Pioglitazone does not affect vascular or inflammatory responses after endotoxemia in humans.

PPARgamma agonists have been proposed to exert more than metabolic benefits, particularly by anti-inflammatory mechanisms. We hypothesized that pioglitazone might modulate inflammatory and vascular responses to lipopolysaccharide (LPS). In a placebo-controlled parallel-group study in 18 healthy male subjects, the E. coli endotoxin model of inflammation (20 IU/kg i. v.) was employed to test the effect of 60 mg pioglitazone over nine days on inflammatory cytokines. Macrovascular function and microvascular blood flow were assessed by brachial artery ultrasound and retinal blood flow parameters, respectively. Pioglitazone increased brachial artery diameter by 5.6% but had no effect on other outcome parameters under resting conditions. LPS increased cytokine levels to peak concentrations of 91.3+/-22.5 ng/ml (IL-6), 261.4+/-60.0 ng/ml (TNFalpha), and 524.5+/-15.3 ng/ml (VCAM-1). The endotoxin caused microvascular vasodilation and increased retinal white blood cell flux, while baseline brachial artery diameter remained unchanged. Pioglitazone had no effect on inflammatory cytokine or adhesion molecule release but mitigated LPS-induced hypotension (p<0.05). Neither brachial artery function nor microvascular blood flow was altered by pioglitazone. In conclusion, acute immune reactions to LPS are not affected by pioglitazone, which exerts subtle vascular effects alone and during endotoxemia. The anti-inflammatory properties of short-term pioglitazone to endotoxins in healthy subjects are therefore limited.     

Function and quality of kidneys after cold storage, machine perfusion, or retrograde oxygen persufflation: Results from a porcine autotransplantation model

Objectives

Especially for preservation of marginal donor organs, machine perfusion (MP) and retrograde oxygen persufflation (ROP) are alternatives to cold storage (CS). Using a porcine kidney autotransplantation model we compared metabolic and morphologic effects of CS, ROP, and MP on kidneys exposed to warm ischemia.

Methods

Kidneys of 21 pigs were exposed in situ to warm ischemia for 60 min. The kidneys were randomly allocated to three experimental groups, each receiving a 4-h treatment of either cold storage, machine perfusion, or retrograde oxygen persufflation. Tissue samples were examined for malondialdehyde and histological changes. Daily blood samples were examined for creatinine levels.

Results

Seven days after transplantation, the plasma creatinine levels in the CS and MP groups were still significantly elevated above the baseline values. In the ROP group, all animals exhibited nearly normal creatinine levels. Malondialdehyde, an indicator of lipidperoxidation, was dramatically increased in the machine perfused kidneys on day 7, whereas the malondialdehyde levels in the other two groups were near normal values. The MP kidneys exhibited the most striking histological changes.

Conclusion

Though MP has been well introduced in organ transplantation, in our opinion, it must still be optimized and standardized. It is necessary to clarify questions such as whether there is a need for oxygenation during perfusion, the length of perfusion, the impact of pressure, and the effects of additional scavengers. The results of the present study suggest the reconsideration of the ROP-technique for the preservation of predamaged donor grafts especially of NHBD and further studies, comparing MP and ROP upon long term preservation are strongly encouraged.     

Ringing or colour-banding does not increase predation mortality in redshanksTringa totanus

The use of metal and colour‐rings or bands as a means of measuring survival, movements and behaviour in birds is universal and fundamental to testing ecological and evolutionary theories. The practice rests on the largely untested assumption that the rings do not affect survival. However this assumption may not hold for several reasons, for example because the ‘oddity effect’ predicts predators select prey that appear different to their neighbours in order to avoid the ‘confusion effect’. We compared the foraging behaviour and the death rates of redshanks Tringa totanus conspicuously marked with six colour rings and one metal ring each to unmarked birds in a study system, where routinely up to 50% of the total population are killed by avian predators during a winter. If avian predators selectively target and/or have a higher capture success of ringed birds then we would predict the proportion of colour‐ringed birds in the population to decline through a winter. The proportion of colour‐ringed birds in the population did not change over the course of three separate winters, and in one winter the ratio of marked:unmarked birds found killed by sparrowhawks Accipiter nisus was the same as the ratio of marked birds alive in the population. In the year with largest sample size, power was sufficient to detect a greater than 2.2% difference in predation rate between ringed and unringed groups. The average kill rate difference between ringed and unringed birds across the three winters was less than 1% (0.73±2.2%) suggesting that even if there were differences in predation rate that were not detected because of low statistical power they were extremely small. There were no differences in any foraging measures comparing ringed and unringed birds, suggesting that the rings did not affect the ability of birds to meet their daily energy budgets. The results showed that colour‐ringed birds were not preferentially targeted or killed by avian predators, and suggest that the presence of a metal and even several large colour‐rings is unlikely to affect behaviour and predation mortality even under extreme selection.     

