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Biomedical subjects

D Li

Publications and source records attributed to D Li.

At least 919 records · Page 51Linked to original sources
PubMed1988

DNA probe technology for rapid detection of Haemophilus influenzae in clinical specimens.

In a previous study, we reported that a 5-kilobase Haemophilus influenzae DNA fragment involved in penicillin-binding protein expression could be used as a probe for specific detection of H. influenzae strains (F. Malouin and L. E. Bryan, Mol. Cell. Probes 1:221-232, 1987). Here, we report the ability of this probe to detect H. influenzae in clinical specimens. In a bacterial dot experiment, there was strong hybridization of the 32P-labeled probe to nonencapsulated and serotype a through f H. influenzae strains. The detection of H. influenzae in body fluids was then evaluated by using pooled human serum, urine, cerebrospinal fluid, and sputum as dilution media for H. influenzae, Haemophilus aegyptius, Haemophilus parainfluenzae, and Escherichia coli cells. At 65 degrees C, the probe hybridized to H. influenzae and H. aegyptius (greater than or equal to 10(5) cells) in all fluids. There was no hybridization with the E. coli negative control, and H. parainfluenzae hybridized when greater than or equal to 10(7) cells were used. Experiments performed at 73 and 80 degrees C permitted elimination of H. parainfluenzae hybridization. The detection of H. influenzae in 232 sputa from patients with respiratory tract infections was very specific (96 to 97%) and sensitive (74 to 100%) when the total time of the procedure was sufficient (6 to 24 h) and when the experiments were performed at 80 degrees C. In addition, the probe detected three of three and four of four H. influenzae-infected cerebrospinal fluids and blood cultures, respectively, and did not react with pneumococcus- or streptococcus-infected cerebrospinal fluids. Finally, by using a small-scale procedure, the probe rapidly detected H. influenzae in cerebrospinal fluid and sputum specimens (4 and 8 h, respectively). These results imply prompt diagnosis of H. influenzae infections caused by nonencapsulated and serotype a through f strains.

Body Fluids↗
PubMed1988

Multiple insertions and tandem repeats of origin-minus simian virus 40 DNA in transformed rat and mouse cells.

Stable simian virus 40 (SV40) transformation requires integration and expression of the early region of the SV40 genome. We have examined the amount and state of integrated viral DNA of SV40-transformed NIH 3T3 mouse and F2408 rat fibroblast lines generated by transfection with either wild-type or origin-defective SV40 DNA. A functional SV40 replication origin was not required for multiple inserts and partial-repeat structures to form in NIH 3T3 mouse transformants. In contrast, partial repeats in F2408 rat transformants were rare when the SV40 replication origin was intact and not detected at all when it was defective.

Animals↗
PubMed1988

Results of anterior corpus callosum section in 24 patients with medically intractable seizures.

Twenty-four medically refractory seizure patients, who did not qualify for excisional surgery, had anterior two-thirds corpus callosum section. Three to 11 years' postoperative follow-up suggests that this procedure can (1) lateralize a frontal lobe focus, which may lead to subsequent localized excision and (2) significantly reduce seizure frequency and severity in 75% of the patients without giving any permanent neurologic deficits. Patients with an ictal focus confined to one frontal lobe did best (8/8 improved), followed by patients with secondarily generalized seizures and multifocal bilateral foci (5/6 improved). Patients with mental retardation benefited less frequently (5/10 improved), but 4/4 from this group with ictal falls associated with Lennox-Gastaut syndrome did benefit. In this series, the improvements following the anterior partial section were lasting if present at 1 year of follow-up. Anterior corpus callosum section should be considered as a diagnostic (lateralizing) and therapeutic option in appropriately defined medically refractory patients who do not qualify for excisional surgery.

Adolescent↗
PubMed1987

Expression of five forms of microsomal cytochrome P-450 in primary cultures of rabbit hepatocytes treated with various classes of inducers.

