Trigeminal electrical stimulation with ULFTENS of the dorsal anterior mucosal surface of the tongue: Effects on Heart Rate Variability (HRV).

BACKGROUND: Trigeminal electrical stimulation of the dorsal anterior mucosal surface of the tongue has demonstrated its efficacy in a variety of neurological disorders in which anatomical or functional alterations are present. The pathogenesis of such disorders is often linked to altered arousal circuits, and the benefits of tongue stimulation are attributed to the rebalancing of this system. Dental ULFTENS shows efficacy in acting on the muscular, autonomic system and control of the descending pathways that modulate pain. It is administered at the skin level in the area anterior to the tragus and not on the mucosal surface of the tongue. The use of this stimulation technique at the tongue level could have new applications and clinical results if it were able to reduce the activity of arousal circuits. MATERIAL AND METHOD: A new intraoral device allowed electrical stimulation of the dorsal anterior mucosa of the tongue in 32 healthy young women. The effects on HRV were monitored by photoplethysmographic wave (PPG) and compared with a control group. The HRV parameters studied were RMSSD, HF, LF, LF/HF, REC, DET. RESULTS: The group of stimulated subjects showed a significant change in some of the HRV parameters that was maintained even in the epoch after the end of electrical stimulation. This effect can be considered as a vagal activation and a change of HRV trend. The control group of unstimulated subjects showed an opposite trend. There were no undesirable or annoying effects of stimulation. CONCLUSION: Stimulation of the dorsal anterior (trigeminal) mucosal surface of the tongue with ULFTENS applied with an intraoral device was shown to be able to increase HRV.

Electromyographic evaluation of a patient treated with extraoral traction: a case report.

BACKGROUND: This work seeks to provide information on the utility of surface electromyography (SEMG) as an aid for diagnosing orthodontic conditions. Classic orthodontic monitoring by radiography, plaster models, cephalometry, and photography can be improved by using SEMG before and during treatment, to prevent clinical worsening and relapses. CASE REPORT: This paper presents the SEMG results for a 10-year-old female patient, orthodontically treated by extraoral traction (EOT). Significant muscular variations in the patient's EMG were observed as she changed different postures and as headgear device was used. CONCLUSION: SEMG should be performed prior to the orthodontic treatment to assess the neuromuscular patient's pattern, in order to prevent strain induced by extraoral forces. EMG can be a valid aid for evaluating the patient's neuromuscular condition before, during, and after orthodontic treatment.

TENS effects on salivary stress markers: A pilot study.

Transcutaneous electrical nerve stimulation (TENS) is extensively used as pain relief through endorphins release. Moreover, recent findings showed a role in the activation of the autonomic nervous system (ANS); it was evidenced by modification in the heart rate variability and ANS-related marker. The objective of this pilot study is to evaluate salivary alpha amylase (sAA) as a marker of stress in two groups of healthy subjects, one receiving ultra-low frequency transcutaneous electrical nerve stimulation (ULF-TENS) and one without stimulation. Sixty healthy people were enrolled. The test group consisted of 30 participants (15 men, 15 women). The control group consisted of 30 participants (15 men, 15 women). Statistical analysis showed that sAA levels were statistically different between men and women independently from TENS; we hypothesize that treatment could influence sAA levels because it is thought to activate µ opioid receptors. The results of this study seem to indicate that the analysis of sAA, through a non-invasive saliva sample, could be an efficient aid for understanding the functions of the autonomic nervous system.

Investigation of nonlinear pupil dynamics by recurrence quantification analysis.

Pupil is controlled by the autonomous nervous system (ANS). It shows complex movements and changes of size even in conditions of constant stimulation. The possibility of extracting information on ANS by processing data recorded during a short experiment using a low cost system for pupil investigation is studied. Moreover, the significance of nonlinear information contained in the pupillogram is investigated. We examined 13 healthy subjects in different stationary conditions, considering habitual dental occlusion (HDO) as a weak stimulation of the ANS with respect to the maintenance of the rest position (RP) of the jaw. Images of pupil captured by infrared cameras were processed to estimate position and size on each frame. From such time series, we extracted linear indexes (e.g., average size, average displacement, and spectral parameters) and nonlinear information using recurrence quantification analysis (RQA). Data were classified using multilayer perceptrons and support vector machines trained using different sets of input indexes: the best performance in classification was obtained including nonlinear indexes in the input features. These results indicate that RQA nonlinear indexes provide additional information on pupil dynamics with respect to linear descriptors, allowing the discrimination of even a slight stimulation of the ANS. Their use in the investigation of pathology is suggested.

