[Effect and mechanism of adipocyte co-culture on aquaporin-9 expression in HepG2 cells].

Objective: To observe the effect of differentiated mature adipocytes on hepatic steatosis and aquaporin-9 (AQP9) expressions in HepG2 cells and further explore its possible mechanism of action. Methods: Human preadipocytes were cultured and differentiated to full maturity. HepG2 cells were co-cultured with non-differentiated adipocytes and differentiated mature adipocytes for 48 h, and then labeled as control group and experimental group. Oil red O staining and intracellular triglyceride content were performed on co-cultured HepG2 cells and simultaneous changes in phosphatidylinositol 3-kinase (PI3K) - serine/threonine kinase (Akt) signaling pathway, and AQP9 mRNA and protein levels were detected. The experimental group was co-cultured with recombinant human insulin-like growth factor-I (IGF-I), with the addition of 100ng/ml PI3K-Akt pathway agonist, labeled as experimental group + IGF-I group. The activation of PI3K-Akt pathway was verified by Western blotting (WB). The expression of AQP9 was detected by RT-q PCR and WB. The recombinant lentivirus LV-AQP9 or empty-loaded virus LV-PWPI was transfected with HepG2 cells by recombinant lentiviral transfection tecnique, and labeled as HepG2-AQP9 and HepG2-PWPI. The transfection efficiency was assessed by confocal laser scanning microscopy and RT-qPCR and WB detected the change of AQP9 expression level after virus transfection. Afterwards, the stable over-expressed HepG2-AQP9 cells and the empty-loaded HepG2-PWPI cells were co-cultured with differentiated mature adipocytes for 48h, and labeled as HepG2-AQP9 co-culture group, and then intracellular triglyceride content were detected with Oil red O staining. Finally, IGF-I was added to the HepG2-AQP9 co-culture group, which was recorded as HepG2-AQP9 co-culture + IGF-I group. Intracellular triglyceride content was detected with Oil red O staining, and WB verified PI3K-Akt signaling pathway activation and changes in AQP9 mRNA and protein levels. A t-test was used to compare the two independent samples. Results: The intracellular lipid droplets and triglyceride content (0.052 ± 0.005) in the experimental group was increased significantly than the control group (0.033 ± 0.003) (t= 5.225,P= 0.006), suggesting that adipocyte co-culture had induced steatosis in HepG2 cells. RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (3.615 ± 0.330) and protein levels (0.072 ± 0.005) in the experimental group were significantly higher than the control group (t= 13.708, 11.225,P= 0.005, < 0.001). WB results showed that the expression level of phosphorylated Akt (p-Akt) protein (0.116±0.003) in the experimental group was significantly lower than the control group (0.202 ± 0.003) (t= 27.136,P< 0.001). The total Akt protein was constant, and the p-Akt/total Akt (0.182 ± 0.017)was significantly lower than the control group (0.327 ± 0.019) (t= 2.431,P= 0.001), suggesting that adipocyte co-culture had inhibited PI3K- Akt signaling pathway in HepG2 cells and up-regulated the expression level of AQP9. WB results indicated that the expression level of p-Akt protein (0.194 ± 0.021) in the experimental group + IGF-I group was significantly higher than the experimental group (0.132 ± 0.003) (t= 5.082,P= 0.007). The total Akt protein was constant, and the p-Akt/total Akt (0.281 ± 0.009) was significantly higher than the control group (0.184 ± 0.132) (t= 10.311,P< 0.001). Simultaneously, RT-qPCR and WB results indicated that the expression levels of AQP9 mRNA (0.327 ± 0.347) and protein levels (0.042 ± 0.004) in the experimental group + IGF-I group were significantly lower than the experimental group (t= 33.573, 5.598,P< 0.001, 0.005), suggesting that adipocyte co-culture had possibility to regulate the expression level of AQP9 through the PI3K-Akt pathway. Confocal laser microscopy analysis showed that the transfection efficiency was more than 90%. RT-q PCR and WB results indicated that the expression levels of AQP9 mRNA and protein levels (0.373 ± 0.221) in HepG2-AQP9 group were significantly higher than HepG2-PWPI group (t=14.953, 28.931,P= 0.002 and 0.000), suggesting that the stable overexpression of AQP9 cell line was successfully constructed. The intracellular lipid droplets and triglyceride content in HepG2-AQP9 co-culture group was significantly increased (t= 5.478, 5.369,P= 0.005) than HepG2-PWPI co-culture group and HepG2-AQP9 co-culture+ IGF-I group, suggesting that the increased expression of AQP9 had promoted HepG2 steatosis in co-cultured adipocytes. WB results showed the expression levels of p-Akt protein (0.168 ± 0.006) and p-Akt/total Akt (0.265±0.009) in HepG2-AQP9 co-culture + IGF-1 group was significantly increased (t= 16.311, 8.769,P< 0.001) than HepG2-AQP9 co-culture group, while the expression levels of AQP9 mRNA (0.327 ± 0.034) and protein (0.375 ± 0.025) was significantly decreased (t= 33.573, 9.146,P< 0.001 and 0.001). Conclusion: Adipocytes co-culture can induce steatosis in HepG2 cells, and may participate in inhibiting PI3K-Akt signaling pathway to upregulate the expression of AQP9 in steatotic HepG2 cells.

