Characterisation of the main drivers of intra- and inter- breed variability in the plasma metabolome of dogs

Introduction

Dog breeds are a consequence of artificial selection for specific attributes. These closed genetic populations have metabolic and physiological characteristics that may be revealed by metabolomic analysis.

Objectives

To identify and characterise the drivers of metabolic differences in the fasted plasma metabolome and then determine metabolites differentiating breeds.

Methods

Fasted plasma samples were collected from dogs maintained under two environmental conditions (controlled and client-owned at home). The former (n = 33) consisted of three breeds (Labrador Retriever, Cocker Spaniel and Miniature Schnauzer) fed a single diet batch, the latter (n = 96), client-owned dogs consisted of 9 breeds (Beagle, Chihuahua, Cocker Spaniel, Dachshund, Golden Retriever, Greyhound, German Shepherd, Labrador Retriever and Maltese) consuming various diets under differing feeding regimens. Triplicate samples were taken from Beagle (n = 10) and Labrador Retriever (n = 9) over 3 months. Non-targeted metabolite fingerprinting was performed using flow infusion electrospray-ionization mass spectrometry which was coupled with multivariate data analysis. Metadata factors including age, gender, sexual status, weight, diet and breed were investigated.

Results

Breed differences were identified in the plasma metabolome of dogs housed in a controlled environment. Triplicate samples from two breeds identified intra-individual variability, yet breed separation was still observed. The main drivers of variance in dogs maintained in the home environment were associated with breed and gender. Furthermore, metabolite signals were identified that discriminated between Labrador Retriever and Cocker Spaniels in both environments.

Conclusion

Metabolite fingerprinting of plasma samples can be used to investigate breed differences in client-owned dogs, despite added variance of diet, sexual status and environment.

     

No psychological distress in sportsmen aged 45 years and older after cardiovascular screening,including cardiac CT: The Measuring Athlete’s Risk of Cardiovascular events (MARC) study

Background

Psychological distress caused by cardiovascular pre-participation screening (PPS) may be a reason not to implement a PPS program. We assessed the psychological impact of PPS, including cardiac computed tomography (CT), in 318 asymptomatic sportsmen aged ≥45 years.

Methods

Coronary artery disease (CAD) was defined as a coronary artery calcium score ≥100 Agatson units and/or ≥50% luminal stenosis on contrast-enhanced cardiac CT. Psychological impact was measured with the Impact of Event Scale (IES) (seven items) on a six-point scale (grade 0–5). A sum score ≥19 indicates clinically relevant psychological distress. A Likert scale was used to assess overall experiences and impact on sports and lifestyle.

Results

A total of 275 participants (86.5% response rate, 95% CI 83–90%) with a mean age of 54.5 ± 6.4 years completed the questionnaires, 48 (17.5%, 95% CI 13–22%) of whom had CAD. The median IES score was 1 (IQR 0–2, [0–23]). IES was slightly higher in those with CAD (mean rank 175 vs. 130, p < 0.001). One participant (with CAD) experienced clinically relevant psychological distress (IES = 23). Participants reported numerous benefits, including feeling safer exercising (58.6%, 95% CI 53–65%) and positive lifestyle changes, especially in those with CAD (17.2 vs. 52.1%, p < 0.001). The majority were satisfied with their participation (93.8%, 95% CI 91–97%).

Conclusion

Cardiovascular PPS, including cardiac CT, causes no relevant psychological distress in older sportsmen. Psychological distress should not be a reason to forego screening in sportsmen.

     

Cetirizine per os: exposure and antihistamine effect in the dog

Background

Cetirizine is an antihistamine used in dogs, but plasma concentrations in relation to effect after oral administration are not well studied. This study investigated cetirizine exposure and the plasma cetirizine concentration-antihistamine response relation in the dog following oral administration of cetirizine.

