Muscle velocity recovery cycles in myopathy.

OBJECTIVE: To understand the pathophysiology of myopathies by using muscle velocity recovery cycles (MVRC) and frequency ramp (RAMP) methodologies. METHODS: 42 patients with quantitative electromyography (qEMG) and biopsy or genetic verified myopathy and 42 healthy controls were examined with qEMG, MVRC and RAMP, all recorded from the anterior tibial muscle. RESULTS: There were significant differences in the motor unit potential (MUP) duration, the early and late supernormalities of the MVRC and the RAMP latencies in myopathy patients compared to controls (p < 0.05 apart from muscle relatively refractory period (MRRP)). When dividing into subgroups, the above-mentioned changes in MVRC and RAMP parameters were increased for the patients with non-inflammatory myopathy, while there were no significant changes in the group of patients with inflammatory myopathy. CONCLUSIONS: The MVRC and RAMP parameters can discriminate between healthy controls and myopathy patients, more significantly for non-inflammatory myopathy. MVRC differences with normal MRRP in myopathy differs from other conditions with membrane depolarisation. SIGNIFICANCE: MVCR and RAMP may have a potential in understanding disease pathophysiology in myopathies. The pathogenesis in non-inflammatory myopathy does not seem to be caused by a depolarisation of the resting membrane potential but rather by the change in sodium channels of the muscle membrane.

Muscle velocity recovery cycles in neurogenic muscles.

OBJECTIVE: To examine muscle membrane properties in neurogenic muscles using Muscle Velocity Recovery Cycles (MVRCs). METHODS: Forty-seven patients referred to Nerve Conduction Studies (NCS) and Electromyography (EMG) for peroneal nerve entrapment neuropathy were prospectively included. The patients were categorized as peroneal nerve entrapment neuropathy across knee (n = 22), L5-radiculapathy (n = 10), normal NCS/EMG (n = 9) and other disorders (n = 6) using NCS/EMG and neuroimaging results. Strength in anterior tibial muscle was measured by Medical Council Scale (MRC) and disease duration was recorded. In addition to conventional NCS/EMG, all subjects were examined with MVRCs in anterior tibial muscle. This provided parameters of muscle relative refractory period (MRRP) and early supernormality (ESN) and late supernormality (LSN). The results were compared with 29 age-matched healthy control subjects. RESULTS: MRRP was prolonged and ESN and LSN were reduced in neurogenic muscles. MRRP, ESN and LSN correlated to MRC and incidence of spontaneous activity but not to motor unit potential parameters or disease duration. CONCLUSIONS: MVRC changes provide in vivo evidence of depolarization in intact human muscle fibres that could underlie reduced muscle excitability and hence weakness in neurogenic muscles. SIGNIFICANCE: MVRCs appear to be a useful technique for revealing disease mechanism in a broad range of neuromuscular diseases.

MScanFit motor unit number estimation (MScan) and muscle velocity recovery cycle recordings in amyotrophic lateral sclerosis patients.

OBJECTIVE: Motor Unit Number Estimation (MUNE) methods, such as the recently developed MScanFit MUNE (MScan), may be valuable in tracking motor unit loss in ALS. Muscle Velocity Recovery Cycles (MVRCs) provide information about muscle membrane properties and can reveal disease-related changes. This study was undertaken to test the applicability of MScan to the anterior tibial muscle (TA) and to test whether the MVRCs could improve understanding of ALS pathophysiology. METHODS: Twenty-six ALS patients and 25 healthy controls were evaluated by quantitative electromyography, nerve conduction study and the two novel methods: MScan and MVRC; all in the TA and peroneal nerve. RESULTS: The estimated number of motor units for ALS patients (Median: 45, interquartile range: 28.5-76.5) was significantly lower than for the controls (117, 96.0-121.0) (P = 2.19 × 10-7). Unit size was increased only when amplitudes were expressed as percentage of CMAP. Of MVRC measurements, only relative refractory period was significantly abnormal in patients. CONCLUSION: MScanFit MUNE gives a sensitive and quantitative measure of loss of TA motor units in ALS. Muscle fiber membrane properties are mostly unaffected, despite substantial denervation, presumably due to collateral reinnervation. SIGNIFICANCE: MScan is suitable for detecting motor unit loss in TA. MVRCs do not provide new insights in ALS.

