"Future patient II" telerehabilitation for patients with heart failure: Protocol for a randomized controlled trial.

Background: Heart failure is a global problem affecting millions of people worldwide. Current care of heart failure patients follows standard protocols and often overlooks the patients' specific needs, which leads to low compliance in the rehabilitation phase. Telerehabilitation, where the patients communicate with health care professionals about their rehabilitation program and monitor their vital signs, aims to increase the degree of compliance as well as enhancing their quality of life. Objective: The aim of this study is to investigate whether application of the Future Patient Telerehabilitation Program II can improve the health-related quality of life for patients with heart failure. Methods: A randomized controlled trial will be conducted. A total of 70 patients will be enrolled, 35 in the intervention group, 35 in the control group. The intervention group will follow an add-on to traditional care, while the control group will follow the conventional Danish cardiac rehabilitation program, which is based on periodic visits to the clinic. The patients will be followed for a period of six months. The intervention group will have access to an online HeartPortal and will use various home-based devices for self-monitoring. The primary outcome to be investigated is health-related quality of life as measured by the EuroQol-5 Dimension. Secondary outcomes are the number of visits to the outpatient clinic, number of readmissions and number of tele-communications contacts (phone and video) with health care professionals. The primary and secondary outcomes will be assessed using questionnaires and through the data generated by digital technologies for self-monitoring. Results: Enrolment began in August 2020. The results will be published in peer-reviewed journals. Results from the Future Patient II Telerehabilitation program are expected to be published in 2024. Discussion: This study is a further development of the Future Patient Telerehabilitation I study, and it is expected to explore the use of video consultation and a weight calculator in relation to telerehabilitation as well as the quality of life for heart failure patients. Conclusion: The expected outcomes are increased quality of life, increased number of phone- and video-consultations with health-care professionals, and the enhanced ability of patients to manage their own disease with the use of a calculator for weight.

NK/T-cell lymphoma.

In this case report, a 61-year-old male presented with odynophagia and ulceration in palatum durum after inhalating dust from machinery containing a weak acid. It was at first diagnosed as an acidic ulcer due to two biopsies verifying this. Because of progressing ulceration a third biopsy was taken - this time with the diagnosis extranodal NK/T-cell lymphoma, nasal type. This illustrates the diagnostic challenges of the illness, typically requiring multiple biopsies, and one should have this differential diagnosis in mind in case of progressing ulceration.

The role of aging and brain-derived neurotrophic factor signaling in expression of base excision repair genes in the human brain.

DNA damage is a central contributor to the aging process. In the brain, a major threat to the DNA is the considerable amount of reactive oxygen species produced, which can inflict oxidative DNA damage. This type of damage is removed by the base excision repair (BER) pathway, an essential DNA repair mechanism, which contributes to genome stability in the brain. Despite the crucial role of the BER pathway, insights into how this pathway is affected by aging in the human brain and the underlying regulatory mechanisms are very limited. By microarray analysis of four cortical brain regions from humans aged 20-99 years (n = 57), we show that the expression of core BER genes is largely downregulated during aging across brain regions. Moreover, we find that expression of many BER genes correlates positively with the expression of the neurotrophin brain-derived neurotrophic factor (BDNF) in the human brain. In line with this, we identify binding sites for the BDNF-activated transcription factor, cyclic-AMP response element-binding protein (CREB), in the promoter of most BER genes and confirm the ability of BDNF to regulate several BER genes by BDNF treatment of mouse primary hippocampal neurons. Together, these findings uncover the transcriptional landscape of BER genes during aging of the brain and suggest BDNF as an important regulator of BER in the human brain.

Developmental expression of CREB1 and NFATC2 in pig embryos.

BACKGROUND: The CREB1 gene encodes the cAMP response element binding protein 1 (CREB1), a leucine zipper transcription factor that regulates cellular gene expression in response to elevated levels of intracellular cAMP. When activated by phosphorylation, CREB1 binds to the cAMP response element (CRE) of the promoters of its target genes. CREB1 is an essential component in many physiological processes, and its function is correlated to neurodevelopment, plasticity and cell survival, and learning and memory. The NFATC2 gene codes for the nuclear factor of activated T-cells 2 protein. The NFATC2 protein is a DNA-binding protein that functions as an inducer of gene transcription during immune response. METHODS AND RESULTS: The aim of the present study was to examine the developmental expression of porcine CREB1 and NFACT2 transcripts. The expression of CREB1 and NFACT2 mRNA was examined by quantitative real-time RT-PCR. For the CREB1 transcript, we found significant reduction in transcript levels in the brain stem and basal ganglia during porcine embryo development, determined from day 60 to day 115 of gestation. In contrast, a significant increase in CREB1 mRNA was detected in the lungs during embryo development. No significant changes in the NFATC2 transcript were detected in porcine brain tissue during embryo development. CONCLUSIONS: Differential CREB1 mRNA expression was found in pig brain tissues during embryo development.