Results

Eight Beagle dogs were included in a cross-over study consisting of two treatments. In treatment one, cetirizine 2–4 mg/kg was administered per os once daily for 3 days. The other treatment served as a control. Wheal diameter induced by intra-dermal histamine injections served as response-biomarker. Cetirizine plasma concentration was quantified by UHPLC-MS/MS. Median (range) cetirizine plasma terminal half-life was 10 h (7.9–16.5). Cetirizine significantly inhibited wheal formation compared with the premedication baseline. Maximum inhibition of wheal formation after treatment with cetirizine per os was 100% compared with premedication wheal diameter. The median (range) IC₅₀-value for reduction in wheal area was 0.33 µg/mL (0.07–0.45). The median (range) value for the sigmoidicity factor was 1.8 (0.8–3.5). A behavioral study was also conducted and revealed no adverse effects, such as sedation.

Conclusion

The results indicate that a once-daily dosing regimen of 2–4 mg/kg cetirizine per os clearly provides a sufficient antihistamine effect. Based on this experimental protocol, cetirizine may be an option to treat histamine-mediated inflammation in the dog based on this experimental protocol but additional clinical studies are required.

     

Diabetes screening intervals based on risk stratification

Background

Guidelines for frequency of Type 2 diabetes mellitus (DM) screening remain unclear, with proposed screening intervals typically based on expert opinion. This study aims to demonstrate that HbA1c screening intervals may differ substantially when considering individual risk for diabetes.

Methods

This was a multi-institutional retrospective open cohort study. Data were collected between April 1999 to March 2014 from one urban and one rural cohort in Japan. After categorization by age, we stratified individuals based on cardiovascular disease risk (Framingham 10-year cardiovascular risk score) and body mass index (BMI). We adapted a signal-to-noise method for distinguishing true HbA1c change from measurement error by constructing a linear random effect model to calculate signal and noise of HbA1c. Screening interval for HbA1c was defined as informative when the signal-to-noise ratio exceeded 1.

Results

Among 96,456 healthy adults, 46,284 (48.0%) were male; age (range) and mean HbA1c (SD) were 48 (30–74) years old and 5.4 (0.4)%, respectively. As risk increased among those 30–44 years old, HbA1c screening intervals for detecting Type 2 DM consistently decreased: from 10.5 (BMI <18.5) to 2.4 (BMI?>?30) years, and from 8.0 (Framingham Risk Score <10%) to 2.0 (Framingham Risk Score ≥20%) years. This trend was consistent in other age and risk groups as well; among obese 30–44 year olds, we found substantially shorter intervals compared to other groups.

Conclusion

HbA1c screening intervals for identification of DM vary substantially by risk factors. Risk stratification should be applied when deciding an optimal HbA1c screening interval in the general population to minimize overdiagnosis and overtreatment.

     

Neonatal tooth with Riga-Fide disease affecting breastfeeding: a case report

Background

Neonatal teeth erupt during the neonatal period and natal teeth are the presence of teeth since birth. While rare, natal teeth and neonatal teeth can have a significant impact on breastfeeding. Neonatal teeth are less common, and although its exact etiology is still unknown, it can cause difficulties in breastfeeding to the mother and may eventually lead to discontinuation of breastfeeding. Other associated possible complications include tooth aspiration and sublingual ulceration. This paper was aimed to discuss the clinical features, complications, and management of neonatal tooth, in addition to its impact on breastfeeding and role in sublingual ulcer formation.

Case presentation

We present a baby girl who had a neonatal tooth with sublingual ulceration (Riga-Fede disease), which resulted in a difficulty to breastfeed for the baby and nipple pain to the mother. Following the extraction of the baby’s tooth, she immediately continued breastfeeding, and her tongue ulcer healed well.

Conclusion

Extraction of the neonatal tooth promoted rapid healing of oral ulcers and the reestablishment of breastfeeding.

     

Markov modeling in hepatitis B screening and linkage to care

Background

With up to 240 million people chronically infected with hepatitis B worldwide, including an estimated 2 million in the United States, widespread screening is needed to link the infected to care and decrease the possible consequences of untreated infection, including liver cancer, cirrhosis and death. Screening is currently fraught with challenges in both the developed and developing world. New point-of-care tests may have advantages over standard-of-care tests in terms of cost-effectiveness and linkage to care. Stochastic modeling is applied here for relative utility assessment of point-of-care tests and standard-of-care tests for screening.

Methods

We analyzed effects of point-of-care versus standard-of-care testing using Markov models for disease progression in individual patients. Simulations of large cohorts with distinctly quantified models permitted the assessment of particular screening schemes. The validity of the trends observed is supported by sensitivity analyses for the simulation parameters.

