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Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly.

The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is finding wide use as a genetic marker that can be directly visualized in the living cells of many heterologous organisms. We have sought to express GFP in the model plant Arabidopsis thaliana, but have found that proper expression of GFP is curtailed due to aberrant mRNA processing. An 84-nt cryptic intron is efficiently recognized and excised from transcripts of the GFP coding sequence. The cryptic intron contains sequences similar to those required for recognition of normal plant introns. We have modified the codon usage of the gfp gene to mutate the intron and to restore proper expression in Arabidopsis. GFP is mainly localized within the nucleoplasm and cytoplasm of transformed Arabidopsis cells and can give rise to high levels of fluorescence, but it proved difficult to efficiently regenerate transgenic plants from such highly fluorescent cells. However, when GFP is targeted to the endoplasmic reticulum, transformed cells regenerate routinely to give highly fluorescent plants. These modified forms of the gfp gene are useful for directly monitoring gene expression and protein localization and dynamics at high resolution, and as a simply scored genetic marker in living plants.

Agrobacterium tumefaciens↗

Carbohydrate recognition systems in autoimmunity.

The immune system is a complex functional network of diverse cells and soluble molecules orchestrating innate and adaptive immunity. Biological information, to run these intricate interactions, is not only stored in protein sequences but also in the structure of the glycan part of the glycoconjugates. The spatially accessible carbohydrate structures that contribute to the cell's glycome are decoded by versatile recognition systems in order to maintain the immune homeostasis of an organism. Microbial carbohydrate structures are recognized by pathogen associated molecular pattern (PAMP) receptors of innate immunity including C-type lectins such as MBL, the tandem-repeat-type macrophage mannose receptor, DC-SIGN or dectin-1 of dendritic cells, certain TLRS or the TCR of NKT cells. Natural autoantibodies, a long known effector branch of this network-based operation, are effective to home in on non-self and self-glycosylation also. The recirculating pool of mammalian immune cells is recruited to inflammatory sites by a reaction pathway involving the self-carbohydrate-binding selectins as initial recognition step. Galectins, further key sensors reading the high-density sugar code, exert regulatory functions on activated T cells, among other activities. Autoimmune diseases are being associated with defined changes of glycosylation. This correlation deserves to be thoroughly studied on the levels of structural mimicry and dysregulation as well as effector molecules to devise innovative anti-inflammatory strategies. This review briefly summarizes data on sensor systems for carbohydrate epitopes and implications for autoimmunity.

Animals↗

Presence of a limited number of essential nucleotides in the promoter region of mouse ribosomal RNA gene.

Point mutations are introduced into a mouse rDNA fragment containing the promoter region by a sodium bisulfite method and the mutants are tested for the ability of accurate transcription initiation in vitro. The results indicate that the change, G to A, at -7 completely eliminates the promoter activity, and those at -16 and at -25 decrease it to about 10% and 50%, respectively. On the other hand, the substitutions at +9, +4, -2, -9 and -39 do not alter the template activity significantly. It is concluded that there are limited but distinct nucleotides that are essential for the transcription initiation of this gene. This sort of absolute requirement for single specific bases is not reported in protein coding genes transcribed by RNA polymerase II. We propose that these rigid recognition signals which we have found are the molecular basis for the strong species-dependency of the transcription machinery of RNA polymerase I system. A model is presented in which a transcription factor interacts with the rDNA promoter from one side of the DNA double-helix with essential contacts at these bases.

Animals↗

The 5' leader of plant PgiC has an intron: the leader shows both the loss and maintenance of constraints compared with introns and exons in the coding region.

