Multiple studies implicate heterozygous
GBA
mutations as a major genetic risk factor for Parkinson's disease (PD); however, the frequency of mutations has never been examined in PD patients from the ...Irish population. We prospectively recruited 314 unrelated Irish PD patients (UK Brain Bank Criteria) and 96 Irish healthy controls (without any signs or family history of parkinsonism) attending. The Dublin Neurological Institute (DNI). Complete exon
GBA
Sanger sequencing analysis with flanking intronic regions was performed. The
GBA
carrier frequency was 8.3% in PD and 3.1% in controls. We identified a number of potentially pathogenic mutations including a p.G195E substitution and a p.G377C variant, previously described in a case study of Gaucher's disease in Ireland. On genotype–phenotype assessment hallucinations, dyskinesia, and dystonia were more prevalent in
GBA
-PD. The genetic etiology of PD in Ireland differs from the continental Europe as seen with the lower
LRRK2
and higher than in most European countries
GBA
mutation frequency. Determining genetic risk factors in different ethnicities will be critical for future personalized therapeutic approach.
The objectives of this study were to evaluate efficacy of a 2-dose regimen of ceftiofur crystalline free acid sterile suspension (CCFA-SS) for treatment of acute metritis in lactating dairy cows ...under field conditions and to provide additional safety and injection site tolerance data for injections at the base of the ear. Cows at 15 dairies with rectal temperature ≥39.5°C and fetid uterine discharge ≤10 d postcalving were randomly assigned by blocks of 2, based on order of entry and without regard to parity, to treatment with saline (1.5mL/45.5kg of body weight, n=509) or CCFA-SS (6.6mg of ceftiofur equivalents/kg of body weight, n=514). Treatments were administered by subcutaneous injection in the posterior aspect of the ear where it attaches to the head; the first dose was administered on study d 0 and the second dose was administered in the contra lateral ear on study d 3. Rectal temperatures were recorded on study d 1 to 4 and 5 or 6 and cows were clinically evaluated daily from study d 1 to 13. Cows that exhibited increased adverse clinical signs of poor health or complications associated with metritis were categorized as a treatment failure and administered escape therapy. Each cow received a veterinary physical examination on study d 5 or 6 to determine if she should be removed from the study and on study d 14 to determine clinical cure or failure to cure. Clinical cure was defined as rectal temperature <39.5°C and non-fetid and purulent or mucopurulent discharge on study d 14 and no escape therapy administered. The injection procedure was scored after each injection (study d 0 and 3) and injection sites and ear carriage were scored on study d 5 or 6, 14, and 57±3. Of the 1,023 cows enrolled, 7 were completely censored due to protocol deviations and 34 were removed for protocol deviations or medical conditions not related to metritis. Clinical cure rate was higher for CCFA-SS than for saline (74.3 vs. 55.3%) and rectal temperatures for each of study d 1 to 5 or 6 were lower for CCFA-SS than saline. Injection procedure indices showed that CCFA-SS could be practically and safely administered using commercial dairy facilities. Although injection site scores were higher for CCFA-SS than saline at study d 5 or 6 and 14, ≥98.6% of ears were normal on d 57±3. Thus, a 2-dose treatment with CCFA-SS given 72h apart increased metritis clinical cure rate and was well tolerated in dairy cows.
Ephemeral wetlands are globally important systems that are regulated by regular cycles of wetting and drying, which are primarily controlled by responses to relatively short-term weather events (
...e.g.
, precipitation and evapotranspiration). Climate change is predicted to have significant effects on many ephemeral wetland systems and the organisms that depend on them through altered filling or drying dates that impact hydroperiod. To examine the potential effects of climate change on pine flatwoods wetlands in the southeastern United States, we created statistical models describing wetland hydrologic regime using an approximately 8-year history of water level monitoring and a variety of climate data inputs. We then assessed how hydrology may change in the future by projecting models forward (2025–2100) under six future climate scenarios (three climate models each with two emission scenarios). We used the model results to assess future breeding conditions for the imperiled Reticulated Flatwoods Salamander (
Ambystoma bishopi
), which breeds in many of the study wetlands. We found that models generally fit the data well and had good predictability across both training and testing data. Across all models and climate scenarios, there was substantial variation in the predicted suitability for flatwoods salamander reproduction. However, wetlands with longer hydroperiods tended to have fewer model iterations that predicted at least five consecutive years of reproductive failure (an important metric for population persistence). Understanding potential future risk to flatwoods salamander populations can be used to guide conservation and management actions for this imperiled species.
