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Abstract Background It has recently been shown that overexpression of the serine protease, matriptase, in transgenic mice causes a dramatically increased frequency of carcinoma formation. Overexpression of HAI-1 and matriptase together changed the frequency of carcinoma formation to normal. This suggests that the ratio of matriptase to HAI-1 influences the malignant progression. The aim of this study has been to determine the ratio of matriptase to HAI-1 mRNA expression in affected and normal tissue from individuals with colorectal cancer adenomas and carcinomas as well as in healthy individuals, in order to determine at which stages a dysregulated ratio of matriptase/HAI-1 mRNA is present during carcinogenesis. Methods Using quantitative RT-PCR, we have determined the mRNA levels for matriptase and HAI-1 in colorectal cancer tissue (n = 9), severe dysplasia (n = 15), mild/moderate dysplasia (n = 21) and in normal tissue from the same individuals. In addition, corresponding tissue was examined from healthy volunteers (n = 10). Matriptase and HAI-1 mRNA levels were normalized to β-actin. Results Matriptase mRNA level was lower in carcinomas compared to normal tissue from healthy individuals (p < 0.01). In accordance with this, the matriptase mRNA level was also lower in adenomas/carcinomas combined as compared to their adjacent normal tissue (p < 0.01). HAI-1 mRNA levels in both normal and affected tissue from individuals with severe dysplasia or carcinomas and in affected tissue with mild/moderate dysplasia were all significantly lower than mRNA levels observed in corresponding tissue from healthy control individuals. HAI-1 mRNA was lower in carcinomas as compared to normal tissue from healthy individuals (p < 0.001). HAI-1 mRNA levels were significantly lower in tissue displaying mild/moderate (p < 0.001) and severe (p < 0.01) dysplasia compared to normal tissue from the same patients. Both adenomas and carcinomas displayed a significantly different matriptase/HAI-1 mRNA ratio than corresponding normal tissue from healthy control individuals (p < 0.05). In addition statistically significant difference (p < 0.001) could be observed between mild/moderate and severe adenomas and their adjacent normal tissue. Conclusion Our results show that dysregulation of the matriptase/HAI-1 mRNA ratio occurs early during carcinogenesis. Future studies are required to clarify whether the dysregulated matriptase/HAI-1 ratio was causing the malignant progression or is a consequence of the same.

Anthropogenic impact has transitioned from threatening already rare species to causing significant declines in once numerous organisms. Long-tailed duck ( Clangula hyemalis ) and velvet scoter ( Melanitta fusca ) were once important quarry sea duck species in NW Europe, but recent declines resulted in their reclassification as vulnerable on the IUCN Red List. We sequenced and assembled genomes for both species and resequenced 15 individuals of each. Using analyses based on site frequency spectra and sequential Markovian coalescence, we found C . hyemalis to show more historical demographic stability, whereas M. fusca was affected particularly by the Last (Weichselian) Glaciation. This likely reflects C . hyemalis breeding continuously across the Arctic, with cycles of glaciation primarily shifting breeding areas south or north without major population declines, whereas the more restricted southern range of M. fusca would lead to significant range contraction during glaciations. Both species showed evidence of declines over the past thousands of years, potentially reflecting anthropogenic pressures with the recent decline indicating an accelerated process. Analysis of runs of homozygosity (ROH) showed low but nontrivial inbreeding, with F ROH from 0.012 to 0.063 in C . hyemalis and ranging from 0 to 0.047 in M. fusca . Lengths of ROH suggested that this was due to ongoing background inbreeding rather than recent declines. Overall, despite demographically important declines, this has not yet led to strong inbreeding and genetic erosion, and the most pressing conservation concern may be the risk of density-dependent (Allee) effects. We recommend monitoring of inbreeding using ROH analysis as a cost-efficient method to track future developments to support effective conservation of these species., Competing Interests: The authors declare no conflicts of interest., (© 2024 The Author(s). Evolutionary Applications published by John Wiley & Sons Ltd.)

