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PubMed1996

Identification of the major membrane and core proteins of vaccinia virus by two-dimensional electrophoresis.

Vaccinia virus assembly has been well studied at the ultrastructural level, but little is known about the molecular events that occur during that process. Towards this goal, we have identified the major membrane and core proteins of the intracellular mature virus (IMV). Pure IMV preparations were subjected to Nonidet P-40 (NP-40) and dithiothreitol (DTT) treatment to separate the core proteins from the membrane proteins. These proteins were subsequently separated by two-dimensional (2D) gel electrophoresis, and the major polypeptide spots, as detected by silver staining and 35S labeling, were identified by either matrix-assisted laser desorption/ionization mass spectrometry, N-terminal amino acid sequencing, or immunoprecipitation with defined antibodies. Sixteen major spots that partitioned into the NP-40-DTT-soluble fraction were identified; 11 of these were previously described virally encoded proteins and 5 were cellular proteins, mostly of mitochondrial origin. The core fraction revealed four major spots of previously described core proteins, two of which were also detected in the membrane fraction. Subsequently, the NP-40-DTT-soluble and -insoluble fractions from purified virus preparations, separated by 2D gels, were compared with postnuclear supernatants of infected cells that had been metabolically labeled at late times (6 to 8 h) postinfection. This relatively short labeling period as well as the apparent shutoff of host protein synthesis allowed the selective detection in such postnuclear supernatants of virus-encoded proteins. These postnuclear supernatants were subsequently treated with Triton X-114 or with sodium carbonate to distinguish the membrane proteins from the soluble proteins. We have identified the major late membrane and nonmembrane proteins of the IMV as they occur in the virus as well as in infected cells. This 2D gel map should provide an important reference for future molecular studies of vaccinia virus morphogenesis.

Amino Acid Sequence↗

PubMedDate not supplied

Feasibility of intensity-modulated radiation therapy in the treatment of advanced cervical chordoma.

AIMS AND BACKGROUND: Postoperative radiation is often given in cases of cervical chordoma because of the high incidence of local recurrence. The tumor mass usually surrounds the spinal cord and infiltrates vertebral bone. A combined technique using protons or electrons to boost the initial photon fields is generally applied. We evaluated the use of dynamic intensity-modulated radiation therapy as an alternative technique for treating advanced cervical chordoma. METHODS AND STUDY DESIGN: A female patient with incomplete resection of a vertebral chordoma surrounding C2-C3 was irradiated with a total dose of 58 Gy (ICRU point) in 2 Gy daily fractions for 29 days between December 2001 and January 2002. Beam arrangement consisted of seven 6 MV non-opposed coplanar fields. Pretreatment quality assurance included checking of the absolute dose at reference points and 2D dose map analysis. Treatment was delivered with a 120-leaf collimator in sliding window mode. To verify the daily setup, portal images at 0 degrees and 90 degrees were compared with the simulation images before treatment delivery (manual matching) and after treatment delivery (automatic anatomy matching). RESULTS AND CONCLUSIONS: The mean dose to the planning target volume (PTV) was 57.6 +/- 2.1 Gy covering 95% of the PTV per 95% isodose. The minimum dose to the PTV (D99) was 53.6 Gy in the overlapping area between the PTV and the spinal cord planning organ at risk volume (PRV). The maximum dose to the spinal cord was 42.2 Gy and to the spinal cord PRV (8 mm margin) 53.7 Gy. The mean dose to the parotid glands was 37.4 Gy (homolateral gland) and 19.5 Gy (contralateral gland). Average deviation in setup was -1.1 +/- 2.5 mm (anterior-posterior), 2.4 +/- 1.3 mm (latero-lateral), 0.7 +/- 0.9 mm (craniocaudal) and -0.43 +/- 1 degree (rotation). CONCLUSIONS: In the treatment of chordomas surrounding the spinal cord, intensity-modulated radiotherapy can provide high dose homogeneity and PTV coverage. Frequent digital portal image-based setup control is able to reduce random positioning errors for head and neck cancer patients immobilized with conventional thermoplastic masks.