Dystroglycan does not contribute significantly to kidney development or function, in health or after injury

Dystroglycan (DG or DAG1) is considered a critical link between the basement membrane and the cytoskeleton in multiple tissues. DG consists of two subunits, an extracellular α-subunit that binds laminin and other basement membrane components, and a transmembrane β-subunit. DG-null mouse embryos die during early embryogenesis because DG is required for Reichert's membrane formation. DG also forms an integral part of the dystrophin-glycoprotein complex in muscle. Although no human DG mutations have been reported, multiple forms of muscular dystrophy have been linked to DG glycosylation defects, and targeted deletion of muscle DG causes muscular dystrophy in mice. Moreover, DG is widely distributed in endothelial and epithelial cells, including those in the kidney. There has therefore been significant interest in DG's role in the kidney, especially in podocytes. Previous reports suggested that DG's disturbance in podocytes might cause glomerular filtration barrier abnormalities. To fully understand DG's contribution to nephrogenesis and kidney function, we used a conditional DG allele and a variety of Cre mice to systematically delete DG from podocytes, ureteric bud, metanephric mesenchyme, and then from the whole kidney. Surprisingly, none of these conditional deletions resulted in significant morphological or functional abnormalities in the kidney. Furthermore, DG-deficient podocytes did not show increased susceptibility to injury, and DG-deficient kidneys did not show delayed recovery. Integrins are therefore likely the primary extracellular matrix receptors in renal epithelia.     

Roadside connectivity does not increase reptile abundance or richness in a fragmented mallee landscape

The effect of isolation and the importance of dispersal in establishing and maintaining populations in fragments of remnant habitat remain poorly understood. Nevertheless, environmental connectivity is likely to be important for ensuring the long‐term preservation of biodiversity in extensively cleared landscapes. In this study, we compared reptile communities in large conservation parks with those in small woodland remnants 6.5–12 km from the parks, on the Eyre Peninsula, South Australia, Australia. We assessed the impact of fragmentation on the abundance, richness and habitat preferences of reptiles, and examined whether connection to linear roadside vegetation altered reptile communities in small woodland remnants. Of the 31 reptile species, 12 were restricted to conservation parks and six to habitat fragments in farmland. There was a substantial reduction in reptile species richness and abundance in farmland fragments. Direct connection of remnant vegetation to roadside corridors did not affect abundance of common species in the farmland fragments, although species richness was lower in isolated remnants in one of our two study regions. The habitat preference of the scincid lizard Menetia greyii differed between farmland fragments, where they were regularly found on dunes and roadsides, and conservation parks, where they were rare and not detected on dunes. We suggest that habitat fragmentation may have altered interspecific interactions, enabling an expansion of habitat use in the farming landscape. Significantly lower abundance of four common species in farmland settings compared with reserves indicated that existing corridors and small fragments provide inadequate connectivity over larger distances. To counter this effect, large reserves may need to be less than 10 km apart.     

Sympathetic activation by the cold pressor test does not increase the muscle force generation capacity

A positive inotropic action by the sympathetic nervous system on skeletal muscles has been observed and investigated in animal and in vitro studies. This action provided a theoretical basis for the putative ergogenic action of catecholamines and adrenergic agonists, although there is no clear evidence of this effect in humans. The aim of this study was to investigate the occurrence of inotropic effects associated to physiological sympathetic activation in healthy subjects. The muscle force capacity was investigated in the tibialis anterior (n = 9 subjects) and in the soleus (n = 9) muscles electrically stimulated with single pulses and double pulses with variable interspike interval (4-1,000 ms) and short pulse trains (frequency: 5-14 Hz) before, during, and after sympathetic activation by the cold pressor test (CPT). CPT significantly decreased by 10.4 ± 7.2 and 10.6 ± 4.4% the force produced by single and double pulse stimulation, respectively, and produced smaller decreases in the force obtained by train stimulation in the tibialis anterior, while no significant changes were observed in either type of contraction in the soleus muscle. CPT failed to induce any increase in the force capacity of the investigated muscles. The prevalent decrease in force evidenced in this study supports the concept that the weakening sympathetic action on type I fiber, already shown to occur in humans, prevails over the putative potentiating action.     

