Dalfampridine in patients with downbeat nystagmus—an observational study

We investigated the effects of dalfampridine, the sustained-release form of 4-aminopyridine, on slow phase velocity (SPV) and visual acuity (VA) in patients with downbeat nystagmus (DBN) and the side effects of the drug. In this proof-of-principle observational study, ten patients received dalfampridine 10 mg bid for 2 weeks. Recordings were conducted at baseline, 180 min after first administration, after 2 weeks of treatment and after 4 weeks of wash-out. Mean SPV decreased from a baseline of 2.12 deg/s ± 1.72 (mean ± SD) to 0.51 deg/s ± 1.00 180 min after first administration of dalfampridine 10 mg and to 0.89 deg/s ± 0.75 after 2 weeks of treatment with dalfampridine (p < 0.05; post hoc both: p < 0.05). After a wash-out period of 1 week, mean SPV increased to 2.30 deg/s ± 1.6 (p < 0.05; post hoc both: p < 0.05). The VA significantly improved during treatment with dalfampridine. Also, 50 % of patients did not report any side effects. The most common reported side effects were abdominal discomfort and dizziness. Dalfampridine is an effective treatment for DBN in terms of SPV. It was well-tolerated in all patients.

Tritrichomonas foetus: Prevalence study in naturally mating bulls in Switzerland

Switzerland is officially free from bovine Tritrichomonas foetus. While bulls used for artificial insemination (AI) are routinely examined for this pathogen, bulls engaged in natural mating, as well as aborted fetuses, are only very sporadically investigated, indicating that the disease awareness for bovine tritrichomoniasis is low. Natural mating in cattle is becoming increasingly popular in Switzerland. Accordingly, a re-introduction/re-occurrence of T. foetus in cattle seems possible either via resurgence from a yet unknown bovine reservoir, or via importation of infected cattle. The low disease awareness for bovine tritrichomoniasis might favor an unnoticed re-establishment of T. foetus in the Swiss cattle population. The aim of our study was thus to search for the parasite, and if found, to assess the prevalence of bovine T. foetus in Switzerland. We included (1) bulls over two years of age used in natural mating and sent to slaughter, (2) bulls used for natural service in herds with or without fertility problems and (3) aborted fetuses. Furthermore, the routinely examined bulls used for AI (4) were included in this study. In total, 1362 preputial samples from bulls and 60 abomasal fluid samples of aborted fetuses were analyzed for the presence of T. foetus by both in vitro cultivation and molecular analyses. The parasite could not be detected in any of the samples, indicating that the maximal prevalence possibly missed was about 0.3% (95% confidence). Interestingly, in preputial samples of three bulls of category 1, apathogenic Tetratrichomonas sp. was identified, documenting a proof-of-principle for the methodology used in this study.

The Glucose-Dependent Insulinotropic Polypeptide Receptor: A Novel Target for Neuroendocrine Tumor Imaging-First Preclinical Studies