Understanding natural variation in stream phosphorus (P) concentrations over space and time is critical for understanding natural drivers of catchment behavior and establishing regulatory standards. ...Across minimally impacted benchmark streams (n = 81) in Florida, spatial variation in mean total P concentrations was large, indicating the importance of geologic controls on catchment solute dynamics. While this variation was significantly predicted by geographic regions, within regions we observed nearly comparable cross‐site variation, suggesting important finer‐scale heterogeneity in baseline catchment chemistry. Within‐site residual variation (unexplained by region or site) was as large as spatial variation, suggesting temporal variation in response to drivers such as flow may be critically important. To further explore timescales of P export variation, we collected long‐term, high‐frequency (subdaily) measurements of stream discharge (Q) and soluble reactive P (SRP) in 2 forested watersheds. We observed significant variation at annual, event, and diel timescales, all of which arise primarily from corresponding Q‐variation. Over the entire period of record, we generally observed a strong dilution signal, with SRP concentrations declining with increased Q. Despite significant SRP variation, flow variation was far larger and, thus, dominated temporal control on downstream flux. Within‐storm events, we observed strong and consistent clockwise SRP versus Q hysteresis, suggesting mobilization of proximal SRP stores. Diel variation exhibited mid‐afternoon concentration minima, Q‐controlled amplitude, and pronounced seasonal shifts in both magnitude and timing consistent with riparian evapotranspiration‐regulating lateral inputs of P‐rich groundwater. Such high‐resolution temporal signals allow identification of solute sources and provide insights into geologic and hydrologic drivers of solute variation.
This study evaluated the effect of a six-week deep slow breathing (DSB) program on pain, physical function, and heart rate variability (HRV) in subjects with lower extremity joint pain. Twenty ...subjects were assigned into training (n = 10) and control (n = 10) groups. The training group participated in a six-week DSB program consisting of weekly training sessions and at-home breathing exercises. DSB exercises focused on prolonging the exhalation and the pause following exhalation. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) was used to assess pain and physical function, and HRV data were obtained before and after intervention. Results revealed no significant interactions between group and time for any of the variables. There was no significant main effect for group, but there was a significant main effect (p<0.025) and a large effect size for time on both pain (ηp2 = 0.454) and physical function (ηp2 = 0.506). There were no significant main effects (p>0.017) for group and time on LF power (group ηp2 = 0.039, time ηp2 = 0.061), HF power (group ηp2 = 0.039, time ηp2 = 0.039), and LF/HF ratio (group ηp2 = 0.036, time ηp2 = 0.169). Results indicated that the six-week DSB program was not sufficient to alleviate pain or improve physical function in subjects with lower extremity joint pain. Although the pain was not alleviated, other beneficial effects such as better coping with the pain were reported in the majority of training subjects. As this is the first study to examine the use of DSB for lower extremity joint pain and dysfunction, further research is needed to investigate the efficacy and applicability of DSB.
Background: While frontotemporal involvement is increasingly recognized in Amyotrophic lateral sclerosis (ALS), the degeneration of limbic networks remains poorly characterized, despite growing ...evidence of amnestic deficits, impaired emotional processing and deficits in social cognition. Methods: A prospective neuroimaging study was conducted with 204 individuals with ALS and 111 healthy controls. Patients were stratified for hexanucleotide expansion status in C9orf72. A deep-learning-based segmentation approach was implemented to segment the nucleus accumbens, hypothalamus, fornix, mammillary body, basal forebrain and septal nuclei. The cortical, subcortical and white matter components of the Papez circuit were also systematically evaluated. Results: Hexanucleotide repeat expansion carriers exhibited bilateral amygdala, hypothalamus and nucleus accumbens atrophy, and C9orf72 negative patients showed bilateral basal forebrain volume reductions compared to controls. Both patient groups showed left rostral anterior cingulate atrophy, left entorhinal cortex thinning and cingulum and fornix alterations, irrespective of the genotype. Fornix, cingulum, posterior cingulate, nucleus accumbens, amygdala and hypothalamus degeneration was more marked in C9orf72-positive ALS patients. Conclusions: Our results highlighted that mesial temporal and parasagittal subcortical degeneration is not unique to C9orf72 carriers. Our radiological findings were consistent with neuropsychological observations and highlighted the importance of comprehensive neuropsychological testing in ALS, irrespective of the underlying genotype.