Theophylline does not increase maximal tetanic force or diaphragm endurance in vitro

These experiments tested the capacity of theophylline to improve diaphragm strength (maximal force development) and endurance (maintenance of force output during repeated contractions). Rodent diaphragm strips were mounted at optimal length in oxygenated Krebs-Ringer solution (37 degrees C, pH 7.37). Direct stimuli used supramaximal current density, 0.2-ms pulses, and 250-ms tetanic trains. Theophylline (500 mg/ml) increased force development at low stimulation frequencies but did not increase maximal force [25.7 +/- 0.5 for theophylline vs. 26.0 +/- 0.4 (SE) N/cm2 for control (n = 34)]. During repeated submaximal (25-36 Hz) tetanic contractions, theophylline did not affect endurance. During repeated maximal (160 Hz) tetanic contractions theophylline reduced endurance, accelerating the fall of developed force. Theophylline also inhibited recovery of force after endurance trials ended. We conclude that theophylline does not increase maximal tetanic force and can reduce diaphragm endurance in vitro.     

Osteocalcin does not influence acute or chronic inflammation in human vascular cells

Some human observational studies have suggested an anti-inflammatory role of osteocalcin (OCN). An inflammatory protocol using interferon-γ and tumor necrosis factor-α (10 ng/ml) was employed to examine the acute (24 hr) and chronic (144 hr) effects of uncarboxylated OCN (ucOCN) in commercial, primary, subcultured human aortic endothelial cells (HAEC), and human smooth muscle cells (HASMCs). The inflammatory protocol increased phosphorylation of intracellular signaling proteins (CREB, JNK, p38, ERK, AKT, STAT3, STAT5) and increased secretion of adhesion markers (vascular cell adhesion molecule-1, intracellular adhesion molecule-1, monocyte chemoattractant protein-1) and proinflammatory cytokines (interleukin-6 [IL-6], IL-8). After acute inflammation, there were no additive or reductive effects of ucOCN in either cell type. Following chronic inflammation, ucOCN did not affect cell responses, nor did it appear to have any pro- or anti-inflammatory effects when administered acutely or chronically on its own in either cell type. Additionally, ucOCN did not affect lipopolysaccharide (LPS)-induced acute inflammation in HAECs or HASMCs. The findings of this study do not support a causal role for OCN within the models of vascular inflammation chosen. Further confirmatory studies are warranted.     

Effects of short-term hypothermic perfusion and cold storage on function of the isolated-perfused dog kidney

The isolated-perfused dog kidney was used as a model to measure the effects of short-term hypothermic preservation on renal function and metabolism. Kidneys were cold-stored in Collins' solution, hypotonic citrate, or phosphate-buffered sucrose for 4 and 24 hr, or were continuously perfused for 4 and 24 hr with a synthetic perfusate. Following preservation kidneys were perfused with an albumin-containing perfusate at 37 degrees C for 60 min for determination of renal function. The results indicate that many of the effects of short-term preservation on renal function in dog kidneys are similar to results reported for rat and rabbit kidneys. Cold storage for 4 hr resulted in a large decrease in GFR (57%), but only a small decrease in Na reabsorption (from 97 to 87%). Cold storage for 24 hr caused a further decline in renal function (GFR = 95% decrease, Na reabsorption = 49-64%). Results were similar for all cold storage solutions tested. Perfusion for 4 hr was less damaging to renal function than cold storage. The GFR decreased only 14% and urine formation and Na reabsorption were practically normal. After 24 hr of hypothermic perfusion, the GFR was reduced by 79%, urine flow was normal, and Na reabsorption was 78%. There were no obvious biochemical correlates (adenine nucleotides, tissue edema, or electrolyte concentration) with the loss of renal function during short-term preservation. The results suggest that the isolated-perfused dog kidney can be used to test the effects of preservation on renal function, and yields results similar to those obtained using small animal models.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increasing pulse wave velocity in a realistic cardiovascular model does not increase pulse pressure with age

