Sex of muscle stem cells does not influence potency for cardiac cell therapy

We have previously shown that populations of skeletal muscle-derived stem cells (MDSCs) exhibit sexbased differences for skeletal muscle and bone repair, with female cells demonstrating superior engrafting abilities to males in skeletal muscle while male cells differentiating more robustly toward the osteogenic and chondrogenic lineages. In this study, we tested the hypothesis that the therapeutic capacity of MDSCs transplanted into myocardium is influenced by sex of donor MDSCs or recipient. Male and female MDSCs isolated from the skeletal muscle of 3-week-old mice were transplanted into recipient male or female dystrophin-deficient (mdx) hearts or into the hearts of male SCID mice following acute myocardial infarction. In the mdx model, no difference was seen in engraftment or blood vessel formation based on donor cell or recipient sex. In the infarction model, MDSC-transplanted hearts showed higher postinfarction angiogenesis, less myocardial scar formation, and improved cardiac function compared to vehicle controls. However, sex of donor MDSCs had no significant effects on engraftment, angiogenesis, and cardiac function. VEGF expression, a potent angiogenic factor, was similar between male and female MDSCs. Our results suggest that donor MDSC or recipient sex has no significant effect on the efficiency of MDSC-triggered myocardial engraftment or regeneration following cardiac injury. The ability of the MDSCs to improve cardiac regeneration and repair through promotion of angiogenesis without differentiation into the cardiac lineage may have contributed to the lack of sex difference observed in these models. Copyright © 2009 Cognizant Comm. Corp

The role of cardiac troponin T quantity and function in cardiac development and dilated cardiomyopathy

Background: Hypertrophic (HCM) and dilated (DCM) cardiomyopathies results from sarcomeric protein mutations, including cardiac troponin T (cTnT, TNNT2). We determined whether TNNT2 mutations cause cardiomyopathies by altering cTnT function or quantity; whether the severity of DCM is related to the ratio of mutant to wildtype cTnT; whether Ca2+ desensitization occurs in DCM; and whether absence of cTnT impairs early embryonic cardiogenesis. Methods and Findings: We ablated Tnnt2 to produce heterozygous Tnnt2+/ mice, and crossbreeding produced homozygous null Tnnt2-/-embryos. We also generated transgenic mice overexpressing wildtype (TGWT) or DCM mutant (TGK210Δ) Tnnt2. Crossbreeding produced mice lacking one allele of Tnnt2, but carrying wildtype (Tnnt2+/-/TGWT) or mutant (Tnnt2+/-/TGK210Δ) transgenes. Tnnt2+/-mice relative to wildtype had significantly reduced transcript (0.82 ± 0.06 [SD] vs. 1.00 ± 0.12 arbitrary units; p = 0.025), but not protein (1.01 ± 0.20 vs. 1.00 ± 0.13 arbitrary units; p = 0.44). Tnnt2+/-mice had normal hearts (histology, mass, left ventricular end diastolic diameter [LVEDD], fractional shortening [FS]). Moreover, whereas Tnnt2+/-/ TGK210Δ mice had severe DCM, TGK210Δ mice had only mild DCM (FS 18 ± 4 vs. 29 ± 7%; p < 0.01). The difference in severity of DCM may be attributable to a greater ratio of mutant to wildtype Tnnt2 transcript in Tnnt2+/-/TGK210Δ relative to TGK210Δ mice (2.42±0.08, p = 0.03). Tnnt2+/-/TGK210Δ muscle showed Ca2+ desensitization (pCa50 = 5.34 ± 0.08 vs. 5.58 ± 0.03 at sarcomere length 1.9 μm. p<0.01), but no difference in maximum force generation. Day 9.5 Tnnt2-/-embryos had normally looped hearts, but thin ventricular walls, large pericardial effusions, noncontractile hearts, and severely disorganized sarcomeres. Conclusions: Absence of one Tnnt2 allele leads to a mild deficit in transcript but not protein, leading to a normal cardiac phenotype. DCM results from abnormal function of a mutant protein, which is associated with myocyte Ca2+ desensitization. The severity of DCM depends on the ratio of mutant to wildtype Tnnt2 transcript. cTnT is essential for sarcomere formation, but normal embryonic heart looping occurs without contractile activity. © 2008 Ahmad et al

