Tyrosine phosphorylation of myosin heavy chain during skeletal muscle differentiation: an integrated bioinformatics approach

Previously it has been shown that insulin-mediated tyrosine phosphorylation of myosin heavy chain is concomitant with enhanced association of C-terminal SRC kinase during skeletal muscle differentiation. We sought to identify putative site(s) for this phosphorylation event. A combined bioinformatics approach of motif prediction and evolutionary and structural analyses identified tyrosines163 and 1856 of the skeletal muscle heavy chain as the leading candidate for the sites of insulin-mediated tyrosine phosphorylation. Our work is suggestive that tyrosine phosphorylation of myosin heavy chain, whether in skeletal muscle or in platelets, is a significant event that may initiate cytoskeletal reorganization of muscle cells and platelets. Our studies provide a good starting point for further functional analysis of MHC phosphor-signalling events within different cells.     

Autocrine regulation of internal anal sphincter tone by renin-angiotensin system: comparison with phasic smooth muscle

The myogenic control mechanisms that govern the basal tone in the internal anal sphincter (IAS) are not known. The present studies determined the autocrine regulation of ANG II in the IAS. The studies were performed in the freshly isolated smooth muscle cells (SMC) of the IAS. We determined the presence of ANG II precursor angiotensinogen (Angen), and the enzymes that convert it into ANG II, using functional, molecular biology, and immunocytochemical studies in rats. ANG II levels in the SMC were determined using ELISA. The IAS SMC generate ANG II at a rate severalfold higher than those from the adjoining smooth muscle of rectum (RSM). RT-PCR data show that IAS exclusively expresses significant higher levels of renin, Angen, and angiotensin-converting enzyme (ACE). These data were confirmed using Western blot analyses and immunocytochemistry. In the IAS SMC, H-77 (10 microM; renin inhibitor) and captopril (1 microM; ACE inhibitor) decreased the basal as well as Angen-increased levels of ANG II. The following functional data corroborate the role of renin-angiotensin system (RAS) in the IAS tone. Angen produced concentration-dependent shortening of the IAS SMC that was inhibited by H-77 and captopril. In addition, H-77 or captopril caused a concentration-dependent fall in the IAS tone vs. nontonic tissues. Basal tone in IAS is partially under the autocrine control of cellular RAS evident by the expression of mRNA coding Angen, renin, and ACE and translation to the respective proteins in the SMC.     

Microsurgical reconstruction of the esophagus using supercharged pedicled jejunum flaps: special indications and pitfalls

This report outlines a microsurgical technique for total esophageal reconstruction in situations in which conventional methods using stomach or colon are not available. Eleven patients with corrosive injury and one patient following tumor resection underwent total esophageal reconstruction in a two-stage procedure. In the first stage, skin flaps or free jejunal transfers were used for the cervical reconstruction. In the second stage, supercharged pedicled jejunum flaps placed subcutaneously were used for thoracic esophageal replacement. The study included one male and 10 female patients, with a mean age of 38.4 years. The mean follow-up period was 78.9 months. All patients had one or more complications that required revisional surgery. Pedicled myocutaneous flaps were used to close fistulas or chronic wounds in four patients. The cervical skin tube in two patients and the jejunum in another two patients required shortening because of redundancy. Four patients had dysphagia caused by neck contractures, which were released. Two patients developed pharyngoesophageal strictures that required further free skin flaps for release. Two patients had reflux because of blind pouches arising from the original esophagus and required thoracotomy for removal. At long-term follow-up, all patients are fully rehabilitated and have resumed an oral diet with significant weight gain. Compared with lifelong jejunostomy feeding and its associated psychosocial disadvantages, the authors' experience demonstrates that the application of microsurgical techniques to fully reconstruct the esophagus is of considerable benefit to this difficult patient group.     

Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells

Human bone marrow stromal cells (hMSCs) were stably transduced by a retroviral vector containing the gene for the catalytic subunit of human telomerase (hTERT). Transduced cells (hMSC-TERTs) had telomerase activity, and the mean telomere length was increased as compared with that of control cells. The transduced cells have now undergone more than 260 population doublings (PD) and continue to proliferate, whereas control cells underwent senescence-associated proliferation arrest after 26 PD. The cells maintained production of osteoblastic markers and differentiation potential during continuous subculturing, did not form tumors, and had a normal karyotype. When implanted subcutaneously in immunodeficient mice, the transduced cells formed more bone than did normal cells. These results suggest that ectopic expression of telomerase in hMSCs prevents senescence-associated impairment of osteoblast functions.     

Two groups control light-induced schiff base deprotonation and the proton affinity of asp(85) in the Arg(82)His mutant of bacteriorhodopsin

Arg(82) is one of the four buried charged residues in the retinal binding pocket of bacteriorhodopsin (bR). Previous studies show that Arg(82) controls the pK(a)s of Asp(85) and the proton release group and is essential for fast light-induced proton release. To further investigate the role of Arg(82) in light-induced proton pumping, we replaced Arg(82) with histidine and studied the resulting pigment and its photochemical properties. The main pK(a) of the purple-to-blue transition (pK(a) of Asp(85)) is unusually low in R82H: 1.0 versus 2.6 in wild type (WT). At pH 3, the pigment is purple and shows light and dark adaptation, but almost no light-induced Schiff base deprotonation (formation of the M intermediate) is observed. As the pH is increased from 3 to 7 the M yield increases with pK(a) 4.5 to a value approximately 40% of that in the WT. A transition with a similar pK(a) is observed in the pH dependence of the rate constant of dark adaptation, k(da). These data can be explained, assuming that some group deprotonates with pK(a) 4.5, causing an increase in the pK(a) of Asp(85) and thus affecting k(da) and the yield of M. As the pH is increased from 7 to 10.5 there is a further 2.5-fold increase in the yield of M and a decrease in its rise time from 200 &mgr;s to 75 &mgr;s with pK(a) 9. 4. The chromophore absorption band undergoes a 4-nm red shift with a similar pK(a). We assume that at high pH, the proton release group deprotonates in the unphotolyzed pigment, causing a transformation of the pigment into a red-shifted "alkaline" form which has a faster rate of light-induced Schiff base deprotonation. The pH dependence of proton release shows that coupling between Asp(85) and the proton release group is weakened in R82H. The pK(a) of the proton release group in M is 7.2 (versus 5.8 in the WT). At pH < 7, most of the proton release occurs during O --> bR transition with tau approximately 45 ms. This transition is slowed in R82H, indicating that Arg(82) is important for the proton transfer from Asp(85) to the proton release group. A model describing the interaction of Asp(85) with two ionizable residues is proposed to describe the pH dependence of light-induced Schiff base deprotonation and proton release.     

Some unusual nucleic acid bases are products of hydroxyl radical oxidation of DNA and RNA

There are over 100 modified bases and their derivatives found in RNA and DNA. For some of them, data concerning their properties, synthesis and roles in cellular metabolism are available, but for others the knowledge of their functions and biosynthetic pathways is rather limited. We have analysed the chemical structure of modified nucleosides of DNA and RNA considering mainly their putative synthetic routes. On this basis we suggest, that in addition to enzymatic biosynthetic pathways well established for some odd bases, many rare nucleosides can be recognised as products of random chemical reactions. We identify them as primary or secondary products of the reaction of nucleic acids with hydroxyl radicals, the most active oxidising agent in the cell.     

