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A genetic linkage map of markers for human chromosome 20.

A continuous genetic linkage map with five polymorphic DNA markers, including one that defines a locus containing a variable number of tandem repeats (VNTR), has been constructed from genotypic analysis of 59 large reference families. The map spans a genetic distance of 105 cM in males and 115 cM in females and provides initial anchor points for a high-resolution map of human chromosome 20.

Alleles↗

Meta-PseU: A meta-classifier for robust prediction of RNA pseudouridine modification sites from long sequences.

BACKGROUND AND OBJECTIVES: Pseudouridine (Ψ) represents one of the most abundant and conserved RNA modifications. Ψ provides an additional hydrogen-bond donor that enhances RNA structural stability and modulates translation. It participates in diverse biological processes, including RNA-protein interactions, splicing, translational control, and stress responses. Aberrant pseudouridylation is implicated in cancer, neurodegenerative disorders, and autoimmune diseases. Despite its biological importance, experimental identification of Ψ sites remains time-consuming and costly, limiting the feasibility of transcriptome-wide profiling. Computational approaches have therefore become essential complements to experimental techniques. However, state-of-the-art machine-learning and deep-learning predictors often suffer from limited generalizability due to small training datasets. To overcome these issues, we aim at constructing new long-sequence datasets and developing a novel Ψ site predictor. METHODS: New long-sequence datasets were constructed as benchmarks for RNA Ψ-site prediction. The Ψ modification sites in RMBase 3.0 were mapped to the reference genomes across three species of human, mouse, and yeast, and the RNA sequences with a length of 201 were generated by extending the upstream and downstream from the mapped, central sites. To eliminate sequence redundancy, the sequences were clustered using CD-HIT with a 70% sequence identity threshold. We developed Meta-PseU, a logistic regression-based meta-classifier that considered 118 machine learning and deep learning classifiers. The datasets and programs are freely accessible at https://github.com/kuratahiroyuki/MetaPseU. RESULTS: By optimizing model configuration, we proposed the Meta-PseU model stacking 32 machine learning and deep learning classifiers out of 118 classifiers. Meta-PseU substantially improved model generalizability, overcoming a key limitation of existing approaches. It greatly outperformed state-of-the-art predictors and achieved increasing accuracy with increasing sequence length. CONCLUSIONS: Long-sequence datasets were newly constructed as benchmarks for RNA Ψ-site prediction. Meta-PseU offers a new framework for robust Ψ-site identification by using long sequences.

Pseudouridine↗

Assessing possible exposures of ground troops to Agent Orange during the Vietnam War: the use of contemporary military records.

