Högskolan i Skövde

Background: When designing, implementing, and evaluating a work site health promotion program, it is necessary to ensure that the program is evidence based. The present article aims to present in-depth information on the design of a longitudinal randomized controlled complementary intervention pilot study that follows the Consort recommendations to evaluate possible effects of a health promotive intervention in healthy workers.

Methods: Employees from four different workplaces were randomly assigned to one of the following groups: i) Massage and mental training (sitting in the armchair and receiving mechanical massage while listening to mental training programs, n=19), ii) Massage (sitting in the armchair and receiving mechanical massage only, n=19), iii) Mental training (sitting in the armchair and listening to mental training programs only, n=19), iv) Pause (sitting in the armchair but not receiving mechanical massage or listening to mental training programs, n=19), v) Control (not sitting in the armchair at all, n=17). The study lasted for eight weeks. Immediately before the randomization, after four weeks and after eight weeks the participants responded to statements from the Swedish Scale of Personality and had their heart rate, blood pressure and fingertip temperature measured.

Results: Receiving mechanical massage and listening to mental training programs, either separately or in combination, during working hours had some positive effects on the employees’ own evaluation of their health, as well as their heart rate, blood pressure and fingertip temperature. However, the intervention need to be evaluated further.

Conclusion: The approach described makes it possible to design, implement and evaluate a work site health promotion program, also on pilot-study level and these results should be seen as a first step towards larger randomized studies. This types of studies need to focus on healthy participants and special care should be taken to guarantee adequately powered study groups and their homogeneity.

The incidence of pancreatic ductal adenocarcinoma (PDAC) is increasing worldwide. The dismal prognosis and lack of effective treatments urges for increased research efforts in developing new treatment regimens. Since the development of new cancer treatments are expensive and time consuming, repurposing drugs is preferable when possible. This strategy will also be of great value for people in low income countries, to increase the availability of effective cancer treatments to affordable costs. Digitoxin, a cardiac glycoside, has been shown to have anti-cancer effects. It binds to the α subunit of the Na+/K+-ATPase, leading to increased concentrations of intracellular calcium and eventually cell death. There seems also to be other mechanisms elicited in cancer cells by digitoxin. The α subunit occurs in three isoforms, of which α3 has the highest affinity to digitoxin and frequently found over-expressed in tumor cells compared to normal cells.

PDAC cancer cells, both cell lines and tumors, differ in their genotype and in the metabolic subtype, proliferation rate and inflammatory status. To be able to individualize treatment regimens it is important to be aware of the specific vulnerabilities (genotypic or phenotypic characteristics increasing the sensitivity to digitoxin) of each PDAC tumor/cell line. This research aims to investigate the potential of using digitoxin as an anti-cancer treatment in PDAC, and analyze its effects on cell viability, metabolism and inflammatory status in PDAC cell lines in vitro with the goal to find biomarkers for digitoxin sensitivity.

The analyses of the effects of digitoxin was performed in five cell lines derived from PDAC tumors, either from primary tumors or metastases. Cell lines derived from PDAC are sensitive to digitoxin treatment to different degrees. High expression of α3 seems to be indicative for digitoxin sensitivity, as do a high proliferation rate seen in cell lines derived from primary tumors. Both subunit expression and proliferation rate should be further evaluated in PDAC tumors to confirm their potential to be used as biomarkers clinically.

In the hunt for the working mechanism behind the anti-cancer effects of digitoxin, the choline pathway, a pathway commonly affected in tumors was enlightened in the metabolomics study, affected in all five cell lines tested. Choline metabolites are important for maintaining the cell membrane and are involved in energy metabolism using lipids. Digitoxin induced an up-regulation of choline and glycerophosphocholine, which rendered us to propose a novel theory about possible interactions between two functional complexes in the cell membrane, the Na+/K+-ATPase/EGFR/c-Src and the EGFR/c-Src – Chkα. The hypothesis is that when digitoxin binds to the Na+/K+-ATPase it leads to inactivation of Chkα with a subsequent decrease in the synthesis of phosphocholine and phosphatidylcholine. Since cancer cells rely on abundance of choline metabolites, we believe an inhibition of this pathway to be deleterious for these cells.

Finally, we conclude that digitoxin has great potential as an anti-cancer treatment for some patients with pancreatic ductal adenocarcinoma. To optimize treatment results, a thorough investigation of the tumor genotype and phenotype must be done for each patient. To further increase treatment success, combination of digitoxin with other treatments for synergistic effects could be beneficial. 

