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Biomedical subjects

F Wong

Publications and source records attributed to F Wong.

17 recordsLinked to original sources

A novel regulatory myosin light chain gene distinguishes pre-B cell subsets and is IL-7 inducible.

We describe a novel regulatory myosin light chain gene (termed precursor lymphocyte-specific regulatory light chain or PLRLC) that is expressed specifically in precursor B and T lymphocytes. PLRLC is the first example of a regulatory myosin light chain gene which displays specific expression in non-muscle cells. PLRLC is expressed in adult bone marrow derived normal and transformed pre-B cells; in the former, PLRLC expression levels are induced by the pre-B cell specific growth factor interleukin-7 (IL-7). PLRLC is not expressed in either transformed pre-B cells derived from fetal liver or in normal fetal liver pre-B clones grown in the presence of IL-7. Therefore this gene provides the first marker that clearly distinguishes these two pre-B subsets. Finally, several of the different PLRLC transcripts potentially encode regulatory myosin light chains with unique structural features. The unique distribution, regulation and structural features of the PLRLC gene products suggest an important role for PLRLC during lymphocyte development.

Amino Acid Sequence

Ascorbic acid modulates collagen type I gene expression by cells from an eye tissue--trabecular meshwork.

The trabecular meshwork, a specialized tissue in the anterior chamber of the eye, plays a major role in the regulation of aqueous humor outflow. We studied the effects of ascorbic acid, a significant component in the aqueous humor, on gene expression of type I collagen in cultures of bovine trabecular meshwork cells. These cells were plated for 6 days, exposed to ascorbic acid in concentrations of 100, 250 and 500 micrograms/ml for 3 days and labeled with (3H)proline for the last 24 hrs. Cultures that did not receive ascorbic acid served as controls. Bacterial collagenase assays showed enhanced incorporation of (3H)proline into collagenous proteins in cultures treated with 100 and 250 micrograms/ml of ascorbic acid. Gel electrophoresis and fluorography revealed that ascorbic acid caused a 2.6- to 4.9-fold increase in production of alpha 1 (I) and alpha 2(I) collagen chains by trabecular meshwork cells. Such an increase was found, using a cDNA probe specific for pro alpha 1(I) chains, to be accompanied by an increase in steady-state mRNA levels. Similar findings were also yielded from in situ hybridization experiments. These results, coupled with previously demonstrated ascorbate-induced effects on glycosaminoglycan, fibronectin and laminin synthesis, suggest that ascorbic acid is a key mediator of the extracellular matrix production by trabecular meshwork cells. Fluctuations in its concentration may lead to alterations in the makeup and assembly of matrices underlying the cells.

Animals

Hygromycin B therapy of a murine coronaviral hepatitis.

Hepatitis caused by mouse hepatitis virus (MHV-A59), a murine coronavirus, is accompanied by direct infection and replication of virus within the liver. We demonstrate here that the aminoglycoside hygromycin B is able to eliminate MHV-A59 infection from mouse peritoneal macrophages and cultured liver cells in vitro and is also able to reduce levels of virus replication and necrotic liver foci in vivo.

Animals

On the membrane cytopathology of mouse hepatitis virus infection as probed by a semi-permeable translation-inhibiting drug.

Previous studies of the membrane fusion process have permitted the characterization of membrane permeability changes concomitant with MHV-induced cytopathology. One indication of membrane permeability in MHV-infected cells is their sensitivity to translation inhibition by the normally impermeable amino-glycoside, hygromycin B (Macintyre, G., Wong, F. and Anderson, R. (1989) J. Gen. Virol. 70, 763-768). In the present study, we examine the hygromycin B sensitivity of acutely infected mouse fibroblast L-2 cell and macrophage cultures as well as persistently infected mouse fibroblast LM-K cell cultures. The results suggest that membrane permeability alterations (as indicated by hygromycin B sensitivity) are a common feature of these MHV infections. Hygromycin B "cured" persistently infected LM-K cells as indicated by the absence of detectable virus antigen by immunofluorescence and by the absence of infectious virus even after removal of the drug or co-cultivation with untreated L-2 cells. The results argue against the maintenance of MHV infection by a mechanism involving latently or non-cytolytically infected cells. We conclude therefore that at least one mechanism for MHV persistence depends on virus propagation by cytolytic infection of a small, dynamically changing, fraction of the total cells present in culture.

Animals

Mouse fibroblast mutants selected for survival against mouse hepatitis virus infection show increased resistance to infection and virus-induced cell fusion.

