Mediation of oxidative stress in hypothalamic ghrelin-associated appetite control in rats treated with phenylpropanolamine.

Phenylpropanolamine (PPA)-induced appetite control is associated with oxidative stress in the hypothalamus. This study explored whether hypothalamic antioxidants participated in hypothalamic ghrelin system-associated appetite control in PPA-treated rats. Rats were given PPA daily for 4 days, and changes in food intake and the expression of neuropeptide Y (NPY), the cocaine- and amphetamine-regulated transcript (CART), superoxide dismutase, catalase, ghrelin, acyl ghrelin (AG), ghrelin O-acyltransferase (GOAT) and the ghrelin receptor (GHSR1a) were examined and compared. Results showed that both food intake and the expression of NPY and ghrelin/AG/GOAT/GHSR1a decreased in response to PPA treatment with maximum decrease on Day 2 of the treatment. In contrast, the expression of antioxidants and CART increased, with the maximum increase on Day 2, with the expression opposite to that of NPY and ghrelin. A cerebral infusion of either a GHSR1a antagonist or reactive oxygen species scavenger modulated feeding behavior and NPY, CART, antioxidants and ghrelin system expression, showing the involvement of ghrelin signaling and oxidative stress in regulating PPA-mediated appetite control. We suggest that hypothalamic ghrelin signaling system, with the help of antioxidants, may participate in NPY/CART-mediated appetite control in PPA-treated rats.

All-optically controllable random laser based on a dye-doped polymer-dispersed liquid crystal with nano-sized droplets.

This study elucidates for the first time an all-optically controllable random laser in a dye-doped polymer-dispersed liquid crystal (DDPDLC) with nano-sized LC droplets. Experimental results demonstrate that the lasing intensity of the random laser can be controlled to decrease by increasing irradiation time/intensity of one green beam, and increase by increasing the irradiation time of one red beam. The all-optical controllability of the random laser is attributed to the green (red)-beaminduced isothermal nematic-->isotropic (isotropic-->nematic) phase transition in LC droplets by trans-->cis (cis-->trans back) isomerization of azo dyes. This isomerization may decrease (increase) the difference between the refractive indices of the LC droplets and the polymer, thereby increasing (decreasing) the diffusion constant (or transport mean free path), subsequently decreasing the scattering strength and, thus, random lasing intensity.

Spatial and polarization entanglement of lasing patterns and related dynamic behaviors in laser-diode-pumped solid-state lasers.

To provide the underlying physical mechanism for formations of spatial- and polarization-entangled lasing patterns (namely, SPEPs), we performed experiments using a c-cut Nd:GdVO(4) microchip laser with off-axis laser-diode pumping. This extends recent work on entangled lasing pattern generation from an isotropic laser, where such a pattern was explained only in terms of generalized coherent states (GCSs) formed by mathematical manipulation. Here, we show that polarization-resolved transverse patterns can be well explained by the transverse mode-locking of distinct orthogonal linearly polarized Ince-Gauss (IG) mode pairs rather than GCSs. Dynamic properties of SPEPs were experimentally examined in both free-running and modulated conditions to identify long-term correlations of IG mode pairs over time. The complete chaos synchronization among IG mode pairs subjected to external perturbation is also demonstrated.

Luteolin induces apoptosis in oral squamous cancer cells.

Oral squamous cell carcinoma is the most common malignancy of the oral cavity, and treatment approaches are inadequate. Luteolin, a natural flavonoid compound, has been shown to have anti-tumorigenic properties on various types of tumors. Therefore, we hypothesized that luteolin has anti-tumorigenic properties for oral squamous cell carcinoma, and may provide effective chemotherapy. Results revealed that luteolin reduced the viability of SCC-4 cells and induced apoptosis by decreasing the expression of cyclin-dependent kinase (CDKs), cyclins, and phosphor- retinoblastoma (p-Rb) anti-apoptotic protein, but increased the expression of pro-apoptotic proteins and activated caspase 9 and 3, with a concomitant increase in the levels of cleaved poly-ADP-ribose polymerase (PARP). Combination treatment of luteolin with paclitaxel enhanced the cytotoxic effect of paclitaxel in SCC-4 cells, and continuous administration of luteolin suppressed the growth of xenograft tumors in nude mice. These results suggest that luteolin could be an effective chemotherapeutic agent for the treatment of oral squamous cell carcinoma.

