Biomedical subjects

H C Wu

Publications and source records attributed to H C Wu.

At least 91 records · Page 5Linked to original sources

PubMed1993

Characterization of seven newly established nasopharyngeal carcinoma cell lines.

BACKGROUND: Nasopharyngeal carcinoma (NPC) has a relatively high incidence in Chinese living in Taiwan, Hong Kong, Singapore, and South China. To better understand this cancer, we have established several new NPC cell lines. EXPERIMENTAL DESIGN: We collected biopsy specimens from suspected NPC patients and divided each specimen into two parts: the first part was fixed for routine histopathologic examination, and the other part was put into culture medium for primary culture. Once the cell lines were established, they were extensively characterized. RESULTS: Seven NPC cell lines were established, and all have been passaged more than 100 times. Two lines were derived from keratinizing carcinomas and five from undifferentiated carcinomas. Electron microscopic examination revealed that both dark and light tumor cells contained intermediate filaments with clear desmosome formation. The average doubling time ranged from 10.7 to 16.3 hours. Karyotypic analysis showed multiple chromosome abnormalities with the average chromosome number between 84 and 95. Colony forming efficiency in soft agar was 18-42%. All cell lines could induce solid tumor mass formation when transplanted into nude mice, and the histopathological findings showed two keratinizing and five nonkeratinizing carcinomas. All cell lines contained less acidic keratin polypeptides than basic keratin polypeptides. Strong expression of vimentin in each single cell of all cell lines was also observed. The oncosuppressor retinoblastoma gene in each cell line showed no remarkable abnormality, but retinoblastoma protein was abnormally expressed in some interphase cells. The oncogenes, erbB and c-fgr, were both normally expressed. While the c-myc oncogene in all cell lines was overexpressed when compared with the Burkitt's lymphoma Raji Cell line, the c-myc DNA sequence in each cell line showed neither amplification nor rearrangement. CONCLUSIONS: The newly established seven NPC cell lines have been well characterized, and all showed overexpression of c-myc oncogene. The oncosuppressor retinoblastoma gene revealed no remarkable rearrangement, but its protein product was abnormal in certain interphase cells of each cell line.

Adult↗

PubMed1993

Removal of renal cell carcinoma extending into the right atrium using cardiopulmonary bypass, profound hypothermia and circulatory arrest.

A 65-year-old man presented with renal cell carcinoma of the right kidney with a tumor thrombus extending up the vena cava to the right atrium. Cardiopulmonary bypass, profound hypothermia and total circulatory arrest were used to create a bloodless field for excision of the renal cell carcinoma and its tumor thrombus. Acute respiratory failure and deep jaundice developed after the operation and the patient was transferred to the intensive care unit for critical care. After respiratory therapy and nutritional support, the liver function was restored. The endotracheal tube was weaned one month later. The patient has had total resolution of all symptoms and there is no evidence of tumor recurrence of distant metastases after 6 months follow-up.

Aged↗

PubMed1993

[Analgesic effect of tramadol HCL in ESWL].

Since its introduction, extracorporeal shock-wave lithotripsy (ESWL) has become the treatment of choice in patients with urinary calculi. But the pain during ESWL is intolerable for many patients. Tramadol HCL resembles morphine in that it depresses motor and sensory responses of the spinal nociceptive system by a spinal and a supraspinal action. The side effects of tramadol are less than morphine. A prospective study was performed to determine the effect of tramadol in ESWL for the patients with urinary tract calculi. Ninety patients were randomized divided into three groups. Group A 40 patients (male:female = 31:9) received oral tramadol HC1 100 mg; group B 17 patients (male:female = 12.5) received multi-vita; group C 33 patients (male:female = 26:7) received analgesic (contained aspirin 399 mg and codeine phosphate 15 mg). The patients took the drugs 30 minutes before ESWL. The patients with renal calculi > 2 cm in diameter or ureteral calculi > 1 cm in diameter were excluded. The lithotriper used in our hospital is Siemens Lithostar. According to the description of the patients postoperatively, the pain intensity during ESWL was identified with Verbal Scale and Visual Analogue Scale (VAS). The statistical method is one-way ANOVA. Statistical difference is significant when p < 0.05. The mean age of patients group A is 46.5 +/- 17.1, group B 45.5 +/- 14.4, and group C 50.5 +/- 14.6. The mean diameter of urinary calculi of group A is 0.98 +/- 0.41 cm, group B 1.09 +/- 0.33, and group C 1.06 +/- 0.43.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1992

Deletion of internal twenty-one amino acid residues of Escherichia coli prolipoprotein does not affect the formation of the murein-bound lipoprotein.