Effects of soil erosion on crop productivity

Soil erosion and the effects of soil erosion on crop productivity have become emotional issues and have attracted the attention of agriculturists, environmentalists, and the public in general. In spite of heavy investments in research and development, the global rates of accelerated erosion are now presumbly higher than ever before. However, the data from available records obtained by diverse methods are uncomparable, unreliable, confusing, and often vary by several orders of magnitude. Reports of erosion‐caused alterations in crop productivity and soil properties are also contradictory and subjective. In addition to the lack of standardized methodology in evaluating soil erosion and its effects on crops, controversial interpretations are attributed to differences in soil profile characteristics, nutrient status, crops grown, and prevailing climatic conditions. Although erosion is generally associated wtih yield reductions, there are examples of where soil erosion has had no effect or has had a positive effect on crop production. Accelerated erosion affects productivity both directly and indirectly. Directly, the erosion‐induced reduction in crop yields is attributed to loss of rooting depth, degradation of soil structure, decrease in plant‐available water reserves, reduction in organic matter, and nutrient imbalance. Depending on soil properties and the degree of degradation, adverse effects of erosion on crop yields can be mostly compensated for by additional inputs of macronu‐trients (N, P, K) and macronutrients plus organic matter, by supplemental applications of some micronu‐trients, and by irrigation. For some soils, e.g., tropical soils, crop yields from severely eroded soils are significantly lower than those from uneroded lands and are often uneconomic in spite of additional inputs. Specific examples of yield alterations are given in relation to the loss of plant nutrients, soil water reserves, and alterations in soil properties. Criteria for soil‐loss tolerance are discussed, and productivity restoration of eroded soils is reviewed in relation to soil organic matter content and nutrient requirments. Research and development priorities are presented.     

Methane and nitrous oxide emissions under no‐till farming in China: a meta‐analysis

No‐till (NT) practices are among promising options toward adaptation and mitigation of climate change. However, the mitigation effectiveness of NT depends not only on its carbon sequestration potential but also on soil‐derived CH₄ and N₂O emissions. A meta‐analysis was conducted, using a dataset involving 136 comparisons from 39 studies in China, to identify site‐specific factors which influence CH₄ emission, CH₄ uptake, and N₂O emission under NT. Comparative treatments involved NT without residue retention (NT0), NT with residue retention (NTR), compared to plow tillage (PT) with residue removed (PT0). Overall, NT0 significantly decreased CH₄ emission by ~30% (P < 0.05) compared to PT0 with an average emission 218.8 kg ha⁻¹ for rice paddies. However, the increase in N₂O emission could partly offset the benefits of the decrease in CH₄ emission under NT compared to PT0. NTR significantly enhanced N₂O emission by 82.1%, 25.5%, and 20.8% (P < 0.05) compared to PT0 for rice paddies, acid soils, and the first 5 years of the experiments, respectively. The results from categorical meta‐analysis indicated that the higher N₂O emission could be mitigated by adopting NT within alkaline soils, for long‐term duration, and with less N fertilization input when compared to PT0. In addition, the natural log (lnR) of response ratio of CH₄ and N₂O emissions under NT correlated positively (enhancing emission) with climate factors (temperature and precipitation) and negatively (reducing emission) with experimental duration, suggesting that avoiding excess soil wetness and using NT for a long term could enhance the benefits of NT. Therefore, a thorough understanding of the conditions favoring greenhouse gas(es) reductions is essential to achieving climate change mitigation and advancing food security in China.     

Age-dependent behavioral changes and physiological changes in identified neurons in Aplysia californica

The gill withdrawal reflex (GWR) to direct gill stimulation was studied in sexually mature Aplysia and in those older by at least two months. The GWR threshold in old Aplysia was five- to sevenfold higher than that in mature animals. In the habituation paradigm, the GWR amplitude decremented rapidly to zero in old animals whereas in mature animals it persisted for at least ten trials. The GWR could not be dishabituated in old animals. The GWR is an age-dependent behavior in that parieto-visceral ganglion suppression of the GWR appears to increase with age. Also the electrophysiological properties of two neurons in the parieto-visceral ganglion were compared in the two age groups: L₇ a neuron which dishabituates the GWR in mature and not in old animals; and R₂ which manifests cytological changes with age. In old animals L₇′s input resistance was lower, the time constant was increased, and the size of the psp evoked by gill stimulation was smaller than those of mature L₇s. Similar membrane changes with age were measured in R₂. Soma size of L₇ was approximately the same in the two age groups as was that of R₂. The physiological parameters of neurons of known function continue to change during postmetamorphic life of Aplysia.