Photoelectric Emission from Interstellar Dust: Grain Charging and Gas Heating

We model the photoelectric emission from and charging of interstellar dust and obtain photoelectric gas heating efficiencies as a function of grain size and the relevant ambient conditions. Using realistic grain size distributions, we evaluate the net gas heating rate for various interstellar environments, and find less heating for dense regions characterized by R_V=5.5 than for diffuse regions with R_V=3.1. We provide fitting functions which reproduce our numerical results for photoelectric heating and recombination cooling for a wide range of interstellar conditions. In a separate paper we will examine the implications of these results for the thermal structure of the interstellar medium. Finally, we investigate the potential importance of photoelectric heating in H II regions, including the warm ionized medium. We find that photoelectric heating could be comparable to or exceed heating due to photoionization of H for high ratios of the radiation intensity to the gas density. We also find that photoelectric heating by dust can account for the observed variation of temperature with distance from the galactic midplane in the warm ionized medium.Comment: 50 pages, including 18 figures; corrected title and abstract field

Developing cardiac and skeletal muscle share fast-skeletal myosin heavy chain and cardiac troponin-I expression

Skeletal muscle derived stem cells (MDSCs) transplanted into injured myocardium can differentiate into fast skeletal muscle specific myosin heavy chain (sk-fMHC) and cardiac specific troponin-I (cTn-I) positive cells sustaining recipient myocardial function. We have recently found that MDSCs differentiate into a cardiomyocyte phenotype within a three-dimensional gel bioreactor. It is generally accepted that terminally differentiated myocardium or skeletal muscle only express cTn-I or sk-fMHC, respectively. Studies have shown the presence of non-cardiac muscle proteins in the developing myocardium or cardiac proteins in pathological skeletal muscle. In the current study, we tested the hypothesis that normal developing myocardium and skeletal muscle transiently share both sk-fMHC and cTn-I proteins. Immunohistochemistry, western blot, and RT-PCR analyses were carried out in embryonic day 13 (ED13) and 20 (ED20), neonatal day 0 (ND0) and 4 (ND4), postnatal day 10 (PND10), and 8 week-old adult female Lewis rat ventricular myocardium and gastrocnemius muscle. Confocal laser microscopy revealed that sk-fMHC was expressed as a typical striated muscle pattern within ED13 ventricular myocardium, and the striated sk-fMHC expression was lost by ND4 and became negative in adult myocardium. cTn-I was not expressed as a typical striated muscle pattern throughout the myocardium until PND10. Western blot and RT-PCR analyses revealed that gene and protein expression patterns of cardiac and skeletal muscle transcription factors and sk-fMHC within ventricular myocardium and skeletal muscle were similar at ED20, and the expression patterns became cardiac or skeletal muscle specific during postnatal development. These findings provide new insight into cardiac muscle development and highlight previously unknown common developmental features of cardiac and skeletal muscle. © 2012 Clause et al

Collagen I weakly interacts with the β-sheets of β2-microglobulin and enhances conformational exchange to induce amyloid formation

Amyloidogenesis is significant in both protein function and pathology. Amyloid formation of folded, globular proteins is commonly initiated by partial or complete unfolding. However, how this unfolding event is triggered for proteins that are otherwise stable in their native environments is not well understood. The accumulation of the immunoglobulin protein β2-microglobulin (β2m) into amyloid plaques in the joints of long-term hemodialysis patients is the hallmark of Dialysis Related Amyloidosis (DRA). While β2m does not form amyloid unassisted near neutral pH in vitro, the localization of β2m deposits to joint spaces suggests a role for the local extracellular matrix (ECM) proteins, specifically collagens, in promoting amyloid formation. Indeed, collagen and other ECM components have been observed to facilitate β2m amyloid formation, but the large size and anisotropy of the complex, combined with the low affinity of these interactions, has limited atomic-level elucidation of the amyloid-promoting mechanism(s) by these molecules. Using solution NMR approaches that uniquely probe weak interactions in large molecular weight complexes, we are able to map the binding interfaces on β2m for collagen I and detect collagen I-induced μs–ms timescale dynamics in the β2m backbone. By combining solution NMR relaxation methods and 15N-dark state exchange saturation transfer experiments, we propose a model in which weak, multimodal collagen I–β2m interactions promote exchange with a minor population of amyloid-competent species to induce fibrillogenesis. The results portray the intimate role of the environment in switching an innocuous protein into an amyloid-competent state, rationalizing the localization of amyloid deposits in DRA

