Although several surgical techniques for midclavicular fractures have been reported, Knowles pinning has rarely been compared with plating. The purpose of this study is to compare the clinical results of these two alternative techniques. There were 88 patients with midclavicular fractures surgically treated with either a Knowles pin or a plate. All patients were followed up for 12 months with a shoulder score evaluation. The Knowles pin group included 56 patients, with an average age of 40.1 years. The plate group included 32 patients, with an average age of 38.2 years. Both groups were similar in injury mechanism and fracture types (all p values>0.5). Plating has a significantly longer operation time, larger wound incision, higher pain level, more analgesic use, more complications and more symptomatic hardware (all p value<0.05). The shoulder score, union rate and healing time are not significantly different between the two groups (all p values>0.2). In conclusion, if the surgery of mid-third clavicular fractures is indicated, fixation with a Knowles pin has more advantages than plate fixation.

Limited knowledge of how plants regulate their growth and metabolism in response to drought and reduced soil water potential has impeded efforts to improve stress tolerance. Increased expression of the membrane-associated protein At14a-like1 (AFL1) led to increased growth and accumulation of the osmoprotective solute proline without negative effects on unstressed plants. Conversely, inducible RNA-interference suppression of AFL1 decreased growth and proline accumulation during low water potential while having no effect on unstressed plants. AFL1 overexpression lines had reduced expression of many stress-responsive genes, suggesting AFL1 may promote growth in part by suppression of negative regulatory genes. AFL1 interacted with the endomembrane proteins protein disulfide isomerase 5 (PDI5) and NAI2, with the PDI5 interaction being particularly increased by stress. PDI5 and NAI2 are negative regulatory factors, as pdi5, nai2, and pdi5-2nai2-3 mutants had increased growth and proline accumulation at low water potential. AFL1 also interacted with Adaptor protein2-2A (AP2-2A), which is part of a complex that recruits cargo proteins and promotes assembly of clathrin-coated vesicles. AFL1 colocalization with clathrin light chain along the plasma membrane, together with predictions of AFL1 structure, were consistent with a role in vesicle formation or trafficking. Fractionation experiments indicated that AFL1 is a peripheral membrane protein associated with both plasma membrane and endomembranes. These data identify classes of proteins (AFL1, PDI5, and NAI2) not previously known to be involved in drought signaling. AFL1-predicted structure, protein interactions, and localization all indicate its involvement in previously uncharacterized membrane-associated drought sensing or signaling mechanisms.E ven relatively mild drought that causes reduced soil water potential (ψ w ) can result in dramatically reduced plant growth and agricultural productivity. Physiological analyses have shown that plant growth is actively down-regulated during drought and is not limited by carbon supply (1-3). Reductions of growth help ensure survival by conserving water but can be undesirable for agriculture, as plant productivity is reduced more than need be if growth were less sensitive to changes in water status (3). Also, specific metabolic pathways, such as proline metabolism, are stress regulated and contribute to drought tolerance.The sensing and signaling mechanisms controlling growth and metabolic responses to drought remain unclear. Many hypotheses of how plants sense water loss center on detection of mechanical stimuli generated by loss of turgor and cell shrinkage. This includes changes in membrane shape or disruption of cell wall-cell membrane connections possibly detected by proteins, such as mechanosensitive channels or receptor-like kinases that bind cell wall components (4-9). Proteins that induce or detect membrane curvature are known in mammalian cells (10) but have been little considered in plants. Also, in analogy to mammal...

This paper describes an automated analytical system for the examination of protein primary structure in which (i) the target protein is first purified by immunoaffinity chromatography, (ii) subsequent chromatographic and chemical reaction steps in the sequencing process are directly coupled, (iii) buffer exchange between these unit operations is achieved while the protein is absorbed on a mixed bed of strong ion exchange sorbents, (iv) proteolysis occurs in an immobilized trypsin column having a 10-1000 fold-excess of enzyme, (v) the tryptic digest is directly transferred to a perfusion dilute capture column where it is concentrated and rapidly desalted, and (vi) peptides eluted from the dilute capture column and analytical microbore and capillary perfusion reversed-phase chromatography columns are analyzed by either single-stage mass spectrometry (MS) or tandem MS/MS. Protein structure variants were easily recognized, and in the case of hemoglobin (Hb) S, the site of variation from Hb A0 was verified.

An efficient system for the fast and efficient purification of transglutaminase from recombinant Streptomyces platensis and expressed in Streptomyces lividans 25-2 is described. Because the purification procedure of this system is flexible, culture broth from laboratory (20 l) and pilot-plant (130 l) fermentations were used to purify the enzyme to electrophoretic homogeneity with high purity (90-95%) and yield (61-77%) within 1 or 2 days.

Recombinant Streptomyces platensis transglutaminase (MtgA) produced by the Streptomyces lividans transformant 25-2 was purified by ammonium sulfate fractionation, followed by CM-Sepharose CL-6B fast flow, and blue-Sepharose fast flow chromatography. The purification factor was approximately 33.2-fold, and the yield was 65%. The molecular weight of the purified recombinant MtgA was 40.0 KDa as estimated by SDS-PAGE. The optimal pH and the temperature for the enzyme activity were 6.0 and 55 degrees C, respectively, and the enzyme was stable at pH 5.0-6.0 and at temperature 45-55 degrees C. Enzyme activity was not affected by Ca(2+), Li(+), Mn(2+), Na(+), Fe(3+), K(+), Mg(2+), Al(3+), Ba(2+), Co(2+), EDTA, or IAA but was inhibited by Fe(2+), Pb(2+), Zn(2+), Cu(2+), Hg(2+), PCMB, NEM, and PMSF. Optimization of the fermentation medium resulted in a twofold increase of recombinant MtgA activity in both flasks (5.78 U/ml) and 5-l fermenters (5.39 U/ml). Large-scale productions of the recombinant MtgA in a 30-l air-lift fermenter and a 250-l stirred-tank fermenter were fulfilled with maximal activities of 5.36 and 2.54 U/ml, respectively.