Areca nut extracts increased expression of inflammatory cytokines, tumor necrosis factor-alpha, interleukin-1beta, interleukin-6 and interleukin-8, in peripheral blood mononuclear cells.

BACKGROUND AND OBJECTIVE: Cytokines represent a central role in inflammatory tissue destruction and regulate the immune responses that may govern the progression of periodontal diseases. This study investigated the effects of areca nut extracts on the expression of inflammatory cytokines, tumor necrosis factor-alpha, interleukin-1beta, interleukin-6 and interleukin-8 in peripheral blood mononuclear cells. The role of oxidative stress of areca nut extracts was also examined using curcumin. MATERIAL AND METHODS: The expression of cytokines in peripheral blood mononuclear cells treated with extracts of ripe areca nut or extracts of tender areca nut was analyzed using enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction. RESULTS: Both extracts of ripe areca nut (< or = 40 microg/mL) and extracts of tender areca nut significantly enhanced the production of tumor necrosis factor-alpha and interleukin-1beta in peripheral blood mononuclear cells in a dose-dependent and time-dependent manner. The kinetics of mRNA expression of both cytokines was also enhanced by areca nut extracts. The stimulatory effects of areca nut extracts on the secretion of tumor necrosis factor-alpha, interleukin-1beta, interleukin-6 and interleukin-8 and on the mRNA expression of tumor necrosis factor-alpha, interleukin-1beta and interleukin-6 at 4 h of incubation were reduced by curcumin (20-50 microm). However, the level of interleukin-8 transcripts was not affected by curcumin. Moreover, interleukin-1beta induction by extracts of tender areca nut, but not by extracts of ripe areca nut, was weakened by 10 microm curcumin. The inhibitory effects of curcumin may vary with different cytokines and with different areca nut extract treatments. CONCLUSION: The complex cytokine profile induced by areca nut extracts-treated peripheral blood mononuclear cells implied the possibility of enhanced local inflammation and altered immune functions by the areca chewing habit. The inhibitory effects of curcumin on cytokine expression suggested that oxidative stress might be involved in areca nut extracts-associated immune alteration.

Protein markers in the microcyst of the posteroventral cochlear nucleus of the gerbil.

Microcysts are most evident in the posteroventral and anteroventral cochlear nuclei (PVCN and AVCN) of the Mongolian gerbil. The origin and contents of the microcyst are not elucidated at present. The present study investigated the possible inclusions in the microcyst by employing immunocytochemical labeling to localize the existence of various protein markers. Thirty and 100 microm thick sections were used to substitute and reconstruct between 6 and 20 paraffin serial sections, respectively. In 30-microm-thick slice sections, immunoreactivity of glial fibrillary acidic protein (GFAP-IR), mitochondria inner membrane (MCA-In-IR), S-100 (S-100-IR), serotonin (5-HT-IR), myelin proteolipid protein (PLP-IR) and substance P (SP-IR) abutted on the perimeter of the microcyst. The immunolabeled SP-positive cells were adjacent to the evagination of the microcyst. In 100-microm-thick slice sections, immunoreactivity of nitric oxide synthase (NOS-IR) and somatostatin (SOM-IR) mainly precipitated as flocculent structures in the small to medium-sized microcysts. 5-HT-IR also precipitated as an elongated flocculent stalk adjacent to the large microcyst or randomly distributed in the neuropil. The findings suggest that GFAP, MCA-In, S-100, 5-HT, PLP, SP, NOS and SOM may be involved in modulating the physiological functions and maintaining micro-environmental homeostasis of the microcyst in the cochlear nucleus of the gerbil.

Perceptions of dyspnea and helpful interventions during the advanced stage of lung cancer: Chinese patients' perspectives.