Anticardiolipin antibodies in various diseases in Taiwan: a retrospective analysis.

The goals of this study are to determine the frequency of anticardiolipin antibodies (ACA) in patients with various diseases and to evaluate the clinical significance of ACA in Taiwan. We collected 690 patients from ACA laboratory records. They were divided into eight groups in order to compare ACA percentages. Positive rates of ACA in different disease groups were below 20%, except for 38.2% in autoimmune diseases with vascular thrombosis. Compared with old stroke, the ACA positivity in young stroke was not significantly different (P = 0.482). The positive percentage of lupus anticoagulant (LA) (2.86%) was lower than that of ACA (15.66%) in young stroke (P = 0.015). Among patients with pregnancy loss or prematurity, the ACA positivity in lupus patients (44.44%) was higher than without lupus (9.76%; P = 0.01). The prevalence of ACA is higher in patients with vascular thrombosis complicated by autoimmune diseases than with thrombosis alone in Taiwan. Young and old stroke do not differ in ACA positivity. Moreover, ACA is more prevalent than LA for young stroke related coagulation. The ACA positivity for pregnancy loss or prematurity is very low in Taiwan. In summary, this is the first report on the frequency of ACA and other coagulation factors in various diseases in Taiwan.

Clinical experience of HIV/AIDS in a municipal hospital in Taiwan.

To describe the spectrum of epidemiological and major clinical manifestations of patients infected by human immunodeficiency virus type 1 (HIV-1) in a municipal hospital, a retrospective review was done of 53 HIV-1-infected patients who had been admitted to Taipei Municipal Jen-Ai Hospital between January 1990, and July 1996. The majority (94.3%) of the patients in the cohort were male. Peak incidence was found in the fourth decade (28.3%). Forty-four (83%) patients presented in the first hospital stay with acquired immunodeficiency syndrome (AIDS). The mean duration between establishment of diagnosis of HIV-1 infection and that of AIDS was 11.2 (0-84) months. Heterosexual transmission accounted for 54.7% of the infections in the study group, and bi-/homosexual men made up another 32%. Psychosis of new onset was noted in two patients. In all AIDS indicator conditions, Pneumocystis carinii pneumonia (PCP) was the leading opportunistic infection among AIDS patients. PCP was also on the top of initial manifestations of HIV-1 infection. One patient with Penicillium marneffei infection was diagnosed to have AIDS. The mean CD4 count at admission of AIDS patients was much lower than that of non-AIDS patients (32 vs. 297/microliter, p < 0.0005). During the follow-up period 24 of 53 patients died. Mean survival time of 23 expired patients after establishment of diagnosis of AIDS was 6.4 (0-29) months. The results indicated that males outnumbered females greatly in the number of cases. Sexual activity remained the most important route of infection. Psychosis of new onset may be an early manifestation of HIV-associated encephalopathy and requires more attention. In addition, the outcome was poor as most patients in this area did not become aware of risk of HIV-1 infection until they were seriously illed with full-blown AIDS that they would seek medical help. PCP was the most common incentive for medical consultation. Penicillium marneffei infection is endemic in southeast Asia, and should be classified as an AIDS indicator condition in Taiwan.

