Human osteoarthritic chondrocytes exposed to extremely low-frequency electromagnetic fields (ELF) and therapeutic application of musically modulated electromagnetic fields (TAMMEF) systems: a comparative study.

Osteoarthritis (OA) is the most common joint disease, characterized by matrix degradation and changes in chondrocyte morphology and metabolism. Literature reported that electromagnetic fields (EMFs) can produce benefits in OA patients, even if EMFs mechanism of action is debated. Human osteoarthritic chondrocytes isolated from femoral heads were cultured in vitro in bidimensional (2-D) flasks and in three-dimensional (3-D) alginate beads to mimic closely cartilage environment in vivo. Cells were exposed 30 min/day for 2 weeks to extremely low-frequency electromagnetic field (ELF) with fixed frequency (100 Hz) and to therapeutic application of musically modulated electromagnetic field (TAMMEF) with variable frequencies, intensities, and waveforms. Cell viability was measured at days 7 and 14, while healthy-cell density, heavily vacuolized (hv) cell density, and cluster density were measured by light microscopy only for 3-D cultures after treatments. Cell morphology was observed for 2-D and 3-D cultures by transmission electron microscopy (TEM). Chondrocyte exposure to TAMMEF enhances cell viability at days 7 and 14 compared to ELF. Light microscopy analysis showed that TAMMEF enhances healthy-cell density, reduces hv-cell density and clustering, compared to ELF. Furthermore, TEM analysis showed different morphology for 2-D (fibroblast-like) and 3-D (rounded shape) cultures, confirming light microscopy results. In conclusion, EMFs are effective and safe for OA chondrocytes. TAMMEF can positively interfere with OA chondrocytes representing an innovative non-pharmacological approach to treat OA

HISTOPATHOLOGICAL FINDINGS IN SYSTEMIC SCLEROSIS-RELATED MYOPATHY: FIBROSIS AND MICROANGIOPATHY

Objectives: The objective of this study was to identify specific histopathological features of skeletal muscle involvement in systemic sclerosis (SSc) patients. Methods: A total of 35 out of 112 SSc-patients (32%, including 81% female and 68% diffuse scleroderma) presenting clinical, biological and electromyographic (EMG) features of muscle weakness, were included. Patients underwent vastus lateralis biopsy, assessed for individual pathologic features including fibrosis [type I collagen (Coll-I), transforming growth factor β (TGF-β)], microangiopathy [cluster of differentiation 31 (CD31), pro-angiogenic vascular endothelial growth factor A (VEGF-A), anti-angiogenic VEGF-A165b], immune/ inflammatory response [CD4, CD8, CD20, human leucocyte antigens ABC (HLA-ABC)], and membranolytic attack complex (MAC). SSc biopsies were compared with biopsies of (n = 35) idiopathic inflammatory myopathies (IIMs) and to (n = 35) noninflammatory myopathies (NIMs). Ultrastructural abnormalities of SSc myopathy were also analyzed by transmission electron microscopy (TEM). Results: Fibrosis in SSc myopathy (81%) is higher compared with IIM (32%, p < 0.05) and with NIM (18%, p < 0.05). Vascular involvement is dominant in SSc muscle (92%), and in IIM (78%) compared with NIM (21%, p < 0.05). In particular, CD31 shows loss of endomysial vessels in SSc myopathy compared with IIM (p < 0.05) and with NIM (p < 0.01). VEGF-A is downregulated in SSc myopathy compared with IIM (p < 0.05) and NIM (p < 0.05). Conversely, VEGF-A165b is upregulated in SSc myopathy. The SSc immune/inflammatory response suggested humoral process with majority (85%) HLA-ABC fibral neoexpression and complement deposits on endomysial capillaries MAC, compared with IIM (p < 0.05), characterized by CD4+/CD8+/B-cell infiltrate, and NIM (p < 0.05). TEM analysis showed SSc vascular alterations consisting of thickening and lamination of basement membrane and endothelial cell ‘swelling’ coupled to endomysial/perimysial fibrosis. Conclusions: Fibrosis, microangiopathy and humoral immunity are predominant in SSc myopathy, even if it is difficult to identify specific histopathological hallmarks of muscle involvement in SSc, since they could be present also in other (IIM/NIM) myopathies. © 2016, © The Author(s), 2016

