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F Lai

Publications and source records attributed to F Lai.

At least 37 records · Page 2Linked to original sources
PubMed1997

Dramatic increase of the RNA editing for glutamate receptor subunits during terminal differentiation of clonal human neurons.

RNA editing plays an important role in determining physiological characteristics of certain glutamate-gated receptor (GluR) channels such as Ca2+ permeability and desensitization kinetics. In one case, the editing changes a gene-encoded glutamine (Q) to an arginine (R) codon located in the channel-forming domain of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor subunit GluR-B and also the kainate receptor subunits GluR5 and GluR6. Another case of RNA editing alters an arginine (R) to a glycine (G) codon at a position termed the "R/G" site of AMPA subunits GluR-B, C, and D. Double-stranded RNA-specific adenosine deaminases (DRADA) have been implicated as agents involved in the editing. By using a human teratocarcinoma cell line, NT2, we investigated the change of the RNA editing of GluR subunits in conjunction with the expression of two DRADA members, DRADA1 and DRADA2 genes, during neuronal differentiation. Whereas Q/R and R/G site RNA editing both become progressively activated in differentiating NT2 cells, the expression of the two DRADA genes can already be detected even in the undifferentiated NT2 cells. Development of the editing machinery appears to require, in addition to DRADA enzymes, a currently unidentified mechanism(s) that may become activated during neuronal differentiation.

Adenosine Deaminase↗
PubMed1997

Alzheimer-like visual deficits in Down syndrome.

Patients with Alzheimer disease (AD) show visual impairments in color discrimination (blue hues), stereoacuity, and contrast sensitivity. We asked whether the AD-type visual profile occurs in Down syndrome (DS) in light of the fact that AD neuropathology is present in DS by age 40. We tested 22 adults with DS and 18 adults with mental retardation of non-DS etiology (MR). DS subjects made more tritanomalous errors on the test of color vision than predicated by chance (p < 0.05), indicating a deficiency in the discrimination of short wavelengths (blue hues) but not more of other types of hue discrimination errors. DS subjects had higher stereoacuity thresholds than MR subjects (p < 0.01) and reduced contrast sensitivity across the frequency range (p < 0.01). Taken together, the results point to AD-like visual deficits in DS. Like classic AD, DS may be associated with pathological changes in the parastriate and peristriate visual cortex. DS performance was not correlated with age, suggesting that in individual subjects, the AD-like visual deficits may present prior to and independent of age-associated dementia.

Alzheimer Disease↗
PubMed1997

Diagnosis of dementia in individuals with intellectual disability.

The foremost impediment to progress in the understanding and treatment of dementia in adults with intellectual disability is the lack of standardized criteria and diagnostic procedures. Standardized criteria for the diagnosis of dementia in individuals with intellectual disability are proposed, and their application is discussed. In addition, procedures for determining whether or not criteria are met in individual cases are outlined. It is the intention of the authors, who were participants of an International Colloquium on Alzheimer Disease and Mental Retardation, that these criteria be appropriate for use by both clinicians and researchers. Their use will improve communication among clinicians and researchers, and will allow researchers to test hypotheses concerning discrepancies in findings among research groups (e.g. dementia prevalence ranges and age of onset).

Age of Onset↗
PubMed1997

Editing of glutamate receptor B subunit ion channel RNAs by four alternatively spliced DRADA2 double-stranded RNA adenosine deaminases.

Double-stranded (ds) RNA-specific adenosine deaminase converts adenosine residues into inosines in dsRNA and edits transcripts of certain cellular and viral genes such as glutamate receptor (GluR) subunits and hepatitis delta antigen. The first member of this type of deaminase, DRADA1, has been recently cloned based on the amino acid sequence information derived from biochemically purified proteins. Our search for DRADA1-like genes through expressed sequence tag databases led to the cloning of the second member of this class of enzyme, DRADA2, which has a high degree of sequence homology to DRADA1 yet exhibits a distinctive RNA editing site selectivity. There are four differentially spliced isoforms of human DRADA2. These different isoforms of recombinant DRADA2 proteins, including one which is a human homolog of the recently reported rat RED1, were analyzed in vitro for their GluR B subunit (GluR-B) RNA editing site selectivity. As originally reported for rat RED1, the DRADA2a and -2b isoforms edit GluR-B RNA efficiently at the so-called Q/R site, whereas DRADA1 barely edits this site. In contrast, the R/G site of GluR-B RNA was edited efficiently by the DRADA2a and -2b isoforms as well as DRADA1. Isoforms DRADA2c and -2d, which have a distinctive truncated shorter C-terminal structure, displayed weak adenosine-to-inosine conversion activity but no editing activity tested at three known sites of GluR-B RNA. The possible role of these DRADA2c and -2d isoforms in the regulatory mechanism of RNA editing is discussed.