Background
Breast cancer patients report a perception of increased muscle fatigue, which can persist following surgery and standardized therapies. In a clinical experiment, we tested the hypothesis ...that pathways regulating skeletal muscle fatigue are down‐regulated in skeletal muscle of breast cancer patients and that different muscle gene expression patterns exist between breast tumour subtypes. In a preclinical study, we tested the hypothesis that mammary tumour growth in mice induces skeletal muscle fatigue and that overexpression of the cytokine interleukin‐15 (IL‐15) can attenuate mammary tumour‐induced muscle fatigue.
Methods
Early stage non‐metastatic female breast cancer patients (n = 14) and female non‐cancer patients (n = 6) provided a muscle biopsy of the pectoralis major muscle during mastectomy, lumpectomy, or breast reconstruction surgeries. The breast cancer patients were diagnosed with either luminal (ER+/PR+, n = 6), triple positive (ER+/PR+/Her2/neu+, n = 5), or triple negative (ER−/PR−/Her2/neu−, n = 3) breast tumours and were being treated with curative intent either with neoadjuvant chemotherapy followed by surgery or surgery followed by standard post‐operative therapy. Biopsies were used for RNA‐sequencing to compare the skeletal muscle gene expression patterns between breast cancer patients and non‐cancer patients. The C57BL/6 mouse syngeneic mammary tumour cell line, E0771, was used to induce mammary tumours in immunocompetent mice, and isometric muscle contractile properties and fatigue properties were analysed following 4 weeks of tumour growth.
Results
RNA‐sequencing and subsequent bioinformatics analyses revealed a dysregulation of canonical pathways involved in oxidative phosphorylation, mitochondrial dysfunction, peroxisome proliferator‐activated receptor signalling and activation, and IL‐15 signalling and production. In a preclinical mouse model of breast cancer, the rate of muscle fatigue was greater in mice exposed to mammary tumour growth for 4 weeks, and this greater muscle fatigue was attenuated in transgenic mice that overexpressed the cytokine IL‐15.
Conclusions
Our data identify novel genes and pathways dysregulated in the muscles of breast cancer patients with early stage non‐metastatic disease, with particularly aberrant expression among genes that would predispose these patients to greater muscle fatigue. Furthermore, we demonstrate that IL‐15 overexpression can attenuate muscle fatigue associated with mammary tumour growth in a preclinical mouse model of breast cancer. Therefore, we propose that skeletal muscle fatigue is an inherent consequence of breast tumour growth, and this greater fatigue can be targeted therapeutically.
Aurora A kinase (AURKA) is overexpressed in 96% of human cancers and is considered an independent marker of poor prognosis. While the majority of tumors have elevated levels of AURKA protein, few ...have AURKA gene amplification, implying that posttranscriptional mechanisms regulating AURKA protein levels are significant. Here, we show that NEDD9, a known activator of AURKA, is directly involved in AURKA stability. Analysis of a comprehensive breast cancer tissue microarray revealed a tight correlation between the expression of both proteins, significantly corresponding with increased prognostic value. A decrease in AURKA, concomitant with increased ubiquitination and proteasome-dependent degradation, occurs due to depletion or knockout of NEDD9. Reexpression of wild-type NEDD9 was sufficient to rescue the observed phenomenon. Binding of NEDD9 to AURKA is critical for AURKA stabilization, as mutation of S296E was sufficient to disrupt binding and led to reduced AURKA protein levels. NEDD9 confers AURKA stability by limiting the binding of the cdh1-substrate recognition subunit of APC/C ubiquitin ligase to AURKA. Depletion of NEDD9 in tumor cells increases sensitivity to AURKA inhibitors. Combination therapy with NEDD9 short hairpin RNAs and AURKA inhibitors impairs tumor growth and distant metastasis in mice harboring xenografts of breast tumors. Collectively, our findings provide rationale for the use of AURKA inhibitors in treatment of metastatic tumors and predict the sensitivity of the patients to AURKA inhibitors based on NEDD9 expression.
