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Biomedical subjects

M J Wilson

Publications and source records attributed to M J Wilson.

At least 127 records · Page 7Linked to original sources
PubMed1989

Localization of cathepsin B in normal and hyperplastic human prostate by immunoperoxidase and protein A-gold techniques.

Cathepsin B, a lysosomal cysteine protease, was localized in normal prostate and benign prostatic hyperplasia (BPH) using immunoperoxidase and protein A-gold techniques. Our objective was to determine whether cathepsin B was involved in the prostatic epithelium affected by nodular hyperplasia. All samples were collected immediately after prostatectomy. Immunohistochemical studies showed that the enzyme was expressed in the supranuclear cytoplasm of columnar cells and in numerous basal cells of normal and BPH acini. The strongest localization of cathepsin B occurred in acinar basal cells; hence, it is possible that cathepsin B could be useful as a marker for such cellular elements. Stromal macrophages showed reaction products, but lymphocytes and neutrophils did not. In both normal and hyperplastic glands, the enzyme was localized by gold particles in lysosomes, secretory granules, and vacuoles of columnar epithelial acinar cells. Immunoelectron microscopic study also showed the presence of cathepsin B in the heterochromatin (condensed chromatin) and nuclear membranes of columnar and basal cells, but not in euchromatin or nucleoli. At present, the function of cathepsin B in the nuclei of basal and columnar cells remains unknown. However, the cathepsin B in the cytoplasmic compartment might be associated with the lysosomal function of the cells. The role of cathepsin B as a marker for basal cell participation in the development of prostatic lesions should be studied further.

Cathepsin B↗
PubMed1989

Immunohistochemical localization of laminin in the basement membranes of normal, hyperplastic, and neoplastic human prostate.

We examined the effects of fixatives and antibody sources on the immunohistologic localization of laminin in normal and cancer-containing human prostates and studied the localization patterns in carcinomas of varying degrees of histologic differentiation. Two different polyclonal antibodies were localized in paraffin-embedded or cryostat sections of fixed (alcohol, formalin, and paraformaldehyde) or unfixed tissue, using the immunofluorescence (IF) or immunoperoxidase (IP) techniques, with positive and negative controls. We found that the IF reactions were more intense in unfixed or alcohol-fixed sections than in paraformaldehyde-fixed specimens. IP reactions were very weak or absent in fixed and paraffin-embedded sections, but pepsin treatment of these sections resulted in more intense and uniform IP reaction products, stronger than in unfixed or ethanol-fixed cryostat sections. With the IP technique, laminin localization was intense and uniform in the basement membranes (BM) of acini, blood vessels, smooth muscle, and nerve fibers in normal prostate, benign hyperplasia (BPH), and well-differentiated carcinomas. The BM of poorly differentiated carcinomas showed widespread absence of laminin reactivity. In normal BPH and well-differentiated tumors, occasional epithelial cells and their surface and acinar lumina had laminin reactivity. However, in higher grade tumors, numerous neoplastic cells had laminin reactivity in cytoplasm, their surface, and secretory material. Some macrophages and neutrophils also contained laminin reactivity, presumably of degraded laminin. In some moderately and poorly differentiated tumors, the BM of small capillaries did not contain laminin. The BM of larger vessels always had laminin reactivity, even in the higher grade tumors.

Adenocarcinoma↗
PubMed1989

Plasminogen activators in human prostate cancer cell lines and tumors: correlation with the aggressive phenotype.

The levels of several tumor associated proteases, including plasminogen activators (PA), are elevated in many malignant tumors compared to their benign tumor counterparts. Extracellular matrix degradation mediated by PA may facilitate tumor cell invasion and metastasis. To assess whether PA content correlates with the aggressive phenotype in prostate cancer, we studied these activators in the PC-3 human prostate cell line and PC-3CALN, an aggressive in vivo derived variant cell line. Enzymatic assays using H-D-val-leu-lys-pNA (S-2251) as substrate and peroxidase-anti-peroxidase immunohistochemical techniques were used. In an in vitro chemoinvasion assay, the PC-3CALN variant cell line demonstrated significantly greater invasive behavior than the unselected, parental PC-3 line. The activity of PA secreted by PC-3CALN cells was 3.5 times greater than that of PC-3 cells (p less than 0.01). PC-3 metastases obtained following intrasplenic injection of PC-3 cells had greater PA activities than the corresponding primary tumors. Immunohistochemical studies of PC-3 tumors demonstrated preferential localization of urokinase-type PA to areas of apparent tumor cell invasion. These data suggest a correlation between PA and the aggressive phenotype in this model of human prostate cancer. PA, in particular u-PA, may play a role in the migration and invasion of prostate cancer cells and provide a marker of the aggressive phenotype.

