Direct isolation of satellite cells for skeletal muscle regeneration

Muscle satellite cells contribute to muscle regeneration. We have used a Pax3(GFP/+) mouse line to directly isolate (Pax3)(green fluorescent protein)-expressing muscle satellite cells, by flow cytometry from adult skeletal muscles, as a homogeneous population of small, nongranular, Pax7+, CD34+, CD45-, Sca1- cells. The flow cytometry parameters thus established enabled us to isolate satellite cells from wild-type muscles. Such cells, grafted into muscles of mdx nu/nu mice, contributed both to fiber repair and to the muscle satellite cell compartment. Expansion of these cells in culture before engraftment reduced their regenerative capacity.     

G protein-coupled receptors in Anopheles gambiae

We used bioinformatic approaches to identify a total of 276 G protein-coupled receptors (GPCRs) from the Anopheles gambiae genome. These include GPCRs that are likely to play roles in pathways affecting almost every aspect of the mosquito's life cycle. Seventy-nine candidate odorant receptors were characterized for tissue expression and, along with 76 putative gustatory receptors, for their molecular evolution relative to Drosophila melanogaster. Examples of lineage-specific gene expansions were observed as well as a single instance of unusually high sequence conservation.     

Peroxisomal defects in neonatal-onset and X-linked adrenoleukodystrophies

Accumulation of very long chain fatty acids in X-linked and neonatal forms of adrenoleukodystrophy (ALD) appears to be a consequence of deficient peroxisomal oxidation of very long chain fatty acids. Peroxisomes were readily identified in liver biopsies taken from a patient having the X-linked disorder. However, in liver biopsies from a patient having neonatal-onset ALD, hepatocellular peroxisomes were greatly reduced in size and number, and sedimentable catalase was markedly diminished. The presence of increased concentrations of serum pipecolic acid and the bile acid intermediate, trihydroxycoprostanic acid, in the neonatal ALD patient are associated with a generalized diminution of peroxisomal activities that was not observed in the patient with X-linked ALD.     

Control of embryonic motoneuron survival in vivo by ciliary neurotrophic factor

During development of the nervous system, neurons in many regions are overproduced by proliferation, after which the excess cells are eliminated by cell death. The survival of only a proportion of neurons during normal development is thought to be regulated by the limited availability of neurotrophic agents. One such putative trophic agent is ciliary neurotrophic factor (CNTF), a polypeptide that promotes the survival of ciliary, sensory, and sympathetic neurons in vitro. In contrast to the results of in vitro studies, however, the daily treatment of chick embryos in vivo with purified human recombinant CNTF failed to rescue any of these cell populations from cell death, whereas CNTF did promote the in vivo survival of spinal motoneurons. Thus, CNTF may not act as a neurotrophic agent in vivo for those embryonic neurons (especially ciliary neurons) on which it acts in vitro. Rather, CNTF may be required for in vivo survival of motoneurons.     

Conductance-controlled point functionalization of single-walled carbon nanotubes

We used covalent attachments to single-walled carbon nanotubes (SWNTs) to fabricate single-molecule electronic devices. The technique does not rely on submicrometer lithography or precision mechanical manipulation, but instead uses circuit conductance to monitor and control covalent attachment to an electrically connected SWNT. Discrete changes in the circuit conductance revealed chemical processes happening in real time and allowed the SWNT sidewalls to be deterministically broken, reformed, and conjugated to target species. By controlling the chemistry through electronically controlled electrochemical potentials, we were able to achieve single chemical attachments. We routinely functionalized pristine, defect-free SWNTs at one, two, or more sites and demonstrated three-terminal devices in which a single attachment controls the electronic response.     

Lack of correlation between the levels of soluble cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) and the CT-60 genotypes

Background

Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) plays a critical role in downregulation of antigen-activated immune response and polymorphisms at the CTLA-4 gene have been shown to be associated with several autoimmune diseases including type-1 diabetes (T1D). The etiological mutation was mapped to the CT60-A/G single nucleotide polymorphism (SNP) that is believed to control the processing and production of soluble CTLA-4 (sCTLA-4).

Methods

We therefore determined sCTLA-4 protein levels in the sera from 82 T1D patients and 19 autoantibody positive (AbP) subjects and 117 autoantibody negative (AbN) controls using ELISA. The CT-60 SNP was genotyped for these samples by using PCR and restriction enzyme digestion of a 268 bp DNA segment containing the SNP. Genotyping of CT-60 SNP was confirmed by dye terminating sequencing reaction.