We describe here a genetic approach to the analysis of host cell functions involved in determining permissiveness to mouse hepatitis virus (MHV). Using the chemical mutagen, ethyl methane sulfonate (EMS), mouse fibroblast cell mutants were generated which were selected for resistance to cell-killing by MHV. These mutants were then screened for their susceptibility to MHV infection, ability to replicate MHV and relative sensitivity to MHV-induced cell fusion. In contrast to wild type L-2 cells which were acutely and terminally infected by MHV, all five mutants examined replicated MHV in a persistent manner. These mutants showed a reduced susceptibility to MHV infection and an increased resistance to MHV-induced cell fusion. Fusion resistance was specific to that mediated by the MHV E2 protein; mutant as well as wild type L-2 cells were equally sensitive to fusion by polyethylene glycol. The combined effect of reduced infectability and increased fusion resistance was to limit MHV infection to only a small percentage of the total cells in culture, thereby permitting survival of both virus and cells. The observed high rate of generation of the cell mutants suggests that the conversion of a fully MHV-susceptible cell to a semi-resistant one (capable of supporting a persistent infection) is a fairly common event, possibly involving a single mutation.

Animals

Progestational and androgenic receptor binding affinities and in vivo activities of norgestimate and other progestins.

The progestational and androgenic in vitro receptor binding affinity and the in vivo activity of norgestimate was compared with that of its metabolites and other progestins. The relative binding affinities (RBAs) of norgestimate and its 17-deacetylated metabolite for rabbit uterine progestin receptors were similar to that of progesterone (P); those of 3-keto norgestimate and levonorgestrel were about five times that of P; those of gestodene and 3-keto desogestrel were about nine times that of P. The RBAs of norgestimate, P, and 3-keto norgestimate for rat prostatic androgen receptors were from 0.003 to 0.025 times that of dihydrotestosterone (DHT); those of 3-keto desogestrel, gestodene, and levonorgestrel were from 0.118 to 0.220 times that of DHT. The order of receptor level selectivity represented by the ratio of androgen:progestin IC50 values (with a greater ratio value reflecting a better selectivity) was norgestimate greater than P = 3-keto norgestimate greater than 17-deacetylated norgestimate greater than 3-keto desogestrel greater than gestodene greater than levonorgestrel. In vivo studies demonstrated similar profiles for norgestimate and its 17-deacetylated metabolite. These latter two steroids were equally potent as progestins in stimulating rabbit endometrium, and compared with the other progestins, both steroids exhibited minimal androgenicity as measured by the stimulation of rat prostate growth. In conclusion, these studies, as well as previous preclinical and clinical studies, provide evidence of the selectivity of norgestimate based on minimal androgenicity, indicating an improvement over other progestins used in oral contraceptives.

Animals

The usefulness of ultrasonography in intracavitary radiotherapy using selectron applicators.

Uterine perforation can occur during the procedure of intracavitary insertions for the treatment of carcinoma of the cervix. If radiation treatment proceeds in the situation of inadvertent perforation, severe complication may result. The ultrasonographic studies of five patients with metal intrauterine applicators in situ are described. The applicator appeared as a linear echogenic density within the uterine cavity. The thickness of the uterine wall and the fundus could be clearly identified in a normal insertion. In the presence of uterine perforation, the applicator was seen traversing the myometrium necessitating alteration of the treatment plan.

Brachytherapy

An immunopathologic study of retinoblastoma protein.

We investigated the immunoreactivity of the retinoblastoma protein in eight retinoblastoma tumors of different stages of differentiation. All undifferentiated tumor cells failed to show immunoreactivity. In one patient, who had a family history of retinoblastoma and had been treated by radiation, the tumor had a large area of undifferentiated cells and small independent retinocytomas. The tumor cells in the retinocytoma and the cone cells in the adjacent retina, but not the undifferentiated tumor cells, showed positive staining with two anti-Rb antibodies. The immunoreactivity may be related to radiation or differentiation of retinoblastoma and cone cells.

Child, Preschool

Frequency characteristics in the visual system of Drosophila: genetic dissection of electroretinogram components.