BACKGROUND: Potential exposure of ground troops in Vietnam to Agent Orange and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) remains controversial despite the passage of 30 years since the Vietnam War. Because of uncertainty over the serum dioxin levels in ground troops at the end of their service in Vietnam, attempts have been made to develop a methodology for characterizing exposure of ground troops in Vietnam to Agent Orange and other herbicides based upon historical reconstruction from military records. Historical information is often useful in evaluating and modeling exposure, but such information should be reasonably accurate, complete, and reliable. METHODS: This paper reviews the procedures and supporting historical information related to the spraying of herbicides in Vietnam. The historical information is classified into two categories: procedural information and operational information. Procedural information covered the process and procedures followed in spraying herbicides from US Air Force fixed wing aircraft (Operation RANCH HAND) in Vietnam, and included approval procedures for spray missions, the criteria required to conduct a mission, the control exercised by the Forward Air Controller and the Tactical Air Control Center and the characteristics of the equipment used to apply the herbicides. Operational information includes data from the RANCH HAND Daily Air Activities Reports, which included geographic locations of specific spray missions, the amount of herbicide sprayed by a specific mission, reports of battle damage to spray aircraft, reports of fighter aircraft support for aerial spray missions, and any comments, such as reasons for canceling a mission. RESULTS: Historical information demonstrates that herbicide spray missions were carefully planned and that spraying only occurred when friendly forces were not located in the target area. RANCH HAND spray missions were either not approved or cancelled if approved when there were friendly forces in the area designated for spraying. Stringent criteria had to be met before spray missions could be approved. The operational information shows that spray missions for both defoliation and crop destruction were conducted in an extremely hostile environment. Heavy 'fighter suppression' with antipersonnel ordnance was used to minimize the impact of hostile ground fire on RANCH HAND aircraft. Procedures were in place that prohibited movement of troops into sprayed areas immediately after a mission due to the possible presence of unexploded ordnance delivered by fighter aircraft supporting RANCH HAND missions. The optimal nature of the spray equipment and application procedures minimized the possibility of significant spray drift. Conclusions. Few friendly troops were sprayed by fixed wing aircraft during Operation RANCH HAND, which delivered 95% of all defoliants used in Vietnam. Similarly, few troops were sprayed during helicopter or surface-based spray operations, which constituted the remaining 5% of defoliants. Detailed policies and procedures for approval and execution of spray missions ensured that friendly forces were not located in the areas targeted for spraying. Fighter aircraft assigned to accompany each spray mission frequently suppressed much of the hostile fire with bombs and other ordnance. Confirmed clearance of the target area was necessary to avoid friendly casualties. Historical records establish that these policies and procedures were strictly followed. Exposure of troops whether from direct spraying or movement through areas recently sprayed was very unlikely. The wartime military records of troop positions and herbicide operations are valuable for some purposes, but have specific limitations in exposure reconstruction. The completeness and accuracy of the geographic data (maps used by RANCH HAND and military ground units) were dependent upon the inherent precision of the map, the accuracy with which it depicted surface features, and the completeness and accuracy of the information on which it is based. Navigation by the crew using visual orientation and reference to the map was the only means that aircrew on spray missions had for establishing their locations. A Forward Air Controller independent of Operation RANCH HAND was present at the location of each spray target immediately before and during spraying operations to verify the target location and ensure that friendly forces were clear of the target area. Anecdotal reports of direct spraying of troops in Vietnam likely reflect the RANCH HAND missions spraying insecticide for mosquito control at regular intervals from March 1967 through February 1972. Outlook. The distribution and levels of serum dioxin in RANCH HAND veterans and the US Army Chemical Corps Vietnam veterans (the unit responsible for helicopter and ground-based spray operations) are distinguishable from typical levels in the population decades after the Vietnam conflict. An exposure model similar to that proposed in the 2003 report of the Institute of Medicine's Committee on 'Characterizing Exposure of Veterans to Agent Orange and Other Herbicides Used in Vietnam' was tested in 1988 by the Centers for Disease Control and Prevention and found to be a poor predictor of absorbed dose of TCDD. Military records during the Vietnam War lack the precision to determine that troops were directly sprayed with herbicides during Operation RANCH HAND, especially given the procedures in place to ensure clearance of friendly forces from the target area and the lack of elevated serum levels of TCDD in ground troops judged to have operated in heavily sprayed areas.

2,4,5-Trichlorophenoxyacetic Acid↗

Genome cross-referencing and XREFdb: implications for the identification and analysis of genes mutated in human disease.

Comparative genomics approaches and multi-organismal biology are valuable tools for genetic analysis. Cross-species connections between genes mutated in human disease states and homologues in model organisms can be particularly powerful, as model-organism gene function data and experimental approaches can shed light on the molecular mechanisms defective in the disease. We describe a project that is systematically identifying novel expressed sequence tag (EST) sequences that are highly related to genes in model organisms and mapping them to positions on the mouse and human maps. This process effectively cross-references model organism genes with mapped mammalian phenotypes, facilitating the identification of genes mutated in human disease states via the positional candidate approach. A public database, XREFdb (http:@www.ncbi.nlm.nih.gov/XREFdb/), disseminates similarity search, mapping and mammalian phenotype information and increases the rate at which these cross-species connections are established.

Animals↗

Detailed physical map of human chromosomal region 11q12-13 shows high meiotic recombination rate around the MEN1 locus.