Chordoma↗

PubMed2006

Analysis of the prostate cancer cell line LNCaP transcriptome using a sequencing-by-synthesis approach.

BACKGROUND: High throughput sequencing-by-synthesis is an emerging technology that allows the rapid production of millions of bases of data. Although the sequence reads are short, they can readily be used for re-sequencing. By re-sequencing the mRNA products of a cell, one may rapidly discover polymorphisms and splice variants particular to that cell. RESULTS: We present the utility of massively parallel sequencing by synthesis for profiling the transcriptome of a human prostate cancer cell-line, LNCaP, that has been treated with the synthetic androgen, R1881. Through the generation of approximately 20 megabases (MB) of EST data, we detect transcription from over 10,000 gene loci, 25 previously undescribed alternative splicing events involving known exons, and over 1,500 high quality single nucleotide discrepancies with the reference human sequence. Further, we map nearly 10,000 ESTs to positions on the genome where no transcription is currently predicted to occur. We also characterize various obstacles with using sequencing by synthesis for transcriptome analysis and propose solutions to these problems. CONCLUSION: The use of high-throughput sequencing-by-synthesis methods for transcript profiling allows the specific and sensitive detection of many of a cell's transcripts, and also allows the discovery of high quality base discrepancies, and alternative splice variants. Thus, this technology may provide an effective means of understanding various disease states, discovering novel targets for disease treatment, and discovery of novel transcripts.

Adenocarcinoma↗

PubMed1982

Detection and visualization of regurgitant flow in valvular diseases by pulsed Doppler technique.

Regurgitant flows in valvular diseases were evaluated by a pulsed Doppler flowmeter combined with an electronic beam sector scanning echocardiograph. The apparatus which was newly developed by us allowed the simultaneous demonstration of a sample site on a two-dimensional echocardiogram with flow measurement. Doppler signals of regurgitant flow were recorded as uni-directional or bi-directional wide frequency band signals. The locations, where regurgitant flow signals were detected, were depicted on the corresponding two-dimensional echocardiogram. This procedure was referred to as "a flow mapping technique" for non-invasive visualization of the distribution of regurgitant flow. In 12 patients with mitral regurgitation due to mitral valve prolapse detected by the pulsed Doppler technique, the regurgitant flow was distributed to the opposite side of the prolapsing mitral leaflet. The transmission of the regurgitant murmur was well consistent with the direction of the regurgitant flow. In 14 patients with aortic regurgitation, the distribution of aortic regurgitant flow visualized by the flow mapping technique closely coincided with that obtained by cineaortography. Based on the distribution of the regurgitation, the severity of the regurgitation could be precisely evaluated by the Doppler technique. Regurgitant flow signals were detected in the right atrium in all 13 patients with tricuspid regurgitation diagnosed by right ventriculography. We found 4 patients who did not show Carvallo's sign but in whom were detected regurgitant flow signals by the Doppler technique. In all of them, tricuspid regurgitation was proven at surgery. These results indicate that the Doppler technique presented here has an obvious clinical advantage in detecting and evaluating regurgitant flow in valvular diseases.

Adult↗

PubMed2005

[Loxosceles Heinecken & Lowe, 1835 (Araneae; Sicariidae) species distribution in the State of Paraná].

The State of Paraná registers on average 2,577 loxoscelic accidents annually. For the elaboration of control and management programs one should first determine the distribution of the species of the genus Loxosceles. A mapping was performed of Loxosceles references in various scientific collections. A total of 1,561 spiders were found, identified as Loxosceles intermedia (67%), Loxosceles gaucho (19.5%), Loxosceles laeta (10.8%) and Loxosceles hirsuta (2.4%), originating from 20 regional and 69 municipal health districts. Loxosceles intermedia was present in all areas of the state (50 municipal districts), while Loxosceles gaucho occurred in the north and northwest, (17 municipal districts), Loxosceles laeta in the south (13 municipal districts) and Loxosceles hirsuta in the west and central areas (10 municipal districts). Paraná has four of the eight species of Loxosceles registered in Brazil. Given the medical importance of accidents caused by these spiders, it is necessary to perform studies on the location of such incidents and investigate areas that have not yet been sampled.