Objective: To determine recurrence incidence after partial nephrectomy (PN) for renal cell carcinoma and identify predictors for local recurrence (LR) and metastasis., Material and Methods: We retrospectively evaluated a cohort of 524 patients from the Cancer Registry of Norway, who underwent PN between January 2014 and December 2015 and were followed-up for >6 years. Patient demographics and pathological characteristics were correlated with recurrence and progression-free survival using Kaplan-Meier and Cox regression analyses., Results: Median patient age was 64 years, and the median tumour size was 2.6 cm. A positive surgical margin (PSM) was observed in 11% of the cases, while the LR and metastasis rates were 3.4% and 3.2%, respectively. PSM (hazard ratio [HR], 55.4; 95% confidence interval [CI], 12.55-244.6), tumour number (HR, 45.4; 95% CI, 6.5-316.1) and stage (HR, 33.5; 95% CI, 5.4-205.3) were independent predictors for LR. Undetermined margin status was also a risk factor for LR. Tumour stage (HR, 41.05; 95% CI, 8.52-197.76), tumour necrosis (HR, 1.3; 95% CI, 0.4-4.31) and age (HR, 1.07; 95% CI, 1.01-1.14) were predictors for metastasis., Conclusions: Both local and distant recurrences after PN were rare, and the pT stage was a common predictor. PSM or indeterminate surgical margin and tumour number were LR predictors, while age at surgery and the presence of tumour necrosis predicted metastasis.

Calmodulin (CaM) engages in Ca 2+ -dependent interactions with numerous proteins, including a still incompletely understood physical and functional interaction with the human Na + /H + -exchanger NHE1. Using nuclear magnetic resonance (NMR) spectroscopy, isothermal titration calorimetry, and fibroblasts stably expressing wildtype and mutant NHE1, we discovered multiple accessible states of this functionally important complex existing in different NHE1:CaM stoichiometries and structures. We determined the NMR solution structure of a ternary complex in which CaM links two NHE1 cytosolic tails. In vitro , stoichiometries and affinities could be tuned by variations in NHE1:CaM ratio and calcium ([Ca 2+ ]) and by phosphorylation of S648 in the first CaM-binding α-helix. In cells, Ca 2+ -CaM-induced NHE1 activity was reduced by mimicking S648 phosphorylation and by mutation of the first CaM-binding α-helix, whereas it was unaffected by inhibition of Akt, one of several kinases phosphorylating S648. Our results demonstrate a diversity of NHE1:CaM interaction modes and suggest that CaM may contribute to NHE1 dimerization and thereby augment NHE1 regulation. We propose that a similar structural diversity is of relevance to many other CaM complexes., Competing Interests: LS, AP, EP, MS, KK, JO, BK, SP No competing interests declared, (© 2021, Sjøgaard-Frich et al.)

Dynamic interactions of proteins with lipid membranes are essential regulatory events in biology, but remain rudimentarily understood and particularly overlooked in membrane proteins. The ubiquitously expressed membrane protein Na + /H + -exchanger 1 (NHE1) regulates intracellular pH (pH i ) with dysregulation linked to e.g. cancer and cardiovascular diseases. NHE1 has a long, regulatory cytosolic domain carrying a membrane-proximal region described as a lipid-interacting domain (LID), yet, the LID structure and underlying molecular mechanisms are unknown. Here we decompose these, combining structural and biophysical methods, molecular dynamics simulations, cellular biotinylation- and immunofluorescence analysis and exchanger activity assays. We find that the NHE1-LID is intrinsically disordered and, in presence of membrane mimetics, forms a helical αα-hairpin co-structure with the membrane, anchoring the regulatory domain vis-a-vis the transport domain. This co-structure is fundamental for NHE1 activity, as its disintegration reduced steady-state pH i and the rate of pH i recovery after acid loading. We propose that regulatory lipid-protein co-structures may play equally important roles in other membrane proteins.

