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L Chen

Publications and source records attributed to L Chen.

At least 1,495 records · Page 83Linked to original sources
PubMed1985

ATP is essential for protein translocation into Escherichia coli membrane vesicles.

The energy requirement for translocation of alkaline phosphatase and the outer membrane protein OmpA into Escherichia coli membrane vesicles was studied under conditions that permit posttranslational translocation and, hence, prior removal of various components necessary for protein synthesis. Translocation could be supported by an ATP-generating system or, less well, by the protonmotive force generated by D-lactate oxidation; the latter might act by generating ATP from residual bound nucleotides. However, when protonmotive force inhibitors were used or when ATP was further depleted by E. coli glycerol kinase, D-lactate no longer supported the translocation. Furthermore, ATP could still support protein translocation in the presence of proton uncouplers or with membranes defective in the F1 fraction of the H+-ATPase. We conclude that ATP is required for protein translocation in this posttranslational system (and probably also in cotranslational translocation); the protonmotive force may contribute but does not appear to be essential.

Adenosine Diphosphate↗
PubMed1985

Cell to cell transmission of the "priming" effect on the induction of human fibroblast interferon.

Human fibroblast cells remain primed for IFN-beta 1 synthesis for at least 18 days after the removal of IFN-beta 1, to the same extent as cells primed shortly before induction. The maximal effect of priming is observed in cells that are treated with IFN at low cell density and are subsequently allowed to undergo several divisions. This observation suggests that the information for priming is propagated from cell to cell upon cell division. A diminished priming effect is observed after cells undergo more than three divisions in culture, suggesting a dilution of the mediator responsible for the transmission of the effect. It was not possible to demonstrate intercellular communication of the priming effect by a mediator found in the medium or by cell to cell contact. Observation that priming persists in cells for 18 days in the absence of detectable levels of IFN in the medium, but virus-resistance declines after three days supports the previous suggestion for different mechanisms involved in these two IFN-induced phenomena.

Cell Communication↗
PubMed1985

The influence of halothane and isoflurane on pulmonary collateral ventilation.

The effects of halothane and isoflurane on hypocapnic increases in pulmonary collateral resistance were studied in dogs. A bronchoscope with a double lumen catheter in the suction port obstructed a peripheral airway and allowed gas to flow out of the isolated segment of lung only via collateral channels. The collateral gas flow (Vcoll) was measured with a flowmeter and delivered through one lumen of the catheter, while the other lumen measured distal pressure (Pb). At FRC, the resistance to collateral ventilation (Rcoll) was calculated as Rcoll = Pb/Vcoll. The rest of the lung was ventilated with air, while air (hypocapnia), 10% CO2 in air, or air and halothane or isoflurane were delivered to the isolated segment. A measurement of resistance was made after 4 min of test gas flow. For each segment, when air replaced 10% CO2, the average increase in Rcoll was calculated and called Rmax. When 10% CO2 in air was infused into segments the mean Rcoll (n = 50) was 0.0196 +/- 0.0022 cmH2O X ml-1 X min. This increased to 0.0285 +/- 0.0031 cmH2O X ml-1 X min (mean +/- E) when air was infused, a mean increase in resistance of 52 +/- 3%. When halothane or isoflurane was added to air the hypocapnic increase in Rcoll was attenuated with a 50% decrease at 1.3% (1.4 MAC and 0.8 MAC, respectively). These two inhalational anesthetics reduce active changes in the flow resistance to collateral ventilation. When collateral resistance acts to adjust ventilation perfusion deviations, this action of halothane and isoflurane may make this regulation less effective.

Airway Resistance↗
PubMed1985

Hypoxic pulmonary vasoconstriction is not potentiated by repeated intermittent hypoxia in closed chest dogs.