Development of measles virus-based shielded oncolytic vectors: suitability of other paramyxovirus glycoproteins.

Antibody-mediated neutralization may interfere with the efficacy of measles virus (MV) oncolysis. To circumvent vector neutralization, we sought to exchange the envelope glycoproteins, hemagglutinin (H) and fusion (F), with those from the non-crossreactive Tupaia paramyxovirus (TPMV). To sustain efficient particle assembly, we generated hybrid glycoproteins with the MV cytoplasmic tails and the TPMV ectodomains. Hybrid F proteins that partially retained fusion function, and hybrid H proteins that retained fusion support activity, were generated. However, when used in combination, the hybrid proteins did not support membrane fusion. An alternative strategy was developed based on a hybrid F protein and a truncated H protein that supported cell-cell fusion. A hybrid virus expressing these two proteins was rescued, and was able to spread by cell fusion; however, it was only capable of producing minimal amounts of particles. Lack of specific interactions between the matrix and the H protein, in combination with suboptimal F-protein processing and inefficient glycoprotein transport in the rescue cells, accounted for inefficient particle production. Ultimately, this interferes with applications for oncolytic virotherapy. Alternative strategies for the generation of shielded MV are discussed.

Prevalence of myopia in a population with malocclusions.

AIM: The purpose of this study was to investigate the prevalence of myopia among a paediatric population with malocclusions. MATERIALS AND METHODS: A total of 322 consecutive patients of the department of Orthodontics and Gnathology, Dental Clinic, University of L'Aquila, were enlisted for the study and 292 were selected according to the exclusion criteria. Pretreatment diagnostic data, which included radiographic cephalometric and dental cast evaluation, were recorded and presence of myopia was assessed through an ophthalmological examination. Differences in the prevalence of myopia by sex and malocclusion were analysed by using Pearson's chi-square and Fisher's exact tests. RESULTS: According to the sagittal malocclusion, patients were classified as Class I (N=162), Class II division 1 (N=75), Class II division 2 (N=38), or Class III (N=12). No gender influence was found for myopia or malocclusion. No differences were recorded when analysing the influence of sex on the prevalence of myopia in classes of malocclusion. A statistical significant higher prevalence was found for subjects showing myopia in Class II division 1 malocclusion, while no other significant differences were found for prevalence in the other classes of malocclusions. DISCUSSION: Few studies investigated a possible relationship between the ocular and stomatognathic system, and no data are available in the scientific literature. A higher prevalence of myopia was found in patients with Class II division 1: as expected no other significant association was found. CONCLUSION: The findings of the present study suggest a possible association between myopia and Class II, but further studies are needed to confirm and explain this observation.

Armed and targeted measles virus for chemovirotherapy of pancreatic cancer.

No curative therapy is currently available for locally advanced or metastatic pancreatic cancer. Therefore, new therapeutic approaches must be considered. Measles virus (MV) vaccine strains have shown promising oncolytic activity against a variety of tumor entities. For specific therapy of pancreatic cancer, we generated a fully retargeted MV that enters cells exclusively through the prostate stem cell antigen (PSCA). Besides a high-membrane frequency on prostate cancer cells, this antigen is expressed on pancreatic adenocarcinoma, but not on non-neoplastic tissue. PSCA expression levels differ within heterogeneous tumor bulks and between human pancreatic cell lines, and we could show specific infection of pancreatic adenocarcinoma cell lines with both high- and low-level PSCA expression. Furthermore, we generated a fully retargeted and armed MV-PNP-anti-PSCA to express the prodrug convertase purine nucleoside phosphorylase (PNP). PNP, which activates the prodrug fludarabine effectively, enhanced the oncolytic efficacy of the virus on infected and bystander cells. Beneficial therapeutic effects were shown in a pancreatic cancer xenograft model. Moreover, in the treatment of gemcitabine-resistant pancreatic adenocarcinoma cells, no cross-resistance to both MV oncolysis and activated prodrug was detected.

CD20-targeted measles virus shows high oncolytic specificity in clinical samples from lymphoma patients independent of prior rituximab therapy.

New therapeutic modalities for B-cell non-Hodgkin's lymphomas (B-NHL) are needed, especially for relapsing and aggressive subtypes. Toward this end, we previously generated a fully CD20-targeted and armed measles virus, and tested its efficacy in a xenograft model of mantle cell lymphoma (MCL). Here, we quantify its spread in peripheral blood mononuclear cells and/or tissue of patients with different histological subtypes of B-NHL, including splenic marginal zone lymphoma (SMZL). CD20-targeted MV efficiently infects lymphoma cells from SMZL and MCL while sparing most cells in the CD20-negative population, in contrast to the parental vaccine-lineage MV, which infects CD20-positive and CD20-negative cells equally. Rituximab therapy (4-8 months before relapse) did not interfere with the infectivity and specificity of MV(green)H(blind)antiCD20 in patient lymphoma samples. Thus, CD20-targeted oncolytic virotherapy is likely to be effective after previous antiCD20 therapy.