Various drosophila mutants were used to dissect the electroretinogram (ERG) frequency response into components of different origins. The ommochrome granules in the receptor cell body are known to migrate in response to light, limiting the amount of light entering the rhabdomere. Comparison between the ERG frequency responses of the wild type and the mutant lacking the ommochrome granules indicates that the pigment migration reduces the amplitude gain at frequencies below 0.5 Hz. The ERG of drosophila compound eyes consists of contributions from receptor cells and the second-order cells in the lamina. Mutants with defective laminae showed a high-frequency cutoff with a corner frequency of about 20 Hz, while in wild type the response peaked in that frequency region. These results suggest that the lamina contributes mainly to the high-frequency components of the ERG transfer function. The shot noise model (Dodge et al., 1968) has been tested in drosophila by comparing the frequency response of the superimposed on the intracellular receptor potential. The results are consistent with the hypothesis that the receptor potential consists of a summation of small discrete potentials (bumps). In a mutant in which the bumps exhibit latency dispersion in response to a dim flash, the receptor showed a poor high-frequency response, the corner frequency being lowered to about 1-2 Hz. The slope of the cutoff was approximately 20 dB/dec indicating that the latency dispersion in this mutant is the major limiting factor in temporal resolution. Light-evoked high frequency oscillations have been observed in the ERG of another mutant. The oscillation was found sharply turned to light flickering at about 55 Hz.

Animals

Lability of the prolonged depolarizing afterpotential in Balanus photoreceptors.

A large cell to cell variability of the prolonged depolarizing afterpotential (PDA) decay time constant (tau) has been measured in Balanus eberneus lateral ocelli. While 25% of the cells had PDA's of long duration, tau greater than 10 min. 45% of the cells tested showed either weak (tau less than 60 s) PDA or none at all. The variability was not reflected in the late receptor potential. All the cells showed normal light-coincident responses. The variability was not due to some alteration of the thermal stability of the pigment states, since after monochromatic adaptation the amplitude of the early receptor potential remained unchanged for at least 30 min. In addition, in some cells that initially showed PDAs of long duration, the decay time was either shortened or abolished after exposure to anoxia. Again, the late receptor potential and the stability of the pigments remained unaffected. These results indicate that the mechanisms which give rise to the PDA are not always tightly coupled to the direct chain of events that lead to the light-coincident response.

Action Potentials

Phase I study of concurrent carboplatin and radiotherapy in previously untreated patients with stage III and IV head and neck cancer.

This study evaluated the toxicity resulting from combined therapy using carboplatin and radiotherapy in 22 patients with locally advanced (stage III and IV) head and neck cancer. Carboplatin was given after radiotherapy during the first 5 consecutive days of a 16-fraction course of radiotherapy delivered in a total of 22 days. To find the acceptable toxic dose of carboplatin, the patients were treated in groups of 3 patients each. The first group received 50 mg/m2 carboplatin and the dose was increased by 10 mg/m2 in each subsequent group. Unacceptable toxicity (severe mucositis, nausea and vomiting, and/or myelosuppression) was encountered at a carboplatin dose of 80 mg/m2. A further 10 patients were then treated at a dose of 70 mg/m2. At this dose toxicity was acceptable. Although the group of 22 patients is too small for response and survival data to be meaningful, 10 complete responses were seen and median survival is in excess of 67 weeks.

Aged

Presence of p53 mutation in human cervical carcinomas associated with HPV-33 infection.

In this present study, we report the mutation of the p53 gene in vivo in human primary carcinomas of cervix and cervical intraepithelial neoplasia (CIN). The association of the HPV subtypes with the tumors was determined by multiplex primer polymerase chain reaction (PCR) amplification. The mutation of the p53 gene was detected using PCR amplification of the p53 exons followed by SSCP (single strand conformation polymorphism) and DNA sequencing analysis. The p53 mutation was detected in two out of two HPV-33 positive carcinomas but was absent in the HPV-16/-18 positive carcinomas (0 out of 8 cases). The p53 mutation was also detected in one out of four HPV-negative cervical carcinomas. No mutation of the p53 gene was detected in the CIN specimens (0 out of 7 cases). The two mutations in the HPV-33 associated cervical carcinoma were detected at codon 273 (CGT to TGT; arginine to cysteine) and intron 5 (24 base pair downstream of the 3' end of exon 5). The p53 mutation at codon 273 has been previously reported in one of the HPV-negative cervical carcinoma cell line (C33A). Our results indicate that mutation of the p53 gene is not a common event in human cervical cancers (3/14), and may be related to the infection of HPV-16/18 in the tumor. However, mutation of the p53 gene was detected in cervical carcinomas associated with HPV-33 and may be an important genetic event in this subgroup of carcinomas.

Amino Acid Sequence