Animals↗
PubMed1989

Analysis of auditory evoked potentials by magnitude-squared coherence.

Evoked potentials are usually analyzed in the time domain (voltage versus time). The most familiar frequency-domain measure, the power spectral density function, displays power as a function of frequency but doesn't distinguish signal power from noise power. The coherence function estimates, for each frequency, the ratio of signal power to total (signal plus noise) power and, thus, indicates the degree to which system output (scalp potential) is determined by input (acoustic stimulus). Coherence ranges from 0 to 1; values above specified critical values can be accepted as demonstrating statistically significant system response. In this paper, we present coherence analysis of human scalp responses to clicks and amplitude-modulated tones. In both cases, this analytic method provides insight into the spectral character of the response (for example, assisting in specifying desirable filter characteristics). Threshold sensitivity is also improved: statistically significant responses can be detected at lower intensity by coherence analysis than by inspection of time-domain waveforms.

Adult↗
PubMed1989

Electrophysiologic assessment of low-frequency hearing: sedation effects.

Sixteen normal-hearing adults were tested in both awake and sedated states for several evoked responses to low-frequency stimuli. Responses obtained by cross-correlation function, middle latency responses, and the 40-Hz response proved most sensitive; all had mean thresholds of less than 20 dB normal hearing level for the awake-alert state, but 40-Hz and middle latency response mean thresholds were shifted about 10 dB under sedation. The cross-correlation method seems to offer promise for pediatric auditory assessment.

Adult↗
PubMed1988

Relationship of prostatic acid phosphatase localization in human prostate by a monoclonal antibody with the Gleason grading system.

Prostatic acid phosphatase (PAP) was localized in human prostate with a monoclonal antibody prepared against PAP isoenzyme II to determine patterns of its expression in normal, hyperplastic (BPH), and cancerous glands. The monoclonal antibody reacted with both isoenzymes II and IV in immunoblot studies. Formalin-fixed, paraffin-embedded tissue was used from patients who had not been treated with hormones or chemotherapy. In normal glands and BPH, there was marked variation in the intensity of PAP staining in morphologically otherwise similar epithelial cells. There was similar heterogeneity of staining in the adenocarcinomas. Rough quantification of the intensity patterns in the clinical groups indicated a slight shift to more intense staining in BPH and well-differentiated carcinomas but a progressive decline in the PAP staining in the moderately and poorly differentiated tumors. This decrease in intracellular staining with decreasing differentiation is not inconsistent with the clinical observation that serum levels of acid phosphatase generally increase with higher grade and disseminated tumors, since the enzyme is simply more accessible to the circulatory system in those cases. The same decrease may explain the few disseminated tumors that are not associated with elevated serum levels.

Acid Phosphatase↗
PubMed1988

Increased metallothionein gene expression in 5-aza-2'-deoxycytidine-induced resistance to cadmium cytotoxicity.

The pyrimidine analog, 5-azacytidine (AZA-CR), has been shown to increase the expression of the metallothionein (MT) gene and to induce tolerance to cadmium toxicity. Since incorporation into DNA of AZA-CR appears to be required for this effect, the deoxynucleoside of AZA-CR should also be effective. Therefore, this study was undertaken to assess the effect of 5-aza-2'-deoxycytidine (AZA-CdR) pretreatment on cadmium-induced cytotoxicity and MT expression in cultured cells. TRL 1215 cells in log phase of growth were exposed to AZA-CdR (0.4, 0.8, 4.0, 8.0 microM) followed 48 h later by the addition of cadmium (10 microM). MT concentrations were measured 24 h after the addition of cadmium. AZA-CdR alone caused modest, dose-related increases in MT levels (2.3-fold maximum), while cadmium alone resulted in a 9.5-fold increase. Pretreatment with AZA-CdR in combination with cadmium caused a 19--24-fold increase in cellular MT at all doses of AZA-CdR. Addition of the DNA synthesis inhibitor, hydroxyurea (HU), to the incubation medium during AZA-CdR exposure prevented the enhancing effect of the analog on cadmium induction of MT accumulation. Time course studies revealed that AZA-CdR pretreatment reduced the time required for cadmium to induce MT levels from 4--8 h to 0--2 h. AZA-CdR pretreated cells placed in suspension with cadmium (125 microM) showed a marked reduction in cadmium-induced cytotoxicity as reflected by reduced glutamic-oxaloacetic transaminase (GOT) loss. Uptake studies showed that AZA-CdR pretreatment had no effect on cadmium transport during the initial phases of exposure, indicating that an alteration in the toxicokinetics of the metal did not account for the reduction in toxicity. AZA-CdR did, however, cause hypomethylation of the MT-I gene. These results suggest that AZA-CdR pretreatment induces tolerance to cadmium toxicity by increasing the genetic expression of MT possibly through hypomethylation of the MT gene.

