Human pleural fluid is a potent growth medium for Streptococcus pneumoniae.

Empyema is defined by the presence of bacteria and/or pus in pleural effusions. However, the biology of bacteria within human pleural fluid has not been studied. Streptococcus pneumoniae is the most common cause of pediatric and frequent cause of adult empyema. We investigated whether S. pneumoniae can proliferate within human pleural fluid and if growth is affected by the cellular content of the fluid and/or characteristics of pneumococcal surface proteins. Invasive S. pneumoniae isolates (n = 24) and reference strain recovered from human blood or empyema were inoculated (1.5×106CFU/mL) into sterile human malignant pleural fluid samples (n = 11). All S. pneumoniae (n = 25) strains proliferated rapidly, increasing by a median of 3009 (IQR 1063-9846) from baseline at 24hrs in all pleural effusions tested. Proliferation was greater than in commercial pneumococcal culture media and concentrations were maintained for 48hrs without autolysis. A similar magnitude of proliferation was observed in pleural fluid before and after removal of its cellular content, p = 0.728. S. pneumoniae (D39 strain) wild-type, and derivatives (n = 12), each with mutation(s) in a different gene required for full virulence were inoculated into human pleural fluid (n = 8). S. pneumoniae with pneumococcal surface antigen A (ΔpsaA) mutation failed to grow (2207-fold lower than wild-type), p<0.001, however growth was restored with manganese supplementation. Growth of other common respiratory pathogens (n = 14) across pleural fluid samples (n = 7) was variable and inconsistent, with some strains failing to grow. We establish for the first time that pleural fluid is a potent growth medium for S. pneumoniae and proliferation is dependent on the PsaA surface protein and manganese.

Protocol of the Australasian Malignant Pleural Effusion-2 (AMPLE-2) trial: a multicentre randomised study of aggressive versus symptom-guided drainage via indwelling pleural catheters.

INTRODUCTION: Malignant pleural effusions (MPEs) can complicate most cancers, causing dyspnoea and impairing quality of life (QoL). Indwelling pleural catheters (IPCs) are a novel management approach allowing ambulatory fluid drainage and are increasingly used as an alternative to pleurodesis. IPC drainage approaches vary greatly between centres. Some advocate aggressive (usually daily) removal of fluid to provide best symptom control and chance of spontaneous pleurodesis. Daily drainages however demand considerably more resources and may increase risks of complications. Others believe that MPE care is palliative and drainage should be performed only when patients become symptomatic (often weekly to monthly). Identifying the best drainage approach will optimise patient care and healthcare resource utilisation. METHODS AND ANALYSIS: A multicentre, open-label randomised trial. Patients with MPE will be randomised 1:1 to daily or symptom-guided drainage regimes after IPC insertion. Patient allocation to groups will be stratified for the cancer type (mesothelioma vs others), performance status (Eastern Cooperative Oncology Group status 0-1 vs ≥2), presence of trapped lung (vs not) and prior pleurodesis (vs not). The primary outcome is the mean daily dyspnoea score, measured by a 100 mm visual analogue scale (VAS) over the first 60 days. Secondary outcomes include benefits on physical activity levels, rate of spontaneous pleurodesis, complications, hospital admission days, healthcare costs and QoL measures. Enrolment of 86 participants will detect a mean difference of VAS score of 14 mm between the treatment arms (5% significance, 90% power) assuming a common between-group SD of 18.9 mm and a 10% lost to follow-up rate. ETHICS AND DISSEMINATION: The Sir Charles Gairdner Group Human Research Ethics Committee has approved the study (number 2015-043). Results will be published in peer-reviewed journals and presented at scientific meetings. TRIAL REGISTRATION NUMBER: ACTRN12615000963527; Pre-results.

ADAM17_i33708A>G polymorphism interacts with dietary n-6 polyunsaturated fatty acids to modulate obesity risk in the Genetics of Lipid Lowering Drugs and Diet Network study.