Thymoglobulin, interferon-γ and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures

<p>Abstract</p> <p>Background</p> <p>Cytokine-induced killer (CIK) cells are typically differentiated <it>in vitro </it>with interferon (IFN)-γ and αCD3 monoclonal antibodies (mAb), followed by the repeated provision of interleukin (IL)-2. It is presently unknown whether thymoglobulin (TG), a preparation of polyclonal rabbit γ immunoglobulins directed against human thymocytes, can improve the generation efficiency of CIK cells compared with αCD3 mAb in a clinical-grade culture protocol.</p> <p>Methods</p> <p>Peripheral blood mononuclear cells (PBMC) from 10 healthy donors and 4 patients with solid cancer were primed with IFN-γ on day 0 and low (50 ng/ml), intermediate (250 ng/ml) and high (500 ng/ml) concentrations of either αCD3 mAb or TG on day 1, and were fed with IL-2 every 3 days for 21 days. Aliquots of cells were harvested weekly to monitor the expression of representative members of the killer-like immunoglobulin receptor (KIR), NK inhibitory receptor, NK activating receptor and NK triggering receptor families. We also quantified the frequency of <it>bona fide </it>regulatory T cells (Treg), a T-cell subset implicated in the down-regulation of anti-tumor immunity, and tested the <it>in vitro </it>cytotoxic activity of CIK cells against NK-sensitive, chronic myeloid leukaemia K562 cells.</p> <p>Results</p> <p>CIK cells expanded more vigorously in cultures supplemented with intermediate and high concentrations of TG compared with 50 ng/ml αCD3 mAb. TG-driven CIK cells expressed a constellation of NK activating/inhibitory receptors, such as CD158a and CD158b, NKp46, NKG2D and NKG2A/CD94, released high quantities of IL-12p40 and efficiently lysed K562 target cells. Of interest, the frequency of Treg cells was lower at any time-point compared with PBMC cultures nurtured with αCD3 mAb. Cancer patient-derived CIK cells were also expanded after priming with TG, but they expressed lower levels of the NKp46 triggering receptor and NKG2D activating receptor, thus manifesting a reduced ability to lyse K562 cells.</p> <p>Conclusions</p> <p>TG fosters the generation of functional CIK cells with no concomitant expansion of tumor-suppressive Treg cells. The culture conditions described herein should be applicable to cancer-bearing individuals, although the differentiation of fully functional CIK cells may be hindered in patients with advanced malignancies.</p

Chronic pain and spinal cord stimulation.

Chronic pain can have a devastating impact and lead to patient isolation. Many people with chronic pain are predisposed to anxiety-depressant symptoms, due to a lower quality life. The aim of the study is to demonstrate how neuromodulation methods, can encourage the reduction of chronic pain and an improvement in the quality of life, therefore advancing the restoration of psychological well-being.We involved 50 patients with a diagnosis of pain that not respond to traditional pharmacological therapies. Interventions: All subject had depression and anxiety symptoms and a low-quality life. We used the spinal cord stimulation treatment and a psychological evaluation for assessment of depression-anxiety symptomatology and the level of quality life.We observed a significant difference in physical functioning, role limitations due to physical health, general health perceptions, vitality, social functioning, role limitations due to emotional problems and mental health.Our study affirms that the perception of chronic pain has a great impact on the perception of psychological well-being, quality of life, and the performance of normal daily social and professional activities

Molecular and Technological Characterization of Saccharomyces cerevisiae

The characterization of autochthonous Saccharomyces cerevisiae strains is an important step towards the conservation and employment of microbial biodiversity. The utilization of selected autochthonous yeast strains would be a powerful tool to enhance the organoleptic and sensory properties of typical regional wines. In fact, indigenous yeasts are better tailored to a particular must and because of this they are able to praise the peculiarities of the derived wine. The present study described the biodiversity of indigenous S. cerevisiae strains isolated from natural must fermentations of an ancient and recently rediscovered Apulian grape cultivar, denoted as “Susumaniello.” The yeast strains denoted by the best oenological and technological features were identified and their fermentative performances were tested by either laboratory assay. Five yeast strains showed that they could be excellent candidates for the production of industrial starter cultures, since they dominated the fermentation process and produced wines characterized by peculiar oenological and organoleptic features

