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The two-domain hypothesis in Beckwith-Wiedemann syndrome: autonomous imprinting of the telomeric domain of the distal chromosome 7 cluster.

A large cluster of imprinted genes is located on the mouse distal chromosome 7. This cluster is well conserved in humans and its dysregulation results in the overgrowth- and tumour-associated Beckwith-Wiedemann syndrome. Two imprinting centres (IC1 and IC2) controlling different sets of genes have been identified in the cluster, raising the hypothesis that the cluster is divided into two functionally independent domains. However, the mechanisms by which imprinting of genes in the IC2 domain (e.g. Cdkn1c and Kcnq1) is regulated have not been well defined, and recent evidence indicates that distantly located cis-acting elements are required for IC2 imprinting. We show that the maternal germ-line methylation at IC2 and the imprinted expression of five genes of the IC2 domain are correctly reproduced on an 800 kb YAC transgene when transferred outside of their normal chromosomal context. These results, together with previous transgenic studies, locate key imprinting control elements within a 400 kb region centromeric of IC2 and demonstrate that each of the two domains of the cluster contains the cis-acting elements required for the imprinting control of its own genes. Finally, maternal, but not paternal, transmission of the transgene results in fetal growth restriction, suggesting that during evolution the acquisition of imprinting may have been facilitated by the opposite effects of the two domains on embryo growth.

Animals↗

The structure of the rat amiloride-sensitive epithelial sodium channel gamma subunit gene and functional analysis of its promoter.

Prolonged dietary Na+ depletion and chronic administration of adrenal steroids increase steady-state mRNA levels of the gamma subunit of the epithelial sodium channel (gammaENaC) in rat colon. This increase correlates with a marked increase in transepithelial Na+ transport and is thought to occur via transcriptional regulation. To begin to evaluate these mechanisms in detail, we determined the organization of the rat gammaENaC gene. A rat genomic library was screened and overlapping lambda clones that together span the gene (approximately 36 kb) and contain at least 1 kb of 5' flanking genomic DNA were isolated. As in the human gene, the rat gammaENaC gene contains 13 exons and a CpG island at the 5' end of the gene. A single transcription start site was identified in rat kidney by nuclease protection assay defining a 5' untranslated region of 126 nt. The translation initiation codon was identified within the second exon and the entire 3' UTR (approximately 1 kb) was within the last exon. 800 bp of 5' flanking sequence, as well as the 3.4 kb first intron, were sequenced and analyzed for transcriptional regulatory motifs. Analogous to the human gammaENaC gene [Thomas, C.P., Doggett, N.A., Fisher, R., Stokes J.B., 1996. Genomic organization and the 5' flanking region of the gamma subunit of the human amiloride-sensitive epithelial sodium channel. J. Biol. Chem. 271, 26 062--26 066], two GC boxes were seen at -30 and -61 to the transcription start site. In addition, putative AP-1, AP-2, CRE, Sp1 and GATA-1 and GRE motifs were identified elsewhere in the 5' flanking region or the first intron. Two mammalian-wide interspersed repeats and a rodent-specific B1 repeat were also identified within the first intron. Fragments containing the putative GRE motifs coupled to luciferase did not confer a glucocorticoid-stimulated response in two cell lines that contained a functional glucocorticoid receptor. However, a 76 nt rat gammaENaC 5' flanking fragment (-76 to +68) directed expression of luciferase in the epithelial cell lines H441 and FRTL5, suggesting that this minimal region that contained both GC boxes was sufficient for promoter activity.

Animals↗

Sequence-based exon prediction around the synaptophysin locus reveals a gene-rich area containing novel genes in human proximal Xp.