Dyspnea is a distressful but neglected symptom in oncology practice and research. The aim of this study was to describe the experience of dyspnea and helpful interventions in Chinese patients with advanced lung cancer admitted in the palliative care unit in 1 region in Hong Kong. A qualitative description approach using in-depth interviews was used to guide this study. Eleven participants agreed to be interviewed with age ranging from 51 to 80 years. They have been diagnosed with lung cancer from 1 to 12 months, and all required oxygen therapy from dyspnea. The results of content analysis revealed 4 main themes: (1) characteristics of dyspnea, (2) impact of dyspnea, (3) strategies used to manage dyspnea, and (4) nurses' role in managing dyspnea. Patients in this study found no Chinese words to adequately define and describe dyspnea and relied on sensations they experienced during the dyspnea episode. The impact of dyspnea was multidimensional, and patients used various strategies to manage dyspnea, including avoiding triggers and utilizing traditional Chinese medicine. Healthcare professionals were perceived to play a very inadequate role in assisting patients with dyspnea, and participants suggested that they should take a more active role in educating and supporting patients with dyspnea.

Areca nut extracts reduce the intracellular reactive oxygen species and release of myeloperoxidase by human polymorphonuclear leukocytes.

BACKGROUND AND OBJECTIVE: Polymorphonuclear leukocytes (PMN) represent the first line of host defense. Areca nut extract inhibits the bactericidal activity of, and the release of superoxide anion (O2- ) by, PMN. This study investigated the effects of areca nut extract on the intracellular production of reactive oxygen species (ROS) and on the extracellular release of lysosomal enzyme, myeloperoxidase (MPO), by PMN. The effects of arecoline, a principal component of areca nut, were also examined. MATERIAL AND METHODS: Human PMN were treated with various concentrations of areca nut extract or arecoline followed by treatment with Hanks' balanced salt solution, with or without cytochalasin B and fMet-Leu-Phe (CB/fMLP). The viability of PMN was determined using propidium iodide staining and flow cytometry. The presence of intracellular ROS was determined using 2',7'-dichlorofluorescin diacetate and fluorometry. MPO release was determined using a substrate assay. RESULTS: Areca nut extract (25 and 50 microg/ml) significantly decreased the viability of PMN. The intracellular levels of ROS and the extracellular release of MPO were induced in PMN by CB/fMLP. Exposure of PMN to areca nut extract (up to 25 microg/ml) or to arecoline (up to 2 mg/ml) did not directly affect the levels of ROS and MPO activity. However, under conditions that did not affect the viability of PMN, the ability of CB/fMLP to trigger production of intracellular ROS and release of MPO in human PMN was significantly suppressed by areca nut extract and arecoline. CONCLUSION: Areca nut impaired the activation of PMN by CB/fMLP that might decrease the effectiveness of PMN in the host defense. Alternatively, exposure of PMN to areca nut extract could decrease the capacity of PMN to damage tissues.

Cytotoxic effects of dental resin liquids on primary gingival fibroblasts and periodontal ligament cells in vitro.

Cytotoxic effects of resin liquids of three in situ relining dental polymers, Alike, Kooliner, and Tokuso Rebase, and their major components, methyl methacrylate (MMA), isobutyl methacrylate (IBMA), and 1,6-hexanediol dimethacrylate (1,6-HDMA) were investigated. The concentrations of major monomers in these resin liquids were determined by high-performance liquid chromatography. Cellular viability of human gingival fibroblasts (GF) and periodontal ligament (PDL) cells were evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide assay. Moreover, patterns of cell death were analysed using annexin V/propidium iodide staining with flow cytometry. The results indicated that Alike liquid contained 91.3% MMA, Kooliner liquid contained 94.5% IBMA, and Tokuso Rebase liquid contained 65.8% 1,6-HDMA. All materials examined had cytotoxic effects on GF and PDL cells in dose-dependent manners. Tokuso Rebase liquid appeared to be the most cytotoxic among the various resin liquids examined. The effects of Kooliner and Tokuso Rebase liquids may have resulted from IBMA and 1,6-HDMA, respectively. Furthermore, the majority of treated cells died from necrosis; whereas a small portion of cells died from apoptosis. In conclusion, the results demonstrated that these liquid forms of dental polymers and their major monomers cause cytotoxic reactions. The direct relining procedure that cures these materials in situ should be used cautiously.

Learning and memory in the mosquito Aedes aegypti shown by conditioning against oviposition deterrence.