Hypoxic pulmonary vasoconstrictor (HPV) responses were measured with repeated intermittent hypoxic challenges in eight non-traumatized closed chest dogs anesthetized with pentobarbital. The right lung was ventilated continuously with 100% O2 while the left lung was either ventilated with 100% O2 (control) or ventilated with a gas mixture containing 3-4% O2 (hypoxia). Mean per cent left lung blood flow for all four normoxic periods was 43.1 +/- 1.5% (mean +/- SE) of the total blood flow by the SF6 excretion method and 40.8 +/- 1.1% by the differential CO2 excretion method, corrected for the Haldane effect. With hypoxic ventilation, flow diversion from the hypoxic lung was maximal with the first exposure and did not change subsequently with a total of four alternating exposures to normoxia and hypoxia. Flow diversion during hypoxia was approximately 50.5 +/- 2.4% by the SF6 method and 50.3 +/- 3.5% by the VCO2 method. This result contrasts with the increasing flow diversion response with intermittant hypoxic exposure that has been reported in animals exposed first to thoracotomy and surgical dissection. It is concluded that in the absence of surgical trauma the initial response to hypoxia is maximal and is not potentiated by repeated hypoxic stimulation.

Anesthesia, Intravenous↗
PubMed1985

Alkaline phosphatase and OmpA protein can be translocated posttranslationally into membrane vesicles of Escherichia coli.

We previously described a system for translocating the periplasmic enzyme alkaline phosphatase and the outer membrane protein OmpA into inverted membrane vesicles of Escherichia coli. We have now optimized and substantially improved the translocation system by including polyamines and by reducing the amount of membrane used. Under these conditions, efficient translocation was seen even posttranslationally, i.e., when vesicles were not added until after protein synthesis was stopped. This was the case not only with the OmpA protein, which is synthesized by free polysomes and hence is presumably exported posttranslationally in the cell, but also with alkaline phosphatase, which is synthesized only by membrane-bound polysomes and has been shown to be secreted cotranslationally in the cells. Prolonged incubation rendered the precursors inactive for subsequent translocation. Posttranslational translocation was impaired, like cotranslational translocation, by inhibitors of the proton motive force and by treatment of the vesicles with protease. Since it appears that E. coli can translocate the same proteins either cotranslationally or posttranslationally, the cotranslational mode may perhaps be more efficient, but not obligatory, for the secretion of bacterial proteins.

Alkaline Phosphatase↗
PubMed1985

[A simple method for performing yeast carbohydrate assimilation test].

A modified auxanographic method was employed to perform carbohydrates assimilation test for 25 strains of yeasts representing the following 8 species: Candida albicans, C. glabrata, C. krusei, C. parapsilosis, C. stellatoides, C. tropicalis, Cryptococcus neoformans, and Rhodotorula glutinis. The results obtained were analyzed by comparing with the conventional auxanographic method, and it was found that the modified method was more accurate, rapid and easy in reading. The modified method was obviously easier in manipulation and more economic than that of Wickerham's broth technique and yeast assimilation agar slant technique.

Candida albicans↗
PubMed1984

A human IFN-beta 1 gene deleted of promoter sequences upstream from the TATA box is controlled post-transcriptionally by dsRNA.

Induction of IFN-beta 1 RNA was studied in the mouse cell line SR117-21E transformed by a BPV episome containing the human IFN-beta 1 gene deleted of promoter sequences upstream from position -40. Nuclei isolated from these cells synthesize constitutively IFN-beta 1 RNA from the partially deleted promoter. The IFN-beta 1 RNA synthesized by nuclei of uninduced SR117-21E cells is similar to that made by nuclei of poly(rI):(rC)-induced cells, but does not accumulate and hence no IFN is produced unless the cells have been treated either by ds RNA or by cycloheximide. We conclude that the IFN-beta 1 gene has, in addition to the transcription control due to upstream promoter sequences, an additional post-transcriptional control acting on mRNA accumulation and linked to sequences close to the TATA box and RNA start site. Both controls are relieved by ds RNA.

