Estimating motor unit numbers from a CMAP scan: Repeatability study on three muscles at 15 centres.

OBJECTIVE: To assess the repeatability and suitability for multicentre studies of MScanFit motor unit number estimation (MUNE), which involves modelling compound muscle action potential (CMAP) scans. METHODS: Fifteen groups in 9 countries recorded CMAP scans twice, 1-2 weeks apart in healthy subjects from abductor pollicis brevis (APB), abductor digiti minimi (ADM) and tibialis anterior (TA) muscles. The original MScanFit program (MScanFit-1) was compared with a revised version (MScanFit-2), designed to accommodate different muscles and recording conditions by setting the minimal motor unit size as a function of maximum CMAP. RESULTS: Complete sets of 6 recordings were obtained from 148 subjects. CMAP amplitudes differed significantly between centres for all muscles, and the same was true for MScanFit-1 MUNE. With MScanFit-2, MUNE differed less between centres but remained significantly different for APB. Coefficients of variation between repeats were 18.0% for ADM, 16.8% for APB, and 12.1% for TA. CONCLUSIONS: It is recommended for multicentre studies to use MScanFit-2 for analysis. TA provided the least variable MUNE values between subjects and the most repeatable within subjects. SIGNIFICANCE: MScanFit was primarily devised to model the discontinuities in CMAP scans in patients and is less suitable for healthy subjects with smooth scans.

[Unusual Dependence between Gene Expression and Negative Selection in Euplotes].

In most of the studied organisms, gene expression is associated with a number of evolutionary features pertaining to the protein-coding sequences. In particular, gene expression positively correlates with the average intensity of negative selection and influences codon usage. Here, we study the connection between gene expression and selection patterns in two species of ciliate protists of the genus Euplotes. We find that codon usage is influenced by gene expression in these organisms, pointing at additional evolutionary constraints on mutations in heavily expressed genes relative to the genes expressed at lower rates. At the same time, at the level of synonymous vs. non-synonymous substitutions we observe a stronger constraint on the genes expressed at lower rates relative to those with higher rates of expression. Our study adds to the discussion about the general evolutionary patterns and opens new questions about the mechanisms of control of gene expression in ciliates.

Raman spectroscopy in oral cavity and oropharyngeal cancer: a systematic review.

Due to the high incidence of head and neck cancer and under-diagnosis in the early stages, non-invasive and highly accurate diagnostic tests are required for cancer detection. Recent advances in Raman spectroscopy techniques have yielded promising sensitivity and specificity results in the evaluation of cancer. The aim of this study was to investigate the potential value of Raman spectroscopy in oral cavity and oropharyngeal cancer diagnosis based on currently available scientific papers. A search of the PubMed database was performed using a specific strategy and according to the PRISMA guidelines. Raman spectroscopy achieved a maximum accuracy of 98% in cancer detection, while accuracy was 97.24% for tumour grading evaluation, 95% for cancer treatment assessment, and 77% for the detection of cancer recurrence. Moreover, early-stage cancer can be identified by Raman spectroscopy investigation of liquid biopsy samples. An in vivo technique with direct mucosa examination by fibre-optic Raman spectroscopy obtained a maximum accuracy of 94% in cancer diagnosis. The most prominent markers of the presence of malignancy were an increase in Raman signal intensity for proteins, nucleic acids, and water and a decrease for lipids. These cancer discriminants were detected in both fingerprint and high wavenumber regions. In conclusion, Raman spectroscopy is a promising tool for oral cavity and oropharyngeal cancer screening.

Comprehensive survey on radon mitigation and indoor air quality in energy efficient buildings from Romania.

Over the last 10 years applied scientific research has been carried out in Romania to tacked the residential radon issues. The increased interest to reduce the carbon footprint of buildings has lead to the implementation and use of new architectural solutions aimed to save energy in houses and other buildings. As a consequence, the degree of retrofit in existing buildings and energy efficiency of new buildings promoted the need to not only mitigate indoor radon, but improve indoor air quality overall. The present study found that the while the best performance in radon reduction was confirmed to be based on sub-slab depressurization (61% - 95% reduction), centralized and decentralized mechanical supply and exhaust ventilation with heat recovery yielded a good efficiency in overall improvement of indoor air quality (CO2, VOC, RH, temperature). The outcome of our research, as well as future perspectives, take into account the recommended harmonization of energy efficiency programs with those of public health by finding and applying the best technologies in compliance with energy saving and indoor environmental quality.