We have constructed a physical map of the region q12-13 on chromosome 11 by combining data generated from a panel of radiation-reduced somatic cell hybrids and pulsed-field gel electrophoresis (PFGE). Twenty different genetic markers have been sublocalized and ordered within this region and a total of 8.0 megabases has been mapped in detail using rare-cutting restriction endonucleases and PFGE. In two instances, the long-range restriction PFGE map spans the total distance between pairs of loci that have been previously mapped by genetic linkage in reference families. Comparison of this physical map with the available linkage map indicates a great variation in the recombination frequency over the region. The recombination rate is higher than expected, particularly for markers flanking the MEN1 region. Thus, for the closest pair of linked markers on the centromeric side, one centimorgan corresponds to approximately 300 kilobases, and for markers on the telomeric side, one centimorgan corresponds to approximately 350-600 kilobases.

Animals↗

Two-dimensional gel electrophoresis maps of the proteome and phosphoproteome of primitively cultured rat mesangial cells.

Mesangial cells (MC) play an important role in maintaining the structure and function of the glomerulus. The proliferation of MC is a prominent feature of many kinds of glomerular disease. The first reference 2-DE maps of rat mesangial cells (RMC), stained with silver staining or Pro-Q Diamond dye, have been established here to describe the proteome and phosphoproteome of RMC, respectively. A total of 157 selected protein spots, corresponding to 118 unique proteins, have been identified by MALDI-TOF-MS or LC-ESI-IT-MS/MS, in which 37 protein spots representing 28 unique proteins have also been stained with Pro-Q Diamond, indicating that they are in phosphorylated forms. All the identified proteins were bioinformatically annotated in detail according to their physiochemical characteristics, subcellular location, and function. Most of the separated or identified protein spots are distributed in the area of mass 10-70 kDa and pI 5.0-8.0. The identified proteins include mainly cytoplasmic and nuclear proteins and some mitochondrial, endoplasmic reticulum, and membrane proteins. These proteins are classified into different functional groups such as structure and mobility proteins (21.2%), metabolic enzymes (16.9%), protein folding and metabolism proteins (13.6%), signaling proteins (14.4%), heat-shock proteins (7.6%), and other functional proteins (12.7%). While structure and mobility proteins are mostly represented by protein spots with high abundance, signaling proteins are mostly represented by protein spots with relatively low abundance. Such a 2-DE database for RMC, especially with many signaling proteins and phosphoproteins characterized, will provide a valuable resource for comparative proteomics analysis of normal and pathologic conditions affecting MC function or pathologic progress.

Animals↗

Spatial representation of maps.

Two experiments tested the hypothesis that the cognitive representation of spatial relations on a map is primarily a visually keyed process. In Experiment 1, undergraduates studied a labeled map where features were presented either as words alone, as mimetic drawings of the referent, or as geometric symbols. Cued recall data showed that discrete features are best located when they are semantically congruent with the label referent, a fact providing considerable support for the image position. Recognition testing required a set of introspective judgements ordinally related to the interfeature metric on the map. The judgment X distance estimates were highly correlated for all groups, intimating that a high correspondence exists between image distances and the original stimulus metric. Experiment 2 had undergraduates view either a full-scale or a three-quarter reduction of a reference map. Subjects in the reduced version were able to recognize significantly more interfeature comparisons, suggesting a predictable relation between visual and imaginal acuity. The judgement X distance results were essentially the same as in Experiment 1, and the two groups failed to differ in terms of cued recall. Data from both experiments imply that spatial relations are cognitively represented in at least a partially isomorphic fashion, which seems to possess quasi-pictorial qualities.

Cognition↗

A core linkage map of the bumblebee Bombus terrestris.

The bumblebee Bombus terrestris is an economically important pollinator and an emerging model species in quantitative and population genetics. We generated genetic linkage maps for 3 independent mapping populations of B. terrestris. The linkage map with the highest resolution had 21 linkage groups, which adequately represents the haploid chromosome number of B. terrestris (n = 18). This map can be considered saturated, with an average marker distance of 10.3 cM and an estimated genome coverage of 81%. Using flow cytometry, we have estimated the genome size of this species to be 625 Mb. With an estimated total recombination genome length of 2760 cM, this results in a ratio of 226 kb/cM between the physical and genetic genome sizes. A recurring set of microsatellites and amplified fragment length polymorphism (AFLP) markers allowed the alignment of 14 linkage groups between the 3 maps. We propose to adopt this core map as a reference tool for future genetic and molecular work in B. terrestris.

