Identification of a novel centrosome/microtubule-associated coiled-coil protein involved in cell-cycle progression and spindle organization

Here we describe the identification of a novel vertebrate-specific centrosome/spindle pole-associated protein (CSPP) involved in cell-cycle regulation. The protein is predicted to have a tripartite domain structure, where the N- and C-terminal domains are linked through a coiled-coil mid-domain. Experimental analysis of the identified domains revealed that spindle association is dependent on the N-terminal and the coiled-coil mid domain. The expression of CSPP at the mRNA level was detected in all tested cell lines and in testis tissue. Ectopic expression of CSPP in HEK293T cells blocked cell-cycle progression in early G1 phase and in mitosis in a dose-dependent manner. Interestingly, mitosis-arrested cells contained aberrant spindles and showed impairment of chromosome congression. Inhibition of CSPP gene expression by small interfering RNAs induced cell-cycle arrest/delay in S phase. This phenotype was characterized by elevated levels of cyclin A, decreased levels of cyclin E and hyperphosphorylation of the S-phase checkpoint kinase Chk1. The activation of Chk1 may indicate a replication stress response due to an inappropriate G1/S-phase transition. Taken together, we demonstrate that CSPP is associated with centrosomes and microtubules and may play a role in the regulation of G(1)/S-phase progression and spindle assembly.     

Medical students' perceptions of resident teaching: have duty hours regulations had an impact?

OBJECTIVE: This study describes medical students' perceptions about resident teaching on a surgery clerkship and examines student perceptions before and after the implementation of duty hours regulations (DHR). SUMMARY BACKGROUND DATA: There has been much discussion about the impact of DHR on surgical education. One area that merits evaluation is the effect that DHR have had on student education. Learners perceive the clinical teacher role as comprised of 4 roles: teacher, person, physician, and supervisor. This model served as the basis for examining resident teaching before and after DHR. METHODS: Students completed end-of-rotation evaluations about residents' teaching effectiveness, amount of feedback, and quality of interactions. Student comments were compiled into individual resident reports, and reports were collected from pre- (2002-2003) and post- (2003-2004) DHR. A coding scheme was developed to describe resident performance in 4 roles: teacher, person, physician, and supervisor. Three coders independently reviewed 124 resident reports maintaining an interrater agreement of 80%. Analyses of variance were conducted to compare data from pre- and post-DHR. RESULTS: After implementation of DHR, there were significantly more negative comments (P = 0.005), including comments about residents as supervisor (P = 0.001), teacher (P = 0.027), and teaching activities (P = 0.001). Positive comments about bedside teaching decreased (P = 0.007). Although total positive comments about resident as person increased (P = 0.01), total negative comments about resident as person also increased (P = 0.02). CONCLUSIONS: Findings of this study indicate that DHR have had a negative impact on medical students' perceptions of resident teaching. Surgical educators must develop programs that address resident teaching skills in a different environment.     

Feeding, artificial sucking habits, and malocclusions in 3-year-old girls in different regions of the world

PURPOSE: The way babies and young children are reared is important to their health and development. Extensive breast-feeding has also been shown to reduce the development of artificial sucking habits like digit or pacifier-sucking. The aim of this study was to determine feeding methods, artificial sucking habits, and the presence of malocclusions in 3-year-old girls living in different regions of the world. METHODS: Children from the following countries were involved in the present study: (1) Brazil (Porto Alegre); (2) Japan (Niigata); (3) Mexico (Mexico City); (4) Norway (Oslo); (5) Sweden (Falk?ping); (6) Turkey (Istanbul); (7) and the United States (Iowa City, Iowa). During the interview and examination, the following variables were evaluated and registered: (1) breastfeeding and bottle-feeding; (2) duration and frequency; (3) sucking habits; (4) posterior and anterior crossbites; and (5) other malocclusions/normal occlusion. RESULTS: The prevalence of breast-feeding was very high in all groups, ranging between 78% and 98%. The prevalence of bottle-feeding in the different areas was also high. Except for Iowa City, the prevalence of digit-sucking was relatively low. Pacifier-sucking is fairly popular in most areas, with the exception of Niigata. The prevalence of normal occlusion in different cities ranged from 38% to 98%. CONCLUSIONS: There are considerable differences in feeding, as well as artificial sucking habits, in different areas of the world and at different periods.     

A signal sequence trap based on cell enrichment using anti-CD19 antibody coated magnetic beads

Precursors of many secreted and cell surface proteins contain NH2-terminal signal sequences that lead proteins into the endoplasmic reticulum and to the cell surface. Methods have been developed to clone and identify such proteins by trapping their NH2-terminal signal sequences, so called signal sequence traps. In this study we present an alternative and simplified signal sequence trap based on the combination of a novel vector construct expressing a cDNA library in fusion with a CD19 reporter gene, transfection in mammalian cells and selection of cells expressing trapped signal sequences using magnetic beads. Using this method we have isolated several known and novel factors with signal peptides.     

Neuropeptides in the duodenal mucosa of chronic alcoholic heavy drinkers

Previous studies have shown that patients with chronic alcohol ingestion may show a variety of morphological and functional alterations in the small intestine. In this study, we have focused on the neuroendocrine system in the duodenal mucosa in chronic alcoholics; an area little studied. Twenty-three defined chronic alcoholics admitted to the hospital for detoxification underwent clinical examination, followed by upper gastrointestinal endoscopy and blood tests on average 4 days after the most recent alcohol intake. Biopsy specimens were taken from the distal part of the descending duodenum for both immunohistochemical and routine histological examination. The control group consisted of 25 patients referred for upper endoscopy mainly because of dyspepsia (ulcer, reflux type), but who were otherwise healthy. A normal carbohydrate-deficient transferrin and a history of low alcohol consumption (<40 g/week) were required for inclusion in the control group. The tissue specimens were studied using antisera for the following neuropeptides: cholecystokinin, galanin, gastric inhibitory peptide (GIP), glucagon, motilin, neuropeptide Y, pituitary adenylyl cyclase activating peptide, secretin, serotonin, somatostatin, substance P, vasoactive intestinal polypeptide and protein gene product, as a general marker for neurones and cells of the diffuse neuroendocrine system. The density of nerve fibres was evaluated semi-quantitatively and the number of endocrine cells per unit length of mucosa was counted in sections cut perpendicularly to the mucosal surface. All the different peptidergic nerve fibres in the alcohol group showed higher densities than the corresponding controls. However, this was not a statistically significant difference. A slightly significant increase (P = 0.02) in the numbers of glucagon and GIP cells was seen in the alcohol group. Gastrointestinal symptoms were frequently present (87%) in chronic alcoholics. We suggest that chronic alcohol consumption in man may have a general effect on the peptidergic nerve system and some endocrine cell types in the duodenal mucosa.