Translocation between chromosome 5q35 and chromosome 11q13-- an unusual cytogenetic finding in a primary refractory acute myeloid leukemia.

Cytogenetic abnormalities are observed in approximately two-thirds of patients with acute myeloid leukemia (AML). Chromosome rearrangements are associated with specific subtypes of AML and associated prognosis. We report a patient with AML, M2, who was primarily refractory to standard induction chemotherapy with idarubicin and cytarabine. Flow cytometry of a bone marrow aspirate showed aberrant expression of B-cell markers including CD19. Cytogenetic studies disclosed a translocation between 5q35 and 11q13. Fluorescence in situ hybridization analyses demonstrated that neither the NSD1 nor MLL genes were involved in this case. Further study is required to define conclusively the genes involved and their contribution to pathogenesis in this case.

Silibinin inhibits invasion of oral cancer cells by suppressing the MAPK pathway.

Oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral cavity. Here, we provide molecular evidence associated with the anti-metastatic effect of silibinin by showing a marked inhibition of the invasion and motility of SCC-4 tongue cancer cells, with 89% and 66.4% of inhibition, respectively, by 100 microM of silibinin. This effect was associated with a reduced expression of MMP-2 and u-PA, together with an enhanced expression of TIMP-2 and PAI-1. Silibinin also exerted an inhibitory effect on the phosphorylation of ERK1/2. Additionally, pre-treatment of SCC-4 cancer cells with 10 and 20 microM of U0126, a specific MEK inhibitor, resulted in a reduced expression of MMP-2 (18.7 and 51.4%) and u-PA (19.2 and 48.9%) concomitantly with a marked inhibition of cell invasion (13.7 and 45.7%). Finally, silibinin was evidenced by its inhibition of the metastasis of Lewis lung carcinoma (LLC) cells in vivo. These results suggested that silibinin can reduce the invasion and metastasis of tumor cells, and such a characteristic may be of great value in the development of a potential cancer therapy.

Regulation of interleukin-6 expression by arecoline in human buccal mucosal fibroblasts is related to intracellular glutathione levels.

OBJECTIVES: Cytokines play an important role in regulating fibroblast function and is likely to play a key role in regulating the initiation and progression of scarring in any fibrotic disease. Interleukin-6 (IL-6) has been implicated in the development of a variety of fibrotic diseases. The aim of this study was to compare IL-6 expression in fibroblasts cultured from normal human buccal mucosa and oral submucous fibrosis (OSF) specimens and further explore the potential mechanism that may lead to induce IL-6 expression. METHODS: mRNA level of IL-6 in fibroblasts from OSF was compared with normal buccal mucosa. The effects of arecoline, the major areca nut alkaloid, on IL-6 expression in normal human buccal mucosa fibroblasts (BMFs) were measured in vitro. mRNA was quantified with AlphaImager 2000. To determine whether glutathione (GSH) levels were important in the induction of IL-6 by arecoline, we pretreated cells with 2-oxothiazolidine-4-carboxylic acid (OTZ) to boost GSH levels or with buthionine sulfoximine (BSO) to deplete GSH. RESULTS: Fibroblasts derived from OSF exhibited higher IL-6 gene expression than BMF in mRNA levels (P < 0.05). The exposure of quiescent BMF to arecoline resulted in the elevation of IL-6 mRNA expression in a dose-dependent manner (P < 0.05). IL-6 gene regulated by arecoline correlated with intracellular GSH levels in BMF. Arecoline at a concentration of 129 muM induced about 2.7-fold IL-6 mRNA levels over the 6-h incubation period. However, BSO enhanced the IL-6 mRNA levels by 3.9-fold (P < 0.05). In addition, OTZ was found to marginally reduce the arecoline-induced IL-6 expression by about 1.7-fold (P < 0.05). CONCLUSIONS: Taken together, these results suggest that IL-6 expression is significantly upregulated in OSF fibroblasts in areca quid chewers and arecoline may be responsible for the enhanced IL-6 expression. In addition, the regulation of IL-6 expression induced by arecoline is critically dependent on the intracellular GSH concentrations.