Adenosine Deaminase↗
PubMed1997

[Detection of human cytomegalovirus infection in blood donor population by the polymerase chain reaction].

Samples of peripheral blood from 76 different donors were detected for the presence of human cytomegalovirus (HCMV) DNA by polymerase chain reaction (PCR). The results showed that HCMVs were present in 39 out of 76 blood donors which reached an infection positive rate of 51.3%. In addition, the sensitivity and specificity on PCR were tested, it also indicated that the sensitivity can reach the level of 5fg.microliter-1. The method is simple, rapid, highly sensitive and specificity. It can be considered as a diagnostic measure in clinical detection of HCMV.

Blood Donors↗
PubMed1996

Editing of the GLuR-B ion channel RNA in vitro by recombinant double-stranded RNA adenosine deaminase.

Double-stranded RNA (dsRNA)-specific adenosine deaminase (DRADA) has been implicated as an enzyme responsible for the editing of RNA transcripts encoding glutamate-gated ion channel subunits (GLuR) in brain. In one case, the editing alters the gene-encoded glutamine (Q) to an arginine (R) located within the channel-forming domain of the alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) receptor subunit GLuR-B. The result of editing at this site, called the 'Q/R' site, is a profound alteration of the Ca2+ permeability of the GLuR channel. Using recombinantly expressed DRADA proteins, we now demonstrate in vitro that DRADA is indeed involved in editing of the GLuR-B RNA. In addition to the formation of an RNA duplex structure involving exon and intron sequences, Q/R site-selective editing by DRADA also requires a cofactor protein(s) commonly present even in non-neuronal cells. The accuracy and efficiency of this RNA editing system appear to be determined by the quantitative balance between DRADA, cofactor and substrate GLuR-B RNA.

Adenosine Deaminase↗
PubMed1995

Mutagenic analysis of double-stranded RNA adenosine deaminase, a candidate enzyme for RNA editing of glutamate-gated ion channel transcripts.

Mutagenic analysis of the substrate binding and catalytic domains of double-stranded RNA (dsRNA) adenosine deaminase (DRADA) was carried out. This nuclear enzyme is likely to be involved in the RNA editing of glutamate-gated ion channels that are essential for fast excitatory neurotransmission in mammalian brain. The deletion of the first or the third of the three dsRNA binding motifs within the substrate binding domain dramatically decreases enzyme activity, whereas the second motif seems to be dispensable. The results indicate that the three motifs are not functionally equivalent in the catalytic action of DRADA. Mutation of the putative zinc-coordinating residues, His910, Cys966, and Cys1036, abolished the DRADA activity. Similarly, the Glu912 residue, predicted to be involved in the proton transfer functions of the enzyme, was found to be indispensable. Our results reinforce the previous proposal that the hydrolytic deamination mechanism of DRADA may be more similar to that of the cytidine deaminases than of adenosine deaminases.

Adenosine Deaminase↗
PubMed1995

Neuropathological changes in Down's syndrome hippocampal formation. Effect of age and apolipoprotein E genotype.

