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Interactions between fused, a segment-polarity gene in Drosophila, and other segmentation genes.

Fused (fu) is a segment polarity gene whose product is maternally required in the posterior part of each segment. To define further the role of fused and determine how it interacts with other segmentation genes, we examined the phenotypes obtained by combining fused with mutations of pair rule, homeotic and other segment polarity loci. When it was possible, we also looked at the distribution of corresponding proteins in fused mutant embryos. We observed that fused-naked (fu;nkd) double mutant embryos display a phenotypic suppression of simple mutant phenotypes: both naked cuticle and denticle belts, which would normally have been deleted by one of the two mutants alone, were restored. In fused mutant embryos, engrailed (en) and wingless (wg) expression was normal until germ band extension, but partially and completely disappeared respectively during germ band retraction. In the fu;nkd double mutant embryo, en was expressed as in nkd mutant at germ band extension, but later this expression was restricted and became normal at germ band retraction. On the contrary, wg expression disappeared as in fu simple mutant embryos. We conclude that the requirements for fused, naked and wingless activities for normal segmental patterning are not absolute, and propose mechanisms by which these genes interact to specify anterior and posterior cell fates.

Animals↗

Microsatellite analysis provides evidence of neoplastic transformation in long-segment, but not in short-segment, Barrett's oesophagus.

It has been suggested that the high prevalence of short segments of specialised intestinal metaplasia (SIM) at the gastro-oesophageal junction is associated with the rising incidence of oesophageal adenocarcinoma. Our aims were to document the prevalence of short segments of SIM at the gastro-oesophageal junction in patients attending for routine endoscopy and to determine if there was molecular evidence of neoplastic transformation in those with SIM. Patients (n = 101) were recruited from randomly selected upper gastro-intestinal endoscopy lists. Biopsy specimens were taken at the squamo-columnar junction to assess the prevalence of SIM. Frozen sections were assessed for molecular evidence of neoplastic transformation using microsatellite analysis. Squamo-columnar biopsies were suitable for analysis in 95 patients, of whom 20 (21%) had oesophagitis and 2 (2%) had Barrett's oesophagus (>3 cm of endoscopically apparent columnar-lined oesophagus). Twenty patients had SIM at the gastro-oesophageal junction, including 2 with Barrett's oesophagus and 18 with short segments of SIM, one of whom had an associated intramucosal adenocarcinoma detected incidentally by histology. Three of the 20 cases with SIM exhibited novel microsatellite alleles, 2 with Barrett's oesophagus and 1 with short segment SIM and an associated adenocarcinoma. The 18 patients with short segments of SIM at the gastro-oesophageal junction were significantly older than those without SIM.

Adenocarcinoma↗

Chemical synthesis of human beta-endorphin(1-27) analogs by peptide segment coupling. Leucine and glycine residues bearing thiocarboxyl functions as junctions for peptide segment coupling.

[Gly8]beta hEP(1-27)NH2 and [L-Leu8]beta hEP(1-27)NH2, two analogs of human beta-endorphin, were synthesized by both all-stepwise solid phase synthesis and peptide segment coupling. For the peptide segment coupling method, two thiocarboxyl peptides. Msc-[Gly8]beta hEP(1-8)SH and Msc-[L-Leu8]beta hEP(1-8)SH, were synthesized by standard solid phase method on 4-[alpha-(Boc-Gly-S)benzyl]phenoxyacetamidomethy-resin and 4-[alpha-(Boc-L-Leu-S)benzyl]phenoxyacetamidomethy-resin. These two thiocarboxyl peptides were coupled to H-[Lys(Cit)9,19,24]-beta hEP(9-27)NH2. [Gly8]beta hEP(1-27)NH2 and [L-Leu8]beta hEP(1-27)NH2 were obtained after removal of Msc groups and citraconyl groups from products of the segment coupling reaction. The yields of both [Gly8]beta hEP(1-27)NH2 and [L-Leu8]beta hEP(1-27)NH2 in the segment coupling reaction were approximately 18%. Less than 1% of racemization of Leu-8 occurred during coupling of Msc-[L-Leu8]beta hEP(1-8)SH to H-[Lys(Cit)9,19,24]-beta hEP(9-27)NH2. Results of amino acid composition analysis, analysis by reverse phase high pressure liquid chromatography and receptor binding activity assays of the analogs showed that peptide analogs prepared by segment coupling method and those prepared by all-stepwise solid phase synthesis were identical. Results of receptor binding activity assays suggested that the molecular charge properties of beta-endorphin(1-27) and its analogs influenced the receptor binding activity.