Increased Axon Initial Segment Length Results in Increased Na+ Currents in Spinal Motoneurones at Symptom Onset in the G127X SOD1 Mouse Model of Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease preferentially affecting motoneurones. Transgenic mouse models have been used to investigate the role of abnormal motoneurone excitability in this disease. Whilst an increased excitability has repeatedly been demonstrated in vitro in neonatal and embryonic preparations from SOD1 mouse models, the results from the only studies to record in vivo from spinal motoneurones in adult SOD1 models have produced conflicting findings. Deficits in repetitive firing have been reported in G93A SOD1(high copy number) mice but not in presymptomatic G127X SOD1 mice despite shorter motoneurone axon initial segments (AISs) in these mice. These discrepancies may be due to the earlier disease onset and prolonged disease progression in G93A SOD1 mice with recordings potentially performed at a later sub-clinical stage of the disease in this mouse. To test this, and to explore how the evolution of excitability changes with symptom onset we performed in vivo intracellular recording and AIS labelling in G127X SOD1 mice immediately after symptom onset. No reductions in repetitive firing were observed showing that this is not a common feature across all ALS models. Immunohistochemistry for the Na+ channel Nav1.6 showed that motoneurone AISs increase in length in G127X SOD1 mice at symptom onset. Consistent with this, the rate of rise of AIS components of antidromic action potentials were significantly faster confirming that this increase in length represents an increase in AIS Na+ channels occurring at symptom onset in this model.

Shorter axon initial segments do not cause repetitive firing impairments in the adult presymptomatic G127X SOD-1 Amyotrophic Lateral Sclerosis mouse.

Increases in axonal sodium currents in peripheral nerves are some of the earliest excitability changes observed in Amyotrophic Lateral Sclerosis (ALS) patients. Nothing is known, however, about axonal sodium channels more proximally, particularly at the action potential initiating region - the axon initial segment (AIS). Immunohistochemistry for Nav1.6 sodium channels was used to investigate parameters of AISs of spinal motoneurones in the G127X SOD1 mouse model of ALS in adult mice at presymptomatic time points (~190 days old). In vivo intracellular recordings from lumbar spinal motoneurones were used to determine the consequences of any AIS changes. AISs of both alpha and gamma motoneurones were found to be significantly shorter (by 6.6% and 11.8% respectively) in G127X mice as well as being wider by 9.8% (alpha motoneurones). Measurements from 20-23 day old mice confirmed that this represented a change during adulthood. Intracellular recordings from motoneurones in presymptomatic adult mice, however, revealed no differences in individual action potentials or the cells ability to initiate repetitive action potentials. To conclude, despite changes in AIS geometry, no evidence was found for reduced excitability within the functional working range of firing frequencies of motoneurones in this model of ALS.

[Prokaryotic and Mitochondrial Linear Genomes: Their Genesis, Evolutionary Significance, and the Problem of Replicating Chromosome Ends].

Bacterial chromosomes are widely thought of as circular DNA molecules. However, linear bacterial chromosomes, as well as linear mitochondrial and plastid chromosomes, are fairly common. The most frequent causes of linearization are reparation system defects, incorporation of plasmids in the genome, and recombination compromising the circular topology of chromosomes. Genomes of some bacterial species had undergone frequent linearization-circularization events, which resulted in an increased variability of gene content at linear chromosome ends. Similarly to eukaryotes, bacteria that have linear genomes face the problem of end replication, which different species solve in a variety of ways. A theoretically important issue is the adaptive value of chromosome linearization. This review discusses theories concerning the evolution of linear genomes and supporting experiments. The most common mechanisms of linear bacterial genomes replication and possible ways of their emergence are also considered.

The FIRST large-scale mapping of radon concentration in soil gas and water in Romania.

In the framework of the last Council Directive 2013/59 (Euratom, 2014) laying down basic safety standards for protection against the dangers arising from exposure to ionizing radiation, the problem of radon was assumed in Romania at national level by responsible authorities through the design and development of a National Radon Action Plan and an adequate legislation (HG nr. 526/2018). In order to identify radon risk areas, however, it is necessary to perform systematic radon measurements in different environmental media (soil gas, water, indoor air) and to map the results. This paper presents an atlas of up-to-date radon in soil and water levels for central and western part of Romania. The radon in soil map includes data from 2564 measurements carried out on-site, using Luk3C radon detector. The Luk-VR system was used to measure radon activity concentration from 2452 samples of drinking water. The average radon activity concentration was 29.3 kBq m-3 for soil gas, respectively 9.8 Bq l-1 for water dissolved air. Mapping of radon can be a useful tool to implement radon policies at both the national and local levels, defining priority areas for further study when land-use decisions must be made.

