S100A8 chemotactic protein is abundantly increased, but only a minor contributor to LPS-induced, steroid resistant neutrophilic lung inflammation in vivo

Neutrophilic lung inflammation is an essential component of host defense against diverse eukaryotic and prokaryotic pathogens, but in chronic inflammatory lung diseases, such as chronic obstructive lung disease (COPD), severe asthma, cystic fibrosis, and bronchiolitis, it may damage the host. Glucocorticosteroids are widely used in these conditions and in their infectious exacerbations; however, the clinical efficacy of steroids is disputed. In this study, we used a proteomic approach to identify molecules contributing to neutrophilic inflammation induced by transnasal administration of lipopolysaccharide (LPS) that were also resistant to the potent glucocorticosteroid dexamethasone (Dex). We confirmed that Dex was biologically active at both the transcript (suppression of GM-CSF and TNFalpha transcripts) and protein levels (induction of lipocortin) and used 2D-PAGE/MALDI-TOF to generate global expression profiles, identifying six LPS-induced proteins that were Dex resistant. Of these, S100A8, a candidate neutrophil chemotactic factor, was profiled in detail. Steroid refractory S100A8 expression was highly abundant, transcriptionally regulated, secreted into lung lavage fluid and immunohistochemically localized to tissue infiltrating neutrophils. However, in marked contrast to other vascular beds, neutralizing antibodies to S100A8 had only a weak anti-neutrophil recruitment effect and antibodies against the related S100A9 were ineffective. These data highlight the need for extensive in vivo profiling of proteomically identified candidate molecules and demonstrates that S100A8, despite its abundance, resistance to steroids and known chemotactic activity, is unlikely to be an important determinant of LPS-induced neutrophilic lung inflammation in vivo.

Evidence of altered lumbopelvic muscle recruitment in the presence of sacroiliac joint pain

Study Design. Cross-sectional study of electromyographic onsets of trunk and hip muscles in subjects with a clinical diagnosis of sacroiliac joint pain and matched control subjects. Objectives. To determine whether muscle activation of the supporting leg was different between control subjects and subjects with sacroiliac joint pain during hip flexion in standing. Background. Activation of the trunk and gluteal muscles stabilize the pelvis for load transference; however, the temporal pattern of muscle activation and the effect of pelvic pain on temporal parameters has not been investigated. Methods. Fourteen men with a clinical diagnosis of sacroiliac joint pain and healthy age-matched control subjects were studied. Surface electromyographic activity was recorded from seven trunk and hip muscles of the supporting leg during hip flexion in standing. Onset of muscle activity relative to initiation of the task was compared between groups and between limbs. Results. The onset of obliquus internus abdominis (OI) and multifidus occurred before initiation of weight transfer in the control subjects. the onset of obliquus internus abdominis, multifidus, and gluteus maximus was delayed on the symptomatic side in subjects with sacroiliac joint pain compared with control subjects, and the onset of biceps femoris electromyographic activity was earlier. IN addition, electromyographic onsets were different between the symptomatic and asymptomatic sides in subjects with sacroiliac joint pain. Conclusions. The delayed onset of obliquus internus abdominis, multifidus, and gluteus maximus electromyographic activity of the supporting leg during hip flexion, in subjects with sacroiliac joint pain. suggests an alteration in the strategy for lumbopelvic stabilization that may disrupt load transference through the pelvis.

Abdominal muscle recruitment during a range of voluntary exercises

Various exercises are used to retrain the abdominal muscles in the management of low back pain and other musculoskeletal disorders. However. few studies have directly investigated the activity of all the abdominal muscles or the recruitment of regions of the abdominal muscles during these manoeuvres. This study examined the activity of different regions of transversus abdominis (TrA), obliquus internus (OI) and externus abdominis (OE), and rectus abdominis (RA), and movement of lumbar spine, pelvis and abdomen during inward movement of the lower abdominal wall, abdominal bracing, pelvic tilting, and inward movement of the lower and upper abdominal wall. Inward movement of the lower abdominal wall in supine produced greater activity of TrA compared to OI. OE and RA. During posterior pelvic tilting. middle OI was most active and with abdominal bracing. OE was predominately recruited. Regions of TrA were recruited differentially and in inverse relationship between lumbopelvic motion and TrA electromyography (EMG) was found. This study indicates that inward movement of the abdominal wall in supine produces the most independent activity of TrA relative to the other abdominal muscle, recruitment varies between regions of TrA, and observation of abdominal and lumbopelvic motion may assist in evalation of exercise performance. (c) 2004 Elsevier Ltd. All rights reserved.

Cdk1/Erk2- and Plk1-dependent phosphorylation of a centrosome protein, Cep55, is required for its recruitment to midbody and cytokinesis

Centrosomes in mammalian cells have recently been implicated in cytokinesis; however, their role in this process is poorly defined. Here, we describe a human coiled-coil protein, Cep55 (centrosome protein 55 kDa), that localizes to the mother centriole during interphase. Despite its association with gamma-TuRC anchoring proteins CG-NAP and Kendrin, Cep55 is not required for microtubule nucleation. Upon mitotic entry, centrosome dissociation of Cep55 is triggered by Erk2/Cdk1-dependent phosphorylation at S425 and S428. Furthermore, Cep55 locates to the midbody and plays a role in cytokinesis, as its depletion by siRNA results in failure of this process. S425/428 phosphorylation is required for interaction with Plk1, enabling phosphorylation of Cep55 at S436. Cells expressing phosphorylation-deficient mutant forms of Cep55 undergo cytokinesis failure. These results highlight the centrosome as a site to organize phosphorylation of Cep55, enabling it to relocate to the midbody to function in mitotic exit and cytokinesis.