The utility of motor unit number estimation methods versus quantitative motor unit potential analysis in diagnosis of ALS.

OBJECTIVE: To compare the diagnostic utility of motor unit number estimation (MUNE) methods to motor unit potential (MUP) analysis in amyotrophic lateral sclerosis (ALS). METHODS: Twenty-five patients (1 definite, 11 probable, 9 possible ALS and 4 progressive muscular atrophy) and 22 healthy controls were prospectively included. Quantitative MUP analysis and three MUNE methods; Multiple Point Stimulation MUNE (MPS), Motor Unit Number Index (MUNIX) and MScanFit MUNE (MScan) were done in abductor pollicis brevis muscle. The sensitivities were compared by McNemar chi-square test. MUNE, MUP and revised ALS Functional Rating Scale (ALSFRS-R) parameters were correlated by regression analysis. RESULTS: The sensitivities of MPS (76%) and MScan (68%) were higher than MUP duration (36%) and amplitude (40%) in detecting motor unit loss (p < 0.05). MUNE methods increased the categorical probability from possible to probable ALS in 4 patients (16%). There was only significant correlation between ALSFRS-R and MScan (r = 0.443, p = 0.027) among the electrophysiological tests. MUNE methods did not correlate to MUP parameters. CONCLUSIONS: MUNE methods are more sensitive in showing abnormality than MUP analysis. SIGNIFICANCE: MUNE methods, in particular MScan, may have the potential to be implemented in the clinical practice for diagnosis and follow-up of neuromuscular disorders particularly ALS.

Prevalence, antimicrobial resistance, and molecular characterization of Campylobacter spp. in bulk tank milk and milk filters from US dairies.

Campylobacter spp. are frequently isolated from dairy cows as commensal organisms. Sporadic Campylobacter infections in humans in the United States are generally attributed to poultry, but outbreaks are also commonly associated with dairy products, particularly unpasteurized or raw milk. Bulk tank milk samples and milk filters from US dairy operations were collected during the National Animal Health Monitoring System Dairy 2014 study and analyzed using real-time PCR and traditional culture techniques for the presence of thermophilic Campylobacter species. The weighted prevalence of operations from which we detected Campylobacter spp. in either bulk tank milk or milk filters was 24.9%. We detected Campylobacter spp. in a higher percentage of operations with 100-499 cows (42.8%) and 500 or more cows (47.5%) than in operations with 30-99 cows (6.5%). Campylobacter spp. were also more frequently detected in operations in the west than the east (45.9 and 22.6%, respectively). We isolated Campylobacter spp. from approximately half of PCR-positive samples, representing 12.5% (weighted prevalence) of operations. The majority (91.8%) of isolates were C. jejuni, but C. lari and C. coli were also isolated. We detected resistance to tetracycline in 68.4% of C. jejuni isolates, and resistance to ciprofloxacin and nalidixic acid in 13.2% of C. jejuni isolates. Based on pulsed-field gel electrophoresis, we found that dairy-associated C. jejuni were genotypically diverse, although clonal strains were isolated from different geographic regions. These results suggest that bulk tank milk can be contaminated with pathogenic Campylobacter spp., and that the consumption of unpasteurized or raw milk presents a potential human health risk.

Saprolegnia species in Norwegian salmon hatcheries: field survey identifies S. diclina sub-clade IIIB as the dominating taxon.