A new family of peptide receptors, the incretin receptor family, overexpressed on many neuroendocrine tumors (NETs) is of great importance because it may enable the in vivo peptide-based receptor targeting of a category of NETs that does not express the somatostatin receptor. Impressive in vivo diagnostic data were published for glucagonlike peptide 1 receptor-targeting radiopeptides. Recently, promising in vitro data have appeared for the second member of the incretin family, the glucose-dependent insulinotropic polypeptide (GIP) receptor. This prompted us to develop and evaluate a new class of radioligands with the potential to be used for the in vivo targeting of GIP receptor-positive tumors. METHODS GIP(1-42) was modified C-terminally, and the truncated peptides [Lys(30)(aminohexanoic acid [Ahx]-DOTA)]GIP(1-30)NH2 (EG1), [Lys(16)(Ahx-DOTA)]GIP(1-30)NH2 (EG2), and [Nle(14), Lys(30)(Ahx-DOTA)]GIP(1-30)NH2 (EG4) were conjugated with Ahx-DOTA via the Lys(16) and Lys(30) side chains. Their inhibitory concentration of 50% (IC50) was determined using [(125)I-Tyr(10)]GIP(1-30) as radioligand and GIP(1-30) as control peptide. The DOTA conjugates were labeled with (111)In and (68)Ga. In vitro evaluation included saturation and internalization studies using the pancreatic endocrine cell line INR1G9 transfected with the human GIP receptor (INR1G9-hGIPr). The in vivo evaluation consisted of biodistribution and PET imaging studies on nude mice bearing INR1G9-hGIPr tumors. RESULTS Binding studies (IC50 and saturation studies) showed high affinity toward GIP receptor for the GIP conjugates. Specific in vitro internalization was found, and almost the entire cell-associated activity was internalized (>90% of the cell-bound activity), supporting the agonist potency of the (111)In-vectors. (111)In-EG4 and (68)Ga-EG4 were shown to specifically target INR1G9-hGIPr xenografts, with tumor uptake of 10.4% ± 2.2% and 17.0% ± 4.4% injected activity/g, 1 h after injection, respectively. Kidneys showed the highest uptake, which could be reduced by approximately 40%-50% with a modified-fluid-gelatin plasma substitute or an inhibitor of the serine protease dipeptidyl peptidase 4. The PET images clearly visualized the tumor. CONCLUSION The evaluation of EG4 as a proof-of-principle radioligand indicated the feasibility of imaging GIP receptor-positive tumors. These results prompt us to continue the development of this family of radioligands for imaging of a broad spectrum of NETs.

GABAergic modulation in central sensitization in humans: a randomized placebo-controlled pharmacokinetic-pharmacodynamic study comparing clobazam with clonazepam in healthy volunteers

Positive allosteric modulators of GABAA receptors (GAMs) acting at specific subtypes of GABAA receptors effectively restore compromised spinal pain control in rodents. Studies addressing a similar antihyperalgesic effect in humans are sparse and are hampered by sedative effects of nonselective GAMs available for use in humans. We present results from a randomized controlled double-blind crossover study in 25 healthy volunteers, which addressed potential antihyperalgesic actions of clobazam (CBZ) and clonazepam (CLN) at mildly sedating equianticonvulsive doses. Clobazam was chosen because of its relatively low sedative properties and CLN because of its use in neuropathic pain. Tolterodine (TLT) was used as an active placebo. The primary outcome parameter was a change in the area of cutaneous UVB irradiation-induced secondary hyperalgesia (ASH), which was monitored for 8 hours after drug application. Sedative effects were assessed in parallel to antihyperalgesia. Compared with TLT, recovery from hyperalgesia was significantly faster in the CBZ and CLN groups (P = 0.009). At the time point of maximum effect, the rate of recovery from hyperalgesia was accelerated by CBZ and CLN, relative to placebo by 15.7% (95% confidence interval [CI] 0.8-30.5), P = 0.040, and 28.6% (95% CI 4.5-52.6), P = 0.022, respectively. Active compounds induced stronger sedation than placebo, but these differences disappeared 8 hours after drug application. We demonstrate here that GAMs effectively reduce central sensitization in healthy volunteers. These results provide proof-of-principle evidence supporting efficacy of GAMs as antihyperalgesic agents in humans and should stimulate further research on compounds with improved subtype specificity.

The virtual patient in dental medicine

OBJECTIVES The aim of this Short Communication was to present a workflow for the superimposition of intraoral scan (IOS), cone-beam computed tomography (CBCT), and extraoral face scan (EOS) creating a 3D virtual dental patient. MATERIAL AND METHODS As a proof-of-principle, full arch IOS, preoperative CBCT, and mimic EOS were taken and superimposed to a unique 3D data pool. The connecting link between the different files was to detect existing teeth as constant landmarks in all three data sets. RESULTS This novel application technique successfully demonstrated the feasibility of building a craniofacial virtual model by image fusion of IOS, CBCT, and EOS under 3D static conditions. CONCLUSIONS The presented application is the first approach that realized the fusion of intraoral and facial surfaces combined with skeletal anatomy imaging. This novel 3D superimposition technique allowed the simulation of treatment planning, the exploration of the patients' expectations, and the implementation as an effective communication tool. The next step will be the development of a real-time 4D virtual patient in motion.

