Correlation of tumor suppressor P53 RNA expression with human immunodeficiency virus disease in rapid and slow progressors

OBJECTIVE: To determine the relation between P53 tumor suppressor RNA expression and human immunodeficiency virus (HIV) disease progression. STUDY DESIGN/METHODS: A quantitative assay of P53 RNA expression was used to analyze a cohort of HIV-negative persons. The assay was then used in longitudinal and cross-sectional studies of HIV slow and rapid progressors. RESULTS: We demonstrate first that P53 expression in peripheral blood mononuclear cells from HIV-1-seronegative persons is minimal. Longitudinal studies in a small cohort of HIV-1-infected slow and rapid progressors reveal that rapid progressors seem to have greater P53 RNA expression over time. This was validated in a cohort of 26 HIV-1-infected persons in whom the expression of P53 RNA was significantly greater in persons with rapid progression of HIV-1 disease. CONCLUSION: These data suggest that P53 RNA expression may play a role in the pathogenesis of HIV-1 disease, though the mechanism of this interaction remains unknown.     

Behavioral characteristics of a mouse model of cancer pain

Pain is a major symptom in cancer patients, and most cancer patients with advanced or terminal cancers suffer from chronic pain related to treatment failure and/or tumor progression. In the present study, we examined the development of cancer pain in mice. Murine hepatocarcinoma cells, HCa-1, were inoculated unilaterally into the thigh or the dorsum of the foot of male C3H/HeJ mice. Four weeks after inoculation, behavioral signs were observed for mechanical allodynia, cold allodynia, and hyperalgesia using a von Frey filament, acetone, and radiant heat, respectively. Bone invasion by the tumor commenced from 7 days after inoculation of tumor cells and was evident from 14 days after inoculation. Cold allodynia but neither mechanical allodynia nor hyperalgesia was observed in mice that received an inoculation into the thigh. On the contrary, mechanical allodynia and cold allodynia, but not hyperalgesia, were developed in mice with an inoculation into the foot. Sometimes, mirror-image pain was developed in these animals. These results suggest that carcinoma cells injected into the foot of mice may develop severe chronic pain related to cancer. This animal model of pain would be useful to elucidate the mechanisms of cancer pain in humans.     

Overexpression of decoy receptor 3 in precancerous lesions and adenocarcinoma of the esophagus

Overexpression of decoy receptor (DcR) 3 protein, a recently discovered member of the tumor necrosis factor receptor superfamily, was examined in 40 esophagogastrectomy specimens containing areas of Barrett esophagus (n = 27), low-grade dysplasia (n = 27), high-grade dysplasia or carcinoma in situ (n = 22), and esophageal adenocarcinoma (EAC; n = 28) with immunohistochemical analysis. The results revealed significantly more overexpression of DcR3 in high-grade dysplasia or carcinoma in situ and EAC than in benign esophageal mucosa (both P < .0001), Barrett esophagus (both P < .001), and low-grade dysplasia (P < .01 and P = .033, respectively). Low-grade dysplasia also showed significant overexpression of DcR3 compared with benign esophagus (P < .05) but not with Barrett esophagus (P > .05). DcR3 overexpression seems to negatively correlate with the grade of EAC. Our results suggest that overexpression of DcR3 protein might aid in the diagnosis of high-grade dysplasia or carcinoma in situ and EAC and also might serve as a potential therapeutic target.     

Biomechanical assessment with electromyography of post-stroke ankle plantar flexor spasticity