BACKGROUND AND AIMS: The disintegrin and metalloproteinase ADAM17, also known as tumor necrosis factor alpha converting enzyme, is expressed in adipocytes. Importantly, elevated levels of ADAM17 expression have been linked to obesity and insulin resistance. Therefore, the aim of this study was to evaluate the association of six ADAM17 single nucleotide polymorphisms (SNPs) (m1254A>G, i14121C>A, i33708A>G, i48827A>C, i53440C>T, and i62781G>T) with insulin-resistance phenotypes and obesity risk, and their potential interactions with dietary polyunsaturated fatty acids (PUFA). METHODS AND RESULTS: ADAM17 SNPs were genotyped in 936 subjects (448 men/488 women) who participated in the Genetics of Lipid Lowering Drugs and Diet Network (GOLDN) study. Anthropometrical and biochemical measurements were determined by standard procedures. PUFA intake was estimated using a validated questionnaire. G allele carriers at the ADAM17_m1254A>G polymorphism exhibited significantly higher risk of obesity (P=0.003), were shorter (P=0.017), had higher insulin (P=0.016), and lower HDL-C concentrations (P=0.027) than AA subjects. For the ADAM17_i33708A>G SNP, homozygotes for the A allele displayed higher risk of obesity (P=0.001), were heavier (P=0.011), had higher BMI (P=0.005), and higher waist measurements (P=0.023) than GG subjects. A significant gene-diet interaction was found (P=0.030), in which the deleterious association of the i33708A allele with obesity was observed in subjects with low intakes from (n-6) PUFA (P<0.001), whereas no differences in obesity risk were seen among subjects with high (n-6) PUFA intake (P>0.5) CONCLUSION: These findings support that ADAM17 (m1254A>G and i33708A>G) SNPs may contribute to obesity risk. For the ADAM17_i33708A>G SNP, this risk may be further modulated by (n-6) PUFA intake.

Pulmonary tumour with high carcinoembryonic antigen titre caused by chronic propolis aspiration.

Carcinoembryonic antigen (CEA) titre elevation is sometimes found in benign diseases, such as gastro-intestinal tract inflammatory disease and chronic obstructive pulmonary disease; however, very high CEA titre is rarely encountered in benign pulmonary disease. A 36-yr-old female, who had suffered from body weight loss, was found to have high serum CEA titre (60.8 ng.mL(-1)). Image studies revealed one pulmonary tumour at the left lower lobe, satellite nodules and mediastinal lymphadenopathy. Left lower lobectomy and lymph node dissection were performed for suspicious pulmonary malignancy. The pathological examination revealed that the tumourous lesion was composed of small and fragmented foreign bodies, fibrinopurulent exudate and heavy eosinophils. The bronchial epithelium was characterised by goblet cell hyperplasia and CEA overexpression. The remaining lung parenchyma possessed similar foreign body reaction. The patient's medical history was reviewed and it was found that she had spread propolis topically on nasal mucosa as an adjuvant therapy to asthma for 6 months prior to this medical event. The CEA titre decreased after the operation to 14.2 and 7.88 ng.mL(-1) after 2 weeks and 6 months, respectively. Propolis is used widely in folk medicine but it also has strong sensitising potential. One rare case of propolis aspiration is reported with presentation mimicking lung cancer.

Differential regulation of protein kinase C activity by modulating factors and Euonymus alatus (Thunb.) Sieb in human myometrial and uterine leiomyomal smooth muscle cells.

Human myometrial smooth muscle cells (MSMC) showed high protein kinase C (PKC) activity when a maximal dose of PKC-activating phorbol ester was used, while uterine leiomyomal cells (ULMC) showed only 6-12% of PKC activity. MSMC exhibited a low proliferation rate, whereas ULMC exhibited a high proliferation rate. These different cell types of MSMC and ULMC responded to 10 U/mL thrombin, with a twofold stimulation of PKC activity. Downregulation of PKC activity was found when MSMC were treated with phorbol ester or with transforming growth factor-beta2. We concluded that differences in PKC activity exist between MSMC and ULMC, which may be related to their different proliferative activity. ULMC treated with Euonymus alatus (Thunb.) Sieb (EA), known as "gui-jun woo" in Korea, which is used for leiomyomal tumors, exhibited a much lower proliferation rate than untreated cells, suggesting that EA inhibited the cellular proliferation of ULMC. The PKC activity of MSMC by EA treatment (50 microg/mL) changed little. ULMC showed increased PKC activity by addition of EA, indicating that PKC is activated by EA. The EA-treated ULMC were differentiated into phenotypes characteristic for normal untransformed cells, since the EA-treated cells possess higher PKC activity than untreated cells.