The mechanism of the well-documented increase in aortic pulse pressure (PP) with age is disputed. Investigators assuming a classical windkessel model believe that increases in PP arise from decreases in total arterial compliance (C(tot)) and increases in total peripheral resistance (R(tot)) with age. Investigators assuming a more sophisticated pulse transmission model believe PP rises because increases in pulse wave velocity (c(ph)) make the reflected pressure wave arrive earlier, augmenting systolic pressure. It has recently been shown, however, that increases in c(ph) do not have a commensurate effect on the timing of the reflected wave. We therefore used a validated, large-scale, human arterial system model that includes realistic pulse wave transmission to determine whether increases in c(ph) cause increased PP with age. First, we made the realistic arterial system model age dependent by altering cardiac output (CO), R(tot), C(tot), and c(ph) to mimic the reported changes in these parameters from age 30 to 70. Then, c(ph) was theoretically maintained constant, while C(tot), R(tot), and CO were altered. The predicted increase in PP with age was similar to the observed increase in PP. In a complementary approach, C(tot), R(tot), and CO were theoretically maintained constant, and c(ph) was increased. The predicted increase in PP was negligible. We found that increases in c(ph) have a limited effect on the timing of the reflected wave but cause the system to degenerate into a windkessel. Changes in PP can therefore be attributed to a decrease in C(tot).     

Desaturation function does not decline after menopause in human females.

Aging appears to decrease delta6-desaturase activity in males, but in females it is uncertain. delta6- and delta5-desaturase functions were investigated in pre- and post-menopausal women who were normoglycemic or had type 2 diabetes (2 x 2 factorial, n = 37). Subjects were compared for indicators of diabetic control, estrogen levels, fatty acid profiles and indices of delta6- and delta5-desaturase activity. Diet intakes that were compared to determine whether results were a function of dietary factors known to influence desaturase activity revealed no differences (P>0.05). Post-menopausal women with type 2 diabetes had more 18:2 n6 in serum phospholipids (P<0.05) than did the pre- and post-menopausal control subjects. Fatty acid ratios of 18:3 n6/18:2 n6 indicated greater delta6-desaturase activity for women with type 2 diabetes, but differences were not found between pre- and post-menopausal groups. Significant correlation (P < 0.05) indicates an association between diabetic status and desaturase function, but function did not appear to be affected by menopausal status. In contrast to reports using male subjects, we found no evidence that desaturase function decreased in aging females, as reported for males, or increased as hypothesized in this study.     

Creatine supplementation during college football training does not increase the incidence of cramping or injury

The purpose of this study was to examine the effects of creatine supplementation on the incidence of injury observed during 3-years of NCAA Division IA college football training and competition. In an open label manner, athletes participating in the 1998–2000 football seasons elected to take creatine or non-creatine containing supplements following workouts/practices. Subjects who decided to take creatine were administered 15.75 g of creatine for 5 days followed by ingesting an average of 5 g/day thereafter administered in 5–10 g doses. Creatine intake was monitored and recorded by research assistants throughout the study and ranged between 34–56% of players during the course of the study. Subjects practiced or played in environmental conditions ranging from 8–40°C (mean 24.7 ± 9°C) and 19–98% relative humidity (49.3 ± 17%). Injuries treated by the athletic training staff were recorded and categorized as cramping, heat/dehydration, muscle tightness, muscle strains/pulls, non-contact joint injuries, contact injuries, and illness. The number of missed practices due to injury/illness was also recorded. Data are presented as the total number of treated injuries for creatine users/total injuries observed and percentage occurrence rate of injuries for creatine users for all seasons. The incidence of cramping (37/96, 39%), heat/dehydration (8/28, 36%), muscle tightness (18/42, 43%), muscle pulls/strains (25/51, 49%), non-contact joint injuries (44/132, 33%), contact injuries (39/104, 44%), illness (12/27, 44%), number of missed practices due to injury (19/41, 46%), players lost for the season (3/8, 38%), and total injuries/missed practices (205/529, 39%) were generally lower or proportional to the creatine use rate among players. Creatine supplementation does not appear to increase the incidence of injury or cramping in Division IA college football players.     

Single dose of synthetic microRNA-199a or microRNA-149 mimic does not improve cardiac function in a murine model of myocardial infarction

Molecular and Cellular Biochemistry - Intramyocardial injection of synthetic microRNAs (miRs) has recently been reported to be beneficial after myocardial infarction (MI). We conducted a randomized...     