Skin wound regeneration with bioactive glass-gold nanoparticles ointment.

The bioactive glasses can lead to the promotion of growth of granulation tissue, while the gold nanoparticles (AuNPs) can induce the acceleration of wound healing including tissue regeneration, connective tissue formation, and angiogenesis. The aim of this study was to evaluate the impact of using the bioactive glass (BG) and BG-AuNPs composites on skin wound healing in experimental rat models for 14 days. Sol-gel derived BGs and BG-AuNPs composites mixed with Vaseline at 6, 12 and 18 wt% were used to evaluate the repair response of the skin. During the process of healing, granulomatous reaction was observed in the wound treated with 12 and 18 wt% BG-Vaseline ointments. Furthermore, a strong vascular proliferation and complete wound regeneration were found in 18%BG-AuNPs-Vaseline treated groups. The results derived from the performed investigations revealed that the 18% BG-AuNPs-Vaseline ointment is a promising candidate for wound healing applications.

Applications of inflammation-derived gingival stem cells for testing the biocompatibility of dental restorative biomaterials.

BACKGROUND: Normal or inflamed gingival tissues are regarded as a source of mesenchymal stem cells (MSCs) abundant and easily accessible through minimally invasive dental procedures. Due to the proximity of dental resin composites to gingival tissues and to the possible local cytotoxic effect of the eluted components, gingiva-derived MSCs could be used to investigate the biocompatibility of dental biomaterials. PURPOSE: The present research aimed to isolate (MSCs) from inflamed and normal gingiva, to fully characterize them and to observe their behavior in relation with some commercial resin composite materials and one experimental material. MATERIAL AND METHODS: Following their isolation, putative MSCs from both gingival sources were grown under the same culture conditions and characterized by immunophenotyping of cell surface antigens by flow-cytometry and transcription factors by immunocytochemical staining. Moreover, stemness gene expression was evaluated by RT-PCR analysis. Multipotent mesenchymal differentiation potential was investigated. Osteogenic and neurogenic differentiated cells were highlighted by immunocytochemical staining, chondrogenic cells by cytochemical staining, and adipocytes by cytochemical staining and spectrophotometry, respectively. Resin composite cytotoxicity was evaluated by cell membrane fluorescent labeling with PKH 26 and MTT assay. The results of PKH labeling were statistically analysed using two-way RM ANOVA with Bonferroni post-tests. For MTT assay, two-way RM ANOVA with Bonferroni post-tests and unpaired t test with Welch's correction were used. RESULTS: A similar expression pattern of surface markers was observed. The cells were positive for CD105, CD73, CD90, CD49e, CD29, CD44 and CD166 and negative for CD45, CD34, CD14, CD79, HLA-DR and CD117 indicating a mesenchymal stem cell phenotype. The qRT-PCR analysis revealed a low gene expression for NOG, BMP4 and Oct3/4 and an increased expression for Nanog in both cells lines. Immunocytochemical analysis highlighted a more intense protein expression for Nanog, Oct3/4 and Sox-2 in MSCs derived from normal gingiva than from inflamed gingiva. Multipotent differentiation capacity of MSCs isolated from both sources was highlighted. The tested materials had no hazardous effect on MSCs as the two cell lines developed well onto resin composite substrates. Cell counting revealed some significant differences in the number of PKH-labeled MSCs at some experimental moments. Also, some differences in cell viability were recorded indicating better developmental conditions offered by some of the tested biomaterials. CONCLUSIONS: The experimental resin composite behaved like the most biocompatible commercial material. Inflamed gingiva-derived MSCs retain their stem cell properties and could be used as a valuable cell line for testing dental biomaterials.

THE GEOGENIC RADON POTENTIAL MAP OF THE ASPIRING 'BUZAU LAND' GEOPARK.

Mapping the geogenic radon potential in Buzau County is part of a research project aiming to apply research for sustainable development and economic growth following the principles of geoconservation in order to support the 'Buzau Land' UNESCO Geopark initiative. The mapping of geogenic radon will be used as an overview for planning purposes. The main geological formations of the studied area were identified as Cretaceous and Paleogene flysch, included in a thin-skinned nappes pile and consisting of alternating sandstones, marls, clays and, subordinately, conglomerates, all tightly folded or faulted. Significant variations in the concentration of radon were therefore determined in the ground. However, no high values were determined, the maximum measured activity concentration being 101.6 kBq m-3.

Response to interferon-beta treatment in multiple sclerosis patients: a genome-wide association study.

