Contraction fatigue, strength adaptations, and discomfort during conventional versus wide-pulse, high-frequency, neuromuscular electrical stimulation: a systematic review.

Neuromuscular electrical stimulation (NMES) can be delivered in a conventional form (CONVNMES) and using relatively wide-pulses and high-frequencies (WPHFNMES). WPHFNMES is proposed to reduce contraction fatigability and generate larger contractions with less discomfort than CONVNMES; however, there are no systematic reviews to guide the selection of NMES types. This systematic review compared the effects of CONVNMES versus WPHFNMES on contraction fatigability, strength adaptations, and perceived discomfort in clinical and non-clinical populations. Eight studies were included. When averaged across all non-clinical participants in individual short- and long-term studies, there was either no difference between CONVNMES and WPHFNMES for all outcomes or WPHFNMES produced more fatigability. In a subset of non-clinical participants ("responders"), however, WPHFNMES reduced contraction fatigability during a single session. Long-term studies found no differences between protocols for strength adaptations in non-clinical participants and those with multiple sclerosis. We concluded that WPHFNMES reduces contraction fatigability only in the short-term studies and in non-clinical responder participants and may exacerbate fatigability in non-responders. This review was registered in the prospective international registry of systematic reviews/PROSPERO (Registration Number: CRD42020153907). Novelty: WPHF NMES may reduce fatigue in some participants and exacerbate fatigue in others. There were no differences in long-term studies between WPHF and CONV NMES on strength adaptations. Future high-quality research is needed to optimize outcomes of NMES-based programs.

Thirteen-month duration of immunity of an oral canine vaccine against challenge with Bordetella bronchiseptica.

BACKGROUND: Very few studies have evaluated the duration of immunity of Bordetella bronchiseptica vaccines in dogs, and to date, no studies have been published on the duration of immunity of oral canine Bordetella bronchiseptica vaccines. This study was designed to determine the effectiveness of a single dose of an oral B bronchiseptica vaccine in dogs when challenged 13 months after vaccination. METHODS: Two groups of approximately eight-week-old beagles were vaccinated once with 1 ml of placebo vaccine (oral, n=17) or 1 ml of Recombitek Oral Bordetella (oral, n=17). Thirteen months after vaccination, both groups were challenged with virulent B bronchiseptica via aerosolisation. RESULTS: Thirteen of 17 dogs in the placebo group (76.5 per cent) and no dogs in the Recombitek Oral Bordetella vaccine group (0.0 per cent) developed spontaneous cough of two or more consecutive days (disease case definition). Dogs in the Recombitek Oral Bordetella group had a significantly lower prevalence of disease with prevented fraction of 1 (100 per cent prevention). In addition, the number of days coughing, duration of cough and prevalence of tracheal and nasal shedding were significantly lower for dogs vaccinated with Recombitek Oral Bordetella. CONCLUSIONS: The study demonstrated that vaccination with Recombitek Oral Bordetella is effective in preventing disease and reducing shedding 13 months after vaccination when compared with dogs vaccinated with a placebo.

Evaluation of the Effects of Synovial Multipotent Cells on Deep Digital Flexor Tendon Repair in a Large Animal Model of Intra-Synovial Tendinopathy.

Intra-synovial tendon injuries are a common orthopedic problem with limited treatment options. The synovium is a specialized connective tissue forming the inner encapsulating lining of diarthrodial joints and intra-synovial tendons. It contains multipotent mesenchymal stromal cells that render it a viable source of progenitors for tendon repair. This study evaluated the effects of autologous implantation of cells derived from normal synovium (synovial membrane cells [SMCs]) in augmenting repair in an ovine model of intra-synovial tendon injury. For this purpose, synovial biopsies were taken from the right digital flexor tendon sheath following creation of a defect to the lateral deep digital flexor tendon. Mononuclear cells were isolated by partial enzymatic digestion and assessed for MSC characteristics. Cell tracking and tendon repair were assessed by implanting 5 × 106 cells into the digital flexor tendon sheath under ultrasound guidance with the effects evaluated using magnetic resonance imaging and histopathology. Synovial biopsies yielded an average 4.0 × 105 ± 2.7 × 105 SMCs that exhibited a fibroblastic morphology, variable osteogenic, and adipogenic responses but were ubiquitously strongly chondrogenic. SMCs displayed high expression of CD29 with CD271NEGATIVE and MHC-IILOW cell-surface marker profiles, and variable expression of CD73, CD90, CD105, CD166, and MHC-I. Implanted SMCs demonstrated engraftment within the synovium, though a lack of repair of the tendon lesion over 24 weeks was observed. We conclude healthy synovium is a viable source of multipotent cells, but that the heterogeneity of synovium underlies the variability between different SMC populations, which while capable of engraftment and persistence within the synovium exhibit limited capacity of influencing tendon repair. © 2019 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society J Orthop Res 38:128-138, 2020.