Vermicomposting of cattle and goat manures by Eisenia foetida and their growth and reproduction performance.

Vermicomposting is commonly adopted for the treatment of livestock organic wastes. In the present study, two types of livestock manure were used for culturing of the earthworm, Eisenia foetida. Each treatment group consisted of six replicates and worm vermicasts were examined after 5 weeks. The concentrations of total C, P and K in goat manure vermicasts were higher than those in cattle manure vermicasts. Cattle vermicasts had a higher N content than goat vermicasts but the C:N ratio of fresh manure was higher than that of vermicasts for both materials. Earthworm biomass and reproductive performance, in terms of number of worms after 5 weeks of experiment, were higher in cattle manure than in goat manure. The cocoon production per worm in cattle manure was higher than in goat manure. However, the hatchability of cocoons was not affected by manure treatments. In conclusion, cattle manure provided a more nutritious and friendly environment to the earthworms than goat manure.

Scutellaria barbata D. Don induces c-fos gene expression in human uterine leiomyomal cells by activating beta2-adrenergic receptors.

Scutellaria barbata D. Don (Lamiaceae; SB) inhibited the growth of uterine leiomyomal (LM) cells with unknown actions. The expression patterns of beta-adrenergic receptors (beta-ARs) in human uterine LM cells and functional coupling to gene expression have also been investigated. Northern blot analysis showed that beta-AR subtypes are expressed at different levels in the uterine LM cells and myometrial smooth muscle cells (SMCs). beta1-AR expression was to be found approximately at the same level in the two cell types. beta2-ARs were expressed at higher levels in uterine LM cells than that in myometrial SMCs. beta3-AR expression was not found in both the cells. c-fos gene expression was induced by SB in uterine LM cells via increases in adenosine-3',5', cyclic monophosphate (cAMP), which in turn activated the cAMP/protein kinase A (PKA) pathway. The PKA inhibitor, H89, inhibited c-fos gene expression induced by SB. It seems that the mechanism of proto-oncogenes c-fos different leiomyoma from other myometrial cancer. Further studies are necessary to elucidate whether c-fos induction by SB in uterine LM cells influences a regression of leiomyoma or induces other differentiation.

Transforming growth factor beta induces vascular endothelial growth factor elaboration from pleural mesothelial cells in vivo and in vitro.

Vascular endothelial growth factor (VEGF) increases vascular permeability and is important in pleural effusion formation. In studies using transforming growth factor beta (TGF-beta) to produce pleurodesis, we observed that although TGF-beta was more effective than talc or doxycycline, it induced transient production of large pleural effusions. We hypothesized that TGF-beta stimulates VEGF production in pleural tissues in vivo, and by mesothelial cells in vitro. New Zealand White rabbits (n = 33) were given TGF-beta(2) (1.7 or 5.0 microg), talc (400 mg/kg), doxycycline (10 mg/kg), or buffer intrapleurally. Pleural fluid was collected at 24 h. Intrapleural injection of TGF-beta(2) induced a dose-dependent increase in VEGF production. The pleural fluid VEGF level was 2-fold higher in rabbits given 5.0 microg of TGF-beta(2) than in those given 1.7 microg of TGF-beta(2) and 3-fold higher than in those given buffer. VEGF levels were higher after the injection of TGF-beta(2) than after administration of talc or doxycycline. The pleural fluid VEGF correlated significantly with the volume of pleural effusions (r = 0.79, p < 0.00001). In vitro, TGF-beta(2) stimulated a dose-dependent increase in VEGF production from murine pleural mesothelial cells. At 4 and 24 h, TGF-beta(2), but not talc or doxycycline, induced a significant increase in VEGF, when compared with controls. The mesothelial cell VEGF production was significantly reduced by anti-TGF-beta antibody in the TGF-beta(2), talc, and control (buffer and medium) groups. In conclusion, mesothelial cells are an important source of VEGF. TGF-beta increases the VEGF production by mesothelial cells in vivo and in vitro.