Background: Exercise-induced rhabdomyolysis is an increasingly frequent cause of hospitalisation and is much debated in the media. The real incidence of the condition is unknown. We wanted to investigate changes in creatine kinase (CK) levels in healthy students following intensive exercise and to look for a correlation between CK, pain and previous exercise history., Method and Material: Twenty-four healthy students performed a single intensive workout and acted as their own controls with testing before and after the exercise session., Results: All participants displayed an increase in CK levels after the workout, 58 % to above 5 000 IU/l. CK rose from a median of 104 IU/l (72-212) to a median of 6 071 IU/l (2 815-12 275) on day 4, p < 0.001. A negative Spearman's rank correlation was observed between the frequency of strength training prior to the experiment and the CK increase, rho = -0.477 (p = 0.021)., Interpretation: A major increase in CK levels is a normal phenomenon after intensive exercise, and the amount of the increase is related to previous exercise history. Further studies should evaluate whether patients with exercise-induced rhabdomyolysis should receive the same treatment as those with rhabdomyolysis caused by other mechanisms.

Introduction: Walking is endorsed as health enhancing and is the most common type of physical activity among older adults. Accelerometers are superior to self-reports when measuring steps, however, if they are to be used by clinicians the validity is of great importance. The aim of this study was to investigate the criterion validity of Mother and ActiGraph wGT3X-BT in measuring steps by comparing the devices to a hand tally under controlled conditions in healthy participants., Methods: Thirty healthy participants were fitted with a belt containing the sensor of Mother (Motion Cookie) and ActiGraph. Participants walked on a treadmill for two minutes at each of the following speeds; 3.2, 4.8, and 6.4 km/h. The treadmill walking was video recorded and actual steps were subsequently determined by using a hand tally. Wilcoxon's signed ranks test was used to determine whether Mother and ActiGraph measured an identical number of steps compared to the hand tally. Intraclass correlation coefficients were calculated to determine the relationship and Root Mean Square error was calculated to investigate the average error between the devices and the hand tally. Percent differences (PD) were calculated for between-instrument agreement (Mother vs. the hand tally and ActiGraph vs. the hand tally) and PDs below 3% were interpreted as acceptable and clinically irrelevant., Results: Mother and ActiGraph under-counted steps significantly compared to the hand tally at all walking speeds ( p  < 0.001). Mother had a median of total differences of 9.5 steps (IQR = 10) and ActiGraph 59 steps (IQR = 77). Mother had smaller PDs at all speeds especially at 3.2 km/h (2.5% compared to 26.7%). Mother showed excellent ICC values ≥0.88 (0.51-0.96) at all speeds whilst ActiGraph had poor and fair to good ICC values ranging from 0.03 (-0.09-0.21) at a speed of 3.2 km/h to 0.64 (0.16-0.84) at a speed of 6.4 km/h., Conclusion: Mother provides valid measures of steps at walking speeds of 3.2, 4.8, and 6.4 km/h with clinically irrelevant deviations compared to a hand tally while ActiGraph only provides valid measurements at 6.4 km/h based on the 3% criterion. These results have significant potential for valid objective measurements of low walking speeds. However, further research should investigate the validity of Mother in patients at even slower walking speeds and in free-living conditions., Competing Interests: The authors declare there are no competing interests.

Background: Accelerometers can obtain precise measurements of movements during the day. However, the individual activity pattern varies from day-to-day and there is limited evidence on measurement days needed to obtain sufficient reliability. The aim of this study was to examine variability in accelerometer derived data on sedentary behaviour and physical activity at work and in leisure-time during week days among Danish office employees., Methods: We included control participants (n = 135) from the Take a Stand! Intervention; a cluster randomized controlled trial conducted in 19 offices. Sitting time and physical activity were measured using an ActiGraph GT3X+ fixed on the thigh and data were processed using Acti4 software. Variability was examined for sitting time, standing time, steps and time spent in moderate-to-vigorous physical activity (MVPA) per day by multilevel mixed linear regression modelling., Results: Results of this study showed that the number of days needed to obtain a reliability of 80% when measuring sitting time was 4.7 days for work and 5.5 days for leisure time. For physical activity at work, 4.0 days and 4.2 days were required to measure steps and MVPA, respectively. During leisure time, more monitoring time was needed to reliably estimate physical activity (6.8 days for steps and 5.8 days for MVPA)., Conclusions: The number of measurement days needed to reliably estimate activity patterns was greater for leisure time than for work time. The domain specific variability is of great importance to researchers and health promotion workers planning to use objective measures of sedentary behaviour and physical activity., Trial Registration: Clinical trials NCT01996176 .