Animals↗

PubMedDate not supplied

Nursing phenomena identified in family planning visits with ICNP - Beta version 2.

This descriptive, exploratory, retrospective survey, carried out at a family planning service, aimed to identify nursing phenomena during nursing visits according to the ICNP, Beta version 2. Data were collected based on 52 records of nursing visits, realized from October 2001 to December 2002. To conduct the cross-mapping process, all identified nursing phenomena were joined, organized and compared according to the ICNP's terms. Of the 51 identified nursing phenomena/diagnoses, 46 (90.2%) showed exact and partial concordance. The identified nursing phenomena can be used to assist nurses to provide care for clients in family planning services. The ICNP showed to be a comprehensive program, although some terms need to be reviewed and others enhanced. However, considering that it is an international classification applicable to several countries, the mapping process and cross-references were very satisfactory.

Family Planning Services↗

PubMed1997

Navigational aids for real-time virtual bronchoscopy.

OBJECTIVE: Virtual bronchoscopy is a recently described technique for performing simulated bronchoscopy using data obtained from thin-section CT scans of the airways. Although the detail visible during virtual bronchoscopy can be striking, the lack of global context and physical cues can lead to disorientation for the radiologist, especially during real-time "fly-throughs." We investigated the use of seven software tools (coordinate axes, global maps, trail markers, cross-references to standard two-dimensional projections, rear-view windows, collision avoidance, and stereoscopic displays) that are navigational aids to overcome such disorientation. CONCLUSION: The navigational aids performed satisfactorily on virtual bronchoscopy studies from 21 patients. A frame rate of one to seven frames per second, depending on the complexity of the bronchial model, could be attained. Several of the aids worked best when used selectively or when updated frequently but not continuously. Navigational aids are potentially useful for guiding radiologists who do virtual bronchoscopy studies. Such aids are general tools that may be applicable to other types of virtual endoscopy.

Adult↗

PubMed1993

Business process analysis. Part 2.

We are addressing in this three part series, the challenge of making lasting, logical and linked improvements in your organization, aided by fresh, comprehensive and almost certainly unique, new view(s) of your organizational system. Preparing your organization for this type of needed "reinvention" of the organizational system can be better assured if some, preferably all, of the conditions listed in Figure 1 are present. These were introduced and discussed in last month's article. The last item in Figure 1 refers to a "Business Process Map" of the organization. Chances are that you have never encountered this term and only have a vague idea of what is meant by it. Explaining the basis of this evaluative approach is the purpose for this article. Next month, we will show ho useful this insight can be in many strategic decision areas of your business.

Decision Making, Organizational↗

PubMed1999

Projections from striate and extrastriate visual cortices of the cat to the reticular thalamic nucleus.

We have studied the pattern of connectivity of the visual cortical areas 17, 18, 19, 20a, 21a, posteromedial lateral (PMLS), and the posterolateral lateral (PLLS) suprasylvian areas with the reticular thalamic nucleus (RTN) of the cat ventral thalamus. Three cortical areas per hemisphere were injected iontophoretically with either 4% wheat germ agglutinin-horseradish peroxidase, 4% dextran-fluororuby, or 4% dextran-biotin. The visual field representations of the injection sites were determined by reference to previously published visuotopic maps of the cortex. The locations of labelled fibres, presumed terminals and cell bodies were determined with the aid of a camera lucida attachment and computer aided stereometry. In the ventral thalamus, the primary visual cortices (areas 17 and 18) project in a topographic manner to both the perigeniculate nucleus (PGN) and the RTN. By contrast, the "higher" visual cortical areas (areas 19, 21a, 20a, PMLS, and PLLS) project only to the RTN. Our experiments demonstrate the existence of a single, albeit coarse, visuotopic map within the RTN but do not support the notion of separate subregions within the RTN that can be related specifically to a particular visual cortical area. The putative single visuotopic map in the RTN appears to be organised in such a way that the vertical meridians are represented along the rostrocaudal axis of the RTN, whereas the horizontal meridians are mapped within the dorsoventral axis of the nucleus. The upper visual field is represented within regions of the RTN adjacent to the caudal part of the dorsal lateral geniculate nucleus (LGNd), whereas the lower visual field is represented in the parts of the RTN rostral to the LGNd. The map also shows a ventrodorsal shift along the rostrocaudal axis of the RTN such that in the rostral RTN the representation of vertical meridian is placed more ventrally than that in the caudal part of the nucleus.