Amino Acid Sequence↗

Interposition of 'wide' cycloperistaltic segments into isolated segments of jejunum--influence on fluid perfusion.

Two isolated jejunal segments (Thiry-Vella (T-V) loops) were created in 7 dogs. One contained a 'wide' cycloperistaltic (C-P) segment while the other, acting as a control, did not. The C-P segment had the same diameter as the jejunum. Fluid perfusion experiments showed that there were no changes in the volume of output, absorption and pooling of the fluid in the T-V loops containing the 'wide' C-P segment. This was in contrast with significant changes shown in earlier experiments using 'narrow' C-P segments.

Animals↗

Focal-segmental glomerulosclerosis. The relationship between tubular atrophy and segmental sclerosis.

Biopsy specimens with focal-segmental glomerulosclerosis (FSGS) show segmental sclerosis (SS) and tubular atrophy (TA). The relationship between SS and TA was studied using serial sections. Based on the location of the SS, 3 and 5 biopsy specimens, respectively, were classified as FSGS, glomerular tip lesion (GTL) or FSGS, not otherwise specified (NOS). The proximal tubule was classified as normal or atrophic. The glomerulus was tracked through serial sections in both directions and classified as normal, SS (graded 1(+)-4+), or ischemic. Segmental sclerosis was identified in 21 of 24 glomeruli in FSGS, GTL and in 46 of 89 glomeruli in FSGS, NOS; TA was identified in 13 of 16 glomeruli in FSGS, GTL and 6 of 13 glomeruli in FSGS, NOS, which showed 1+ SS, and in 36 of 38 glomeruli in both forms, which showed 2+ to 3+ SS. Basement membrane disruptions at the glomerular-tubular junction and interstitial expansion with inflammation was encountered. Three biopsy specimens with FSGS, NOS had hypertensive changes and showed TA in several normal and in all ischemic glomeruli. Chronic tubular injury is present in most glomeruli containing segmental lesions, even small lesions, suggesting that the glomerular and tubular injury may have a common cause or that tubular injury may result from constituent(s) in the glomerular filtrate.

Adult↗

Effect of soft segment chemistry on the biostability of segmented polyurethanes. I. In vitro oxidation.

A series of segmented polyurethanes (SPUs) containing various polyol soft segments was prepared and their resistance to oxidative degradation was investigated after aging in AgNO3 solution. The SPU with the polyether soft segment showed a large reduction in mechanical strength after exposure to the oxidative environment. Surface cracking was often observed for these specimens. XPS measurements revealed that scission of the ether linkage occurs upon oxidation. The oxidative resistance of SPUs containing aliphatic hydrocarbon soft segments was significantly improved over the poly(tetramethylene oxide) (PTMO) based polyurethane.

Calorimetry, Differential Scanning↗

Analyses of segment-specific expression of alkaline phosphatase activity in the mesoderm of the oligochaete annelid Tubifex: implications for specification of segmental identity.

In the embryos of the oligochaete annelid Tubifex, segments VII and VIII specifically express mesodermal alkaline phophatase (ALP) activity in the ventrolateral region. In this study, we examined whether this segment-specific expression of ALP activity depends on external cues. Cell lineage analyses show that the ALP-expressing cells originate from M teloblasts. Furthermore, a set of teloblast-ablation experiments demonstrated that the seventh and eighth primary m blast cells (m7 and m8) produced from M teloblasts give rise to ALP-expressing cells in segments VII and VIII, respectively, and that primary m blast cells other than m7 and m8 lack the ability to generate ALP-expressing progeny cells. The results of another set of blastomere-ablation experiments suggest that ALP-expressing cells emerge independently of interactions with surrounding tissues. Teloblast-transplantation experiments demonstrated that m8 can generate ALP-expressing cells in an ectopical position, suggesting that it is unlikely that ALP activity emerges in response to the positional cues residing in the embryo. These results suggest that m7 and m8 are exclusively specified as precursors of ALP-expressing cells at the time of their birth from M teloblasts. We propose that segmental identities in primary m blast cells of the Tubifex embryo are determined according to the genealogical position in the M lineage and that the M teloblast possesses a developmental program through which the sequence of blast cell identities is determined.

Alkaline Phosphatase↗

Cadherin-defined segments and parasagittal cell ribbons in the developing chicken cerebellum.