Results

Increased utilization of point-of-care screening was shown to decrease hepatitis B-related mortalities and increase life expectancy at low projected expense.

Conclusions

The results suggest that standard-of-care screening should be substituted by point-of-care tests resulting in improved linkage to care and decrease in long-term complications.

     

Urinary Metabolomic Signatures in Obstructive Sleep Apnea through Targeted Metabolomic Analysis: A Pilot Study

Introduction

Obstructive sleep apnea (OSA) is very common sleep problem, and it is associated with serious morbidities such as cardiovascular diseases and metabolic diseases. Overnight polysomnography (PSG) is the gold standard test for OSA, but it is expensive and requires specific facilities and equipment. Thus, novel screening methods are needed for effective diagnosis and follow-up in OSA.

Objectives

The aims of the study were to investigate the urinary metabolic signatures and identify potential urine markers for OSA using a mass spectrometry (MS)-based assay for targeted metabolomics.

Methods

Urine samples were collected from 48 male subjects who visited a sleep clinic for suspicious OSA. All underwent overnight in-laboratory polysomnography. The Biocrates AbsoluteIDQ p180 kit was used for targeted metabolomics.

Results

Among the 86 metabolites quantified, three acylcarnitines, one biogenic amine, two glycerophospholipids, and two sphingomyelins were differently expressed in OSA patients [apnea-hypopnea index (AHI) ≥5] compared with control groups (AHI <5 and/or simple snoring with no other sleep disorders). Additional partial correlation and multivariate logistic regression analysis revealed that long-chain acylcarnitine C14:1, symmetric dimethylarginine, and sphingomyelin C18:1 might be potential biomarkers for OSA. Receiver operating characteristic analysis showed favorable predictive properties of these metabolites. Furthermore, a combination of the metabolites exceeding cutoff values yielded further improved sensitivity or specificity.

Conclusions

MS-based targeted metabolomics identified specific classes of urinary metabolites that were up-regulated in OSA patients. Further assessments in large populations are required to clarify the screening values of these metabolite markers.

     

Identification of trunk mutations in gastric carcinoma: a case study

Background

Intratumor heterogeneity (ITH) poses an urgent challenge for cancer precision medicine because it can cause drug resistance against cancer target therapy and immunotherapy. The search for trunk mutations that are present in all cancer cells is therefore critical for each patient.

Case presentation

In this study, we aimed to evaluate the efficiency of multiregional sequencing for the identification of trunk mutations present in all regions of a tumor as a case study. We applied multiregional whole-exome sequencing (WES) to investigate the genetic heterogeneity and homogeneity of a case of gastric carcinoma. Approximately 83% of common missense mutations present in two samples and approximately 89% of common missense mutations present in three samples were trunk mutations. Notably, trunk mutations appeared to have higher variant allele frequencies (VAFs) than non-trunk mutations.

Conclusions

Our results indicate that small-scale multiregional sampling and subsequent screening of low VAF somatic mutations might be a cost-effective strategy for identifying the majority of trunk mutations in gastric carcinoma.

     

Kinetic gait analysis in English Bulldogs

Background

Canine breed conformation may interfere with locomotion and may predispose to orthopedic disease. Bulldogs have a high incidence of orthopedic diseases such as hip dysplasia. Kinetic gait analysis provides an objective way to assess and analyze locomotion. The aim of this study was to study the vertical forces of English Bulldogs during walk using a pressure sensitive walkway. We hypothesize that Bulldogs affected by orthopedic diseases have decreased weight bearing and asymmetric locomotion in the limbs despite having mild to no sings during clinical examination. Thirty English Bulldogs were tested. Peak vertical force, vertical impulse, rate of loading, stance phase duration, symmetry index, goniometry and incidence of orthopedic diseases were recorded.

Results

Although none of the dogs showed signs of pain or discomfort upon manipulation of the hip joints, all dogs had radiographic evidences of hip dysplasia and lack of significant peak vertical force, vertical impulse and stance time differences. The dogs had a mean hind limb symmetry index of 19.8 ± 19.5% and rates of loading ranged from 1.0 to 3.1.