Dielectrophoresis-based analyte separation and analysis

Dielectrophoresis—the tendency of a material of high dielectric permittivity to migrate in an electrical field gradient to a region of maximum field strength—provides an ideal motive force for manipulating small volumes of biological analytes in microfluidic microsystems. The work described in this thesis was based on the hypothesis that dielectrophoresis could be exploited to provide high-resolution cell separations in microsystems as well as a means for the electrically-controllable manipulation of solid supports for molecular analysis. To this end, a dielectrophoretic/gravitational field-flow-fractionation (DEP/G-FFF) system was developed and the separation performance evaluated using various types and sizes of polystyrene microspheres as model particles. It was shown that separation of the polystyrene beads was based on the differences in their effective dielectrophoretic properties. The ability of an improved DEP/G-FFF system to separate genetically identical, but phenotypically dissimilar cell types was demonstrated using mixtures of 6m2 mutant rat kidney cells grown under transforming and non-transforming culture conditions. Additionally, a panel of engineered dielectric microspheres was designed with specific, predetermined dielectrophoretic properties such that their dielectrophoretic behaviors would be controllable and predictable. The fabrication method involved the use of gold-coated polystyrene microsphere cores coated with a self-assembled monolayer of alkanethiol and, optionally, a self-assembled monolayer of phospholipid to form a thin-insulating-shell-over-conductive-interior structure. The successful development of the DEP/G-FFF separation system and the dielectrically engineered microspheres provides proof-of-principle demonstrations of enabling dielectrophoresis-based microsystem technology that should provide powerful new methods for the manipulation, separation and identification of analytes in many diverse fields. ^

Antigen-specific proteolytic antibodies

The antigen recognition site of antibodies is composed of residues contributed by the variable domains of the heavy and light chain subunits (VL and VH domains). VL domains can catalyze peptide bond hydrolysis independent of VH domains (Mei S et al. J Biol Chem. 1991 Aug 25;266(24):15571-4). VH domains can bind antigens noncovalently independent of V L domains (Ward et al. Nature. 1989 Oct 12;341(6242):544-6). This dissertation describe the specific hydrolysis of fusion proteins containing the hepatitis C virus coat protein E2 by recombinant hybrid Abs composed of the heavy chain of a high affinity anti-E2 IgG1 paired with light chains expressing promiscuous catalytic activity. The proteolytic activity was evident from electrophoresis assays using recombinant E2 substrates containing glutathione S-transferase (E2-GST) or FLAG peptide (E2-FLAG) tags. The proteolytic reaction proceeded more rapidly in the presence of the hybrid IgG1 compared to the unpaired light chain, consistent with accelerated peptide bond hydrolysis due to noncovalent VH domain-E2 recognition. An active site-directed inhibitor of serine proteases inhibited the proteolytic activity of the hybrid IgG, indicating a serine protease mechanism. Binding studies confirmed that the hybrid IgG retained detectable noncovalent E2 recognition capability, although at a level smaller than the wildtype anti-E2 IgG. Immunoblotting of E2-FLAG treated with the hybrid IgG suggested a scissile bond within E2 located ∼11 kD from the N terminus of the protein. E2-GST was hydrolyzed by the hybrid IgG at peptide bonds located in the GST tag. The differing cleavage pattern of E2-FLAG and E2-GST can be explained by the split-site model of catalysis, in which conformational differences in the E2 fusion protein substrates position alternate peptide bonds in register with the antibody catalytic subsite despite a common noncovalent binding mechanism. This is the first proof-of principle that the catalytic activity of a light chain can be rendered antigen-specific by pairing with a noncovalently binding heavy chain subunit. ^

