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Biomedical subjects

S C Lo

Publications and source records attributed to S C Lo.

At least 55 records · Page 3Linked to original sources

Brain myo-inositol monophosphatase: activity of the single subunit in a dimeric enzyme.

Reversible dissociation of the dimeric structure of brain myo-inositol monophosphatase into subunits was attained by the addition of guanidine-HCl (4M). The molecular mass of the subunits (29 KDa) was determined by HPLC chromatography. Separation of the processes of refolding and association of the monomeric species was achieved by attaching the protein subunits to a rigid matrix (Affi-gel 15). The matrix-bound monomer is determined to be catalytically active, indicating that monomeric subunit of the phosphate is capable of conducting independent catalysis.

Animals↗

Identification of a putative infC-rpmI-rplT operon flanked by long inverted repeats in Mycoplasma fermentans (incognitus strain).

A specific 1542-bp DNA fragment was amplified from Mycoplasma fermentans (incognitus strain) using a unique 23-nucleotide (nt) synthetic deoxyribonucleotide (oligo) (5'-TCCAAAAAGTCCGGAATTTGGGG) as the primer pair in the polymerase chain reaction (PCR). The 23-nt sequence is part of the 29-bp terminal inverted repeat (IR) which forms the left potential stem-and-loop (s&l) structure of the previously identified M. fermentans insertion-sequence(IS)-like genetic element [Hu et al., Gene 93 (1990) 67-72]. The amplified DNA was cloned and sequenced. A pair of 27-bp IR containing the 23-nt synthetic oligo was identified at both termini. Between the IR, there are four potential open reading frames (ORFs) which are arranged adjacent to each other in the order, ORF-1, ORF-2, ORF-3 and ORF-4, with parts of ORF-1 and ORF-2 overlapping. The deduced amino acid (aa) sequences of ORF-2, ORF-3 and ORF-4 are 34 to 60% identical to the translation initiation factor IF3 (encoded by the infC gene), ribosomal proteins L35 (rpmI gene) and L20 (rplT gene) of Escherichia coli and Bacillus stearothermophilus, respectively. In bacteria, the infC-rpmI-rplT genes are organized to function as an operon. There are multiple sites with promoter-like sequences identified upstream from the putative infC gene in the mycoplasma closely resembling the gene arrangement in the bacterial operon. All three genes of ORF-2, ORF-3 and ORF-4 are preceded individually by a strong appropriately spaced (7 and 10 bp) putative Shine-Dalgarno sequence (5'-AAGGA).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Reversible unfolding of myo-inositol monophosphatase.

myo-Inositol monophosphatase isolated from pig brain is a very stable dimeric protein characterized by a rotational correlation time of 30 ns. The unfolding and dissociation of the dimeric enzyme (58 kDa) by guanidine hydrochloride have been investigated at equilibrium. The overall process was reversible as judged from the complete recovery of catalytic activity after dilution of guanidine hydrochloride-treated samples. Unfolding of myo-inositol monophosphatase was monitored by circular dichroism, fluorescence, and steady state emission anisotropy. A folded, monomeric form of the monophosphatase was not detected by the method of denaturant gel filtration. Uncoupled dissociation and unfolding of the oligomeric enzyme could not be demonstrated. The circular dichroism and emission anisotropy results are consistent with a model in which the dimeric protein unfolds in a single cooperative transition from folded dimer to two unfolded monomers.

Animals↗

Automated drug dissolution monitor that uses a UV-visible diode array spectrophotometer.

A method for performing multicomponent analysis in drug dissolution testing without chromatographic separation is presented. Aliquots from dissolution vessels are automatically transferred to a UV-visible diode array spectrophotometer, spectra are measured, and the aliquots are returned to the testing vessels. A full-spectrum calibration method based on principal-component regression is used to simultaneously determine the concentrations of active ingredients and to account for interferences due to excipients in a tablet formulation. The system was evaluated with two commercial pharmaceutical formularies; the first contained pseudoephedrine hydrochloride and chlorpheniramine maleate, whereas the second was a mixture of phenylpropanolamine hydrochloride and chlorpheniramine maleate. The selections of standard mixtures for calibration and validation were based on a factorial design.

Calibration↗

Automatic lung nodule detection using profile matching and back-propagation neural network techniques.

