Effects of journey duration and temperature during pre-slaughter transport on behaviour of cull sows in lairage.

This observational field study investigated the effects of journey duration, temperature, and waiting duration before unloading on the behaviour of 562 cull sows during lairage from 23 commercial loads. Each load consisted of sows originating from more than one herd, thus experiencing variable pre-slaughter transport and management. In lairage, sows were mixed in groups of 25, involving animals from all journey durations (min-max: 0.8-8.4 h) and video monitored for 60 min. At first most sows were in upright position (approximately 80-90%), decreasing to 30-40% after 30 min. After 60 min, 42% of the sows had initiated aggression (min-max: 0-43 events/sow), 28% had been subjected to aggressive behaviour (min-max: 0-14 events/sow), and 36% s were observed drinking (min-max: 0-16 events/sow). Several significant interactions were found between journey duration, the average temperature in the vehicle and lairage pen (averages: 4.3-26.2 °C) and waiting duration before unloading (min-max: 3-25 min). For example, after short journeys, sows exposed to higher temperature carried out more aggressive behaviour, while a higher temperature after long journeys was associated with more lying and less drinking. This suggests that the sows prioritised lying behaviour over drinking and establishing a dominance hierarchy. We discuss how the results may be interpreted as behavioural signs of fatigue, but further studies, for example involving quantification of physiological and motivational indicators, are needed to clarify this. Irrespectively, the present findings suggest that a stay in a lairage pen, as part of the pre-slaughter logistic chain, involves challenges for the welfare of the cull sows.

Coping strategies and considerations regarding low anterior resection syndrome and quality of life among patients with rectal cancer; a qualitative interview study.

Introduction: Low anterior resection syndrome (LARS) is defined as disordered bowel function following rectal resection, which is detrimental to quality of life (QoL). A recent international consensus definition of LARS stresses the importance of focusing on both the symptoms and the consequences that the symptoms have for the individual patient as studies indicate that LARS has a negative impact on patients' QoL. However, an ongoing PROM study investigating late sequelae after rectal cancer finds that a minor proportion of patients scoring major LARS experience none or only little impact on quality of life. Aim: The aim of this study was to identify patients' considerations and coping strategies to establish why the burden caused by major LARS had little or no influence on their QoL. Materials and methods: This was a qualitative interview study based on 21 semi-structured individual telephone interviews with patients treated for rectal cancer. Data were analysed using a hermeneutic inspired thematic analysis. Results and conclusion: Three themes emerged from the analysis; Adapting new life situation, Altering life perception and the Importance of relationships. Major LARS and its consequences following rectal cancer may be managed or altered by adopting problem-focused and emotion-focused coping strategies. Maintaining a positive attitude and having a good network of family and friends constitute a surplus, allowing patients to cope with the need for changed behaviour and appreciate the life that they have been given. Accepting that major LARS and its consequences cause limitations in life allowed patients to change their normality threshold over time.

The gut hormone Allatostatin C/Somatostatin regulates food intake and metabolic homeostasis under nutrient stress.

The intestine is a central regulator of metabolic homeostasis. Dietary inputs are absorbed through the gut, which senses their nutritional value and relays hormonal information to other organs to coordinate systemic energy balance. However, the gut-derived hormones affecting metabolic and behavioral responses are poorly defined. Here we show that the endocrine cells of the Drosophila gut sense nutrient stress through a mechanism that involves the TOR pathway and in response secrete the peptide hormone allatostatin C, a Drosophila somatostatin homolog. Gut-derived allatostatin C induces secretion of glucagon-like adipokinetic hormone to coordinate food intake and energy mobilization. Loss of gut Allatostatin C or its receptor in the adipokinetic-hormone-producing cells impairs lipid and sugar mobilization during fasting, leading to hypoglycemia. Our findings illustrate a nutrient-responsive endocrine mechanism that maintains energy homeostasis under nutrient-stress conditions, a function that is essential to health and whose failure can lead to metabolic disorders.

Risk of deep vein thrombosis and pulmonary embolism after gynecological day surgery.