In order to investigate the expression of five different forms of microsomal cytochrome P-450 including P-450 2 (P450IIBI), 3b (P450IIC3), 3c (P450IIIA4), 4 and 6 (P450IA2 and A1), hepatocytes were isolated from untreated rabbit and maintained in primary monolayer cultures in serum free modified Waymouth medium in the absence and in the presence of various classes of inducers including phenobarbital (PB), rifampicin (RIF), dexamethasone (DEX) and B-naphthoflavone (BNF). In untreated cultures the level of the various forms of P-450, determined by immunoblot with the use of specific antibodies, generally declined with time but at markedly different rates. In cultures treated with the inducers decline of the various forms was either unaffected, reduced, or even reversed, so that 96 hr after plating some of these forms appeared to be induced several-fold with respect to the untreated cultures. The forms 2 and 3c were co-induced by PB, RIF or DEX; as in vivo, BNF induced forms 4 and 6. Induction of forms 2, 3c, 4 and 6 was accompanied by stimulation of related monooxygenase activities, benzphetamine demethylase, progesterone 6B hydroxylase and benzpyrene hydroxylase and ethoxyresorufin deethylase, respectively. In all cases, induction was accompanied by an increased rate of de novo synthesis of the protein, determined by radio-immunoprecipitation assay with the use of specific antibodies on [3H]-Leu labeled cell lysate. Both induction and increased de novo synthesis were time- and inducer concentration-dependent. In cultures treated with RIF or BNF de novo synthesis of P-450 3c or of P-450 4 and 6 was correlated with the level of their specific mRNA quantitated from northern blots probed with either pLM3c-4.1 or pLM6.1, two plasmids containing inserted cDNA coding for P-450 3c or P-450 6, respectively. We conclude from these experiments that rabbit hepatocytes in primary monolayer cultures represent suitable models for studying regulation induction and pharmacological implications of the microsomal cytochromes P-450.

Animals↗
PubMed1987

Comparison of magnetic resonance imaging with somatosensory testing in MS suspects.

Two hundred patients suspected of having multiple sclerosis (MS), including 42 with progressive myelopathy and 11 with optic neuritis, were investigated with somatosensory evoked potentials (SEPs) and magnetic resonance imaging (MRI). Most had minimal neurological deficit, the mean Kurtzke scale being 2.65. There were 117 patients had two or more MRI lesions suggestive of MS, with a total of 527 lesions identified; 290 (55%) involved the somatosensory pathways, most commonly lying in the mid-periventricular region (thalamo-cortical radiations). There was good correlation between positive and negative MRIs and SEPs. The MRI was abnormal more frequently than the SEP, except in progressive myelopathy when both were abnormal with equal frequency. It is proposed that some cases of myelopathy in MS may be due to periventricular rather than spinal pathology. The morphology of the MRI lesion would favor conduction block not slowing of the SEP as being the prime abnormality. This appeared to be true of leg SEPs.

Adolescent↗
PubMed1987

Controlling for cerebral atrophy in positron emission tomography data.

This article outlines a simple method for estimating the degree of cerebral atrophy and subsequent experimental control for the effects of cerebral atrophy on regional metabolic values measured with positron emission tomography. It is suggested that rather than correcting derived values for atrophy, less potential error is introduced by experimentally controlling for regional atrophy when possible. By treating the derived metabolic rates and the measure of atrophy as two separate variables, it is also possible to establish whether there actually is a demonstrable relationship between the two measures.

Adolescent↗
PubMed1987

Lesions of the putamen: their relevance to dystonia.

Two patients with acquired dystonia were studied by computed imaging techniques and found to have lesions predominantly involving the putamen. The implications of these findings are discussed, and it is concluded that, for the genesis of dystonia, a relative increase of other inputs to the pallidum may be important, such as those from the caudate and subthalamic nuclei.

Adult↗
PubMed1986

Purification and partial characterization of prostate-derived growth factor.

A potent growth-promoting polypeptide, the prostate-derived growth factor (PrDGF), has been purified to apparent homogeneity from acid extracts of rat prostatic tissue using ion-exchange, reverse-phase, and gel-permeation chromatography. PrDGF migrates as a single protein-staining band in NaDodSO4/PAGE in precise correspondence to extractable PrDGF activity in nonstained NaDodSO4 gels. PrDGF is acid- and heat-stable but is sensitive to reduction or protease treatment. PrDGF is an acidic (pI 5.0) protein of approximately equal to 25 kDa in NaDodSO4/polyacrylamide gels and of approximately equal to 6-8 kDa in reduced NaDodSO4/polyacrylamide gels. PrDGF stimulates the linear incorporation of [methyl-3H]thymidine into normal rat kidney cells between 0 and 16 ng/ml. PrDGF appears to differ from other known growth factors in chemical composition and biological properties, suggesting that PrDGF is a previously undescribed growth factor.

