Trabecular bone ontogeny in the human proximal femur

Ontogenetic changes in the human femur associated with the acquisition of bipedal locomotion, especially the development of the bicondylar angle, have been well documented. The purpose of this study is to quantify changes in the three-dimensional structure of trabecular bone in the human proximal femur in relation to changing functional and external loading patterns with age. High-resolution X-ray computed tomography scan data were collected for 15 juvenile femoral specimens ranging in age from prenatal to approximately nine years of age. Serial slices were collected for the entire proximal femur of each individual with voxel resolutions ranging from 0.017 to 0.046 mm depending on the size of the specimen. Spherical volumes of interest were defined within the proximal femur, and the bone volume fraction, trabecular thickness, trabecular number, and fabric anisotropy were calculated in three dimensions. Bone volume fraction, trabecular number, and degree of anisotropy decrease between the age of 6 months and 12 months, with the lowest values for these parameters occurring in individuals near 12 months of age. By age 2-3 years, the bone volume, thickness, and degree of anisotropy increase slightly, and regions in the femoral neck become more anisotropic corresponding to the thickening of the inferior cortical bone of the neck. These results suggest that trabecular structure in the proximal femur reflects the shift in external loading patterns associated with the initiation of unassisted walking in infants.     

Genetic linkage and horizontal gene transfer, the roots of the antibiotic multi-resistance problem

Bacteria carrying resistance genes for many antibiotics are moving beyond the clinic into the community, infecting otherwise healthy people with untreatable and frequently fatal infections. This state of affairs makes it increasingly important that we understand the sources of this problem in terms of bacterial biology and ecology and also that we find some new targets for drugs that will help control this growing epidemic. This brief and eclectic review takes the perspective that we have too long thought about the problem in terms of treatment with or resistance to a single antibiotic at a time, assuming that dissemination of the resistance gene was affected by simple vertical inheritance. In reality antibiotic resistance genes are readily transferred horizontally, even to and from distantly related bacteria. The common agents of bacterial gene transfer are described and also one of the processes whereby nonantibiotic chemicals, specifically toxic metals, in the environment can select for and enrich bacteria with antibiotic multiresistance. Lastly, some speculation is offered on broadening our perspective on this problem to include drugs directed at compromising the ability of the mobile elements themselves to replicate, transfer, and recombine, that is, the three "infrastructure" processes central to the movement of genes among bacteria.     

FRET multiphoton spectral imaging microscopy of 7-ketocholesterol and Nile Red in U937 monocytic cells loaded with 7-ketocholesterol

OBJECTIVE: To show the effect of 7-ketocholesterol (7KC) on cellular lipid content by means of flow cytometry and the interaction of 7KC with Nile Red (NR) via ultraviolet fluorescence resonance energy transfer (FRET) excitation of NR on U937 monocytic cells by means of 2-photon excitation confocal laser scanning microscopy (CLSM). STUDY DESIGN: Untreated and 7KC-treated U937 cells were stained with NR and analyzed by flow cytometry and CLSM. 3D sequences of images were obtained by spectral analysis in a 2-photon excitation CLSM and analyzed by the factor analysis of medical image sequences (FAMIS) algorithm, which provides factor curves and images. Factor images are the result of the FAMIS image processing method, which handles emission spectra. In FRET analysis, preparations are screened at selected UV wavelengths to avoid emission of NR in the absence of 7KC. RESULTS: During 7KC-induced cell death,flow cytometry and CLSM revealed a modification of the cellular lipid content. Factor images show FRET occurrence and subsequent colocalization of 7KC and NR. CONCLUSION: This investigation established the utility of 2-photon excitation CLSM to assess colocalization of 7KC with NR by FRET and to identify and distinguish polar and neutral lipids stained by NR that accumulate from the effect of 7KC.     

Importance of canopy structure on photosynthesis in single- and multi-species assemblages of marine macroalgae

Plant communities utilize available irradiance with different efficiency depending not only on their photosynthetic characteristics but also on the canopy structure and density. The importance of canopy structure are well studied in terrestrial plant communities but poorly studied in aquatic plant communities. The objective of this study was to evaluate macroalgal community photosynthesis in artificial constructed communities of one to four species with different morphologies along a range of leaf (i.e.=thallus) area densities. In a laboratory set-up we measured net photosynthesis and dark respiration in constructed assemblages of macroalgae, excluding effects other than photosynthesis of individual tissue and distribution of photons in the canopy from influencing metabolism. We hypothezised that 1) canopy structure determines the actual rates of photosynthesis relative to the optimal rates and 2) multi-species communities attain higher maximum photosynthetic rates than single species communities. We found that differences in canopy structure outweighed large differences in tissue photosynthesis resulting in relatively similar maximum community photosynthetic rates among the different single and multi-species assemblages (20.1–40.5 μmol O₂ m⁻² s⁻¹). Canopy structure influenced community photosynthesis both at low and high leaf area densities because it determines the ability of macroalgae to use the photosynthetic potential of their individual tissues. Due to an averaging effect the photosynthetic rate at high leaf area density was more similar among multi-species community than among single-species communities. Multi-species communities had, on average, a slightly higher photosynthetic production than expected from photosynthesis of single species communities. Moreover multi-species communities were capable of exposing new tissue to irradiance up to high densities thereby avoiding a decrease in net photosynthesis. This finding suggests that multi-species communities may be able to maintain higher biomass per unit ground area than single-species communities.     