Saprolegnia isolates within the recognized clades encompassing the taxa S. parasitica and S. diclina act as opportunist and aggressive pathogens to both fish and their eggs. They are responsible for significant economic losses in aquaculture, particularly in salmonid hatcheries. However, the identity, distribution and pathogenic significance of involved species often remain unexplored. In this study, 89 Saprolegnia isolates were recovered from water, eggs and salmon tissue samples that originated from salmon (Salmo salar) hatcheries along the coast of Norway. The cultures were characterized morphologically and molecularly in order to provide an overview of the species composition of Saprolegnia spp. present in Norwegian salmon hatcheries. We demonstrate that S. diclina clearly dominated and contributed to 79% of the recovered isolates. Parsimony analyses of the nuclear ribosomal internal transcribed spacer (ITS) region split these isolates into 2 strongly supported sub-clades, S. diclina sub-clade IIIA and IIIB, where sub-clade IIIB accounted for 66% of all isolates. A minor portion of the isolates constituted other taxa that were either conspecific or showed strong affinity to S. parasitica, S. ferax, S. hypogyna and Scoliolegnia asterophora. The unique sub-clade IIIB of S. diclina was most prevalent in water and salmon eggs, while S. parasitica isolates were more frequently isolated from post hatching stages. The study demonstrated that morphological criteria in many cases were insufficient for species delimitation due to lack of sexual structures or incoherent morphological expression of such features within the tested replicates.

Facial nerve palsy caused by parotid gland abscess.

OBJECTIVES: We present the first report of methicillin-resistant Staphylococcus aureus and Propionibacterium acnes parotid abscesses complicated by facial nerve palsy. Facial nerve palsy secondary to parotid gland abscess is rare, with only eight previously reported cases. METHOD: Case reports and literature review concerning parotid abscess and facial nerve palsy presentation and management. CASE REPORTS: Within two months, two female patients presented with parotid gland abscess complicated by unilateral facial paralysis. Both were treated with intravenous antibiotics and surgery. In the first case, methicillin-resistant Staphylococcus aureus was cultivated, in the other, Propionibacterium acnes was found. In the first case, facial nerve function did not recover. CONCLUSION: Parotid gland abscess can lead to facial paralysis. Both methicillin-resistant Staphylococcus aureus and Propionibacterium acnes may be involved. Ultrasonography or computed tomography is recommended to exclude a parotid abscess in patients presenting with suppurative parotitis.

Parental stress and marital relationships among patients with brain injury and their spouses.

INTRODUCTION/OBJECTIVES: This study examines parental stress and marital relationship among patients with acquired brain injury and their spouses. PARTICIPANTS, MATERIALS/METHODS: The participants were 35 patients diagnosed with acquired brain injury and having school-aged children, together with their spouses recruited from out-patients from brain injury rehabilitation units across Denmark. The parents self-reported parental stress using the Parental Stress Index (PSI), marital adjustment using the Dyadic Adjustment Scale (DAS), family functioning using the Family Environment Scale (FES). Additionally, the healthy parents completed the Symptom Checklist 90 (SCL-90) measuring psychological symptoms. A matched control group consisted of parents suffering from diabetes and their spouses, being recruited from the National Danish Diabetes Register. RESULTS: Significantly more symptoms of parental stress were reported by the brain-injured parents when compared to the diabetic parents (p < 0.05); the brain injured parents also reported significant more family dysfunction (p < 0.05). The healthy parents in the brain injury group rated their marital satisfaction and current relationship happiness significantly lower (p < 0.05) than the healthy control group. The spouses to the brain-injured patients were also significantly more depressed (p < 0.01). CONCLUSIONS: When parents have acquired brain injury, they experience more parental stress and family dysfunction. Their spouses are less satisfied in their marital relationship and feel more depressed. These results indicate the need of a family centred supportive service to focus on the relational changes and dysfunctional family processes in families with a brain-injured parent and dependent children.

Survival in extreme environments - on the current knowledge of adaptations in tardigrades.