Nitrate tolerance does not increase production of peroxynitrite in the heart

Clinical studies have suggested that long-term nitrate treatment does not improve and may even worsen cardiovascular mortality, and the possible role of nitrate tolerance has been suspected. Nitrate tolerance has been recently shown to increase vascular superoxide and peroxynitrite production leading to vascular dysfunction. Nevertheless, nitrates exert direct cardiac effects independent from their vascular actions. Therefore, we investigated whether in vivo nitroglycerin treatment leading to vascular nitrate tolerance increases cardiac formation of nitric oxide (NO), reactive oxygen species, and peroxynitrite, thereby leading to cardiac dysfunction. Nitrate tolerance increased bioavailability of NO in the heart without increasing formation of reactive oxygen species. Despite elevated myocardial NO, neither cardiac markers of peroxynitrite formation nor cardiac mechanical function were affected by nitroglycerin treatment. However, serum free nitrotyrosine, a marker for systemic peroxynitrite formation, was significantly elevated in nitroglycerin-treated animals. This is the first demonstration that, although the systemic effects of nitroglycerin may be deleterious due to enhancement of extracardiac peroxynitrite formation, nitroglycerin does not result in oxidative damage in the heart.     

Targeted Sprouty1 overexpression in cardiac myocytes does not alter myocardial remodeling or function

The mitogen activated protein kinase (MAPK) signaling pathway regulates multiple events leading to heart failure including ventricular remodeling, contractility, hypertrophy, apoptosis, and fibrosis. The regulation of conserved intrinsic inhibitors of this pathway is poorly understood. We recently identified an up-regulation of Sprouty1 (Spry1) in a targeted approach for novel inhibitors of the MAPK signaling pathway in failing human hearts following reverse remodeling. The goal of this study was to test the hypothesis that up-regulated expression of Spry1 in cardiac myocytes would be sufficient to inhibit ERK1/2 activation and tissue remodeling. We established a murine model with up-regulated Spry1 expression in cardiac myocytes using the alpha-myosin heavy chain promoter (α-MHC). Heart weight and cardiac myocyte morphology were unchanged in adult male α-MHC–Spry1 mice compared to control mice. Ventricular function of α-MHC–Spry1 mice was unaltered at 8 weeks or 1 year of age. These findings were consistent with the lack of an effect of Spry1 on ERK1/2 activity. In summary, targeted up-regulation of Spry1 in cardiac myocytes is not sufficient to alter cell or tissue remodeling consistent with the lack of an effect on ERK1/2 activity.     

Injected mRNA does not increase protein synthesis in unfertilized, fertilized, or ammonia-activated sea urchin eggs

We have investigated whether the rate of protein synthesis in unfertilized and fertilization-activated sea urchin eggs is limited by the availability of mRNA by injecting eggs, zygotes, and ammonia-activated eggs with globin mRNA. Message-injected and buffer-injected cells were labeled with radioactive amino acids and the proteins separated on a polyacrylamide gel. The relative amounts of newly synthesized globin and endogenous proteins were obtained by scanning the gel fluorograph. Globin mRNA is translated poorly in Strongylocentrotus droebachiensis eggs and does not significantly increase or decrease endogenous protein synthesis. In zygotes and ammonia-activated eggs, however, globin mRNA is translated well and appears to compete with endogenous mRNAs for the limiting component of the translational machinery as it is released. Our results are consistent with the hypothesis that either ribosomes or recruitment factors are gradually activated after fertilization or ammonia treatment, that such components are the rate-limiting factor, and that they impart the typical sigmoidal increase in protein synthesis rate observed in fertilized eggs before the first cleavage.     

Red trap colour of the carnivorous plant Drosera rotundifolia does not serve a prey attraction or camouflage function

The traps of many carnivorous plants are red in colour. This has been widely hypothesized to serve a prey attraction function; colour has also been hypothesized to function as camouflage, preventing prey avoidance. We tested these two hypotheses in situ for the carnivorous plant Drosera rotundifolia. We conducted three separate studies: (i) prey attraction to artificial traps to isolate the influence of colour; (ii) prey attraction to artificial traps on artificial backgrounds to control the degree of contrast and (iii) observation of prey capture by D. rotundifolia to determine the effects of colour on prey capture. Prey were not attracted to green traps and were deterred from red traps. There was no evidence that camouflaged traps caught more prey. For D. rotundifolia, there was a relationship between trap colour and prey capture. However, trap colour may be confounded with other leaf traits. Thus, we conclude that for D. rotundifolia, red trap colour does not serve a prey attraction or camouflage function.