Risk of betel chewing for oesophageal cancer in Taiwan.

Among 104 cases of squamous-cell oesophageal carcinoma patients and 277 controls in Taiwan, after adjusting for cigarette smoking, alcohol consumption, and other confounders, we found that subjects who chewed from 1 to 495 betel-year and more than 495 betel-years (about 20 betel quid per day for 20 years) had 3.6-fold (95% Cl = 1.3-10.1) and 9.2-fold risk (95% Cl = 1.8-46.7), respectively, of developing oesophageal cancer, compared to those who did not chew betel.

Chemical synthesis of a P(k)-antigenic globotriose analogue with a functionalized aglycon.

6-Aminohexyl alpha-D-galactopyranosyl-(1-->4)-beta-D-galactopyranosyl-(1-->4)-beta-D-glucopyranoside (2), a globotriose analogue with a functionalized aglycon, was synthesized, using alpha-D-galactopyranosyluronic acid-(1-->4)-D-galactopyranosyluronic acid [di-GalA (3)] as the starting material, which is commercially available or can be readily prepared from pectin.

Measurements of dispersion curves of leaky Lamb waves using a lensless line-focus transducer.

This paper presents a new method of measuring dispersion curves of leaky Lamb waves propagating in a thin plate. The measurement system is based on a lens-less line-focus transducer and its defocusing measurement technique. A new waveform processing method known as V(f, z) method is introduced for analyzing the measured waveforms and for extracting the dispersion relation. For two thin-plate samples, namely a 150 microm thick alumina plate and a 320 microm thick stainless steel plate, the dispersion curves of several lower order modes of leaky Lamb waves have been experimentally determined in the frequency range of few to 25 MHz. The experimental dispersion curves have then been compared with the theoretical ones. Excellent agreement is observed. It is then verify that this method indeed provides a convenient and accurate way for experimentally measuring dispersion curves of leaky Lamb waves of thin-plate samples.

Genetic polymorphisms of XRCC1 and risk of the esophageal cancer.

A variety of environmental factors were identified to be associated with the risk of esophageal cancer. The variation in capacity of DNA repair might influence environmental chemical-associated carcinogenesis. We hypothesized that the polymorphic XRCC1 genes might modify cancer susceptibility of the esophagus. To investigate the effect of XRCC1 genetic polymorphisms on codons 194, 280 and 399, we evaluated data from 105 patients of esophageal squamous cell carcinoma and 264 healthy controls, matching with age (+/-3 years), gender and ethnicity. The distribution of the 3 genotypes were not significantly different among patients and controls. However, among alcohol drinkers, the XRCC1399 Arg/Arg genotype was more frequently found in patients with esophageal cancer. After adjustment with other environmental confounders, the OR for the genotype of XRCC1399 Arg/Arg was 2.78 (95% CI =1.15-6.67) as compared with the XRCC1(399) Arg/Gln and XRCC1(399) Gln/Gln genotypes in the alcohol drinkers. Similar trends were observed among cigarette smokers and areca chewers. However, they did not reach a statistical significance. Our findings suggest that the polymorphic XRCC1 genes might modify the risk of alcohol-associated esophageal cancers.

Effect of Jindangwon on streptozotocin-induced diabetes.