Mantle cell lymphoma salvage regimen: synergy between a reprogrammed oncolytic virus and two chemotherapeutics.

Measles virus (MV)-PNP H(blind)antiCD20 is a CD20-targeted and prodrug convertase-armed MV that temporarily controls growth of lymphoma xenografts in severe combined immunodeficiency (SCID) mice in combination with fludarabine phosphate (fludarabine). Herein, we examine the replication of this targeted virus and of a vaccine-lineage MV in disease bulks and circulating cells from mantle cell lymphoma (MCL) patients, and show that only the targeted virus is specific for CD20-expressing cells. We then assessed the efficacy of different regimens of administration of this virus in combination with fludarabine and cyclophosphamide (CPA) in an MCL xenograft model. We show that CPA administration before the beginning of virus treatment enhances oncolytic efficacy, likely through temporary immunosuppression. An interval of 1 week between intravenous virus administration and fludarabine treatment further enhanced oncolysis, by synchronizing maximum prodrug convertase expression with fludarabine availability. Finally, three 23-day courses of triple sequential treatment with CPA, virus and fludarabine treatment resulted in complete regression of the xenografts. Secondary disease symptoms interfered with survival, but average survival times increased from 22 to 77 days. These studies document a reprogrammed oncolytic virus, consolidating the effects of two chemotherapeutics, a concept well suited for a phase I clinical trial for MCL patients for whom conventional therapies have failed.

Effects of myogenous facial pain on muscle activity of head and neck.

This study evaluated the effects of visual input on surface electromyography (sEMG) of some stomathognatic and neck muscles (anterior temporalis, masseter, anterior digastric and sternocleidomastoid muscles) in patients experiencing myogenous facial pain compared with healthy volunteers. All subjects kept the mandible at rest with teeth apart and underwent a 15-s sEMG recording of anterior temporalis, masseter, digastric and sternocleidomastoid muscles. Each recording was carried out with closed and then open eyes. The sEMG activity of each muscle was compared between the two groups. In the study group, anterior temporalis, masseter and sternocleidomastoid sEMG with closed eyes showed higher values compared with controls (p<0.05). In the study group, left and right anterior temporalis (p<0.003) and right digastric (p<0.03) sEMG with open eyes showed higher values than sEMG with closed eyes. In the control group no significant differences were observed between closed and open eyes. In patients with myogenous facial pain, visual input appears to be associated with a significant increase in the sEMG activity of some head and neck muscles.

[Antiphospholipid antibody: laboratory, pathogenesis and clinical manifestations]. / Gli anticorpi antifosfolipidi: il laboratorio, la patogenesi e la clinica (cosa è utile sapere degli anticorpi antifosfolipidi).

Antiphospholipid antibodies (aPL) represent a heterogeneous group of antibodies that recognize various antigenic targets including beta2 glycoprotein I (beta2GPI), prothrombin (PT), activated protein C, tissue plasminogen activator, plasmin and annexin A2. The most commonly used tests to detect aPL are: lupus anticoagulant (LAC), a functional coagulation assay, anticardiolipin antibody (aCL) and anti-beta2GPI antibody (anti-beta2GPI), which are enzyme-linked immunoassay (ELISA). Clinically aPL are associated with thrombosis and/or with pregnancy morbidity. Apparently aPL alone are unable to induce thrombotic manifestations, but they increase the risk of vascular events that can occur in the presence of another thrombophilic condition; on the other hand obstetrical manifestations were shown to be associated not only to thrombosis but mainly to a direct antibody effect on the trophoblast.

Antiphospholipid antibody profile: implications for the evaluation and management of patients.

According to the classification criteria of antiphospholipid syndrome, lupus anticoagulant, anticardiolipin and anti-beta(2) glycoprotein I antibody assays are independent risk factors for the occurrence of vascular thrombosis and pregnancy loss. However, it is generally accepted that patients carrying multiple positivity have more a severe disease and higher recurrence rate despite treatment. On the other hand, the diagnostic value of a positive result in one only assay is more controversial, particularly in the presence of clinical manifestations such as deep vein thrombosis or early miscarriages, which are rather common in the general population. In this review we speculate on current and future strategies to interpret different antiphospholipid antibody profiles in the clinical practice.