Tardigrades are microscopic animals found worldwide in aquatic as well as terrestrial ecosystems. They belong to the invertebrate superclade Ecdysozoa, as do the two major invertebrate model organisms: Caenorhabditis elegans and Drosophila melanogaster. We present a brief description of the tardigrades and highlight species that are currently used as models for physiological and molecular investigations. Tardigrades are uniquely adapted to a range of environmental extremes. Cryptobiosis, currently referred to as a reversible ametabolic state induced by e.g. desiccation, is common especially among limno-terrestrial species. It has been shown that the entry and exit of cryptobiosis may involve synthesis of bioprotectants in the form of selective carbohydrates and proteins as well as high levels of antioxidant enzymes and other free radical scavengers. However, at present a general scheme of mechanisms explaining this phenomenon is lacking. Importantly, recent research has shown that tardigrades even in their active states may be extremely tolerant to environmental stress, handling extreme levels of ionizing radiation, large fluctuation in external salinity and avoiding freezing by supercooling to below -20 °C, presumably relying on efficient DNA repair mechanisms and osmoregulation. This review summarizes the current knowledge on adaptations found among tardigrades, and presents new data on tardigrade cell numbers and osmoregulation.

DNA microarray to detect and identify trichothecene- and moniliformin-producing Fusarium species.

AIMS: To develop a DNA microarray for easy and fast detection of trichothecene- and moniliformin-producing Fusarium species. METHOD AND RESULTS: A DNA microarray was developed for detection and identification of 14 trichothecene- and moniliformin-producing species of the fungal genus Fusarium. The array could also differentiate between four species groups. Capture probes were designed based on recent phylogenetic analyses of translation elongation factor-1 alpha (TEF-1alpha) sequences. Particular emphasis was put on designing capture probes corresponding to groups or species with particular mycotoxigenic synthetic abilities. A consensus PCR amplification of a part of the TEF-1alpha is followed by hybridization to the Fusarium chip and the results are visualized by a colorimetric Silverquant detection method. We validated the Fusarium chip against five naturally infected cereal samples for which we also have morphological and chemical data. The limit of detection was estimated to be less than 16 copies of genomic DNA in spiked commercial wheat flour. CONCLUSIONS: The current Fusarium chip proved to be a highly sensitive and fast microarray for detection and identification of Fusarium species. We postulate that the method also has potential for (semi-)quantification. SIGNIFICANCE AND IMPACT OF THE STUDY: The Fusarium chip may prove to be a very valuable tool for screening of cereal samples in the food and feed production chain, and may facilitate detection of new or introduced Fusarium spp.

Simultaneous detection and identification of trichothecene- and moniliformin-producing Fusarium species based on multiplex SNP analysis.

AIM: To develop a multiplex identification method for trichothecene- and moniliformin-producing Fusarium species. METHOD AND RESULTS: In this article, we present a single nucleotide polymorphism (SNP) assay to simultaneously detect and identify 16 trichothecene- and moniliformin-producing Fusarium species. A number of SNP primers are designed to detect clades of species with particular mycotoxigenic synthetic abilities. The assay is based on minisequencing using SNaPshot reactions and the SNP primers are designed based on motifs derived from phylogenetic analyses of translation elongation factor-1alpha sequences. The present version of the Fusarium SNP assay can distinguish major groups of trichothecene producers; the strict-type-A, the strict-type-B, the type-A and type-B trichothecene producers and the putative moniliformin producers. The SNP assay was validated against five naturally infected cereal samples that previously had been analysed morphologically, chemically and by a multiplex DNA array hybridization. CONCLUSIONS: The Fusarium SNP assay reveals the advantages of using SNPs for multiplex species identification. SIGNIFICANCE AND IMPACT OF THE STUDY: The current assay may qualify as a high-throughput screening method for small-grain cereals in the feed and food chain, and may facilitate detection of new or introduced Fusarium species.

Genetic structure and evolution of RAC-GTPases in Arabidopsis thaliana.

