Reduced transforming growth factor-beta signaling in cartilage of old mice: role in impaired repair capacity

Osteoarthritis (OA) is a common joint disease, mainly effecting the elderly population. The cause of OA seems to be an imbalance in catabolic and anabolic factors that develops with age. IL-1 is a catabolic factor known to induce cartilage damage, and transforming growth factor (TGF)-beta is an anabolic factor that can counteract many IL-1-induced effects. In old mice, we observed reduced responsiveness to TGF-beta-induced IL-1 counteraction. We investigated whether expression of TGF-beta and its signaling molecules altered with age. To mimic the TGF-beta deprived conditions in aged mice, we assessed the functional consequence of TGF-beta blocking. We isolated knee joints of mice aged 5 months or 2 years, half of which were exposed to IL-1 by intra-articular injection 24 h prior to knee joint isolation. Immunohistochemistry was performed, staining for TGF-beta1, -2 or -3, TGF-betaRI or -RII, Smad2, -3, -4, -6 and -7 and Smad-2P. The percentage of cells staining positive was determined in tibial cartilage. To mimic the lack of TGF-beta signaling in old mice, young mice were injected with IL-1 and after 2 days Ad-LAP (TGF-beta inhibitor) or a control virus were injected. Proteoglycan (PG) synthesis (³⁵S-sulfate incorporation) and PG content of the cartilage were determined. Our experiments revealed that TGF-beta2 and -3 expression decreased with age, as did the TGF-beta receptors. Although the number of cells positive for the Smad proteins was not altered, the number of cells expressing Smad2P strongly dropped in old mice. IL-1 did not alter the expression patterns. We mimicked the lack of TGF-beta signaling in old mice by TGF-beta inhibition with LAP. This resulted in a reduced level of PG synthesis and aggravation of PG depletion. The limited response of old mice to TGF-beta induced-IL-1 counteraction is not due to a diminished level of intracellular signaling molecules or an upregulation of intracellular inhibitors, but is likely due to an intrinsic absence of sufficient TGF-beta receptor expression. Blocking TGF-beta distorted the natural repair response after IL-1 injection. In conclusion, TGF-beta appears to play an important role in repair of cartilage and a lack of TGF-beta responsiveness in old mice might be at the root of OA development.     

The Physio1ogical Activity of 2 : 6-Substituted Phenoxyacetic Acids

Anumber of 2 : 6-substituted phenoxyacetic acids in which theside chain had been substituted by ailcyl groups to form -propionic,-n-butyric and -n-valeric acids were investigated to assesstheir ability to act as growth-regulators. Besides employingstandard tedmiques of bioassay, further experiments were undertaken to determine the effects of these compounds on the vegetativegrowth and development of intact plants of Helianthus annuus. It has been established that all the compounds tested inducedcurvature in the Went pea curvature test and that without exceptionthe activity of the parent phenoxyacetic acid (2:6-dichloro-,2:4:6-trichloro-,2:6-dimethyl-, and 2:4-dichloro-6-methyl-)was increased by side chain substitution. In the Avena straight growth assay, the a 2:6-dichloro- and2:6-dimethyl- phenoxypropionic and butyric acids brought aboutstatistically significant increases in length of the coleoptilesections, when measurements were made at the end of 24 hours.Their activity was of the same order as that exhibited by a2:4-dichlorophenoxyacetic acid. Investigations were carried out on the uptake of water by sectionsof pea internode and the extension growth of Avena coleoptileaectiona when both the concentration and the length of treatmentwere varied. For some compounds it was found that over a certainrange of concentration water uptake and extension growth wereaccelerated in the initial 6–8 hours. Subsequent to this,inhibition occurred so that no significant increases above controlvalues were apparent at the end of 24 hours, and in some casesan actual loss of water or shrinkage in length took place. Itwas thus possible to demonstrate that 2:6-dimethylphenoxyaceticacid acts as a growth regulator in pea extension growth andthat 2:6-dichiorophenoxyacetic acid is active in the Avena test. The changes in the leaf area of individual pairs of leaves,together with the lengths of the internodes and shoot weightwere followed after application of measured amounts of eachcompound to the first pair of leaves of H. annuus. A numberof the 2:6-compounds were capable of modifying growth in wayssimilar to 2:4-dichlorophenoxyacetlc acid. However, much higherconcentration had to be applied in order to produce effectscomparable to those caused by the 2:4-dichloro- compound. Measurementsof the percentage of applied compound which penetrated intothe leaf showed that there were no marked differences in thisrespect between the different phenoxy acids. On the other hand,experiments in which the treated leaves were left on the plantfor varying periods of time led to the conclusion that the a: 6-substituted compounds are less readily translocated than2:4-dichlorophenoxyacetic acid.     

Effect of weather on the reproductive rate of Rock Ptarmigan Lagopus muta in the eastern Pyrenees

Understanding the effects of climate on avian life history traits is essential if we wish to predict the demographic consequences of expected climatic changes. We investigated the influence of weather conditions on the reproductive success of Rock Ptarmigan Lagopus muta in the eastern French Pyrenees, one of the southernmost areas inhabited by the species. Reproductive success was estimated in early August between 1997 and 2006 by counting adults and well-grown chicks with pointing dogs. The number of young per adult varied from 0.08 to 0.72. Using Poisson regression and Akaike's information criterion, we selected the best model explaining the effect of weather (date of snowmelt and, for both laying/incubation and post-hatching periods, mean minimum and maximum temperatures, monthly rainfall and number of days with rain) on the proportion of young in August. Reproductive success was positively associated with early appearance of snow-free ground, and date of snowmelt alone was the model that best explained annual variation in reproductive success. Other models, which included a negative effect of rainfall, particularly after hatching, also had some support. Hence, both pre-laying and post-hatching weather conditions influenced reproductive success of Rock Ptarmigan in the eastern French Pyrenees. On a continental scale, reproductive success of alpine populations of Rock Ptarmigan is consistently lower than that of northern populations. This difference in productivity may be partly correlated with climatic conditions observed along an arctic–alpine gradient, the amount and variation of rainfall being greater in southern alpine areas than elsewhere in the species' range.     