Cultural events and tourist services: A management model for guided tours

The introduction of ICT in the tourism sector has had a significant impact on competitiveness that goes way beyond the traditional approach of mere conservation of cultural heritage. Innovative methods in the cultural sector can be useful for tackling the organizational problems and critical issues that affect operators in the various phases of a guided tour. This study analyses the opportunities for the tourism sector and Intangible Cultural Heritage arising from approaches such as Business Process Management (BPM) and Product Lifecycle Management (PLM). These models, already in place in many areas of the industrial sector, could easily be extended to the tourism and cultural sectors. The purpose is to extrapolate a meta-model able to manage the organizational variables and critical issues emerging in the evolution of cultural routes. The model will provide guidelines to the backend unit that organizes guided tours and various kinds of event. This aspect is key to the success of replicable events because it provides a Management Information System that supports future planning with historical data. The management model investigated was applied to a specific case study: “The Ways to Jerusalem: Maritime, Cultural and Pilgrimage Routes, an itinerary of cultural and spiritual interest”

Effects of Alcohol Consumption on Oxidative Stress in a Sample of Patients Recruited in a Dietary Center in a Southern University Hospital: A Retrospective Study

Background and objectives: The aim of this retrospective study was to evaluate the effects of alcohol consumption on oxidative stress. Materials and Methods: The study was conducted by analyzing the increase in lipid peroxidation, the reduction of antioxidant defenses and the alteration of the oxidation/antioxidant balance after the administration of ethanol in 25% aqueous solution (v/v) at a concentration of 0.76 g/kg of body weight daily in two doses for 3 days. The changes in oxidative stress indices were investigated by standard methods previously described. Results: Ethanol administration has determined a significant increase in plasma levels of lipid hydroperoxide (LOOH), malonilaldehyde (MDA) and oxidized glutathione (GSSH), and a decrease in total antioxidant capacity (TAC), reduced glutathione (GSH) and GSH/GSSH ratio. Conclusions: In the proposed experimental condition, the excessive and repeated consumption of ethanol causes oxidative damage, as shown by the increase in lipid peroxidation, the reduction of antioxidant defenses and the alteration of the oxidation/antioxidant balance, which, at least in part, are responsible for the harmful effects of excess ethanol

Effectiveness of high-frequency cervical spinal cord stimulation in the treatment of refractory trigeminal neuropathy: A case report

Rationale: Treatment of chronic neuropathic pain in the head and face regions presents a challenge for pain specialists due to the lack of reliable medical and surgical approaches. Patient concerns: A 62-year-old patient came to our attention for an intense facial pain secondary to a lesion of the right trigeminal nerve (all branches) due to a petroclival meningioma. Diagnoses: The patient also presented with gait impairment as well as a deficit of the right facial, auditory, trochlear and abducens cranial nerves. Interventions: Conventional medical management (CMM) as well as tonic SCS were already adopted but they all dramatically failed. We intervened with the use of high-frequency (10 kHz) spinal cord stimulation (HFSCS) at the cervicomedullary junction (CMJ). The patient was thus provided with HFSCS at the CMJ. Pain and quality of life (QoL) were assessed 1 and 3 months after implantation. We also tested the trigeminal-facial reflex responses. Outcomes: HFSCS led to a full relief from the debilitating electric shocks like pain in the right hemiface, even though a background dull pain appeared. The gradual addition of pregabalin helped in fully relieving the painful symptomatology, with a significant improvement in QoL. Moreover, sensitivity amelioration on the inner portion of the mouth allowed the patient to start feeding again also using that side of the mouth. These findings were paralleled by a significant reshape of trigeminal-facial reflex responses suggesting an inhibition of nociceptive sensory inputs at brainstem level following HFSCS. Lessons: This is the first report suggesting the usefulness of HFSCS at the CMJ in neuropathic pain due to trigeminal nerve neuropathy non-responsive to tonic SCS and CMM