OBJECTIVE: To investigate the risk of venous thromboembolism (VTE) in Danish women operated within a day surgery setting and to evaluate whether the current use of thromboprophylaxis without using graduated elastic compression stockings (GCS) is an appropriate treatment to prevent VTE. STUDY DESIGN: A retrospective cohort study including women who underwent laparoscopic hysterectomy or vaginal prolapse operation for benign disease from January 2014 to December 2017 at the Gynecology Day Surgery Unit, Regional Hospital of Randers, Denmark. The primary outcome was VTE diagnosed within three months postoperatively. Only one dose of pharmacological thromboprophylaxis (PTP) was given to women stratified at high risk of VTE. None of the women used GCS. RESULTS: A total of 671 women were included. Vaginal prolapse operations were performed on 626 women, and laparoscopic hysterectomy on 45 women. PTP was used for only 220 (32.8%) of these women. A total of 346 (51.5%) women were stratified as at high risk of VTE according to the national recommendations. Only 218 (63%) of these women received PTP, while 128 women (37%) did not receive PTP. The incidence of VTE within three months postoperatively was 0%. Only 13 (1.9%) of the women were readmitted within 14 days postoperatively due to hemorrhaging or hematoma; six out of these 13 women (46%) received PTP postoperatively. Re-operation was performed in seven (1%) women due to hemorrhaging, and three out of the seven (42.9%) had PTP postoperatively. CONCLUSION: The risk of VTE in Danish women operated within a day surgery setting is probably very low since we found no cases of VTE in our setup. The beneficial effect of routine use of GCS and one dose of PTP postoperatively given to all women who had undergone MIS in a day surgery setting are questioned. One dose of PTP postoperatively without GCS can be considered to only women stratified as high-risk of VTE until there is more evidence whether these women actually need thromboprophylaxis postoperatively at all. PRECIS: The incidence of VTE in women undergoing laparoscopic hysterectomy or vaginal prolapse operation in a day surgery setting without using graduated elastic compression stockings is very low.

A cross-sectional analysis of persistent low back pain, using correlations between lumbar stiffness, pressure pain threshold, and heat pain threshold.

INTRODUCTION: Little is known about the underlying biomechanical cause of low back pain (LBP). Recently, technological advances have made it possible to quantify biomechanical and neurophysiological measurements, potentially relevant factors in understanding LBP etiology. However, few studies have explored the relation between these factors. This study aims to quantify the correlation between biomechanical and neurophysiological outcomes in non-specific LBP and examine whether these correlations differ when considered regionally vs. segmentally. METHODS: This is a secondary cross-sectional analysis of 132 participants with persistent non-specific LBP. Biomechanical data included spinal stiffness (global stiffness) measured by a rolling indenter. Neurophysiological data included pain sensitivity (pressure pain threshold and heat pain threshold) measured by a pressure algometer and a thermode. Correlations were tested using Pearson's product-moment correlation or Spearman's rank correlation as appropriate. The association between these outcomes and the segmental level was tested using ANOVA with post-hoc Tukey corrected comparisons. RESULTS: A moderate positive correlation was found between spinal stiffness and pressure pain threshold, i.e., high degrees of stiffness were associated with high pressure pain thresholds. The correlation between spinal stiffness and heat pain threshold was poor and not statistically significant. Aside from a statistically significant minor association between the lower and the upper lumbar segments and stiffness, no other segmental relation was shown. CONCLUSIONS: The moderate correlation between spinal stiffness and mechanical pain sensitivity was the opposite of expected, meaning higher degrees of stiffness was associated with higher pressure pain thresholds. No clinically relevant segmental association existed.

Evaluation of the in vitro effects of local anesthetics on equine chondrocytes and fibroblast-like synoviocytes.