Spasticity has been defined as a motor disorder characterized by a velocity-dependent increase in tonic stretch reflex (muscle tone). Muscle tone consists of mechanical-elastic characteristics, reflex muscle contraction and other elements. The aims of this study were to determine whether to assess spasticity quantitatively, and to characterize biomechanical and electromyographic spasticity assessment parameters. These assessment parameters were described by investigating the correlation between clinical measures and the response to passive sinusoidal movement with consecutive velocity increments. Twenty post-stroke hemiplegic patients and twenty normal healthy volunteers were included in the study. Five consecutive sinusoidal passive movements of the ankle were performed at specific velocities (60, 120, 180, and 240 degrees/ sec). We recorded the peak torque, work, and threshold angle using a computerized isokinetic dynamometer, and simultaneously measured the rectified integrated electromyographic activity. We compared these parameters both between groups and between different velocities. The peak torque, threshold angle, work, and rectified integrated electromyographic activity were significantly higher in the post-stroke spastic group at all angular velocities than in the normal control group. The threshold angle and integrated electromyographic activity increased significantly and linearly as angular velocity increased, but the peak torque and work were not increased in the post-stroke spastic group. Peak torque, work, and threshold angle were significantly correlated to the Modified Ashworth scale, but the integrated electromyographic activity was not. The biomechanical and electromyographic approach may be useful to quantitatively assess spasticity. However, it may also be very important to consider the different characteristics of each biomechanical parameter.     

A case of leukemia-associated arthritis--identification of leukemic cells in synovial fluid by light microscopy

One case of arthritis complicating leukemia is described in which leukemic cells were identified in synovial fluid by light microscopy. Although arthritis is a well-known manifestation of leukemia with an incidence of 13.5%, the pathogenesis often is unclear, and the direct demonstration of leukemic cells in synovial fluid has been very uncommon. A 16 year-old male patient was admitted due to left elbow joint pain and swelling. Synovial fluid examination revealed blast cells and this finding has directed to a final diagnosis of acute lymphoblastic leukemia.     

Premenstrual anxiety and depression: comparison of objective psychological tests with a retrospective questionnaire

Increased anxiety and depression are among the most frequently reported psychological problems in women seeking help for severe symptomatic premenstrual change, but there has been little objective evaluation of these symptoms. We therefore examined the results of objective psychological testing in 40 women with no apparent psychiatric or psychological disorder who had reported moderate to extreme increased anxiety and depression on a retrospective assessment form. Scores on the State-Trait Anxiety Inventory and Institute of Personality and Ability Testing (IPAT) Depression Scale increased from the low symptom intermenstrual phase of the cycle (days 5-10) to the premenstrual phase (within the last 6 days of the cycle), suggesting that retrospective complaints of increased premenstrual anxiety and depression can be confirmed on objective psychological assessment. However, it was observed that the distribution of intermenstrual IPAT depression scores was bimodal. Cyclic changes varied among the tests depending upon the IPAT depression score. The study suggests that 2 populations may exist in this screened sample; one population appears to have "pure PMS" and the second groups manifests a premenstrual exacerbation of subclinical depression.     

Induction of rat liver microsomal epoxide hydrolase by thiazole and pyrazine: hydrolysis of 2-cyanoethylene oxide

Liver microsomal epoxide hydrolase (mEH) is active in the detoxificationof epoxide-containing carcinogens. The effects of thiazole andpyrazine, constituents of tobacco and tobacco smoke as wellas of a variety of foods, on the expression and regulation ofmEH were examined in rats (200 mg/kg body wt/day, i.p., 1/emdash3 days). Immunoblot analyses using rabbit anti-rat mEH antibodyrevealed a significant increase in mEH levels in hepatic microsomesisolated from either thiazole- or pyrazine-treated animals.Another protein (43 kd) cross-reacting with polyclonal mEH antibodywas found to be increased concomitantly following pyrazine treatment.Northern and slot blot analyses showed substantial increasesin mEH mRNA following either thiazole or pyrazine treatment.The level of mEH mRNA increased 17-fold at 24 h following thiazoletreatment, relative to control. Approximately 20- and 16-foldincreases in mEH mRNA were also observed at 48 and 72 h respectivelyfollowing treatment with pyrazine. The level of polymerase chainreaction (PCR)-amplified mEH DNA derived from poly(A)+ RNA wasclearly elevated following either thiazole or pyrazine treatmentrelative to that from untreated animals. Both sense and antisensestrands of PCR-amplified mEH DNA were cloned into an M13mpl9phage vector in order to examine the nucleotide sequences ofPCR-amplified mEH DNA derived from the poly(A)+ RNA isolatedfrom thiazole- or pyrazine-treated animals. Sequence analysesrevealed that the sequence of PCR-amplified DNA from the inducedmRNA was identical to that published for mEH cDNA. Epoxide hydrolaseactivity toward the hydrolysis of 2-cyanoethylene oxide (CEO),the epoxide metabolite of the rat carcinogen acrylonitrile,was not significant in hepatic microsomes from untreated rats,but was substantially induced by treatment with thiazole orpyrazine. Microsomal hydrolysis activity was heat-sensitiveand potently inhibited by l, l, l-trichloropropene-2, 3-oxide,indicating that mEH was the catalyst. The V_max for the hydrolysisof CEO by hepatic microsomes from thiazole-treated rats (13.4nmol/min/mg protein) was 1.5-fold greater than that with microsomesfrom pyrazine-treated rats, whereas similar K_m values ( 1 mM)were observed for both microsomal preparations. These kineticdata correlate well with the increases in mEH mRNA observedafter administration of thiazole or pyrazine to rats. Theseresults provide evidence that administration of thiazole orpyrazine induces mEH with a large increase in mEH mRNA, andthat the induced mEH catalyzes the hydrolysis of CEO.     