Conclusions

Despite the lack of evident decrease in weight bearing, subclinical lameness can be inferred. The examined dogs had a mean hind limb symmetry index of 19.8 ± 19.5%. Symmetry indices reported in dogs free from orthopedic diseases range from 0.3 to 9.6%. Given non-lame dogs are expected to have a symmetry index close to 0%, data from this study suggests that Bulldogs have gait dysfunctions, which translates into hind limb asymmetries and rate of loading was consistent with severe hip dysplasia despite no visible signs of gait dysfunction. Future studies comparing lame and non-lame Bulldogs are warranted.

     

Prospects for developing an accurate diagnostic biomarker panel for low prevalence cancers

Background

Early detection screening of asymptomatic populations for low prevalence cancers requires a highly specific test in order to limit the cost and anxiety produced by falsely positive identifications. Most solid cancers are a heterogeneous collection of diseases as they develop from various combinations of genetic lesions and epigenetic modifications. Therefore, it is unlikely that a single test will discriminate all cases of any particular cancer type. We propose a novel, intuitive biomarker panel design that accommodates disease heterogeneity by allowing for diverse biomarker selection that increases diagnostic accuracy.

Methods

Using characteristics of nine pancreatic ductal adenocarcinoma (PDAC) biomarkers measured in human sera, we modeled the behavior of biomarker panels consisting of a sum of indicator variables representing a subset of biomarkers within a larger biomarker data set. We then chose a cutoff for the sum to force specificity to be high and delineated the number of biomarkers required for adequate sensitivity of PDAC in our panel design.

Results

The model shows that a panel consisting of 40 non-correlated biomarkers characterized individually by 32% sensitivity at 95% specificity would require any 7 biomarkers to be above their respective thresholds and would result in a panel specificity and sensitivity of 99% each.

Conclusions

A highly accurate blood-based diagnostic panel can be developed from a reasonable number of individual serum biomarkers that are relatively weak classifiers when used singly. A panel constructed as described is advantageous in that a high level of specificity can be forced, accomplishing a prerequisite for screening asymptomatic populations for low-prevalence cancers.

     

Identification of canine norovirus in dogs in South Korea

Background

Canine noroviruses (CaNoVs) are classified into genogroups GIV, GVI, and GVII and have been detected in fecal samples from dogs since their first appearance in a dog with enteritis in Italy in 2007. CaNoVs may be a public health concern because pet animals are an integral part of the family and could be a potential reservoir of zoonotic agents. Nonetheless, there was no previous information concerning the epidemiology of CaNoV in South Korea. In the present study, we aimed to detect CaNoV antigens and to investigate serological response against CaNoV in dogs.

Results

In total, 459 fecal samples and 427 sera were collected from small animal clinics and animal shelters housing free-roaming dogs in geographically distinct areas in South Korea. For the detection of CaNoV, RT-PCR was performed using target specific primers, and nucleotide sequences of CaNoV isolates were phylogenetically analyzed. Seroprevalence was performed by ELISA based on P domain protein. CaNoVs were detected in dog fecal samples (14/459, 3.1%) and were phylogenetically classified into the same cluster as previously reported genogroup GIV CaNoVs. Seroprevalence was performed, and 68 (15.9%) of 427 total dog serum samples tested positive for CaNoV IgG antibodies.

Conclusion

This is the first study identifying CaNoV in the South Korean dog population.

     

Comparison of two prepill cortisol concentrations in dogs with hypercortisolism treated with trilostane

Background

The ideal method for monitoring trilostane therapy in dogs with hypercortisolism is still open to debate. Recently, determination of the pre-trilostane (prepill) cortisol concentration has been proposed to be more repeatable than either post-trilostane or post-ACTH cortisol. The aim of this study was to compare two prepill cortisol concentrations in dogs with hypercortisolism during trilostane therapy. Sixteen client-owned dogs with naturally occurring hypercortisolism were prospectively included and cortisol concentrations were measured twice, 1?h apart, before the morning trilostane dose (prepill 1 and 2 cortisol).

Results

A total of 47 prepill cortisol measurement pairs were included. Compared to prepill 1, prepill 2 cortisol was higher in 15, equal in 8 and lower in 24 pairs. Group agreement between prepill 1 and 2 cortisol was 70% (moderate agreement - weighted kappa 0.55). In 30% of the pairs, group assignment was discrepant, implying a different therapeutic decision. In some dogs certain circumstances (e.g. excessive barking, difficulties during blood collection, excitement at arrival) were identified as potential factors explaining the discrepancy between prepill 1 and 2 cortisol measurements.