Oviposition site choice by female mosquitoes, Aedes aegypti (L) (Diptera: Culicidae), was affected by rearing them in water treated with 0.016% of the repellent Mozaway trade mark containing citronella and neem. Given a choice between a bowl of repellent-treated and a bowl of untreated water, Ae. aegypti reared in untreated water strongly preferred to oviposit on the clean water (93-98%) instead of repellent-treated water. This demonstrates effective deterrence of oviposition by dilute Mozaway trade mark. Those reared in repellent-treated water were less deterred: 38-46% of their eggs were laid on the repellent-treated water and 54-62% on the clean water. Evidently the female mosquitoes reared in repellent-treated water were conditioned against oviposition site deterrence, as shown when choice tests were conducted 6 days post-emergence from the treated water. This demonstrates learning and memory in the mosquito Ae. aegypti, with implications for dengue vector surveillance and control.

Neural stem/progenitor cells survive and differentiate better in PD rats than in normal rats.

To investigate the effects of grafted neural stem/mesencephalic progenitor cells (NSCs/MP) on rotational behavior of Parkinson's disease (PD) rats and the influence of intracerebral environment on NSCs/MP, we observed the survival and differentiation of NSCs/MP transplanted into 6-hydroxydopamine (6-OHDA)-lesioned and intact striatums. NSCs/MP were prepared from E(11-15) rats and proliferated in serum-free medium with bFGF for several weeks. One day after being primed with serum/dbcAMP to differentiate, cell suspensions were grafted into 6-OHDA-lesioned and intact striatums respectively. It had been found that NSCs/MP were able to survive better and differentiate into more tyrosine hydroxylase (TH)-positive neurons in 6-OHDA-lesioned striatums than in intact ones, and apomorphine-induced rotations were obviously attenuated in MP graft models. The data suggested that NSCs/MP tend to survive and differentiate into TH-positive neurons in 6-OHDA-lesioned striatums. The data demonstrated that striatums in which DAergic terminals are destroyed by 6-OHDA undergo some changes and thus provide more appropriate conditions for NSCs/MP to differentiate into mature DAergic neurons. Furthermore, the finding that MP had greater relieving effects on rotational behavior than NSCs suggests that NSCs could not be used in clinical therapy of PD unless being induced into MP in vitro before transplantation.

Mesencephalic progenitors can improve rotational behavior and reconstruct nigrostriatal pathway in PD rats.

The aim of this study was to investigate the possibility of mesencephalic progenitors (MP) in treating Parkinson's disease (PD). MP were prepared from E(11-13) rats and proliferated in serum-free medium with basic fibroblast growth factor (bFGF) for 10 days. Cells were then collected and implanted into the striatum only--single grafts or simultaneously into the substantia nigra (SN) and the striatum--double grafts. Twelve weeks after transplantation, DiI, a fluorescent dye, was microinjected into the ipsilateral striatum. Using this strategy, it was found that MP of double grafts had more potent effects on rotational behavior than that of single grafts. Injection of the retrograde tracer DiI into the striatum resulted in fluorescent-labeled cells within the intranigral grafts in double grafts. These data greatly support that MP transplants could not only improve rotational behavior, but does help to re-establish nigrostriatal connections so that it may become one efficient way in treating PD.

Chronic hypoxia attenuates ischemia-reperfusion-induced increase in pulmonary vascular resistance.

This study explored the effect of chronic hypoxia on the elevation of pulmonary vascular resistance caused by ischemia-reperfusion (IR) in anesthetized rats. Experiments were separated into five parts. In Part 1, we examined the increase in left pulmonary vascular resistance (Rpl) after ischemia of the left lung and localized the major site for the increased resistance of the left pulmonary vasculature in both the normoxic and chronic hypoxia groups. Here, IR induced a significant increase in Rpl in the normoxic but not the chronic hypoxia group. This increased Rpl in the normoxic group was attributed to contraction of pulmonary arterial segments. Part 2 and Part 3 were focused on the changes in plasma nitrate/nitrite (NOx) and thromboxane B(2) (TxB(2)) levels. TxB(2) increased significantly in the normoxic group, whereas NOx increased significantly in the chronic hypoxia group, following ischemia. Indomethacin (Part 4) prevented IR-induced increase in Rpl in the normoxic group, whereas the IR-induced increase in Rpl appeared in the chronic hypoxia group after N(G)-nitro-L-arginine methyl ester treatment (Part 5). We conclude that IR elicited increases in the cyclooxygenase products such as TxB(2), which in turn caused an increase in Rpl. However, this increased Rpl was attenuated by elevated NOx in the chronic hypoxia group.

