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PubMedDate not supplied

Quantitative ultrastructural analysis of perifibrous lipid and its association with elastin in nonatherosclerotic human aorta.

Nonatherosclerotic areas in human arteries display an age-related accumulation of cholesteryl ester in the form of small, perifibrous lipid droplets in the deeper intimal layers. We treated human aortic specimens with an osmium-thiocarbohydrazide-osmium sequence en bloc after glutaraldehyde fixation in order to provide electron dense staining of neutral lipid for ultrastructural study. Neutral lipid was quantified in terms of area fractions on thin sections. Extracellular lipid, primarily in the form of small (less than 300 nm) droplets, accounted for 91% of the lipid found in the deep intimal region. Seventy-four percent of extracellular lipid appeared in droplets or aggregates that were demonstrated as adjacent to or within elastic fibers in the plane of section. The fraction of lipid adjacent to elastin in three dimensions is likely to be considerably higher than 74%. The results support the concept that an interaction between elastin or its associated components and lipids or lipoproteins may be important in extracellular lipid deposition in human arteries.

Adolescent↗
PubMed2003

Subarachnoid hemorrhage without evident cause on initial angiography studies: diagnostic yield of subsequent angiography and other neuroimaging tests.

OBJECT: The aim of this study was to assess the diagnostic yield of imaging tests performed in patients in whom the cause of subarachnoid hemorrhage (SAH) had not been demonstrated on initial angiography. METHODS: By reviewing medical records of 806 patients with SAH who had been admitted during a 6.5-year period, the authors identified 86 in whom initial transfemoral catheter angiography failed to reveal the cause of SAH. Clinical and radiological data were analyzed to determine the diagnostic yield of subsequent catheter angiography, computerized tomography (CT) angiography, magnetic resonance (MR) angiography, and MR imaging of the brain and spine for various subtypes of SAH (bleeding not visualized on CT studies [CT-negative SAH], perimesencephalic SAH, and nonperimesencephalic SAH). Of 41 patients with nonperimesencephalic SAH, 36, 32, and 21 underwent repeated catheter angiography, CT angiography, and MR angiography, respectively; brain MR imaging was performed in 23 patients (18 with Gd and 15 with susceptibility contrast sequences), and spine MR imaging in 17. Of 36 patients with perimesencephalic SAH, 31, 23, and 17 underwent repeated catheter angiography, CT angiography, and MR angiography, respectively; brain MR imaging was performed in 18 patients (17 with Gd and 11 with susceptibility contrast sequences), and spine MR imaging in 14. Of nine patients with SAH not visualized on CT scanning, three, one, and six underwent repeated catheter angiography, CT angiography, and MR angiography, respectively; brain MR imaging was performed in eight patients (five with Gd and three with susceptibility contrast sequences), and spine MR imaging in seven. The cause of SAH could be determined in only four patients, all with nonperimesencephalic SAH. The only test that yielded a diagnosis was catheter angiography (three aneurysms on the second and one on the third angiography, all surgically secured). Diffusion-weighted MR imaging demonstrated small, deep infarcts in five patients. CONCLUSIONS: Repeated catheter angiography remains the most sensitive test to determine the cause of SAH that is not demonstrated on initial angiography, particularly in the subtype of nonperimesencephalic SAH. Newer, noninvasive imaging techniques provide little diagnostic yield.

Adult↗
PubMedDate not supplied

[Assessment with magnetic resonance of laryngeal and oropharyngeal movements during phonation].