OBJECTIVE: To investigate the in vitro effects of clinically relevant concentrations of the local anesthetics (LAs) bupivacaine, lidocaine, lidocaine with preservative (LP), mepivacaine, and ropivacaine on equine chondrocyte and fibroblast-like synoviocyte (FLS) viability. SAMPLES: Chondrocytes and FLSs of the metacarpophalangeal joints of 4 healthy adult horses. PROCEDURES: Viability of chondrocytes and FLSs was determined with 3 assays: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH), and trypan blue (TB) exclusion (only FLS). Viability was assessed after 30- and 60-minute exposures to 0.0625%, 0.125%, and 0.25% bupivacaine; 0.25%, 0.5%, and 1% lidocaine; 0.25%, 0.5%, and 1% LP; 0.25%, 0.5%, and 1% mepivacaine; and 0.125%, 0.25%, and 0.5% ropivacaine. RESULTS: Viability of chondrocytes was significantly decreased with exposure to 0.25% bupivacaine, 1% lidocaine, 1% LP, 1% mepivacaine, and 0.25% ropivacaine. Viability of FLSs was significantly decreased with exposure to 0.25% bupivacaine, 1% mepivacaine, 1% LP, and 0.5% ropivacaine. CONCLUSIONS AND CLINICAL RELEVANCE: Clinically relevant concentrations of LAs had in vitro time- and concentration-dependent cytotoxicity for chondrocytes and FLSs isolated from the metacarpophalangeal joints of healthy horses. Bupivacaine was more toxic to chondrocytes than lidocaine, mepivacaine, and ropivacaine, whereas bupivacaine, LP, mepivacaine, and ropivacaine were more toxic to FLSs than preservative-free lidocaine. Several LAs may negatively affect chondrocyte and FLS viability.

Vocational Rehabilitation in Mild Traumatic Brain Injury: Supporting Return to Work and Daily Life Functioning.

Persisting post-concussive symptoms are challenging to treat and may delay return-to-work (RTW). The aims of this study were to describe a multidisciplinary and holistic vocational rehabilitation (VR) program for individuals with mild traumatic brain injury (mTBI) and to explore course and predictors of employment outcome during VR. The VR program was described using the Standard Operating Procedures (SOPs) framework. Further, a retrospective, cohort study on individuals with mTBI receiving VR was conducted based on clinical records (n = 32; 22% males; mean age 43.2 years; 1.2 years since injury on average). The primary outcome was difference in hours at work per week from pre- to post-VR, and the secondary outcome was change in a three-level RTW-status. Time since injury, age, sex, and loss of consciousness were investigated as predictors of the outcomes. The VR intervention is individually tailored and targets patients' individual needs. Thus, it may combine a variety of methods based on a biopsychosocial theoretical model. During VR, hours at work, 17.0 ± 2.2, p < 0.001, and RTW-status, OR = 14.0, p < 0.001, improved significantly with 97% having returned to work after VR. Shorter length of time since injury and male sex were identified as predictors of a greater gain of working hours. Time since injury was the strongest predictor; double the time was associated with a reduction in effect by 4.2 ± 1.4 h after adjusting for working hours at start of VR. In sum, these results suggest that individuals facing persistent problems following mTBI may still improve employment outcomes and RTW after receiving this multidisciplinary and holistic VR intervention, even years after injury. While results are preliminary and subject to bias due to the lack of a control group, this study warrants further research into employment outcomes and VR following mTBI, including who may benefit the most from treatment.

[Drawing improves understanding of anatomy, operation planning and communication].

This review summarises the knowledge of drawing as a learning tool. Drawing is a skill, which can be used by doctors in communicating with their patients. It also seems, that drawing supplements traditional learning methods and improves learning and retention of knowledge. Drawing is used by some doctors for planning of operations and as a visual tool to explain procedures and improve communication with patients. New visual technologies like augmented reality, virtual reality and 3D printing seem promising when used for training surgical skills and anatomy or planning operations.

Cell-based therapy for the treatment of female stress urinary incontinence: an early cost-effectiveness analysis.