Bowhead NEIL1: molecular cloning, characterization, and enzymatic properties.

Nei Like DNA Glycosylase 1 (NEIL1) is a DNA glycosylase, which specifically processes oxidative DNA damage by initiating base excision repair. NEIL1 recognizes and removes bases, primarily oxidized pyrimidines, which have been damaged by endogenous oxidation or exogenous mutagenic agents. NEIL1 functions through a combined glycosylase/AP (apurinic/apyrimidinic)-lyase activity, whereby it cleaves the N-glycosylic bond between the DNA backbone and the damaged base via its glycosylase activity and hydrolysis of the DNA backbone through beta-delta elimination due to its AP-lyase activity. In our study we investigated our hypothesis proposing that the cancer resistance of the bowhead whale can be associated with a better DNA repair with NEIL1 being upregulated or more active. Here, we report the molecular cloning and characterization of three transcript variants of bowhead whale NEIL1 of which two were homologous to human transcripts. In addition, a novel NEIL1 transcript variant was found. A differential expression of NEIL mRNA was detected in bowhead eye, liver, kidney, and muscle. The A-to-I editing of NEIL1 mRNA was shown to be conserved in the bowhead and two adenosines in the 242Lys codon were subjected to editing. A mass spectroscopy analysis of liver and eye tissue failed to demonstrate the existence of a NEIL1 isoform originating from RNA editing. Recombinant bowhead and human NEIL1 were expressed in E. coli and assayed for enzymatic activity. Both bowhead and human recombinant NEIL1 catalyzed, with similar efficiency, the removal of a 5-hydroxyuracil lesion in a DNA bubble structure. Hence, these results do not support our hypothesis but do not refute the hypothesis either.

Exposure of pigs to glyphosate affects gene-specific DNA methylation and gene expression.

Glyphosate (N-(phosphonomethyl)glycine) is a broad-spectrum systemic herbicide and crop desiccant. Glyphosate has long been suspected of leading to the development of cancer and of compromising fertility. Herbicides have been increasingly recognized as epigenetic modifiers, and the impact of glyphosate on human and animal health might be mediated by epigenetic modifications. This article presents the results from an animal study where pigs were exposed to glyphosate while feeding. The experimental setup included a control group with no glyphosate added to the feed and two groups of pigs with 20 ppm and 200 ppm of glyphosate added to the feed, respectively. After exposure, the pigs were dissected, and tissues of the small intestine, liver, and kidney were used for DNA methylation and gene expression analyses. No significant change in global DNA methylation was found in the small intestine, kidney, or liver. Methylation status was determined for selected genes involved in various functions such as DNA repair and immune defense. In a CpG island of the promoter for IL18, we observed significantly reduced DNA methylation for certain individual CpG positions. However, this change in DNA methylation had no influence on IL18 mRNA expression. The expression of the DNA methylation enzymes DNMT1, DNMT3A, and DNMT3B was measured in the small intestine, kidney, and liver of pigs exposed to glyphosate. No significant changes in relative gene expression were found for these enzymes following dietary exposure to 20 and 200 ppm glyphosate. In contrast, a significant increase in expression of the enzyme TET3, responsible for demethylation, was observed in kidneys exposed to 200 ppm glyphosate.

[Assessment of nasal function by computational fluid dynamics].

This review summarises the knowledge of computational fluid dynamics (CFD) which combines fluid mechanics, mathematics and computer simulation to analyse airflow and air conditioning of the nasal airway. Traditional objective measures are hampered by low correlation to subjective outcome, whereas CFD variables such as heat flux and nasal middle airflow show good correlation. Studies also show great potential for virtual surgery, when the nasal procedure is simulated, and CFD analyse the impact on airflow and conditioning to optimize surgical planning. CFD could be of great value in rhinology and improve nasal surgery.