The human Xp11.23-p11.22 interval has been implicated in several inherited diseases including Wiskott-Aldrich syndrome; three forms of X-linked hypercalciuric nephrolithiaisis; and the eye disorders retinitis pigmentosa 2, congenital stationary night blindness, and Aland Island eye disease. In constructing YAC contigs spanning Xp11. 23-p11.22, we have previously shown that the region around the synaptophysin (SYP) gene is refractory to cloning in YACs, but highly stable in cosmids. Preliminary analysis of the latter suggested that this might reflect a high density of coding sequences and we therefore undertook the complete sequencing of a SYP-containing cosmid. Sequence data were extensively analyzed using computer programs such as CENSOR (to mask repeats), BLAST (for homology searches), and GRAIL and GENE-ID (to predict exons). This revealed the presence of 29 putative exons, organized into three genes, in addition to the 7 exons of the complete SYP coding region, all mapping within a 44-kb interval. Two genes are novel, one (CACNA1F) showing high homology to alpha1 subunits of calcium channels, the other (LMO6) encoding a product with significant similarity to LIM-domain proteins. RT-PCR and Northern blot studies confirmed that these loci are indeed transcribed. The third locus is the previously described, but not previously localized, A4 differentiation-dependent gene. Given that the intron-exon boundaries predicted by the analysis are consistent with previous information where available, we have been able to suggest the genomic organization of the novel genes with some confidence. The region has an elevated GC content (>53%), and we identified CpG islands associated with the 5' ends of SYP, A4, and LMO6. The order of loci was Xpter-A4-LMO6-SYP-CACNA1F-Xcen, with intergenic distances ranging from approximately 300 bp to approximately 5 kb. The density of transcribed sequences in this area (>80%) is comparable to that found in the highly gene-rich chromosomal band Xq28. Further studies may aid our understanding of the long-range organization surrounding such gene-enriched regions.

Amino Acid Sequence↗

Esp-independent biofilm formation by Enterococcus faecalis.

Enterococcus faecalis is a gram-positive opportunistic pathogen known to form biofilms in vitro. In addition, this organism is often isolated from biofilms on the surfaces of various indwelling medical devices. However, the molecular mechanisms regulating biofilm formation in these clinical isolates are largely unknown. Recent work has suggested that a specific cell surface protein (Esp) of E. faecalis is critical for biofilm formation by this organism. However, in the same study, esp-deficient strains of E. faecalis were found to be capable of biofilm formation. To test the hypothesis that Esp is dispensable for biofilm formation by E. faecalis, we used microtiter plate assays and a chemostat-based biofilm fermentor assay to examine biofilm formation by genetically well-defined, non-Esp-expressing strains. Our results demonstrate that in vitro biofilm formation occurs, not only in the absence of esp, but also in the absence of the entire pathogenicity island that harbors the esp coding sequence. Using scanning electron microscopy to evaluate biofilms of E. faecalis OG1RF grown in the fermentor system, biofilm development was observed to progress through multiple stages, including attachment of individual cells to the substratum, microcolony formation, and maturation into complex multilayered structures apparently containing water channels. Microtiter plate biofilm analyses indicated that biofilm formation or maintenance was modulated by environmental conditions. Furthermore, our results demonstrate that expression of a secreted metalloprotease, GelE, enhances biofilm formation by E. faecalis. In summary, E. faecalis forms complex biofilms by a process that is sensitive to environmental conditions and does not require the Esp surface protein.

Bacterial Proteins↗

Two gene determinants are differentially involved in the biogenesis of Fap1 precursors in Streptococcus parasanguis.

Mature Fap1, a 200-kDa fimbria-associated adhesin, is required for fimbrial biogenesis and biofilm formation in Streptococcus parasanguis. Fap1-like proteins are found in the genomes of many streptococcal and staphylococcal species. Fap1 is a serine-rich glycoprotein modified by O-linked glycan moieties. In this study, we identified a seven-gene cluster including secY2, orf1, orf2, orf3, secA2, gtf1, and gtf2 that is localized immediately downstream of fap1. The lower G+C contents and the presence of a putative transposase element suggest that this gene cluster was horizontally transferred from other bacteria and represents a genomic island. At least two genes in this island mediated Fap1 biogenesis. Mutation of a glucosyltransferase (Gtf1) gene led to accumulation of a Fap1 precursor, which had no detectable glycan moieties. Inactivation of a gene coding for an accessory Sec protein (SecY2) resulted in expression of a distinct Fap1 precursor, which reacted with one glycan-specific Fap1 antibody but not with another glycan-specific antibody. Furthermore, partially glycosylated Fap1 was detected on the cell surface and in the culture supernatant. These data suggest that SecY2 has a role in complete glycosylation of Fap1 and imply that SecY2 is not the only translocation channel for the Fap1 precursor and that alternative secretion machinery exists. Together, Gtf1 and SecY2 are involved in biogenesis of two distinct Fap1 precursors in S. parasanguis. Discovery of the effect of an accessory Sec protein on Fap1 glycosylation suggests that Fap1 secretion and glycosylation are coupled during Fap1 biogenesis.