He isotopes in Late Cretaceous sediments

Late Cretaceous (100-73 Ma) pelagic limestones were measured for helium concentration and isotopic composition to characterize the interplanetary dust flux using 3He as a tracer. In the Bottaccione section near Gubbio, Italy, three intervals of elevated 3He concentration were detected: K1 in the Campanian stage at ~79 Ma, K2 in the Santonian stage at ~ 85 Ma, and K3 in the Turonian stage at ~91 Ma. All three of these episodes are associated with high 3He/4He and 3He/non-carbonate ratios, consistent with their derivation from an enhanced extraterrestrial 3He flux rather than decreased carbonate sedimentation or dissolution. While K2 is modest in magnitude and duration and thus is of limited significance, K1 and K3 are each identified by a few myr interval with an ~4-fold enhancement in mean 3He flux compared with pre-event levels. Samples from ODP Hole 762C in the Indian Ocean spanning both K2 and K3 (93-83 Ma) confirm the presence of a peak in the Turonian stage, suggesting that K3 is a global event. The K1 and K3 3He events are similar in most respects to the two peaks previously detected in the Cenozoic, suggesting a similar origin. These have been attributed to a major asteroid collision in the Late Miocene and to a shower of either comets or asteroids in the Late Eocene. Based on the age and temporal evolution of K1, we suggest that it most likely records the collision which produced the Baptistina asteroid family independently dated at ~80 Ma. The K3 event is less easily explained. It is characterized by an unusually spiky and erratic temporal progression, suggesting an unusual abundance of very 3He rich particles not previously seen in the sedimentary 3He record. We suggest this episode arises either from a comet shower or from an asteroid shower possibly associated with dust-producing lunar impacts.

Steps Ahead in the Few-Group CRoss-Section Library Generation at the Pin Level

There exists an interest in performing pin-by-pin calculations coupled with thermal hydraulics so as to improve the accuracy of nuclear reactor analysis. In the framework of the EU NURISP project, INRNE and UPM have generated an experimental version of a few group diffusion cross sections library with discontinuity factors intended for VVER analysis at the pin level with the COBAYA3 code. The transport code APOLLO2 was used to perform the branching calculations. As a first proof of principle the library was created for fresh fuel and covers almost the full parameter space of steady state and transient conditions. The main objective is to test the calculation schemes and post-processing procedures, including multi-pin branching calculations. Two library options are being studied: one based on linear table interpolation and another one using a functional fitting of the cross sections. The libraries generated with APOLLO2 have been tested with the pin-by-pin diffusion model in COBAYA3 including discontinuity factors; first comparing 2D results against the APOLLO2 reference solutions and afterwards using the libraries to compute a 3D assembly problem coupled with a simplified thermal-hydraulic model.

Silicon Photomultipliers (SiPM) as novel photodetectors for PET

Next generation PET scanners should fulfill very high requirements in terms of spatial, energy and timing resolution. Modern scanner performances are inherently limited by the use of standard photomultiplier tubes. The use of Silicon Photomultipliers (SiPMs) is proposed for the construction of a 4D-PET module of 4.8×4.8 cm2 aimed to replace the standard PMT based PET block detector. The module will be based on a LYSO continuous crystal read on two faces by Silicon Photomultipliers. A high granularity detection surface made by SiPM matrices of 1.5 mm pitch will be used for the x–y photon hit position determination with submillimetric accuracy, while a low granularity surface constituted by 16 mm2 SiPM pixels will provide the fast timing information (t) that will be used to implement the Time of Flight technique (TOF). The spatial information collected by the two detector layers will be combined in order to measure the Depth of Interaction (DOI) of each event (z). The use of large area multi-pixel Silicon Photomultiplier (SiPM) detectors requires the development of a multichannel Data Acquisition system (DAQ) as well as of a dedicated front-end in order not to degrade the intrinsic detector capabilities and to manage many channels. The paper describes the progress made on the development of the proof of principle module under construction at the University of Pisa.