Animals↗

Radiofrequency ablation of arrhythmias guided by non-fluoroscopic catheter location: a prospective randomized trial.

AIMS: To compare the utility of non-fluoroscopic mapping systems (Carto and Ensite NavX) with that of conventional mapping in patients referred for catheter ablation of a wide variety of arrhythmias. METHODS AND RESULTS: Patients referred for catheter ablation (excluding atrial fibrillation, atypical atrial flutter, ventricular tachycardia in structural heart disease, and complete AV nodal ablation) were randomized equally to a procedure guided by Carto, Ensite NavX, or conventional mapping. A total of 145 patients were recruited (82 men, aged 49+/-16, range 18-85). In 19 patients, no ablation was performed, and in the remaining, typical atrial flutter, atrioventricular nodal re-entrant tachycardia, and atrioventricular re-entrant tachycardias [including Wolff-Parkinson-White (WPW)] accounted for 93% of ablations. Overall procedure time, immediate and short-term success, complication rate, and freedom from symptoms at follow-up were identical for all groups. NavX led to the least X-ray exposure: Navx vs. conventional, median (range): 4 (0-50) vs. 13 (2-46) min (P<0.001); NavX vs. Carto, median (range): 4 (0-50) vs. 6 (1-55) min (P=0.008). Both Carto and NavX increased disposable costs by 50% when compared with conventional (P<0.001). For typical atrial flutter, Carto and NavX reduced screening times without increasing procedure cost. If ablation was not performed, NavX was twice as expensive as Carto or conventional. CONCLUSION: Ensite NavX and Carto procedures have similar effectiveness and safety to a conventional approach; however, they both reduce X-ray exposure, with NavX producing a significantly greater effect than Carto. Although this benefit is achieved at a greater financial cost, there may be long-term benefits to catheter laboratory staff.

Arrhythmias, Cardiac↗

An improved probability mapping approach to assess genome mosaicism.

BACKGROUND: Maximum likelihood and posterior probability mapping are useful visualization techniques that are used to ascertain the mosaic nature of prokaryotic genomes. However, posterior probabilities, especially when calculated for four-taxon cases, tend to overestimate the support for tree topologies. Furthermore, because of poor taxon sampling four-taxon analyses suffer from sensitivity to the long branch attraction artifact. Here we extend the probability mapping approach by improving taxon sampling of the analyzed datasets, and by using bootstrap support values, a more conservative tool to assess reliability. RESULTS: Quartets of orthologous proteins were complemented with homologs from selected reference genomes. The mapping of bootstrap support values from these extended datasets gives results similar to the original maximum likelihood and posterior probability mapping. The more conservative nature of the plotted support values allows to focus further analyses on those protein families that strongly disagree with the majority or plurality of genes present in the analyzed genomes. CONCLUSION: Posterior probability is a non-conservative measure for support, and posterior probability mapping only provides a quick estimation of phylogenetic information content of four genomes. This approach can be utilized as a pre-screen to select genes that might have been horizontally transferred. Better taxon sampling combined with subtree analyses prevents the inconsistencies associated with four-taxon analyses, but retains the power of visual representation. Nevertheless, a case-by-case inspection of individual multi-taxon phylogenies remains necessary to differentiate unrecognized paralogy and shared phylogenetic reconstruction artifacts from horizontal gene transfer events.

Bacterial Proteins↗

[14C]N-ethylmaleimide labeling of the plasma membrane [H+]-ATPase of Neurospora crassa.