Clinical experience of esophageal ulcers and esophagitis in AIDS patients.

In Taiwan, numbers of patients with the acquired immunodeficiency syndrome (AIDS) have been increasing in recent years. We present esophageal disease of different causes in 5(16%) heterosexual men among 31 AIDS patients over a 5-year period. Major symptoms included mild dysphagia in 4 (80%) patients and odynophagia in 3 (60%) patients. The duration of symptoms varied from 3 days to 6 months. The symptoms occurred before the diagnosis of AIDS in 3 patients. At esophagogastroduodenoscopy (endoscopy), all 5 patients had esophagitis and/or esophageal ulcers proved by histopathologic evaluation. Four had Candida esophagitis, 3 had cytomegalovirus esophagitis/ulcers and 2 had idiopathic esophageal ulcerations (IEU). Three patients had different esophagitis/ulcers at the same time or during follow-up. The median CD4 lymphocyte count at the time of diagnosis of esophageal disease was 12.2 cells/mm3 (range, 3 to 35 cells/mm3). The endoscopic pictures of the different causes of esophagitis/ ulcers lack uniformity in number, size and appearance. These observations make a conclusion that all AIDS patients with an esophageal disease should undergo endoscopy with biopsy to obtain a definitive diagnosis.

Disseminated Penicillium marneffei infection in a patient with acquired immunodeficiency syndrome.

Penicillium marneffei is a rare human pathogen in Southeast Asia and Southern China. A 26-year-old heterosexual male had intermittent fever, diarrhea and weight loss for 3 months. He was severely immune deficient (CD4+ count, 4/microL) and serologic tests for antibodies against human immunodeficiency virus type-1 were positive. P. marneffei infection was disseminated with involvement of the liver, spleen and bone marrow. The diagnosis was established by histologic examination and confirmed by isolation of the fungus. This infection responded well to amphotericin B. He was discharged with maintenance therapy of itraconazole and continued to be stable during his 2-month follow-up.

Diagnostic value of plasma aldosterone/potassium ratio in hypoaldosteronism.

The diagnosis of hypoaldosteronism usually depends upon a combination of abnormal clinical and laboratory findings. The most common abnormality in hypoaldosteronism is hyperkalemia, which may be combined with sodium depletion. In the present study, 5 of 16 patients diagnosed with isolated hypoaldosteronism (IHA) had sodium depletion due to renal salt wasting, and four patients had normokalemia. Of these 16 IHA patients, 70% had subnormal baseline and stimulated plasma renin activity (PRA). Six patients diagnosed with type I pseudohypoaldosteronism (PHA) had normal or high PRA and plasma aldosterone concentrations (PAC). In 11 control subjects, supine PAC correlated positively with serum potassium (SK), and PAC stimulated by furosemide and ambulation correlated with the 24-hour urinary potassium excretion (UK). However, these correlations were not found in IHA and PHA patients. The ratio of UK/UNa+K and UNa/UK correlated with the stimulated PAC when the IHA and control subjects were taken as a whole. However, these electrolyte excretion parameters bore no relationship to the supine PAC. The stimulated PAC/SK ratio was used to discriminate the three groups; all IHA patients had a ratio below 3. The results indicate that stimulated PAC reflects the bioactivity of aldosterone on the collecting tubule, and the stimulated PAC/SK ratio is useful for the diagnosis of hypoaldosteronism and pseudohypoaldosteronism.