PgiC, a complex gene with 23 coding exons and 22 intervening introns, encodes the cytosolic isozyme of phosphoglucose isomerase (EC 5.3.1.9) in higher plants. Here, we report RNA ligase-mediated rapid amplification of cDNA ends experiments that showed that PgiC in Clarkia (Onagraceae) and Arabidopsis thaliana has an intron in the 5' leader. Comparison of the EMBL accessions of the cDNA and genomic sequences showed that this is also the case in rice (Oryza sativa), suggesting that a leader intron is generally present in higher plant PgiC. The intron is bounded by consensus 5'-GT and AG-3' splice sites but showed alternative splicing in Clarkia, resulting in mature transcripts that differ by 8-19 nt in length. The intron is located 18 or 10 nt upstream of the start codon in Clarkia, 2 nt upstream in Arabidopsis, and 9 nt in rice. PgiC in Clarkia was duplicated before the divergence of the extant species, many of which have two expressed genes PgiC1 and PgiC2. Full-length transcripts of both genes identified the transcription start and made it possible to identify the leader intron and leader exon (between the transcription start and leader intron) from previously obtained genomic sequences of both genes in other Clarkia species. These data permit the comparison of evolution in the leader exon and intron with the exons and introns of the coding region, a topic that has not been studied previously. Both the leader exon and the leader intron resemble introns of the coding region in base substitution rate and accumulation of gaps. But the leader intron splice junctions are not strictly conserved in position as are those of the coding region introns. Also, in base composition, the leader intron resembles the other introns, whereas the leader exon more nearly resembles the coding exons. A difference in base composition between coding exons and flanking introns is known to be important for the recognition of splice sites. Thus, the marked difference in base composition between the leader exon and leader intron is probably maintained by selection despite a high rate of sequence divergence.

5' Untranslated Regions↗

Comparison of the F0F2 and F0F1F2 processing strategies for the Cochlear Corporation cochlear implant.

In April 1985, an updated processing strategy became available for the Cochlear Corporation Nucleus cochlear implant. Whereas the original strategy codes only fundamental frequency, amplitude, and information in the second formant region, the newer strategy also codes frequencies in the first formant region. This investigation evaluated the speech recognition skills of five subjects who were experienced with both designs. On average, the addition of first formant information improved word identification in an audition-only condition and improved spondee recognition in noise. Scores for the NU 6 Monosyllabic Word Test and the Sentence Test Without Context improved from 8% (2-12%) to 28% words correct (10-42%), and from 31% (10-45%) to 64% words correct (39-84%), respectively. Scores for the Four-Choice Spondee Test in noise improved from 37% (25-50%) to 75% (45-90%). The percent correct scores for the Iowa 14-Item Consonant Confusion Test in an audition-only and vision-plus-audition condition did not vary with the change in processing strategy. However, an information transfer analysis performed on the responses to the consonant test in a vision-only and a vision-plus-audition condition suggested that the newer strategy enhances the transmission of the voicing, duration, and envelope features.

Adult↗

Specificity of multiple homologous genomic regions in Spodoptera exigua nucleopolyhedrovirus DNA replication.

The region upstream of the Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV) ubiquitin gene contains four near-identical 68-bp-long palindromic repeats. This region, named Sehr6 and located at map unit (m.u.) 88 of the SeMNPV genome on pSeEcoRI-2.2, showed structural homology to previously identified homologous regions (hrs) in a number of other baculoviruses. Hrs function as enhancers of transcription and as putative origins (oris) of baculovirus DNA replication. Five additional hrs (Sehr1-Sehr5) were identified on the SeMNPV genome by Southern blot hybridization with an 18-bp-long oligonucleotide complementary to a sequence conserved within the arms of the four palindromic repeats of Sehr6. Sehr1-Sehr6 were dispersed on the SeMNPV genome at m.u. 8.0, 30.0, 38.5, 51.0, 77.0 and 88.0, respectively. Sequence analysis of these hrs confirmed the presence of palindromic repeats, highly similar to those found in pSeEcoRI-2.2. The number of palindromes varied from one (Sehr4) to nine (Sehr1) per hr. The Sehrs are all present in non-coding regions of the SeMNPV genome and also contain multiple putative transcription recognition sequences. Plasmids containing either of the Sehrs replicated in an SeMNPV-dependent DNA replication assay. The Sehrs were unable to replicate in an AcMNPV-dependent DNA replication assay. This was in contrast to the previously observed SeMNPV non-hr type ori, which replicated in the presence of both AcMNPV and SeMNPV. These data suggest that the replication of SeMNPV and the role of hrs in this process is highly specific.