Treatment of the purified plasma membrane [H+]-ATPase of Neurospora crassa with the sulfhydryl reagent N-ethylmaleimide (NEM) leads to a marked inhibition of ATPase activity. Several lines of evidence indicate that inhibition is caused by the reaction of NEM with a single cysteine residue on the Mr = 104,000 polypeptide. (1) Inhibition by NEM follows pseudo-first order kinetics. (2) MgADP protects against NEM inactivation and at the same time decreases the incorporation of [14C]NEM into the Mr = 104,000 polypeptide. When "protectable" [14C]NEM incorporation is plotted as a function of inhibition of ATPase activity, a linear relationship is observed with a slope of 0.8. (3) Labeling of the ATPase with [14C]NEM can be restricted to the protectable site by pretreatment with cold NEM in the presence of MgADP, followed by an incubation with [14C]NEM in the absence of nucleotides. When this is done, and the enzyme is subjected to tryptic mapping and autoradiography, a single radioactive peptide fragment is found. The protectable site may yield information about the role of cysteine in the ATPase mechanism, and should also serve as a useful point of reference in enzyme mapping studies.

Adenosine Triphosphatases↗

A multiplex assay for the detection and mapping of complex glycerol kinase deficiency.

BACKGROUND: Glycerol kinase deficiency (GKD) is an X-linked recessive disorder that presents in both isolated and complex forms. The contiguous deletion that leads to GKD also commonly affects NR0B1 (DAX1), the gene associated with adrenal hypoplasia congenita, and DMD, the Duchenne muscular dystrophy gene. Molecular testing to delineate this deletion is expensive and has only limited availability. METHODS: We designed a multiplex PCR assay for the detection and mapping of a contiguous deletion potentially affecting the IL1RAPL1, NR0B1, GK, and DMD genes in a 29-month-old male patient with GKD. RESULTS: Multiplex PCR detected a contiguous deletion that involved the IL1RAPL1, NR0B1, GK, and DMD genes. Although the patient had a creatine kinase concentration within the reference interval, further mapping with PCR revealed that exon 74 was the last intact exon at the 3' end of the DMD gene. CONCLUSIONS: Multiplex PCR is an effective and inexpensive way to detect and map the contiguous deletion in cases of complex GKD. The extension of a deletion to include DMD exon 75 in a patient with a creatine kinase concentration within the reference interval suggests that this region of the gene may not be essential for protein function.

Child, Preschool↗

Patient specific treatment verifications for helical tomotherapy treatment plans.

We performed two-dimensional treatment verifications for ten patients planned and treated with helical tomotherapy. The treatment verification consisted of a film measurement as well as point dose measurements made with an ion chamber. The agreement between the calculated and the measured film dose distributions was evaluated with the gamma index calculated for three sets of criteria (2 mm and 2%, 4 mm and 3%, and 3 mm and 5%) as recommended in the literature. Good agreement was found between measured and calculated distributions without any need of normalization of the dose data but with dose map registration using reference marks. In this case, 69.8 +/- 17.2%, 92.6 +/- 9.0%, and 93.4 +/- 8.5% passed the 2 mm and 2%, 4 mm and 3%, and 3 mm and 5% criteria, respectively. Agreement was excellent when both normalization and manual registration of the dose maps was employed. In this case 91.2 +/- 5.6%, 99.0 +/- 1.4%, and 99.5 +/- 0.8% passed the 2 mm and 2%, 4 mm and 3%, and 3 mm and 5% criteria, respectively. The mean percent discrepancy for the point dose measurements was -0.5 +/- 1.1%, -2.4 +/- 3.7%, -1.1 +/- 7.3% for the high dose, low dose, and critical structure point, respectively. Three criteria for a satisfactory treatment verification in the high dose regions of a plan were established. For the un-normalized reference mark registered data 80% of pixels must pass the 3 mm and 5% criteria. For the normalized and manually registered data, 80% must pass the 2 mm and 2% criteria, and the point dose measurement must be within 2% of the calculated dose. All low dose region/critical structure point dose measurements were evaluated on a patient by patient basis. The criteria we recommend can be useful for the routine evaluation of treatment plans for tomotherapy systems.

Biophysical Phenomena↗

Physical and genetic mapping of the CDR gene with particular reference to its position with respect to the FRAXA site.