Antiphospholipid antibodies and malignancies.

Since the 1980s it is known that an important thrombogenic mechanism is mediated by antiphospholipid antibodies (aPL). Aim of this review is to discuss how much aPL presence may worsen the thrombophilic state of neoplastic patients and how much cancer may worsen and extend the thrombophilic state of patients with Antiphospholipid Syndrome (APS). In the last years a higher prevalence of aPL was observed in patients with solid tumors compared to controls. These patients, already at higher risk of thrombosis, may have a still higher risk when aPL carriers. Those with a solid malignancy seem to be more likely to have a thrombotic event compared to patients with a hematological disorder. On the other hand aPL presence may be a risk factor for malignancies (particularly hematological). Even if the significance of aPL and cancer relationship has to be further investigated, clinicians should remember that in neoplastic patients aPL presence can increase thromboembolic risk and in healthy carriers can increase the possibility of developing a malignancy.

The 10-year follow-up data of the Euro-Lupus Nephritis Trial comparing low-dose and high-dose intravenous cyclophosphamide.

OBJECTIVE: To update the follow-up of the Euro-Lupus Nephritis Trial (ELNT), a randomised prospective trial comparing low-dose (LD) and high-dose (HD) intravenous (IV) cyclophosphamide (CY) followed by azathioprine (AZA) as treatment for proliferative lupus nephritis. PATIENTS AND METHODS: Data for survival and kidney function were prospectively collected during a 10-year period for the 90 patients randomised in the ELNT, except in 6 lost to follow-up. RESULTS: Death, sustained doubling of serum creatinine and end-stage renal disease rates did not differ between the LD and HD group (5/44 (11%) vs 2/46 (4%), 6/44 (14%) vs 5/46 (11%) and 2/44 (5%) vs 4/46 (9%), respectively) nor did mean serum creatinine, 24 h proteinuria and damage score at last follow-up. Most patients in both groups were still treated with glucocorticoids, other immunosuppressant agents and blood pressure lowering drugs. After 10 years of follow-up, the positive predictive value for a good outcome of an early drop in proteinuria in response to initial immunosuppressive therapy was confirmed. CONCLUSION: The data confirm that a LD IVCY regimen followed by AZA-the "Euro-Lupus regimen"-achieves good clinical results in the very long term.

Effect on anterior temporalis surface EMG of eyes open-closed condition.

BACKGROUND: The use of rest surface EMG of jaw elevator muscles is still debated. The low voltage recorded in anterior temporalis muscle by electromyography (EMG) in rest position could be affected by electronic noise or by activity coming from other muscles. Our goal was to evaluate the physiological behaviour of the anterior temporalis by surface EMG at rest mandible position during open or closed eyes condition in healthy young subjects without both malocclusion and visual defect. MATERIALS AND METHODS: Surface EMG of anterior temporalis, masseter, digastric, sternomastoid muscle and mandible kinesiographic movement were recorded in 20 young, healthy individuals without both malocclusion and visual defect during open-closed eyes condition. RESULTS: No significant difference was found in surface EMG of anterior temporalis comparing eyes closed to eyes open condition. CONCLUSION: Physiology of open-closed eyes in healthy, young subjects without malocclusion or visual defect does not imply a change in surface EMG of anterior temporalis muscle.

[Methylenetetrahydrofolate reductase polymorphisms in methotrexate treatment of rheumatoid arthritis patients. Review of the literature and personal experience]. / I polimorfismi della metilentetraidrofolatoreduttasi (MTHFR) nel trattamento con methotrexate di pazienti con artrite reumatoide. Revisione della letteratura ed esperienza personale.

Methotrexate is still a mainstay of rheumatoid arthritis treatment, but a significant variability in drug response is observed among patients. It has been proposed that C677T and A1298C polymorphisms of methylenetetrahydrofolate reductase (MTHFR), an enzyme involved in the folate pathway, could be related to its efficacy and toxicity. Many studies have investigated the predictive value of such polymorphisms for Methotrexate outcome, though with discordant results. Our experience on 79 patients did not find any significant association between genotype and drug response and the review of the literature did not provide sufficient evidences to support the use of MTHFR genetic screening in clinical practice.

Methylenetetrahydrofolate reductase polymorphisms and methotrexate: no association with response to therapy nor with drug-related adverse events in an Italian population of rheumatic patients.