Mutation pgsA affecting the phosphatidylglycerol phosphate synthesis is lethal for all but certain E. coli strains such as strains deleted for the lpp gene or strains containing unmodifiable prolipoprotein like lppD14. Strain SD312 pgsA3 is tolerant to pgsA mutation, which suggests the lpp alleles in strain SD312 pgsA3 and its parental strain SD12 may be defective. DNA sequence analysis of the lpp genes in Escherichia coli strains SD12 and SD312 pgsA using asymmetric polymerase chain reaction showed that the lpp alleles in these two strains contained a 63 base pair deletion corresponding to the 37th to 57th codons of the wild-type lpp gene. [3H]Palmitate labeling of strains SD12 and SDS312 showed that the mutant lipoprotein in SD12 strain was modified with lipid, while the prolipoprotein in SD312 was not modified. The shortened mature lipoprotein in SD12 and the lipid-modified prolipoprotein in globomycin-treated SD12 were found to be covalently attached to the peptidoglycan, while the unmodified prolipoprotein in SD312 did not form significant amounts of murein-bound lipoprotein.

Alleles↗

PubMed1992

Staphylococcal ADP-ribosyltransferase-sensitive small G protein is involved in brefeldin A action.

An early event in the action of brefeldin A (BFA) is the dissociation of beta-coat protein (beta-COP) from the Golgi membrane. We have recently shown that staphylococcal ADP-ribosyltransferase (epidermal cell differentiation inhibitor (EDIN)), which specifically modifies a small G protein, rho, mimics the action of BFA and disassembles the Golgi apparatus in Vero cells (Sugai, M., Chen, C-h., and Wu, H. C. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 8903-8907). Three independent BFA-resistant cell lines (BER-40 from Vero cells, PtK1, and MDCK) showed cross-resistance to EDIN regarding the release of the beta-COP from the Golgi membrane by EDIN or BFA. BFA as well as EDIN induced disassembly of the actin microfilaments in Vero cells, and they both failed to induce the disassembly of actin microfilaments in BER-40, PtK1, and MDCK cells. BFA inhibited protein secretion in Vero cells but not in BFA-resistant cell lines, whereas EDIN did not inhibit protein secretion in either Vero or other cell lines. AlF-4 inhibited the effect of EDIN as well as that of BFA on the distribution of the beta-COP. These results suggest that an EDIN-sensitive rho protein together with trimeric and other small G protein(s) is involved in the regulation of the assembly of coated vesicles and vesicular transport in the Golgi apparatus.

Animals↗

PubMed1992

Bacterial ADP-ribosyltransferase with a substrate specificity of the rho protein disassembles the Golgi apparatus in Vero cells and mimics the action of brefeldin A.

Epidermal-cell differentiation inhibitor (EDIN) is an exoenzyme produced by Staphylococcus aureus that catalyzes the ADP-ribosylation of rho proteins, members of the small GTP-binding protein family. In this study we demonstrate that EDIN induces a rapid morphological change in the Golgi structure of monkey kidney Vero cells that is similar to the changes elicited by brefeldin A (BFA). Treatment of Vero cells with EDIN resulted in a rapid disappearance of N-7-(4-nitrobenzo-2-oxa-1,3-diazole)-6-aminocaproylsphingosine, a 110-kDa protein (beta-COP, coat protein), and mannosidase II from the Golgi structure. Lower doses of EDIN and BFA had a synergistic effect on the redistribution of the Golgi markers. The similarities in the effects of EDIN and BFA in Vero cells also include the EDIN- or BFA-mediated protection of Vero cells from ricin cytotoxicity and prevention of the effects of EDIN or BFA on the distribution of Golgi markers by the pretreatment of Vero cells with guanosine 5'-[gamma-thio]triphosphate or forskolin. Incubation of a Vero-cell homogenate with [32P]NAD+ and EDIN in vitro resulted in the appearance of a labeled band with an apparent molecular mass of 22 kDa. The morphological change of the Golgi structure induced by EDIN was inhibited by nicotinamide, an inhibitor of EDIN-catalyzed ADP-ribosylation. Thus these data suggest that a rho protein is involved in the membrane trafficking between the Golgi and the endoplasmic reticulum of Vero cells and that this rho protein may be a target shared by EDIN and BFA.

4-Chloro-7-nitrobenzofurazan↗

PubMed1992

Alterations of the carboxyl-terminal amino acid residues of Escherichia coli lipoprotein affect the formation of murein-bound lipoprotein.