Conclusions

In a substantial number of dogs treated with trilostane, the two prepill cortisol concentrations differed. Part of this difference might be ascribable to stressful events during test performance. When using prepill cortisol measurements to monitor trilostane therapy, recording of any incident during handling that might affect cortisol release might be helpful to make a reliable decision about a trilostane dose adaptation.

     

A prospective observational cohort study in primary care practices to identify factors associated with treatment failure in <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> skin and soft tissue infections

Background

The incidence of outpatient visits for skin and soft tissue infections (SSTIs) has substantially increased over the last decade. The emergence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has made the management of S. aureus SSTIs complex and challenging. The objective of this study was to identify risk factors contributing to treatment failures associated with community-associated S. aureus skin and soft tissue infections SSTIs.

Methods

This was a prospective, observational study among 14 primary care clinics within the South Texas Ambulatory Research Network. The primary outcome was treatment failure within 90 days of the initial visit. Univariate associations between the explanatory variables and treatment failure were examined. A generalized linear mixed-effect model was developed to identify independent risk factors associated with treatment failure.

Results

Overall, 21% (22/106) patients with S. aureus SSTIs experienced treatment failure. The occurrence of treatment failure was similar among patients with methicillin-resistant S. aureus and those with methicillin-susceptible S. aureus SSTIs (19 vs. 24%; p = 0.70). Independent predictors of treatment failure among cases with S. aureus SSTIs was a duration of infection of ≥7 days prior to initial visit [aOR, 6.02 (95% CI 1.74–19.61)] and a lesion diameter size ≥5 cm [5.25 (1.58–17.20)].

Conclusions

Predictors for treatment failure included a duration of infection for ≥7 days prior to the initial visit and a wound diameter of ≥5 cm. A heightened awareness of these risk factors could help direct targeted interventions in high-risk populations.

     

Investigation of urinary volatomic alterations in head and neck cancer: a non-invasive approach towards diagnosis and prognosis

Introduction

Head and neck cancer (HNC), like many other forms of cancer, is usually detected in advanced stages, causing poor survival outcomes. Lack of specific and sensitive screening markers for early detection of HNC has worsened the scenario for the patients as well as the clinicians. Therefore, identification of efficient, noninvasive and affordable screening marker/methodology with high specificity and sensitivity is imminent need of situation.

Objectives

This study aims to identify and characterize urinary volatomic alterations specific to HNC.

Methods

Volatomic analysis of urine samples collected from HNC patients (n?=?29) and healthy controls (n?=?31) was performed using headspace solid phase microextraction coupled to gas chromatography mass spectrometry (GC–MS). Both univariate and multivariate statistical approaches were used to investigate HNC specific volatomic alterations.

Results

Statistical analysis revealed a total of 28 metabolites with highest contribution towards discrimination of HNC patients from healthy controls (VIP >1, p?<?0.05, Log₂ FC ≥0.58/≤?0.57). The discrimination efficiency and accuracy of urinary VOCs was ascertained by ROC curve analysis that allowed the identification of four metabolites viz. 2,6-dimethyl-7-octen-2-ol, 1-butanol, p-xylene and 4-methyl-2-heptanone with highest sensitivity and specificity to discriminate HNC patients from healthy controls. Further, the metabolic pathway analysis identified several dysregulated pathways in HNC patients and their detailed investigations could unravel novel mechanistic insights into the disease pathophysiology.

Conclusion

Overall, this study provides valuable fingerprint of the volatile profile of HNC patients, which in turn, might help in improving the current understanding of this form of cancer and lead to the development of non-invasive approaches for HNC diagnosis.

     

Interactions between cognitive and sensory load while planning and controlling complex gait adaptations in Parkinson’s disease

Background

Recent research has argued that removal of relevant sensory information during the planning and control of simple, self-paced walking can result in increased demand on central processing resources in Parkinson’s disease (PD). However, little is known about more complex gait tasks that require planning of gait adaptations to cross over an obstacle in PD.