The latest MR units are provided with the Turbo-Field-Echo technique which permits gradient-echo imaging with very short TE and TR and is optimized to yield the highest possible image quality within a very short acquisition time--i.e., less than 5 seconds. This dynamic study was aimed at depicting normal laryngeal and oropharyngeal movements during maximal inspiration and the prolonged uttering of vowels, in both a normal and a loud voice. We examined 10 healthy volunteers (7 men and 3 women) with an 0.5-T superconductive unit (Gyroscan T5 III, Philips Medical System). We used Turbo-Field-Echo sequences with the following acquisition parameters: TR 12 ms, TE 6 ms, flip angle 30 degrees, 4 acquisitions, acquisition time: 5 seconds. A single coronal scan was acquired at the larynx, while a midsagittal scan and 2 coronal scans were acquired at the oropharynx. The volunteers were asked to breathe in long and deep, to prolong the emission of the vowel [i] during laryngeal studies and to prolong the emission of the fundamental vowels [a], [i] and [u] in a normal (50 db) and a loud (70 db) voice during oropharyngeal studies. The movements of true and false vocal cords were clearly depicted in all the volunteers, and the activity of the different anatomical structures of the oropharyngeal cavity (lips, tongue, hard and soft palate, pharynx and epiglottis) was also demonstrated. During vowel production in a loud voice, the vocal tract was enlarged at the oral cavity for the vowels [a] and [u] and at the pharynx for the vowel [i]. To conclude, fast MRI with midsagittal scans is the best imaging modality to study different vocal tract patterns during speech and can thus replace midsagittal radiography and xeroradiography in the study of vowel production.

Adult↗
PubMed1989

Elective deep hypothermia with total circulatory arrest: changes in plasma creatine kinase BB, blood glucose, and clinical variables.

Serial measurements of a marker of brain ischemia, creatine kinase isoenzyme BB, were performed in arterial and internal jugular venous blood from 20 infants younger than 1 year of age before and during the first 20 hours after deep hypothermic total circulatory arrest procedures. A two-site monoclonal method was used, and the results were analyzed in relation to age, size, type of cardiac lesion, hemoglobin level, blood glucose level, pH, and duration of the total circulatory arrest. The creatine kinase BB concentrations increased after the arrest, more so in venous than in arterial blood, from 3.2 +/- 0.5 ng/ml to 17.5 +/- 4.5 in arterial blood and from 3.5 +/- 0.5 ng/ml to 18.1 +/- 5.8 in venous blood. Arterial-venous concentration differences correlated with venous concentrations (r = 0.92, p less than 0.01). The duration of the arrest correlated with creatine kinase BB concentrations during reperfusion with correlation coefficients between 0.50 to 0.90 depending of what sequence of the 20-hour sampling period was analyzed. The best correlation was obtained during the first 4 hours of reperfusion. Age, size of the child, and preoperative cyanosis correlated with postoperative creatine kinase BB but were less important than the arrest time, blood glucose level (r = 0.62, p less than 0.01), pH (r = -0.78, p less than 0.01), and hemoglobin level (r = 0.76, p less than 0.01) during reperfusion. It is suggested that a different control of blood glucose level and pH during reperfusion may be of importance to reduce biochemical signs of cerebral dysfunction after deep hypothermic total circulatory arrest procedures.

Blood Glucose↗
PubMed2006

A comparison of stable-isotope probing of DNA and phospholipid fatty acids to study prokaryotic functional diversity in sulfate-reducing marine sediment enrichment slurries.