Alternations in quantities and activities of erythrocyte cytosolic carbonic anhydrase isoenzymes in glucose-6-phosphate dehydrogenase-deficient individuals.

BACKGROUND: This study was designed to evaluate the quantitative and activity alterations of cytosolic carbonic anhydrase (CA) isoenzymes in the erythrocytes of glucose-6-phosphate dehydrogenase (G6PD)-deficient individuals. METHODS: Western Blot and CA esterase activity analysis were employed to measure cytosolic erythrocyte CA isoenzymes. RESULTS: The total CA activities were analyzed from erythrocytes of 30 healthy and 30 G6PD-deficient individuals. The mean values with standard error (SE) were 22.9+/-1.69 U/gHb and 27.2+/-2.1 U/gHb (P<0.01), respectively. The ratio of CAI/CAII of G6PD-deficient individuals (1.28+/-0.06) was significantly lower than that of the normal subjects (3.79+/-0.18) (P<0.001). Furthermore, the concentration of CAIII in G6PD-deficient individuals was significantly lower than that of the normal subjects (P<0.001) and there were significant correlations between the concentration of CAI, CAII, CAIII, and ratio of CAI/CAII, and the activity concentration of G6PD. CONCLUSIONS: Different carbonic anhydrase isoenzymes may serve different roles in the G6PD-deficient erythrocyte. CAI could be used as an indicator for hemolytic anemia. CAII is able to compensate for the functions of CAI and increased expression of CAII will promote oxidative damage. CAIII can provide the G6PD-deficient persons with some extent of protection against oxidative damage.

Three-dimensional laser microvision.

A three-dimensional (3-D) optical imaging system offering high resolution in all three dimensions, requiring minimum manipulation and capable of real-time operation, is presented. The system derives its capabilities from use of the superstructure grating laser source in the implementation of a laser step frequency radar for depth information acquisition. A synthetic aperture radar technique was also used to further enhance its lateral resolution as well as extend the depth of focus. High-speed operation was made possible by a dual computer system consisting of a host and a remote microcomputer supported by a dual-channel Small Computer System Interface parallel data transfer system. The system is capable of operating near real time. The 3-D display of a tunneling diode, a microwave integrated circuit, and a see-through image taken by the system operating near real time are included. The depth resolution is 40 mum; lateral resolution with a synthetic aperture approach is a fraction of a micrometer and that without it is approximately 10 mum.

Lymphangioleiomyomatosis: abdominopelvic CT and US findings.

PURPOSE: To describe the abdominal computed tomographic (CT) and ultrasonographic (US) findings in patients with thoracic lymphangioleiomyomatosis (LAM) and to relate the prevalence of the findings to the severity of pulmonary disease. MATERIALS AND METHODS: Eighty patients with LAM underwent chest and abdominopelvic CT and abdominopelvic US. The images were reviewed prospectively by one radiologist, and the abdominal findings were recorded and correlated with the severity of pulmonary disease at thin-section CT. RESULTS: Sixty-one (76%) of 80 patients had positive abdominal findings. The most common abdominal findings included renal angiomyolipoma (AML) in 43 patients (54%), enlarged abdominal lymph nodes in 31 (39%), and lymphangiomyoma in 13 (16%). Less common findings included ascites in eight (10%), dilatation of the thoracic duct in seven (9%), and hepatic AML in three (4%). A significant correlation (P =.02) was observed between enlarged abdominal lymph nodes and increased severity of lung disease. CONCLUSION: There are characteristic abdominal findings in patients with LAM that, in conjunction with the classic thin-section CT finding of pulmonary cysts, are useful in establishing this diagnosis.

Significant differences in serum activities of matrix metalloproteinase-2 and -9 between HCV- and HBV-infected patients and carriers.