In the developing chicken cerebellar cortex, three cadherins (Cad6B, Cad7, and R-cadherin) are expressed in distinct parasagittal segments that are separated from each other by ribbons of migrating interneurons and granule cells which express R-cadherin and Cad7, respectively. The segment/ribbon pattern is respected by the expression of other types of molecules, such as engrailed-2 and SC1/BEN/DM-GRASP. The cadherin-defined segments contain young Purkinje cells which are connected to underlying nuclear zones expressing the same cadherin, thereby forming parasagittal cortico-nuclear zones of topographically organized connections. In addition, R-cadherin-positive mossy fiber terminals display a periodic pattern in the internal granular layer. In this layer, Cad7 and R-cadherin are associated with synaptic complexes. These results suggest that cadherins play a pivotal role in the formation of functional cerebellar architecture by providing a three-dimensional scaffold of adhesive information.

Animals↗

Ultrastructure of the kidney of a South American caecilian, Typhlonectes compressicaudus (Amphibia, Gymnophiona). I. Renal corpuscle, neck segment, proximal tubule and intermediate segment.

The ultrastructure of the renal corpuscle, the neck segment, the proximal tubule and the intermediate segment of the kidney of a South American caecilian, Typhlonectes compressicaudus (Amphibia, Gymnophiona) was examined by means of transmission electron microscopy (TEM), scanning electron microscopy (SEM) and freeze-fracture technique. The glomerular filter apparatus consists of the podocyte epithelium, a distinct basement membrane, a subendothelial space and the capillary endothelium. Emanating from the podocyte cell body, several long primary processes encircle neighboring capillaries. The short slender foot processes originating from the primary processes interdigitate with those from other primary processes, thereby forming the meandering filtration slit. Thick bundles of microfilaments are found in the primary processes, but absent in the foot processes. The basement membrane consists of a lamina rara externa and a rather thin lamina densa (50 nm thickness). The wide subendothelial space contains abundant microfibrils, a few collagen fibrils and many thin processes of mesangial cells. The endothelium is flat and fenestrated (compared to mammals displaying relatively few fenestrations); some of the fenestrations are bridged by a diaphragm. The glomerular mesangium is made up of the mesangial cells and a prominent mesangial matrix containing microfibrils and collagen fibrils. The cells of the neck and intermediate segments display numerous cilia with their microtubules arranged in the typical 9 + 2 pattern. The basal bodies of the cilia are attached to thick filaments with a clear crossbanding pattern of 65 nm periodicity. The proximal tubule is composed of cells typical for this segment (PT cells) and light cells lacking a brush border (bald-headed cells). The PT cells measure 10-25 micron in height and 15-30 micron in width and do not interdigitate at their lateral borders with each other. Their basolateral cell membrane is amplified by many folds projecting into lateral intercellular spaces and into basal recesses. The brush border is scarce and composed of loosely arranged short microvilli.

Amphibians↗

Spurious ST segment depression by automated ST segment analysis.

Perioperative myocardial ischemia has been shown to be predictive of adverse cardiac outcome [1]. One method of detecting intraoperative myocardial ischemia is automated ST segment monitoring [2]. The rationale behind automated ST monitoring is that the clinician will be promptly alerted to ischemic changes in the ECG. This may lead to earlier intervention and, perhaps, improved outcome [3]. We describe a case in which automated ST segment monitoring falsely signaled the presence of intraoperative ST segment depression. This event emphasizes the importance of confirming the validity of ST segment changes before instituting therapy.

Aged↗

Evaluation of normal variations in S-T segment patterns by body surface isopotential mapping: S-T segment elevation in absence of heart disease.

S-T segment elevation is commonly observed in the electrocardiogram of normal persons. To study the possible origins of such patterns, 45 normal volunteers were examined. Electrocardiographic potentials were registered from 150 torso electrodes and processed to construct isopotential maps at 2 ms intervals throughout the QRS-T interval. The maximal potentials recorded from any of the 150 electrodes were 198 +/- 76.4 and 272.1 +/- 84.2 microV at instants 40 and 80 ms into the S-T segment, respectively. Maximal voltages recorded by the six standard precordial V leads at these respective time points were 109.7 +/- 57.0 and 163.6 +/- 66.9 microV. Torso maximal potentials were significantly stronger than were those sensed by V leads; the two were significantly correlated but predictability was limited. The duration of overlap between the onset of ventricular recovery and the end of the excitation was determined from isopotential maps and ranged rom 4 to 16 ms. There was no significant correlation (p greater than 0.05) between these values and either torso or V lead potentials at either 40 or 80 ms into S-T segment. These data suggest that (1) standard precordial leads do not accurately predict maximal torso potentials during the S-T segment, and (2) the degree of overlap between repolarization and depolarization is not a major determinant of precordial voltage. Hence, the rationale for use of the term "early repolarization" to describe this clinical condition is not substantiated.