Wearable soft robotic device for post-stroke shoulder rehabilitation: identifying misalignments

Stroke is the leading cause of long-term disability in the United States, affecting over 795,000 people annually. In order to regain motor function of the upper body, patients are usually treated by regular sessions with a dedicated physical therapist. A cost-effective wearable upper body orthotics system that can be used at home to empower both the patients and physical therapists is described. The system is composed of a thin, compliant, lightweight, cost-effective soft orthotic device with an integrated cable actuation system that is worn over the upper body, an embedded limb position sensing system, an electric actuator package and controller. The proposed device is robust to misalignments that may occur during actuation of the compliant brace or when putting on the system. Through simulations and experimental evaluation, it was demonstrated i) that the soft orthotic cable-driven shoulder brace can be successfully actuated without the production of off-axis torques in the presence of misalignments and ii) that the proposed model can identify linear and angular misalignments online.

PÓS-GRADUAÇÃO, PRODUÇÃO ACADÊMICA, INTERDISCIPLINARIDADE E SOCIALIZAÇÃO DO CONHECIMENTO: A PRODUÇÃO CIENTÍFICA DO GRANDE ABC À LUZ DO PNPG 2011-2020

O objetivo desta investigação foi verificar se, diante da autonomia e vocação que possuem, bem como da possibilidade de receber amparo e incentivos governamentais, as universidades pertencentes à região do Grande ABC atenderam as recomendações feitas pelo PNPG vigente. Para tanto, foram analisadas cinquenta e sete dissertações e duas teses da área de ciências sociais aplicadas, publicadas no período entre 2011 e 2014, pelas seguintes instituições: Universidade Metodista de São Paulo (UMESP); Universidade Municipal de São Caetano do Sul (USCS); Universidade Federal do ABC (UFABC) e Faculdade de Engenharia Industrial (FEI). Essa averiguação se deu ao redor de dois eixos organizadores do PNPG 2011-2014: o terceiro eixo – o aperfeiçoamento da avaliação e sua expansão para outros segmentos do sistema de CT&I (formação de pós-graduados voltados para atividades extra-acadêmicas/setor empresarial) e o quarto eixo – a multi e a interdisciplinaridade entre as principais características da pós-graduação e importantes temas da pesquisa (promover, por meio de programas, áreas de concentração e linhas de pesquisa, a convergência de temas e compartilhamento de problemas em oposição à sua mera associação ou sobreposição). Este estudo – qualitativo, bibliográfico, documental, exploratório, descritivo, tipo estado do conhecimento – se desenvolveu por meio de pesquisa documental e de análise de conteúdo temático categorial. A coleta de dados foi feita por meio dos repositórios digitais de teses e dissertações mantidos na internet pelas Universidades, cuja produção científica foi investigada. Após a análise dos dados ficou demonstrado que aproximadamente 68,96% da produção científica da área de ciências sociais aplicadas, publicada pelas universidades do Grande ABC, no período entre 2011 e 2014, corresponde às expectativas do PNPG atual no que diz respeito às recomendações constantes no terceiro eixo. Em relação às recomendações feitas no texto do quarto eixo, vemos que aproximadamente 31,03% dos trabalhos selecionados atendem às expectativas do Plano, apresentando em sua estrutura, segundo a fundamentação teórica utilizada neste trabalho, características de interdisciplinaridade.

PÓS-GRADUAÇÃO, PRODUÇÃO ACADÊMICA, INTERDISCIPLINARIDADE E SOCIALIZAÇÃO DO CONHECIMENTO: A PRODUÇÃO CIENTÍFICA DO GRANDE ABC À LUZ DO PNPG 2011-2020