OBJECTIVE: MTHFR is an enzyme involved in the folate pathway. It has been suggested that common polymorphisms in its gene (C677T and A1298C) could be related to different methotrexate (MTX) response and toxicity in rheumatoid arthritis (RA) patients. Agreement has not been found yet and there is no data on rheumatic Italian patients. The aim of this study is to determine if a genetic screening can help in planning the treatment in these patients. METHODS: We enrolled 84 Northern Italian patients affected by RA (n=79), psoriatic arthritis (n=4) and ankylosing spondylitis (n=1), who received MTX. Subjects who achieved at least ACR20 response in 6 months and maintained it during the following 6 months were defined as "responders"; those who did not obtain a disease control after 6 months of MTX were classified as "non responders". Patients who experienced MTX adverse events were defined "with toxicity", those who did not, as "without toxicity". Genotypes were determined by polymerase chain reaction. RESULTS: Genotype frequency was consistent with that reported in a healthy population from Italy. We did not find any statistically significant difference in genotype/allele distribution between the groups. In patients receiving folic acid supplementation MTX toxicity was recorded only in 18 cases (24%), while all the 8 patients not receiving it experienced MTX adverse events (p=0.00). CONCLUSION: In our study we did not find any association between MTHFR genotype/allele and MTX response or toxicity. At the moment there is not sufficient evidence for MTHFR screening in patients who are candidate for MTX.

Treatment of lupus skin involvement with quinacrine and hydroxychloroquine.

To evaluate the efficacy of hydroxychloroquine (HCQ) and quinacrine (Qn) association, at two different dosages, in treatment of lupus skin lesions not responding to HCQ alone. Thirty-four patients, affected by cutaneous and systemic lupus erythematosus, were retrospectively analysed. They were treated by HCQ (5 mg/Kg/qd) and Qn with two regimens: 100 mg/qd (29 cases) and 50 mg/qd (5 cases). Discoid lupus erythematosus (19 cases), acute malar rash (6 cases), chilblain lupus (4 cases) showed a significant improvement with combination therapy (P = 0.009, P = 0.019, and P = 0.04, respectively). Ten patients with subacute cutaneous lupus showed a partial response, whereas lupus profundus didn't improve. The same overall response rate was recorded comparing two Qn regimens, but subjects taking 100 mg/qd improved more rapidly than the others (P = 0.001). Ten patients developed side effects, mainly represented by skin yellowish discolouration. Depression and severe headache with nausea, which were globally recorded in two cases, led to drug withdrawal. One additional case of hepatitis was recorded in a patient with preexisting Hepatitis C virus (HCV) infection. Combination of HCQ and Qn is rapidly effective at 100 mg/qd and well tolerated in the treatment of lupus skin lesions unresponsive to HCQ alone.

Pseudotyping lentiviral vectors with the wild-type measles virus glycoproteins improves titer and selectivity.

We pseudotyped HIV-1 vectors with cytoplasmic tail-truncated envelope glycoproteins of a wild-type (WT) measles virus (MV). The particles entered the lymphatic cells exclusively through the signaling lymphocyte activation molecule (SLAM, CD150), whereas particles pseudotyped with the MV vaccine strain glycoproteins also recognized the ubiquitous membrane cofactor protein (CD46) as receptor and had less specific cell entry. MV(WT)-HIV vectors reached titers of 10(8) t.u. ml(-1), which were up to 10-fold higher than those of MV(Vac)-HIV vectors, and discriminated between SLAM-positive and SLAM-negative cells, also in mixed cell cultures. As these vectors transduce primary human cells more efficiently than vesicular stomatitis virus-G pseudotyped vectors do, they are promising candidates for gene transfer to human lymphocytes and certain epithelial cells.

Measles virus glycoprotein complex assembly, receptor attachment, and cell entry.

Measles virus (MV) enters cells by membrane fusion at the cell surface at neutral pH. Two glycoproteins mediate this process: the hemagglutinin (H) and fusion (F) proteins. The H-protein binds to receptors, while the F-protein mediates fusion of the viral and cellular membranes. H naturally interacts with at least three different receptors. The wild-type virus primarily uses the signaling lymphocyte activation molecule (SLAM, CD150) expressed on certain lymphatic cells, while the vaccine strain has gained the ability to also use the ubiquitous membrane cofactor protein (MCP, CD46), a regulator of complement activation. Additionally, MV infects polarized epithelial cells through an unidentified receptor (EpR). The footprints of the three receptors on H have been characterized, and the focus of research is shifting to the characterization of receptor-specific conformational changes that occur in the H-protein dimer and how these are transmitted to the F-protein trimer. It was also shown that MV attachment and cell entry can be readily targeted to designated receptors by adding specificity determinants to the H-protein. These studies have contributed to our understanding of membrane fusion by the glycoprotein complex of paramyxoviruses in general.