Animals↗
PubMed1986

In vivo human tracheal pressure-area curves using computerized tomographic scans. Correlation with maximal expiratory flow rates.

In order to develop a simple technique to measure in vivo pressure-area (P-A) curves of the extrathoracic trachea in humans, we studied 14 normal male subjects. Valsalva and Mueller maneuvers were performed at FRC, and tracheal cross-sectional area (TXSA) was measured using computed tomography. Extrathoracic tracheal transmural pressure (TMP) was obtained as airway opening minus atmospheric pressure (Pat). Tracheal "compliance" (TC) was measured on the "inflation" limb of the P-A curve. Tracheal compliance was not a significant predictor of maximal expiratory flow rates, and TXSA at zero TMP was a significant predictor of peak expiratory flow rate but not of FEV1 or Vmax50. P-A curves showed an unexpected configuration characterized by a plateau or an increase in TXSA with TMP lower than -15 cm H2O. P-A curves obtained in 5 subjects using extrathoracic esophageal pressure as tracheal external pressure instead of atmospheric pressure did not show a plateau or an increase in TXSA with Mueller maneuvers. In these 5 subjects, TC using esophageal pressure rather than Pat did not aid in the prediction of flow. We conclude that extrathoracic tracheal external pressure is not Pat because this pressure is probably affected by transmission of pleural pressure to the cervical interstitial tissue as well as by the contraction of cervical accessory inspiratory muscles. Therefore, true tracheal compliance cannot be simply measured since it requires placement of an esophageal balloon.

Adult↗
PubMed1986

[Intraoperative cytodiagnosis of lung masses by fine needle aspiration].

The results of intraoperative cytodiagnosis by fine needle aspiration of 51 cases with lung masses are presented. The diagnosis of benign lesion in 17/51 cases and malignancy in 34/51 cases was established by cytology. The accuracy of this cytopathologic diagnosis was 100% in our series. In 34 cases with malignant lung tumors, the accuracy, as compared with histological diagnosis in cell type, was 84.2%. At thoracotomy, this approach is easy, safe and rapid and without any complication for undetermined pulmonary lesions. In some cases with a deep lesion, this approach has proved to be most useful to avoid a blind major pulmonary resection. We believe that intraoperative cytodiagnosis of lung masses by fine needle aspiration is a valuable supplementary diagnostic measure for pulmonary neoplasms.

Adenocarcinoma↗
PubMed1985

Synthesis and degradation of 3-methylcholanthrene-inducible cytochromes P-450 and their mRNAs in primary monolayer cultures of adult rat hepatocytes.

We used primary nonproliferating cultures of adult rat hepatocytes to investigate the regulation of P-450c and P-450d, immunochemically related protein products of separate cytochromes P-450 genes that are coinduced by 3-methylcholanthrene and related compounds. In cultures of hepatocytes prepared from untreated rats and incubated in media containing 3-methylcholanthrene, beta-naphthoflavone, 3,4,3',4'-tetrachlorobiphenyl, and Aroclor 1254 (a mixture of chlorinated biphenyls) there was a 5- to 15-fold accumulation of P-450c protein (quantitated by immunoblotting), accompanied by an increased rate of P-450c synthesis (measured as incorporation of [3H]leucine into immunoprecipitable protein) and an increased amount of P-450c mRNA hybridizable to a specific cloned cDNA (p210). In contrast, there were no increases in the concentration of P-450d protein, its rate of synthesis, or the amount of P-450d mRNA hybridizable to its specific cDNA (p72). Similarly, when "preinduced" hepatocytes (isolated from rats treated with Aroclor 1254) were incubated for 4 days in culture medium, the amount of P-450c, its rate of synthesis, and the amount of P-450c mRNA remained elevated, whereas synthesis of P-450d and the amount of P-450d mRNA fell precipitously to less than 10% of the initial values despite the presence or absence of Aroclor 1254 or of isosafrole in the medium. However, the loss of P-450d protein in these cultures was almost completely prevented when isosafrole was added to the culture medium and was partially prevented when safrole, Aroclor 1254, and 3,4,5,2',4',5'-hexachlorobiphenyl, but not 3-methylcholanthrene, beta-naphthoflavone, or 3,4,3'4'-tetrachlorobiphenyl, were in the culture medium. Moreover, in similar cultures of "preinduced" hepatocytes that were pulse-labeled with [3H]leucine, the presence of isosafrole in the culture medium extended the apparent half-life for loss of radioactivity in immunoprecipitable P-450d to a value of 72 h (3-fold longer than in standard medium) but was without effect on the rate of disappearance of radiolabeled P-450c. We conclude that control of P-450d degradation is an important factor in the regulation of this hemoprotein and that induction of P-450c and P-450d proceed by separate pathways that are spontaneously divorced under standard conditions for primary culture of adult rat hepatocytes.

