Cytoskeleton stiffness regulates cellular senescence and innate immune response in Hutchinson–Gilford Progeria Syndrome

Hutchinson–Gilford progeria syndrome (HGPS) is caused by the accumulation of mutant prelamin A (progerin) in the nuclear lamina, resulting in increased nuclear stiffness and abnormal nuclear architecture. Nuclear mechanics are tightly coupled to cytoskeletal mechanics via lamin A/C. However, the role of cytoskeletal/nuclear mechanical properties in mediating cellular senescence and the relationship between cytoskeletal stiffness, nuclear abnormalities, and senescent phenotypes remain largely unknown. Here, using muscle‐derived mesenchymal stromal/stem cells (MSCs) from the Zmpste24^?/? (Z24^?/?) mouse (a model for HGPS) and human HGPS fibroblasts, we investigated the mechanical mechanism of progerin‐induced cellular senescence, involving the role and interaction of mechanical sensors RhoA and Sun1/2 in regulating F‐actin cytoskeleton stiffness, nuclear blebbing, micronuclei formation, and the innate immune response. We observed that increased cytoskeletal stiffness and RhoA activation in progeria cells were directly coupled with increased nuclear blebbing, Sun2 expression, and micronuclei‐induced cGAS‐Sting activation, part of the innate immune response. Expression of constitutively active RhoA promoted, while the inhibition of RhoA/ROCK reduced cytoskeletal stiffness, Sun2 expression, the innate immune response, and cellular senescence. Silencing of Sun2 expression by siRNA also repressed RhoA activation, cytoskeletal stiffness and cellular senescence. Treatment of Zmpste24^?/? mice with a RhoA inhibitor repressed cellular senescence and improved muscle regeneration. These results reveal novel mechanical roles and correlation of cytoskeletal/nuclear stiffness, RhoA, Sun2, and the innate immune response in promoting aging and cellular senescence in HGPS progeria.     

The effect of body weight on the degree of blood velocity profile skewness in the aortic annulus in domestic pigs

We investigated the blood velocity profile in the aortic annulus (AA) in two groups of domestic pigs using epicardial Doppler echocardiography. The velocity profile skewness in terms of max/mean TVI (the ratio of maximal to cross-sectional mean time-velocity integral along the diameter) was 1.107 +/- 0.01 in the small pigs (n = 10; body weight: 24.6 +/- 0.8 kg) and 1.216 +/- 0.026 in the large pigs (n = 8; body weight: 50.6 +/- 2.5 kg) (P = 0.002). The velocity profile in the AA is more skewed in large animals than in small animals and the skewness in the larger animals is similar to that in normal adult humans. This study shows the importance of choosing animals of sufficient size if flow method investigations are to be performed. This is particularly important for ultrasound Doppler investigations based on a limited sample of velocities across the flow channel.     

Postnatal deficiency of essential fatty acids in mice results in resistance to diet-induced obesity and low plasma insulin during adulthood

Our objective was to investigate the long-term metabolic effects of postnatal essential fatty acid deficiency (EFAD). Mouse dams were fed an EFAD diet or an isoenergetic control diet 4 days before delivery and throughout lactation. The pups were weaned to standard diet (STD) and were later subdivided into two groups: receiving high fat diet (HFD) or STD. Body composition, energy expenditure, food intake and leptin levels were analyzed in adult offspring. Blood glucose and plasma insulin concentrations were measured before and during a glucose tolerance test. EFAD offspring fed STD were leaner with lower plasma leptin and insulin concentrations compared to controls. EFAD offspring fed HFD were resistant to diet-induced obesity, had higher energy expenditure and lower levels of plasma leptin and insulin compared to controls. These results indicate that the fatty acid composition during lactation is important for body composition and glucose tolerance in the adult offspring.     

Targeted and global pharmacometabolomics in everolimus-based immunosuppression: association of co-medication and lysophosphatidylcholines with dose requirement

Introduction

The immunosuppressive therapy with everolimus (ERL) after heart transplantation is characterized by a narrow therapeutic window and a substantial variability in dose requirement. Factors explaining this variability are largely unknown.

Objectives

Our aim was to evaluate factors affecting ERL metabolism and to identify novel metabolites associated with the individual ERL dose requirement to elucidate mechanisms underlying ERL dose response variability.