AIM: To perform an early cost-effectiveness analysis of in vitro expanded myoblasts (IVM) and minced myofibers versus midurethral slings (MUS) for surgical treatment of female stress urinary incontinence. METHODS: Cost-effectiveness and sensitivity analyses were performed using a decision tree comprising previously published data and expert opinions. RESULTS & CONCLUSION: In the base case scenario, MUS was the cost-effective strategy with a negative incremental cost-effectiveness ratio compared with IVM and a positive incremental cost-effectiveness ratio compared with minced myofibers. However, the sensitivity analysis indicates that IVM may become an alternative providing greater effect at a higher cost. With the possibility of becoming more effective, IVM treatment would be advantageous over MUS given its reduced invasiveness and lower risks of complications.

Capsaicin-sensitive cutaneous primary afferents convey electrically induced itch in humans.

Specially designed transcutaneous electrical stimulation paradigms can be used to provoke experimental itch. However, it is unclear which primary afferent fibers are activated and whether they represent pathophysiologically relevant, C-fiber mediated itch. Since low-threshold mechano-receptors have recently been implicated in pruriception we aimed to characterize the peripheral primary afferent subpopulation conveying electrically evoked itch in humans (50Hz stimulation, 100µs square pulses, stimulus-response function to graded stimulus intensity). In 10 healthy male volunteers a placebo-controlled, 24-h 8% topical capsaicin-induced defunctionalization of capsaicin-sensitive (transient receptor potential V1-positive, 'TRPV1'+) cutaneous fibers was performed. Histaminergic itch (1% solution introduced by a prick test lancet) was provoked as a positive control condition. Capsaicin pretreatment induced profound loss of warmth and heat pain sensitivity (pain threshold and supra-threshold ratings) as assessed by quantitative sensory testing, indicative of efficient TRPV1-fiber defunctionalization (all outcomes: P<0.0001). The topical capsaicin robustly, and with similar efficaciousness, inhibited itch intensity evoked by electrical stimulation and histamine (-89±4.1% and -78±4.9%, respectively, both: P<0.0001 compared to the placebo patch area). The predominant primary afferent substrate for electrically evoked itch in humans, using the presently applied stimulation paradigm, is concluded to be capsaicin-sensitive polymodal C-fibers.

MicroRNA Profiling in the Medial and Lateral Habenula of Rats Exposed to the Learned Helplessness Paradigm: Candidate Biomarkers for Susceptibility and Resilience to Inescapable Shock.

Depression is a highly heterogeneous disorder presumably caused by a combination of several factors ultimately causing the pathological condition. The genetic liability model of depression is likely to be of polygenic heterogeneity. miRNAs can regulate multiple genes simultaneously and therefore are candidates that align with this model. The habenula has been linked to depression in both clinical and animal studies, shifting interest towards this region as a neural substrate in depression. The goal of the present study was to search for alterations in miRNA expression levels in the medial and lateral habenula of rats exposed to the learned helplessness (LH) rat model of depression. Ten miRNAs showed significant alterations associating with their response to the LH paradigm. Of these, six and four miRNAs were significantly regulated in the MHb and LHb, respectively. In the MHb we identified miR-490, miR-291a-3p, MiR-467a, miR-216a, miR-18b, and miR-302a. In the LHb miR-543, miR-367, miR-467c, and miR-760-5p were significantly regulated. A target gene analysis showed that several of the target genes are involved in MAPK signaling, neutrophin signaling, and ErbB signaling, indicating that neurotransmission is affected in the habenula as a consequence of exposure to the LH paradigm.

Time from first symptom experience to help seeking for colorectal cancer patients: Associations with cognitive and emotional symptom representations.