Leg muscle recruitment in highly trained cyclists

In this study, we examined patterns of leg muscle recruitment and co-activation, and the relationship between muscle recruitment and cadence, in highly trained cyclists. Electromyographic (EMG) activity of the tibialis anterior, tibialis posterior, peroneus longus, gastrocnemius lateralis and soleus was recorded using intramuscular electrodes, at individual preferred cadence, 57.5, 77.5 and 92.5 rev.min(-1). The influence of electrode type and location on recorded EMG was also investigated using surface and dual intramuscular recordings. Muscle recruitment patterns varied from those previously reported, but there was little variation in muscle recruitment between these highly trained cyclists. The tibialis posterior, peroneus longus and soleus were recruited in a single, short burst of activity during the downstroke. The tibialis anterior and gastrocnemius lateralis were recruited in a biphasic and alternating manner. Contrary to existing hypotheses, our results indicate little co-activation between the tibialis posterior and peroneus longus. Peak EMG amplitude increased linearly with cadence and did not decrease at individual preferred cadence. There was little variation in patterns of muscle recruitment or co-activation with changes in cadence. Intramuscular electrode location had little influence on recorded EMG. There were significant differences between surface and intramuscular recordings from the tibialis anterior and gastrocnemius lateralis, which may explain differences between our findings and those of previous studies.

Visualisation of macropinosome maturation by the recruitment of sorting nexins

We report that phosphoinositol-binding sorting nexin 5 ( SNX5) associates with newly formed macropinosomes induced by EGF stimulation. We used the recruitment of GFP-SNX5 to macropinosomes to track their maturation. Initially, GFP-SNX5 is sequestered to discrete subdomains of the macropinosome; these subdomains are subsequently incorporated into highly dynamic, often branched, tubular structures. Time-lapse videomicroscopy revealed the highly dynamic extension of SNX5-labelled tubules and their departure from the macropinosome body to follow predefined paths towards the perinuclear region of the cell, before fusing with early endosomal acceptor membranes. The extension and departure of these tubular structures occurs rapidly over 5-10 minutes and is dependent upon intact microtubules. As the tubular structures depart from the macropinosome there is a reduction in the surface area and an increase in tension of the limiting membrane of the macropinosome. In addition to the recruitment of SNX5 to the macropinosome, Rab5, SNX1 and EEA1 are also recruited by newly formed macropinosomes, followed by the accumulation of Rab7. SNX5 forms heterodimers with SNX1 and this interaction is required for endosome association of SNX5. We propose that the departure of SNX5-positive tubules represents a rapid mechanism of recycling components from macropinosomes thereby promoting their maturation into Rab7-positive structures. Collectively these findings provide a detailed real-time characterisation of the maturation process of the macropinocytic endosome.

The rural medicine rotation: Increasing rural recruitment through quality undergraduate rural experiences

TITLE: The Rural Medicine Rotation: Increasing Rural Recruitment through Quality Undergraduate Rural Experiences Eley Diann, University of Queensland, School of Medicine, Rural Clinical Division, Toowoomba 4350, Queensland Australia Baker Peter, University of Queensland, School of Medicine Rural, Clinical Division, Toowoomba 4350, Queensland Australia Chater Bruce, University of Queensland, Chair, Clinical School Management Committee, School of Medicine Rural Clinical Division, Queensland Australia CONTEXT: While rural background and rural exposure during medical training increases the likelihood of rural recruitment (Wilkinson, 2003), the quality and content of that exposure is the key to altering undergraduatesâ?? perceptions of rural practice. The Rural Clinical Division at University of Queensland (UQ) runs the Rural Medicine Rotation (RMR) within the School of Medicine. The RMR is one of five eight week clinical rotations in Year three and is compulsory for all students. The RMR provides the opportunity to learn from a wide range of health professionals and clinical exposure is not restricted to general practice but also includes remote area nursing, Indigenous health care, allied health professionals and medical specialists. Week 1 involves preparation for their rural placement with workshops and seminars and Week 8 consolidates their placement and includes case and project presentations and a summative assessment. Weeks 2-7 are spent living and working as part of the health team in different rural communities. SETTING: Rural communities in and around Queensland including locations such as Arnham Land, Thursday Island, Mt. Isa and Alice Springs METHOD: All aspects of the RMR are evaluated with surveys using both qualitative and quantitative free response questions, completed by all students at the end of the Week 8. RESULTS: Overall the RMR is evaluated highly and narratives offered by students show that the RMR provides a positive rural experience. The overall impact of the RMR for students in 2004 ranked 3.45 on a scale of 1 to 4 (1 = lowest and 4 = highest), and is exemplified by the following quote; â??I enjoyed my placement so much I am now considering rural medicine something I definitely had not considered beforeâ??. OUTCOME: The positive impact of the RMR on studentâ??s perceptions of rural medicine is encouraging and can help achieve the overall aim of increasing recruitment of the rural workforce in Australia.