Effect of circumferential undercut placement in the root canal wall on the retention of a tapered post in large and small root canals.

OBJECTIVES: This study examined the influence of root canal space and circumferential undercut placement in the dentine wall on the retention of a prefabricated Root Keeper post. METHODS: Twenty-four extracted single-rooted teeth, having narrow root canal spaces, were decoronated, treated endodontically, and then centrally embedded in resin blocks. All canal spaces were prepared with a no. 2 Peeso reamer to an 8mm depth, and half the specimens were successively treated with a no. 3 Peeso to depths of 5 and 8mm for the small canal group (SC) and large canal group (LC), respectively. An L-type keeper post (Aichi Steel Co.) was luted to each canal with resin cement. The assembly was thermocycled 2000 times (4-60 degrees C), and post retention was measured. Afterwards the dislodged post with the intact structure was sandblasted, and the canal was cleaned and slightly undercut using an inverted cone bur. Six treated posts were recemented for both grooved large canal (GLC) and grooved small canal (GSC) groups, and the repaired retentive strengths were measured. RESULTS: Keeper posts cemented in SC (84.26+/-31.85N) showed better (p<0.05) retention than those cemented in LC (41.92+/-22.36N). Interestingly, the repaired strengths of the recemented post increased in both GSC (104.29+/-11.89N) and GLC (96.65+/-6.91N). All posts, except for one in the GLC, failed at the keeper-post joint. CONCLUSIONS: Prefabricated Root Keeper posts had lower retentive strength in roots with a large canal space. Placement of a circumferential undercut in the root canal wall significantly enhanced the retention of the short tapered posts.

Fullerene derivative attenuates ischemia-reperfusion-induced lung injury.

Reactive oxygen species are the major contributing factors to lung ischemia-reperfusion (IR) injury. In this study, we tested whether a water soluble antioxidant fullerene derivative [C(60)(ONO(2))(7 +/- 2)] attenuates IR lung injury. Young Wistar rats were divided into two groups: control and C(60)(ONO(2))(7 +/- 2). Under ventilation with 95% air-5% CO(2) gas mixture and a 2.5 cm H(2)O end-expiratory pressure, the isolated lungs were perfused with a physiological solution. The experimental protocol included three periods: baseline (10 min), ischemia (45 min) and reperfusion (60 min, ventilated with 95% O(2)-5% CO(2) gas mixture). Before and after ischemia, we measured pulmonary arterial pressure (Ppa), pulmonary venous pressure and lung weight (W). Then, pulmonary capillary pressure and filtration coefficient (K(fc)) were calculated. Ischemia caused increases in Ppa, W and K(fc) in the control group. For most cases, the above ischemia-induced increases were attenuated by the C(60)(ONO(2))(7 +/- 2) pretreatment. Our results suggest that the antioxidant C(60)(ONO(2))(7 +/- 2) attenuates IR-induced lung injury.

Endothelin-1 in citric acid aerosol inhalation-induced airway constriction of guinea pigs.

Endotheline-1 (ET-1) has been shown to enhance tachykinin-induced airway constriction. This study was designed to test whether ET-1 is involved in citric acid-induced bronchoconstriction. Forty-eight anesthetized-paralyzed guinea pigs were divided into six groups of 8 animals each: saline control; citric acid; ET-1; ET-1 + citric acid; BQ123 + ET-1 + citric acid; and BQ788 + ET-1 + citric acid. BQ123 and BQ788 are specific ETA and ETB receptor antagonists, respectively. Each animal in the saline control group received 50 breaths of 4 ml saline aerosol and in all citric acid-treated groups was given 50 breaths of 4 ml aerosol generated from 0.6 M citric acid. In all ET-1-treated groups, each animal was exposed to aerosol generated from 10(-8) M ET-1. The animal in the ET-1 + citric acid group was exposed to ET-1 5 min prior to the citric acid. For the last two groups, each animal was first exposed to aerosol generated from either 10(-5) M BQ123 or 10(-5) M BQ788. Five min later, the animal was exposed to ET-1; and then 5 min later was followed by citric acid. Dynamic respiratory compliance (Crs), forced expiratory volume in 0.1 sec (FEV(0.1)), and maximal expiratory flow at 30% total lung capacity (Vmax 30) were obtained before and 3-15 min after citric acid. Either citric acid or ET-1 inhalation caused significant decreases in Crs, FEV(0.1), and Vmax 30, indicating airway constriction. Citric acid-induced airway constriction, for most cases, was not significantly augmented by ET-1. However, either BQ123 or BQ 788 significantly attenuated the airway constriction induced by the combination of ET-1 and citric acid. Also, in an additional study, either BQ123 or BQ788 significantly attenuated citric acid-induced airway constriction. These data suggest that endogenous ET-1 plays an important role in citric acid aerosol-induced airway constriction in guinea pigs.