Animals↗

Concept hierarchy memory model: a neural architecture for conceptual knowledge representation, learning, and commonsense reasoning.

This article introduces a neural network based cognitive architecture termed Concept Hierarchy Memory Model (CHMM) for conceptual knowledge representation and commonsense reasoning. CHMM is composed of two subnetworks: a Concept Formation Network (CFN), that acquires concepts based on their sensory representations; and a Concept Hierarchy Network (CHN), that encodes hierarchical relationships between concepts. Based on Adaptive Resonance Associative Map (ARAM), a supervised Adaptive Resonance Theory (ART) model, CHMM provides a systematic treatment for concept formation and organization of a concept hierarchy. Specifically, a concept can be learned by sampling activities across multiple sensory fields. By chunking relations between concepts as cognitive codes, a concept hierarchy can be learned/modified through experience. Also, fuzzy relations between concepts can now be represented in terms of the weights on the links connecting them. Using a unified inferencing mechanism based on code firing, CHMM performs an important class of commonsense reasoning, including concept recognition and property inheritance.

Algorithms↗

Genes for control of spread of the tumor and Mycobacterium tuberculosis infection in the mouse.

In this review, new data are interpreted that provide some answers to the question of why the immune system fails to respond to metastatic cancers. A function such as an element of the immune response is expected to be under the control of a gene. To find a gene, a mutant is required. Complex screens have been designed and used to detect mouse mutants resisting spread of transplantable tumors (in a spontaneous metastasis assay). Curiously, both mutants with increased and decreased resistance to spread of the tumor have been found. S-27 is a strongly resistant mutant linked to MHC; this mutation also affected some responses to Mycobacterium tuberculosis infection. A long sought link between malignant transformation and vaccination against mycobacterial diseases is discovered in this mutant. A search for a molecule responsible for effects of the S-27 mutation resulted in an unexpected finding. The S-27 mutant carries complex nucleotide alterations at the junction between the signal sequence and the sequences coding for the mature MHC class II A beta polypeptide chain. Antigen recognition region of the A beta molecule is not affected by this mutation. Some concepts developed during the course of this study are illustrated in Figures 1 to 4.

Amino Acid Sequence↗

Perceptual defense mechanisms in Crohn's disease and panic disorder.

The defense organization of Crohn's disease and Panic Disorder was studied with a well-validated tachistoscopic paradigm, the Defense Mechanism Test. Three sex- and age-matched groups of 34 subjects (Crohn, Panic, and Nonclinical) were compared on the main codings of defense. Crohn patients were characterized by stereotypy and lack of recognition of the threat. The Panic group presented clearly higher frequencies of protocols with repression, isolation, and disappearance of the threat. Especially was the strongest variant of isolation, barrier-isolation, typical of the defensive substructure of panic patients.

Adult↗

Phonological and orthographic coding skills in adult readers.

Although less skilled readers are handicapped by their poor phonological skills, this may not be true of their visual and orthographic coding skills. Because of an increasing reliance on visual-orthographic coding with reading experience, the author predicted that there would be smaller differences between skilled and less skilled adult readers on orthographic coding measures than on phonological coding measures. The orthographic and phonological coding measures involved, respectively, judgments of which looks more like a word, filv-fild, and which sounds like a real word, kake-dake? On the orthographic measure, reading groups did not differ in coding speed--although the less skilled readers made more errors, but far fewer than on the phonological coding measure. Differences between reading groups were substantial for both speed and errors on the phonological coding measure. Phonological variables accounted for most of the variance in word recognition, and this was especially true for men. The results suggest that in less skilled adult readers, phonological skills are a primary factor in their reading despite some evidence of visual-orthographic compensation.

Adolescent↗

Recognition memory as a function of encoding strategy and stimulus codability.

The primary purpose of this research was to determine how imaginal and verbal encoding strategies intereact with various stimulus characteristics to either enhance or retard recognition; the secondary purpose of these studies was to test the conceptual coding hypothesis of Ellis, 1972. A between-groups multivariate factorial analysis of covariance experiment and a within-subjects multivariate factorial analysis of variance experiment were conducted. In Experiment 1 it was found that low-codability shapes were better recognized under the verbal encoding set rather than the imaginal encoding verbal encoding sets; and high-codability shapes were not better recognized than low-codability shapes. However, in Experiment 2, where instructional set was a within-subjects factor, it was found that low-codability shapes were not better recognized under the verbal encoding set than the imaginal encoding set.

