DLK1 Is a Somato-Dendritic Protein Expressed in Hypothalamic Arginine-Vasopressin and Oxytocin Neurons

Delta-Like 1 Homolog, Dlk1, is a paternally imprinted gene encoding a transmembrane protein involved in the differentiation of several cell types. After birth, Dlk1 expression decreases substantially in all tissues except endocrine glands. Dlk1 deletion in mice results in pre-natal and post-natal growth deficiency, mild obesity, facial abnormalities, and abnormal skeletal development, suggesting involvement of Dlk1 in perinatal survival, normal growth and homeostasis of fat deposition. A neuroendocrine function has also been suggested for DLK1 but never characterised. To evaluate the neuroendocrine function of DLK1, we first characterised Dlk1 expression in mouse hypothalamus and then studied post-natal variations of the hypothalamic expression. Western Blot analysis of adult mouse hypothalamus protein extracts showed that Dlk1 was expressed almost exclusively as a soluble protein produced by cleavage of the extracellular domain. Immunohistochemistry showed neuronal DLK1 expression in the suprachiasmatic (SCN), supraoptic (SON), paraventricular (PVN), arcuate (ARC), dorsomedial (DMN) and lateral hypothalamic (LH) nuclei. DLK1 was expressed in the dendrites and perikarya of arginine-vasopressin neurons in PVN, SCN and SON and in oxytocin neurons in PVN and SON. These findings suggest a role for DLK1 in the post-natal development of hypothalamic functions, most notably those regulated by the arginine-vasopressin and oxytocin systems

Does mechanical massage of the abdominal wall after colectomy reduce postoperative pain and shorten the duration of ileus? Results of a randomized study

The aim of this study was to determine the effectiveness of mechanical abdominal massage on postoperative pain and ileus after colectomy. We hypothesized that parietal abdominal stimulation could counteract induced pain and postoperative ileus, through common spinal-sensitive pathways, with nociceptive visceral messages. After preoperative randomization, 25 patients (age 52 ± 5 years) underwent active mechanical massage by intermittent negative pressure on the abdominal wall resulting in aspiration (Cellu M50 device, LPG, Valence, France), and 25 patients (age 60 ± 6 years) did not receive active mechanical massage (placebo group). Massage sessions began the first day after colectomy and were performed daily until the seventh postoperative day. In the active-massage group, amplitude and frequency were used, which have been shown to be effective in reducing muscular pain, whereas in the placebo group, ineffective parameters were used. Visual analogue scale (VAS) pain scores, doses of analgesics (propacetamol), and delay between surgery and the time to first passage of flatus were assessed. Types and dosages of the anesthetic drugs and the duration of the surgical procedure did not differ between groups. From the second and third postoperative days, respectively, VAS pain scores (P \u3c 0.001) and doses of analgesics (P \u3c 0.05) were significantly lower in patients receiving active massage compared to the placebo group. Time to first passage of flatus was also significantly shorter in the active-massage group (1.8 ± 0.3 days vs. 3.6 ± 0.4 days, P \u3c 0.01). No adverse effects were observed. These results suggest that mechanical massage of the abdominal wall may decrease postoperative pain and ileus after colectomy. © 2002 The Society for Surgery of the Alimentary Tract, Inc

Electrotransformation of Saccharomyces cerevisiae

Intact yeast cell transformation is easily achieved by gene electrotransfer (GET). The procedure is fast and efficient in terms of transformants/mu g DNA. Yeast cells in exponential growth phase are washed, treated for a short period with dithiothreitol (DTT) and then mixed with the plasmid DNA in a buffer with a low conductivity. A single well defined electric pulsed is delivered. After a 1 h incubation in the growth medium without selection, transformants are obtained on a selective plate medium. After a short description of the present knowledge on the events affecting the yeast cell as a consequence of the pulsed electric field, a step-by-step protocol is reported for Saccharomyces cerevisiae

Exploring early steps in biofilm formation: set-up of an experimental system for molecular studies

Background: Bacterial biofilms are predominant in natural ecosystems and constitute a public health threat because of their outstanding resistance to antibacterial treatments and especially to antibiotics. To date, several systems have been developed to grow bacterial biofilms in order to study their phenotypes and the physiology of sessile cells. Although relevant, such systems permit analysis of various aspects of the biofilm state but often after several hours of bacterial growth.Results: Here we describe a simple and easy-to-use system for growing P. aeruginosa biofilm based on the medium adsorption onto glass wool fibers. This approach which promotes bacterial contact onto the support, makes it possible to obtain in a few minutes a large population of sessile bacteria. Using this growth system, we demonstrated the feasibility of exploring the early stages of biofilm formation by separating by electrophoresis proteins extracted directly from immobilized cells. Moreover, the involvement of protein synthesis in P. aeruginosa attachment is demonstrated.Conclusions: Our system provides sufficient sessile biomass to perform biochemical and proteomic analyses from the early incubation period, thus paving the way for the molecular analysis of the early stages of colonization that were inaccessible to date