Association between betel quid chewing, periodontal status and periodontal pathogens.

The present investigation examined whether an association exists between betel quid chewing and signs of periodontal disease and determined the prevalence of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis by polymerase chain reaction. The periodontal status of 34 betel quid chewers and 32 non-betel quid chewers were compared. A significantly higher prevalence of bleeding on probing was found in betel quid chewers than non-chewers among the subjects with higher plaque level, greater gingival inflammation, deeper probing depth or greater attachment loss. Also, the results suggested that betel quid chewers may harbor higher levels of infection with A. actinomycetemcomitans and P. gingivalis than non-betel quid chewers. The association persists after adjusting for severity of the clinical parameters. In conclusion, betel quid chewing was associated with a higher prevalence of bleeding on probing where higher clinical levels of disease existed, and with a likelihood of subgingival infection with A. actinomycetemcomitans and P. gingivalis.

Appropriateness of antibiotic use in the postpartum period.

OBJECTIVE: To determine the appropriateness of current postpartum antibiotic use in clinical practice. METHODS: Medical records were reviewed for all patients delivering in a 3-month period who received postpartum antibiotics during the delivery hospitalization. Subjects were excluded if they received a single postpartum antibiotic dose as part of a mitral valve prolapse prophylaxis protocol, or if they received no more than one postpartum antibiotic dose for surgical prophylaxis. Characteristics of postpartum antibiotic use were abstracted. RESULTS: Two hundred and eleven of 1537 (14%) delivering patients met the inclusion criteria. Seventy-four (35%) delivered vaginally and 137 (65%) delivered by Cesarean section. Postpartum fevers were found in 40 (54%) of vaginal delivery cases and 80 (58%) of women delivering by Cesarean section who received postpartum antibiotics (p = 0.54). For vaginal deliveries there were no differences in the duration of antibiotic use or number of antibiotic doses based on fever status. For Cesarean deliveries, a fever was associated with more antibiotic doses and a longer duration of antibiotic use. Physician justification for antibiotic use was documented in only 116 cases (55%). CONCLUSIONS: The high proportion of women receiving postpartum antibiotics having no evidence for infection or documented indication for therapy suggests that antibiotics may not be appropriately used in the postpartum period.

Construction of single-chain interleukin-12 DNA plasmid to treat airway hyperresponsiveness in an animal model of asthma.

Allergic asthma is strongly associated with the airway inflammation caused by the dysregulated production of cytokines secreted by the allergen-specific type-2 T helper (Th2) cells. Interleukin (IL)-12 is a heterodimeric cytokine, which strongly promotes the differentiation of naive CD4(+) T cells to the type-1 T helper (Th1) phenotype and suppresses the expression of Th2 cytokines. Therefore, immunotherapy with IL-12 has been suggested as a possible therapy for asthma. In previous studies, we developed a murine model of airway inflammation based on the purified, house dust-mite allergen Der p 1 (Dermatophagodies pteronyssinus) as a clinically relevant allergen. We hypothesized that the expression of IL-12 in the airway may represent an effective therapy for allergic airway diseases. In this study, we investigate whether the local transfer of the IL-12 gene to respiratory tissues modifies allergic inflammation and airway hyper-responsiveness (AHR) in our disease model. To enhance the in vivo delivery of the IL-12 gene, we expressed the murine single-chain IL-12 protein from a nonviral vector to which the two IL-12 subunits (p35 and p40) were linked by a 14- to 18-amino-acid linker. One of these single-chain IL-12s, containing an 18 amino-acid polypeptide linker, was stably expressed and had a high level of biological activity comparable to that of native IL-12 in vitro. In mice with Der p 1-induced asthma, the local administration of this IL-12 fusion gene into the lungs significantly prevented the development of AHR, abrogated airway eosinophilia, and inhibited type-2 cytokine production. These findings indicate that the local transfer of the single-chain IL-12 gene is effective in modulating pulmonary allergic responses and may be a convenient method for future applications of DNA vaccination.