The potential advantages of using digital techniques instead of film-based radiography have been discussed extensively for the past 10 years. A major future application of digital techniques is computer-assisted diagnosis: the use of computer techniques to assist the radiologist in the diagnostic process. One aspect of this assistance is computer-assisted detection. The detection of small lung nodule has been recognized as a clinically difficult task for many years. Most of the literature has indicated that the rate for finding lung nodules (size range from 3 mm to 15 mm) is only approximately 65%, in those cases in which the undetected nodules could be found retrospectively. In recent published research, image processing techniques, such as thresholding and morphological analysis, have been used to enhance true-positive detection. However, these methods still produce many false-positive detections. We have been investigating the use of neural networks to distinguish true-positives nodule detections among those areas of interest that are generated from a signal enhanced image. The initial results show that the trained neural networks program can increase true-positive detections and moderately reduce the number of false-positive detections. The program reported here can perform three modes of lung nodule detection: thresholding, profile matching analysis, and neural network. This program is fully automatic and has been implemented in a DEC 5000/200 (Digital Equipment Corp, Maynard, MA) workstation. The total processing time for all three methods is less than 35 seconds. In this report, key image processing techniques and neural network for the lung nodule detection are described and the results of this initial study are reported.

Algorithms↗

Vector quantization distortion of medical ultrasound features.

Pruned-tree structured vectored quantization (PTSVQ) was applied to the lower five gray scale remapped bits of normal and fatty ultrasound liver images. The upper bits were compressed reversibly. This combination of techniques is termed PTSVQ with splitting. The effect of the compression on the difference in texture between normal and fatty liver images was studied at different compression rates and distortions. The changes in texture were measured by changes in the principal components of the covariance matrix of image vectors. The vectors were the same size as those used in the compression technique. There were clear differences in the components of normal and fatty liver images. These differences were largely removed by the PTSVQ with splitting technique even at average single pixel distortions several times smaller than the image noise. These results suggest that the effect of compression on second order statistics should be measured when evaluating algorithms in addition to the first order average distortion.

Fatty Liver↗

Acquired immunodeficiency syndrome-like illness associated with systemic Mycoplasma fermentans infection in a human immunodeficiency virus-negative homosexual man.

A 35-year-old homosexual man developed a composite nodal Kaposi's sarcoma and peripheral T-cell lymphoma that were associated with a peripheral blood CD4-positive lymphocyte count of only 43/mm3. The patient subsequently developed Pneumocystis carinii pneumonitis and eventually died due to disseminated Cryptococcus neoformans. Numerous premortem tests for the presence of human immunodeficiency virus (HIV) types 1 and 2 were negative by the enzyme-linked immunosorbent assay, Western blot, viral isolation, and polymerase chain reaction techniques. Postmortem evaluations for HIV-1, HIV-2, human T-cell lymphotropic virus (HTLV)-I, and HTLV-II also were negative by polymerase chain reaction, immunofluorescence assays, and viral isolation. A systemic infection by Mycoplasma fermentans, however, was documented by immunohistochemistry and polymerase chain reaction in premortem and postmortem tissues. This recently recognized human pathogen has produced systemic infections in patients with the acquired immunodeficiency syndrome (AIDS) and in previously healthy non-AIDS patients who characteristically have a fulminant flu-like illness. Additionally, M fermentans has enhanced the cytopathic effect of HIV in in vitro studies and has produced fatal wasting illnesses with terminal lymphopenia in inoculated adult silvered leaf monkeys. This report is the first description of an association between M fermentans infection and an AIDS-like illness in an HIV-negative individual. The etiology of the severe immunosuppression in this patient and the associated role of M fermentans remain to be determined by further investigations.

Acquired Immunodeficiency Syndrome↗

Titers of antibody to Mycoplasma in sera of patients infected with human immunodeficiency virus.

One hundred sixty-seven sera collected from human immunodeficiency virus (HIV)-negative blood donors (n = 41), asymptomatic HIV-positive patients (n = 75), and patients with AIDS (n = 51) were compared in a metabolism inhibition serological test employing antigens to four distinct Mycoplasma species. The proportion of sera with antibody titers of > or = 1:32 to Mycoplasma genitalium was significantly higher for patients with AIDS than for HIV-negative blood donors. Serological relationships between Mycoplasma pneumoniae and M. genitalium as well as the possibility of increased susceptibility of AIDS patients to infection with these two mycoplasmas might account for this finding. Few sera showed antibody responses to Mycoplasma pirum or Mycoplasma fermentans.

