From the *Department Operative Unit of “Lymphatic Surgery”; Post Graduate School in Surgery of the Alimentary Tract, Genoa (Confederated with Pisa, Florence, and Siena); Department of General, Specialist & Oncological Surgery, IRCCS University Hospital, San Martino–IST, National Institute for Cancer Research, University School of Medical and Pharmaceutical Sciences; and DISC—Department of Surgical Sciences and Integrated Diagnostics, Section/Center of Research in Lymphology, Lymphatic Surgery & Microsurgery—“Surgical Clinic,” Genoa, Italy; †Neurosurgery Center for Research, Training and Education, Loma Linda University, CA. Received November 4, 2015, and accepted for publication, after revision November 6, 2015. Conflicts of interest and sources of funding: none declared. Reprints: Corradino Campisi, MD, PhD, FACS, Director of the Department Operative Unit of “Lymphatic Surgery”, Coordinator of the Post Graduate School in Surgery of the Alimentary Tract, Genoa, (Confederated with Pisa, Florence, & Siena), Department of General, Specialist & Oncological Surgery, IRCCS University Hospital, San Martino-IST, National Institute for Cancer Research, University School of Medical and Pharmaceutical Sciences, Genoa, DISC—Department of Surgical Sciences and Integrated Diagnostics, Section/Center of Research in Lymphology, Lymphatic Surgery & Microsurgery—“Surgical Clinic,” L.go R.Benzi, 10 – 16132 Genoa, Italy. E-mail: [email protected].
Intact lyophilized nuclei are obtainable from a variety of tissues, either in situ or in culture, by freezing at -156 degrees C, drying at -25 degrees C, and mechanical disassociation in glycerol at 2 degrees C. Centrifugal separation of nuclei is accomplished in an 85 : 15 by volume mixture of glycerol and 3-chloro-1,2 propanediol at 2 degrees C. The method gives homogeneous nuclear preparations in high yield with preservation of labile and water-soluble constituents.
Accumulation of amyloid-β (Aβ) peptide and the hyperphosphorylation of tau protein are major hallmarks of Alzheimer's disease (AD). The causes of AD are not well known but a number of environmental and dietary factors are suggested to increase the risk of developing AD. Additionally, altered metabolism of iron may have a role in the pathogenesis of AD. We have previously demonstrated that cholesterol-enriched diet causes AD-like pathology with iron deposition in rabbit brain. However, the extent to which chelation of iron protects against this pathology has not been determined. In this study, we administered the iron chelator deferiprone in drinking water to rabbits fed with a 2% cholesterol diet for 12 weeks. We found that deferiprone (both at 10 and 50 mg/kg/day) significantly decreased levels of Aβ40 and Aβ42 as well as BACE1, the enzyme that initiates cleavage of amyloid-β protein precursor to yield Aβ. Deferiprone also reduced the cholesterol diet-induced increase in phosphorylation of tau but failed to reduce reactive oxygen species generation. While deferiprone treatment was not associated with any change in brain iron levels, it was associated with a significant reduction in plasma iron and cholesterol levels. These results demonstrate that deferiprone confers important protection against hypercholesterolemia-induced AD pathology but the mechanism(s) may involve reduction in plasma iron and cholesterol levels rather than chelation of brain iron. We propose that adding an antioxidant therapy to deferiprone may be necessary to fully protect against cholesterol-enriched diet-induced AD-like pathology.
Liquid chromatography–tandem mass spectrometry (LC–MS/MS) and multiple reaction monitoring mass spectrometry (MRM-MS) proteomics analyses were performed on eccrine sweat of healthy controls, and the results were compared with those from individuals diagnosed with schizophrenia (SZ). This is the first large scale study of the sweat proteome. First, we performed LC–MS/MS on pooled SZ samples and pooled control samples for global proteomics analysis. Results revealed a high abundance of diverse proteins and peptides in eccrine sweat. Most of the proteins identified from sweat samples were found to be different than the most abundant proteins from serum, which indicates that eccrine sweat is not simply a plasma transudate and may thereby be a source of unique disease-associated biomolecules. A second independent set of patient and control sweat samples were analyzed by LC–MS/MS and spectral counting to determine qualitative protein differential abundances between the control and disease groups. Differential abundances of selected proteins, initially determined by spectral counting, were verified by MRM-MS analyses. Seventeen proteins showed a differential abundance of approximately 2-fold or greater between the SZ pooled sample and the control pooled sample. This study demonstrates the utility of LC–MS/MS and MRM-MS as a viable strategy for the discovery and verification of potential sweat protein disease biomarkers.
Quantitative changes in the substrates of glycolysis and other metabolites were measured in a series of completely ischemic human and experimentally induced intracranial tumors. Serial portions of the same tumor were maintained under nitrogen at 37°C and then rapidly frozen. Initial rates of lactate production by all tumors were slower than in normal brain, but the final amounts produced at equilibrium were greater. In each tumor, the total amount of lactate produced significantly exceeded the disappearance of glucose and glycogen, and in most cases the increases in Pi exceeded the disappearance of adenosine triphosphate (ATP), P-creatine, and measured phosphorylated intermediates. The source or sources of the extra Pi and lactate was not determined. ATP was maintained at substantial levels despite prolonged ischemia except in 1 tumor, an oligodendroglioma. The significance of these observations in relation to the survival of the neoplastic cell is discussed.
A case is presented in which intraoperative visual evoked response (VER) monitoring was employed during correction of orbital hypertelorism. This procedure is noninvasive and does not interfere with the execution of the operation. The operative time is prolonged only a few minutes to record the VER. This technique is a simple and safe method for detecting intraoperative damage to the optic nerve and chiasm, and may prove useful in avoiding damage to these critical structures during craniofacial surgical procedures.
Background and Objective: The ideal hemostatic agent for laparoscopic partial nephrectomy (LPN) would provide complete hemostasis and sealing of the collecting system at a low cost. Chitosan (CS) is an established topical hemostatic agent, but standard sterilization techniques affect its functional and biologic properties, thereby preventing parenteral uses. This study sought to characterize the safety and efficacy of an implanted CS hemostat sterilized with either a standard technique, electron beam (e-beam) irradiation, or a novel technique, nonthermal nitrogen plasma, in a porcine LPN model. Methods: Laparoscopic partial nephrectomies were performed on six farm pigs and hemostasis achieved using only a CS hemostatic agent (Clo-Sur P.A.D.) that was e-beam (n = 3) or plasma sterilized (PS) (n = 3). Number of pads needed to achieve hemostasis, estimated blood loss, operative time, mass of kidney resection, and warm ischemia time were measured. Animals were monitored for 14 weeks and at harvest, retrograde ureteropyelography and histologic analysis were performed. Results: Complete hemostasis and collection system sealing were achieved in both groups. There was a trend toward less pads required for hemostasis (p = 0.056) and reduced blood loss (p = 0.096) with PS pads, although this did not achieve statistical significance. No complications were observed for 14 weeks and gross examination showed the implanted CS was encapsulated in a fibrous capsule. Histologic analysis revealed a healed nephrectomy site with residual CS and associated chronic inflammation, reactive fibrosis, and foreign body giant cell formation. Importantly, the adjacent renal tissue was intact and viable with no residual parenchymal inflammation or cytologic damage. Conclusion: CS pads alone provided safe and effective hemostasis in a porcine LPN model. PS may enhance hemostatic efficacy and resorption compared with e-beam.