Analysis of polyhexamethylene biguanide and alexidine in contact lens solutions using capillary electrophoresis, ultra-performance liquid chromatography and quadrupole time of flight mass spectrometry.

Polymeric biguanides, as well as quaternary ammonium compounds, are ubiquitous antimicrobial agents in healthcare. Due to the highly cationic and polymeric nature of these compounds and the complex matrices in which they are found, the analytical characterization of products containing them remains challenging. In this work an efficient, sensitive, and high-resolution separation protocol was developed to perform quantitative measurements (sub-mg L-1) of alexidine dihydrochloride (ADH) and polyhexamethylene biguanide (PHMB) in commercial multipurpose contact lens solutions (MPS). Initially, contactless conductivity (C4D) detection was explored, but lacked adequate selectivity and sensitivity to quantify PHMB or ADH in commercial MPS. To overcome these limitations, an alternative approach using solid phase extraction (SPE) followed by separation with reversed phase ultra-performance liquid chromatography (RP-UPLC) was developed for both ADH and PHMB separation and detection. The most sensitive and reliable method investigated utilized standard additions to compensate for matrix effects. For ADH, concentration values measured with the presented method were consistent with data provided by the MPS manufacturer (1.6 mg L-1) within 0.10 mg L-1. PHMB quantification in MPS products was successful at concentrations <1 mg L-1 with quantitative reproducibility better than 2% RSD. Comparison of blind sample testing using the RP-UPLC method showed strong correlation (R2 = 0.939) of PHMB concentrations with results obtained by the United States Food and Drug Administration using a published HPLC-Evaporative light scattering detection (ELSD) assay. A significant advantage of this method is the ability to partially resolve PHMB polydispersity, which to date has been minimally studied and explained. By coupling with electrospray mass spectrometry (MS), a general trend was observed for increased retention as a function of PHMB chain length. The improved robustness and reproducibility of UV detection versus ELSD coupled with the superior resolving power of UPLC is an asset to the detection and characterization of PHMB and ADH. In addition to quality control of MPS, this method has potential application to the analyses skin wipes, wound dressings and other medical products where understanding how manufacturing processes lead to differences in polydispersity is important to maximize the antimicrobial properties while minimizing toxicologic effects.

Limitations in converting waste gases to fuels and chemicals.

Carbon dioxide remediation is of vital importance in mitigating the impact of greenhouse gases on climate change. While various technologies have been presented in the literature, we argue that only by valorizing CO2 capture can such technologies reach widespread adoption in the current geo-political disposition. One such option is CO2 fixation by autotrophic bacteria into bio-diesel and commodity chemicals. While proof of concept technologies have been published in the literature, yet key limitations exist, including maximal yield of aerobic CO2 fixation, and growth rates, productivities, and titers of anaerobic CO2 fixation. Researchers are currently addressing these issues through metabolic engineering and the controlled supplementation of secondary metabolites.

Characterization of recombinant OspA in two different Borrelia vaccines with respect to immunological response and its relationship to functional parameters.

