A Cell Cycle Phosphoproteome of the Yeast Centrosome Keck, Jamie M.; Jones, Michele H.; Wong, Catherine C. L. ...
Science (American Association for the Advancement of Science),
06/2011, Letnik:
332, Številka:
6037
Journal Article
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Centrosomes organize the bipolar mitotic spindle, and centrosomal defects cause chromosome instability. Protein phosphorylation modulates centrosome function, and we provide a comprehensive map of ...phosphorylation on intact yeast centrosomes (18 proteins). Mass spectrometry was used to identify 297 phosphorylation sites on centrosomes from different cell cycle stages. We observed different modes of phosphoregulation via specific protein kinases, phosphorylation site clustering, and conserved phosphorylated residues. Mutating all eight cyclin-dependent kinase (Cdk)—directed sites within the core component, Spc42, resulted in lethality and reduced centrosomal assembly. Alternatively, mutation of one conserved Cdk site within γ-tubulin (Tub4-S360D) caused mitotic delay and aberrant anaphase spindle elongation. Our work establishes the extent and complexity of this prominent posttranslational modification in centrosome biology and provides specific examples of phosphorylation control in centrosome function.
The two municipal drinking water systems of New Orleans, LA, U.S.A. were sampled to compare the microbiology of independent systems that treat the same surface water from the Mississippi River. To ...better understand temporal trends and sources of microbiology delivered to taps, these treatment plants and distribution systems were subjected to source-to-tap sampling over four years. Both plants employ traditional treatment by chloramination, applied during or after settling, followed by filtration before distribution in a warm, low water age system. Longitudinal samples indicated microbiology to have stability both spatially and temporally, and between treatment plants and distribution systems. Disinfection had the greatest impact on microbial composition, which was further refined by filtration and influenced by distribution and premise plumbing. Actinobacteria spp. exhibited trends with treatment. In particular, Mycobacterium spp., very low in finished waters, occurred idiosyncratically at high levels in some tap waters, indicating distribution and/or premise plumbing as main contributors of mycobacteria. Legionella spp., another genus containing potential opportunistic pathogens, also occurred ubiquitously. Source water microbiology was most divergent from tap water, and each step of treatment brought samples more closely similar to tap waters.
Little research has been conducted on creative, eminent women across different domains, perceptions of their own creativity, and what drives their creative productivity. This research used ...qualitative, comparative case study methodology to investigate 15 creative women who were identified as highly innovative across diverse domains. Interviews were conducted and biographical information collected about each eminent woman's work in various fields including: Literature, History, Psychology, Biology, Genomics, Pharmacology, Engineering, Business and Art. The study investigated paths taken and choices made over decades as these women reached high levels of creative achievement and made transformational contributions in their field or domain. Each woman believed she was creative, had strong creative self-efficacy, and explained her drive to create as deriving from love and passion for her chosen work. These creative women actively strived and managed obstacles to serve as agents of positive change, transform or improve their disciplines, and use their creativity to make a positive impact and difference in the world. Their experiences provide insights into the decisions and strategies that enabled them to achieve high levels of innovation, which may help inform and inspire other women in the pursuit of creative productive giftedness.
Objective: Review the short- and long-term outcomes of a single institution experience in infants with congenital tracheal stenosis, comparing four different operative techniques used from 1982 ...through 2000. Methods: Hospital and clinic records of 50 infants and children who had surgical repair of congenital tracheal stenosis secondary to complete tracheal rings were reviewed. Age at surgery ranged from 7 days to 72 months (median, 5 months, mean 7.8±12 months). Techniques included pericardial patch tracheoplasty (n=28), tracheal autograft (n=12), tracheal resection (n=8), and slide tracheoplasty (n=2). All procedures were done through a median sternotomy with cardiopulmonary bypass. Seventeen patients had a pulmonary artery sling (35%), and 11 had an intracardiac anomaly (22%). Results: There were three early deaths (6% early mortality), two after pericardial tracheoplasty and one after autograft. There were six late deaths (12% late mortality), five after pericardial tracheoplasty and one after slide tracheoplasty. Length of stay (median) was 60 days (pericardial tracheoplasty), 28 days (autograft), 14 days (resection), and 18 days (slide). Reoperation and/or stent placement was required in seven patients (25%) after pericardial tracheoplasty, in two patients (17%) after autograft, in no patients after resection, and in one patient (50%) after slide tracheoplasty. Conclusions: Our current procedures of choice for infants with congenital tracheal stenosis are resection with end-to-end anastomosis for short-segment stenoses (up to eight rings) and the autograft technique for long-segment stenoses. Associated pulmonary artery sling and intracardiac anomalies should be repaired simultaneously.