Animals↗
PubMed1984

Nucleotide sequence of a multiple-copy gene for the B protein of photosystem II of a cyanobacterium.

Chloroplast photogene 32 codes for the 32-kilodalton triazine herbicide-binding protein at the B site of electron transport in photosystem II of the photosynthetic apparatus-its product is the B protein and the gene is accordingly designated ps2B here. The cyanobacteria Anacystis nidulans R2, Fremyella diplosiphon, and Nostoc sp. MAC each contain several copies of ps2B. The sequence of one copy of ps2B from Fremyella, ps2B-1, has been determined. The longest open reading frame would code for a protein of 360 amino acids. Although the deduced amino acid sequence of ps2B-1 is highly homologous overall to that of the corresponding spinach protein [Zurawski, G., Bohnert, H. J., Whitfeld, P. R. & Bottomley, W. (1982) Proc. Natl. Acad. Sci. USA 79, 7699-7703] and, excluding neutral substitutions, the homology is 95% for an internal segment of 309 amino acids, there are a number of nonneutral amino acid substitutions. Most of the differences in net charge and polarity occur in the first 20 amino acids at the amino terminus and in the amino acid composition at the carboxyl terminus. The nucleotide sequences are 76% homologous overall. Conserved sequences resembling prokaryotic "-10" and "-35" regions are found at remarkably similar positions in the spinach and F. diplosiphon sequences although the surrounding sequences show only occasional homologies.

Journal Article↗
PubMed1984

Efficient constitutive production of human fibroblast interferon by hamster cells transformed with the IFN-beta 1 gene fused to an SV40 early promoter.

The coding sequence of the human interferon (IFN)-beta 1 gene, fused 60 bp downstream from the RNA start site of the SV40 early gene, was transfected into dihydrofolate reductase (DHFR)-deficient Chinese hamster ovary (CHO) cells together with a selectable DHFR gene. Most transformants continuously secreted IFN-beta 1 into the medium. Induction did not stimulate expression of the fused SV40-IFN-beta 1 gene. The role of the SV40 promoter was verified by transforming cells with the unmodified human IFN-beta 1 gene, or by the IFN-beta 1 coding region fused to another poly(rI):(rC)-inducible gene. In these cases, the transformants showed strictly inducible (not constitutive) IFN secretion. By selection for methotrexate resistance, CHO clones with a 10-20-fold amplification of the SV40-IFN-beta 1 DNA were obtained. Such clones constitutively produce up to 350,000 units IFN/ml per 10(6) cells/24 hr, i.e., over 10 times more than fully induced human fibroblasts. In continuous culture with daily changes of medium, accumulation of IFN-beta 1 is constant at a rate of 300,000 molecules per cell/hr. Batches of up to 16 mg of IFN-beta 1 produced by the transformed CHO cells were purified to homogeneity by affinity chromatography on monoclonal antibodies. This IFN appears identical in size, activity, and immunospecificity to the native human IFN-beta 1 glycoprotein.

Animals↗
PubMed1984

Time course and responses of sustained hypoxic pulmonary vasoconstriction in the dog.

The stability of the pulmonary blood pressure and flow response to alveolar hypoxia (hypoxic pulmonary vasoconstriction or HPV) was studied in six pentobarbital anesthetized, mechanically ventilated open-chested dogs. Aortic and left pulmonary artery blood flows; systemic and pulmonary arterial, central venous, left atrial, and airway pressures; hemoglobin; arterial and mixed venous blood gases were measured. The right lung was ventilated continuously with 100% oxygen, while the left lung was ventilated alternately with 100% O2 ( prehypoxia control phase), an hypoxic gas mixture containing 4% O2, 3% CO2, balance N2 for 4 h, or 100% O2 (post-hypoxia control phase). Hypoxic ventilation of the left lung resulted in an immediate and sustained decrease in left lung blood flow (QL%) from 39.0 +/- 1.8% (mean +/- SE) to 9.9 +/- 3.6% at 15 min of hypoxic ventilation. QL% remained decreased and did not vary significantly during the 4 h of hypoxia. Venous admixture correspondingly was increased and PaO2 decreased by hypoxic ventilation and did not vary significantly during the 4 h of hypoxia. All variables returned to control levels upon reestablishing ventilation with 100% O2. While the maximal reduction in QL% with left lung hypoxic ventilation was identical to that observed during atelectasis previously in our laboratory, the time course of the response was different. The response to hypoxia was maximal by 15 min, however, QL% decreased more slowly during atelectasis, where the maximal reduction was observed by 60 min. The present study therefore demonstrated that hypoxic ventilation of the left lung yielded an immediate and sustained decrease in left lung blood flow for 4 h. The stability of the HPV response probably was accounted for by the lack of such confounding factors as respiratory alkalosis, severe systemic hypoxemia, and increased cardiac output.

