Pulse mTOR inhibitor treatment effectively controls cyst growth but leads to severe parenchymal and glomerular hypertrophy in rat polycystic kidney disease

The efficacy of mammalian target of rapamycin (mTOR) inhibitors is currently tested in patients affected by autosomal dominant polycystic kidney disease. Treatment with mTOR inhibitors has been associated with numerous side effects. However, the renal-specific effect of mTOR inhibitor treatment cessation in polycystic kidney disease is currently unknown. Therefore, we compared pulse and continuous everolimus treatment in Han:SPRD rats. Four-week-old male heterozygous polycystic and wild-type rats were administered everolimus or vehicle by gavage feeding for 5 wk, followed by 7 wk without treatment, or continuously for 12 wk. Cessation of everolimus did not result in the appearance of renal cysts up to 7 wk postwithdrawal despite the reemergence of S6 kinase activity coupled with an overall increase in cell proliferation. Pulse everolimus treatment resulted in striking noncystic renal parenchymal enlargement and glomerular hypertrophy that was not associated with compromised kidney function. Both treatment regimens ameliorated kidney function, preserved the glomerular-tubular connection, and reduced proteinuria. Pulse treatment at an early age delays cyst development but leads to striking glomerular and parenchymal hypertrophy. Our data might have an impact when long-term treatment using mTOR inhibitors in patients with autosomal dominant polycystic kidney disease is being considered.

Human bronchial epithelium controls TH2 responses by TH1-induced, nitric oxide-mediated STAT5 dephosphorylation: implications for the pathogenesis of asthma

Increased levels of NO in exhaled air in association with increased NO synthetase (NOS)2 expression in bronchial epithelial are hallmark features of asthma. It has been suggested that NO contributes to asthma pathogenesis by selective down-regulation of TH1 responses. We demonstrate, however, that NO can reversibly limit in vitro expansion of both human TH1 and TH2 CD4+ T cells. Mechanistically, NO induces cGMP-mediated reversible STAT5 dephosphorylation and therefore interferes with the IL-2R activation cascade. Human bronchial epithelial cells (HBEC) up-regulate NOS2 after stimulation with IFN-gamma secreted by TH1 CD4+ T cells and release NO, which inhibits both TH1 and TH2 cell proliferation. This reversible T cell growth arrest depends on NO because T cell proliferation is completely restored after in vitro blocking of NOS2 on HBEC. HBEC thus drive the effector end of a TH1-controlled feedback loop, which protects airway mucosal tissues at the potential lesional site in asthma from overwhelming CD4+ TH2 (and potentially TH1) responses following allergen exposure. Variations in the efficiency of this feedback loop provides a plausible mechanism to explain why only a subset of atopics sensitized to ubiquitous aeroallergens progress to expression of clinically relevant levels of airways inflammation.

Quality controls for two heel bone ultrasounds used in the Swiss Evaluation of the Methods of Measurement of Osteoporotic Fracture Risk Study

Because of the important morbidity and mortality associated with osteoporosis, it is essential to detect subjects at risk by screening methods, such as bone quantitative ultrasounds (QUSs). Several studies showed that QUS could predict fractures. None, however, compared prospectively different QUS devices, and few data of quality controls (QCs) have been published. The Swiss Evaluation of the Methods of Measurement of Osteoporotic Fracture Risk is a prospective multicenter study that compared three QUSs for the assessment of hip fracture risk in a population of 7609 women age >/=70 yr. Because the inclusion phase lasted 20 mo, and because 10 centers participated in this study, QC became a major issue. We therefore developed a QC procedure to assess the stability and precision of the devices, and for their cross-calibration. Our study focuses on the two heel QUSs. The water bath system (Achilles+) had a higher precision than the dry system (Sahara). The QC results were highly dependent on temperature. QUS stability was acceptable, but Sahara must be calibrated regularly. A sufficient homogeneity among all the Sahara devices could be demonstrated, whereas significant differences were found among the Achilles+ devices. For speed of sound, 52% of the differences among the Achilles+ was explained by the water s temperature. However, for broadband ultrasound attenuation, a maximal difference of 23% persisted after adjustment for temperature. Because such differences could influence measurements in vivo, it is crucial to develop standardized phantoms to be used in prospective multicenter studies.

Effect of oral calcium loading on intact PTH and calcitriol in idiopathic renal calcium stone formers and healthy controls