Health locus of control in a pandemic situation: cross-cultural differences between European and Asian respondents.

BACKGROUND: The present cross-cultural study examined the health locus of control construct during the COVID-19 pandemic. The scientific purpose of the study was to determine whether, during the pandemic situation, cultural and sex differences influence the health locus of control construct and change the internal health locus of control (IHLC), powerful others health locus of control (PHLC), and chance health locus of control (CHLC). PARTICIPANTS AND PROCEDURE: A total of 2617 recipients aged 18-70 years from Asia (China, India, and Indonesia), and Europe (Bulgaria, Germany, and Hungary) completed a questionnaire about their health. The participants completed an online version of the Multidimensional Health Locus of Control Scale - Form A. RESULTS: The survey shows that in a pandemic life-threatening situation, most individuals strive to rely on IHLC and/or PHLC, and fewer of them tend to rely on CHLC. However, there are differences (p < .001) between the two cultural samples: the representatives of Asian collectivistic culture are more dominated by PHLC, compared to the representatives of the European individualistic culture. When the comparison is between individuals from different cultures, sex differentiation affects the health locus of control, and as a result, significant differences in relation to IHLC, PHLC, and CHLC levels (p < .05) appear. CONCLUSIONS: In conclusion, the study indicates that cultural differences influence both the IHLC and PHLC levels, and that Asian participants are dominated by PHLC more than European respondents. Asian females are more likely to seek support from powerful others (PHLC) compared to European women, who perceive themselves as more independent. Asian male participants are prepared to rely on powerful others (doctors or medical institutions), while European male respondents are prone to rely on themselves mainly (IHLC). The results show that sex differences do not significantly affect the health locus of control within the same cultural group.

Workaholism across European and Asian cultures during the COVID-19 pandemic.

The current cross-cultural study examined the construct of workaholism across European and Asian cultures during the pandemic caused by coronavirus disease 2019 (COVID-19). A total of 2,617 recipients, aged 18-80 years from three Asian countries (China, India, and Indonesia) with higher levels of collectivistic values, and three European countries (Bulgaria, Germany, and Hungary) supposing to have higher individualistic values. The participants completed the online version of the two-dimensional measure, dubbed the Dutch Workaholism Scale (DUWAS). The goal of the study was to demonstrate that during the COVID-19 pandemic, it is the cultural context that mediates and influences the way of change in workaholics' attitudes. The results led to the conclusion that the way in which the COVID-19 crisis affects workaholism and workaholics' behavior depends on cultural and sex differences, and stages of the human life cycle. The data analysis revealed that cultural differences and sex affect the configuration of workaholism (excessive/compulsive): in the Asian sample, unlike the European, there was a significant increase in the level of workaholism compulsive; European female participants reported higher levels of workaholism compulsive and workaholism excessive, but the sex difference was not found in Asian sample. Along with cultural context, and sex differences, age also influences the configuration of workaholism. In this case, the separate stages of the human life cycle contribute in different ways to changes in levels of workaholism excessive and workaholism compulsive.

Conservation of A-to-I RNA editing in bowhead whale and pig.

RNA editing is a post-transcriptional process in which nucleotide changes are introduced into an RNA sequence, many of which can contribute to proteomic sequence variation. The most common type of RNA editing, contributing to nearly 99% of all editing events in RNA, is A-to-I (adenosine-to-inosine) editing mediated by double-stranded RNA-specific adenosine deaminase (ADAR) enzymes. A-to-I editing at 'recoding' sites results in non-synonymous substitutions in protein-coding sequences. Here, we present studies of the conservation of A-to-I editing in selected mRNAs between pigs, bowhead whales, humans and two shark species. All examined mRNAs-NEIL1, COG3, GRIA2, FLNA, FLNB, IGFBP7, AZIN1, BLCAP, GLI1, SON, HTR2C and ADAR2 -showed conservation of A-to-I editing of recoding sites. In addition, novel editing sites were identified in NEIL1 and GLI1 in bowhead whales. The A-to-I editing site of human NEIL1 in position 242 was conserved in the bowhead and porcine homologues. A novel editing site was discovered in Tyr244. Differential editing was detected at the two adenosines in the NEIL1 242 codon in both pig and bowhead NEIL1 mRNAs in various tissues and organs. No conservation of editing of KCNB1 and EEF1A mRNAs was seen in bowhead whales. In silico analyses revealed conservation of five adenosines in ADAR2, some of which are subject to A-to-I editing in bowheads and pigs, and conservation of a regulatory sequence in GRIA2 mRNA that is responsible for recognition of the ADAR editing enzyme.

