Dynamics and organization of MAP kinase signal pathways

In the budding yeast, Saccharomyces cerevisiae, four separate but structurally related mitogen-activated protein kinase (MAPK) activation pathways are known. The best understood of these regulates mating. Pheromone binding to receptor informs cells of the proximity of a mating partner and induces differentiation to a mating competent state. The MAPK activation cascade mediating this signal is made up of Ste11 (a MEK kinase [MEKK]), Ste7 (a MAPK/ERK kinase [MEK]), and the redundant MAPK-related Fus3 and Kss1 enzymes. Another MAPK activation pathway is important for cell integrity and regulates cell wall construction. This cascade consists of Bck1 (a MEKK), the redundant Mkk1 and Mkk2 enzymes (MEKs), and Mpk1 (a MAPK). We exploited these two pathways to learn about the coordination and signal transmission fidelity of MAPK activation cascades. Two lines of evidence suggest that the activities of the mating and cell integrity pathways are coordinated during mating differentiation. First, cells deficient in Mpk1 are susceptible to lysis when they make a mating projection in response to pheromone. Second, Mpk1 activation during pheromone induction coincides with projection formation. The mechanism underlying this coordination is still unknown to us. Our working model is that projection formation generates a mobile second messenger for activation of the cell integrity pathway. Analysis of a STE7 mutation gave us some unanticipated but important insights into parameters important for fidelity of signal transmission. The Ste7 variant has a serine to proline substitution at position 368. Ste7-P368 has higher basal activity than the wild-type enzyme but still requires Ste11 for its function. Additionally, the proline substitution enables the variant to transmit the signal from mammalian Raf expressed in yeast. This novel activity suggests that Ste7-P368 is inherently more permissive than Ste7 in its interactions with MEKKs. Yet, Ste7-P368 cross function in the cell integrity pathway occurs only when it is highly overproduced or when Ste5 is missing. This behavior suggests that Ste5, which has been proposed to be a tether for the kinases in the mating pathway, contributes to Ste7 specificity and fidelity of signal transmission.     

Reaction products of ammonium nitrate phosphate fertilizers of varying water-soluble phosphorus content in different Indian soils

Studies were undertaken on the isolation and identification of reaction products of ammonium nitrate phosphate (ANP) fertilizers containing 30, 50 and 70 per cent water-soluble phosphorus (WSP) of total phosphorus in representative soils of the vertisol, oxisol, alfisol, entisol, mollisol and aridisol groups of India. ANP fertilizers were applied in solid form to soil, and reaction products formed at and around the site of ANP fertilizer placement were identified after six weeks incubation in moist soils by X-ray diffraction technique. DCPD (dicalcium phosphate dihydrate- CaHPO₄ · 2H₂O) was the major reaction product of ANP fertilizers containing 30 and 50 per cent WSP in vertisol, entisol, aridisol, mollisol, oxisol and alfisol, and of ANP containing 70 per cent WSP in vertisol, entisol, alfisol, aridisol and mollisol. DCP (dicalcium phosphate-CaHPO₄) was detected with ANP of 30 and 50 per cent WSP in the vertisol, alfisol, entisol, mollisol and aridisol groups of soils. In addition to DCPD, FePO₄ · 2H₂O (metastrengite) and AlPO₄ · 2H₂ O-monoclinic (metavariscite) were formed in alfisol and oxisol soils with ANP of 30 and 50 per cent WSP. FePO₄ · 2H₂O and AlPO₄ · 2H₂O (metavariscite) were identified in alfisol and oxisol soils while AlPO₄ · 2H₂O-orthorhombic (variscite) was formed in alfisol soils with ANP of 70 per cent WSP.     

Interpretation time of serial chest CT examinations with stacked-metaphor workstation versus film alternator

PURPOSE: Interpretation time of serial staging chest CT cases, which each contained current and previous examinations, with a simple prototype workstation called filmstack was experimentally compared with interpretation time with a film alternator. MATERIALS AND METHODS: The filmstack displayed a "stack" of sections for each examination; user controls allowed rapid selection of preset attenuation windows and both synchronized and unsynchronized scrolling. Eight radiologists were timed as they used the filmstack and the film alternator to interpret four ergonomically complex serial CT cases. RESULTS: All reports dictated on the basis of findings with filmstack and film were of acceptable clinical accuracy. The time to examine a case with filmstack was significantly faster than the time with film, including the time to load and unload the alternator (99% confidence [P = .01]). There was no statistically significant difference in interpretation time between filmstack and prehung film. CONCLUSION: Use of a low-cost stacked CT workstation with a single 1,024 x 1,024 monitor is an effective means of interpreting cases that require comparison of multiple CT examinations.     