Structure activity relationship of ar-turmerone analogues

For the analysis of structure activity relationship of ar-turmerone analogues, the compounds containing the various substituents on the phenyl ring and 1(or 2)-naphthyl group in the place of phenyl of ar-turmerone were prepared and tested their cytotoxicity against HL-60, K-562, and L1210 leukemia cellsin vitro. The substituents at para position are methoxy, phenoxy, methyl, trifluoromethyl, fluoro, and chloro. Atmeta position methoxy, methyl, trifluoromethyl, or chloro groups and atortho position methoxy or chloro group were introduced. Against HL-60 and K-562 cells, ED₅₀ values of the analogues are ranged from 0.8 to 30.0 μg/ml. Against L1210 cell, these are located more than 20.0 μg/ml. However, 5-carboethoxy-2-methyl-6-(1-naphthyl)-2-octen-4-one (5n) possesses ED50 valuses 0.8, 2.1, 6.5 μg/ml against HL-60, K-562, L1210 cells, respectively. The electronic nature of the subsituents on phenyl ring of ar-turmerone dose not affect the biological activity. Therefore the flat structure of aromatic portion of ar-turmerone analogues is the more important factor for their activity rather than its electronic nature. The potentiation of the cytotoxicity with the enlargement of aromatic ring region also supports the importance of the plane structure of this area. The restriction of the single bond rotation between C-6 and aromatic ring through the introduction of substituents at theortho position of phenyl ring and the increment of size of alkyl group at C-6 position enhances the activity. Therefore the effective conformation should be the one having the orthogonal arrangement between the aromatic ring and the side chain.     

Characterization of the bisphosphonate recognition site on hydroxyapatite using radioligand binding techniques with [14C]citric acid

Summary The present studies characterize the binding of [¹⁴C]citric acid to synthetic hydroxyapatite (HA) crystals. [¹⁴C]Citric acid specifically bound to HA and was dependent upon the concentration of HA in the assay. The binding of [¹⁴C]citric acid to HA reached equilibrium within 20 min and remained stable for at least 90 min. Dissociation of bound [¹⁴C]citric acid was biphasic in nature since both rapid and more slowly reversible binding components were detected. Saturation experiments also indicated that [¹⁴C]citric acid labeled two recognition sites with different affinity (Kd_H=42 nM and Kd_L=24,000 nM) and density (Bmax_H=161 fmol/g HA and Bmax_L=8.8 pmol/g HA). Ligand competition experiments revealed that compounds that are known to readily bind bone (e.g., sodium pyrophosphate, methylene diphosphonic acid, etidronate) potently inhibited the binding of [¹⁴C]citric acid to HA, whereas compounds known to have poorer affinity for bone (e.g., oxalic acid and GABA) did not. Computer analysis of these inhibition curves revealed specific ligand interactions at two different affinity recognition sites. The present results indicate that [¹⁴C]citric acid binds discrete sites on synthetic HA in a fashion consistent with a specific labeling of the bisphosphonate recognition site. Analysis of the binding of [¹⁴C]citric acid to HA provides a useful method to further explore the structure activity relationships of novel compounds that have binding affinity for bone.