Up to 50% of multiple sclerosis (MS) patients do not respond to interferon-beta (IFN-ß) treatment and determination of response requires lengthy clinical follow-up of up to 2 years. Response predictive genetic markers would significantly improve disease management. We aimed to identify IFN-ß treatment response genetic marker(s) by performing a two-stage genome-wide association study (GWAS). The GWAS was carried out using data from 151 Australian MS patients from the ANZgene/WTCCC2 MS susceptibility GWAS (responder (R)=51, intermediate responders=24 and non-responders (NR)=76). Of the single-nucleotide polymorphisms (SNP) that were validated in an independent group of 479 IFN-ß-treated MS patients from Australia, Spain and Italy (R=273 and NR=206), eight showed evidence of association with treatment response. Among the replicated associations, the strongest was observed for FHIT (Fragile Histidine Triad; combined P-value 6.74 × 10-6) and followed by variants in GAPVD1 (GTPase activating protein and VPS9 domains 1; combined P-value 5.83 × 10-5) and near ZNF697 (combined P-value 8.15 × 10-5).

The influence of three endodontic sealers on bone healing: an experimental study.

BACKGROUND: The aim of this experimental study is to assess the bone healing phenomenon produced in the presence of 3 dental materials used for the root canal obturation. MATERIALS AND METHODS: The biocompatibility of 3 endodontic sealers (a self-curing epoxy resin - AH Plus, a dual cure urethane dimethacrylate resin - RealSeal and a new dual cure endodontic hydroxyapatite based filling material) was investigated after intra-osseous implantation of the materials in rats' calvaria. Tissue reaction was studied at 2, 4, 6, 8, 10 and 12 weeks after implantation using calibrated image retrieval by Olympus. We took into consideration the presence of inflammatory cells (polymorphonuclear leukocytes, macrophages, plasma cells, lymphocytes and giant cells) and classified the aspects of the histological samples according to the following scale: 0 - no inflammation, 1 - mild, isolated inflammation, 2 - moderate, localised inflammatory reaction, 3 - severe, diffuse and intense inflammatory reaction. RESULTS: The inflammatory reaction was present at the 6 intervals for all the tested materials, but a decrease of the inflammatory infiltrate, statistically significant, until extinction for all tested materials was observed at the end of the experimental period. The reaction of bone tissue recovery was most intense in the case of the control area. Lower intensity osteogenesis phenomenon was observed in case of all 3 tested sealers at the end of the experimental period. CONCLUSIONS: Biocompatibility and bone healing induction of the new hydroxyapatite based endodontic filling material is comparable to other commercial materials, AH Plus and RealSeal.

Vitamin D fortification and seasonality of birth in type 1 diabetic cases: D-tect study.

Fortification of margarine with vitamin D was mandatory in Denmark during 1961-1985. The aim of the study was to assess whether gestational and early infancy exposure to margarine fortification was associated with seasonality of birth in Danish type 1 diabetes (T1D) patients. The risks of T1D in Danes born during various exposure periods around margarine fortification termination in 1985 were analyzed. As expected, the T1D hazards in males unexposed to margarine fortification and born in spring were higher than in males born in autumn: relevant hazard ratios (95% confidence intervals) in various exposure groups ranged from 1.74 (1.112/2.708) to 37.43 (1.804/776.558). There were no indications of seasonality of birth in males exposed to fortification, nor in both exposed and unexposed females. The study suggests that early life exposure to low-dose vitamin D from fortified food eliminates seasonality of birth in T1D male patients. Further studies are required to investigate the identified gender differences.

Postactivation depression of the Ia EPSP in motoneurons is reduced in both the G127X SOD1 model of amyotrophic lateral sclerosis and in aged mice.

Postactivation depression (PActD) of Ia afferent excitatory postsynaptic potentials (EPSPs) in spinal motoneurons results in a long-lasting depression of the stretch reflex. This phenomenon (PActD) is of clinical interest as it has been shown to be reduced in a number of spastic disorders. Using in vivo intracellular recordings of Ia EPSPs in adult mice, we demonstrate that PActD in adult (100-220 days old) C57BL/6J mice is both qualitatively and quantitatively similar to that which has been observed in larger animals with respect to both the magnitude (with ∼20% depression of EPSPs at 0.5 ms after a train of stimuli) and the time course (returning to almost normal amplitudes by 5 ms after the train). This validates the use of mouse models to study PActD. Changes in such excitatory inputs to spinal motoneurons may have important implications for hyperreflexia and/or glutamate-induced excitotoxicity in the neurodegenerative disease amyotrophic lateral sclerosis (ALS). With the use of the G127X SOD1 mutant mouse, an ALS model with a prolonged asymptomatic phase and fulminant symptom onset, we observed that PActD is significantly reduced at both presymptomatic (16% depression) and symptomatic (17.3% depression) time points compared with aged-matched controls (22.4% depression). The PActD reduction was not markedly altered by symptom onset. Comparing these PActD changes at the EPSP with the known effect of the depression on the monosynaptic reflex, we conclude that this is likely to have a much larger effect on the reflex itself (a 20-40% difference). Nevertheless, it should also be accounted that in aged (580 day old) C57BL/6J mice there was also a reduction in PActD although, aging is not usually associated with spasticity.