BACKGROUND: Prevention of Lyme disease in dogs in North America depends on effective vaccination against infection by the tick vector-born spirochete Borrelia burgdorferi. Most vaccines effectively prevent spirochete transmission to dogs during tick feeding based on immunization with the outer-surface lipoprotein A (OspA) of B. burgdorferi. More recently, vaccines containing additional OspC protein moieties have been introduced. These are designed to enhance protection by forming a second line of defense within the vertebrate host, where OspC expression replaces OspA as the dominant surface antigen. However, supportive data for demonstration of OspC mediated protection is still lacking. Since OspA immunogenicity is of paramount importance to protection against spirochete transmission; this study was designed to compare the immunogenicity of two commercially available vaccines against the Borrelia burgdorferi OspA. We further characterized OspA antigen fractions of these vaccines with respect to their biochemical and biophysical properties. RESULTS: Two groups of beagle dogs (n = 9) were administered either: (1) a nonadjuvanted/monovalent, recombinant OspA vaccine (Recombitek® Lyme) or (2) an adjuvanted, recombinant OspA /OspC chimeric fusion vaccine (Vanguard® crLyme). The onset of the anti-OspA antibody response elicited by the nonadjuvanted/monovalent OspA vaccine was significantly earlier than that for the bivalent OspA /OspC vaccine and serum borreliacidal activity was significantly greater at all post-vaccination time points. As expected, only dogs inoculated with the bivalent OspA/OspC vaccine mounted a humoral anti-OspC response. However, only three out of nine dogs in that group had a positive response. Comparison of the OspA vaccine structures revealed that the OspA in the nonadjuvanted/monovalent vaccine was primarily in the lipidated form, eluting (SEC-HPLC) at a high molecular weight, suggestive of micelle formation. Conversely, the OspA moiety of the OspA/OspC vaccine was found to be nonlipidated and eluted as the monomeric protein. CONCLUSIONS: We hypothesize that these structural differences may account for the superior immunogenicity of the nonadjuvanted monovalent recombinant OspA vaccine in dogs over the adjuvanted OspA fraction of the OspA/OspC vaccine.

Comparative onset of immunity of oral and intranasal vaccines against challenge with Bordetella bronchiseptica.

Three groups of approximately eight-week-old beagles were vaccinated once with 1 ml of placebo vaccine (oral, n=9), 1 ml of Recombitek® Oral Bordetella (oral, n=10) or 1 ml Nobivac® Intra-Trac3 (intranasal, 0.5 ml/nostril, n=10). Seven days after vaccination, the three groups were challenged with virulent Bordetella bronchiseptica via aerosolisation. Eight of nine dogs in the placebo group and no dogs in the Recombitek® Oral Bordetella or Nobivac® Intra-Trac3 vaccine groups developed spontaneous cough of two or more consecutive days (disease case definition). Dogs in the Recombitek® Oral Bordetella and Nobivac® Intra-Trac3 groups had a significantly lower incidence of disease (P<0.0001) with a 100 per cent preventable fraction. The study demonstrated that vaccination with either Recombitek® Oral Bordetella or Nobivac® Intra-Trac3 is effective in preventing disease seven days after vaccination when compared with dogs vaccinated with a placebo.

Bone marrow mesenchymal stem cells do not enhance intra-synovial tendon healing despite engraftment and homing to niches within the synovium.

BACKGROUND: Intra-synovial tendon injuries display poor healing, which often results in reduced functionality and pain. A lack of effective therapeutic options has led to experimental approaches to augment natural tendon repair with autologous mesenchymal stem cells (MSCs) although the effects of the intra-synovial environment on the distribution, engraftment and functionality of implanted MSCs is not known. This study utilised a novel sheep model which, although in an anatomically different location, more accurately mimics the mechanical and synovial environment of the human rotator cuff, to determine the effects of intra-synovial implantation of MSCs. METHODS: A lesion was made in the lateral border of the lateral branch of the ovine deep digital flexor tendon within the digital sheath and 2 weeks later 5 million autologous bone marrow MSCs were injected under ultrasound guidance into the digital sheath. Tendons were recovered post mortem at 1 day, and 1-2, 4, 12 and 24 weeks after MSC injection. For the 1-day and 1-2-week groups, MSCs labelled with fluorescent-conjugated magnetic iron-oxide nanoparticles (MIONs) were tracked with MRI, histology and flow cytometry. The 4, 12 and 24-week groups were implanted with non-labelled cells and compared with saline-injected controls for healing. RESULTS: The MSCs displayed no reduced viability in vitro to an uptake of 20.0 ± 4.6 pg MIONs per cell, which was detectable by MRI at minimal density of ~ 3 × 104 cells. Treated limbs indicated cellular distribution throughout the tendon synovial sheath but restricted to the synovial tissues, with no MSCs detected in the tendon or surgical lesion. The lesion was associated with negligible morbidity with minimal inflammation post surgery. Evaluation of both treated and control lesions showed no evidence of healing of the lesion at 4, 12 and 24 weeks on gross and histological examination. CONCLUSIONS: Unlike other laboratory animal models of tendon injury, this novel model mimics the failed tendon healing seen clinically intra-synovially. Importantly, however, implanted stem cells exhibited homing to synovium niches where they survived for at least 14 days. This phenomenon could be utilised in the development of novel physical or biological approaches to enhance localisation of cells in augmenting intra-synovial tendon repair.