O objetivo desta investigação foi verificar se, diante da autonomia e vocação que possuem, bem como da possibilidade de receber amparo e incentivos governamentais, as universidades pertencentes à região do Grande ABC atenderam as recomendações feitas pelo PNPG vigente. Para tanto, foram analisadas cinquenta e sete dissertações e duas teses da área de ciências sociais aplicadas, publicadas no período entre 2011 e 2014, pelas seguintes instituições: Universidade Metodista de São Paulo (UMESP); Universidade Municipal de São Caetano do Sul (USCS); Universidade Federal do ABC (UFABC) e Faculdade de Engenharia Industrial (FEI). Essa averiguação se deu ao redor de dois eixos organizadores do PNPG 2011-2014: o terceiro eixo – o aperfeiçoamento da avaliação e sua expansão para outros segmentos do sistema de CT&I (formação de pós-graduados voltados para atividades extra-acadêmicas/setor empresarial) e o quarto eixo – a multi e a interdisciplinaridade entre as principais características da pós-graduação e importantes temas da pesquisa (promover, por meio de programas, áreas de concentração e linhas de pesquisa, a convergência de temas e compartilhamento de problemas em oposição à sua mera associação ou sobreposição). Este estudo – qualitativo, bibliográfico, documental, exploratório, descritivo, tipo estado do conhecimento – se desenvolveu por meio de pesquisa documental e de análise de conteúdo temático categorial. A coleta de dados foi feita por meio dos repositórios digitais de teses e dissertações mantidos na internet pelas Universidades, cuja produção científica foi investigada. Após a análise dos dados ficou demonstrado que aproximadamente 68,96% da produção científica da área de ciências sociais aplicadas, publicada pelas universidades do Grande ABC, no período entre 2011 e 2014, corresponde às expectativas do PNPG atual no que diz respeito às recomendações constantes no terceiro eixo. Em relação às recomendações feitas no texto do quarto eixo, vemos que aproximadamente 31,03% dos trabalhos selecionados atendem às expectativas do Plano, apresentando em sua estrutura, segundo a fundamentação teórica utilizada neste trabalho, características de interdisciplinaridade.

Compact stellarators with modular coils

Compact stellarator designs with modular coils and only two or three field periods are now available; these designs have both good stability and quasiaxial symmetry providing adequate transport for a magnetic fusion reactor. If the bootstrap current assumes theoretically predicted values a three field period configuration is optimal, but if that net current turns out to be lower, a device with two periods and just 12 modular coils might be better. There are also attractive designs with quasihelical symmetry and four or five periods whose properties depend less on the bootstrap current. Good performance requires that there be a satisfactory magnetic well in the vacuum field, which is a property lacking in a stellarator-tokamak hybrid that has been proposed for a proof of principle experiment. In this paper, we present an analysis of stability for these configurations that is based on a mountain pass theorem asserting that, if two solutions of the problem of magnetohydrodynamic equilibrium can be found, then there has to be an unstable solution. We compare results of our theory of equilibrium, stability, and transport with recently announced measurements from the large LHD experiment in Japan.

In vivo activity in a catalytic antibody-prodrug system: Antibody catalyzed etoposide prodrug activation for selective chemotherapy

Effective chemotherapy remains a key issue for successful cancer treatment in general and neuroblastoma in particular. Here we report a chemotherapeutic strategy based on catalytic antibody-mediated prodrug activation. To study this approach in an animal model of neuroblastoma, we have synthesized prodrugs of etoposide, a drug widely used to treat this cancer in humans. The prodrug incorporates a trigger portion designed to be released by sequential retro-aldol/retro-Michael reactions catalyzed by aldolase antibody 38C2. This unique prodrug was greater than 102-fold less toxic than etoposide itself in in vitro assays against the NXS2 neuroblastoma cell line. Drug activity was restored after activation by antibody 38C2. Proof of principle for local antibody-catalyzed prodrug activation in vivo was established in a syngeneic model of murine neuroblastoma. Mice with established 100-mm3 s.c. tumors who received one intratumoral injection of antibody 38C2 followed by systemic i.p. injections with the etoposide prodrug showed a 75% reduction in s.c. tumor growth. In contrast, injection of either antibody or prodrug alone had no antitumor effect. Systemic injections of etoposide at the maximum tolerated dose were significantly less effective than the intratumoral antibody 38C2 and systemic etoposide prodrug combination. Significantly, mice treated with the prodrug at 30-fold the maximum tolerated dose of etoposide showed no signs of prodrug toxicity, indicating that the prodrug is not activated by endogenous enzymes. These results suggest that this strategy may provide a new and potentially nonimmunogenic approach for targeted cancer chemotherapy.