Animals↗

PubMed1999

Loss of PTEN expression in paraffin-embedded primary prostate cancer correlates with high Gleason score and advanced stage.

The tumor suppressor gene PTEN/MMAC-1/TEP-1 (referred to hereafter as PTEN) maps to chromosome 10q23 and encodes a dual specificity phosphatase. The PTEN protein negatively regulates cell migration and cell survival and induces a G1 cell cycle block via negative regulation of the phosphatidylinositol 3'-kinase/protein kinase B/Akt signaling pathway. PTEN is frequently mutated or deleted in both prostate cancer cell lines and primary prostate cancers. A murine polyclonal antiserum was raised against a glutathione S-transferase fusion polypeptide of the COOH terninus of PTEN. Archival paraffin tissue sections from 109 cases of resected prostate cancer were immunostained with the antiserum, using DU145 and PC-3 cells as positive and negative controls, respectively. PTEN expression was seen in the secretory cells. Cases were considered positive when granular cytoplasmic staining was seen in all tumor cells, mixed when areas of both positive and negative tumor cell clones were seen, and negative when adjacent benign prostate tissue but not tumor tissue showed positive staining. Seventeen cases (15.6%) of prostate cancer were positive, 70 cases (64.2%) were mixed, and 22 cases (20.2%) were negative. Total absence of PTEN expression correlated with the Gleason score (P = 0.0081) and correlated more significantly with a Gleason score of 7 or higher (P = 0.0004) and with advanced pathological stage (American Joint Committee on Cancer stages T3b and T4; P = 0.0078). Thus, loss of PTEN protein is correlated with pathological markers of poor prognosis in prostate cancer.

Adult↗

PubMedDate not supplied

Renal effects of antenatally or postnatally administered steroids.

Steroids administrated antenatally to the mothers improve postnatal outcomes of the newborns with pleiotropic effects. Furthermore steroids have been used in preterm infants to prevent or treat chronic lung disease. Synthetical glucocorticoids readily cross placental barrier and reach significant pharmacologic levels in the fetus: besides their well known pulmonary effects they have a concomitant maturational effect of postnatal renal function in preterm infants both with a direct and indirect effect. Endogenous and exogenous glucocorticoids play a role in the maintenance of glomerular filtration (GFR). The antenatal administration of steroids increases the GFR, in association to the maturation of the tubular function. According to different studies the improvement of renal function, expressed by the increase of GFR, is only partially referable to the increase of MAP and the improvement of the cardiovascular status, while it was imputable to a direct renal effect of the steroids, especially on the renal blood flow, on functional glomerular surface area available for filtration and on the glomerular filtrate of the single cortical nephron. However debate remains about the mechanism through which steroids would act on the renal vascular smooth muscolature. The increase the GFR observed after the antenatal administration of glucocorticoids in premature fetuses is also accompanied by an increase of urinary flow and of fractional excretion of sodium. Glucocorticoids would increase the proximal reabsorption of sodium increasing directly the function and the expression of the sodium transporters and both indirectly and directly increasing the activity of Na-K-ATPase. In extremely low weight antenatal administration of betamethasone or dexamethasone was associated with lower estimated insensible water loss, secondary to a direct maturational effect in the skin epithelial barrier, as well as an increased reabsorption of the fetal lung fluid. Moreover antenatal glucocorticoid administration was associated, at birth, to a significant suppression of plasma renin activity and angiotensin II in comparison to the controls. Despite the wide use of the steroidal therapy in the prevention of the bronchopulmonary dysplasia, only few articles, in literature, analyse the effects of glucocorticoids on postnatal renal function, such as the increase in urinary flow. The authors think that steroids contribute in a meaningful way to the clinical improvement observed in children with BPD through the maturative action on the premature kidney with effect both at glomerular and tubular level.