BACKGROUND: The neuropathological changes of Alzheimer's disease occur universally in individuals with Down's syndrome as they reach middle age and worsen with increasing age. Thus, evaluation of patients of various ages with Down's syndrome allows one to construct a life history of the development of neuropathological changes associated with Alzheimer's disease at various points in the disease process. METHODS: We have used semiquantitative scales and quantitative computerized image analysis techniques to analyze the characteristics of neurofibrillary tangle formation and A beta amyloid deposition in the hippocampal formation and inferior temporal gyrus in 36 individuals with Down's syndrome ranging in age from 4 to 73 years. RESULTS: Neurofibrillary tangles occur in a hierarchical distribution in a circumscribed set of neuronal fields, affecting the entorhinal cortex, area CA1/subiculum, then other hippocampal subfields. Although amyloid deposition occurs more evenly in a more widespread distribution, it also accumulates over the years 30 to 50. Surprisingly, examination of the patients available older than 50 years showed no trend toward continued increased deposition of amyloid. Within this group, however, individuals who had inherited the apolipoprotein E (Apo E) epsilon 4 genotype contained more than twice the amyloid burden of individuals who did not inherit the Apo E epsilon 4 genotype. COMMENT: This large series of cases confirms earlier observations that had suggested early vulnerability of entorhinal cortex and CA1/subiculum for neurofibrillary tangles and a more widespread but specific topography of A beta deposition. Moreover, it demonstrates quantitatively that the lesions increase to a certain level and then apparently reach a plateau. The level of amyloid deposition in Down's syndrome is higher than in sporadic Alzheimer's disease. Inheritance of the Apo E epsilon 4 genotype appears to be an additional (independent) risk factor for developing higher levels of amyloid accumulation.

Adolescent↗
PubMed1995

Scrub typhus pneumonitis with delayed resolution.

A 35-year-old lady was admitted to hospital with fever and dry cough. Chest radiograph showed bilateral basal infiltrate. Her Weil-Felix test was strongly positive (OX-K > 1:160) and her fever came down with intravenous tetracycline. There was no improvement in the lung shadow and spirometry showed a severe restrictive defect. Open lung biopsy confirmed the diagnosis of interstitial pneumonitis. CT of thorax 6 months after presentation showed partial resolution of the interstitial shadow.

Adult↗
PubMed1995

[Extrapulmonary tuberculosis. An unusual case].

Skeletal tuberculosis with extravertebral location is rare. We report a case of tuberculous osteomyelitis simultaneously affecting two right ribs and the left ulna. We emphasize the diagnostic problems (failure or delay in diagnosis) in extrapulmonary tuberculosis interesting uncommon sites and having relatively indolent presenting symptoms.

Aged↗
PubMed1994

Genomic organization and expressed sequences of the mouse extended H-2K region.

The mouse major histocompatibility complex (MHC) has long been of great interest to many biologists because of not only its critical role in the immune system, but also its association with at least three embryonic lethal genes. Here, we present an analysis of the mouse extended H-2K region using YAC technology. Six new expressed sequences were identified, demonstrating that the high gene density previously described continues. Restriction mapping of a YAC clone extending proximal of the MHC region defined a CpG-rich region located up to 320 kb away from H-2K. The absence of any CpG-rich region for a distance spanning approximately 200 kb near the YAC's proximal end suggests that the high gene density probably diminishes at a distance of 360 kb away from H-2K. The description of genomic organization of both H-2K and the extended H-2K region provides insight into the characteristics of this whole region with respect to gene diversity and density.

Animals↗
PubMed1994

Molecular analysis of mouse Rab11b: a new type of mammalian YPT/Rab protein.

Since the realization that the vesicular transport machinery is conserved from yeast to vertebrate neurons, much interest in the proteins involved has been generated. Here we describe a new type of mammalian YPT/Rab protein, named Rab11b, that is most abundantly expressed in brain, heart, and testis. It has all the hallmarks of a YPT3/Rab11 protein, but is more closely related to a ypt3-related gene isolated from fish (97% homology) than to the previously described four mammalian Rab11 genes (89% homology). The mammalian Rab11 genes discovered previously are, by contrast, 100% identical to each other. The genomic structure of a mammalian Rab11 protein is described for the first time. It is surprisingly asymmetrical, with 96% of the coding sequence contained in one-third of the transcription unit. Two copies of this gene are mapped to mouse chromosomes 1 and 17. The fine structural analysis of Rab11b and the protein sequence comparison provides a close look at all YPT3/Rab11-related genes cloned so far and clues to their possible relationship.

Amino Acid Sequence↗
PubMed1993

Chiral separation and detection enhancement of propranolol using automated pre-column derivatization.

In the liquid chromatographic analysis of pharmaceuticals, two challenges are often encountered: detectivity and chiral separations. Propranolol, a beta adrenergic blocker, is a pharmaceutical compound that faces both of these limitations. In this study, both limitations are overcome simultaneously using derivatization with (+)-1-(9-fluorenyl)ethyl chloroformate (FLEC), a highly fluorescent and chiral reagent. The derivatization is automated using an autosampler with an AutoMix microrobotic feature, which greatly contributes to the efficiency and reproducibility of the method when manipulating microliter volumes of sample and reagents. The method yields excellent separation of the diasteriomers, has a detection limit of 1 picomol, good reproducibility and linearity in the 50-400 pmol range (on column). In addition, this method is simple, requires no elevated temperature, no chiral stationary or mobile phases and can be easily automated.

