An LRR receptor kinase controls ABC transporter substrate preferences during plant growth-defense decisions.

The exporter of the auxin precursor indole-3-butyric acid (IBA), ABCG36/PDR8/PEN3, from the model plant Arabidopsis has recently been proposed to also function in the transport of the phytoalexin camalexin. Based on these bonafide substrates, it has been suggested that ABCG36 functions at the interface between growth and defense. Here, we provide evidence that ABCG36 catalyzes the direct, ATP-dependent export of camalexin across the plasma membrane. We identify the leucine-rich repeat receptor kinase, QIAN SHOU KINASE1 (QSK1), as a functional kinase that physically interacts with and phosphorylates ABCG36. Phosphorylation of ABCG36 by QSK1 unilaterally represses IBA export, allowing camalexin export by ABCG36 conferring pathogen resistance. As a consequence, phospho-dead mutants of ABCG36, as well as qsk1 and abcg36 alleles, are hypersensitive to infection with the root pathogen Fusarium oxysporum, caused by elevated fungal progression. Our findings indicate a direct regulatory circuit between a receptor kinase and an ABC transporter that functions to control transporter substrate preference during plant growth and defense balance decisions.

Detection of MCPG metabolites in horses with atypical myopathy.

Atypical myopathy (AM) in horses is caused by ingestion of seeds of the Acer species (Sapindaceae family). Methylenecyclopropylacetyl-CoA (MCPA-CoA), derived from hypoglycin A (HGA), is currently the only active toxin in Acer pseudoplatanus or Acer negundo seeds related to AM outbreaks. However, seeds or arils of various Sapindaceae (e.g., ackee, lychee, mamoncillo, longan fruit) also contain methylenecyclopropylglycine (MCPG), which is a structural analogue of HGA that can cause hypoglycaemic encephalopathy in humans. The active poison formed from MCPG is methylenecyclopropylformyl-CoA (MCPF-CoA). MCPF-CoA and MCPA-CoA strongly inhibit enzymes that participate in ß-oxidation and energy production from fat. The aim of our study was to investigate if MCPG is involved in Acer seed poisoning in horses. MCPG, as well as glycine and carnitine conjugates (MCPF-glycine, MCPF-carnitine), were quantified using high-performance liquid chromatography-tandem mass spectrometry of serum and urine from horses that had ingested Acer pseudoplatanus seeds and developed typical AM symptoms. The results were compared to those of healthy control horses. For comparison, HGA and its glycine and carnitine derivatives were also measured. Additionally, to assess the degree of enzyme inhibition of ß-oxidation, several acyl glycines and acyl carnitines were included in the analysis. In addition to HGA and the specific toxic metabolites (MCPA-carnitine and MCPA-glycine), MCPG, MCPF-glycine and MCPF-carnitine were detected in the serum and urine of affected horses. Strong inhibition of ß-oxidation was demonstrated by elevated concentrations of all acyl glycines and carnitines, but the highest correlations were observed between MCPF-carnitine and isobutyryl-carnitine (r = 0.93) as well as between MCPA- (and MCPF-) glycine and valeryl-glycine with r = 0.96 (and r = 0.87). As shown here, for biochemical analysis of atypical myopathy of horses, it is necessary to take MCPG and the corresponding metabolites into consideration.

In vivo wear rate of the Birmingham Hip Resurfacing arthroplasty. A review of 10 retrieved components.

The wear of 8 femoral and 2 acetabular components of the Birmingham Hip Resurfacing (Midland Medical Technologies Ltd, Birmingham, UK) was measured using a coordinate measuring machine (CMM5; SIP, Geneva, Switzerland). After a median in situ time of 13 months, measurements of the Birmingham Hip Resurfacing femoral heads revealed a median volumetric wear of 2.9 mm(3). In one case with a cup abduction angle of 70 degrees , a significantly higher wear of 17.8 mm(3) was found. The 2 acetabular components showed volumetric wear of 5.4 and 27.6 mm(3) after 14 and 15 months in vivo. The wear rates of the investigated femoral components were somewhat, but not significantly, higher than the previously investigated 28-mm Metasul heads (median, 0.8 mm(3); Zimmer GmbH, Winterthur, Switzerland).

Biological activity of recombinant human growth factors released from biocompatible bone implants.