The inhibitory effects of the traditional herbal medicine Jindangwon (JDW) on streptozotocin (ST)-induced diabetic mellitus were studied using the ST-treated diabetic model. Glucokinase activity of pancreatic islets was severely impaired by ST treatment. However, when ST-treated islets were treated with 1 mg/ml of JDW, the enzyme activities of glucokinase and hexokinase were protected, glucose-6-phosphatase was not. When the effects of JDW on ST-induced ATP/ADP ratio of islets were assayed, JDW was effective in restoring of ATP/ADP ratio. In addition, ST decreased the enzyme activities of PDH, while JDW had a protective effect on the enzyme. ST-induced cGMP accumulation was significantly inhibited by JDW treatment. Furthermore, ST-induced nitrite formation was significantly inhibited by JDW treatment. JDW also showed the suppressed nitrite production in ST-treated pancreatic islet cells. When the islets (200/condition) were treated with ST (5 mM for 30 min), and then JDW was added to the ST-treated cells, 1.0 mg/ml of JDW showed the activated and recovered aconitase activity in pancreatic islet cells. When the effect of ST on the gene expression of pancreatic GLUT2 and glucokinase were examined, the level of GLUT2 and glucokinase mRNA in pancreatic islets was significantly decreased. However, JDW protected and improved the expression of protein and genes, indicating that JDW is effective on ST-induced inhibition of gene expression of GLUT2, glucokinase and proinsulin in islets. These results suggested that JDW is effective in this model to treat ST-induced diabetes.

Activating signal cointegrator 1, a novel transcription coactivator of nuclear receptors, and its cytosolic localization under conditions of serum deprivation.

Activating signal cointegrator 1 (ASC-1) harbors an autonomous transactivation domain that contains a putative zinc finger motif which provides binding sites for basal transcription factors TBP and TFIIA, transcription integrators steroid receptor coactivator 1 (SRC-1) and CBP-p300, and nuclear receptors, as demonstrated by the glutathione S-transferase pull-down assays and the yeast two-hybrid tests. The ASC-1 binding sites involve the hinge domain but not the C-terminal AF2 core domain of nuclear receptors. Nonetheless, ASC-1 appears to require the AF2-dependent factors to function (i.e., CBP-p300 and SRC-1), as suggested by the ability of ASC-1 to coactivate nuclear receptors, either alone or in cooperation with SRC-1 and p300, as well as its inability to coactivate a mutant receptor lacking the AF2 core domain. By using indirect immunofluorescence, we further show that ASC-1, a nuclear protein, is localized to the cytoplasm under conditions of serum deprivation but is retained in the nucleus when it is serum starved in the presence of ligand or coexpressed CBP or SRC-1. These results suggest that ASC-1 is a novel coactivator molecule of nuclear receptors which functions in conjunction with CBP-p300 and SRC-1 and may play an important role in establishing distinct coactivator complexes under different cellular conditions.

Segmental necrosis of small bronchi after prolonged intakes of Sauropus androgynus in Taiwan.

Since 1994 an endemic of chronic obstructive pulmonary disease (COPD) has developed in Taiwan after a prevalent use of a body-weight-reducing vegetable, Sauropus androgynus (SA). All conventional treatments for COPD, including steroids and bronchodilators, had been ineffective. Studies of limited lung biopsy specimens from these patients revealed bronchiolitis obliterans. A few patients died, but many developed protracted chronic respiratory failure. Because of the chronic debilitation and ineffective conventional treatments, single lung transplants were performed as the last resort in four patients. The excised lungs revealed focal fibromuscular sclerosis and obliteration of bronchial arteries in the wall of large bronchi 4 to 5 mm in diameter with segmental necrosis of bronchi 2 to 4 mm in diameter. Bronchi immediately proximal to the necrotic zone showed fibrosis and atrophy of cartilage, bronchial glands, and smooth muscle cells; bronchioles immediately distal showed obstruction or dilatation. Most bronchi larger than 5 mm, pulmonary vessels, small bronchioles, and alveoli were little altered. The pathologic changes were most consistent with segmental ischemic necrosis of bronchi at the water-shed zone of bronchial and pulmonary circulation. The specific etiologic agent and detail of pathogenesis of this SA-related COPD needs further investigation.

Role of nitric oxide in penile erection.