Phylogenetic analysis of Chloraeinae (Orchidaceae) based on plastid and nuclear DNA sequences

The phylogenetic relationships of subtribe Chloraeinae, a group of terrestrial orchids endemic to southern South America, have not been satisfactorily investigated. A previous molecular phylogenetic analysis based on plastid DNA supported the monophyly of Chloraeinae and Gavilea, but showed that Chloraea is non‐monophyletic and that the sole species of Bipinnula analysed is sister to Geoblasta. However, that analysis included only 18 of the 73 species belonging to this subtribe. Here, the phylogenetic relationships of Chloraeinae were assessed by analysing aproximately 7500 bp of nucleotide sequences from nuclear ribosomal internal transcribed spacer (ITS) and plastid DNA (rbcL, matK, trnL‐trnF, rpoB‐trnC) for 42 species representing all four currently accepted genera of Chloraeinae and appropriate outgroups. Nuclear and plastid data were analysed separately and in combination using two different methods, namely parsimony and Bayesian inference. Our analyses support the monophyly of Chloraeinae and their inclusion in an expanded concept of Cranichideae, but none of the genera of Chloraeinae that includes more than one species is monophyletic. Gavilea and Bipinnula are paraphyletic, with Chloraea chica nested in Gavilea and Geoblasta penicillata in Bipinnula. As currently delimited, Chloraea is polyphyletic. The taxonomic changes proposed recently are for the most part not justifiable on phylogenetic grounds, except for recognition of the monotypic genus Correorchis. The lack of resolution for the relationships among species of ‘core’Chloraea suggests a relatively recent diversification of this group. The current generic classification is in need or revision, but additional study is advisable before carrying out further taxonomic changes. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 258–277.     

Use of chromomycin A3 staining in bovine sperm cells for detection of protamine deficiency

Sperm chromatin integrity is essential for accurate transmission of male genetic information, and normal sperm chromatin structure is important for fertilization. Protamine is a nuclear protein that plays a key role in sperm DNA integrity, because it is responsible for sperm DNA stability and packing until the paternal genome is delivered into the oocyte during fertilization. Our aim was to investigate protamine deficiency in sperm cells of Bos indicus bulls (Nelore) using chromomycin A₃ (CMA₃) staining. Frozen semen from 14 bulls were thawed, then fixed in Carnoy's solution. Smears were prepared and analyzed by microscopy. As a positive control of CMA₃ staining, sperm from one bull was subjected to deprotamination of nuclei. The percentage of CMA₃-positive bovine sperm did not vary among batches. Only two bulls showed a higher percentage of CMA₃-positive sperm cells compared to the others. CMA₃ is a simple and useful tool for detecting sperm protamine deficiency in bulls.     

The genome of Rhizobium leguminosarum has recognizable core and accessory components

Background

Rhizobium leguminosarum is an α-proteobacterial N₂-fixing symbiont of legumes that has been the subject of more than a thousand publications. Genes for the symbiotic interaction with plants are well studied, but the adaptations that allow survival and growth in the soil environment are poorly understood. We have sequenced the genome of R. leguminosarum biovar viciae strain 3841.

Results

The 7.75 Mb genome comprises a circular chromosome and six circular plasmids, with 61% G+C overall. All three rRNA operons and 52 tRNA genes are on the chromosome; essential protein-encoding genes are largely chromosomal, but most functional classes occur on plasmids as well. Of the 7,263 protein-encoding genes, 2,056 had orthologs in each of three related genomes (Agrobacterium tumefaciens, Sinorhizobium meliloti, and Mesorhizobium loti), and these genes were over-represented in the chromosome and had above average G+C. Most supported the rRNA-based phylogeny, confirming A. tumefaciens to be the closest among these relatives, but 347 genes were incompatible with this phylogeny; these were scattered throughout the genome but were over-represented on the plasmids. An unexpectedly large number of genes were shared by all three rhizobia but were missing from A. tumefaciens.

Conclusion

Overall, the genome can be considered to have two main components: a 'core', which is higher in G+C, is mostly chromosomal, is shared with related organisms, and has a consistent phylogeny; and an 'accessory' component, which is sporadic in distribution, lower in G+C, and located on the plasmids and chromosomal islands. The accessory genome has a different nucleotide composition from the core despite a long history of coexistence.     

Carbon and nitrogen content based estimation of the fat content of animal carcasses in various species

The objective of this study was to explore whether the C and N content can be used to estimate the fat content of animal carcasses. Considering the mean C and N contents of body fat and body protein, the fat content (EE) [%] can be predicted from C and N values [%] according to the generally valid equation EE=1.3038·C – 4.237·N. The application of this equation to estimate the total fat content of all animal carcasses results in significant differences in fat content between predicted and measured values. Therefore, we derived specific equations for rats, pigs, cattle, sheep, broilers and mice to predict the fat content by dual linear regression analysis (y=EE [% DM], x₁=C [% DM], x₂=N [% DM]) based on measured fat, C, and N contents of animal body samples. The specific equations for different animals showed residual standard deviations of 1.55, 1.63, 1.12, 1.35, 1.85 and 0.92% fat for rats, pigs, cattle, sheep, broilers and mice, respectively.