Digitoxin has repeatedly shown to have negative effects on cancer cell viability; however, the actual mechanism is still unknown. In this study, we investigated the effects of digitoxin (1-100 nM) in four pancreatic cancer cell lines, BxPC-3, CFPAC-1, Panc-1, and AsPC-1. The cell lines differ in their KRAS/BRAF mutational status and primary tumor or metastasis origin. We could detect differences in the basal rates of cell proliferation, glycolysis, and ROS production, giving the cell lines different phenotypes. Digitoxin treatment induced apoptosis in all four cell lines, but to different degrees. Cells derived from primary tumors (Panc-1 and BxPC-3) were highly proliferating with a high proportion of cells in the S/G2 phase, and were more sensitive to digitoxin treatment than the cell lines derived from metastases (CFPAC-1 and AsPC-1), with a high proportion of cells in G0/G1. In addition, the effects of digitoxin on the rate of glycolysis, ROS production, and proliferation were dependent on the basal metabolism and origin of the cells. The KRAS downstream signaling pathways were not altered by digitoxin treatment, thus the effects exerted by digitoxin were probably disconnected from these signaling pathways. We conclude that digitoxin is a promising treatment in highly proliferating pancreatic tumors.

The alpha subunits (ATP1A1-3) of Na⁺/K⁺-ATPase binds digitoxin with varying affinity. The expression levels of these subunits dictate the anticancer effects of digitoxin. In the present study, three pancreatic cancer cell lines, AsPC-1, Panc-1 and CFPAC-1, were used to investigate the effects of digitoxin in relation to the expression of the subunits ATP1A1 and ATP1A3. Cell viability and intracellular calcium concentrations was measured in relation to the gene and protein expression of ATP1A1 and ATP1A3. Digitoxin was used to treat the cells at concentrations of 1-100 nM, and the intracellular calcium concentrations increased in a concentration-dependent manner in the Panc-1 and in the CFPAC-1 cells with treatment at 100 nM. In the AsPC-1 cells only the supraphysiological concentration of digitoxin (100 nM) resulted in a decrease in the number of viable cells (unviable cells increased to 22%), whereas it had no effect on intracellular calcium levels. The number of viable Panc-1 and CFPAC-1 cells decreased after digitoxin treatment at 25-100 nM (unviable Panc-1 cells increased to 33-59%; unviable CFPAC-1 cells increased to 22-56%). Digitoxin treatment also affected the transcriptional expression of the ATP1A1 and ATP1A3 subunits. In Panc-1 cells, ATP1A3 gene expression was negatively associated with the digitoxin concentration (25-100 nM). In the AsPC-1 and CFPAC-1 cells, the expression of the ATP1A1 gene increased in the cells treated with the 100 nM digitoxin concentration. The protein expression of ATP1A1 and ATP1A3 was not altered with digitoxin treatment. The basal protein expression of ATP1A1 was high in the AsPC-1 and CFPAC-1 cells, compared to the Panc-1 cells, in contrast to the basal expression of ATP1A3, which was higher in the Panc-1 cells, compared to the other pancreatic cancer cells used. On the whole, the present study demonstrates that the high expression of ATP1A3 renders pancreatic cancer cells more susceptible to digitoxin-induced cell death. The findings suggest that the expression of ATP1A3 may be used as a marker for tumor sensitivity to digitoxin treatment, where a high expression of ATP1A3 is favorable for the anticancer effects of digitoxin.

The prevalence of Chlamydia trachomatis in Sweden is well known, whereas the prevalence of Mycoplasma genitalium is less well documented. Youth clinics offer free contraception advice, sexually transmitted infection (STI) testing and/or contact tracing for the age group 15–25 years. The main objective of this study was to determine the prevalence of STIs, the presence of symptoms and the role of contact tracing. From July 2013 to March 2014, 1001 persons, 509 women and 492 men, were included in this study of six youth clinics in the Region of Västra Götaland. Symptoms were registered and whether the patient was tested because of contract tracing. Collection of urine samples, testing, treatment and disease registration were performed according to clinical routines. Urine samples were analysed for C. trachomatis/N. gonorrhoeae on the Cobas 4800 system (Roche). M. genitalium was analysed by lab-developed PCR. Genital infection was present in 16.8%. The prevalence of M. genitalium was higher than for C. trachomatis (9.6% and 7.1%). Men with symptoms have a significantly higher relative risk for infection with M. genitalium or C. trachomatis compared to asymptomatic men, while there is no increase for women. Contact tracing is important since positive outcome has a high relative risk for both infections. The prevalence of M. genitalium was higher than C. trachomatis in this study population. Initial testing for both C. trachomatis and M. genitalium should at least be considered for young men presenting with symptoms of genital infection. In finding positive cases, contact tracing is of great importance. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.