Evaluation of canine hepatic masses by use of triphasic computed tomography and B-mode, color flow, power, and pulsed-wave Doppler ultrasonography and correlation with histopathologic classification.

OBJECTIVE To determine clinical relevance for quantitative and qualitative features of canine hepatic masses evaluated by use of triphasic CT and B-mode, color flow, power, and pulsed-wave Doppler ultrasonography and to compare diagnostic accuracy of these modalities for predicting mass type on the basis of histopathologic classification. ANIMALS 44 client-owned dogs. PROCEDURES Dogs with histopathologic confirmation (needle core, punch, or excisional biopsy) of a hepatic mass were enrolled. Triphasic CT and B-mode, color flow, power, and pulsed-wave Doppler ultrasonography of each hepatic mass were performed. Seventy quantitative and qualitative variables of each hepatic mass were recorded by 5 separate observers and statistically evaluated with discriminant and stepwise analyses. Significant variables were entered in equation-based predictions for the histopathologic diagnosis. RESULTS An equation that included the lowest delayed-phase absolute enhancement of the mass and the highest venous-phase mass conspicuity was used to correctly classify 43 of 46 (93.5%) hepatic masses as benign or malignant. An equation that included only the lowest delayed-phase absolute enhancement of the mass could be used to correctly classify 42 of 46 (91.3%) masses (with expectation of malignancy if this value was < 37 Hounsfield units). For ultrasonography, categorization of the masses with cavitations as malignant achieved a diagnostic accuracy of 80.4%. CONCLUSIONS AND CLINICAL RELEVANCE Triphasic CT had a higher accuracy than ultrasonography for use in predicting hepatic lesion classification. The lowest delayed-phase absolute enhancement of the mass was a simple calculation that required 2 measurements and aided in the differentiation of benign versus malignant hepatic masses.

Pharmacological inhibition of myostatin improves skeletal muscle mass and function in a mouse model of stroke.

In stroke patients, loss of skeletal muscle mass leads to prolonged weakness and less efficient rehabilitation. We previously showed that expression of myostatin, a master negative regulator of skeletal muscle mass, was strongly increased in skeletal muscle in a mouse model of stroke. We therefore tested the hypothesis that myostatin inhibition would improve recovery of skeletal muscle mass and function after cerebral ischemia. Cerebral ischemia (45 minutes) was induced by intraluminal right middle cerebral artery occlusion (MCAO). Swiss male mice were randomly assigned to Sham-operated mice (n = 10), MCAO mice receiving the vehicle (n = 15) and MCAO mice receiving an anti-myostatin PINTA745 (n = 12; subcutaneous injection of 7.5 mg.kg-1 PINTA745 immediately after surgery, 3, 7 and 10 days after MCAO). PINTA745 reduced body weight loss and improved body weight recovery after cerebral ischemia, as well as muscle strength and motor function. PINTA745 also increased muscle weight recovery 15 days after cerebral ischemia. Mechanistically, the better recovery of skeletal muscle mass in PINTA745-MCAO mice involved an increased expression of genes encoding myofibrillar proteins. Therefore, an anti-myostatin strategy can improve skeletal muscle recovery after cerebral ischemia and may thus represent an interesting strategy to combat skeletal muscle loss and weakness in stroke patients.