Marine sediment slurries enriched for anaerobic, sulfate-reducing prokaryotic communities utilizing glucose and acetate were used to provide the first comparison between stable-isotope probing (SIP) of phospholipid fatty acids (PLFA) and DNA (16S rRNA and dsrA genes) biomarkers. Different 13C-labelled substrates (glucose, acetate and pyruvate) at low concentrations (100 microM) were used over a 7-day incubation to follow and identify carbon flow into different members of the community. Limited changes in total PLFA and bacterial 16S rRNA gene DGGE profiles over 7 days suggested the presence of a stable bacterial community. A broad range of PLFA were rapidly labelled (within 12 h) in the 13C-glucose slurry but this changed with time, suggesting the presence of an active glucose-utilizing population and later development of another population able to utilize glucose metabolites. The identity of the major glucose-utilizers was unclear as 13C-enriched PLFA were common (16:0, 16:1, 18:1omega7, highest incorporation) and there was little difference between 12C- and 13C-DNA 16S rRNA gene denaturing gradient gel electrophoresis (DGGE) profiles. Seemingly glucose, a readily utilizable substrate, resulted in widespread incorporation consistent with the higher extent of 13C-incorporation (approximately 10 times) into PLFA compared with 13C-acetate or 13C-pyruvate. 13C-PLFA in the 13C-acetate and 13C-pyruvate slurries were similar to each other and to those that developed in the 13C-glucose slurry after 4 days. These were more diagnostic, with branched odd-chain fatty acids (i15:0, a15:0 and 15:1omega6) possibly indicating the presence of Desulfococcus or Desulfosarcina sulfate-reducing bacteria (SRB) and sequences related to these SRB were in the 13C-acetate-DNA dsrA gene library. The 13C-acetate-DNA 16S rRNA gene library also contained sequences closely related to SRB, but these were the acetate-utilizing Desulfobacter sp., as well as a broad range of uncultured Bacteria. In contrast, analysis of DGGE bands from 13C-DNA demonstrated that the candidate division JS1 and Firmicutes were actively assimilating 13C-acetate. Denaturing gradient gel electrophoresis also confirmed the presence of JS1 in the 13C-DNA from the 13C-glucose slurry. These results demonstrate that JS1, originally found in deep subsurface sediments, is more widely distributed in marine sediments and provides the first indication of its metabolism; incorporation of acetate and glucose (or glucose metabolites) under anaerobic, sulfate-reducing conditions. Here we demonstrate that PLFA- and DNA-SIP can be used together in a sedimentary system, with low concentrations of 13C-substrate and overlapping incubation times (up to 7 days) to provide complementary, although not identical, information on carbon flow and the identity of active members of an anaerobic prokaryotic community.

DNA Fingerprinting↗
PubMed2005

Papillomavirus subtypes are natural and old taxa: phylogeny of human papillomavirus types 44 and 55 and 68a and -b.

A human papillomavirus (HPV) type is defined as an HPV isolate whose L1 gene sequence is at least 10% different from that of any other type, while a subtype is 2 to 10% different from any HPV type. In order to analyze the phylogeny behind the subtype definition, we compared 49 isolates of HPV type 44 (HPV-44) and its subtype HPV-55, previously misclassified as a separate type, and 41 isolates of the subtype pair HPV-68a and -b, sampled from cohorts in four continents. The subtypes of each pair are separated by deep dichotomic branching, and three of the four subtypes have evolved large phylogenetic clusters of genomic variants forming a "star" phylogeny, with some branches specific for ethnically defined cohorts. We conclude that subtypes of HPV types are natural and old taxa, equivalent to types, which either diverged more recently than types or evolved more slowly.

Molecular Sequence Data↗
PubMed2005

An empirical assessment of long-branch attraction artefacts in deep eukaryotic phylogenomics.

In the context of exponential growing molecular databases, it becomes increasingly easy to assemble large multigene data sets for phylogenomic studies. The expected increase of resolution due to the reduction of the sampling (stochastic) error is becoming a reality. However, the impact of systematic biases will also become more apparent or even dominant. We have chosen to study the case of the long-branch attraction artefact (LBA) using real instead of simulated sequences. Two fast-evolving eukaryotic lineages, whose evolutionary positions are well established, microsporidia and the nucleomorph of cryptophytes, were chosen as model species. A large data set was assembled (44 species, 133 genes, and 24,294 amino acid positions) and the resulting rooted eukaryotic phylogeny (using a distant archaeal outgroup) is positively misled by an LBA artefact despite the use of a maximum likelihood-based tree reconstruction method with a complex model of sequence evolution. When the fastest evolving proteins from the fast lineages are progressively removed (up to 90%), the bootstrap support for the apparently artefactual basal placement decreases to virtually 0%, and conversely only the expected placement, among all the possible locations of the fast-evolving species, receives increasing support that eventually converges to 100%. The percentage of removal of the fastest evolving proteins constitutes a reliable estimate of the sensitivity of phylogenetic inference to LBA. This protocol confirms that both a rich species sampling (especially the presence of a species that is closely related to the fast-evolving lineage) and a probabilistic method with a complex model are important to overcome the LBA artefact. Finally, we observed that phylogenetic inference methods perform strikingly better with simulated as opposed to real data, and suggest that testing the reliability of phylogenetic inference methods with simulated data leads to overconfidence in their performance. Although phylogenomic studies can be affected by systematic biases, the possibility of discarding a large amount of data containing most of the nonphylogenetic signal allows recovering a phylogeny that is less affected by systematic biases, while maintaining a high statistical support.