To examine the possible involvement of MMP-9 and -2 in the development of liver diseases caused by HCV or HBV infection, serum activities of both enzymes were studied by zymograph. Eight groups of subjects (60 for each) were examined in the study: healthy control, patients with hepatoma, liver cirrhosis, chronic hepatitis B or chronic hepatitis C, and carriers positive for HBsAg, both HBsAg and HBeAg, or anti-HCV. The results showed significant changes in the MMP-9 and -2 activities in the carriers. The presence of HBeAg was accompanied by a highest activity of MMP-2 and an inversely correlated (r=-0.578, P=<0.001), lowest activity of MMP-9 among all groups. For those with active liver diseases, MMPs activities were fluctuated at each stage of pathological symptoms. Chronic hepatitis B and C patients had significant different serum MMP-2 and -9 activities. These findings imply an influence on the balance of MMPs system by the existence of virus that might influence the following progression of liver disease, and a distinction between the pathological mechanisms of HCV and HBV. Since the serum MMPs activities were significantly varied between each stage of liver disease, an individual profile of these parameters might serve as an easy accessing serum marker to monitor the progression of liver disease.

Pulmonary lymphangioleiomyomatosis: correlation of ventilation-perfusion scintigraphy, chest radiography, and CT with pulmonary function tests.

PURPOSE: To determine the findings on ventilation-perfusion (V-P) scintigrams, computed tomographic (CT) scans, and chest radiographs and correlate them with pulmonary function test results in patients with lymphangioleiomyomatosis. MATERIALS AND METHODS: V-P scintigraphy, chest radiography, conventional and thin-section CT, and pulmonary function tests were performed in 39 patients. The images were graded on a scale of 0 (normal) to 3 (severely abnormal). RESULTS: Imaging abnormalities were found on 92% of ventilation scintigrams, 92% of perfusion scintigrams, 79% of chest radiographs, 100% of CT scans, and 100% of thin-section CT scans. On ventilation scintigrams, 28 (72%) patients demonstrated a speckling pattern. On CT scans, all patients had pulmonary cysts. Univariate analysis showed that extent of disease on chest radiographs and CT scans, cyst size, V-P abnormalities, and degree of speckling were inversely correlated with forced expiratory volume in one second (FEV(1)), diffusing capacity of lung for carbon monoxide, and the ratio of FEV(1) to forced vital capacity (FVC) (P <.01) but not with FVC and total lung capacity. Larger cyst size correlated with extent of disease at CT, but not significantly (P =.056). CONCLUSION: Scintigraphic and radiologic abnormalities are seen in a majority of patients with lymphangioleiomyomatosis. On ventilation scintigrams, a frequently seen speckling pattern may be related to accumulation of radionuclide in pulmonary cysts-a hallmark of the disease at CT. Findings with each imaging modality correlate with certain pulmonary functions.

Markers of cell proliferation and expression of melanosomal antigen in lymphangioleiomyomatosis.

Pulmonary lymphangioleiomyomatosis (LAM), a disease of young women, is characterized by proliferation of immature-appearing smooth-muscle cells (LAM cells) in the lungs and abdomen. LAM cells react with monoclonal antibody HMB45, which recognizes a 100-kD glycoprotein (gp100) originally found in human melanoma cells. We investigated the expression and the subcellular localization of gp100 in lung tissue from patients with LAM and in human melanoma cell lines (Malme-3M, A2058, and CHL-1), and the relationship between this expression and cellular proliferation. Binding sites for HMB45 antibody in melanoma and LAM cells were located in cytoplasmic granules resembling immature melanosomes. LAM cells reactive for proliferating-cell nuclear antigen (PCNA), a marker of cellular proliferation, were spindle-shaped, in contrast to the large, epithelioid cells reacting with HMB45 antibody. In accord with this finding, we observed an inverse relationship between the immunostaining for HMB45 antibody and PCNA in LAM and melanoma cells. Thus, LAM and melanoma cells are heterogeneous with respect to their stages of proliferation and their expression of melanoma antigens. PCNA-positive cells, which are more likely to be negative for reactivity with HMB45 antibody, may be more relevant to the progression of LAM than are HMB45-positive cells, which are the hallmark of LAM.

Fluctuation of serum leptin level in rats after ovariectomy and the influence of estrogen supplement.