This study narrows down the localization of the gene coding for the cerebellar degeneration-related protein (CDR 34) to the upper boundary of the FRAXA and reports the finding of two common RFLPs respectively identified at an RsaI site flanking the 3' end of the gene and at a Hincll site flanking its 5' end. Segregation analysis carried out in the CEPH-pedigrees for the new CDR/RsaI-RFLP versus other polymorphic loci of the region has established a tight linkage with the markers DXS105/DX98 and absence of measurable linkage with two clusters of markers respectively located proximally to the FRAXA (F9, DXS102, DXS51, and DXS369) or distally to it (DXS52, DXS304). In addition, two recombinants were found among 23 scorable sibs identified in the Sardinian pedigrees segregating for the Martin-Bell Syndrome (MBS) and the CDR/RsaI variants. The overall evaluation of the in situ and genetic data reported suggest that the CDR locus 1) is located at the upper boundary of the FRAXA site; 2) is distal to DXS51 and proximal to DXS 389; and 3) segregates in a close linkage association with the loci DXS98 and DXS105 and, to a lesser extent, with the locus for MBS.

Antigens, CD34↗

Vein graft function improvement after percutaneous intervention: evaluation with MR flow mapping.

PURPOSE: To provide functional reference values in single and sequential vein grafts by using magnetic resonance (MR) flow mapping and to examine the effect of percutaneous intervention (PCI) on coronary artery bypass graft function. MATERIALS AND METHODS: Fast MR flow mapping at baseline and during adenosine-induced stress was performed in 39 nonstenotic single vein grafts and 20 nonstenotic sequential vein grafts, as well as in 15 stenotic vein grafts before and 7.3 weeks +/- 1.5 after successful PCI. We evaluated the following parameters (in terms of mean values +/- SDs): average peak velocity (APV) at baseline, stress APV, and velocity reserve. Parameters in nonstenotic single and sequential vein grafts were compared by means of unpaired two-tailed Student t testing. To evaluate changes in velocities before and after PCI, a paired two-tailed Student t test was used. P <.05 was considered to indicate a statistically significant difference. RESULTS: Reference values in single vein grafts for baseline APV, stress APV, and velocity reserve were 8.6 cm/sec +/- 3.4, 20.2 cm/sec +/- 9.5, and 2.4 +/- 0.8, respectively. In sequential vein grafts, significantly higher values for baseline APV (12.2 cm/sec +/- 5.0) and stress APV (27.2 cm/sec +/- 10.6) but a similar velocity reserve (2.3 +/- 0.7) were found. Significant improvements were observed after PCI in baseline APV (before PCI: 9.2 cm/sec +/- 6.6; after PCI: 12.9 cm/sec +/- 7.9; P =.008) and stress APV (before PCI: 12.9 cm/sec +/- 6.3; after PCI: 27.1 cm/sec +/- 13.9; P <.001). No improvement in velocity reserve was observed. CONCLUSION: Significantly higher absolute velocity and flow values were observed in sequential versus single vein grafts, underscoring the need for separate functional reference values for different graft types. Graft function showed significant improvement after PCI to the point that it was restored or nearly restored to reference values.

Adenosine↗

Topography of intensity tuning in cat primary auditory cortex: single-neuron versus multiple-neuron recordings.