Mutations in the Escherichia coli lpp gene resulting in the alterations of the COOH-terminal region of the lipoprotein have been isolated by oligonucleotide-directed mutagenesis. As might be expected, substitution of Lys78 with Arg78 completely abolished the formation of murein-bound lipoprotein. Each of the following single amino acid substitutions did not significantly affect the formation of bound-form lipoprotein: Asp70 to Glu70 or Gly70; Lys75 to Thr75; and Tyr76 to His76, Ile76, or Leu76. In contrast, mutational alterations of Tyr76 to Cys76, Gly76, Asn76, Pro76, or Ser76 resulted in a reduction of the bound-form lipoprotein to levels of 14-32% of that in the wild-type strain. A common feature of these lpp COOH-terminal mutations affecting the formation of bound-form lipoprotein is the presence of a beta-turn secondary structure at the COOH-terminal region of all these mutant lipoproteins. In addition, substitution of Tyr76 to Asp76 or Glu76, and Arg77 to Asp77 or Leu77 also resulted in a reduced formation of the bound-form lipoprotein. These results suggest that the formation of murein-bound lipoprotein requires a COOH-terminal Lys residue and a positively charged COOH-terminal region. Furthermore, a beta-turn secondary structure in the COOH-terminal random coil region interferes with the attachment of the lipoprotein to the peptidoglycan.

Amino Acid Sequence↗

PubMed1992

Neither lipid modification nor processing of prolipoprotein is essential for the formation of murein-bound lipoprotein in Escherichia coli.

The relationship between the modification and processing of prolipoprotein and the formation of murein-bound lipoprotein has been investigated using Escherichia coli mutants altered in the signal sequence of prolipoprotein and an E. coli strain producing OmpF-Lpp hybrid protein. The glyceride-modified prolipoprotein in mutant lppT20 and in globomycin-treated wild-type strain were covalently attached to the peptidoglycan. Likewise, the unmodified prolipoproteins in mutants lppL20, lppV20, and lppG21 were attached to the peptidoglycan. The OmpF-Lpp hybrid protein that is processed but not modified with lipid due to the absence of the cysteine-containing modification site in the hybrid protein was also covalently linked to the peptidoglycan. These results indicate that neither lipid modification nor the processing of prolipoprotein is essential for the formation of murein-bound lipoprotein in E. coli. In contrast, introduction of a charged amino acid residue such as Asp or Arg at the 14th position of prolipoprotein affected not only the lipid modification and processing of the mutant prolipoprotein but also the formation of murein-bound lipoprotein. Replacement of the Gly14 with Glu or Lys partially affected the lipid modification and processing of prolipoprotein; the peptidoglycan of the lppE14 and lppK14 mutants contained a reduced amount of mature lipoprotein but no mutant prolipoprotein. In addition, lpp mutants A20I23I24 and A20I23K24 were found to be defective in both lipid modification/processing of prolipoprotein and the formation of murein-bound lipoprotein. The defective formation of murein-bound lipoprotein in the latter mutants may be related to an alteration in the secondary structure at the modification/processing site of the mutant prolipoproteins.

Amino Acid Sequence↗

PubMed1992

Nucleotide sequence of the Staphylococcus aureus signal peptidase II (lsp) gene.

The lsp gene encoding prolipoprotein signal peptidase (signal peptidase II) of Staphylococcus aureus was cloned by screening a genomic library for plasmid clones capable of complementing a conditionally lethal lsp allele of Escherichia coli. E. coli cells carrying one of five overlapping clones exhibited increased resistance to globomycin. The nucleotide sequence of the S. aureus lsp gene was determined. The deduced amino acid sequence of the signal peptidase II of S. aureus suggests that this enzyme has a hydropathy profile very similar to those of E. coli, Enterobacter aerogenes and Pseudomonas fluorescens. Comparison of the primary structures of this enzyme from these four distinct bacterial species reveals three highly conserved domains in proteins which have a low degree of overall sequence homology. Unlike the lsp genes from the Gram-negative bacteria, the lsp gene in S. aureus is not flanked by x-ileS and orf149-orf316 as found in E. coli, Ent. aerogenes, and P. fluorescens.

Amino Acid Sequence↗

PubMed1992

Serum prostate specific antigen levels in mice bearing human prostate LNCaP tumors are determined by tumor volume and endocrine and growth factors.