Acquired immunodeficiency syndrome with CNS toxoplasmosis: a case report.

Central nervous system (CNS) toxoplasmosis is an important infectious complication of acquired immunodeficiency syndrome (AIDS) which appears to result from reactivation of a previously acquired infection and requires prolonged treatment. A 31-year-old male presented in a drowsy mental state and with an unstable gait. Computerized tomographic (CT) scan and magnetic resonance imaging (MRI) showed multiple nodular lesions in the cerebrum and cerebellum; the seropositivity for the human immunodeficiency virus (HIV-1) and high serum IgG toxoplasma titers were also demonstrated. A presumptive diagnosis of CNS toxoplasmosis was based on neurological signs and neuroradiological findings. This was confirmed by improvement in both clinical and neuroradiological pictures during treatment with pyrimethamine and clindamycin. Four months later, however the patient died of intracranial hemorrhage and massive upper GI bleeding.

Serum thyrotropin-binding inhibiting immunoglobulin and thyroperoxidase antibody in Graves' hyperthyroidism after 131I therapy.

Seventeen patients who received radioiodine (131I) therapy for Graves' hyperthyroidism had serial blood samples taken before therapy and after therapy for a period of at least 1 year. At 1 year post-therapy, six patients were hypothyroid. Seven patients were euthyroid, and four patients were hyperthyroid. Prior to isotope administration, 14 patients had detectable serum thyrotropin-binding inhibiting immunoglobulin (TBII) and 16 patients had detectable serum thyroperoxidase antibody (TPOAb). Three to 6 months after therapy, 11 of 14 TBII-positive patients demonstrated a marked increase (> 10%) in serum TBII activity. Four patients out of 11 developed hypothyroidism and six of the 11 developed euthyroidism. A decrease in TBII was observed in three patients who developed hyperthyroidism. In the three patients with undetectable TBII prior to therapy, two had high titers of TPOAb. Seven patients demonstrated a marked increase in TPOAb 3 to 6 months after therapy. Of these, four developed hypothyroidism and three developed euthyroidism, whereas three of the four patients who had a marked decrease in TPOAb developed hyperthyroidism. This study demonstrated that an increase in serum TBII and TPOAb activity 3 to 6 months after 131I therapy, may be useful in predicting which patients may develop euthyroidism or hypothyroidism after 1 year of 131I therapy.

Acute muscular paralysis in an adult with subclinical Bartter's syndrome associated with gentamicin administration.

We report an adult case of asymptomatic Bartter's syndrome with the first presentation of hypokalemic paralysis triggered by gentamicin injection. Marked hypokalemia and hypomagnesemia associated with excessive kaliuresis and magnesiuria were found. Plasma renin activity and aldosterone concentration were high, but blood pressure was normal. Renal biopsy revealed hypercellularity of the renin-producing cell of the juxtaglomerular apparatus. Muscular paralysis subsided after potassium chloride supplementation. Hypokalemia was corrected with potassium and magnesium supplements and the use of diclofenac. To the best of our knowledge, there have been no reports of muscular paralysis associated with gentamicin in Bartter's syndrome.

Hormonal change in an adult with Prader-Willi syndrome: report of a case.

We report a classical case of Prader-Willi syndrome (PWS) in an adult with typical interstitial deletion of chromosome 15, and emphasize the study of hormonal change. This 21-year-old female had PWS face characteristics, small hands and feet, marked obesity, mental retardation, growth retardation, absence of puberty and amenorrhea. She also had the characteristic history of infantile hypotonia, poor feeding, failure to thrive and then improved appetite, followed by obesity from the age of four years. She had compulsive hyperphagia, to the extent of stealing and lying to take food. Chromosome study with high resolution banding technique revealed a small interstitial deletion at band q12 of chromosome 15, which is characteristically found in a majority of patients with PWS. Hormonal study revealed hypogonadism and growth hormone deficiency of supposed hypothalamic origin. She also had non-insulin-dependent diabetes mellitus with decreased pancreatic insulin reserve.