Methods

In order to understand the interaction between availability of visual information relevant for self-motion and cognitive load, the current study evaluated PD participants and healthy controls while walking toward and stepping over an obstacle in three visual feedback conditions: (i) no visual restrictions; (ii) vision of the obstacle and their lower limbs while in complete darkness; (iii) vision of the obstacle only while in complete darkness; as well as two conditions including a cognitive load (with a dual task versus without a dual task). Each walk trial was divided into an early and late phase to examine changes associated with planning of step adjustments when approaching the obstacle.

Results

Interactions between visual feedback and dual task conditions during the obstacle approach were not significant. Patients with PD had greater deceleration and step time variability in the late phase of the obstacle approach phase while walking in both dark conditions compared to control participants. Additionally, participants with PD had a greater number of obstacle contacts when vision of their lower limbs was not available specifically during the dual task condition. Dual task performance was worse in PD compared to healthy control participants, but notably only while walking in the dark regardless of visual feedback.

Conclusions

These results suggest that reducing visual feedback while approaching an obstacle shifts processing to somatosensory feedback to guide movement which imposes a greater demand on planning resources. These results are key to fully understanding why trips and falls occur in those with PD.

     

Quantification and recognition of parkinsonian gait from monocular video imaging using kernel-based principal component analysis

Background

The computer-aided identification of specific gait patterns is an important issue in the assessment of Parkinson's disease (PD). In this study, a computer vision-based gait analysis approach is developed to assist the clinical assessments of PD with kernel-based principal component analysis (KPCA).

Method

Twelve PD patients and twelve healthy adults with no neurological history or motor disorders within the past six months were recruited and separated according to their "Non-PD", "Drug-On", and "Drug-Off" states. The participants were asked to wear light-colored clothing and perform three walking trials through a corridor decorated with a navy curtain at their natural pace. The participants' gait performance during the steady-state walking period was captured by a digital camera for gait analysis. The collected walking image frames were then transformed into binary silhouettes for noise reduction and compression. Using the developed KPCA-based method, the features within the binary silhouettes can be extracted to quantitatively determine the gait cycle time, stride length, walking velocity, and cadence.

Results and Discussion

The KPCA-based method uses a feature-extraction approach, which was verified to be more effective than traditional image area and principal component analysis (PCA) approaches in classifying "Non-PD" controls and "Drug-Off/On" PD patients. Encouragingly, this method has a high accuracy rate, 80.51%, for recognizing different gaits. Quantitative gait parameters are obtained, and the power spectrums of the patients' gaits are analyzed. We show that that the slow and irregular actions of PD patients during walking tend to transfer some of the power from the main lobe frequency to a lower frequency band. Our results indicate the feasibility of using gait performance to evaluate the motor function of patients with PD.

Conclusion

This KPCA-based method requires only a digital camera and a decorated corridor setup. The ease of use and installation of the current method provides clinicians and researchers a low cost solution to monitor the progression of and the treatment to PD. In summary, the proposed method provides an alternative to perform gait analysis for patients with PD.

     

The Mini-BESTest - a clinically reproducible tool for balance evaluations in mild to moderate Parkinson’s disease?

Background

The Mini-BESTest is a clinical balance test that has shown a high sensitivity in detecting balance impairments in elderly with Parkinson's disease (PD). However, its reproducibility between different raters and between test occasions has yet to be investigated in a clinical context. Moreover, no one has investigated the reproducibility of the Mini-BESTest's subcomponents (i.e. anticipatory postural adjustments; postural responses; sensory orientation and dynamic gait).We aimed to investigate the inter-rater and test-retest reproducibility (reliability as well as agreement) of the Mini-BESTest, as well as its subcomponents, in elderly with mild to moderate PD, performed under conditions assimilating clinical practice.

Method

This was an observational measurement study with a test-retest design.Twenty-seven individuals with idiopathic PD (66 - 80 years, mean age: 73; Hoehn & Yahr: 2-3; 1-15 years since diagnosis) were included. Two test administrators, having different experiences with the Mini-BESTest, administered the test individually, in separate rooms in a hospital setting. For the test-retest assessment, all participants returned 7 days after the first test session to perform the Mini-BESTest under similar conditions. Intra-class correlation coefficients (ICC_2.1), standard error of measurement (SEM_agreement), and smallest real difference (SRD) were analyzed.