1. We studied the spatial distributions of amplitude tuning (monotonicity of rate-level functions) and response threshold of single neurons along the dorsoventral extent of cat primary auditory cortex (AI). To pool data across animals, we used the multiple-unit map of monotonicity as a frame of reference. Amplitude selectivity of multiple units is known to vary systematically along isofrequency contours, which run roughly in the dorsoventral direction. Clusters sharply tuned for intensity (i.e., "nonmonotonic" clusters) are located near the center of the contour. A second nonmonotonic region can be found several millimeters dorsal to the center. We used the locations of these two nonmonotonic regions as reference points to normalize data across animals. Additionally, to compare this study to sharpness of frequency tuning results, we also used multiple-unit bandwidth (BW) maps as references to pool data. 2. The multiple-unit amplitude-related topographies recorded in previous studies were confirmed. Pooled multiple-unit maps closely approximated the previously reported individual case maps when the multiple-unit monotonicity or the map of bandwidth (in octaves) of pure tones to which a cell responds 40 dB above minimum threshold were used as the pooling reference. When the map of bandwidth (in octaves) of pure tones to which a cell responds 10 dB above minimum threshold map was used as part of the measure, the pooled spatial pattern of multiple-unit activity was degraded. 3. Single neurons exhibited nonmonotonic rate-level functions more frequently than multiple units. Although common in single-neuron recordings (28%), strongly nonmonotonic recordings (firing rates reduced by > 50% at high intensities) were uncommon (8%) in multiple-unit recordings. Intermediately nonmonotonic neurons (firing rates reduced between 20% and 50% at high intensities) occurred with nearly equal probability in single-neuron (28%) and multiple-unit (26%) recordings. The remaining recordings for multiple units (66%) and single units (44%) were monotonic (firing rates within 20% of the maximum at the highest tested intensity). 4. In ventral AI (AIv), the topography of monotonicity for single units was qualitatively similar to multiple units, although single units were on average more intensity selective. In dorsal AI (AId) we consistently found a spatial gradient for sharpness of intensity tuning for multiple units; however, for pooled single units in Aid there was no clear topographic gradient. 5. Response (intensity) thresholds of single neurons were not uniformly distributed across the dorsoventral extent of AI.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Human bronchoalveolar lavage fluid: two-dimensional gel electrophoresis, amino acid microsequencing and identification of major proteins.

Although bronchoalveolar lavage has been used as a research and clinical tools for more than two decades to investigate the cellular and soluble components of the lower respiratory tract, its exact protein composition has never been established. In this context, proteins of human bronchoalveolar lavage fluids (BALF), obtained by washing the epithelial lining fluid of the lungs with phosphate-buffered saline, were analyzed by two-dimensional electrophoresis (2-DE) under denaturing and reducing conditions. To characterize the widest amount of proteins, an analytical map of human bronchoalveolar lavage fluid proteins has been created from a pool of BALF from various patients. The resulting map comprises 211 silver-stained spots in the range of pI 3.5-10 and molecular mass 5-100 kDa. We identified 182 spots by microsequence analysis and by matching with human blood plasma and the Macrophage Like Cell line reference 2-DE maps available from the SWISS-2DPAGE database. The human bronchoalveolar lavage fluid was found to contain 61 different proteins or isoforms thereof. Most of the proteins had low molecular masses (< 35 kDa) and rather acidic isoelectric points (pI; 4 < pI < 7). The proteins in the lavage either are produced locally or originate from plasma. Two unknown proteins were identified and are currently under investigation.

Adult↗

Cortical areas within the lateral sulcus connected to cutaneous representations in areas 3b and 1: a revised interpretation of the second somatosensory area in macaque monkeys.

Cortical connections between various body representations in areas 3b and 1 and lateral parietal cortex were examined in 18 macaque monkeys. We injected tracers (Fast Blue, Diamidino Yellow, Horseradish Peroxidase, and Rhodamine Dextran), alone or in combination, into closely related cutaneous responsive sites, e.g., adjacent digits. Separated patches of labeling were found across the parietal operculum and insula for all injected locations. On the basis of cytoarchitectural criteria, the labeled regions include the second somatosensory area (SII), retroinsular area (Ri) and granular insula (Ig). Assuming the connections are homotopical from physiologically identified body representations in primary somatosensory cortex, the labeling patterns in SII include complete anterior and posterior body maps. The orientation of the body is erect in the posterior and supine in the anterior SII region. Area 3b has greater density of connections with anterior SII. The maps are mirror images aligned along the distal extremities. The anterior-posterior (A-P) length of the "SII region" exceeds 7 mm; it extends in the coronal plane from the fundus of the lateral sulcus to surface cortex near the anterior tip of the intraparietal sulcus. Two additional topographically organized maps are likely in Ri. These are "worm-like" body maps oriented along the A-P axis and joined at the head representation. Connections with the center of Ig are not somatotopically organized. The diversity of somatosensory areas in lateral parietal cortex revealed by the labeled connections was discussed in reference to prior mapping of SII in monkeys and was compared to reports of multiple areas in this region of cortex in other species.

Animals↗