Female↗

PubMed2004

Comparison of echocardiographic methods in assessing severity of mitral regurgitation in patients with mitral valve prolapse.

BACKGROUND AND AIM OF THE STUDY: Mitral regurgitation (MR) shows different characteristics in mitral valve prolapse (MVP); hence, it is important to assess MR severity accurately in these patients. The study aim was to compare Doppler echocardiographic methods in making such assessment. METHODS: Forty-seven patients with confirmed MVP and at least moderate mitral insufficiency, as established by Doppler echocardiography, were studied. Quantitative Doppler was used as the reference standard method. Color Doppler mapping was used to determine regurgitant jet area (JA/LAA), flow convergence (EROA-PISA) and vena contracta width (VCW). Systolic pulmonary venous flow reversal (SPVFR) and mitral E-wave velocity were also monitored. RESULTS: Univariate analysis showed severe MR to be significantly correlated to age, presence of atrial fibrillation, left ventricular systolic and diastolic diameter, left atrial diameter, mitral E velocity, JA/LAA, VCW, EROA-PISA and the presence of SPVFR. On multivariate analysis, the strongest determinants of severe MR were EROA-PISA, VCW and E velocity. The greatest area under the receiver-operator curve for diagnosing severe MR was observed with EROA-PISA. The 45-mm2 threshold of EROA-PISA had the highest risk ratio of severe MR with a high sum of sensitivity and specificity. However, the JA/LAA had the lowest risk ratio and negative predictive value for severe MR. CONCLUSION: PISA, VCW, E velocity and SPVFR measurements may be used to evaluate MR severity semi-quantitatively in patients with MVP; however, the ratio of JA/LAA appears to be a less reliable method in this respect.

Adolescent↗

PubMed1983

Distribution of abnormal Q waves on body surface in relation to left ventricular wall motion abnormalities in myocardial infarction.

This study was undertaken to determine the possible one-to-one relationship between each site of asynergy of the left ventricle and the body surface area to which ensuing abnormal electrical phenomena are reflected. In 140 post-myocardial infarction (MI) patients, distribution of abnormal Q waves on the body surface was correlated with the abnormal segments of LV wall motion identified by left ventriculography (LVG). Unipolar lead electrocardiograms (ECGs) were recorded from 87 lead points over the precordium and the back with Wilson's central terminal as the reference point. Data acquisition and mapping was accomplished through a mapping system HPM 5100 microcomputer. In all cases, coronary arteriography (CAG) and LVG were performed at least 2 months after the acute episode of MI. The LVG findings were evaluated separately in seven wall segments in accordance with the American Heart Association (AHA) reporting system. Sensitivity, specificity, diagnostic accuracy, positive predictive value, and negative predictive value of the abnormal Q wave in each lead point were obtained in cases with abnormal wall motion in segments 2, 3, and 6, respectively, and in infero-posterior segments 4 or 5. Statistical analysis was performed by comparing two groups of patients with or without asynergy of each wall segment concerned. Results of the study revealed some abnormal Q areas of high diagnostic accuracy that correlated highly with the site of abnormal contractility.

Journal Article↗

PubMed1991

Morphometry of the neonatal fetal alcohol syndrome face from 'snapshots'.

Recognition of alcohol-related birth defects is less successful in the neonate than in older children. Clarren et al. (1987) were able to describe the geometry of the FAS face at age 7. To define neonatal facial features of FAS, 3 reference points were used to map 7 frontal and 7 profile landmarks coded as standardized cartesian coordinates. These landmarks were digitized from snapshots of 21 neonates, prospectively diagnosed as FAS blinded for prenatal alcohol exposure, and 76 non-FAS controls. In the controls, race was found to be a significant determinant of landmarks, as expected. Using discriminant analysis and controlling for race, the FAS face was characterized by landmark measurements which correspond to short palpebral fissure, scooping out of the nasal bridge and thin vermilion [F(4,71)-7.8, r sq-30%, p < .01]. Jackknifed sensitivity was 75%, specificity, 79%. These findings suggest that FAS-defining features can be quantified in the neonate via computer-based screening of infants from at-risk pregnancies.