Objective Prediction of surgical site infection and mortality after cardiac surgery might allow for interventions to reduce adverse outcomes. We sought to evaluate existing risk scores. Methods We ...included 809 consecutive patients undergoing coronary artery bypass surgery. Data were collected prospectively. Infections were defined as deep sternal wound infection or mediastinitis by using established criteria and evaluated 60 days after surgical intervention. All-cause mortality was assessed at 30 days and 6 months. We assessed the ability of the National Nosocomial Infections Surveillance risk index, the EuroSCORE, and the Society of Thoracic Surgeons risk score to predict infection and mortality. Discrimination was assessed using the area under the receiver operating curve. Results The rate of surgical site infection was 3.6% (29/809 patients). The National Nosocomial Infections Surveillance risk index showed moderate discrimination for infection (area under the receiver operating curve of 0.64) and poor ability to stratify patients into infection risk groups. The EuroSCORE predicted infection and 30-day and 6-month mortality with good discrimination (area under the receiver operating curve of 0.72, 0.78, and 0.77, respectively). Ranking patients by the EuroSCORE and dividing the cohort into 3 roughly equal risk groups yielded an ascending risk for infection of 0.7%, 3.0%, and 7.2%. The preoperative and intraoperative Society of Thoracic Surgeons risk scores showed good discrimination for surgical site infection (area under the receiver operating curve of 0.72 and 0.76, respectively) and excellent discrimination for early and late mortality (area under the receiver operating curve of >0.80). Risk grouping based on the Society of Thoracic Surgeons score yielded an ascending risk for infection of 0.7%, 3.6%, and 6.4%. Conclusions The EuroSCORE and the Society of Thoracic Surgeons risk score can be used for joint risk stratification for surgical site infection and mortality. Both scores performed better than the National Nosocomial Infections Surveillance risk index.
•Mixing corn pellets into compost of rooting areas tends to increase its overall use.•Mixing corn pellets into compost of rooting areas increases its use as lying area.•Time of day and temperature ...influence pig behaviour in a rooting area.•Mixing corn pellets into compost of rooting areas does not affect the areas’ cleanliness.
Exploratory behaviour is an essential part of the behavioural repertoire of pigs. Providing pigs with a rooting area filled with appropriate material enables such behaviour and is therefore considered to improve animal welfare. Managing the hygiene of a rooting area, however, is often challenging when pigs use it also for elimination. Mixing corn pellets into the rooting material could increase use and exploration while reducing elimination behaviour. To investigate this hypothesis, we constructed rooting areas filled with compost produced from garden waste in four pens on a commercial organic farm. We compared two experimental pens (E) with rooting areas filled with compost, in which we mixed 2 kg of corn pellets every morning, with two control pens (C, rooting areas filled with compost only). The experiment started in October 2019 and lasted 34 weeks with seven replicates in total. Group size ranged between 21–35 pigs (N = 386). We registered behaviour once a week through direct observations of the complete outdoor area and additional video recordings of the rooting area. Behavioural variables of interest were activity status (i.e. standing/sitting or lying), rooting, agonistic and play behaviour. We assessed cleanliness of the rooting material via visual scoring and chemical analysis of compost samples. The latter included tests on dry matter content, conductivity, and ammonium concentration. Data were analysed with linear mixed-effects models. Results showed that there was a tendency for a higher total number of pigs in the rooting area in E than in C (p = 0.06). In E, more pigs were lying in the rooting area than in C (p = 0.04). There was no difference between treatments in rooting behaviour. In addition, the overall use of the outdoor run did not differ between treatments. Time of day influenced all recorded behaviours in the rooting area (p < 0.001). With increasing temperature, more pigs were present in the outdoor run (p < 0.001) and in the rooting area (p < 0.01) for both treatments. Conductivity and ammonium concentration in the compost increased the longer the compost remained in the rooting area (p < 0.001), but there was no difference between the two treatments. We conclude that mixing corn pellets into rooting material increases the use of the rooting area by heightening the overall presence of pigs in it but not its cleanliness.
Williams Syndrome (WS) is a neurodevelopment disorder associated with a hemizygous deletion on chromosome 7. WS is characterized with mental retardation, severe visual–spatial deficits, relative ...language preservation, and excellent facial recognition. Distinctive auditory features include musical ability, heightened sound sensitivity, and specific patterns of auditory evoked potentials. These features have led to the hypothesis that the dorsal forebrain is more affected than the ventral. Previously, we reported primary visual area 17 abnormalities in rostral striate cortex, a region contributing to the dorsal visual pathway. Based on the dorsal–ventral hypothesis, and language and auditory findings, we predicted a more normal histometric picture in auditory area 41. We used an optical dissector method to measure neurons in layers II–VI of area 41 in right and left hemispheres of the same 3 WS and 3 control brains used in the area 17 study. There was a hemisphere by diagnosis interaction in cell packing density (CPD) in layer IV and in cell size in layer III between WS and control brains. Post hoc analysis disclosed in control brains, but not WS, a layer IV left > right asymmetry in CPD, and a layer III left < right asymmetry in cell size. WS brains showed more large neurons bilaterally in layer II and in left layer VI. Histometric alterations in area 41 were less widespread than rostral visual cortex. Also, there was less asymmetry in the WS brain. We interpret layers II and VI differences as reflecting increased limbic connectivity in primary auditory cortex of WS.