Prostatic ductal system in rats: regional variation in morphological and functional activities.

The rat prostate is a complex ductal system with branches and subbranches extending from one end to another. Owing to the relative distance of various regions of the duct from the urethra, the entire length of the ductal system can be arbitrarily divided into three segments, i.e., the proximal, intermediate, and distal segments. The present study was carried out to assess the regional variation in cellular activities in this ductal system. Ventral prostates from adult Sprague-Dawley rats were dissected so that an individual ductal system was mechanically isolated and longitudinally sectioned to reveal various segments. Epithelial cells lining distal segments were tall-columnar type and were actively engaging in mitotic activity. Cells in intermediate segments were also tall-columnar type. However, they were mitotically quiescent, but able to produce secretory proteins. Evidence of programmed cell death was not observed in either of these two segments. Cells in proximal segments, on the other hand, were low-columnar or cuboidal in shape and were stained heavily for cathepsin D, a marker associated with late manifestation of cell death. Following castration in adult rats, there was a reversal in the site of programmed death in cells lining the ductal system. By Day 4 post-castration, distal segments contained many epithelial cells with intense cytoplasmic staining for cathepsin D while proximal segments showed a reduction in number of positively stained cells. By Day 7 post-castration, cells in proximal segments, though atrophied, were devoid of staining for cathepsin D.(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of gene expression in rat prostate by androgen and beta-adrenergic receptor pathways.

Denervation of rat ventral prostate has been accomplished by excising prostatic tissue fragments and implanting them under the renal capsules of intact syngeneic rats. This resulted in a substantial reduction of expression of a major organ-specific secretory protein, prostatic binding protein (PBP). The depressed level of PBP and its subunits and mRNAs could be restored, however, to as much as 80% of control levels by the administration of a pharmacological dose of exogenous androgen, testosterone propionate (TP), and/or a beta-adrenergic agonist, isoproterenol (ISO). Furthermore, compared to ascorbate-treated controls, TP and ISO increased the synthesis of total cellular protein and PBP by the prostatic renal implants. TP and/or ISO also remodelled the luminal epithelial structure and elevated secretory functions. ISO alone had no effect, however, in castrated animals, indicating that androgen plays a dominant role in the restoration of tissue PBP content. Concomitant to increased PBP content and remodelling of prostatic histomorphology, androgen was also found to raise the depressed levels of beta 2-adrenergic and androgen receptors in the prostatic isografts maintained in intact hosts. In contrast, although an established rat prostatic epithelial cell line (NbE-1) contains high affinity androgen receptor, androgen failed to restore beta-adrenergic receptor as well as PBP content in this cultured cell line. These results, taken together, suggest that a tight coupling between androgen receptor and beta 2-adrenergic receptor pathways may be a prerequisite for PBP expression and functional differentiation in the rat ventral prostate gland.

Inter-tissue and inter-species characteristics of the mitochondrial carnitine palmitoyltransferase enzyme system.