Chromatography, High Pressure Liquid↗
PubMed1993

[Molecular genetic analysis on six cases of sex reversal syndrome].

Southern blot hybridization using Y-specific DNA probes and electrophoretic analysis of the SRY gene fragment amplified through polymerase chain reaction (PCR) have been performed in six cases of sex reversal syndrome (three 46, XX males and three 46, XY females). The result showed that the Y-specific DNA hybridization signal and SRY sequence were absent in one XX male and present in the remaining cases. This finding indicates that testis development of this XX male was initiated in the absence of the SRY gene.

Adult↗
PubMed1993

Matrix effects in the derivatization of amino acids with naphthalene dicarboxaldehyde, 9-fluorenylmethyl chloroformate and phenylisothiocyanate.

Pre-column derivatizations of amino acids often present two major challenges: 1) automation, due to the multi-step manipulations for pH control, reagent addition, mixing and extraction, and 2) effect of matrices in the sample such as salts, buffers and surfactants. Both issues have been addressed in a previous publication on derivatization methods using 9-fluorenylmethyl chloroformate (FMOC) and phenylisothiocyanate (PITC). In this paper, a third method of derivatization, which has recently been developed and published, was studied to address the same issues. The derivatization reagent, naphthalene-2,3-dicarboxaldehyde (NDA), is a modification from o-phthalaldehyde (OPA) and yields more stable derivatives with high fluorescence efficiencies. An autosampler was programmed to mix amino acid samples with cyanide and NDA reagent, allow a programmed reaction time and finally inject onto the HPLC. To study sample matrix effects, amino acid samples were spiked with various concentrations of Tris-HCl, phenol, citrate, sulfosalicylic acid, sodium chloride and sodium dodecyl sulfate. The recoveries of amino acids in varied sample matrices were compared to pure amino acid standards. The matrix effects using the NDA method were similar to those using the FMOC method. Comparisons of all three methods (NDA, FMOC and PITC) are discussed and tabulated.

Amino Acids↗
PubMed1992

Enhancement of detection sensitivity and cleanup selectivity for tobramycin through pre-column derivatization.

Reversed-phase high-performance liquid chromatography, UV detection and reversed-phase solid-phase extraction (SPE) as analytical methods for pharmaceutical compounds face a challenge when the compound is rather polar and lack UV absorptivity. A good example is tobramycin. To overcome these problems, a method has been developed using pre-column derivatization of tobramycin with o-phthalaldehyde and automated with an autosampler with microrobotic routines. The detection enhancement of the derivatives was achieved by using fluorescence detection which was forty times more sensitive than using UV detection. Recovery studies of standards and spiked serum samples show that pre-SPE derivatization significantly enhances the recoveries (by at least a factor of 3) and the quality of cleanup over post-SPE derivatization.

Chromatography, High Pressure Liquid↗
PubMed1992

New molecular markers for the distal end of the t-complex and their relationships to mutations affecting mouse development.

Many mutations affecting mouse development have been mapped to the t-complex of mouse chromosome 17. We have obtained 17 cosmid clones as molecular markers for this region by screening a hamster-mouse chromosome 17 and 18 cell hybrid cosmid library with mouse-specific repetitive elements and mapping positive clones via t-haplotype vs. C3H restriction fragment length polymorphism (RFLP) analysis. Twelve of the clones mapping distal to Leh66B in t-haplotypes are described here. Using standard RFLP analysis or simple sequence length polymorphism between t-haplotypes, exceptional partial t-haplotypes and nested sets of inter-t-haplotype recombinants, five cosmids have been mapped in or around In(17)3 and seven in the most distal inversion In17(4). More precise mapping of four of the cosmids from In(17)4 shows that they will be useful in the molecular identification of some of the recessive lethals mapped to the t-complex: two cosmids map between H-2K and Crya-1, setting a distal limit in t-haplotypes for the position of the tw5 lethal, one is inseparable from the tw12 lethal, and one maps distal to tf near the t0(t6) lethal and cld.

Animals↗