In order to understand the mechanism of increasing body fat in perimenopausal and postmenopausal women, an ovariectomy-induced obesity model was used to study the role of leptin. In this investigation, a long-term study lasted for 13 weeks was conducted to monitoring the change of serum leptin level in rats after the loss of estrogen, and also to examine the influence of estrogen replacement. The results showed that three weeks after the removal of ovaries the body weight of Ovx rats was already significantly higher than the other two groups, and continued to gain more weight thereafter. Accompanying with the significant weight gain was the changes in the serum leptin levels. The leptin concentration declined gradually during the first half of experimental period, dropping down to an almost undetectable level at week 7 (0.216+/-0.132 ng/ml). Subsequently, its concentration began to elevate, and by the end of experiment leptin level was significantly higher (3.182+/-0.936 ng/ml) than the value before the operation (0.818+/-0.242 ng/ml). This fluctuation of serum leptin level caused by ovariectomy was eliminated by the replacement of estrogen. The present data indicate that ovariectomy-induced weight gain is caused by the early drop in leptin level. The later rise in leptin production is connected to the increased body weight probably originated from a reduced sensitivity in leptin signal.

Comprehensive evaluation of 35 patients with lymphangioleiomyomatosis.

OBJECTIVES: To evaluate comprehensively the characteristics of lymphangioleiomyomatosis (LAM), with emphasis on the application of imaging and immunohistochemical methods. DESIGN: Prospective study. PATIENTS: Thirty-five female subjects with LAM. SETTING: Clinical Center, National Institutes of Health. INTERVENTIONS: BAL, pulmonary function test, ventilation/perfusion lung scans, CT of the chest and abdomen, ultrasonography of abdomen, and immunohistochemical study of lung biopsy specimens. RESULTS: Most patients had exertional dyspnea (83%) and pneumothorax (69%). BAL did not show diagnostic changes. The most common abnormalities on pulmonary function tests were decreased diffusing capacity of carbon monoxide (83%), hypoxemia (57%), and airway obstruction (51%). Bronchodilator response was found in 26% of patients. CT, which is almost pathognomonic, showed numerous thin-walled cysts throughout both lungs in all patients. Thirty-four patients (97%) had abnormal ventilation and/or perfusion lung scans. An unusual "speckling" pattern was observed on ventilation scans of 74% of patients. Common extrapulmonary features were retroperitoneal adenopathy (77%) and renal angiomyolipomas (60%). The percentage of abnormal smooth muscle cells (LAM cells), reactive with HMB45, varied from 17 to 67% in 10 lung biopsy specimens. CONCLUSIONS: Improved diagnostic methods have defined the abnormalities in patients with pulmonary LAM and increased the potential for early recognition and treatment of this disorder. Patients with LAM should be evaluated for bronchodilator responsiveness and may benefit from a trial of bronchodilators.

Immunohistochemical analysis of proteins of the Bcl-2 family in pulmonary lymphangioleiomyomatosis: association of Bcl-2 expression with hormone receptor status.

BACKGROUND: Pulmonary lymphangioleiomyomatosis (LAM), a disease of young women, is characterized by proliferation of abnormal smooth muscle cells (LAM cells), which often differ from normal pulmonary smooth muscle cells by frequently having estrogen and progesterone receptors. OBJECTIVE: To evaluate the relationships among several factors related to proliferation and apoptosis in LAM cells, we employed immunohistochemical methods for the localization of Bcl-2 and Mcl-1 (inhibitors of apoptosis), Bax (a promoter of apoptosis), c-Myc (an apoptosis-related oncoprotein), proliferating cell nuclear antigen (an indicator of mitotic activity), and nick end labeling (to identify apoptotic cells) in lung tissues of 9 patients with LAM. RESULTS: In all patients, most LAM cells reacted positively for Bax. The LAM cells were positive for both Bcl-2 and estrogen receptor in 5 patients, positive for only Bcl-2 in 1 patient, positive for only estrogen receptor in another patient, and negative for both in 2 patients. More than 50% of the Bcl-2-positive LAM cells were also positive for estrogen receptor. The reaction for c-Myc was positive in all patients. The immunoreactivity for Bcl-2 and Mcl-1, which inhibit apoptosis, was more intense in LAM cells than in normal vascular and bronchial smooth muscle cells. In 6 patients, more than 50% of the LAM cells were positive for proliferating cell nuclear antigen. Apoptosis was infrequent in LAM cells. CONCLUSIONS: Our results suggest that the expression of Bcl-2 in LAM cells may be related to hormonal regulation, and that by decreasing apoptosis, Bcl-2 and related proteins contribute to the imbalance between proliferation and death of LAM cells.