The ability of prostate-specific antigen (PSA) to predict tumor volume and stage in patients with prostate cancer would be improved if factors regulating its production and clearance were better defined. A thorough understanding of the pharmacokinetics (regulation of production, metabolism, and excretion) of PSA has been precluded, however, by the absence of an in vivo animal model. The purposes of this study are to develop a murine model for evaluating PSA pharmacokinetics in vivo and to assess factors that influence PSA production in vitro. The human prostate cancer cell line, LNCaP, was chosen because it is androgen sensitive and PSA positive. Although LNCaP cells are usually nontumorigenic when inoculated s.c. in athymic mice, coinoculation of 1 x 10(6) LNCaP cells with 1 x 10(6) human bone fibroblasts reliably produces PSA-secreting carcinomas. This LNCaP model provides accurate correlation between tumor volume and serum PSA levels (r = 0.94) and demonstrates that tumor volume and androgens are codeterminants of circulating PSA levels. Following castration, serum PSA levels decrease rapidly up to 8-fold and increase up to 20-fold following androgen supplementation, without detectable castration-induced tumor cell death or concomitant changes in tumor volume. Serum PSA levels increase 0.24 ng/ml/mm3 of tumor, which is approximately 5-fold less than that estimated for humans. Most likely this reduced PSA index (PSA:tumor volume ratio) results from a 7-fold faster clearance of PSA in athymic mice than in humans; other than this shorter half-life, PSA elimination in the murine model appears similar to that in humans, with both following first-order kinetics characteristic of a two-compartment model. Interestingly, following prolonged growth (greater than 21 days) in castrate hosts, LNCaP tumors are capable of adapting to an androgen-deprived environment whereby LNCaP tumors regain the ability to secrete PSA in amounts similar to the precastrate state. In LNCaP cells, androgens increase PSA mRNA levels 4-fold in vivo and in vitro. PSA mRNA expression is also altered by various growth factors. Changes in PSA production induced by androgens and growth factors do not always parallel changes in LNCaP cell growth rate induced by these factors, suggesting that PSA production occurs independently of cell growth rate and may be influenced by various interrelated factors, including hormonal and stromal milieu. Observations from this murine model suggest that androgens and tumor volume are independent determinants of serum PSA levels and imply that decreases in circulating PSA following antiandrogen therapy may not always reflect a corresponding reduction in tumor volume.

Animals↗

PubMed1992

Human prostate cancer model: roles of growth factors and extracellular matrices.

A human prostate cancer model was established by inoculating a prostate specific antigen (PSA)-producing LNCaP cell line with either prostate or bone fibroblasts. Alternatively, this human prostate cancer model can also be established by inoculating LNCaP cells with growth factor(s) (GFs) and extracellular matrix (ECM) immobilized on Gelfoam. The resulting LNCaP tumors were used to evaluate PSA production and excretion in athymic hosts. This model was also employed to examine the biochemical nature of mesenchymal cell-derived growth-promoting protein(s) and to assess the efficacy of potential chemotherapeutic agents. Because of the propensity of human prostate cancer to metastasize to the bone, this study defined a 1.0 M NaCl-eluted fraction, MS1, from the conditioned medium of a bone stromal cell line (MS) by heparin-affinity column chromatography. The growth-promoting activity was assayed both in vivo (e.g., tumor formation) and in vitro (e.g., soft agar colony formation). We found that the growth-promoting activity was trypsin- and heat-sensitive, and partially degraded by acid and dithiothreitol. Immunochemical studies indicated that the polyclonal antibody raised against MS1 blocked the growth-promoting effect elicited by the bone-conditioned media. This growth-promoting factor was found to be immunochemically dissimilar to KGF, HGF, and bFGF. However, addition of bFGF, HGF and NGF, but not aFGF, TGF beta, IGF1, IGF2, PDGF, EGF, TGF alpha and KGF, stimulated anchorage-independent growth of prostate cells, a condition closely parallel to tumor formation in vivo. We found that the MS1 fraction also contained fibronectin and tenascin but not laminin or collagen IV.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Isolation and characterization of a brefeldin A-resistant mutant of monkey kidney Vero cells.