Radon and radium concentration in water from North-West of Romania and the estimated doses.

In the present study, the measurements of radon were carried out using the LUK-VR system based on radon gas measurements with Lucas cells. The radium concentration in water was determined, with the same device, immediately after was established the radon equilibrium with radium. The results presented here are from a survey carried out in the N-W region of Transylvania (Romania) in which were investigated the radon concentrations in natural (spring, well and surface) and drinking (tap) waters. The results showed radon concentrations within the range of 0.4-187.3 Bq l(-1) with an average value of 15.9 Bq l(-1) whereas radium concentration varied between 0.05 and 0.825 Bq l(-1) with an average value of 0.087 Bq l(-1) for all types of water covered within this survey. The corresponding annual effective ingestion dose due to radon and radium from water was determined from drinking water used by the population inhabiting the area.

Peripheral motor axons of SOD1(G127X) mutant mice are susceptible to activity-dependent degeneration.

Motor neuron disorders may be associated with mitochondrial dysfunction, and repetitive electrical impulse conduction during energy restriction has been found to cause neuronal degeneration. The aim of this study was to investigate the vulnerability of motor axons of a presymptomatic late-onset, fast-progression SOD1(G127X) mouse model of amyotrophic lateral sclerosis to long-lasting, high-frequency repetitive activity. Tibial nerves were stimulated at ankle in 7 to 8-month-old SOD1(G127X) mice when they were clinically indistinguishable from wild-type (WT) mice. The evoked compound muscle action potentials and ascending compound nerve action potentials were recorded from plantar muscles and from the sciatic nerve, respectively. Repetitive stimulation (RS) was carried out in interrupted trains of 200-Hz for 3h. During the stimulation-sequence there was progressive conduction failure in WT and, to a lesser extent, in the SOD1(G127X). By contrast, 3 days after RS the electrophysiological responses remained reduced in the SOD1(G127X) but recovered completely in WT. Additionally, morphological studies showed Wallerian degeneration in the disease model. Nerve excitability testing by "threshold-tracking" showed that axons recovering from RS had changes in excitability suggestive of membrane hyperpolarization, which was smaller in the SOD1(G127X) than in WT. Our data provide proof-of-principle that SOD1(G127X) axons are less resistant to activity-induced changes in ion-concentrations. It is possible that in SOD1(G127X) there is inadequate energy-dependent Na(+)/K(+) pumping, which may lead to a lethal Na(+) overload.

Magnetic properties of annealed core-shell CoPt nanoparticles.

A precise control and understanding of the magnetization dynamics of nanostructures is an important topic in applied nanosciences. Herein, we perform such control by annealing crystalline (Co/core)-(Pt/shell) nanoparticles. Using electron tomography, temperature dependent electron microscopy and time-resolved magneto-optics, we establish a clear correlation between the magnetization dynamics and the crystalline structure of the nanoparticles. For a mild laser annealing (370 K) the Co-Pt nanoparticles keep their core-shell structure and remain superparamagnetic with a blocking temperature T(B) = 66 K. Their time-resolved reflectivity shows that they are locally organized into a supra-crystalline ordered layer in the region of the laser spot. In contrast, a thermal annealing at higher temperatures (up to 700 K) modifies the structure of the individual nanoparticles into a CoPt crystalline ferromagnetic phase, with T(B,anneal) = 347 K. Correspondingly, the magneto-crystalline anisotropy of the annealed CoPt nanoparticles increases and their magnetization dynamics displays a motion of precession, characteristic of ferromagnetic nanostructures and which is absent in the superparamagnetic Co-Pt core-shells.

Dermatomyositis and polyradiculoneuritis, a rare association.

The association between dermatomyositis and polyradiculoneuritis is rarely citated in the speciality literature. Our patient had at first a disorder that looked like dermatomyositis, then she associated polyradiculoneuritis. At the presentation to our hospital the two diseases were combined, being very difficult to differentiate. The discrimination between these two had been made by performing a muscular biopsy and EMG studies. This emphasizes once again their importance in defining the muscular impairment from the neurogenic one.