Animals↗
PubMed1988

The Ca2+-dependent protease inhibitor of rat ventral prostate: properties of the inhibitor and effects of castration on Ca2+-dependent protease and inhibitor activities.

1. The rat ventral prostate contains a heat stable inhibitor of Ca2+-dependent protease. This inhibitor was found to exist in a wide range of molecular weights (approx. 40-270 kDa) in adult rats. 2. However, in rats immediately post puberty (45 days of age) the inhibitor was predominantly of the higher molecular weight forms. 3. The inhibitor was also found in the dorsolateral and anterior (coagulating gland) prostate lobes but was of lower specific activity than in the ventral lobe. 4. Although the activities of the Ca2+-dependent protease and inhibitor decreased per ventral prostate gland after castration, these activities were not different during the first 10 days postcastration when expressed per g wet wt or per unit cytosol protein. 5. With a longer duration of castration, there was a decline in the specific activity (per unit protein) of the protease and an increase in that of the inhibitor. 6. Thus, the activities of the protease and inhibitor change in concert with the amount of cellular cytosol protein during the active period of castration-induced atrophy. 7. However, in long term castrated rats, functions carried out by the Ca2+-dependent protease may be effectively suppressed. 8. These data suggest that the Ca2+-activated protease probably is involved in the regulation of some metabolic processes in the active gland and is not prominent in the castration induced atrophy of the ventral prostate unless it functions through the proteolysis of some select protein(s).

Animals↗
PubMed1988

Methylation status and organization of the metallothionein-I gene in livers and testes of strains of mice resistant and susceptible to cadmium.

The methylation status, copy number and organization of the metallothionein-I (MT-I) gene was studied in hepatic and testicular DNAs of mouse strains resistant (BALB/c) and susceptible (NFS) to cadmium-induced testicular toxicity. Digestion of DNAs by the restriction enzymes BamHI, EcoRI and HindIII produced identical patterns for hepatic and testicular DNAs of both strains, indicating that there was no apparent difference in the gross genomic organization or in copy number of the MT-I gene in the 2 types of tissues from either strain. Digestion with MspI, HpaII, AvaII and HhaI indicated that the hepatic DNAs of both strains were under-methylated as compared to the testicular DNAs. However, the NFS DNAs lacked a fragment that was consistently observed in the MspI digests of BALB/c DNAs, suggesting the presence of a polymorphic CCGG site. This site was localized by double digestion of DNAs with BstEII or HindIII and MspI to the 3' end of the MT-I gene. Differences in methylation status may account for the differential susceptibility of the 2 tissues to cadmium toxicity. The higher degree of MT-I gene methylation may result in slower or inefficient induction of MT in the testes, resulting in greater sensitivity to metal toxicity in testes than in liver. However, differences in methylation status alone do not seem to account for the interstrain differences in cadmium toxicity, and other factors, such as differences in genetic organization, seem to be involved in the inducibility of MT-I gene in different strains.

Animals↗
PubMed1988

Auditory responses to the envelopes of pseudorandom noise stimuli in humans.

Averaged scalp potentials evoked by continuous pseudorandom noise can be cross-correlated with the evoking stimulus, yielding a cross-correlation function (CCF) which reflects neural phase-locking and is quite sensitive for low-frequency stimulus components [M.J. Wilson and R.A. Dobie (1987) Electroencephalogr. Clin. Neurophysiol. 66, 529-538]. However, for higher frequency signals, replicable CCFs can only be obtained at moderate to high intensities. Since auditory neurons also respond to envelopes of complex sounds, even for high-frequency carriers, we compared scalp responses evoked by band-limited complex sounds to the envelopes of these sounds; the resultant envelope cross-correlation functions (ECCFs) contained replicable response components primarily below 1,000 Hz, regardless of the evoking stimulus spectrum. ECCF thresholds for three octave-band stimuli (830-1,562, 1,611-3,125, and 3,174-6,201 Hz) were more sensitive than CCF thresholds (P = 0.006), averaging 35 dB spectrum level for 10 normal subjects. When stimuli with only odd harmonics were used, replicable odd-component scalp responses were seen only in the spectral range of the stimuli, while even-component responses (presumably to stimulus envelope) were seen only in low-passed scalp responses.