Animals↗
PubMed2003

Subthalamic nucleus deep brain stimulation for Parkinson's disease: magnetic resonance imaging targeting using visible anatomical landmarks.

BACKGROUND: The aim of the present study was to validate a magnetic resonance imaging (MRI) visual procedure to target the subthalamic nucleus (STN) based on surrounding anatomical landmarks. METHODS: 31 consecutive bilaterally implanted parkinsonian patients were included in this study. After identification of the anterior commissure (AC), posterior commissure (PC) and midcommissural point on a three-dimensional T1-weighted sequence, inversion recovery (IR) T2-weighted coronal slices were performed orthogonal to the AC-PC line. On the slice showing the anterior pole of the red nucleus (RN), the target was placed in the inferolateral portion of the subthalamic zone, limited superiorly by the thalamus, laterally by the internal capsule, inferiorly by the substantia nigra and medially by the midline. The distribution of the targets was analyzed in the AC-PC referential. RESULTS: The mean target coordinates were as follows: anteroposterior (AP) = -2.54 mm (+/-1.37 mm), lateral (LAT) = 12.03 mm (+/-0.91 mm) and vertical (VERT) = -6.10 mm (+/-1.52 mm) for the right side, and AP = -2.65 mm (+/-1.36 mm), LAT = -11.97 mm (+/-1.30 mm) and VERT = -5.89 mm (+/-1.52 mm) for the left side. They projected in the inferior portion of the STN on the Schaltenbrand and Wahren atlas [Stuttgart, Thieme, 1977]. CONCLUSION: Identification of the anterior pole of the RN and the subthalamic zone on coronal IR T2-weighted MRI performed orthogonal to the AC-PC line provides a precise visual procedure to target the STN.

Electric Stimulation Therapy↗
PubMed1994

Topology of an amphiphilic mitochondrial signal sequence in the membrane-inserted state: a spin labeling study.

To investigate the interaction of the presequence of the precursor of yeast cytochrome C oxidase subunit IV (COX IV) with phospholipid membranes, a series of single- and double-cysteine-substituted peptide variants derived from the 25-residue NH2-terminal presequence has been synthesized and modified with nitroxide spin labels. The immersion depth, orientation, and secondary structure of the peptide in the POPC bilayer containing 10 mol % POPG were determined using electron paramagnetic resonance (EPR) spectroscopy. EPR saturation analysis of singly labeled variants reveals that the nitroxides attached to the NH2-terminal region of the peptide insert into the acyl chain region of the bilayer, approximately 13 A deep from the membrane surface. EPR line shape analysis of doubly labeled variants indicates that the peptide predominantly exists as an extended conformation, with little secondary structure. The experimental results, together with the energetic consideration of peptide-bilayer interactions, suggest that the presequence is located near the interface between the head group region and the acyl chain region, such that the hydrophobic side chains are solvated by the acyl chains and the charged side chains extended toward the polar environment at the bilayer surface.

Amino Acid Sequence↗
PubMed2006

A chloroplast genealogy of myrmecophytic Macaranga species (Euphorbiaceae) in Southeast Asia reveals hybridization, vicariance and long-distance dispersals.