The porcine cerebellin gene family.

Cerebellins (CBLN1-4), together with C1qTNF proteins, belong to the CBLN subfamily of C1q proteins. Cerebellin-1 (CBLN1) is active in synapse formation and functions at the parallel fiber-Purkinje cell synapses. Cerebellins form tripartite complexes with neurexins and the glutamate-receptor-related proteins GluD1 and GluD2, playing a role as trans-synaptic cell-adhesion molecules that critically contribute to both synapse formation and functioning and brain development. In this study, I present a molecular characterization of the four porcine CBLN genes. Experimental data and in silico analyses collectively describes the gene structure, chromosomal localization, and expression of CBLN1-4. Two cDNAs encoding the cerebellins CBLN1 and CBLN3 were RT-PCR cloned and sequenced. The nucleotide sequence of the CBLN1 clone contains an open reading frame of 582 nucleotides and encodes a protein of 193 amino acids. The deduced amino acid of the porcine CBLN1 protein was 99% identical to both mouse CBLN1 and to human CBLN1. The deduced CBLN1 protein contains a putative signal sequence of 21 residues, two conserved cysteine residues, and C1q domain. The nucleotide sequence of the CBLN3 cDNA clone comprises an open reading frame of 618 nucleotides and encodes a protein of 205 amino acids. The deduced amino acid sequence of the porcine CBLN3 protein was 88% identical to mouse CBLN3 and 94% identical to human CBLN3. The amino terminal ends of both the CBLN1 and CBLN3 proteins contain three possible N-linked glycosylation sites. The genomic organization of both porcine CBLN1 and CBLN3 is very similar to those of their human counterparts. The expression analyses demonstrated that CBLN1 and CBLN3 transcripts are predominantly expressed in the cerebellum. The sequences of the porcine precerebellin genes and cDNAs were submitted to DDBJ/EMBL/GenBank under the following accession numbers: CBLN1 gene (GenBank ID: FJ621565), CBLN1 cDNA (GenBank ID: EF577504), CBLN3 gene (GenBank ID: FJ621566), CBLN3 cDNA (GenBank ID: EF577505) and CBLN4 cDNA (GenBank ID: FJ196070).

Molecular characteristics of porcine alpha-synuclein splicing variants.

Alpha-synuclein (α-syn) is a 140 amino acid, intrinsically disordered protein with a potential role in neurotransmitter vesicle release. The protein is natively unfolded under physiological conditions, and is expressed predominantly in neural tissue. α-syn is associated with neuropathological conditions in Parkinson's disease, where the protein misfolds into oligomers and fibrils resulting in aggregates in Lewy bodies. Here we report the molecular cloning of SNCA cDNA encoding porcine α-syn and transcript variants hereof. Six transcripts coding for porcine α-syn are presented in the report, of which three result from exon skipping, generating in-frame splicing of coding exons 3 and 5. The splicing pattern of these alternative spliced variants is conserved between human and pig. All the observed in-frame deletions yield significantly shorter α-syn proteins compared with the 140 amino acid full-length protein. Expression analysis performed by real-time quantitative RT-PCR revealed a differential expression of the six transcript splicing variants in different pig organs and tissues. Common for all splicing variants, a very high transcript expression was detected in brain tissues and in spinal cord and very low or no expression outside the central nervous system. The porcine α-syn protein demonstrated markedly different biophysical characteristics compared with its human counterpart. No fibrillation of porcine α-syn was observed with the pig wild-type α-syn and A30P α-syn, and both variants show significantly reduced ability to bind to lipid vesicles. Overexpression of mutated porcine α-syn might recapitulate the human PD pathogenesis and lead to the identification of genetic modifiers of the disease.

Diagnostic accuracy of vascular ultrasound in patients with suspected giant cell arteritis (EUREKA): a prospective, multicentre, non-interventional, cohort study.