Adult↗

Implicit multisensory associations influence voice recognition.

Natural objects provide partially redundant information to the brain through different sensory modalities. For example, voices and faces both give information about the speech content, age, and gender of a person. Thanks to this redundancy, multimodal recognition is fast, robust, and automatic. In unimodal perception, however, only part of the information about an object is available. Here, we addressed whether, even under conditions of unimodal sensory input, crossmodal neural circuits that have been shaped by previous associative learning become activated and underpin a performance benefit. We measured brain activity with functional magnetic resonance imaging before, while, and after participants learned to associate either sensory redundant stimuli, i.e. voices and faces, or arbitrary multimodal combinations, i.e. voices and written names, ring tones, and cell phones or brand names of these cell phones. After learning, participants were better at recognizing unimodal auditory voices that had been paired with faces than those paired with written names, and association of voices with faces resulted in an increased functional coupling between voice and face areas. No such effects were observed for ring tones that had been paired with cell phones or names. These findings demonstrate that brief exposure to ecologically valid and sensory redundant stimulus pairs, such as voices and faces, induces specific multisensory associations. Consistent with predictive coding theories, associative representations become thereafter available for unimodal perception and facilitate object recognition. These data suggest that for natural objects effective predictive signals can be generated across sensory systems and proceed by optimization of functional connectivity between specialized cortical sensory modules.

Acoustic Stimulation↗

The effect of word concreteness on recognition memory.

Concrete words that are readily imagined are better remembered than abstract words. Theoretical explanations for this effect either claim a dual coding of concrete words in the form of both a verbal and a sensory code (dual-coding theory), or a more accessible semantic network for concrete words than for abstract words (context-availability theory). However, the neural mechanisms of improved memory for concrete versus abstract words are poorly understood. Here, we investigated the processing of concrete and abstract words during encoding and retrieval in a recognition memory task using event-related functional magnetic resonance imaging (fMRI). As predicted, memory performance was significantly better for concrete words than for abstract words. Abstract words elicited stronger activations of the left inferior frontal cortex both during encoding and recognition than did concrete words. Stronger activation of this area was also associated with successful encoding for both abstract and concrete words. Concrete words elicited stronger activations bilaterally in the posterior inferior parietal lobe during recognition. The left parietal activation was associated with correct identification of old stimuli. The anterior precuneus, left cerebellar hemisphere and the posterior and anterior cingulate cortex showed activations both for successful recognition of concrete words and for online processing of concrete words during encoding. Additionally, we observed a correlation across subjects between brain activity in the left anterior fusiform gyrus and hippocampus during recognition of learned words and the strength of the concreteness effect. These findings support the idea of specific brain processes for concrete words, which are reactivated during successful recognition.

Adult↗

Should there be a separate code of nomenclature for the protists?

The present Botanical and Zoological Codes of Nomenclature are often inadequate for resolution of all the peculiar problems caused by the very nature of the numerous and diverse groups of the so-called 'lower' eukaryotic organisms known as protists. Whether or not a separate code should therefore be created for these species--many but not all of which are unicellular in structure and microscopic in size--is complicated by several factors. The principal one is related to the wide dispersal of protists throughout many taxonomic classes and phyla/divisions; sometimes even multiple kingdoms are involved. If recognition of a single kingdom Protista is no longer tenable, then even the concept of one code per kingdom is not applicable. Other difficulties arise primarily from long-standing differences in major provisions of present Botanical and Zoological Codes. Numerous 'ambiregnal' forms exist, species currently under dual code jurisdiction. The matter of names for suprafamilial taxa of protists, irrespective of their ultimate kingdom assignment, poses another set of concerns not yet resolved. A plea is made to recognize the legitimacy of having distinct high-level ranks for protist species that seem to be widely separated phylogenetically from fellow protists or from other eukaryotic assemblages.

Animals↗

Altruism through beard chromodynamics.