Adolescent↗

Visual conspicuity of a moving dot, horizontal line segment or vertical line segment.

The effect of varying the retinal eccentricity of a moving target on its conspicuity were explored for targets whose shape, orientation, size and luminance were the same as the stationary background elements. The target was a dot, horizontal line segment, or vertical line segment. The main results were: (1) conspicuity increased monotically as a function of velocity for all of the cases. (2) The rate of the increase of conspicuity slowed down at around a velocity between 0.6 and 0.8 deg/sec (break in slope). (3) For individual subjects, the size of conspicuity area for a dot was not so different from that for a horizontal line segment, but it was very different from that for a vertical line segment. (4) The position of the break in slope shifted upward or downward from one another but not laterally. The results indicate that the conspicuity was not due to the sum of the outputs in movingness analyzing system (MAS, Transient-channel) and displacement analyzing system (DAS, Sustained-channel), but due to one of them according to velocity of the moving element.

Fixation, Ocular↗

Putative full-length clones of the genomic DNA segments of subterranean clover stunt virus and identification of the segment coding for the viral coat protein.

Subterranean clover stunt disease is an economically important aphid-borne virus disease affecting certain pasture and grain legumes in Australia. The virus associated with the disease, subterranean clover stunt virus (SCSV), was previously found to be representative of a new type of single-stranded DNA virus. Analysis of the virion DNA and restriction mapping of double-stranded cDNA synthesized from virion DNA suggested that SCSV has a segmented genome composed of 3 or 4 different species of circular ssDNA each of about 850-880 nucleotides. To further investigate the complexity of the SCSV genome, we have isolated the replicative form DNA from infected pea and from it prepared putative full-length clones representing the SCSV genome segments. Analysis of these clones by restriction mapping indicated that clones representing at least 4 distinct genomic segments were obtained. This method is thus suitable for generating an extensive genomic library of novel ssDNA viruses containing multiple genome segments such as SCSV and banana bunchy top virus. The N-terminal amino acid sequence and amino acid composition of the coat protein of SCSV were determined. Comparison of the amino acid sequence with partial DNA sequence data, and the distinctly different restriction maps obtained for the full-length clones suggested that only one of these clones contained the coat protein gene. The results confirmed that SCSV has a functionally divided genome composed of several distinct ssDNA circles each of about 1 kb.

Amino Acid Sequence↗

Computer model analysis of the relationship of ST-segment and ST-segment/heart rate slope response to the constituents of the ischemic injury source.

The objective of the study was to investigate a proposed linear relationship between the extent of myocardial ischemic injury and the ST-segment/heart rate (ST/HR) slope by computer simulation of the injury sources arising in exercise electrocardiographic (ECG) tests. The extent and location of the ischemic injury were simulated for both single- and multivessel coronary artery disease by use of an accurate source-volume conductor model which assumes a linear relationship between heart rate and extent of ischemia. The results indicated that in some cases the ST/HR slope in leads II, aVF, and especially V5 may be related to the extent of ischemia. However, the simulations demonstrated that neither the ST-segment deviation nor the ST/HR slope was directly proportional to either the area of the ischemic boundary or the number of vessels occluded. Furthermore, in multivessel coronary artery disease, the temporal and spatial diversity of the generated multiple injury sources distorted the presumed linearity between ST-segment deviation and heart rate. It was concluded that the ST/HR slope and ST-segment deviation of the 12-lead ECG are not able to indicate extent of ischemic injury or number of vessels occluded.

Adult↗

Lamina VII and VIII neurons of the S2 segment bilaterally projecting to the C6 segment of the spinal cord in the cat.

Intracellular and extracellular recordings of antidromic action potentials were applied to investigate neurons of the S2 segment projecting to the C6 segment of the cat spinal cord. The cell bodies were located in laminae VII and VIII of the gray matter while axons ascended in lateral funiculi. Thirty-two out of the total 45 neurons were found to project to the C6 segment bilaterally, seven ipsilaterally and six contralaterally. The axonal conduction velocities were in the 42-96 m/s range and in some neurons were significantly lower in distal parts of axons, supposing that some neurons may give off collateral branches to various segments of the spinal cord. It is discussed if the investigated neurons form a part of the propriospinal system or if their cervical projections are only collaterals of long tracts ascending to supraspinal levels. The organisation of the presented connections between spinal enlargements indicates their contribution in complex mechanisms of co-ordination of movements of the limbs.

