Auxin-binding proteins without KDEL sequence in the moss Funaria hygrometrica

Whereas the important plant growth regulator auxin has multiple effects in flowering plants, it induces a specific cell differentiation step in the filamentous moss protonema. Here, we analyse the presence of classical auxin-binding protein (ABP1) homologues in the moss Funaria hygrometrica. Microsomal membranes isolated from protonemata of F. hygrometrica have specific indole acetic acid-binding sites, estimated to be about 3–5 pmol/mg protein with an apparent dissociation constant (Kd) between 3 and 5 μM. Western analyses with anti-ABP1 antiserum detected the canonical endoplasmic reticulum (ER)-localised 22–24 kDa ABP1 in Zea mays, but not in F. hygrometrica. Instead, polypeptides of 31–33 and 46 kDa were labelled in the moss as well as in maize. In F. hygrometrica these proteins were found exclusively in microsomal membrane fractions and were confirmed as ABPs by photo-affinity labelling with 5-azido-[7-3H]-indole-3-acetic acid. Unlike the classical corn ABP1, these moss ABPs did not contain the KDEL ER retention sequence. Consistently, the fully sequenced genome of the moss Physcomitrella patens, a close relative of F. hygrometrica, encodes an ABP1-homologue without KDEL sequence. Our study suggests the presence of putative ABPs in F. hygrometrica that share immunological epitopes with ABP1 and bind auxin but are different from the classical corn ABP1

Cemented hemi-arthroplasty for unstable intertrochanteric fractures in the elderly: a retrospective study

Background: Unstable intertrochanteric fractures belonging to 31A2 and A3 varieties are difficult challenges for orthopaedic surgeons, particularly in the elderly patients. Osteosynthesis by dynamic hip screw or proximal femoral nail are often plagued by complications like screw cut-out, excessive collapse and fixation failures due to osteoporotic bones. Because of these complications, patients are often kept confined to the bed which may increase the risks of pressure sores, venous thrombosis and pulmonary infections. So, it is desirable to mobilize these elderly patients as quickly as possible following surgery. In recent decades, primary hemi-arthroplasty has emerged as a valuable treatment option for mobilizing these patients early.Methods: We present our retrospective study on 27 patients above 60 years of age, who were managed with cemented bipolar hemi-arthroplasty after sustaining unstable intertrochanteric fractures. All the patients were operated through modified Hardinge approach. The fractured fragments were secured by stainless steel wiring, particularly the greater trochanter, wherever necessary.Results: Twenty-three patients (85%) were able to stand up with walkers by third post-operative day and were able to walk by fifth post-operative day. There was one case of hip dislocation; it was reduced under anaesthesia. No other complications were encountered at an average follow-up of 3.1 years. The Harris hip score was ‘good’ or ‘excellent’ in more than 60% patients.Conclusions: Cemented hemi-arthroplasty appears to be a reliable treatment method for unstable intertrochanteric fractures in the elderly and it allows early weight-bearing and rehabilitation in most patients following surgery.

Tetralogy of Fallot with rheumatic mitral stenosis: A case report

<p>Abstract</p> <p>Introduction</p> <p>Rheumatic and congenital heart diseases account for the majority of hospital admissions for cardiac patients in India. Tetralogy of Fallot is the most common congenital heart disease with survival to adulthood. Infective endocarditis accounts for 4% of admissions to a specialized unit for adult patients with a congenital heart lesion. This report is unique in that a severe stenotic lesion of the mitral valve, probably of rheumatic aetiology, was noted in an adult male with Tetralogy of Fallot.</p> <p>Case presentation</p> <p>An unusual association of rheumatic mitral stenosis in an adult Indian male patient aged 35 years with Tetralogy of Fallot and subacute bacterial endocarditis of the aortic valve is presented.</p> <p>Conclusion</p> <p>In this case report the diagnostic implications, hemodynamic and therapeutic consequences of mitral stenosis in Tetralogy of Fallot are discussed. In addition, the morbidity and mortality of infective endocarditis in adult patients with congenital heart disease are summarized. The risk of a coincident rheumatic process in patients with congenital heart disease is highlighted and the need for careful attention to this possibility during primary and follow-up evaluation of such patients emphasized.</p

Arabidopsis COP1 shapes the temporal pattern of CO accumulation conferring a photoperiodic flowering response

The transcriptional regulator CONSTANS (CO) promotes flowering of Arabidopsis under long summer days (LDs) but not under short winter days (SDs). Post-translational regulation of CO is crucial for this response by stabilizing the protein at the end of a LD, whereas promoting its degradation throughout the night under LD and SD. We show that mutations in CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), a component of a ubiquitin ligase, cause extreme early flowering under SDs, and that this is largely dependent on CO activity. Furthermore, transcription of the CO target gene FT is increased in cop1 mutants and decreased in plants overexpressing COP1 in phloem companion cells. COP1 and CO interact in vivo and in vitro through the C-terminal region of CO. COP1 promotes CO degradation mainly in the dark, so that in cop1 mutants CO protein but not CO mRNA abundance is dramatically increased during the night. However, in the morning CO degradation occurs independently of COP1 by a phytochrome B-dependent mechanism. Thus, COP1 contributes to day length perception by reducing the abundance of CO during the night and thereby delaying flowering under SDs

Improved methods to detect GTP-binding proteins from plants

Improved methods are described for the detection of G1P-binding proteins (G-proteins) in the protonema of mossFunaria hygrometrica and coleoptiles of corn (Zea mays) and sorghum(Sorghum vulgare). We optimized conditions for the transfer of proteins to nitrocellulose, production of high titer polyclonal anti-Ga (common) antibodies and finally the detection of G-proteins by amplification. In addition to the &#945; -subunit of heterotrimeric G-proteins (Mr 41-43 kDa), a small molecular weight class (&lt; 30 kDa) was also detected by anti-Ga (common) antibodies. An easy, reliable and efficient filter assay is also described to quantify the toxin catalyzed ADP-ribosylation. The apparent Km of the NAD has been determined to be approximately 1.5&#181; M for the microsomal fraction of moss. Inclusion of G1P stimulated ADP-ribosylation by 2-27-fold. One to three polypeptides representing the &#945; -subunit of heterotrimeric G-proteins of (Mr 37-43 kDa) were ADP-ribosylated in all three plants. The anti-G&#946; (C-terminus) antibody cross-reacted strongly with 39 and 34 kDa polypeptide in moss and corn respectively. By employing improved methods two classes of G-proteins have been shown to be present in three plant species