TGF β-induced cartilage repair is maintained but fibrosis is blocked in the presence of Smad7

Cartilage damage in osteoarthritis (OA) is considered an imbalance between catabolic and anabolic factors, favoring the catabolic side. We assessed whether adenoviral overexpression of transforming growth factor-β (TGFβ) enhanced cartilage repair and whether TGFβ-induced fibrosis was blocked by local expression of the intracellular TGFβ inhibitor Smad7. We inflicted cartilage damage by injection of interleukin-1 (IL-1) into murine knee joints. After 2 days, we injected an adenovirus encoding TGFβ. On day 4, we measured proteoglycan (PG) synthesis and content. To examine whether we could block TGFβ-induced fibrosis and stimulate cartilage repair simultaneously, we injected Ad-TGFβ and Ad-Smad7. This was performed both after IL-1-induced damage and in a model of primary OA. In addition to PG in cartilage, synovial fibrosis was measured by determining the synovial width and the number of procollagen I-expressing cells. Adenoviral overexpression of TGFβ restored the IL-1-induced reduction in PG content and increased PG synthesis. TGFβ-induced an elevation in PG content in cartilage of the OA model. TGFβ-induced synovial fibrosis was strongly diminished by simultaneous synovial overexpression of Smad7 in the synovial lining. Of great interest, overexpression of Smad7 did not reduce the repair-stimulating effect of TGFβ on cartilage. Adenoviral overexpression of TGFβ stimulated repair of IL-1- and OA-damaged cartilage. TGFβ-induced synovial fibrosis was blocked by locally inhibiting TGFβ signaling in the synovial lining by simultaneously transfecting it with an adenovirus overexpressing Smad7.     

Report on the Polycystididae (Rhabdocoela, Kalyptorhynchia) from Australia, with the Description of Twelve New Species and Six New Genera

Twelve new species of Polycystididae Graff, 1905 are described from the Australian east coast. Nine of them could not be placed in any of the existing genera, and therefore six new genera are erected. Alchoides n. gen. differs from all other polycystidid genera by the presence of a bundle of glands that opens into the distal part of the male atrium (accessory vesicle type V). Two species are included in the genus, A. alchoides n. sp. and A. dittmannae n. sp., which differ from each other in the shape of the stylet. Ametochus gehrkei n. gen n. sp. can be distinguished from other polycystidids by the presence of a prostate vesicle type III associated with a prostate stylet type III, combined with the presence of an accessory stylet type III, while a prostate stylet type II is lacking. Typical for Arrawarria inexpectata n. gen n. sp. is the combined presence of an armed cirrus with a prostate vesicle type II connected to a prostate stylet type II. Two species are assigned to the new genus Duplexostylus n. gen.: D. rowei n. sp. and D. winsori n. sp. These two species have two unique features: a prostate vesicle type IV directly connected to a prostate stylet type III and an asymmetrical septum that surrounds the proximal part of the male atrium. They can be distinguished from each other by differences in the detailed construction of their stylets. Two new species are placed in the new genus Stradorhynchus n. gen.: S. caecus n. sp. and S. terminalis n. sp. They have unpaired gonads and a prostate vesicle type IV connected to a prostate stylet type IV, which combination of characters makes them unique within the Polycystididae. The two species differ from each other in the shape and size of the stylet. Triaustrorhynchus armatus n. gen., n. sp. is the first species of the Polycystididae to have three types of stylet in the male atrium: a prostate stylet type II, a prostate stylet type III and an accessory stylet type III. Three new species could be placed in existing genera: Cincturorhynchus monaculeus n. sp., Paraustrorhynchus caligatus n. sp. and Polycystis australis n. sp. All three differ from their congeners in the form and dimensions of their stylets. All newly described species are discussed, giving their similarities and differences with resembling species. Austrorhynchus hawaiiensis Karling, 1977 is reported for the first time in Australia, and the Australian population is compared with populations from other regions. The presence of Gyratrix hermaphroditus Ehrenberg, 1831, one new species of Paulodora Marcus, 1948 and a new genus of Typhlopolycystidinae Evdonin, 1977 is mentioned.     