Properties of the carnitine palmitoyltransferase (EC 2.3.1.21) (CPT) enzyme system were compared in isolated mitochondria from a range of tissues in rodents, monkey, and man. Common features were as follows: (a) while membrane-bound, CPT I, but not CPT II, was inhibited reversibly by malonyl-coenzyme A (CoA) and irreversibly by CoA esters of certain oxirane carboxylic acids; (b) the detergent, Tween-20, readily solubilized CPT II in active form while leaving CPT I membrane associated and catalytically functional; (c) octyl glucoside and Triton X-100 released active CPT II but caused essentially complete loss of CPT I activity. Use of [3H]tetradecylglycidyl-CoA, a covalent ligand for CPT I, yielded estimates of the enzyme's monomeric molecular size: approximately 86 kDa in non-hepatic tissues and approximately 90-94 kDa in liver, depending upon species. A polyclonal antibody to purified rat liver CPT II recognized a single protein in each tissue; its apparent molecular mass was approximately 70 kDa in all rat tissues and approximately 68 kDa in all mouse tissues as well as monkey and human liver. On Northern blot analysis a rat liver CPT II cDNA probe detected a single approximately 2.5-kilobase mRNA in all rat and mouse tissues examined. The following points are emphasized. First, CPT I and II are different proteins. Second, within a species CPT II, but not CPT I, is probably conserved across tissue lines. Third, slight variations in size of both enzymes were found in different species, although, at least in the case of CPT II, significant amino acid identity exists among the various isoforms. Fourth, CPT I, unlike CPT II, requires membrane integrity for catalytic function. Finally, the strategic use of detergents provides a simple means of discriminating between the two enzyme activities.

Apolipoproteins A-I and B in non-insulin-dependent diabetes mellitus.

In recent years apolipoproteins A-I and B examinations have been performed on patients with coronary artery disease as a better predictor of the severity of atherosclerosis. In the present study, 21 treated male and 22 treated female patients with non-insulin-dependent diabetes mellitus (NIDDM) were examined and compared with controls of the same sex, age and body mass (23 males, 21 females). Cholesterol, triglyceride, LDL-cholesterol in male and female patients with NIDDM were significantly higher than in male and female controls. HDL-cholesterol in male and female patients with NIDDM was not different from those of male and female controls. Apolipoproteins A-I and B in male and female patients with NIDDM were higher than in male and female controls. [Apolipoproteins A-I (g/L) male 1.40 +/- 0.21 vs 1.25 +/- 0.15, p less than 0.005; female 1.56 +/- 0.23 vs 1.42 +/- 0.24, p less than 0.025. Apolipoproteins B (g/L) male 1.29 +/- 0.30 vs 0.97 +/- 0.22, p less than 0.001; female 1.34 +/- 0.34 vs 0.98 +/- 0.35, p less than 0.001.] Discrepancy between the higher apolipoprotein A-I and the normal HDL-cholesterol in in NIDDM supports the theory of altered composition of HDL particles in diabetic patients. The controversy between the higher apolipoprotein A-I and the higher incidence of atherosclerosis in patients with NIDDM makes the clinical usefulness of this laboratory measurement doubtful in these patients.

Fusion proteins containing androgen receptor sequences and their use in the production of poly- and monoclonal anti-androgen receptor antibodies.

Complementary DNA segments that encode different domains of human and rat androgen receptors were fused to the Escherichia coli trpE gene using pATH expression vectors. Fusion proteins expressed by the bacteria were used to immunize rats and rabbits to obtain polyclonal antibodies to androgen receptors. Spleen cells of immunized rats were fused with myeloma cells to obtain stable hybridomas that produced monoclonal antibodies. Gradient centrifugation and immuno-precipitation assays indicated that the antibodies interacted with androgen receptors specifically.

Infectivity in asymptomatic hepatitis B carriers: its correlation with HBeAg/anti-HBe status.