Macaranga (Euphorbiaceae) includes about 280 species with a palaeotropic distribution. The genus not only comprises some of the most prominent pioneer tree species in Southeast Asian lowland dipterocarp forests, it also exhibits a substantial radiation of ant-plants (myrmecophytes). Obligate ant-plant mutualisms are formed by about 30 Macaranga species and 13 ant species of the genera Crematogaster or Camponotus. To improve our understanding of the co-evolution of the ants and their host plants, we aim at reconstructing comparative organellar phylogeographies of both partners across their distributional range. Preliminary evidence indicated that chloroplast DNA introgression among closely related Macaranga species might occur. We therefore constructed a comprehensive chloroplast genealogy based on DNA sequence data from the noncoding ccmp2, ccmp6, and atpB-rbcL regions for 144 individuals from 41 Macaranga species, covering all major evolutionary lineages within the three sections that contain myrmecophytes. A total of 88 chloroplast haplotypes were identified, and grouped into a statistical parsimony network that clearly distinguished sections and well-defined subsectional groups. Within these groups, the arrangement of haplotypes followed geographical rather than taxonomical criteria. Thus, up to six chloroplast haplotypes were found within single species, and up to seven species shared a single haplotype. The spatial distribution of the chloroplast types revealed several dispersals between the Malay Peninsula and Borneo, and a deep split between Sabah and the remainder of Borneo. Our large-scale chloroplast genealogy highlights the complex history of migration, hybridization, and speciation in the myrmecophytes of the genus Macaranga. It will serve as a guideline for adequate sampling and data interpretation in phylogeographic studies of individual Macaranga species and species groups.

Animals↗
PubMed2000

Vertebrate ancient-long opsin: a green-sensitive photoreceptive molecule present in zebrafish deep brain and retinal horizontal cells.

Nonretinal/nonpineal photosensitivity has been found in the brain of vertebrates, but the molecular basis for such a "deep brain" photoreception system remains unclear. We conducted an extensive search for brain opsin cDNAs of the zebrafish (Danio rerio), a useful animal model for genetic studies, and we have isolated a partial cDNA clone encoding an ortholog of vertebrate ancient (VA) opsin, the function of which is unknown. Subsequent characterization revealed the occurrence of two kinds of mRNAs encoding putative splicing variants, VA and VA-Long (VAL) opsin, the latter of which is a novel variant of the former. Both opsins shared a common core sequence in the membrane-spanning domains, but VAL-opsin had a C-terminal tail much longer than that of VA-opsin. Functional reconstitution experiments on the recombinant proteins showed that VAL-opsin with bound 11-cis-retinal is a green-sensitive pigment (lambdamax approximately 500 nm), whereas VA-opsin exhibited no photosensitivity even in the presence of 11-cis-retinal. Immunoreactivity specific to this functionally active VAL-opsin was localized at a limited number of cells surrounding the diencephalic ventricle of central thalamus, and these cells were distributed over approximately 200 micrometer along the rostrocaudal axis. Taken together with the previous study on the locus of the teleost brain photosensitivity (von Frisch K, 1911), it is strongly suggested that the VAL-positive cells in the zebrafish brain represent the deep brain photoreceptors. The VAL-specific immunoreactivity was also detected in a subset of non-GABAergic horizontal cells in the zebrafish retina. The existence of VAL-opsin, a new member of the rhodopsin superfamily, in these tissues may indicate its multiple roles in visual and nonvisual photosensory physiology.

Animals↗
PubMed1995

Structure and expression of the bovine oxytocin receptor gene.