Results

Higher levels of sCTLA-4 were observed in T1D (2.24 ng/ml) and AbP (mean = 2.17 ng/ml) subjects compared to AbN controls (mean = 1.69 ng/ml) with the differences between these subjects becoming significant with age (p = 0.02). However, we found no correlation between sCTLA-4 levels and the CTLA-4 CT-60 SNP genotypes.

Conclusion

Consistent with the higher serum sCTLA-4 levels observed in other autoimmune diseases, our results suggest that sCTLA-4 may be a risk factor for T1D. However, our results do not support the conclusion that the CT-60 SNP controls the expression of sCTLA-4.

     

Antibody recognition to secreted proteins of Mycobacterium avium subsp. paratuberculosis in sera from infected ruminants

Two liquid culture media to obtain secreted proteins of Mycobacterium avium subsp. paratuberculosis at different incubation periods were evaluated. Middlebrook 7H9-OADC (7H9) and Watson-Reid (WR) broths were inoculated with a field strain of M. paratuberculosis and growth curves determined using nonlinear regression analysis. Most culture filtrate (CF) proteins were of low molecular weight and reacted strongly against sera from cultured-positive cases of paratuberculosis. CF proteins obtained in WR yielded a higher number of bands and were detected earlier than those obtained from 7H9. A high degree of variability in CF protein immunoreactivity was seen among infected animals. Sera from cattle with clinical paratuberculosis or heavy fecal shedders of M. paratuberculosis reacted more intensively and to more CF proteins than did sera from other infected cattle. Immunoblots showed differences in antibody binding to CF proteins when sera were absorbed with M. avium but not with others environmental mycobacteria. Immunoblots with sera from infected goats and a sheep showed reactivity with proteins of 32, 33 and 46 kDa both before and after the sera were absorbed with M. phlei. Antibodies found in serum of infected deer reacted with CF proteins in a similar way as did for cattle. These results suggest that a pool of CF proteins of M. paratuberculosis could be good candidates as antigens for serodiagnosis of paratuberculosis.     

Effects of dietary Camelina sativa products on digestible nutrient compositions for rainbow trout (Oncorhynchus mykiss)

Two trials were conducted to determine the effect of dietary inclusion of camelina (Camelina sativa) products on their digestible nutrient compositions in rainbow trout. In the first experiment, the following camelina products: full‐fat seed, oil, high‐oil residue meal (HORM), pre‐press solvent‐extracted meal (SECM) and toasted SECM were evaluated. In the second trial, the utilization of SECM, SECM soaked in water (WS), SECM treated with a multicarbohydrase, Superzyme?‐OM, SECM treated with Bio‐Phytase and SECM treated with Superzyme?‐OM and Bio‐Phytase (MIX) was determined. The experimental diets consisted of a basal diet and test ingredient with a ratio of 70:30, except for camelina oil where the ratio was 80:20. In Experiment 1, SECM had the highest level of digestible crude protein (352 g kg^?1), whereas the oil was the best source of digestible energy (8,063 kcal kg^?1) and digestible crude fat (908 g kg^?1). In Experiment 2, there were no significant differences in nutrient digestibility of SECM and HORM; however, WS and MIX had significantly more digestible dry matter and digestible crude fat than SECM and HORM. Camelina by‐products have the potential to replace fishery by‐products in rainbow trout feeds.     

不同耕作方式对宁南旱地土壤团聚体的影响

在宁南旱区对试验耕地定位实行连年传统翻耕(CT)、隔年免耕—深松(NT→ST)及深松—免耕(ST→NT)三种不同处理。用干筛法和湿筛法对连续3年处理后土壤团聚体的变化进行测定,分析结果表明,干筛法中NT→ST处理在0~40 cm土层中0.25 mm的机械稳定性团聚体比处理前和CT处理分别高33.11%、29.51%;ST→NT处理在0~40 cm土层中0.25 mm机械稳定性团聚体含量比处理前和CT处理分别高40.51%、36.91%。湿筛法中NT→ST处理在0~30 cm土层中0.25 mm的水稳性团聚体含量比处理前高7.33%,NT→ST处理在0~10 cm土层0.25 mm的水稳性团聚体比CT处理高1.47%,而在10~40 cm土层NT→ST处理和CT0.25 mm的水稳性团聚体含量差异不显著;ST→NT处理在0~40 cm土层中0.25 mm水稳性团聚体含量比处理前和CT处理分别高10.75%、10.12%。分析结果同时说明,采用深松—免耕(ST→NT)轮耕方式更有利于土壤团聚体含量和稳定性的增加。