BACKGROUND: Temporal artery biopsy is considered the diagnostic gold standard for giant cell arteritis, despite approximately 39% of patients who are negative for the condition by biopsy subsequently being given a clinical diagnosis of giant cell arteritis. We aimed to assess the diagnostic accuracy of ultrasound examination in patients with suspected giant cell arteritis. METHODS: In this prospective, multicentre, non-interventional, cohort study (evaluation of ultrasound's role in patients suspected of having extracranial and cranial giant cell arteritis; EUREKA), we consecutively recruited patients aged 50 years or older, with clinically suspected giant cell arteritis from three Danish hospitals (South West Jutland Hospital in Esbjerg, Silkeborg Regional Hospital, and Rigshospitalet, Glostrup). Participants had a bilateral ultrasound of the temporal, facial, common carotid, and axillary arteries. Ultrasounds were done by ultrasonographers who were systematically trained in vascular ultrasound using appropriate equipment and settings. Participants then had a temporal artery biopsy within 7 days of initiation of corticosteroid treatment. A blinded ultrasound expert assessed all ultrasound images. Ultrasound vasculitis was defined in cranial arteries as a homogeneous, hypoechoic, intimamedia complex thickness and a positive compression sign and as a homogeneous intimamedia complex of 1 mm in thickness or wider in the axillary arteries and of 1·5 mm thickness or wider in the common carotid artery. Participants were followed up at 6 months. During this 6 month period, clinicians were able to collect data from all clinical examinations to enable a full clinical diagnosis at 6 months. Clinical diagnosis was based on the expert opinion of the treating rheumatologist. The diagnostic criterion standard was diagnosis confirmed after 6 months of follow-up. We used logistic regression analyses to calculate the odds ratio and 95% CI of ultrasound as a predictor for giant cell arteritis. FINDINGS: Between April 1, 2014, and July 31, 2017, 118 patients were screened for inclusion, of whom 106 had both ultrasound examinations and an eligible temporal artery biopsy and were included in the intention-to-diagnose population. The mean age was 72·7 years (SD 7·9), 63 (59%) participants were women, and 43 (41%) were men. Temporal artery biopsy was positive in 46 (43%) of 106 patients, and 62 (58%) of 106 patients had a clinically confirmed diagnosis of giant cell arteritis at 6 months (temporal artery biopsy sensitivity 74% [95% CI 62-84], specificity 100% [95% CI 92-100]). Cranial artery ultrasound was positive in all patients who had a positive temporal artery biopsy, and seven (58%) of 12 patients who were positive by ultrasound and negative by temporal artery biopsy were confirmed to have large-vessel giant cell arteritis via other imaging methods. The sensitivity of ultrasound diagnosis of giant cell arteritis was 94% (84-98) and specificity was 84% (70-93). Logistic regression analysis confirmed that ultrasound was the strongest baseline predictor for a clinically confirmed diagnosis of giant cell arteritis at 6 months (crude odds ratio 76·6 [95% CI 21·0-280·0]; adjusted for sex and age 141·0 [27·0-743·0]). INTERPRETATION: Vascular ultrasound might effectively replace temporal artery biopsy as a first-line diagnostic method in patients suspected of having giant cell arteritis when done by systematically trained ultrasonographers using appropriate equipment and settings. FUNDING: The Institute for Regional Research at Hospital of Southwest Jutland, Esbjerg, Denmark.

Non-Contact Respiratory Measurement Using a Depth Camera for Elderly People.

Measuring respiration at home for cardiac patients, a simple method that can detect the patient's natural respiration, is needed. The purpose of this study was to develop an algorithm for estimating the tidal volume (TV) and respiratory rate (RR) from the depth value of the chest and/or abdomen, which were captured using a depth camera. The data of two different breathing patterns (normal and deep) were acquired from both the depth camera and the spirometer. The experiment was performed under two different clothing conditions (undressed and wearing a T-shirt). Thirty-nine elderly volunteers (male = 14) were enrolled in the experiment. The TV estimation algorithm for each condition was determined by regression analysis using the volume data from the spirometer as the objective variable and the depth motion data from the depth camera as the explanatory variable. The RR estimation was calculated from the peak interval. The mean absolute relative errors of the estimated TV for males were 14.0% under undressed conditions and 10.7% under T-shirt-wearing conditions; meanwhile, the relative errors for females were 14.7% and 15.5%, respectively. The estimation error for the RR was zero out of a total of 206 breaths under undressed conditions and two out of a total of 218 breaths under T-shirt-wearing conditions for males. Concerning females, the error was three out of a total of 329 breaths under undressed conditions and five out of a total of 344 breaths under T-shirt-wearing conditions. The developed algorithm for RR estimation was accurate enough, but the estimated occasionally TV had large errors, especially in deep breathing. The cause of such errors in TV estimation is presumed to be a result of the whole-body motion and inadequate setting of the measurement area.

