharvest handling hygiene; these parameters could be useful in the context of the application of the hazard analysis critical control point system to this production.     

Praseodymium based high-k dielectrics grown on Si and SiC substrates

High-k polycrystalline Pr₂O₃ thin films have been deposited by metal organic chemical vapor deposition (MOCVD) technique on Si(0 0 1) and 4H–SiC(0 0 0 1) substrates. MOCVD processes have been carried out from the Pr(tmhd)₃ (H-tmhd= 2,2,6,6-tetramethyl-3,5-heptandione) precursor. Complete structural and morphological characterization of films has been carried out using several techniques (X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM)). Polycrystalline Pr₂O₃ films have been obtained and at the interface a praseodymium silicate amorphous layer has been observed on both substrates. The electrical properties of the dielectric praseodymium films have been evaluated.     

Prevalence and distribution of Sarcocystis spp. cysts in several muscles of cattle slaughtered in Sicily,Southern Italy

The aim of this study was to analyse the Sarcocystis spp. infection rate in 22 muscles of cattle, also assessing the diagnostic performance (i.e., sensitivity and concordance) of some non-target muscles. Sarcocysts were detected in the 96% of animals. The 48% of subjects had from 6 to 12 infected muscles while only the 6% had up to 21 positive muscles. The highest rate of infection, 74%, was observed in the heart but the sensitivity registered for some non-target muscle was also acceptable, as in the case of the musculus urethralis with a sensitivity of 59.5%. For this reason, if this site (containing the disseminated prostate) is used for the histological screening of forbidden hormonal treatment, it can allow to find the parasitosis improving the related surveillance activity.     

A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions

Mesenchymal stem cells (MSCs) are classified as advanced therapy medicinal products, a new category of GMP (good manufacturing practice)-compliant medicines for clinical use. We isolated MSCs from 5 bone marrow (BM) samples using human platelet lysate (HPL) instead of foetal bovine serum (FBS). We used a new method of HPL production consisting of treating platelet (PLTs) pools with Ca-Gluconate to form a gel clot, then mechanically squeezing to release growth factors. We compared the new HPL (HPL-S) with the standard (HPL-E) obtained by freezing/thawing cycles and by adding heparin. HPL-S had not PLTs and fibrinogen but the quantity of proteins and growth factors was comparable to HPL-E. Therefore, HPL-S needed fewer production steps to be in compliance with GMP conditions. The number of colonies forming unit-fibroblasts (CFU-F) and the maintenance of stem markers showed no significant differences between MSCs with HPL-E and HPL-S. The cumulative population doubling was higher in MSCs with HPL-E in the earlier passages, but we observed an inverted trend of cell growth at the fourth passage. Immunophenotypic analysis showed a significant lower expression of HLA-DR in the MSCs with HPL-S (1.30%) than HPL-E (14.10%). In conclusion, we demonstrated that HPL-S is an effective alternative for MSC production under GMP conditions.