The present investigation was performed to study the bioactivity of osteoinductive and osteoproliferative growth factors after release from biocompatible bone implants. Three types of porous carriers were used in this study: hydroxyapatite, alpha tricalcium phosphate, and a neutralized glass ceramic. Implants were loaded with recombinant human bone morphogenetic protein 2 (rh-BMP-2) and recombinant human basic fibroblast growth factor (rh-bFGF) in a concentration of 2 microg/150 microL PBS each. The released growth factors were then applicated into SAOS-2-cell cultures. After 3, 5, and 7 days cell differentiation was measured by the activity of alkaline phosphatase (ALP), cell proliferation by using a MTT assay as well as a cell counter. Rh-BMP-2 released during the first hour from the scaffolds led to a significant increase of the activity of ALP in the incubated SAOS-2-cell culture after 3, 5, and 7 days. However, the incubation with rh-BMP-2 released after 24 h was not found to increase the expression of ALP. The incubation of cell cultures with rh-bFGF released during the first hour led to a significant increase of cell number and of extinction in the MTT assay, whereas this increase was not observed after incubation with rh-bFGF released after 24 h. The in vitro measured biological activity of released growth factors from the surface of synthetic implants is time-depending. If prolonged osteoinductive and osteoproliferative potency of growth factors is desired, a modified application technique should be chosen to stabilize those proteins.

Serum metal ion exposure after total knee arthroplasty.

All metal implants release metal ions because of corrosion. This has been studied and debated, especially in metal-on-metal total hip arthroplasties. Total knee arthroplasty implants have large metal surface areas and therefore substantial potential for corrosion. We determined changes in serum levels of metal ions in 41 patients after cemented unconstrained total knee arthroplasty without patellar resurfacing, 18 with unilateral total knee arthroplasty (median, 66 months after surgery) and 23 patients with bilateral total knee arthroplasties (75 and 50 months after first and second surgeries, respectively). Serum concentrations of chromium, cobalt, and molybdenum were analyzed and related to the number of total knee arthroplasties and compared with those of 130 control patients without implants. The median chromium, cobalt, and molybdenum concentrations were 0.92, 3.28, and 2.55 microg/L, respectively, in the unilateral total knee arthroplasty sample and 0.98, 4.28, and 2.40 microg/L, respectively, in the bilateral total knee arthroplasty sample. We observed no difference between the serum levels in patients with unilateral and bilateral arthroplasties, but the serum levels of chromium and cobalt of both study groups were greater than those of the control group (less than 0.25 microg/L). The patients who had total knee arthroplasty had molybdenum profiles that were similar to those of the control group (median, 2.11 microg/L).

Exposure to chromium, cobalt and molybdenum from metal-on-metal total hip replacement and hip resurfacing arthroplasty.

BACKGROUND: All metal implants--and metal-on-metal bearings in particular--corrode and cause a release of metal ions. Because cobalt and chromium have been shown to be carcinogenic and mutagenic in human and animal models, systemic toxicity and cancer risk are considered to be possible disadvantages of the metal-on-metal articulation. This study was designed to investigate the serum concentration profiles of chromium, cobalt and molybdenum after implantation of a Birmingham hip resurfacing arthroplasty (BHR) and a cementless total hip replacement with a 28-mm Metasul articulation (MTHR), over the first 2 years after implantation. METHODS: We analyzed profiles of metal ion serum levels in 111 patients implanted with a BHR, in 74 patients implanted with an MTHR, and in 130 implant-free probands control subjects using atomic absorption spectrophotometry. RESULTS: Chromium and cobalt concentrations (in microg/L) of all BHR and MTHR patients differed significantly from those of control subjects (chromium: < 0.25; cobalt: 0.25). The median chromium and cobalt concentrations in BHR patients had increased to 5.1 and 4.3 microg/L 2 years after surgery. Concentrations in BHR patient exceeded those in the unilateral MTHR patients. Molybdenum serum concentrations hardly changed over time in either group and were not significantly different from the concentrations seen in the control subjects. INTERPRETATION: During the first 2 years after surgery, the Birmingham hip resurfacing arthroplasty leads to a significantly greater increase in serum chromium and cobalt levels than the 28-mm metal-on-metal MTHR. Observation of patients over a longer period will be necessary in order to evaluate any chronic adverse effects to the system due to elevated chromium and cobalt serum concentrations.

Toward intelligent nanosize bioreactors: a pH-switchable, channel-equipped, functional polymer nanocontainer.

To develop an intelligent sensor-effector functionality on the nanoscale, a pH-switchable, controlled nanoreactor based on amphiphilic copolymer membranes was built. The nanovesicles were equipped with bacterial transmembrane ompF pore proteins and the pH-sensitive enzyme acid phosphatase, resulting in a switchable substrate processing at pH 4-6.5. Ideal pH and substrate concentrations for the reaction were determined experimentally. In future, the reactor might be used for self-regulating targeted diagnostic and therapeutic applications in medicine.