OBJECTIVES: The aim was to assess the association between cognitive and emotional symptom representations prior to diagnosis and the length of the patient interval (i.e. the time from the first symptom is experienced until healthcare is sought) for colorectal cancer patients. METHOD: The study population included 436 newly diagnosed colorectal cancer patients. Questionnaire data were collected using the Danish Revised Illness Perception Questionnaire (IPQ-R), including cognitive and emotional symptom representations and information on the patient interval. RESULTS: High score in treatment control was associated with short patient interval (PR = 0.52, 95% CI: 0.31-0.89) and high score on the timeline cyclical dimension was associated with long patient interval (PR = 2.14, 95% CI: 1.29-3.57). Hence, patients with negative beliefs about the treatability of their symptoms and patients with strong beliefs about the cyclical nature of their symptoms were more likely to have a long patient interval. Assigning blood in stool as the most important symptom significantly interacted in the association between the patient interval and the two cognitive symptom representations consequence and personal control. CONCLUSION: The results indicate that aspects of symptom representations were associated with the patient's help-seeking. PRACTICAL IMPLICATIONS: These findings may help clinicians and public health planners shorten patient intervals.

Evaluation of the iPLEX® ADME PGx Pro Panel and allele frequencies of pharmacogenetic markers in Danes.

OBJECTIVES: The iPlex® ADME PGx Pro Panel was developed to investigate 191 polymorphisms including single nucleotide polymorphisms (SNPs), insertion-deletions (INDELS), and copy number variations (CNV) relevant for absorption, distribution, metabolism, and excretion (ADME) of drugs. The purpose of this study was to perform a technical evaluation of the iPlex® ADME PGx Pro Panel by genotyping 50 unrelated Danes and estimate preliminary genotype frequencies among Danes. DESIGN AND METHODS: The investigations were performed by the use of PCR, single base extension (SBE) and Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS). RESULTS: The typing quality of 161 SNP assays was categorized as well performing or acceptable, whereas 22 SNP assays were categorized as either questionable or unacceptable. The frequencies of the genotypes observed in the Danish population were compared to those of the European reference population from the 1000 Genome Project. Three SNPs (rs737865, rs35167514, and rs34305973) showed statistically significantly differences between the frequencies of the 1000 Genomes Europeans and the Danes. The CNV assays could only be used as a guideline. CONCLUSION: In conclusion, the iPlex® ADME PGx Pro Panel is a cost-effective way of genotyping genes relevant for ADME.

A natural frameshift mutation in Campanula EIL2 correlates with ethylene insensitivity in flowers.

BACKGROUND: The phytohormone ethylene plays a central role in development and senescence of climacteric flowers. In ornamental plant production, ethylene sensitive plants are usually protected against negative effects of ethylene by application of chemical inhibitors. In Campanula, flowers are sensitive to even minute concentrations of ethylene. RESULTS: Monitoring flower longevity in three Campanula species revealed C. portenschlagiana (Cp) as ethylene sensitive, C. formanekiana (Cf) with intermediate sensitivity and C. medium (Cm) as ethylene insensitive. We identified key elements in ethylene signal transduction, specifically in Ethylene Response Sensor 2 (ERS2), Constitutive Triple Response 1 (CTR1) and Ethylene Insensitive 3- Like 1 and 2 (EIL1 and EIL2) homologous. Transcripts of ERS2, CTR1 and EIL1 were constitutively expressed in all species both throughout flower development and in response to ethylene. In contrast, EIL2 was found only in Cf and Cm. We identified a natural mutation in Cmeil2 causing a frameshift which resulted in difference in expression levels of EIL2, with more than 100-fold change between Cf and Cm in young flowers. CONCLUSIONS: This study shows that the naturally occurring 7 bp frameshift discovered in Cmeil2, a key gene in the ethylene signaling pathway, correlates with ethylene insensitivity in flowers. We suggest that transfer of the eil2 mutation to other plant species will provide a novel tool to engineer ethylene insensitive flowers.

Psychiatric morbidity and non-participation in breast cancer screening.