AIDS-Related Opportunistic Infections↗

Adult respiratory distress syndrome with or without systemic disease associated with infections due to Mycoplasma fermentans.

An unusual but typical clinicopathological presentation was noted among several groups of previously healthy, human immunodeficiency virus-negative patients who had an apparent fulminant infectious disease but had no etiologic agent identified. The clinical courses were characterized by rapid progression and development of adult respiratory distress syndrome with or without systemic disease. Histopathological changes in the diseases tissues (other than lung) obtained by biopsy or at autopsy ranged from extensive necrosis with only minimal inflammatory reaction to prominent lymphohistiocytic infiltrate with focal areas of acute inflammation. This report focuses on pulmonary changes in three patients. The alveolar epithelial (type I) cells and type II pneumocytes are diffusely damaged. There is interstitial edema and thickened septa. An eosinophilic alveolar membrane may form, and the alveolar space may fill with foamy macrophages. Immunohistochemical studies identified Mycoplasma fermentans infection in the patients' lungs and liver. Mycoplasma-like particles could also be identified by electron microscopy. There is a previously unrecognized form of fulminant disease in humans that is associated with M. fermentans infections.

Adult↗

Fatal systemic infections of nonhuman primates by Mycoplasma fermentans (incognitus strain).

Four silvered leaf monkeys inoculated with Mycoplasma fermentans (incognitus strain) showed wasting syndromes and died in 7-9 months. Infected animals had a late and transient antibody response to mycoplasmal infection. Three monkeys revealed periodic mycoplasmal antigenemia. The one that had the most persistent antigenemia failed to mount a detectable antibody response and was the first to die of the infection. The control monkey was killed 8 months later, after the last of the infected animals had died, and revealed no evidence of seroconversion or antigenemia. Polymerase chain reaction, immunohistochemical, and electron microscopic studies identified systemic infections of M. fermentans in the infected animals. No other opportunistic infection or neoplastic disease was found. It is interesting to note the absence of an inflammatory reaction to the large number of mycoplasmas in the infected tissues. M. fermentans (incognitus strain) apparently suppressed normal inflammatory or immune responses, produced wasting syndromes, and caused a fatal systemic infection in these monkeys.

Animals↗

In vitro antibiotic susceptibility testing of different strains of Mycoplasma fermentans isolated from a variety of sources.

The in vitro susceptibilities to antibiotics of 24 strains of Mycoplasma fermentans (isolated from human immunodeficiency virus type 1-infected AIDS patients, non-AIDS patients with acute respiratory disease, and tissue culture) were determined. MICs for 90% of the strains tested (micrograms per milliliter) were obtained for chloramphenicol (1.25), ciprofloxacin (0.078), clindamycin (0.078), doxycycline (0.625), erythromycin (> 10), gentamicin (> 10), levofloxacin (0.078), lincomycin (0.156), streptomycin (> 10), and tetracycline (0.625).

Acquired Immunodeficiency Syndrome↗

A family of strain-variant surface lipoproteins of Mycoplasma fermentans.

The wall-less procaryote Mycoplasma fermentans is currently being examined as an agent potentially associated with human disease, including infectious processes affecting immunocompromised individuals. To delineate and understand the interactions of M. fermentans with its host, specific membrane surface components were characterized as markers for detecting the organism and for assessing heterogeneity in antigenic surface architecture within this mycoplasma species. Detergent phase fractionation of metabolically labeled organisms of type strain PG18 identified a family of prominent integral membrane proteins; several of these labeled with 35S-cysteine and 3H-palmitate, which are characteristics of procaryotic lipoproteins. Specific monoclonal and polyclonal antibodies raised to strain PG18 components further distinguished seven of these membrane proteins, which were localized on the organism's surface by monitoring their selective susceptibility during trypsin treatment of intact cells. With these antibodies, Western immunoblot profiles of surface membrane antigens expressed on strain PG18 were compared with those expressed on the recently identified Incognitus strain of M. fermentans, as well as with several other human and animal mycoplasma species. While the antibodies were specific for M. fermentans, marked differences were observed between the strains in the size of one surface lipoprotein and in the apparent levels of several antigens expressed in the cultured populations analyzed. Some monoclonal antibodies to strain PG18 and a previously described monoclonal antibody to strain Incognitus showed apparent selectivity for the strain used for immunization. Monoclonal antibodies developed here recognize stable epitopes defining a family of surface lipoproteins and provide critical tools to determine the basis of surface variation in this mycoplasma species and to assess the location and antigenic phenotypes of organisms in the human host.