Vascular ultrasound for the diagnosis of giant cell arteritis: a reliability and agreement study based on a standardised training programme.

OBJECTIVE: To evaluate the impact of a standardised training programme including equipment adjustment for experienced musculoskeletal ultrasonographers without previous experience in vascular ultrasound (US) on the reliability of US in the diagnosis of giant cell arteritis (GCA). METHODS: In this prospective, non-interventional observational cohort study, patients suspected of GCA were evaluated by US by one of five rheumatologists with long-standing experience in musculoskeletal US (>8 years), trained using a standardised training programme including equipment adjustment. Images of cranial and large vessels were subsequently evaluated first by the performing ultrasonographer and thereafter by a blinded external expert (gold standard). RESULTS: In three Danish centres, 112 patients suspected of GCA were included. According to the external expert, vasculitis changes were seen in 66 patients, in 45 of them with only cranial involvement, in 14 with both cranial and large vessel involvement, while in seven patients isolated large vessel vasculitis was found. The reliability was excellent between the local ultrasonographer and the US expert for the overall GCA diagnosis regarding the diagnosis of cranial and for large vessel GCA, with an interobserver agreement of 95-96%, mean kappa values of 0.88-0.92 (95% CI 0.78 to 0.99). Excellent reliability (mean kappa 0.86-1.00) was also found for the US examination of the individual arteries (temporal, facial, common carotid and axillary). CONCLUSION: The US training programme resulted in excellent agreement between trainees and an expert in patients suspected of GCA and may thus be applicable for implementation of vascular US in clinical practice.

Combined in vitro fertilization and culture (IVF/IVC) in mouse for reprotoxicity assessment of xenobiotic exposure.

Litter size and other conventional measures in rodents are common end-points in the assessment of xenobiotics for reprotoxic effects. However, since litter size may be normal despite reduced semen quality, we established and tested a mouse in vitro fertilization/in vitro culture (IVF/IVC) system to assess other aspects of reprotoxicity of xenobiotic exposure. Two pesticides, vinclozolin (V) and chlormequat (C), were added to feed in low (40 and 900 ppm, respectively) and high (300 and 2700 ppm, respectively) doses and compared to control (nil pesticide). Exposed males were used for natural mating to evaluate litter size and then used for IVF/IVC and sperm evaluation. The IVF/IVC system detected significant adverse effect of high dose of vinclozolin on blastocyst formation, which was not detected by conventional measures such as litter size or sperm motility and viability. We conclude that assessment based on IVF/IVC measures may complement litter size and other conventional end-points.

Molecular cloning and characterization of porcine Na⁺/K⁺-ATPase isoform α4.

Na+/K+-ATPase is responsible for maintaining electrochemical gradients of Na+ and K+, which is essential for a variety of cellular functions including neuronal activity. The α-subunit of the Na+/K+-ATPase is composed of four different polypeptides (α1-α4) encoded by different genes. Na,K-ATPase α4, encoded by the ATP1A4 gene, is expressed in testis and in male germ cells of humans, rats and mice. The α4 polypeptide has an important role in sperm motility, and is essential for male fertility. Here we present the RT-PCR cloning and characterization of the porcine ATP1A4 cDNA coding for Na⁺/K⁺-ATPase polypeptide α4. The Na⁺/K⁺-ATPase polypeptide α4, consisting of 1030 amino acids, displays a high homology with its human counterpart (86%). Phylogenetic analysis demonstrated that porcine Na⁺/K⁺-ATPase polypeptide α4 is closely related to other mammalian counterparts. In addition, the genomic structure of the porcine ATP1A4 gene was determined, and the intron-exon organization was found to be similar to that of the human ATP1A4 gene. The promoter sequence for the porcine ATP1A4 gene was also identified. Investigation of the genetic variation in the porcine ATP1A4 gene revealed a missense A/G SNP in exon 18. This A/G polymorphism results in a substitution of a methionine to a glycine residue (M888G). A very high overall DNA methylation rate of the ATP1A4 gene, 70-80%, was observed in both brain and liver. Expression analysis demonstrated that the porcine ATP1A4 gene is predominantly expressed in testis. The sequence of the porcine ATP1A4 cDNA encoding the Na⁺/K⁺-ATPase α4 protein has been submitted to GenBank under the accession number GenBank Accession No. MG587082.