The calciuric response after an oral calcium load (1000 mg elemental calcium together with a standard breakfast) was studied in 13 healthy male controls and 21 recurrent idiopathic renal calcium stone formers, 12 with hypercalciuria (UCa x V > 7.50 mmol/24 h) and nine with normocalciuria. In controls, serum 1,25(OH)2 vitamin D3 (calcitriol) remained unchanged 6 h after oral calcium load (50.6 +/- 5.1 versus 50.9 +/- 5.0 pg/ml), whereas it tended to increase in hypercalciuric (from 53.6 +/- 3.2 to 60.6 +/- 5.4 pg/ml, P = 0.182) and fell in normocalciuric stone formers (from 45.9 +/- 2.6 to 38.1 +/- 3.3 pg/ml, P = 0.011). The total amount of urinary calcium excreted after OCL was 2.50 +/- 0.20 mmol in controls, 2.27 +/- 0.27 mmol in normocalciuric and 3.62 +/- 0.32 mmol in hypercalciuric stone formers (P = 0.005 versus controls and normocalciuric stone formers respectively); it positively correlated with serum calcitriol 6 h after calcium load (r = 0.392, P = 0.024). Maximum increase in urinary calcium excretion rate, delta Ca-Emax, was inversely related to intact PTH levels in the first 4 h after calcium load, i.e. more pronounced PTH suppression predicted a steeper increase in urinary calcium excretion rate. Twenty-four-hour urine calcium excretion rate was inversely related to the ratio of delta calcitriol/deltaPTHmax after calcium load (r = -0.653, P = 0.0001), indicating that an abnormally up-regulated synthesis of calcitriol and consecutive relative PTH suppression induce hypercalciuria.(ABSTRACT TRUNCATED AT 250 WORDS)

Continuous monitoring of bovine spongiform encephalopathy rapid test performance by weak positive tissue controls and quality control charts

Bovine spongiform encephalopathy (BSE) rapid tests and routine BSE-testing laboratories underlie strict regulations for approval. Due to the lack of BSE-positive control samples, however, full assay validation at the level of individual test runs and continuous monitoring of test performance on-site is difficult. Most rapid tests use synthetic prion protein peptides, but it is not known to which extend they reflect the assay performance on field samples, and whether they are sufficient to indicate on-site assay quality problems. To address this question we compared the test scores of the provided kit peptide controls to those of standardized weak BSE-positive tissue samples in individual test runs as well as continuously over time by quality control charts in two widely used BSE rapid tests. Our results reveal only a weak correlation between the weak positive tissue control and the peptide control scores. We identified kit-lot related shifts in the assay performances that were not reflected by the peptide control scores. Vice versa, not all shifts indicated by the peptide control scores indeed reflected a shift in the assay performance. In conclusion these data highlight that the use of the kit peptide controls for continuous quality control purposes may result in unjustified rejection or acceptance of test runs. However, standardized weak positive tissue controls in combination with Shewhart-CUSUM control charts appear to be reliable in continuously monitoring assay performance on-site to identify undesired deviations.

A bHLH Complex Controls Embryonic Vascular Tissue Establishment and Indeterminate Growth in Arabidopsis

Plants have a remarkable potential for sustained (indeterminate) postembryonic growth. Following their specification in the early embryo, tissue-specific precursor cells first establish tissues and later maintain them postembryonically. The mechanisms underlying these processes are largely unknown. Here we define local control of oriented, periclinal cell division as the mechanism underlying both the establishment and maintenance of vascular tissue. We identify an auxin-regulated basic helix-loop-helix (bHLH) transcription factor dimer as a critical regulator of vascular development. Due to a loss of periclinal divisions, vascular tissue gradually disappears in bHLH-deficient mutants; conversely, ectopic expression is sufficient for triggering periclinal divisions. We show that this dimer operates independently of tissue identity but is restricted to a small vascular domain by integrating overlapping transcription patterns of the interacting bHLH proteins. Our work reveals a common mechanism for tissue establishment and indeterminate vascular development and provides a conceptual framework for developmental control of local cell divisions.

Global biomass burning: a synthesis and review of Holocene paleofire records and their controls

We synthesize existing sedimentary charcoal records to reconstruct Holocene fire history at regional, continental and global scales. The reconstructions are compared with the two potential controls of burning at these broad scales – changes in climate and human activities – to assess their relative importance on trends in biomass burning. Here we consider several hypotheses that have been advanced to explain the Holocene record of fire, including climate, human activities and synergies between the two. Our results suggest that 1) episodes of high fire activity were relatively common in the early Holocene and were consistent with climate changes despite low global temperatures and low levels of biomass burning globally; 2) there is little evidence from the paleofire record to support the Early Anthropocene Hypothesis of human modification of the global carbon cycle; 3) there was a nearly-global increase in fire activity from 3 to 2 ka that is difficult to explain with either climate or humans, but the widespread and synchronous nature of the increase suggests at least a partial climate forcing; and 4) burning during the past century generally decreased but was spatially variable; it declined sharply in many areas, but there were also large increases (e.g., Australia and parts of Europe). Our analysis does not exclude an important role for human activities on global biomass burning during the Holocene, but instead provides evidence for a pervasive influence of climate across multiple spatial and temporal scales.