Brefeldin A (BFA) is a fungal antibiotic which disrupts protein transport between the endoplasmic reticulum and the Golgi. A BFA-resistant mutant of monkey kidney Vero cells, BER-40, which exhibited about a 90-fold increase in the LD50 of BFA (5.2 ng/ml for Vero cells versus 460 ng/ml for BER-40 cells), has been isolated. The increased resistance of BER-40 cells toward BFA was also manifested in a greatly reduced inhibition of protein secretion by BFA in the mutant and a lack of protection by BFA of the mutant cells from ricin cytotoxicity. Somatic cell hybridization between the Vero and BER-40 cells showed that the BFA-resistance in BER-40 behaved as a codominant trait. The structure of the Golgi region, as examined by immunofluorescence microscopy with antibodies against Golgi markers (the 110-kDa protein and mannosidase II) or with fluorescent lipid NBD-ceramide, was unchanged in the mutant cells as compared to that in the wild-type cells. Treatment of Vero cells with BFA (1 micrograms/ml) or with 2-deoxyglucose plus sodium azide resulted in a rapid release of the 110-kDa protein, mannosidase II, and NBD-ceramide from the Golgi membrane to a more diffuse distribution in the cytosol. In contrast, these three Golgi markers remained to be Golgi-associated following treatment of BER-40 cells with BFA or with 2-deoxyglucose plus sodium azide. Immunoblotting of cell extracts from Vero and BER-40 cells with monoclonal antibody against the 110-kDa protein did not reveal any significant difference in the level of this Golgi marker in the mutant cells. These data suggest that the BFA-resistance mutation in BER-40 has rendered the cyclic pathway of the 110-kDa protein assembly to the Golgi membrane resistant to both BFA and 2-deoxyglucose plus sodium azide.

Adenosine Triphosphate↗

PubMed1992

Export of the outer membrane lipoprotein is defective in secD, secE, and secF mutants of Escherichia coli.

The export of major outer membrane lipoprotein has been found to be affected in secD, secE, and secF mutants of Escherichia coli, which are defective in protein export in general. After a shift to the nonpermissive temperature, the kinetics of accumulation of prolipoprotein and pre-OmpA protein was indistinguishable from that of pre-OmpA protein accumulation in the secD and secF mutants but different in the secE mutant. The prolipoprotein accumulated in the secD, secE, and secF mutants at the nonpermissive temperature was not modified with glyceride. We conclude from these results and those of previous studies that the export of lipoprotein requires all common sec gene products except the SecB protein, i.e., the SecA, SecD, SecE, SecF, and SecY proteins.

Bacterial Outer Membrane Proteins↗

PubMed1992

Use of indomethacin in the prophylaxis of ureteral colic following extracorporeal shock wave lithotripsy.

Ureteral obstruction leading to pain seems to be related to an increase of renal prostaglandins (PG). We designed a prospective double-blind, placebo controlled protocol for evaluating the effect of indomethacin, a PG-synthetase inhibitor, in the prophylaxis of ureteral colic following extracorporeal shock wave lithotripsy (ESWL). Sixty patients undergoing ESWL were randomized into two groups, group 1 (study group) received 50 mg indomethacin capsule three times daily and group 2 (control group) received multiple-vitamin tablet three times daily. Twenty-four hours urine samples were collected before and three days after ESWL was performed for PGE2 (predominant renal PG) determination. Subjective pain after ESWL was recorded with pain score (0-10 degrees). Oral codein or pethidine injection were available for relieving pain following ESWL. The pain score, analgesic requirement and urinary PGE2 in pre- and post-ESWL were used to compare the two groups. The pain score was 4.00 +/- 0.25 and 3.00 +/- 0.25 in the control and study groups respectively; it had a statistically significant difference (p < 0.01). In control group, 12 and 14 patients required 23 doses of codein and 18 doses of pethidine. In the study group, three and four patients required five doses of codein and eight doses of pethidine. The difference was statistically significant (p < 0.05). In the control group, the mean pre- and post-ESWL urinary PGE2 was 305 +/- 65.8 and 474 +/- 101 micrograms/24-hr respectively. In the study group, the mean pre- and post-ESWL urinary PGE2 was 289 +/- 60.7 and 186 +/- 26.5 micrograms/24-hr respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1992

[A comparison of transrectal fine needle aspiration and biopsy of prostatic lesion].

Transrectal fine needle aspiration and biopsy were performed on 100 patients with suspicious prostatic lesions using 22 gauge aspiration needles and 18 gauge biopsy needles. These collections were made from January 1990 to June 1991. Sufficient prostate tissue for cytological and histological diagnosis were obtained in 97% (97/100) and 99% (99/100), respectively. Thirty-nine patients (39%) were diagnosed having cancer with an accuracy rate of 89.7% (35/39) by histology and 69.2% (27/39) by cytology. One patient with a normal cytological aspiration was found to have an atypical biopsy, therefore, the biopsy was repeated and the prostate showed adenocarcinoma. Our results indicate that the needle aspiration cytology has a less accurate diagnosis rate than the needle biopsy histology. Fine needle aspiration is a safe and effective outpatient procedure but there is a definite learning curve before its use can be fully exploited. It is important to have experienced urologists and pathologists to perform these aspirations until sufficient experience is accumulated.