Bacterial Proteins↗

Metatropic dwarfism. Uncoupling of endochondral and perichondral growth.

Metatropic dwarfism is a rare heritable skeletal dysplasia that is thought to result from a defect in endochondral ossification. Histological studies have been few and have yielded inconsistent findings. In addition, no investigator has commented on the structure and function of the perichondral portion of the growth plate in patients who have metatropic dysplasia. To further characterize this disturbance, histological studies were carried out on autopsy specimens from the proximal part of the femur and the iliac crest of a patient who had this disorder. The major findings were: the absence of formation of normal primary spongiosa in the metaphysis; the presence of a thin seal of bone at the chondro-osseous junction, with abnormal metaphyseal vascular invasion and arrest of endochondral growth; and normal-appearing perichondral ring structures with persistence of circumferential growth. These findings suggest an uncoupling of endochondral and perichondral growth and offer an explanation for the dumbbell-shaped morphological structure of the osseous metaphysis that is seen in patients who have metatropic dysplasia. Other observations included prominence of the cartilaginous canals and vascular channels in the reserve zone; clumping of chondrocytes with enhanced staining of the pericellular matrix in the proliferative zone; a decreased ratio of cells to matrix in the hypertrophic zone, with intracellular metachromatic granules and incomplete evolution of chondrocytes; complete absence of an alcian-blue-positive zone of provisional calcification; and, finally, islands of dysplastic chondrocytes in the metaphysis. These abnormalities suggest that metatropic dysplasia is not simply a disorder of endochondral ossification. There appear to be associated defects in the longitudinal proliferation and maturation of chondrocytes and in the production of normal matrix.

Achondroplasia↗

Presence of calcium antagonist activity explains the use of Syzygium samarangense in diarrhoea.

Syzygium samarangense (Family; Myrtaceae) or 'makopa', as it is commonly known, is native to Malaysia, some islands of Indonesia and to Far East in general. This study was undertaken to rationalize the use of this plant in hypermotility states of the gut. The hexane extract of S. samarangense (Ss.Hex) was found to dose-dependently (10-3000 microg/mL) relax the spontaneously contracting isolated rabbit jejunum. When tested for a possible calcium channel blocking (CCB) activity, the extract (10-1000 microg/mL) relaxed the high K+-induced contractions and also decreased the Ca++ dose-response curves in a dose-dependent manner (30-100 microg/mL), confirming the CCB activity. Four flavonoids isolated from the hexane extract were tested for a possible spasmolytic activity. All flavonoids, identified as: 2'-hydroxy-4',6'-dimethoxy-3'-methylchalcone (SS1), 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (SS2), 2',4'-dihydroxy-6'-methoxy-3'-methylchalcone (SS3) and 7-hydroxy-5-methoxy-6,8-dimethylflavanone (SS4), showed dose-dependent (10-1000 microg/mL) spasmolytic activity with SS2 being the most potent. These results indicate that the presence of compounds with spasmolytic and calcium antagonist activity may be responsible for the medicinal use of the plant in diarrhoea.

Animals↗

Intestinal electrolyte secretion. History of a paradigm.

The contemporary paradigm for active chloride secretion by vertebrate epithelial cells evolved, at least in part, from experiments that began in the laboratory of Dr William Silen at Beth Israel Hospital in Boston, Mass. It was first shown there that cyclic adenosine monophosphate and cholera toxin stimulate active chloride secretion when added to intestinal mucosa in vitro. The paradigm, which evolved further from experiments on shark rectal gland and flounder intestine at the Mt Desert Island Biological Laboratory in Salsbury Cove, Maine, is as follows: Chloride enters some epithelial cells by sodium-potassium-chloride cotransport with a stoichiometry of 1:1:2 and accumulates intracellularly because of the sodium gradient maintained by sodium-potassium-adenosinetriphosphatase in the basolateral membrane; the chloride is then released from the cell through chloride channels in the membrane opposite that of the cotransporter. If the cotransporter is basolateral and the channel is apical, chloride is secreted; if it is the other way around, chloride is absorbed. In a number of secretory epithelial cells, cyclic adenosine monophosphate activates these channels, thereby initiating secretion. A defect in the activation of these channels by cyclic adenosine monophosphate is the root cause of cystic fibrosis.

Animals↗

Discordance between malignant hyperthermia susceptibility and RYR1 mutation C1840T in two Scandinavian MH families exhibiting this mutation.