The gene for the bovine oxytocin receptor has been sequenced using a combination of clones derived from a bovine endometrial cDNA library from estrus and a bovine genomic DNA library, with confirmation of structure using reverse transcription PCR programmed by term myometrial RNA. The receptor belongs to the seven transmembrane domain family and predicts a protein of 391 amino acids. A comparison of the genomic sequence with the cDNA structure, as well as reverse transcription polymerase chain reaction (RT-PCR) analysis, shows there are two introns, one in the 5'noncoding region that appears to be differentially spliced in the bovine uterus and a conserved intron within the open reading frame between the regions encoding the transmembrane domains VI and VII. Northern blot analysis indicated three major transcripts in myometrium and endometrium in vivo at approximately 6.5 kb, 3.5 kb, and 2.0 kb. In situ hybridization analysis of uterine tissue at term showed highest mRNA concentrations in the endometrial epithelium, particularly in the deep glands, a pattern confirmed also at the immunohistochemical level by monoclonal antibodies raised against a human amino-terminal peptide. Further confirmation of the identity of the receptor was obtained by transient transfection of a reconstituted receptor construct into COS-7 cells. The expressed receptor was shown to have identical pharmacological properties in respect to various oxytocin analogs to the natural bovine endometrial receptor.

Amino Acid Sequence↗
PubMed2001

The sclerophytin A adventure.

The structural designation A originally made by Sharma and Alam to sclerophytin A was considered to be ambiguous and so notably strained relative to B that the latter was targeted for de novo synthesis (Scheme 1). Our two successful routes began with (5S)-(d-menthyloxy)-2(5H)-furanone and involved the application of cycloaddition, Claisen ring expansion, transannular oxymercuration, and 1,2-carbonyl transposition tactics to arrive at B. It was immediately apparent from polarity considerations and spectroscopic data that the antileukemic marine metabolite in question was in need of more deep-seated structural revision. Following close re-examination of an acquired authentic sample by advanced NMR techniques, the strong inference was made that sclerophytin A actually lacked a second oxygen bridge and was in reality the triol C. This conclusion was unequivocally confirmed by diverting an advanced intermediate generated earlier into a short sequence beginning with regiocontrolled dihydroxylation and terminating with configurational inversion at the secondary carbinol center. The status of other members of this series is also presented.

Animals↗
PubMed1987

Phylogenetic diversity and position of the genus Campylobacter.

RNA sequence analysis has been used to examine the phylogenetic position and structure of the genus Campylobacter. A complete 5S rRNA sequence was determined for two strains of Campylobacter jejuni and extensive partial sequences of the 16S rRNA were obtained for several strains of C. jejuni and Wolinella succinogenes. In addition limited partial sequence data were obtained from the 16S rRNAs of isolates of C. coli, C. laridis, C. fetus, C. fecalis, and C. pyloridis. It was found that W. succinogenes is specifically related to, but not included, in the genus Campylobacter as presently constituted. Within the genus significant diversity was noted. C. jejuni, C. coli and C. laridis are very closely related but the other species are distinctly different from one another. C. pyloridis is without question the most divergent of the Campylobacter isolates examined here and is sufficiently distinct to warrant inclusion in a separate genus. In terms of overall position in bacterial phylogeny, the Campylobacter/Wolinella cluster represents a deep branching most probably located within an expanded version of the Division containing the purple photosynthetic bacteria and their relatives. The Campylobacter/Wolinella cluster is not specifically includable in either the alpha, beta or gamma subdivisions of the purple bacteria.

Base Sequence↗
PubMed2003

Insights into metabolism obtained from microarray analysis.

The regulation of plant metabolic processes in response to environmental and developmental signals is a complex interaction between optimization of enzyme activity and transcriptional regulation of gene expression. Through painstaking efforts over more than 50 years, many metabolic pathways in plants have been characterized and their regulation investigated, often in detail. Widely available cDNA and oligonucleotide arrays can now be used to analyze the expression profiles of a large number of genes in parallel. Coupling these expression profiles to detailed analyzes of metabolite changes will allow deep insight into the cellular mechanisms of metabolic adaptation to a wide variety of growth conditions.

Adaptation, Physiological↗
PubMed2003

Lupus-like panniculitis in a patient with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED).

Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare autosomal recessive disorder, characterised by a loss of self-tolerance to endocrine tissues, chronic candidiasis and ectodermal disorders. APECED is associated with mutations of a single gene, designated autoimmune regulator (AIRE). We describe a 31-year-old APECED patient with non-traumatic, cutaneous ulcers on both forearms with features of a lupus-like panniculitis. On admission to the ICU in September 2001, the patient suffered from a ketoacidotic, hyperglycemic coma and adrenal crisis due to an Enterobacter-cloacae sepsis, originating from multiple, necrotising deep cutaneous ulcers. These ulcers spontaneously developed on both forearms, some of which were just emerging, full blown or healing with scars. Histological examination showed signs of a scarring panniculitis and vasculitis. Immunohistochemistry and direct immunofluorescence with characterisation of immunoglobulin and complement-factor binding pattern revealed features of a lupus-like panniculitis. Sequence analysis of all 14 exons of the AIRE gene revealed a R257 X mutation in exon 6 resulting in a nonsense mutation at codon 257 confirming the diagnosis of APECED. Oral treatment with 60 mg/day corticosteroids for two weeks led to complete resolution of all ulcers. In conclusion, mutations in the AIRE gene may provide the genetic background against which additional factors can initiate an autoimmune process. Here, autoimmune panniculitis appears to be an associated feature of the APECED syndrome. Our findings support the use of immunosuppressive therapy for autoimmune disease components of the APECED syndrome.

Adult↗
PubMed2003

Biosynthesis of isoprenoids: crystal structure of 4-diphosphocytidyl-2C-methyl-D-erythritol kinase.

4-Diphosphocytidyl-2C-methyl-d-erythritol kinase, an essential enzyme in the nonmevalonate pathway of isopentenyl diphosphate and dimethylallyl diphosphate biosynthesis, catalyzes the single ATP-dependent phosphorylation stage affording 4-diphosphocytidyl-2C-methyl-d-erythritol-2-phosphate. The 2-A resolution crystal structure of the Escherichia coli enzyme in a ternary complex with substrate and a nonhydrolyzable ATP analogue reveals the molecular determinants of specificity and catalysis. The enzyme subunit displays the alpha/beta fold characteristic of the galactose kinase/homoserine kinase/mevalonate kinase/phosphomevalonate kinase superfamily, arranged into cofactor and substrate-binding domains with the catalytic center positioned in a deep cleft between domains. Comparisons with related members of this superfamily indicate that the core regions of each domain are conserved, whereas there are significant differences in the substrate-binding pockets. The nonmevalonate pathway is essential in many microbial pathogens and distinct from the mevalonate pathway used by mammals. The high degree of sequence conservation of the enzyme across bacterial species suggests similarities in structure, specificity, and mechanism. Our model therefore provides an accurate template to facilitate the structure-based design of broad-spectrum antimicrobial agents.

Adenosine Triphosphate↗
PubMed2001

A deep intronic mutation in CDKN2A is associated with disease in a subset of melanoma pedigrees.

Germline mutations of CDKN2A at 9p21 have been shown to predispose to disease in melanoma pedigrees worldwide. However, there remains a significant proportion of melanoma pedigrees with evidence of linkage to 9p21 in which mutations in CDKN2A have not been detected. Investigation of other potential tumour suppressor genes at 9p21 and the promotor of CDKN2A has been unable to explain genetic predisposition to melanoma in these pedigrees. Here we describe a mutation, IVS2-105 A/G, deep in intron 2 of CDKN2A, detected in six English melanoma pedigrees. The mutation creates a false GT splice donor site 105 bases 5' of exon 3 and has been demonstrated to result in aberrant splicing of the mRNA. This is the most common mutation identified in English families to date. The presence of this deep intronic mutation in a relatively large number of kindreds, indicates that it may account for a significant proportion of 9p21-linked melanoma pedigrees with no detectable mutations in the coding region of CDKN2A. In addition, the identification of one deep intronic mutation in CDKN2A indicates the possibility of the existence of other similar splicing mutations located elsewhere in the CDKN2A introns.

Alleles↗