Animals↗
PubMed1985

Nonprotein sulfhydryl compounds in canine gastric mucosa: effects of PGE2 and ethanol.

By use of an in vivo canine chambered stomach preparation in which the gastric mucosa was partitioned into two equal halves, the effect of topical 16,16-dimethyl PGE2 (DMPGE2) (1 microgram/ml of perfusate) and 8% and 40% ethanol on tissue levels of nonprotein sulfhydryl compounds was assessed. Both DMPGE2 and 8% ethanol significantly increased (P less than 0.005) mucosal levels of nonprotein sulfhydryls when compared with corresponding mucosa bathed with saline alone. In contrast, mucosa bathed with 40% ethanol showed significantly decreased levels. If mucosa was bathed with DMPGE2 or 8% ethanol prior to exposing the stomach to 40% ethanol, this depletion in sulfhydryl compounds was not observed. Since other experimental observations have shown that exogenously administered prostaglandins and mild irritants (such as low-dose alcohol) can prevent gastric mucosal damage by necrotizing agents (such as high-dose alcohol), our findings are consistent with the hypothesis that nonprotein sulfhydryls may play a role in mediating gastric mucosal protection.

16,16-Dimethylprostaglandin E2↗
PubMed1985

Mechanism of action of a suppressor-activating factor (SAF) produced by a human T cell line.

We previously described a potent suppressor-activating factor (SAF) produced constitutively by a 6-thioguanine-resistant mutant of the human T cell line CEM. In the present study, we investigated the mechanism of action of SAF. After a brief (4- to 18-hr) exposure to SAF at 37 degrees C, T lymphocytes (either unseparated, or purified OKT4+ and OKT8+ subpopulations), but not B lymphocytes, suppressed allogeneic and syngeneic T cells in co-culture experiments, apparently via the release of a suppressor activity. The total T cell-released suppressor activity (TRSA) accumulated after 3 days culture post-treatment was about 100- to 500-fold higher than the original suppressor activity (SAF) added to trigger the release. Arresting protein or DNA synthesis, or even killing the cells did not affect the release of TRSA by T lymphocytes, but lowering the incubation temperature to 4 degrees C reduced it drastically. Pre-treatment of T lymphocytes with the metabolic inhibitor, sodium azide, or the adenylate cyclase stimulator, prostaglandin E2, or the addition of exogenous dibutyryl cAMP, all suppressed the release of TRSA. The presence of monoclonal antibody OKT3, but not OKT4 or OKT8, enhanced the release of TRSA. The presence of OKT11 blocked the release of SAF. The functional characteristics of TRSA appeared to be identical to those of SAF. However, unlike SAF, interaction of T lymphocytes with TRSA triggered only marginal enhancement of suppressor activity. In addition, the kinetics of the suppression mediated by SAF showed a much larger increment as a function of time than that mediated by TRSA. Taken together, the data suggest that SAF might represent an activated form of SAF, and that the continuous activation of SAF by lymphocytes in culture may account for its high potency in suppressing T cell proliferation in vitro.

Binding Sites↗