Abstract
Background
Language deficits are cardinal manifestations of some frontotemporal dementia (FTD) phenotypes and also increasingly recognized in sporadic and familial amyotrophic lateral ...sclerosis (ALS). They have considerable social and quality‐of‐life implications, and adaptive strategies are challenging to implement. While the neuropsychological profiles of ALS–FTD phenotypes are well characterized, the neuronal underpinnings of language deficits are less well studied.
Methods
A multiparametric, quantitative neuroimaging study was conducted to characterize the involvement of language‐associated networks, tracts, and cortical regions with a panel of structural, diffusivity, and functional magnetic resonance imaging (MRI) metrics. Seven study groups were evaluated along the ALS–FTD spectrum: healthy controls (HC), individuals with ALS without cognitive impairment (ALSnci),
C9orf72
‐negative ALS–FTD,
C9orf72
‐positive ALS–FTD, behavioral‐variant FTD (bvFTD), nonfluent variant primary progressive aphasia (nfvPPA), and semantic variant PPA (svPPA). The integrity of the Broca's area, Wernicke's area, frontal aslant tract (FAT), arcuate fascicle (AF), inferior occipitofrontal fascicle (IFO), inferior longitudinal fascicle (ILF), superior longitudinal fascicle (SLF), and uncinate fascicle (UF) was quantitatively evaluated. The functional connectivity (FC) between Broca's and Wernicke’ areas and FC along the FAT was also specifically assessed.
Results
Patients with nfvPPA and svPPA exhibit distinctive patterns of gray and white matter degeneration in language‐associated brain regions. Individuals with bvFTD exhibit Broca's area, right FAT, right IFO, and UF degeneration. The ALSnci group exhibits Broca's area atrophy and decreased FC along the FAT. Both ALS–FTD cohorts, irrespective of
C9orf72
status, show bilateral FAT, AF, and IFO pathology. Interestingly, only
C9orf72
‐negative ALS–FTD patients exhibit bilateral uncinate and right ILF involvement, while
C9orf72
‐positive ALS–FTD patients do not.
Conclusions
Language‐associated tracts and networks are not only affected in language‐variant FTD phenotypes but also in ALS and bvFTD. Language domains should be routinely assessed in ALS irrespective of the genotype.
Digital polymerase chain reaction (dPCR) has the potential to enable accurate quantification of target DNA copy number provided that all target DNA molecules are successfully amplified. Following ...duplex dPCR analysis from a linear DNA target sequence that contains single copies of two independent template sequences, we have observed that amplification of both templates in a single partition does not always occur. To investigate this finding, we heated the target DNA solution to 95 °C for increasing time intervals and then immediately chilled on ice prior to preparing the dPCR mix. We observed an exponential decline in estimated copy number (R(2)≥ 0.98) of the two template sequences when amplified from either a linearized plasmid or a 388 base pair (bp) amplicon containing the same two template sequences. The distribution of amplifiable templates and the final concentration (copies per µL) were both affected by heat treatment of the samples at 95 °C from 0 s to 30 min. The proportion of target sequences from which only one of the two templates was amplified in a single partition (either 1507 or hmg only) increased over time, while the proportion of target sequences where both templates were amplified (1507 and hmg) in each individual partition declined rapidly from 94% to 52% (plasmid) and 88% to 31% (388 bp amplicon) suggesting an increase in number of targets from which both templates no longer amplify. A 10 min incubation at 95 °C reduced the initial amplifiable template concentration of the plasmid and the 388 bp amplicon by 59% and 91%, respectively. To determine if a similar decrease in amplifiable target occurs during the default pre-activation step of typical PCR amplification protocol, we used mastermixes with a 20 s or 10 min hot-start. The choice of mastermix and consequent pre-activation time did not affect the estimated plasmid concentration. Therefore, we conclude that prolonged exposure of this DNA template to elevated temperatures could lead to significant bias in dPCR measurements. However, care must be taken when designing PCR and non-PCR based experiments by reducing exposure of the DNA template to sustained elevated temperatures in order to improve accuracy in copy number estimation and concentration determination.
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