Augmentation of facial soft-tissue defects with Alloderm dermal graft

The aim of the study was TPS diagnostic value determination in the serum of women with breast cancer as compared with MCA and CA 15-3. The relationship between the serum concentration of these antigens and patient age, clinical stage, histological grade, presence of metastases to lymph nodes and histological type of neoplasm was evaluated. Studies were conducted on the sera of 139 women before surgical procedure aged 28-81, treated in the Clinic of Oncology, at Karol Marcinkowski University of Medical Sciences in Poznań. The TPS concentration was determined using the "BEKI Diagnostics" immunoenzymatic method, MCA - by the "Roche" test and CA 15-3 concentration was determined by the "Abbott" immunofluorescent test. The study showed significantly higher levels of TPS, CA 15-3 and MCA in women with cancer, compared with values in healthy women and women with mastopathy. The highest median of concentration and frequency of occurrence was obtained for TPS. A correlation between enhancement of TPS and CA 15-3 concentration with clinical stage was observed. A similar connection was noted in women with metastases to the lymph nodes. Serum MCA concentration results did not demonstrate the above effects. The study suggests, that in estimating the clinical condition of women with breast cancer, the simultaneous determination of TPS with CA 15-3 seems to be a more useful prognostic factor than TPS or CA 15-3 with MCA.     

Legal framework of postgraduate nursing education in spain

Being part of the first report of the SEEIUC Forum on the training of nurses in critical care units, this article shows the different postgraduation training paths which Spanish legislation establishes. The "Titulos Oficiales de Especialización Profesional" ("Official Degrees on Professional Specialization") settle the seven nursing specialties regulated by the Decreto 992/1987. Following a second path, "Titulos de Postgraduado no Oficiales" ("Non-official postgraduation degrees"), every University acknowledged by the LRU and creating them as their Own Degrees, may organize Master courses, University experts, University specialists and Postgraduation university degrees, according to their autonomy. So that this autonomous offer is as homogeneous as possible, there is an interuniversity agreement which encompasses 24 national universities and gathers the general criteria for the academic organization of such courses. The report is completed by an analysis of the training offer for critical care nursing, developed during the 1995/1996 course in Spain.     

The lactulose breath hydrogen test and small intestinal bacterial overgrowth

OBJECTIVES: To i) document the sensitivity and specificity of a combined scintigraphic/lactulose breath hydrogen test for small intestinal bacterial overgrowth and ii) investigate the validity of currently accepted definitions of an abnormal lactulose breath hydrogen test based on "double peaks" in breath hydrogen concentrations. METHODS: Twenty-eight subjects were investigated with culture of proximal small intestinal aspirate and a 10-g lactulose breath hydrogen test combined with scintigraphy. Gastroduodenal pH, the presence or absence of gastric bacterial overgrowth, and the in vitro capability of overgrowth flora to ferment lactulose were determined. RESULTS: Sensitivity (16.7%) and specificity (70.0%) of the lactulose breath hydrogen test alone for small intestinal bacterial overgrowth were poor. Combination with scintigraphy resulted in 100% specificity, because double peaks in serial breath hydrogen concentrations may occur as a result of lactulose fermentation by cecal bacteria. Sensitivity increased to 38.9% with scintigraphy, because a single rise in breath hydrogen concentrations, commencing before the test meal reaches the cecum, may occur in this disorder. Sensitivity remained suboptimal irrespective of the definition of small intestinal bacterial overgrowth used, the nature of the overgrowth flora, favorable luminal pH, the presence of concurrent gastric bacterial overgrowth, or the in vitro ability of the overgrowth flora to ferment lactulose. CONCLUSIONS: Definitions of an abnormal lactulose breath hydrogen test based on the occurrence of double peaks in breath hydrogen concentrations are inappropriate. Not even the addition of scintigraphy renders this test a clinically useful alternative to culture of aspirate for diagnosing small intestinal bacterial overgrowth.     

The morphological mechanism of the development of myosatellitocytes from the structural elements of the muscle fiber during increased functional activity of the skeletal muscles

Under increased muscular activity in some muscular fibers disintegration areas of myofibrillar apparatus has been revealed. Migration of myonuclei into these microregions starts the mechanism of their segregation due to plasmolemma produced from the reticulum sarcoplasmaticum and triad systems surface. After plasmolemma production in "sarcocytes" intensive development and differentiation of organellae occur. As a result of differentiation "sarcocytes" transform in to myosatellitocytes of type-2 and migrate under lamina externa muscular fibers. So, a hypothesis about formation of myogenic tissue's cellular phase from the myosymplastic one has been confirmed.     