Sera collected from 190 asymptomatic HBsAg carriers with varying status of HBe markers were tested for HBV DNA using the slot-blot hybridization method and the results were compared with serum alanine aminotransferase (ALT) levels. No significant difference was observed in the positive rate of serum HBV DNA between patients showing either HBeAg (+)/normal ALT or HBeAg (+)/high ALT. On the other hand, in cases with positive anti-HBe or negative for both HBeAg and anti-HBe, statistically significant differences could be shown in HBV DNA positivity between normal and high ALT subjects. In the group of 100 patients possessing anti-HBe antibody, 50% of the cases with high ALT levels were positive for serum HBV DNA, whereas all of the cases with normal ALT levels were negative for serum HBV DNA (p less than 0.0002). In 21 patients showing HBeAg (-)/anti-HBe(-), HBV DNA could be detected in the serum of 60% of the cases with high ALT levels, but in none of the cases with normal ALT levels (p less than 0.05). Our results suggest that HBeAg alone is a reliable marker in the prediction of infectivity in asymptomatic carriers. In cases showing anti-HBe (+), loss of infectivity could be ascertained if the patient had normal serum ALT, but, continuous viral replication should be suspected in cases with high serum ALT values.

Identification of an androgen-repressed mRNA in rat ventral prostate as coding for sulphated glycoprotein 2 by cDNA cloning and sequence analysis.

The concentrations of a small number of mRNAs in the rat ventral prostate increase after castration and then decrease upon androgen treatment. Since the repression of specific gene expression may be important in the regulation of organ growth, we have cloned a cDNA for an androgen-repressed mRNA, the concentration of which increased 17-fold 4 days after castration, and this increase was reversed rapidly by androgen treatment. By sequence analysis the androgen-repressed mRNA was identified as that coding for sulphated glycoprotein 2.

Intracellular inhibition of chromatin binding and transformation of androgen receptor by 3'-deoxyadenosine.

Incubation of minced rat ventral prostate with 3'-deoxyadenosine (3'-dA) prior to labeling with the androgen, tritiated 7 alpha, 17 alpha-dimethyl-19-nortestosterone, reduced the level of androgen receptor bound to chromatin and increased the level of cytosolic androgen receptor and the fraction of cytosolic androgen receptor that did not bind to DNA. This effect was specific for 3'-dA and not mimicked by adenosine, 2'-deoxy-adenosine, cytidine, guanosine, or uridine. Adenosine was a competitive inhibitor of the 3'-dA effect. Labeled cytosolic androgen receptor from 3'-dA-treated prostate had properties that were similar to those exhibited by untransformed androgen receptor from prostate cytosol prepared in the presence of Na2MoO4, an inhibitor of receptor transformation in cell-free systems. Both androgen receptors had sedimentation coefficients of 8-9 S in low-salt gradients, did not bind to DNA tightly, and had a high affinity for DEAE-cellulose. The 3'-dA effect on these properties was not observed if androgen receptor from 3'-dA-treated prostate was isolated on high-salt gradients. These findings show that androgen receptor transformation does take place in intact prostate cells and suggest that 3'-dA inhibits chromatin binding of androgen receptor by interfering with androgen receptor transformation. The transformation process appears to involve removal of components from androgen receptor. Since 3'-dA is a potent inhibitor of the synthesis, polyadenylation, and nucleocytoplasmic transport of RNA, the 3'-dA effect may indicate a role for RNA in the mechanism of receptor transformation in intact target cells.

Structural analysis of complementary DNA and amino acid sequences of human and rat androgen receptors.

Structural analysis of cDNAs for human and rat androgen receptors (ARs) indicates that the amino-terminal regions of ARs are rich in oligo- and poly(amino acid) motifs as in some homeotic genes. The human AR has a long stretch of repeated glycines, whereas rat AR has a long stretch of glutamines. There is a considerable sequence similarity among ARs and the receptors for glucocorticoids, progestins, and mineralocorticoids within the steroid-binding domains. The cysteine-rich DNA-binding domains are well conserved. Translation of mRNA transcribed from AR cDNAs yielded 94- and 76-kDa proteins and smaller forms that bind to DNA and have high affinity toward androgens. These rat or human ARs were recognized by human autoantibodies to natural ARs. Molecular hybridization studies, using AR cDNAs as probes, indicated that the ventral prostate and other male accessory organs are rich in AR mRNA and that the production of AR mRNA in the target organs may be autoregulated by androgens.