Results

The Mini-BESTest showed good reliability for both inter-rater and test-retest reproducibility (ICC = 0.72 and 0.80). Regarding agreement, the measurement error (SRD) was found to be 4.1 points (accounting for 15% of the maximal total score) for inter-rater reproducibility and 3.4 points (12% of the maximal total score) for test-retest reproducibility. The investigation of the Mini-BESTest's subcomponents showed a similar pattern for both inter-rater and test-retest reproducibility, where postural responses had the largest proportional measurement error, and sensory orientation showed the highest agreement.

Conclusions

Our findings indicate that the Mini-BESTest is able to distinguish between individuals with mild to moderate PD; however, when used in clinical balance assessments, the large measurement error needs to be accounted for.

     

Boosting fatty acid synthesis in <Emphasis Type="Italic">Rhodococcus opacus</Emphasis> PD630 by overexpression of autologous thioesterases

Objectives

To explore the role of thioesterases in Rhodococcus opacus PD630 by endogenously overexpression in this bacteria for increased lipid production.

Results

Overexpression of four thioesterases from R. opacus PD630 in E. coli led to a 2- to 8-fold increase in C16:1 and C18:1 fatty acids while, when overexpressed in R. opacus PD630, only two recombinants had significant effect on the quantities and compositions of total fatty acid. The contents of total fatty acids (FAs) in two recombinants, pJTE2 (OPAG_00508 thioesterase) and pJTE4 (WP_012687673.1 thioesterase), were 400–460 mg/g (CDW) which is 1.5 times of wild-type strain PD630 (300-350 mg/g CDW), and 20–30 % (w/w) more than that of the control strain PDpJAM2 (330-370 mg/g CDW). The contents of 17:1 and 18:1 fatty acids increased by about 27 and 35 %, respectively, in pJTE2 and by 35 and 20 %, respectively, in pJTE4 compared with the control strain.

Conclusions

The engineered strains showed improved production of lipid (as total fatty acids), and could also tailor the composition of the fatty acid profile when cultured in mineral salts medium using glucose as sole carbon source.

     

Head and pelvic vertical displacement in dogs with induced swinging limb lameness: an experimental study

Background

Swinging limb lameness is defined as a motion disturbance ascribed to a limb in swing phase. Little is known about its biomechanics in dogs, particularly about the body motions that accompany it, such as vertical head and pelvic motion asymmetry. The aim of this study was to describe the changes in vertical head and pelvic motion asymmetry in dogs with induced swinging limb motion disturbance, mimicking a swinging limb lameness. Fore- and hind-limb lameness was induced in ten sound dogs by placing a weight (200 g) proximal to the carpus or tarsus, respectively. Marker-based motion capture by eight infrared light emitting video cameras recorded the dogs when trotting on a treadmill. Body symmetry parameters were calculated, including differences between the two highest positions of the head (HDmax) and pelvis (PDmax) and between the two lowest positions of the head (HDmin) and pelvis (PDmin), with a value of zero indicating perfect symmetry.

Results

Induction of swinging forelimb lameness showed significant changes in HDmax (median and range: sound 1.3 mm [??4.7 to 3.1], in the left side ??28.5 mm [??61.2 to ??17.9] and in the right side 20.1 mm [??4.4 to 47.5]) and, induction of swinging hind limb lameness showed significant changes in PDmax (sound 2.7 mm [??7.4 to 7.2], in the left side ??10.9 mm [??22.4 to 0.5] and in the right side 8.6 mm [??3 to 30]), as well as an increased hip movement asymmetry (sound 1.6 mm [??8.6 to 19.9], in the left side ??18.1 mm [??36.7 to 5.4] and in the right side 15 mm [??20.7 to 32.1]) (P?<?0.05).

Conclusions

Induced swinging fore- and hind limb lameness resulted in significant increased asymmetry of the maximal vertical displacement movement of the head and pelvis, due to decreased lifting of the head in forelimb lameness and of the pelvis in hind limb lameness. The results suggest that asymmetry of the maximal vertical displacement of the head and pelvis (i.e. lifting) is a key lameness sign to evaluate during examination of swinging limb lameness.