Studies on anthracenes. 3. Synthesis, lipid peroxidation and cytotoxic evaluation of 10-substituted 1,5-dichloro-9(10H)-anthracenone derivatives.

The synthesis of a series of 1,5-dichloro-9(10H)-anthracenones bearing O-linked and N-linked substituents in the 10-position are described. Previous studies have shown that 9-acyloxy 1,5-dichloroanthracenes and 9-acyloxy 1,8-dichloroanthracenes displayed a potential cytotoxic effect. These results have encouraged us in further investigation of potential anthracenone derivatives. Therefore, a series of 10-substituted 1,8-dichloro-9(10H)-anthracenone derivatives were synthesized. These compounds were evaluated for their ability to inhibit the growth of human oral epidermoid carcinoma cells (KB cell line), human cervical carcinoma cells of ME 180 (GBM 8401) and Chinese hamster ovary (CHO) cells, respectively. Compounds 3c and 4c of this series compare favorably in the KB cellular assay with mitoxantrone. Compound 4c showed combined inhibitory action against KB, GBM and CHO cell growth, respectively. In addition, redox property of the compounds for the inhibition of lipid peroxidation in model membranes was determined. Compounds 4b and 4d exhibited stronger antioxidant activity than ascorbic acid, (+)-alpha-tocopherol and mitoxantrone, respectively.

Studies on anthracenes. 2. Synthesis and cytotoxic evaluation of 9-acyloxy 1,8-dichloroanthracene derivatives.

The synthesis and cytotoxic evaluation of 9-acyloxy 1,8-dichloroanthracene derivatives are described. The system selectively reduces the carbonyl group flanked by the peri substituents of the anthracenediones to give the corresponding 1,8-dichloro-9(10H)-anthracenone. Simple acylation of anthracenone occurred with appropriate acyl chlorides in CH2Cl2 with a catalytic amount of pyridine to give the 9-acyloxy-1,8-dichloroanthracene derivatives. Considerable interest has developed in the mechanism of how anthracenones achieve this desirable selectivity. These compounds were evaluated in vitro for their ability to inhibit the growth of human oral epidermoid carcinoma cells (KB cell line), human cervical carcinoma cells of ME 180 (GBM 8401) and Chinese hamster ovary (CHO) cells, respectively, as compared to mitoxantrone. The in vitro cytotoxicity evaluation of 9-acyloxy 1,8-dichloroanthracenes against these above cell lines revealed for most of the compounds a cytotoxic potency lower than that of mitoxantrone. The most active compounds were thus selected for further in vitro biological evaluation and structural modification.

Circumnipple-incision, longitudinal-breast dissection augmentation mammaplasty.

A modification of circumnipple breast augmentation is described, which confines the minimal scar to the base of the nipple. Furthermore, a longitudinal breast parenchyma dissection is performed for breast pocket dissection that limits the injury of the lactiferous ducts and nerves to a minimum, and makes dissection and exposure of the pocket easier, even in patients whose nipples are small. This technique is especially useful in small breasts. We have performed this procedure in 57 patients, and the few complications to date are discussed in this paper.

Breast reconstruction with TRAM flap after subcutaneous mastectomy for injected material (siliconoma).

Many patients throughout the world have had breast augmentation using injectable materials. In the Far East, an adulterated impure silicone has been used by non-medical persons and unlicensed 'cosmetic surgeons' as well as by licensed physicians. The injected material usually causes a granulomatous reaction, tenderness, erythema, pain or discomfort and even skin necrosis. Eventually, the injected breast becomes very hard and develops a distorted contour. Previous treatment was by subcutaneous mastectomy with immediate or delayed reconstruction using a silicone prosthesis. The results of this were often unsatisfactory. We obtained more satisfactory results in 11 patients who had the siliconoma removed by subcutaneous mastectomy and the breast reconstructed with a de-epithelialised TRAM flap.