The pollen tube: a soft shell with a hard core

Plant cell expansion is controlled by a fine-tuned balance between intracellular turgor pressure, cell wall loosening and cell wall biosynthesis. To understand these processes, it is important to gain in-depth knowledge of cell wall mechanics. Pollen tubes are tip-growing cells that provide an ideal system to study mechanical properties at the single cell level. With the available approaches it was not easy to measure important mechanical parameters of pollen tubes, such as the elasticity of the cell wall. We used a cellular force microscope (CFM) to measure the apparent stiffness of lily pollen tubes. In combination with a mechanical model based on the finite element method (FEM), this allowed us to calculate turgor pressure and cell wall elasticity, which we found to be around 0.3 MPa and 20–90 MPa, respectively. Furthermore, and in contrast to previous reports, we showed that the difference in stiffness between the pollen tube tip and the shank can be explained solely by the geometry of the pollen tube. CFM, in combination with an FEM-based model, provides a powerful method to evaluate important mechanical parameters of single, growing cells. Our findings indicate that the cell wall of growing pollen tubes has mechanical properties similar to rubber. This suggests that a fully turgid pollen tube is a relatively stiff, yet flexible cell that can react very quickly to obstacles or attractants by adjusting the direction of growth on its way through the female transmitting tissue.

Soft and hard tissue histologic dimensions around dental implants in the canine restored with smaller-diameter abutments: a paradigm shift in peri-implant biology

PURPOSE To evaluate the biologic width dimensions around implants with nonmatching implant-abutment diameters. MATERIALS AND METHODS Five canines had their mandibular premolars and first molars removed bilaterally and replaced with 12 implants that had nonmatching implant-abutment diameters. On one side, six implants were placed in a submerged surgical approach, and the other side utilized a nonsubmerged approach. Two of the implants on each side were placed either 1 mm above, even with, or 1 mm below the alveolar crest. Two months later, gold crowns were attached, and the dogs were sacrificed 6 months postloading. Block sections were processed for histologic and histomorphometric analyses. RESULTS The bone level, connective tissue length, epithelial dimension, and biologic width were not significantly different when the implants were initially placed in a submerged or nonsubmerged surgical approach. The bone level was significantly different around implants placed 1 mm above the crest compared to implants placed even with or 1 mm below the alveolar crest. The connective tissue dimension was not different for any implant level placement. The epithelial dimension and biologic width were significantly greater for implants placed 1 mm below the alveolar crest compared to implants placed even with or 1 mm above the alveolar crest. For five of six implant placements, connective tissue covered the implant/abutment interface. CONCLUSIONS This study reveals a fundamental change in the biologic response to implants with nonmatching implant-abutment diameters. Unlike implants with matching implant-abutment diameters, the connective tissue extended coronally past the interface (microgap). This morphologic tissue alteration represents a significant change in the biologic reaction to implant-abutment interfaces and suggests that marginal inflammation is eliminated or greatly reduced in these implant designs.

Interaction between Meristem Tissue Layers Controls Phyllotaxis

Phyllotaxis and vein formation are among the most conspicuous patterning processes in plants. The expression and polarization of the auxin efflux carrier PIN1 is the earliest marker for both processes, with mathematical models indicating that PIN1 can respond to auxin gradients and/or auxin flux. Here, we use cell-layer-specific PIN1 knockouts and partial complementation of auxin transport mutants to examine the interaction between phyllotactic patterning, which occurs primarily in the L1 surface layer of the meristem, and midvein specification in the inner tissues. We show that PIN1 expression in the L1 is sufficient for correct organ positioning, as long as the L1-specific influx carriers are present. Thus, differentiation of inner tissues can proceed without PIN1 or any of the known polar transporters. On theoretical grounds, we suggest that canalization of auxin flux between an auxin source and an auxin sink may involve facilitated diffusion rather than polar transport.

A scrutiny of hard pion chiral perturbation theory

In the recently proposed framework of hard pion chiral perturbation theory, the leading chiral logarithms are predicted to factorize with respect to the energy dependence in the chiral limit. We have scrutinized this assumption in the case of vector and scalar pion form factors FV;S(s) by means of standard chiral perturbation theory and dispersion relations. We show that this factorization property is valid for the elastic contribution to the dispersion integrals for FV;S(s) but it is violated starting at three loops when the inelastic four-pion contributions arise.

Accelerometry and Energy Expenditure in Obese Adults and Normal Weight Controls

Objective: To evaluate a new triaxial accelerometer device for prediction of energy expenditure, measured as VO2/kg, in obese adults and normal-weight controls during activities of daily life. Subjects and methods: Thirty-seven obese adults (Body Mass Index (BMI) 37±5.4) and seventeen controls (BMI 23±1.8) performed eight activities for 5 to 8 minutes while wearing a triaxial accelerometer on the right thigh. Simultaneously, VO2 and VCO2 were measured using a portable metabolic system. The relationship between accelerometer counts (AC) and VO2/kg was analysed using spline regression and linear mixed-effects models. Results: For all activities, VO2/kg was significantly lower in obese participants than in normalweight controls. A linear relationship between AC and VO2/kg existed only within accelerometer values from 0 to 300 counts/min, with an increase of 3.7 (95%-confidence interval (CI) 3.4 - 4.1) and 3.9 ml/min (95%-CI 3.4 - 4.3) per increase of 100 counts/min in obese and normal-weight adults, respectively. Linear modelling of the whole range yields wide prediction intervals for VO2/kg of ± 6.3 and ±7.3 ml/min in both groups. Conclusion: In obese and normal-weight adults, the use of AC for predicting energy expenditure, defined as VO2/kg, from a broad range of physical activities, characterized by varying intensities and types of muscle work, is limited.