Action Potentials↗

NMR observation of selected segments in a larger protein: central-segment isotope labeling through intein-mediated ligation.

Peptide segments in a protein, which can include an active site of interest or be a series of parts constituting the entire structure, are now selectively observed by nuclear magnetic resonance (NMR) spectroscopy using samples prepared by the intein-mediated ligation method. Two separate inteins were used to ligate NMR-transparent segments to both the ends of an NMR-visible segment, producing a partly visible intact protein molecule. The (15)N-(1)H correlation spectrum of a 370-residue maltose binding protein labeled with (15)N at a continuous segment comprising residues Gly(101)-Ser(238) showed the essential elimination of signal overlapping, the signals being at the same positions as for the uniformly labeled sample. This method will allow structural analysis by NMR of over 50-kDa proteins in combination with contemporary NMR techniques suppressing the signal decays of larger proteins.

ATP-Binding Cassette Transporters↗

ST-segment depression in lead aVR predicts predischarge left ventricular dysfunction in patients with reperfused anterior acute myocardial infarction with anterolateral ST-segment elevation.

BACKGROUND: Patients with an anterolateral acute myocardial infarction (AMI) have a worse prognosis, and those with additional inferolateral wall involvement might be higher risk because of more extensive area at risk. Lead -aVR obtained by inversion of images in lead aVR has been reported to provide useful information for inferolateral lesion. METHODS: We examined the relation between ST-segment deviation in lead aVR on admission electrocardiogram (ECG) and left ventricular function in 105 patients with an anterolateral AMI undergoing successful reperfusion < or = 6 hours after onset. Patients were classified according to ST-segment deviation in lead aVR on admission ECG: group A, 23 patients with ST elevation of > or = 0.5 mm; group B, 47 patients without ST deviation; and group C, 35 patients with ST depression of > or = 0.5 mm. RESULTS: There were no differences among the 3 groups in age, sex, or site of the culprit lesion. In groups A, B, and C, the peak creatine kinase level was 3661 +/- 1428, 4440 +/- 1889, and 6959 +/- 2712 mU/mL, and the left ventricular ejection fraction (LVEF) measured by predischarge left ventriculography was 54% +/- 9%, 48% +/- 7%, and 37% +/- 9%, respectively(P < .01). During hospitalization, congestive heart failure occurred more frequently in group C than in groups A or B (P < .05). ST-segment depression in lead aVR had a higher predictive accuracy than other ECG findings in identifying patients with predischarge LVEF < or = 35%. CONCLUSIONS: We conclude that in patients with an anterolateral AMI, ST-segment depression in lead aVR on admission ECG is useful for predicting larger infarct and left ventricular dysfunction despite successful reperfusion.

Adult↗

Multiple DNA and protein sequence alignment based on segment-to-segment comparison.

In this paper, a new way to think about, and to construct, pairwise as well as multiple alignments of DNA and protein sequences is proposed. Rather than forcing alignments to either align single residues or to introduce gaps by defining an alignment as a path running right from the source up to the sink in the associated dot-matrix diagram, we propose to consider alignments as consistent equivalence relations defined on the set of all positions occurring in all sequences under consideration. We also propose constructing alignments from whole segments exhibiting highly significant overall similarity rather than by aligning individual residues. Consequently, we present an alignment algorithm that (i) is based on segment-to-segment comparison instead of the commonly used residue-to-residue comparison and which (ii) avoids the well-known difficulties concerning the choice of appropriate gap penalties: gaps are not treated explicity, but remain as those parts of the sequences that do not belong to any of the aligned segments. Finally, we discuss the application of our algorithm to two test examples and compare it with commonly used alignment methods. As a first example, we aligned a set of 11 DNA sequences coding for functional helix-loop-helix proteins. Though the sequences show only low overall similarity, our program correctly aligned all of the 11 functional sites, which was a unique result among the methods tested. As a by-product, the reading frames of the sequences were identified. Next, we aligned a set of ribonuclease H proteins and compared our results with alignments produced by other programs as reported by McClure et al. [McClure, M. A., Vasi, T. K. & Fitch, W. M. (1994) Mol. Biol. Evol. 11, 571-592]. Our program was one of the best scoring programs. However, in contrast to other methods, our protein alignments are independent of user-defined parameters.

Algorithms↗