Calibration of sperm concentration for in vitro fertilization in a mouse reprotoxicity model.

Xenobiotics, such as chemicals and pesticides, may result in adverse effects on reproduction in human and animals. Using in-vitro embryo production as a testing system reveals details of fertilization (IVF) and early embryonic development (IVC). The aim of our study was to perform a systematical calibration of sperm concentration in an IVF/IVC system, using an outbred mouse strain, and further determine the sperm concentration that furnishes a sensitive assessment of sperm fertilizing capacity in relation to reprotoxic evaluations. By performing breakpoint analysis, the results revealed a maximum two-cell percentage (51%, 95% CI: 38 to 69%) at 3.6 × 104 sperm/ml (95% CI: 2.1 × 104 to 6.1 × 104). For future application of the IVF/IVC system, a sperm concentration lower than this breakpoint concentration is required to be within the responsive range for determining sperm fertilizing capacity. We conclude that a relatively low sperm concentration (2.5 × 104 sperm/ml) is a precondition in a mouse IVF/IVC system in order to detect potential reprotoxic effects on sperm fertilizing capacity. Our study illustrates that a systematic approach is necessary for validation and appropriate use of such in-vitro system used for reproductive toxicity testing.

α-Synucleins from Animal Species Show Low Fibrillation Propensities and Weak Oligomer Membrane Disruption.

The intrinsically disordered protein α-synuclein (aSN) forms insoluble aggregates in the brains of Parkinson's disease (PD) patients. Cytotoxicity is attributed to a soluble aSN oligomeric species that permeabilizes membranes significantly more than monomers and fibrils. In humans, the A53T mutation induces early onset PD and increases the level of aSN oligomerization and fibrillation propensity, but Thr53 occurs naturally in aSNs of most animals. We compared aSNs from elephant, bowhead whale, and pig with human aSN. While all three animal aSNs showed significantly weakened fibrillation, elephant aSN formed much more oligomer, and pig aSN much less, than human aSN did. However, all animal aSN oligomers showed weakened permeabilization toward anionic lipid vesicles, indicative of decreased cytotoxicity. These animal aSNs share three substitutions compared to human aSN: A53T, G68E, and V95G. We analyzed aggregation and membrane binding of all eight mutants combining these three mutations. While the G68E mutation is particularly important in weakening fibrillation and possible toxicity, the strongest effect is seen when all three mutations are present. Thus, a small number of mutations can significantly decrease aSN toxicity.

Sleep apnoea in patients undergoing bariatric surgery
.

INTRODUCTION: The objective of this study was to investigate the presence of obstructive sleep apnoea (OSA) among patients awaiting bariatric surgery and to evaluate if a change in symptoms and clinical measurements of OSA was seen one year after bariatric surgery.
 Methods: Patients awaiting bariatric surgery in the Region of Southern Denmark were invited to participate in an OSA examination during a 15-month period (2012-2013) using the Embletta device for cardiorespiratory monitoring before and one year after bariatric surgery. The Apnoea-Hypopnoea Index (AHI), weight, BMI and the Epworth Sleepiness Score (ESS) were measured prior to and one year after surgery. 
Results: A total of 56 patients were enrolled in the study, and 59% were found with OSA (AHI ≥ 5). Thirty-six patients were eligible for examination one year post-operatively. Twelve of these patients did not have OSA (AHI < 5) either at inclusion or at re-examination. In the remaining 24 patients with OSA, the BMI dropped from 44.4 prior to surgery to 30.8 kg/m2 one year after surgery (p < 0.01). Mean AHI decreased from 12.8 prior to surgery to 3.7 one year after surgery (p < 0.01). There was no effect of weight reduction on the ESS.
 Conclusions: A statistically significant reduction in AHI was seen in patients with OSA one year after surgery. No statistical differences were observed for ESS. 
Funding: This study was funded by "Fonden for Læge-videnskabelig Forskning m.v. ved sygehusene i Region Syd" and by "Edith og Vagn Hedegaard Jensen fond".
Trial registration: ClinGov (ID: S-20120004jln).