The evolution of altruism, a behaviour that benefits others at one's own fitness expense, poses a darwinian paradox. The paradox is resolved if many interactions are with related individuals so that the benefits of altruism are reaped by copies of the altruistic gene in other individuals, a mechanism called kin selection. However, recognition of altruists could provide an alternative route towards the evolution of altruism. Arguably the simplest recognition system is a conspicuous, heritable tag, such as a green beard. Despite the fact that such genes have been reported, the 'green beard effect' has often been dismissed because it is unlikely that a single gene can code for altruism and a recognizable tag. Here we model the green beard effect and find that if recognition and altruism are always inherited together, the dynamics are highly unstable, leading to the loss of altruism. In contrast, if the effect is caused by loosely coupled separate genes, altruism is facilitated through beard chromodynamics in which many beard colours co-occur. This allows altruism to persist even in weakly structured populations and implies that the green beard effect, in the form of a fluid association of altruistic traits with a recognition tag, can be much more prevalent than hitherto assumed.

Altruism↗

H-2 restriction of cell-mediated immunity to an intracellular bacterium: effector T cells are specific for Listeria antigen in association with H-21 region-coded self-markers.

The protective activity of anti-Listeria-immune T cells assayed in an adoptive transfer system in H-2 restricted. As shown in the present studies, the demonstration of the restriction is directly dependent on the dose and the relative protective activity of spleen cells. In addition, some H-2-unrestricted protection is conferred predominantly by other than immunoglobulin-negative spleen cells. Thus, the activity of Listeria-immune T cells appears to be 'absolutely' restricted and is in this respect comparable to in vivo T-cell-mediated anti-viral protection. The predominant genetic region of H-2 coding for the structures which are mainly involved in this restriction in T-cell immunity to this prototype intracellular bacterium is the I region. The specificity of Listeria-immune T cells is determined by the H-2 haplotype of the donor. Thus, F1 hybrids seem to possess at least two separable sets of T cells, each specific for one parental haplotype. As is true in the virus model, the results cannot distinguish between an altered-self or a dual recognition model of T-cell recognition to explain H-2 restriction. They are, however, compatible with the idea and I-coded cell surface structures may serve as receptors for cell-specific differentiation signals, which trigger direct or lymphokin-mediated activation of macrophages to manifest increased bactericidal capacity. The interesting parallels in self-marker recognition of T cells in the virus and intracellular bacterium systems, respectively, appear to be reasonably explained by the different types of signals transmitted by T cells to various target cells via the distinctly different self-markers employed (i.e., K or D vs I).

Animals↗

The IPRS Image Processing and Pattern Recognition System.

IPRS is a freely available software system which consists of about 250 library functions in C, and a set of application programs. It is designed to run under UNIX and comes with full source code, system manual pages, and a comprehensive user's and programmer's guide. It is intended for use by researchers in human vision, pattern recognition, image processing, machine vision and machine learning.

Humans↗

The cDNA for the beta-subunit of human chorionic gonadotropin suggests evolution of a gene by readthrough into the 3'-untranslated region.

A 579-base pair approximately full-length cDNA which codes for the 145-amino acid long beta-subunit of human chorionic gonadotropin (HCG) has been cloned in the plasmid vector pBR322 and its complete nucleotide sequence determined. A hydrophobic presequence of 20 amino acids can be identified from the nucleotide sequence. The amino acid sequence of the beta-subunit is known to be related to those of the beta-subunits of the other glycoprotein hormones LH, FSH and TSH, but the beta-subunit of HCG is unique in that it contains a C-terminal extension of about 30 amino acids which has no homologous counterpart in the other three hormones. Analysis of the beta HCG cDNA nucleotide sequence suggests that this extension may have arisen by the loss of the termination codon of an ancestral beta-like gene so that most of what was previously the 3'-untranslated region now codes for protein. The beta-subunit of HCG terminates with the codon UAA located 16 bases before the poly(A) in the sequence AAUAAA. This sequence is believed to be a recognition signal involved in either polyadenylation or processing and therefore has a dual role in this gene, serving both a coding and a regulatory function.

Base Sequence↗