Alcoholism↗

PubMed1982

A SERLINE-based union list of serials for basic health sciences libraries: a detailed protocol.

In March 1981 the Consortium for Information Resources (CIR) was chosen by the Massachusetts Health Sciences Library Network to develop and automate a statewide biomedical union list of serials. Employing a commercial processor, ANSI standard Z39.42-1980, and SERLINE, CIR consolidated the journal holdings of six Massachusetts health-related library consortia. SERLINE, with its unique identifier as the single control element, governed the form of entry and bibliographic data for each journal. Additionally, SERLINE enhanced the union list by providing "see references" and general notations to map users to main titles or special information. An original feature of this union list is the "rolled" holdings and location statements intended to encourage even distribution of interlibrary loan transactions. The resulting union list of serials includes the holdings of 116 Massachusetts libraries, 94 of which are hospital libraries. The list includes nearly 3,000 unique titles and 15,000 holdings statements; production costs averaged $1.35 per unique title and 27 per holdings statement.

Information Systems↗

PubMed1993

[Structure-activity relationship of synthetic and semisynthetic opioid agonists and antagonists].

This account presents the chemical structure-pharmacological activity relationships of semi-synthetic morphine and synthetic morphinan and benzomorphan derivatives. Substitution of nitrogen confers the analgesic or morphine antagonist activity. Opiate receptor selectivity of these compounds is also discussed. The importance of morphine agonists, and antagonists is mentioned with particular reference to the opiate receptor mapping.

Molecular Structure↗

PubMed1996

[Biochemical approaches to neuromuscular junction disorders].

In view of the conformation-dependent B-cell epitope and the MHC class II-restricted T-cell epitope, myasthenogenic sites in the molecular structure of acetylcholine receptor alpha-subunit were defined by the induction of a rat model of myasthenia gravis using synthetic peptides, alpha 183-200, alpha 125-147 and alpha 67-76 with alpha 107-116. Structural model for the acetylcholine receptor synthetic peptides were designed to obtain a conformation that was highly antigenic compared with that of natural sequence peptides. To search for antigenic sites in the molecular structure of P/Q type of voltage-gated calcium channel (alpha 1) in Lambert-Eaton myasthenic syndrome (LEMS), LEMS patients' sera were studied by immunoprecipitation assay using antigens synthesized referring to its sequential residue map. Peptides corresponding to extracellular region (S5 S6 linker) of domains II and IV were specifically reactive with LEMS antibodies; their titers respectively correlated with those of anti-P/Q type calcium channel (human cerebellum). The extracellular region of synaptotagmin, a synaptic vesicle protein that acts as a calcium sensor in release of acetylcholine, was also immunogenic in the induction of a rat model of LEMS; a proportion of LEMS patients' sera reacted with recombinant synaptotagmin in immunoblot.

Animals↗

PubMed1997

The alpha and omega of G-protein coupled receptors: a novel method for classification. Part 2. Bin classification.

A novel way of classification of G-protein coupled receptors is presented that is only based on receptor sequence information by counting of amino acid residues. It involves the number of amino acid residues between the Asn residue in TM1 and the residue Cys in the loop between TM4 and TM5, the number of residues between the latter Cys residue and Pro residue in TM6, and the number of residues between the latter Pro and the last amino acid residue (called omega) in the sequence. The classification of 131 sequences, covering biogenic amine, opioid and somatostatin receptors, is visualized by means of a diagram which is referred to as a bin map. Each bin in the diagram encloses all the sequences that belong to one and only one receptor type or subtype. This so-called bin classification was obtained by means of the genetic algorithm methodology, which offers new opportunities for classifying proteins.

Animals↗