Mycobacterium tuberculosis recall antigens suppress HIV-1 replication in anergic donor cells via CD8+ T cell expansion and increased IL-10 levels

Mycobacterium tuberculosis (MTb) is the leading cause of death in the setting of AIDS. MTb enhances the pathogenicity and accelerates the course of HIV disease and, furthermore, infection with HIV-1 increases the risk of reactivation or reinfection with MTb. In this study, we show that host-specific recall responses to one pathogen, MTb, has a direct effect upon the regulation of a second pathogen, HIV-1. Using cells from immunocompetent former tuberculosis (TB) patients who displayed either a persistently positive (responsive) or negative (anergic), delayed-type hypersensitivity (DTH) reaction to intradermal injection of purified protein derivative (PPD), we investigated the effect of recall Ags to MTb upon the replication of HIV-1 primary isolates in vitro. We show that HIV-1 replication of a T cell-tropic isolate was significantly impaired in MTb-stimulated PBMC from PPD-anergic donors. Furthermore, these donors displayed a significant increase in CD8(+) T cells and IL-10 levels and lower levels of IL-2 and TNF-alpha relative to PPD-responsive donors in response to PPD stimulation. Strikingly, CD8(+) T cell depletion and blocking of IL-10 significantly increased HIV-1 replication in these PPD-anergic donors, indicating that an immunosuppressive response to MTb recall Ags inhibits HIV-1 replication in PPD-anergic individuals. Therefore, immunotherapeutic approaches aimed at recapitulating Ag-specific MTb anergy in vivo could result in novel and effective approaches to inhibit HIV-1 disease progression in MTb/HIV-1 coinfection.     

Uterine Msx-1 and Wnt4 signaling becomes aberrant in mice with the loss of leukemia inhibitory factor or Hoxa-10: evidence for a novel cytokine-homeobox-Wnt signaling in implantation

Successful implantation absolutely depends on the reciprocal interaction between the implantation-competent blastocyst and the receptive uterus. Expression and gene targeting studies have shown that leukemia inhibitory factor (LIF), a cytokine of the IL-6 family, and Hoxa-10, an abdominalB-like homeobox gene, are crucial to implantation and decidualization in mice. Using these mutant mice, we sought to determine the importance of Msx-1 (another homeobox gene formerly known as Hox-7.1) and of Wnt4 (a ligand of the Wnt family) signaling in implantation because of their reported functions during development. We observed that Msx-1, Wnt4, and a Wnt antagonist sFRP4 are differentially expressed in the mouse uterus during the periimplantation period, suggesting their role in implantation. In addition, we observed an aberrant uterine expression of Msx-1 and sFRP4 in Lif mutant mice, and of Wnt4 and sFRP4 in Hoxa-10 mutant mice, further reinforcing the importance of these signaling pathways in implantation. Collectively, the present results provide evidence for a novel cytokine-homeotic-Wnt signaling network in implantation.     

Has the kouprey (Bos sauveli Urbain, 1937) been domesticated in Cambodia?

The kouprey (Bos sauveli Urbain, 1937) is a very rare bovid species of Cambodia, which may be extinct in the wild, as no living specimen has been observed for a long time. Here, we describe a complete taxidermy mount, which presents astonishing morphological similarities with the kouprey. The animal was mounted in 1871 at the National Museum of Natural History in Paris, where it was referenced as No. 1871-576. It was deposited at the Natural History Museum of Bourges, France, in 1931, where it is still conserved today. To clarify the taxonomic status of the specimen of Bourges, DNA was extracted from a piece of bone taken on the mandible, and two different fragments of the mitochondrial cytochrome b gene were independently amplified and sequenced. The phylogenetic analyses show that the specimen of Bourges is robustly associated with the holotype of the kouprey, and that both are related to other wild species of Bos found in Indochina, i.e., banteng (B. javanicus) and gaur (B. frontalis). Because of doubts for sexing the animal, we applied a molecular test based on the PCR amplification of a DNA fragment specific to the Y chromosome. The results indicate that the specimen of Bourges is a male. The comparisons with male kouprey previously described in the literature reveal important differences concerning the body size, general coloration and horns. As these differences involve phenotypic traits that are strongly selected in case of domestication, we suggest that the specimen of Bourges was a domestic ox. This implies therefore that the kouprey may have been domesticated in Cambodia, and that several extant local races may be directly related to the kouprey.     

Purification of Potato Leaf Plasma Membrane Protein pp34, a Protein Phosphorylated in Response to Oligogalacturonide Signals for Defense and Development

A potato (Solanum tuberosum L.) plasma membrane protein called pp34, the only known example of a plasma membrane protein that is phosphorylated specifically in response to defined Oligogalacturonide signals in plants, has been purified to apparent homogeneity. Polyclonal antibodies raised in rabbits against the purified pp34 protein immunoprecipitated a single thiophosphorylated protein species from potato plasma membranes, as analyzed by two-dimensional denaturing electrophoresis and fluorography. The pp34 antibodies also recognized a single protein in tomato (Lycopersicon esculentum L.) membranes that is thiophosphorylated in response to Oligogalacturonide elicitors, as demonstrated by western blotting and specific immunoprecipitation. These experiments confirm the identity of the tomato membrane protein as a pp34 homolog and establish the high monospecificity of the pp34 antibodies. This will permit further investigation of the role of protein phosphorylation in oligouronide signaling for defensive genes in potato and tomato plants.