The ryanodine receptor 1 (RYR1) mutation C1840T has been reported to segregate with malignant hyperthermia (MH) susceptibility in several families. We have investigated several Scandinavian MH families with respect to five different RYR1 mutations reported to cause predisposition to MH, and we here report on two of the families in which the C1840T mutation was detected. In these two families there was recombination between MH susceptibility and this mutation in one and three individuals, respectively. These findings may suggest that it is necessary to reconsider the specificity of the IVCT and the role of C1840T as a cause of MH susceptibility in some families exhibiting this mutation.

Anesthesia↗

Utilization of military support in the response to hurricane Marilyn: implications for future military-civilian cooperation.

INTRODUCTION: The past decade has been a period of evolution for the Federal disaster response system within the United States. Two domestic hurricanes were pivotal events that influenced the methods used for organizing Federal disaster assistance. The lessons of Hurricane Hugo (1989) and Hurricane Andrew (1992) were incorporated into the successful response to Hurricane Marilyn in the U.S. Virgin Islands in 1995. Following each of these storms, the Department of Defense was a major component of the response by the health sector. Despite progress in many areas, lack of clear communication between military and civilian managers and confusion among those requesting Department of Defense health resources may remain as obstacles to rapid response. METHODS: This discussion is based on an unpublished case report utilizing interviews with military and civilian managers involved in the Hurricane Marilyn response. RESULTS: The findings suggest that out-of-channel pathways normally utilized in the warning and emergency phase of the response remained operational after more formal civilian-military communication pathways and local assessment capability had been established. CONCLUSION: It is concluded that delays may be avoided if the system in place was to make all active pathways for the request and validation of military resources visible to the designated Federal managers located within the area of operations.

Attitude of Health Personnel↗

Differential nicotinic acetylcholine receptor subunit expression in the human hippocampus.

Neuronal nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels composed of alpha and beta subunits with specific structural, functional and pharmacological properties. In this study the distribution of alpha3, alpha4, alpha7, beta2 and beta4 nAChR subunits in the human hippocampus was investigated using immunohistochemistry. Most pyramidal neurons, pre-alpha cells of the entorhinal cortex and dentate granule cells were immunoreactive for all subunits. Small islands of alpha7 immunoreactive cells were present in the outer presubiculum. alpha4 and beta2, and alpha3, alpha4 and beta2 immunoreactive fibre tracts were present in the stratum radiatum and subiculum, respectively, suggesting nAChRs may play a role in modulating inputs to the hippocampus via Schaffer collaterals and along the perforant pathway. Some astrocytes were immunoreactive for alpha3, alpha7 and beta4 subunits. Immunoreactivity to all subunits was noted in association with blood vessels. These results indicate the involvement of multiple nAChR subtypes in the modulation of both neuronal and non-neuronal functions in the human hippocampus.

Adult↗

ENVIRONMENTAL AUDITING: The Fish Fauna of the Doubs River Prior to Completion of the Rhine-Rhone Connection

/ The part of the Doubs River between Montbeliard and Dole (France), i.e., downstream from the confluence with the Allan River, will be affected by the Rhine- Rhone connection project. In order to improve the understanding of the Doubs ichthyofauna, aquatic environments of the Doubs were sampled by electrofishing. Fish diversity and the presence of some rheophilic species demonstrated the good ecological quality of some stretches of the Doubs. This quality was due to alternating areas with very diversified aquatic environments (riffles, islands and side-arms, backwaters) and a considerable range of flow velocities. The differences in the structure of the fish communities of the different types of aquatic environments were more qualitative (fish species) than quantitative (number of species and number of fish). However, the mean number of fish was statistically lower in the canals (Freycinet canal and channelized part of the Allan River) than in the main course and in the backwaters. The natural parts of the Doubs (unnavigable reaches) showed the most diversified environmental structure and had the most rheophilic fish communities. Thus, the rheophilic species were well represented, but they proved also the most vulnerable to river regulation. However, the most abundant fishes throughout the Doubs River were generalists with no special requirements for food sources or spawning substrate.KEY WORDS: Fish communities; Regulation; Restoration; Floodplain; Large ship canal; Doubs River

Journal Article↗

A study of the saphenous venous island flap in the dog without arterial inflow using a non-biological conduit across a part of the length of the vein.