Aged↗

PubMed1992

Identification of beta-subunit of GTP-binding regulatory protein in mitotic spindle.

BACKGROUND: The heterotrimeric GTP-binding regulatory protein (G protein) is important in membrane signal transduction. Since the function of the beta-subunit of G protein (G beta) in tumor cells is not well-documented, identification of G beta in tumor cells was performed. EXPERIMENTAL DESIGN: Immunolocalization, Western blotting, and immunoprecipitation of G beta in mammalian tumor and normal cells were investigated using rabbit antisera against amino and carboxyl terminal peptide fragments of G beta. A human nasopharyngeal carcinoma cell line (NPC-TW039) was used as the cell model because of its short doubling time. RESULTS: Anti-G beta immunoreactivity was found to be associated with the plasma membrane and the mitotic spindle throughout the mitotic phase of cell replication. Colcemid pretreatment resulted in random distribution of the anti-G beta reaction product in the mitotic cells. The same phenomenon was also seen in various other tumor and normal cell lines. When solubilized membranous, cytosolic, and mitotic spindle fractions were analyzed by Western blotting, G beta (35 +/- 1 kDa) was found to be associated with the plasma membrane and mitotic spindle fractions. Immunoprecipitation of isolated mitotic spindles with anti-G beta further confirmed these results. CONCLUSIONS: These findings indicate that G beta is closely associated with the mitotic spindle as well as the plasma membrane and may be important in regulation of cell mitosis in addition to transmembrane signal transduction.

Amino Acid Sequence↗

PubMed1992

[Morphologic and biochemical characteristics of viridans streptococci isolated from dental plaque].

The purposes of this study were to give the characteristics of viridans streptococci isolated from dental plaque, and the relationships with dental caries. We stained 69 patients' teeth on supragingival plaques (Plaque Index = 1) with the Pfiffer's reagent or FD&C RED#28 dye, then isolated 80 strains of oral streptococci from 69 patients. Cultures were grown in TSB, MS, and MSB media. A total of 80 fresh-isolated plaque streptococci, including strains of S. salivarius, S. sanguis, S. mitis, and S. mutans were identified on the basis of morphologic, dark field, SEM, and biochemical characteristics. All of the oral plaque streptococci were to be found in significant percentages in dental plaque. Patients who had caries harboured S. mutans more often (37/40) than patients without caries (7/29). S. mutans comprised the greatest percentage (37/69) of microorganisms encountered in caries' plaque, and there was an association between high counts of S. mutans and the presence of dental caries. Cultural and biochemical methods were used for the isolation and identification of the different species of S. mutans. The result indicated that biotype I were the most predominant species (50%) in dental plaque. Thus, it is possible to isolate, screen and identify unknown strains through the morphologic, darkfield, SEM, and biochemical characteristics of viridans streptococci isolated from human dental plaque. By means of the present methods for identification, we hold considerable promises for contributing to the prevention and diagnosis of dental diseases.

Adolescent↗

PubMed1991

Membrane topology of Escherichia coli prolipoprotein signal peptidase (signal peptidase II).

The lsp gene of Escherichia coli encodes the inner membrane enzyme, signal peptidase II (SPase II). SPase II is comprised of 164 amino acid residues and contains four hydrophobic domains. A series of lsp-phoA and lsp-lacZ gene fusions have been constructed in vitro to determine the topology of SPase II. The fusion junction for each of these gene fusions was determined by DNA sequencing. The lengths of the SPase II fragment in the fusions varied from 12 to 159 amino acid residues. Strains containing SPase II-PhoA fusions to the two predicted periplasmic loops exhibited higher levels of alkaline phosphatase activity than fusions to the predicted cytoplasmic domains. In contrast, SPase II-LacZ fusions at the cytoplasmic and the periplasmic domains of SPase II showed high and low levels of beta-galactosidase activity, respectively, a result opposite to those shown by SPase II-PhoA fusions located at precisely the same amino acid of SPase II. Taken together, these results strongly support the predicted model for SPase II topology, i.e. this enzyme spans the cytoplasmic membrane four times with both the amino and the carboxyl termini facing the cytoplasm.

Amino Acid Sequence↗