In vivo visualization using MRI T2 mapping of induced osteochondrosis and osteochondritis dissecans lesions in goats undergoing controlled exercise.

In vivo visualization of subclinical osteochondrosis (OC) lesions, characterized by necrosis of epiphyseal growth cartilage, is necessary to clarify the pathogenesis of this disease. Hence, our objectives were to demonstrate induced necrosis of the epiphyseal cartilage in vivo using MRI and to monitor progression or resolution of resulting lesions. We also aimed to improve the goat model of OC by introducing controlled exercise. Vascular supply to the epiphyseal cartilage was surgically interrupted in four 5-day-old goats to induce ischemic cartilage necrosis in a medial femoral condyle. Starting 3 weeks postoperatively, goats underwent daily controlled exercise until euthanasia at 6, 10, 11 (n = 2) weeks postoperatively. T2 maps of operated and control femora were obtained in vivo at 3 (n = 4), 6 (n = 4), 9 (n = 3), and 11 (n = 2) weeks postoperatively using a 3 T MR scanner. In vivo MRI findings were validated against MRI results obtained ex vivo at 9.4 T in three goats and compared to histological results in all goats. Surgical interruption of the vascular supply caused ischemic cartilage necrosis in three out of four goats. T2 maps obtained in vivo at 3 T identified regions of increased relaxation time consistent with discrete areas of cartilage necrosis 3-11 weeks postoperatively and demonstrated delayed progression of the ossification front at 9 (n = 1) and 11 (n = 2) weeks postoperatively. In vivo MRI findings were confirmed by ex vivo MRI at 9.4 T and by histology. Identification of cartilage necrosis in clinical patients in the early stages of OC using T2 maps may provide valuable insight into the pathogenesis of this condition. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:868-875, 2017.

Ten-atom silver cluster signaling and tempering DNA hybridization.

Silver clusters with ∼10 atoms are molecules, and specific species develop within DNA strands. These molecular metals have sparsely organized electronic states with distinctive visible and near-infrared spectra that vary with cluster size, oxidation, and shape. These small molecules also act as DNA adducts and coordinate with their DNA hosts. We investigated these characteristics using a specific cluster-DNA conjugate with the goal of developing a sensitive and selective biosensor. The silver cluster has a single violet absorption band (λ(max) = 400 nm), and its single-stranded DNA host has two domains that stabilize this cluster and hybridize with target oligonucleotides. These target analytes transform the weakly emissive violet cluster to a new chromophore with blue-green absorption (λ(max) = 490 nm) and strong green emission (λ(max) = 550 nm). Our studies consider the synthesis, cluster size, and DNA structure of the precursor violet cluster-DNA complex. This species preferentially forms with relatively low amounts of Ag(+), high concentrations of the oxidizing agent O2, and DNA strands with ≳20 nucleotides. The resulting aqueous and gaseous forms of this chromophore have 10 silvers that coalesce into a single cluster. This molecule is not only a chromophore but also an adduct that coordinates multiple nucleobases. Large-scale DNA conformational changes are manifested in a 20% smaller hydrodynamic radius and disrupted nucleobase stacking. Multidentate coordination also stabilizes the single-stranded DNA and thereby inhibits hybridization with target complements. These observations suggest that the silver cluster-DNA conjugate acts like a molecular beacon but is distinguished because the cluster chromophore not only sensitively signals target analytes but also stringently discriminates against analogous competing analytes.

Safety and serological response to a matrix gene-deleted rabies virus-based vaccine vector in dogs.