This paper includes material which is a follow-up on the work of Se Min Baek et al. (1985) who demonstrated that in dogs a saphenous flap could survive without arterial inflow. Here we have repeated part of their work and confirmed it. In addition, this paper shows that in dogs a saphenous venous flap with only the cephalad venous channel intact can survive even if the vein is cut and replaced by a non-biological conduit (a polyethylene tube). A total of 13 dogs and 19 flaps were studied. The possible reasons for the survival of these flaps are discussed.

Animals↗

The use of data loggers to determine the energetics and physiology of aquatic birds and mammals.

By deploying a data logger specifically designed for the purpose, it was possible to record heart rate, fH (beats/min), from free-ranging gentoo penguins, Pygoscelis papua, and Antarctic fur seals, Arctocephalus gazella, at the British Antarctic Survey Base at Bird Island. The heart rate data were then converted into oxygen consumption (VO2, mlO2 min-1 kg-1) and/or energy expenditure (W/kg) using equations that had been derived from calibration experiments. In these experiments the relationships between fH and VO2 were determined in animals at rest and while exercising at different levels on a treadmill or in a static water channel (penguins) and in a variable speed flume (California sea lions, Zalophus californianus, as surrogate fur seals) or in a static water channel (fur seals). The validity of using these relationships was tested by recording simultaneously fH, VO2 by direct respirometry and VCO2 by the doubly labelled water (DLW) technique in six penguins and in six California sea lions during 72 h and 24 h, respectively, at various levels of activity. For the penguins, both indirect methods gave mean algebraic errors within 2% of the measured VO2, whereas for the sea lions, the mean algebraic errors were 36.4% for the DLW method and 2.7% for the fH method. The range of errors was greater for the DLW method, in both species. Field data from 15 penguins indicate that the fH method provides data that are comparable to those obtained using the DLW method, but with the added advantage that they can be broken down into the energy costs for specific types of behaviour. The implanted data loggers also recorded the temperature of the abdominal cavity (T(ab)) and it was evident that this routinely decreased by approximately 2 degrees C during diving bouts and by a maximum of almost 5 degrees C. Such temperature decreases, particularly if representative of similar decreases in other tissues, may at least partly explain why the energy costs of travelling to and from the foraging site and of foraging itself are similar to those for penguins resting in water at 5 degrees C. Field data from 15 female fur seals indicate that, when the animals are ashore, there is good agreement between the values for mean energy expenditure obtained by the fH and DLW methods. However, when the animals are at sea, the values obtained by DLW are substantially greater than those obtained by fH. When the at-sea values are corrected for the apparent overestimations referred to above, there is good agreement between the data obtained using the two methods. The data derived from fH indicate that, as with the penguins, the energy expenditures during travelling to and from the foraging site and during foraging are similar to those of fur seals resting in water at 7 degrees C.

Animals↗

Identification of the alternative promoters of the KChIP4 subfamily.

The subfamily of voltage-dependent potassium (Kv) channel interacting protein 4 (KChIP4) is made up of the auxiliary interacting protein of voltage-dependent potassium channels. In this study, the structure of four splicing variants of the human KChIP4 gene was analyzed. Three of the four isoforms of the KChIP4 gene, KChIP4.1, KChIP4.2 and KChIP4.4, were amplified from mouse and human fetal brain tissues by reverse transcription-polymerase chain reaction and then identified. Based on the bioinformatics analysis of the genomic sequences of the gene, we cloned and characterized two promoter fragments from the KChIP4 gene. One was a 325 bp fragment upstream of the 5' end of the KChIP4.1 mRNA sequence and the other was an 818 bp fragment located immediately at the 5' end of the KChIP4.4 variant. Both of them can initiate the transcription of the reporter gene in HT1080 cells and Sprague-Dawley (SD) rat fetal brain neurons, and they contain CG islands, except typical TATA boxes and CAAT boxes. This shows that the KChIP4 gene expression is regulated by an alternative promoter.

Amino Acid Sequence↗

Epigenetic profiling of synchronous colorectal neoplasias by quantitative DNA methylation analysis.