Analysis of Variance↗
PubMed1984

Influence of the method of re-expansion of atelectatic lung upon the development of pulmonary edema in dogs.

An endobronchial tube was used to separately ventilate each lung in anesthetized dogs, and absorption atelectasis was then induced in the left lung and maintained for 3 1/2 h. The left lung was partially re-expanded by ventilation at normal tidal volumes for 10 min, and then completely re-expanded by manual hyperinflation. This delayed re-expansion was associated with an extravascular water content significantly greater than that of the right lung. A previous study demonstrated no increase in the ratio of left to right lung water content after 3 h of atelectasis alone and a significant increase after immediate hyperinflation. Delayed re-expansion produces significantly different results, and it is therefore concluded that the method of lung re-expansion influences the development of pulmonary edema after atelectasis.

Animals↗
PubMed1984

Clinical and immunopathologic study of mesangial IgM nephropathy: report of 41 cases.

Forty-one out of 408 cases (or 10%) of primary glomerular disease had diffuse fine granular to arc-like short linear mesangial deposits of IgM by direct immunofluorescence. The IgM deposition was accompanied by C1q and/or C4 in the same locality in 29 cases, by C3 in 10, and by trace amounts of IgA in 6. Properdin-factor B was not detected. Fine granular electron dense deposits of low density were detected in the mesangium in all 41 cases by electron microscopy, usually as a discrete granular or arc-like pattern beneath the mesangial glomerular basement membrane and correlated well with the immunofluorescence findings. An immune complex disease with complement activation via the classical pathway is suggested. The ages of the patients varied from 2 to 58 years (average 23.8 years). A male predominance of 2.2:1 was identified. Serum IgM level was elevated in 46.7% of the cases. The majority (87.8%) of the cases manifested a nephrotic syndrome or relapse at time of biopsy, and the remaining cases experienced persistent or intermittent proteinuria. Among the 36 nephrotic patients, 22 cases (61.1%) demonstrated complete remission with steroid therapy, 9 cases (25%) were resistant, and 5 cases (13.9%) had partial remission. Complete and partial remissions were later achieved with cytotoxic drugs or methylprednisolone pulse therapy in 3 and 4 cases respectively in the steroid resistant patients. Frequent relapses occurred during the course in 22 out of 32 cases (68.8%) who had experienced complete or partial remission. Follow-up study after biopsy demonstrated that sustained complete remission was achieved with prednisolone with or without cytotoxic drugs and pulse therapy in only 14 (42.4%) of the 33 nephrotic cases who had been followed up for longer than 6 months, and six of them had had previous relapses. Pathologically, 56.1% of the patients showed mild to moderate increase in mesangial matrix and cellularity. Focal and segmental sclerosis was demonstrated in four cases (9.8%). However, minimal glomerular change was also common (34.1%). The patients with minimal change seemed to have a higher complete remission rate than patients with more evident glomerular alterations, although the difference was not statistically significant. This clinical and immunopathological study suggests that mesangial IgM nephropathy is an important disease in Taiwan, with a variable response to treatment and frequent relapses.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