Tracing of labile zinc in live fish hepatocytes using FluoZin-3

Intracellular zinc levels are homeostatically regulated and although most is bound, a pool of labile Zn(II) is present in cells. We show here that the zinc probe FluoZin-3 is useful to monitor zinc fluxes during fluorescent imaging of the trout hepatic cell line D11. Nuclei and bulk cytosol appeared to lack detectable labile zinc, while the punctuate staining pattern colocalized with a lysosome-specific probe. Applying extracellular zinc alone resulted in vesicular sequestration of the metal ion. Together with Na-pyrithione a delayed and toxic rise in cellular fluorescence was triggered. When using another ionophore, 4-Br A23187, a zinc buffering effect of the vesicular pools was evident. Secondly, N-ethylmaleimide induced a homogeneous fluorescence rise, which was strongly enhanced by addition of Zn-pyrithione and disappeared after TPEN washing. This suggests the involvement of thiol residues in controlling available cytosolic zinc. Our observations have implications for the interpretation of calculated intracellular Zn²⁺ concentrations.     

Serum proteomics in amnestic mild cognitive impairment

We have explored proteins related to mild cognitive impairment (MCI). The serum proteome of 35 amnestic MCI patients and 35 cognitively healthy persons was investigated by LC MS. We identified 108 differentially expressed peptides between MCI patients and controls, belonging to 39 proteins. Eight proteins were selected for further investigation by quantitative protein measurements using a MRM assay; apolipoprotein E, carboxypeptidase N subunit 2, complement factor B (CFAB), galectin‐3 binding protein (LG3BP), lumican, serum amyloid A‐4 protein (SAA4), serum amyloid P‐component, and sex hormone binding globulin. Results of the quantitative protein measurements showed significantly decreased levels of carboxypeptidase N subunit 2, CFAB, LG3BP, SAA4, and serum amyloid P‐component in serum from amnestic MCI patients compared with cognitive healthy controls (two‐sided t‐test; p < 0.05). Apolipoprotein E and lumican showed no significant difference in protein levels, sex hormone binding globulin could not be quantified since the MRM assay did not reach the required sensitivity. A model based on the three most significantly decreased proteins (CFAB, LG3BP, and SAA4) showed a sensitivity and specificity of 73 and 66%, respectively, for the initial sample set. A small external validation set yielded 77% sensitivity and 75% specificity.     

Detecting Cheaters without Thinking: Testing the Automaticity of the Cheater Detection Module

Evolutionary psychologists have suggested that our brain is composed of evolved mechanisms. One extensively studied mechanism is the cheater detection module. This module would make people very good at detecting cheaters in a social exchange. A vast amount of research has illustrated performance facilitation on social contract selection tasks. This facilitation is attributed to the alleged automatic and isolated operation of the module (i.e., independent of general cognitive capacity). This study, using the selection task, tested the critical automaticity assumption in three experiments. Experiments 1 and 2 established that performance on social contract versions did not depend on cognitive capacity or age. Experiment 3 showed that experimentally burdening cognitive resources with a secondary task had no impact on performance on the social contract version. However, in all experiments, performance on a non-social contract version did depend on available cognitive capacity. Overall, findings validate the automatic and effortless nature of social exchange reasoning.     

Structure of an early native‐like intermediate of β2‐microglobulin amyloidogenesis

To investigate early intermediates of β2‐microglobulin (β2m) amyloidogenesis, we solved the structure of β2m containing the amyloidogenic Pro32Gly mutation by X‐ray crystallography. One nanobody (Nb24) that efficiently blocks fibril elongation was used as a chaperone to co‐crystallize the Pro32Gly β2m monomer under physiological conditions. The complex of P32G β2m with Nb24 reveals a trans peptide bond at position 32 of this amyloidogenic variant, whereas Pro32 adopts the cis conformation in the wild‐type monomer, indicating that the cis to trans isomerization at Pro32 plays a critical role in the early onset of β2m amyloid formation.