PPFIBP1 encodes for the liprin-β1 protein, which has been shown to play a role in neuronal outgrowth and synapse formation in Drosophila melanogaster. By exome and genome sequencing, we detected nine ultra-rare homozygous loss-of-function variants in 16 individuals from 12 unrelated families. The individuals presented with moderate to profound developmental delay, often refractory early-onset epilepsy, and progressive microcephaly. Further common clinical findings included muscular hyper- and hypotonia, spasticity, failure to thrive and short stature, feeding difficulties, impaired vision, and congenital heart defects. Neuroimaging revealed abnormalities of brain morphology with leukoencephalopathy, ventriculomegaly, cortical abnormalities, and intracranial periventricular calcifications as major features. In a fetus with intracranial calcifications, we identified a rare homozygous missense variant that by structural analysis was predicted to disturb the topology of the SAM domain region that is essential for protein-protein interaction. For further insight into the effects of PPFIBP1 loss of function, we performed automated behavioral phenotyping of a Caenorhabditis elegans PPFIBP1/hlb-1 knockout model, which revealed defects in spontaneous and light-induced behavior and confirmed resistance to the acetylcholinesterase inhibitor aldicarb, suggesting a defect in the neuronal presynaptic zone. In conclusion, we establish bi-allelic loss-of-function variants in PPFIBP1 as a cause of an autosomal recessive severe neurodevelopmental disorder with early-onset epilepsy, microcephaly, and periventricular calcifications. 

Objective We report 3 siblings with the characteristic features of ataxia-telangiectasia-like disorder associated with a homozygous MRE11 synonymous variant causing nonsense-mediated mRNA decay (NMD) and MRE11A deficiency. Methods Clinical assessments, next-generation sequencing, transcript and immunohistochemistry analyses were performed. Results The patients presented with poor balance, developmental delay during the first year of age, and suffered from intellectual disability from early childhood. They showed oculomotor apraxia, slurred and explosive speech, limb and gait ataxia, exaggerated deep tendon reflex, dystonic posture, and mirror movement in their hands. They developed mild cognitive abilities. Brain MRI in the index case revealed cerebellar atrophy. Next-generation sequencing revealed a homozygous synonymous variant in MRE11 (c.657C>T, p.Asn219=) that we show affects splicing. A complete absence of MRE11 transcripts in the index case suggested NMD and immunohistochemistry confirmed the absence of a stable protein. Conclusions Despite the critical role of MRE11A in double-strand break repair and its contribution to the Mre11/Rad50/Nbs1 complex, the absence of MRE11A is compatible with life. 

Neuroblastoma constitutes roughly 8% of all childhood cancers where 95% of all neuroblastoma cases occur before the age of 10. The survival rate of infants and young children is very poor, which alone contributes to research novel biomarkers for classification methods, improved diagnosis and better anti-tumor therapies. The aim of this meta-analysis was to identify dysregulated miRNAs in neuroblastoma that has the potential to be used as antioncogenic biomarkers for diagnostic interventions. Additionally, explore miRNA interconnectedness on a systemic level and conversely extend the support of using miRNAs as biomarkers. A comprehensive literature search was performed within NIH-PubMed, NCBI-PMC and in the reference list of already reviewed publications, which yielded 9 eligible publications. Quality of evidence was assessed according to the guidelines adapted from MIAME, MINSEQE and MIQE. miRNet 2.0 was used to find the most significantly enriched annotations linked to neuroblastoma. A total of 251 samples (Cancer: 141; Control: 110) was reported by the 9 studies. These involved 66 dysregulated miRNAs (Up-regulated: 43; Down-regulated: 23) which was used for enrichment analysis. Four miRNAs (miR-17-5p, -92a-3p -421, -125b) were significantly linked to neuroblastoma, and associated secondary diseases; medulloblastoma (-92a-3p, -125b), bladder cancer (-17-5p, -125b), acute myeloid leukemia (-92a-3p, -125b) and cardiac hypertrophy (- 125b). miR-125b showed exceptional interconnectivity with these diseases and a multidimensional potential in neural tumorigenesis. This study showed that dysregulation and biological processes of these miRNAs were concurrent with the original studies, endorsing that these miRNAs have potential as diagnostic indicators or classifiers of such diseases.