Animals↗

Adhesion onto and invasion into mammalian cells by mycoplasma penetrans: a newly isolated mycoplasma from patients with AIDS.

The newly identified mycoplasma, Mycoplasma pentrans shows remarkable pathobiologic properties: it adheres to cell surfaces, deeply penetrates into the cell, strongly hemadsorbs human red blood cells, and cytadsorbs human CD4+ lymphocytes and monocytes. These in vitro biologic activities of mycoplasmas have been previously shown to be associated with pathogenic virulence in vivo. Both adhesion and invasion clearly involve the organism's unique tip-like structure. Invading mycoplasmas often have their tip-like structure deeply buried in the cytoplasm of infected mammalian cells. Extensive invasion of the mycoplasma into the cytoplasm may kill the cells. The same pathobiologic processes of adhesion and invasion using the specialized tip-like structure are found on the epithelium in the patient's urogenital tract infected by M. penetrans. Both in vitro and in vivo findings suggest a possible pathogenic role of this newly discovered human mycoplasma and call for careful evaluation of its role in human diseases.

Acquired Immunodeficiency Syndrome↗

Detection and isolation of Mycoplasma fermentans from urine of human immunodeficiency virus type 1-infected patients.

Mycoplasma fermentans (incognitus strain) has been linked with acquired immunodeficiency syndrome-associated nephropathy. Ten (23%) of 43 urine sediment specimens from 40 human immunodeficiency virus (HIV)-positive patients at different stages of acquired immunodeficiency syndrome disease tested positive in the polymerase chain reaction using a primer pair found in the insertion sequences specific to M fermentans. Mycoplasma fermentans was isolated from two HIV-positive patients' urine sediment specimens and on a repeated basis from one. All three culture-positive urine sediment specimens tested positive in the polymerase chain reaction. Fifty urine sediment specimens from age-matched HIV-negative healthy controls tested negative for M fermentans by polymerase chain reaction. Mycoplasma fermentans was not isolated from any of the control urine sediment specimens. Our results show a high prevalence of M fermentans in urine sediment specimens from HIV-positive patients but not from urine sediment specimens of HIV-negative healthy controls.

Acquired Immunodeficiency Syndrome↗

High frequency of antibodies to Mycoplasma penetrans in HIV-infected patients.

Mycoplasma penetrans, a novel mycoplasma isolated from HIV-1-infected patients with AIDS, has pathogenic properties associated with in-vivo virulence. Enzyme-linked immunosorbent assay and western blotting detected a more than 100 times higher frequency of antibodies to the mycoplasma in serum from HIV-1-infected patients with AIDS (40%) than from HIV-negative controls (0.3%). Serum from 20% of HIV-1-infected, symptom-free individuals also had M penetrans specific antibodies. The antibodies' major immunoreactivity was directed against P35 and P38, the two main lipid-associated membrane protein antigens of the organism. Patients attending sexually transmitted disease clinics had a low frequency of antibody (0.9%). None of 178 HIV-negative patients with different non-AIDS diseases, many associated with immune dysfunction and/or low white cell counts, tested positive for the antibodies. M penetrans, apparently not a commensal and not a simple opportunist, is uniquely associated with HIV-1 infection and AIDS.

AIDS-Related Opportunistic Infections↗

A comparison of case retrieval times: film versus picture archiving and communications systems.

One of the advantages that a picture archiving and communications system (PACS) is supposed to provide over a film-based operation is improved performance in retrieving images. Although it seems self-evident that this should be so, this experiment was intended to verify this and to provide some time comparisons for the two methods. The experiment consisted of randomly selecting ultrasound and computed tomography cases and determining how long it took to retrieve files at a PACS workstation or in person from the file room. To simulate actual retrieval volumes, a total of 40 cases from current to 6 months old, 20 cases from the past year, and 10 cases more than 1 year old was selected. Results indicate that PACS retrieval can indeed be faster than file room retrieval. However, the difference is less for recent cases than for older cases. For cases 6 or fewer months old, the workstation retrieval was approximately 2.5 minutes faster per case than the film file room. This time difference increased markedly when extended to the 1-year and older-than-1-year groups. This report details the results of this study and provides information about the reliability of the two archives.

Filing↗