BACKGROUND: Organised breast cancer screening is currently one of the best strategies for early-stage breast cancer detection. However, early detection has proven challenging for women with psychiatric disease. This study aims to investigate psychiatric morbidity and non-participation in breast cancer screening. METHODS: We conducted an observational cohort study including women invited to the first organised screening round in the Central Denmark Region. Data on psychiatric diagnosis, psychoactive prescription medicine and consultation with private psychiatrists were obtained from Danish registries and assessed for a period of up to 10 years before the screening date. RESULTS: The cohort comprised 144,264 women whereof 33.0% were registered with an indication of psychiatric morbidity. We found elevated non-participation propensity among women with a psychiatric diagnosis especially for women with schizophrenia and substance abuse. Also milder psychiatric morbidity was associated with higher non-participation likelihood as women who had redeemed psychoactive prescription medicine or have had minimum one consultation with a private psychiatrist were more likely not to participate. Finally, we found that the chronicity of psychiatric morbidity was associated with non-participation and that woman who had a psychiatric morbidity defined as 'persistent' had higher likelihood of non-participation than women with recently active morbidity or inactive psychiatric morbidity. CONCLUSION: This study showed a strong association between psychiatric morbidity and an increased likelihood of non-participation in breast cancer screening in a health care system with universal and tax-funded health services. This knowledge may inform interventions targeting women with psychiatric morbidity as they have poorer breast cancer prognosis.

Social support and non-participation in breast cancer screening: a Danish cohort study.

BACKGROUND: Social support may have an impact on screening participation. We studied the association between social support in 2006, defined as frequencies of contacts, instrumental support and emotional support and participation in breast cancer screening in 2008-09. METHODS: This population-based cohort study included 4512 women who had participated in a Health Survey in 2006 and who also were in the target group for the first round of organized breast cancer screening in the Central Denmark region in 2008-09. RESULTS: Women with infrequent contacts with friends and family in 2006 were more likely not to participate in screening in 2008-09 [prevalence ratio (PR) 1.69, 95% confidence interval (CI) 1.26-2.26, P-value < 0.001 and PR 1.56, 95% CI 1.21-2.20, P-value < 0.001, respectively] as were women who reported not to have someone to look after her home if she was away for some time and women who reported usually not or never having someone to turn to with personal concerns (PR 1.97, 95% CI 1.53-2.54, P-value < 0.001 and PR 1.42, 95% CI 1.14-1.77, P-value = 0.002, respectively). CONCLUSIONS: Low social support, indicated by items in each social support attribute, was associated with non-participation in breast cancer screening in 2008-09. Targeted social interventions may, therefore, have an impact on future screening behaviour, which calls for further research.

Self-assessed health, perceived stress and non-participation in breast cancer screening: A Danish cohort study.

OBJECTIVE: Population-based cancer screening is offered in many countries to detect early stages of cancer and reduce mortality. Screening efficiency and equality is susceptible due to a group of non-participants. We investigated associations between self-assessed health, perceived stress and subsequent non-participation in breast cancer screening. METHODS: This population-based cohort study included 4512 women who had participated in a Health Survey in 2006 and who were also the target group (aged 50-69 years) for the first organised breast cancer screening programme -3 years later in the Central Denmark Region in 2008-2009. RESULTS: A U-shaped association was observed for physical health assessment as women with the highest (PR=1.28, 95% CI: 1.06-1.55), and the lowest (PR=1.41, 95% CI: 1.18-1.68) physical health scores were less likely to participate in the programme than women with physical health scores in the middle range. Women with low mental health assessment were more likely not to participate than women with mental health scores in the middle range (PR=1.44, 95% CI: 1.22-1.69). Higher non-participation propensity was also observed for women with the highest perceived stress scores (PR=1.27, 95% CI: 1.07-1.51) compared with women scoring in the middle range. CONCLUSIONS: Women with highest and lowest self-assessed physical health, with lowest mental health or highest perceived stress were significantly more likely not to participate in breast cancer screening 2-3 years later than women who reported average health. Interventions targeting these groups may promote equal participation in future breast cancer screening programmes.

Symptom attributions in patients with colorectal cancer.