Estimation of Nitrogen Fixation by the Natural 15N-abundance Technique and Nitrogen Uptake by Pigeonpea Genotypes of Different Maturity Groups grown in an Inceptisol

The modulation, nitrogen fixation and nitrogen uptake of four pigeonpea genotypes belonging to extra short duration, short duration and long duration maturity groups grown on an Inceptisol were studied to examine why, despite the poor nodulation of pigeonpea in this soil, it still produces greater yields, than in Alfisols and Vertisols. The percentage nitrogen derived from the atmosphere (%Ndfa) was estimated by 15N natural abundance and N-difference methods using a long duration sorghum as the non-fixing reference crop. In general, nodulation of pigeonpea in the Inceptisol was much lower than that reported in Alfisols and Vertisols. The above-ground dry matter ringed from 3.1 to 17.1 t ha−1 while the N uptake ranged from 62.3 to 215 kg ha−1 The fallen plant parrs of pigeonpea genotypes ranged from 1.4 to 4.9 t ha−1 and their N contents ranged from 25 to 84 kg ha−1. The estimates of percentage Ndfa obtained by the two methods were different. Those obtained by the 15N natural abundance appeared more appropriate as the δ15N of sorghum harvested along with short duration pigeonpea and later when it was mature did not change significantly. The extra short duration pigeonpea genotype ICPL 84023 contained very little N from atmospheric N2, while the short duration pigeonpea cv. ICPL 151 had 17% Ndfa and the long duration genotypes. ICPL 366 and T7 had up to 36 % Ndfa. It can be concluded that one of the causes of high yields of pigeonpea on Inceptisols compared to Alfisols and Vertisols despite poor nodulation could be the high N supplying capacity of these Inceptisols. Strategies have been suggested as to how pigeonpea genotypes grown in Inceptisols could improve their nodulation and nitrogen fixation and thus better contribute to a sustainable agriculture

Modeling of synthesis and flow properties of propylene-diene copolymers

Copolymerization with nonconjugated dienes offers an attractive route for introducing long-chain branching in polypropylene. From a simplified set of rate equations for such copolymerization with a metallocene catalyst, we derive the probabilities of branch formation at different stages of the reaction in a semibatch reactor. Using these probabilities, we generate an ensemble of molecules via a Monte Carlo sampling. The knowledge of the branching topology and segment lengths allows us to compute the flow properties of the resins from computational rheology. We compare our model predictions with existing experimental data, namely the molar mass distribution and small amplitude oscillatory shear response, for a set of resins with varying diene content. The rheology data suggest that the entanglement time Ï.,e depends sensitively and in a well-defined fashion on the diene content

Field evaluation of nitrogen fixation and use of nitrogen fertilizer by sorghum/pigeonpea intercropping on an Alfisol in the Indian semi-arid tropics

A field experiment was conducted to obtain the N balance sheet for sole crops and intercrops of sorghum [Sorghum bicolor (L.) Moench] and pigeonpeas [Cajanus cajan (L.) Millsp.]. Intercropping gave a significant advantage over sole cropping in terms of dry matter production and grain yield, as calculated on the basis of the land equivalent ratio and area-time equivalent ratio. The N fertilizer use efficiency and atmospheric N2 fixation by pigeonpea were estimated using 15N-labeling and natural abundance methods. The N fertilizer use efficiency of sorghum was unaltered by the cropping system, while that of the pigeonpea was greatly reduced by intercropping. Although intercropping increased the fractional contribution of fixed N to the pigeonpeas, no significant difference was observed between the cropping systems in total symbiotically fixed N. There was no evidence of a significant transfer of N from the pigeonpea to the sorghum. This study showed that use of soil N and fertilizer N by pigeonpeas was almost the same as that by sorghum in sole cropping, indicating the potential competence of pigeonpeas to exploit soil N. However, when N was exhausted by a companion crop in intercropping, the pigeonpea crop increased its dependency on atmospheric N2 fixation. We conclude that knowledge of how N from different sources is shared by companion crops is a prerequisite to establishing strategies to increase N use, and consequently land productivity, in intercropping systems