Abstract Objective The aim of the study is to review the safety and efficacy of partial fistulectomy with 3-layered primary closure without postoperative intubation for persistent tracheocutaneous ...fistula (TCF). Design This is a retrospective study. Setting The study was conducted in a tertiary care pediatric hospital. Patients Records of 49 children treated for persistent TCF between 1996 and 2005 were reviewed. Patients were studied if the TCF was closed using a 3-layered primary closure, and they had at least 1 year of follow-up. Thirty-nine patients met inclusion criteria. Results All patients were extubated immediately after surgery. Drains were removed, and all patients were discharged on the first postoperative day. The most common indications for tracheostomy were prolonged intubation and subglottic stenosis. There were 2 major and 3 minor complications. One major complication involved subcutaneous emphysema that developed on the seventh postoperative day because of cough. The other involved a poorly controlled diabetic patient who developed a postoperative infection with dehiscence. All fistulas remained closed at follow-up. Conclusion Partial excision and primary closure of persistent TCF is safe and effective. Drain placement and overnight observation are imperative. Careful patient selection is important. Routine postoperative intubation is not necessary.
Identification of protein-protein interactions is essential for elucidating the biochemical mechanism of signal transduction. Purification and identification of individual proteins in mammalian cells ...have been difficult, however, due to the sheer complexity of protein mixtures obtained from cellular extracts. Recently, a tandem affinity purification (TAP) method has been developed as a tool that allows rapid purification of native protein complexes expressed at their natural level in engineered yeast cells. To adapt this method to mammalian cells, we have created a TAP tag retroviral expression vector to allow stable expression of the TAP-tagged protein at close to physiological levels. To demonstrate the utility of this vector, we have fused a TAP tag, consisting of a protein A tag, a cleavage site for the tobacco etch virus (TEV) protease, and the FLAG epitope, to the N terminus of human SMAD3 and SMAD4. We have stably expressed these proteins in mammalian cells at desirable levels by retroviral gene transfer and purified native SMAD3 protein complexes from cell lysates. The combination of two different affinity tags greatly reduced the number of nonspecific proteins in the mixture. We have identified HSP70 as a specific interacting protein of SMAD3. We demonstrated that SMAD3, but not SMAD1, binds HSP70 in vivo, validating the TAP purification approach. This method is applicable to virtually any protein and provides an efficient way to purify unknown proteins to homogeneity from the complex mixtures found in mammalian cell lysates in preparation for identification by mass spectrometry.
Objectives: Evaluate the results of a technique for repair of congenital tracheal stenosis by use of a free tracheal autograft.
Methods: Between January 1996 and July 1997, six infants with ...congenital tracheal stenosis resulting from complete tracheal rings underwent repair with a free tracheal autograft. Mean age at the time of repair was 4.9 months; mean weight was 5.4 kg. The approach was through a median sternotomy with cardiopulmonary bypass for respiratory support. The trachea was incised anteriorly through the area of stenosis, the midportion of the stenotic trachea was excised, and an end-to-end anastomosis was carried out posteriorly. The excised tracheal segment (1.3 to 2.2 cm long) was used as a free autograft to patch the lower trachea anteriorly. In four infants the autograft was augmented in the upper trachea with pericardium; in two patients with a shorter length of stenosis, the autograft completed the repair. Simultaneous pulmonary artery sling repair (4), ligation and division of patent ductus arteriosus (3), cricoid split (2), atrial and ventricular septal defect repair (1), and complete atrioventricular canal repair (1) were performed at the time of tracheal repair.
Results: The infants were extubated and discharged at a mean of 13 and 23 days postoperatively, respectively. One infant had recurrent tracheal stenosis related to the pericardial patch and required a tracheal stent and tracheostomy 4 months postoperatively. Our mean follow-up is 11 months. Bronchoscopic findings currently show widely patent tracheal lumina in all infants.
Conclusions: The technique of free tracheal autograft with and without pericardial augmentation was successful in opening the airway of six infants with congenital tracheal stenosis and is currently our procedure of choice for children with this diagnosis. (J Thorac Cardiovasc Surg 1998;115:869-74)
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