Dogs account for the majority of human exposures and deaths due to rabies virus (RABV) worldwide. In this report, we show that a replication-deficient RABV-based vaccine in which the matrix gene is deleted (RABV-ΔM) is safe and induces rapid and potent VNA titers after a single inoculation in dogs. Average VNA titers peaked at 3.02 or 5.11 international units (IU/ml) by 14 days post-immunization with a single dose of 10(6) or 10(7) focus forming units (ffu), respectively, of RABV-ΔM. By day 70 post immunization, all dogs immunized with either dose of vaccine showed VNA titers >0.5IU/ml, the level indicative of a satisfactory immunization. Importantly, no systemic or local reactions were noted in any dog immunized with RABV-ΔM. The elimination of dog rabies through mass vaccination is hindered by limited resources, requirement for repeat vaccinations often for the life of a dog, and in some parts of the world, inferior vaccine quality. Our preliminary safety and immunogenicity data in dogs suggest that RABV-ΔM might complement currently used inactivated RABV-based vaccines in vaccination campaigns by helping to obtain 100% response in vaccinated dogs, thereby increasing overall vaccination coverage.

Does measurement of small intestinal diameter increase diagnostic accuracy of radiography in dogs with suspected intestinal obstruction?

The ratio between maximal small intestinal (SI) diameter and the height of the body of the fifth lumbar vertebra (L5) in radiographs has been reported as a diagnostic test in dogs with suspected intestinal obstruction. In order to assess the effect of the SI/L5 ratio on the accuracy of radiographic diagnosis of intestinal obstruction, lateral abdominal radiographs of 37 dogs with small intestinal obstruction and 48 nonobstructed dogs were mixed and examined independently by six observers who were unaware of the final diagnosis and who represented a range of experience. Observers first examined radiographs subjectively and stated the likelihood of obstruction (definitely not, probably not, equivocal, probably, definitely). Observers subsequently reexamined the radiographs, determined the SI/L5 ratio, and again stated the likelihood of obstruction. The most frequent cause of obstruction was foreign body (29/37, 78%). Dogs with SI obstruction had a significantly larger median SI/L5 ratio than nonobstructed dogs (P = 0.0002). Using an SI/L5 ratio of 1.7 for diagnosis of intestinal obstruction, sensitivity and specificity were 66%. Use of the SI/L5 ratio was not associated with increased accuracy of diagnosis for any observer, regardless of experience, hence this test may have no diagnostic impact.

Variation in foot conformation in lame horses with different foot lesions.

Foot conformation in the horse is commonly thought to be associated with lameness but scientific evidence is scarce although it has been shown in biomechanical studies that foot conformation does influence the forces acting on the deep digital flexor tendon (DDFT) and the navicular bone (NB). The aim of this study was to determine the relationships between foot conformation and different types of lesion within the foot in lame horses. It was hypothesised that certain conformation parameters differ significantly between different types of foot lesions. Conformation parameters were measured on magnetic resonance images in the mid-sagittal plane of 179 lame horses with lesions of their deep digital flexor tendon (DDFT), navicular bone (NB), collateral ligaments of the distal interphalangeal joints and other structures. Conformation parameters differed significantly between lesion groups. A larger sole angle was associated with combined DDFT and NB lesions, but not with NB lesions alone. A more acute angle of the DDFT round the NB was associated with DDFT and NB lesions, and a lower heel height index with DDFT injury. The larger the sole angle the smaller the likelihood of a DDFT or NB lesion with odds ratios of 0.86 and 0.90, respectively. This study shows an association between foot conformation and lesions but it does not allow the identification of conformation as causative factor since foot conformation may change as a consequence of lameness. Future studies will investigate foot-surface interaction in lame vs. sound horses, which may open a preventative and/or therapeutic window in foot lame horses.

Advanced imaging: use and misuse.

SUMMARY: The primary purpose of diagnostic imaging is to enable clinicians to make correct decisions about a patient's diagnosis and severity of its condition and, thereby, to contribute to effective and efficient treatment. Advanced imaging modalities are inherently better detectors of morphological lesions than radiography; hence, they offer the possibility of more accurate diagnosis and staging, with correspondingly better patient outcomes. However, there is a lack of evidence that better outcomes are being realised for veterinary patients. Furthermore, increased use of advanced imaging carries the possibility of increased misuse, primarily through unnecessary studies that contribute little to patient management other than increased costs. SCOPE: This article is intended to be relevant to all veterinary species. Advanced imaging of cats is not sufficiently developed as a specific discipline that a balanced review could be based purely on feline examples or references.