Acoustic Stimulation↗
PubMed1988

c-H-ras and c-K-ras gene hypomethylation in the livers and hepatomas of rats fed methyl-deficient, amino acid-defined diets.

The extent of methylation of the c-H-ras and c-K-ras oncogenes was compared in neoplastic and preneoplastic livers of rats fed one of several methyl-deficient, amino acid-defined diets for 18 months, with or without a preceding initiating dose of diethylnitrosamine (DEN). The restriction endonucleases MspI, HpaII, HhaI, PaeR71 and XhoI were used for studying the extent and pattern of DNA methylation. The results indicated that both c-H-ras and c-K-ras oncogenes were hypomethylated in all DNA samples derived from both neoplastic and preneoplastic livers of rats fed any of the methyl-deficient diets used, regardless of whether or not the rats had received an initiating dose of DEN. It thus appears that dietary methyl deficiency does indeed lead to hypomethylation of ras genes in the DNAs of the resulting tumors. However, the significance of this hypomethylation in the tumorigenic process is not clearly understood.

Amino Acids↗
PubMed1988

An epidemiologic study of craniosynostosis: risk indicators for the occurrence of craniosynostosis in Colorado.

This population-based case-control study was designed to investigate risk indicators for the occurrence of the birth defect craniosynostosis in Colorado. A total of 173 children who underwent craniectomy for craniosynostosis and 759 children without craniosynostosis were included in the study. Multivariable logistic regression analysis of birth certificate data showed that male sex (odds ratio (OR) = 1.6, 95% confidence interval (CI) = 1.1-2.2), maternal five-year age "increase" (OR = 1.3, 95% CI = 1.1-1.5), plural birth (OR = 3.0, 95% CI = 1.2-7.1) and black maternal race (OR = 0.0, 95 per cent CI = 0.0-0.6) were independently associated with craniosynostosis. There was a weak positive association between craniosynostosis and the altitude of the town closest to the maternal residence, but no association was found with maternal education, marital status, number of previous births, or previous pregnancy termination. The association of craniosynostosis with plural birth is consistent with the hypothesis of fetal head constraint.

Age Factors↗
PubMed1988

Plasminogen activator activities in the developing rat prostate.

Plasminogen activator (PA) activities were measured in the rat prostatic complex and individual prostatic lobes during early postnatal and pubertal development and in sexually mature adult rats. There was no significant change in PA activity during postnatal prostate development. However, during sexual maturation with puberty, there was a decline in PA activity in the ventral (3-fold), dorsolateral (22-fold), and anterior (19-fold) prostate lobes when activity was expressed per unit protein. A decrease in activity of 25- and 11-fold was found for the dorsolateral and anterior lobes, respectively, when activity was expressed per unit DNA. There was no change in activity in the ventral lobe. The adult ventral prostate (and its secretion) have 3 broad bands of low molecular mass (approximately 23 and 26-32 kDa) plasminogen-independent protease activities. Proteases of these molecular sizes as well as an activity of 170 kDa were detected in the dorsolateral prostate. The former proteases in the ventral and dorsolateral lobes were first found at 21 days of age, whereas the 170 kDa protease was found in dorsolateral prostate immediately post-puberty (48 days). The low molecular mass plasminogen-independent proteases were also able to activate plasminogen (determined by zymography) and hence contribute to the total measured PA activity. Thus, at 21 days of age, the specific activity of plasminogen-dependent protease declined, since the total measured PA-specific activity did not change. Plasminogen-dependent activities in ventral, dorsolateral, and anterior prostate lobes of adult rats were found as doublets of approximately 57-59 kDa and 36-38 kDa.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Membrane-associated protein kinase activities in seminal plasma from vasectomized men.

Differential centrifugation was used to prepare heavy and light membrane fractions from the seminal plasma of vasectomized men. The two membrane fractions combined contained half of the phosvitin and histone kinase activities but only 7% of the total protein content in vasectomy semen. These two kinase activities as well as phosphorylation of endogenous membrane proteins were optimally stimulated by Mg2+; Mn2+ could effectively substitute for Mg2+ only in endogenous phosphorylation reactions. Neither the phosvitin nor histone kinase responded to cAMP or cGMP, but the histone kinase was strongly inhibited by the heat-stable cAMP-dependent protein kinase inhibitor. The phosvitin kinase was not affected by this inhibitor. The phosphorylation of endogenous proteins in the heavy membrane fraction was not affected by the protein kinase inhibitor but protein phosphorylation in the light membrane fraction was partly (45%) inhibited. The differential effects of increased ionic strength, sulphydryl protecting agents, and the protein kinase inhibitor on protein kinase activity towards lysine-rich histones, phosvitin and endogenous proteins, as well as differential extractability and binding to an anion exchange column of histone kinase and phosvitin kinase activities, indicate that more than one kinase activity is present in these membrane subfractions. Electron microscopic examination showed that there are several kinds of membrane-limited components in vasectomy seminal fluid that vary in size, density, and ultrastructure. The association of type(s) of protein kinase to individual membrane components remains to be established.