Rho GTPases regulate a number of important cellular functions in eukaryotes, such as organization of the cytoskeleton, stress-induced signal transduction, cell death, cell growth, and differentiation. We have conducted an extensive screening, characterization, and analysis of genes belonging to the Ras superfamily of GTPases in land plants (embryophyta) and found that the Rho family is composed mainly of proteins with homology to RAC-like proteins in terrestrial plants. Here we present the genomic and cDNA sequences of the RAC gene family from the plant Arabidopsis thaliana. On the basis of amino acid alignments and genomic structure comparison of the corresponding genes, the 11 encoded AtRAC proteins can be divided into two distinct groups of which one group apparently has evolved only in vascular plants. Our phylogenetic analysis suggests that the plant RAC genes underwent a rapid evolution and diversification prior to the emergence of the embryophyta, creating a group that is distinct from rac/cdc42 genes in other eukaryotes. In embryophyta, RAC genes have later undergone an expansion through numerous large gene duplications. Five of these RAC duplications in Arabidopsis thaliana are reported here. We also present an hypothesis suggesting that the characteristic RAC proteins in higher plants have evolved to compensate the loss of RAS proteins.

Micrognathozoa: a new class with complicated jaws like those of Rotifera and Gnathostomulida.

A new microscopic aschelminth-like animal, Limnognathia maerski nov. gen. et sp., is described from a cold spring at Disko Island, West Greenland, and assigned to Micrognathozoa nov. class. It has a complex of jaws in its pharynx, and the ultrastructure of the main jaws is similar to that of the jaws of advanced scleroperalian gnathostomulids. However, other jaw elements appear also to have characteristics of the trophi of Rotifera. Jaw-like structures are found in other protostome taxa as well-for instance, in proboscises of kalyptorhynch platyhelminths, in dorvilleid polychaetes and aplacophoran mollusks-but studies of their ultrastructure show that none of these jaws is homologous with jaws found in Gnathostomulida, Rotifera, and Micrognathozoa. The latter three groups have recently been joined into the monophylum Gnathifera Ahlrichs, 1995, an interpretation supported by the presence of jaw elements with cuticular rods with osmiophilic cores in all three groups. Such tubular structures are found in the fulcrum of all Rotifera and in several cuticular sclerites of both Gnathostomulida and Micrognathozoa. The gross morphology of the pharyngeal apparatus is similar in the three groups. It consists of a ventral pharyngeal bulb and a dorsal pharyngeal lumen. The absence of pharyngeal ciliation cannot be used as an autapomorphy in the ground pattern of the Gnathifera because the Micrognathozoa has the plesiomorphic alternative with a ciliated pharyngeal epithelium. The body of Limnognathia maerski nov. gen. et sp. consists of a head, thorax, and abdomen. The dorsal and lateral epidermis have plates formed by an intracellular matrix, as in Rotifera and Acanthocephala; however, the epidermis is not syncytial. The ventral epidermis lacks internal plates, but has a cuticular oral plate without ciliary structures. Two ventral rows of multiciliated cells form a locomotory organ. These ciliated cells resemble the ciliophores present in some interstitial annelids. An adhesive ciliated pad is located ventrally close to a caudal plate. As in many marine interstitial animals-e.g., gnathostomulids, gastrotrichs, and polychaetes-a special form of tactile bristles or sensoria is found on the body. Two pairs of protonephridia with unicellular terminal cells are found in the trunk; this unicellular condition may be the plesiomorphic condition in Bilateria. Only specimens with the female reproductive system have been found, indicating that all adult animals are parthenogenetic females. We suggest that 1) jaws of Gnathostomulida, Rotifera, and the new taxon, Micrognathozoa, are homologous structures; 2) Rotifera (including Acanthocephala) and the new group might be sister groups, while Gnathostomulida could be the sister-group to this assemblage; and 3) the similarities to certain gastrotrichs and interstitial polychaetes are convergent.

[The requirement to use the cheapest synonymous preparation--does it work as intended?]. / Ordningen med billigste synonympreparat--virker den etter hensikten?

A retrospective analysis of all prescriptions for hypersensitive drugs was undertaken at a pharmacy serving 20 general practitioners and 17 temporarily employed doctors. The analysis was carried out in 1992 over two three-month periods separated by an interval of five months. The Governmental regulations aimed at promoting use of the "cheapest synonymous drug" apparently had little impact on the doctors' prescriptions. During the 11 months concerned, the prescribed antihypersensitive drugs contained an increasing share of expensive alternatives with no documented ability to reduce cardiovascular morbidity or mortality.