BACKGROUND: Symptoms of cancer may be interpreted differently by different patients before the diagnosis. This study investigated symptom attributions in Danish patients with colorectal cancer and the potential associations with symptom type, socio-demographic characteristics and patient interval. METHODS: Data were collected among incident colorectal cancer patients (n = 577, response rate 64.2 %), who were asked to think back on the time before their diagnosis when completing the questionnaire. The questionnaire comprised a Danish version of the revised Illness Perception Questionnaire (IPQ-R) with questions on 19 symptom attributions. These 19 attribitutions were categorised into five causal groups for statistical analyses. The patient interval (i.e. the time from the patient's first symptom experience to presentation to the healthcare system) was assessed in the same questionnaire. Data on socio-demographic characteristics were obtained by using nationwide registers from Statistics Denmark. RESULTS: Patients who experienced 'blood in stool' as the most important symptom were more likely to attribute this to cancer (PR(ad) 1.94, 95 % CI 1.46-2.58) and benign somatic causes (PR(ad) 1.36, 95 % CI 1.05-1.76), such as haemorrhoids, compared to patients who did not perceive this symptom as the most important. Socio-demographic characteristics were also associated with symptom attribution. Patients with higher educational levels were less likely to attribute their most important symptom to psychological causes (PR(ad) 0.57, 95 % CI 0.34-0.96) than patients with lower educational levels. Patients with rectal cancer attributed their most important symptom to a benign somatic cause more often than patients with colon cancer (PR(ad) 1.34, 95 % CI 1.02-1.77). [corrected]. CONCLUSIONS: Symptom attribution in patients was associated with aspects of socio-demography and with the symptom type perceived by the patient as the most important. No significant associations were found between symptom attributions and patient interval. These results have implications for general practice as symptom attributions may prompt patients to present symptoms in a certain way and thereby influence the general practitioner's assessment of presented symptoms.

High expression of KCa3.1 in patients with clear cell renal carcinoma predicts high metastatic risk and poor survival.

BACKGROUND: Ca2+-activated K+ channels have been implicated in cancer cell growth, metastasis, and tumor angiogenesis. Here we hypothesized that high mRNA and protein expression of the intermediate-conductance Ca2+-activated K+ channel, KCa3.1, is a molecular marker of clear cell Renal Cell Carcinoma (ccRCC) and metastatic potential and survival. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed channel expression by qRT-PCR, immunohistochemistry, and patch-clamp in ccRCC and benign oncocytoma specimens, in primary ccRCC and oncocytoma cell lines, as well as in two ccRCC cell lines (Caki-1 and Caki-2). CcRCC specimens contained 12-fold higher mRNA levels of KCa3.1 than oncocytoma specimens. The large-conductance channel, KCa1.1, was 3-fold more highly expressed in ccRCC than in oncocytoma. KCa3.1 mRNA expression in ccRCC was 2-fold higher than in the healthy cortex of the same kidney. Disease specific survival trended towards reduction in the subgroup of high-KCa3.1-expressing tumors (p<0.08 vs. low-KCa3.1-expressing tumors). Progression-free survival (time to metastasis/recurrence) was reduced significantly in the subgroup of high-KCa3.1-expressing tumors (p<0.02, vs. low-KCa3.1-expressing tumors). Immunohistochemistry revealed high protein expression of KCa3.1 in tumor vessels of ccRCC and oncocytoma and in a subset of ccRCC cells. Oncocytoma cells were devoid of KCa3.1 protein. In a primary ccRCC cell line and Caki-1/2-ccRCC cells, we found KCa3.1-protein as well as TRAM-34-sensitive KCa3.1-currents in a subset of cells. Furthermore, Caki-1/2-ccRCC cells displayed functional Paxilline-sensitive KCa1.1 currents. Neither KCa3.1 nor KCa1.1 were found in a primary oncocytoma cell line. Yet KCa-blockers, like TRAM-34 (KCa3.1) and Paxilline (KCa1.1), had no appreciable effects on Caki-1 proliferation in-vitro. CONCLUSIONS/SIGNIFICANCE: Our study demonstrated expression of KCa3.1 in ccRCC but not in benign oncocytoma. Moreover, high KCa3.1-mRNA expression levels were indicative of low disease specific survival of ccRCC patients, short progression-free survival, and a high metastatic potential. Therefore, KCa3.1 is of prognostic value in ccRCC.