Cell Membrane↗
PubMed1987

Immunoelectron microscopic localization of prostatic-specific antigen in human prostate by the protein A-gold complex.

Human prostate specimens from 25 patients provided 15 normal, four benign prostatic hypertrophy (BPH), and seven adenocarcinoma samples which were studied using a polyclonal antibody against prostatic specific antigen (PSA) and protein A-gold complex. Our study showed the presence of gold particles in cytoplasmic vesicles and granules, rough endoplasmic reticulum, and occasional lysosomal dense bodies in columnar or cuboidal cells of acini from normal, BPH and well-differentiated cancerous specimens, but not in the acinar basal cells. Some moderately differentiated and most poorly differentiated tumors contained undifferentiated neoplastic cells in which PSA localizations often were associated with membranous structures since specific cytoplasmic organelles were not differentiated. In prostatic stroma, invasive cells which were of differentiated type localized gold particles. In addition, some neutrophils and macrophages also localized PSA suggesting their role in phagocytosis of extracellularly released PSA in the stroma of BPH and cancerous samples. Thus far, the authors have not observed neutrophils and macrophages with gold particles in normal prostate. We suggest that neutrophils and macrophages were involved in transport of some extracellularly released PSA from the prostatic stroma or other metastatic sites to sera in BPH and cancer patients.

Adenocarcinoma↗
PubMed1987

Studies of the Drosophila norpA phototransduction mutant. I. Electrophysiological changes and the offsetting effect of light.

The electrophysiological characteristics of norpAH52, a temperature sensitive phototransduction mutant of Drosophila melanogaster, were studied in vivo. Upon raising the environmental temperature to 33-37 degrees C, mutant flies exhibited time-dependent changes in photoresponses. Initial observations were losses in responsiveness at low light intensities and prolonged receptor potential waveforms. Next, reductions in response amplitudes at higher light intensities occurred, until no responses were obtained. On return to lower temperature the electrophysiological properties recovered in reverse order. Based on these observations we conclude that the primary defect of norpA affects the efficiency of the phototransduction process. Enhanced light exposure could offset the receptor potential changes in norpA. With the temperature sensitive mutant: (1) additional light exposure prolonged the time that responses could be observed at the higher temperature, (2) when 1-s illuminations no longer elicited responses at the higher temperature, 1-min illuminations at the same intensity temporarily restored the ability to obtain 1-s-responses, and (3) light accelerated the restoration of responses on return to lower temperature. Illumination also had an effect on non-temperature sensitive norpA mutants, enabling the production of small photoresponses in norpAH44, a mutant that normally does not exhibit any responses, and improving the low-light-intensity responses of norpAP16. Our study indicates that the PI cycle, which is inhibited in norpA mutants (Yoshioka et al. 1985), is an important light-sensitive positive step or effector in the production of receptor potential responses.

Animals↗
PubMed1987

Human short-latency auditory responses obtained by cross-correlation.

Short-latency auditory responses were obtained by cross-correlation of continuous, pseudorandom noise stimuli with averaged scalp potentials from adults with normal hearing. Responses were recorded for spectrum levels of 14-74 dB for noise bandwidths from 800 to 6000 Hz. At the lowest intensity level of broadband noise, all 10 subjects showed replicable cross-correlation functions (CCFs), which were characterized by prominent positive peaks at delays (latencies) of 5-7 msec. Male subjects exhibited longer delays than females. Delay (latency) increased with decreasing stimulus intensity. Very early responses (less than 2 msec) attributable to cochlear microphonic, which were prominent in earlier work on guinea pigs, were not well seen in these human data. CCFs for responses to band-limited stimuli and off-line derivation of band-limited CCFs for responses evoked by broadband stimuli both showed that this technique is most sensitive to frequency-following behavior at low frequencies (less than 800 Hz). However, definite phase-locked responses to even the highest passband (3100-6200 Hz) were seen. These results support the use of the CCF technique as an efficient method of frequency-specific assessment of the auditory system.

Adult↗