Reliability coefficients of three corneal pachymeters.

We compared the accuracy and reproducibility of a hand-held portable ultrasound pachymeter, the Pach-Pen (Bio-Rad, Ophthalmic Division, Santa Ana, California); another ultrasound pachymeter, the DGH 1000 (DGH Technology, Inc., Frazer, Pennsylvania); and the Pro-Cem 4 endothelial specular microscope (Alcon-Surgical, Inc., Irvine, California). Each eye of 18 healthy human subjects was examined to determine corneal thickness using the three different instruments. For each instrument, five repeated measurements were obtained at each of five corneal locations (one central, four peripheral), for a total of 25 measurements per eye. The accuracy of the two ultrasound pachymeters was tested by comparing measurements obtained on specially designed test blocks of known thickness. The Pach-Pen was the more accurate of the two ultrasound pachymeters, with measurements within the range of 0.003 to 0.065 mm from the true thickness. The three instruments were most consistent in mean thickness in the center of the cornea. All three instruments showed excellent intraobserver reproducibility, as measured by reliability coefficients over 90%. Overall, the Pach-Pen pachymeter had high reproducibility, and produced more accurate measurements than the DGH 1000 pachymeter.

A clinical evaluation of the BioPen.

We conducted a clinical trial on the Oculab BioPen, a portable, handheld applanation instrument designed to measure ocular axial lengths. We compared the measurements obtained from the BioPen with those obtained from the Ultrascan Digital B System IV from CooperVision. Accuracy and reproducibility were assessed in vitro by performing ten measurements with each instrument on a precalibrated 25.8-mm plastic test block. The in vivo reproducibility of the BioPen was evaluated by performing five serial readings on each eye of 58 patients. Keratometry measurements were also recorded to determine whether the BioPen provided consistent readings regardless of corneal curvature. We found the BioPen to be as accurate and reproducible as the Ultrascan Digital B in vitro and in vivo. Corneal curvature had no effect on the in vivo reproducibility of the BioPen.

The effectiveness of hearing protectors in practice.

A series of workmen being evaluated for pensions for occupational hearing loss were asked to bring their own hearing protectors from work, to fit them themselves, following which attenuation studies were made. The muffs and most earplugs produced similar attenuation levels at high frequencies, although the muffs produced less attenuation at low frequencies. In all cases the mean attenuation was significantly lower than optimum figures suggested in the literature, and the standard deviation was relatively high. Personally molded earplugs were significantly less effective than other plugs used. Reasons are discussed for the relatively poor performance of these devices and the concept of assumed protection, i.e. mean minus one standard deviation, is discussed. There is need for better instruction on how to use hearing protectors if they are to be effective.

Ipsilateral acoustic reflex stimulation in normal and sensorineural impaired ears: a preliminary report.

Ipsilateral acoustic reflex thresholds were measured at 1,000 Hz and 2,000 Hz in 38 subjects with normal hearing and in 107 subjects with a symmetrical sensorineural hearing loss. Acoustic reflex thresholds were approximately six dB better for ipsilateral vs. contralateral stimuli for both subject groups. High ipsilateral reflex thresholds were associated with larger static compliance values in the sensorineural hearing impaired subjects. Reversed reflexes (increasing compliance with muscle contraction) were observed for ipsilateral stimulation in five per cent (two) of the normal hearing subjects, and 23 percent (25) of the sensorineural hearing impaired subjects. The number of reversed reflexes was greater for subjects with higher hearing thresholds and higher ipsilateral acoustic reflex thresholds. The results suggest that quantitative use of the ipsilateral acoustic reflex requires additional research. At present it seems prudent to use ipsilateral acoustic reflex measurements as a qualitative rather than a quantitative tool, e.g. in confirming the status of one middle ear when the other has a conductive hearing loss. However, even this limited role has a considerable clinical potential.