Cytokine-mediated transcriptional induction of the human inducible nitric oxide synthase gene requires both activator protein 1 and nuclear factor kappaB-binding sites.

The involvement of AP-1 and NF-kappaB transcription factors in cytokine-mediated induction of human inducible nitric oxide synthase (hiNOS) promoter activity was examined. Luciferase reporter plasmids, containing mutations in AP-1 and NF-kappaB sites, in a hiNOS promoter extending from -8.3 kilobase pairs (kb) to +168, were transiently expressed in A549 cells, and promoter activity was determined after treatment with a cytokine mixture (CM) containing interleukin 1-beta, interferon-gamma, and tumor necrosis factor-alpha. Mutation of the AP-1 heptad located -5301 base pairs upstream decreased gene activation by 90% in a -8.3-kb promoter and a shorter -5.574-kb promoter. Disruption of AP-1 (at -5115) or NF-kappaB (at -115 and -8283) sites reduced promoter activity by 45, 67, and 52%, respectively. Responsiveness to CM was decreased by 85% in constructs mutated in both NF-kappaB sites. By gel retardation analyses, CM increased AP-1- and NF-kappaB binding. Supershift analysis identified Jun D and Fra-2 as components of AP-1 complexes. Each kappaB site bound different complements of NF-kappaB/Rel family members (downstream site, Rel A/p50; upstream site, Rel A/Rel A). Rel A was maximally, whereas IkappaB-alpha was minimally, expressed in nuclei after 1 h of CM treatment, corresponding with the peak in NF-kappaB inding activity. Thus, AP-1 and NF-kappaB are important cis-elements for induction of hiNOS gene transcription.

Analysis of the cytokine-stimulated human inducible nitric oxide synthase (iNOS) gene: characterization of differences between human and mouse iNOS promoters.

Expression of human inducible nitric oxide synthase (hiNOS) is under cytokine control and is transcriptionally regulated. The hiNOS and mouse iNOS (miNOS) genes are regulated differently by cytokines. To understand better the transcriptional regulation of the hiNOS gene, the 8.3-kb hiNOS promoter was characterized. Promoter activity was evaluated by transient transfection of hiNOS luciferase constructs in A549 human alveolar type II epithelium-like cells in the presence and absence of cytokines (IFN-gamma, IL-1 beta, and TNF-alpha). Important cytokine-responsive elements are located at -3665 to -5574 bp (containing two perfectly matched AP-1 sites which are not present in miNOS promoter) and -8093 to -8296 bp (one perfectly matched NF-kappa B site) of the hiNOS promoter region. Likely, these two AP-1 sites and the upstream NF-kappa B site are important in the transcriptional induction of hiNOS by cytokines. Our data demonstrate the molecular basis for the different cytokine-stimulated characteristics of hiNOS and miNOS genes.

Proteolytic activities of mouse sarcoma 180 cells that are inhibited by Bowman-Birk and Kunitz protease inhibitors.

In this study, using zymogram analysis two proteolytic activities were identified in the mouse sarcoma 180 (S-180) cells that were activated by trypsin treatment and inhibited by both BBI and ACTI. These enzymes, with molecular weights of 46 kDa (dominant band) and 62 kDa (minor band), were mainly localized in the cytosol, and had optimal activity at pH 7 and 8 respectively. Their inhibition by DFP, BBI and ACTI but not EDTA and TPCK indicated they were trypsin-like serine proteases and may be the intracellular target-enzymes of protease inhibitors. The level of the precursor of the 62 kDa protease was significantly increased in the S-180 solid and soft tumors, whereas the level of the 46 kDa precursor was almost undetectable, implying that a physiological role may be played by these serine proteases during tumor invasion.

Database issues in object-oriented clinical information systems design.

A clinical information system (CIS) prototype was created from an Object-Oriented (OO) design. We experienced considerable difficulties when implementing the OO data model in a relational database management system (RDBMS), including lack of semantic power and support for complex objects, inability to encapsulate object methods, and performance degradation due to extensive join operations. This paper reflects on the experiences of a CIS research project and explores issues related to the use of RDBMS and Object-Oriented Database Management Systems (OODBMS) in CIS design and development.