Transcriptional inactivation of tumor-suppressor genes by promoter CpG island methylation is thought to be an important mechanism in human carcinogenesis. The CpG island methylator phenotype (CIMP) with extensive promoter methylation appears to be a distinct epigenetic subtype of colorectal carcinoma. Most previous studies on CpG island methylation in colorectal carcinoma used methylation-specific PCR, which may detect low levels of DNA methylation with little or no biological significance. In contrast, quantitative DNA methylation assays have been shown to provide useful information beyond that which can be achieved with methylation-specific PCR. Synchronous neoplasias provide a unique model for investigators to examine molecular alterations in multistep tumorigenesis within one individual. However, no study to date has quantified DNA methylation of CIMP-specific promoters in synchronous colorectal neoplasias. Utilizing real-time PCR (MethyLight), we quantified DNA methylation in five CIMP-specific gene promoters [CACNA1G (calcium channel, voltage-dependent, T type alpha-1G subunit), CDKN2A (p16/INK4A), CRABP1 (cellular retinoic acid binding protein-1), MLH1 and NEUROG1 (neurogenin 1)] and MGMT in six synchronous carcinoma pairs (12 carcinomas) and eight synchronous carcinoma and adenoma pairs (16 tumors). We found that while some synchronous tumor pairs showed discordant promoter methylation patterns, other tumor pairs showed similar, but not exactly identical, patterns of promoter methylation. All but two pairs showed concordant patterns of CIMP status (CIMP positive vs CIMP negative) (P = 0.05 in cancer pairs). BRAF mutations were present in only CIMP-positive tumors. A high degree of microsatellite instability (MSI-H) was observed in both CIMP-positive and CIMP-negative tumors. KRAS mutations were not concordant in any synchronous neoplasia pair. In conclusion, epigenetic alterations at CIMP-specific promoter CpG islands in synchronous colorectal neoplasias likely have both random and nonrandom components.

Adenocarcinoma↗

Distribution of mRNAs coding for liver and heart gap junction proteins in the rat central nervous system.

The present study examined the distributions of connexin43 mRNA and connexin32 mRNA in the central nervous system (CNS) of the rat by using in situ hybridization histochemistry. These connexins are the best studied gap junction proteins; connexin32 forms direct cell-cell channels in the liver, as does connexin43 in the heart. There was a differential distribution of cells containing connexin32 mRNA compared with the population of cells which contained connexin43 mRNA, thus implying a regional specificity in the expression of connexins in the CNS. Cells containing connexin43 mRNA were uniformly distributed throughout the gray matter of the neuraxis. Several areas had a higher concentration of cells that express connexin43, such as layer IA of the piriform cortex, supraoptic and paraventricular nuclei of the hypothalamus, anterior cortical amygdaloid nucleus, the reticular part of the substantia nigra, lateral habenula, mesencephalic trigeminal nucleus. Purkinje cell layer of the cerebellum, facial nucleus, prepositus hypoglossal nucleus, and dorsal cochlear nucleus. The pattern of connexin43 hybridization and the morphology of connexin43 mRNA containing cells suggest that this gap junction forming protein is found predominantly in astrocytes. Connexin32 mRNA was detected in discrete cell groups of the gray matter that appeared to be neurons, including cells in layer 2 of the neocortex, layer II of the piriform cortex, pyramidal cell layer of the hippocampus, granule and polymorphic cell layers of the dentate gyrus, islands of Calleja, olfactory tubercle, lateral thalamic nuclei, lateral habenula, and Purkinje cell layer of the cerebellar cortex. A large population of cells in white matter tracts that were labelled with the connexin32 riboprobe appeared to be oligodendrocytes. These studies suggest that neurons and glial cells express connexin32 mRNA, but only astrocytes express connexin43 mRNA. Many of the areas in which connexin mRNAs were demonstrated have electrically coupled cells, morphologically distinct gap junction plaques, and/or have immunocytochemically identifiable connexin proteins. These results indicate that cells with mRNAs coding for intercellular channels have a widespread distribution in the mammalian CNS.

Animals↗

Manipulation of elementary charge in a silicon charge-coupled device.

The ultimate limit in the operation of an electronic device is the manipulation of a single charge. Such a limit has been achieved in single-electron tunnelling devices. However, these devices are based on multiple tunnel barriers and conductive islands, which are complex structures to fabricate. Here we demonstrate another type of device that can also manipulate elementary charge, but which is more suitable for large-scale integration. The device consists of two closely packed silicon wire-MOSFETs, which are commonly used building blocks of electronic circuits. We have developed a scheme to generate and store holes in the channels of either of these MOSFETs. Subsequently, holes can be transferred between the two MOSFETs at the level of an elementary charge, and their exact position can be monitored. This single-charge transfer device, which is operated at 25 K, is in effect a charge-coupled device. This is also the first realization of a silicon-based device that manipulates elementary charge.

Journal Article↗