Method

We used liquid chromatography coupled with mass spectrometry for quantification of ERL metabolites in 41 heart transplant patients and evaluated the effect of clinical and genetic factors on ERL pharmacokinetics. Non-targeted plasma metabolic profiling by ultra-performance liquid chromatography and high resolution quadrupole-time-of-flight mass spectrometry was used to identify novel metabolites associated with ERL dose requirement.

Results

The determination of ERL metabolites revealed differences in metabolite patterns that were independent from clinical or genetic factors. Whereas higher ERL dose requirement was associated with co-administration of sodium-mycophenolic acid and the CYP3A5 expressor genotype, lower dose was required for patients receiving vitamin K antagonists. Global metabolic profiling revealed several novel metabolites associated with ERL dose requirement. One of them was identified as lysophosphatidylcholine (lysoPC) (16:0/0:0). Subsequent targeted analysis revealed that high levels of several lysoPCs were significantly associated with higher ERL dose requirement.

Conclusion

For the first time, this study describes distinct ERL metabolite patterns in heart transplant patients and detected potentially new drug–drug interactions. The global metabolic profiling facilitated the discovery of novel metabolites associated with ERL dose requirement that might represent new clinically valuable biomarkers to guide ERL therapy.

     

The association of sleepiness,insomnia, sleep disturbance and pain: a study amongst shiftworking nurses

Sleep and Biological Rhythms - Sleep problems are commonly associated with chronic pain. It is not known whether pain is more related to a particular type of sleep problem or to more composite...     

Changes in lysosome populations in the rat kidney cortex induced by experimental proteinuria

1. Experimental proteinuria (262.9 mg protein/24 hr urine) was induced in rats by repeated intraperitoneal injections of BSA. 2. Hypertrophy of the kidney cortex was significant 8 days after the start of the BSA injections, and the activities of lysosomal enzymes in kidney cortex and urine were significantly higher in proteinuric compared to nonproteinuric rats. 3. Lysosome populations in the kidney cortex were examined by rate sedimentation of the homogenate and by rate zonal and isopycnic centrifugation of the lysosome-rich ML fraction. 4. The activity of lysosomal enzymes in the kidney cortex increased slightly, essentially in the large, fragile lysosomes mainly recovered from the proximal tubule. 5. Proteinuria induced a shift/reduction in the density of small lysosomes from 1.235 and 1.20 g/ml to 1.225 and 1.185 g/ml, respectively. 6. Proteinuria induced a new population of small lysosomes (density 1.185 g/ml) enriched in cathepsin D.     

Nitrogen Removal by Riparian Buffers along a European Climatic Gradient: Patterns and Factors of Variation

We evaluated nitrogen (N) removal efficiency by riparian buffers at 14 sites scattered throughout seven European countries subject to a wide range of climatic conditions. The sites also had a wide range of nitrate inputs, soil characteristics, and vegetation types. Dissolved forms of N in groundwater and associated hydrological parameters were measured at all sites; these data were used to calculate nitrate removal by the riparian buffers. Nitrate removal rates (expressed as the difference between the input and output nitrate concentration in relation to the width of the riparian zone) were mainly positive, ranging from 5% m⁻¹ to 30% m⁻¹, except for a few sites where the values were close to zero. Average N removal rates were similar for herbaceous (4.43% m⁻¹) and forested (4.21% m⁻¹) sites. Nitrogen removal efficiency was not affected by climatic variation between sites, and no significant seasonal pattern was detected. When nitrate inputs were low, a very large range of nitrate removal efficiencies was found both in the forested and in the nonforested sites. However, sites receiving nitrate inputs above 5 mg N L⁻¹ showed an exponential negative decay of nitrate removal efficiency (nitrate removal efficiency = 33.6 e^{−0.11 NO3input}, r ² = 0.33, P < 0.001). Hydraulic gradient was also negatively related to nitrate removal (r = −0.27, P < 0.05) at these sites. On the basis of this intersite comparison, we conclude that the removal of nitrate by biological mechanisms (for example, denitrification, plant uptake) in the riparian areas is related more closely to nitrate load and hydraulic gradient than to climatic parameters. Received 15 August 2001; accepted 2 May 2002.