Definition of inseparably fused ventricular myocardium in thoracopagus: fetal echocardiographic utility and pathologic refinement

Correlative echocardiographic and pathological findings in a thoracopagus with conjoined hearts are reported. One twin had tricuspid atresia with discordant atrioventricular connections and concordant ventriculoarterial connections. The morphologic right ventricle was hypoplastic and there was a large muscular ventricular septal defect. The other twin had hypoplasia of the mitral valve anulus and left ventricle with double-outlet right ventricle and pulmonary valve atresia. The tricuspid valve was severely insufficient in part because of a large orifice and redundant, elongated leaflets with abnormal chordal attachments. The left ventricles of these two twins shared a perforated common "free wall" with at least two large defects allowing mixing of the circulations at that level. Not all anatomic details were established conclusively by fetal echocardiography; however, sufficient diagnostic information was obtained to support a decision not to aggressively resuscitate these twins after elective cesarean delivery at 31 weeks' gestation.     

Selective isolation of reversible cold sensitive variants from Chinese hamster ovary cell cultures

The fine structure and cellular associations of the large pigment cells (LPC's) of the compound eye of the house fly were studied with high voltage and conventional electron microscopy. Depending on the sector of the compound eye, the facets are either rectangular or hexagonal. The underside of each facet has indentations exactly aligned with those on top into which inserts an angulated sleeve of LPC's. Under the rectangular lens facet 6 or 8 small compact (in cross section) LPC's join four elongate LPC's. Clusters of compact cells alternate in this ring with elongate ones. Compact cells compress together and become quadrangular (in cross section) several microns below their insertion into the lens and form "building block" corners while elongate cells form "side rails" for the rectangular type of distal pseudocone enclosure. Beneath hexagonal facets all LPC's are rather elongate with out corner cells. In both facet types LPC's enclose the pseudocone for a longitudinal distance of 4 mum and then are displaced as bordering cells by a sleeve of two corneal pigment cells (CPC's), each of which encloses half of the proximal pseudocone. For the following 6 mum of longitudinal distance these concentric sleeves of CPC's and LPC's form a double layer around the pseudocone. At about 10 mum below lens base the two sleeves separate; LPC's become attenuated and extend cable-like to the basement membrane and CPC's enclose the proximal pseudocone, Semper cells and distal retinula. The junction between lens and LPC's has critical structural value in that (1) this is the sole anchorage to the lens by the lengthy remainder of the ommatidium, and (2) LPC's enclose the semiliquid pseudocone in the most distal portion of the pseudocone. In addition to vertical support, the LPC's send out numerous lateral processes that make structural contact among themselves, with the corneal pigment cells and the photoreceptor cells. The structural features of this array are discussed relative to possible physiological roles.     

A cytosolic activity distinct from crm1 mediates nuclear export of protein kinase inhibitor in permeabilized cells

The leucine-rich nuclear export signal (NES) is used by a variety of proteins to facilitate their delivery from the nucleus to the cytoplasm. One of the best-studied examples, protein kinase inhibitor (PKI), binds to the catalytic subunit of protein kinase A in the nucleus and mediates its rapid export to the cytoplasm. We developed a permeabilized cell assay that reconstitutes nuclear export mediated by PKI, and we used it to characterize the cytosolic factors required for this process. The two-step assay involves an import phase and an export phase, and quantitation is achieved by digital fluorescence microscopy. During the import phase, a fluorescent derivative of streptavidin is imported into the nuclei of digitonin-permeabilized HeLa cells. During the export phase, biotinylated PKI diffuses into the nucleus, binds to fluorescent streptavidin, and mediates export of the complex to the cytoplasm. Nuclear export of the PKI complex is cytosol dependent and can be stimulated by addition of the purified NES receptor, Crm1. HeLa cell cytosol treated with N-ethylmaleimide (NEM) or phenyl-Sepharose to inactivate or deplete Crm1, respectively, is still fully active in the PKI export assay. Significantly, the export activity can be depleted from cytosol by preadsorption with a protein conjugate that contains a functional NES. These data indicate that cytosol contains an export activity that is distinct from Crm1 and is likely to correspond to an NES receptor.