The present study was undertaken to investigate the role of nitric oxide (NO) in erectile physiology by correlating its action with the existence and activity of nitric oxide synthase (NOS), which produces NO. We applied Western blot analysis in both human and rat penile tissue. In the rat, reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining and spectrophotometric assay were also performed, in addition to in vivo electroerection study with pharmacological manipulation. Western blot analysis identified a protein of 155 KDa identical to the neural form of NOS in the human and rat penis. The NOS blot densities in the two species were similar, and both were lower than that in the rat cerebellum. Histochemical staining localized NOS to neurons innervating the corpora cavernosa, including the pelvic plexus, the cavernosal nerves and their terminal fibers within the corporeal erectile tissue, and dorsal penile nerves. NOS activity was also found in the cerebellum, urethra, penis, and urinary bladder, in decreasing order of intensity. Intracavernous injections of NOS inhibitor (L-NOARG or L-NAME in concentrations from 10(-6) M to 10(-3) M suppressed electrostimulation-induced erection in a concentration-dependent manner. Subsequent intracavernous injection of L-Arginine (10(-2) M) partially restored the erection. The neural form of constitutive NOS in the corpora cavernosa synthesizes NO, which mediates penile erection. Determination of cavernosal NOS expression or activity may permit characterization of certain pathological conditions that cause impotence.

Use of the silicone tracheal T-tube for tracheostenosis or tracheomalacia.

BACKGROUND: Tracheobtenosib and tracheomalacia are trivial diseases. The conventional choice of managements with tracheostomy, either temporary or long-term usage, can only partially resolve the problems of airway obstruction. Silicone tracheal T-tube presents a substitute for it. METHODS: We present 5 patients with tracheostenosis or tracheomalacia managed with nine procedures of long silicone Montgomery T-tube prothesis between 1984 and 1994 in VGH-Taipei. The primary diagnosis included tracheal injury (2), postintubation tracheal stenosis (2), and stenosis due to endotracheal tuberculosis (1). Three patients received a long segmental T-tube for permanent endotracheal stenting and the other two patients used T-tube insertion for temporary stenting of the trachea for 7 and 11 months, respectively, with satisfactory results. RESULTS: All patients got immediate benefit from the prothesis in respiration with simple postoperative care. Two patients with temporary T-tube placement had it successfully removed in 7 and 11 months, respectively. Placement of the T-tube for subglottic stenosis also protected the function of phonation. The tracheal T-tube restored airway patency reliably with good long-term results and could be the preferred management of chronic upper airway obstructive disease not amenable to surgical repair. The most common complication was airway obstruction caused by either granulations or sticky mucoid substance. Three patients and six tubes (60%) developed granulation obstruction and the average duration of granuloma formation was 7.7 months. Laser phototherapy or surgical intervention, such as tracheoplasty, with change of the T-tube was carried out for granuloma obstruction. CONCLUSIONS: T-tube is a good endoprothesis for tracheostenosis and tracheomalacia with minimal complication for cases of long tracheostenosis or complex tracheal injury.

Immunologic and physicochemical studies of Bermuda grass pollen antigen BG60.

BACKGROUND: In a previous study we showed that antigen BG60 of Bermuda grass pollen contains isoallergens. Because the yield of purified isoallergens was low when a chromatofocusing technique was used, it was difficult to carry out further studies, such as determination of carbohydrate composition and structure. OBJECTIVE: The aim of this study was to establish a procedure to purify antigen BG60 proteins as a group and to characterize this group's physicochemical and immunologic properties. METHODS: A combination of chromatographic techniques (ion-exchange, gel filtration, blue gel affinity, and reverse-phase high-performance liquid chromatography) was used for the purification of BG60. Immunoblot and ELISA techniques were used to study BG60-specific IgE and IgG antibodies in patients' sera. The role of the carbohydrate moiety in antigenicity and allergenicity was examined with monoclonal antibodies and allergic sera by using periodate-treated BG60. Its carbohydrate composition was analyzed by high-performance anion-exchange chromatography with a pulsed amperometric detector. RESULTS: Homogeneity of BG60 was demonstrated by a single sharp peak in reverse-phase high-performance liquid chromatography, a single band in sodium dodecylsulfate-polyacrylamide gel electrophoresis, and only one band stained by anti-BG60 monoclonal antibody. BG60-specific IgE and IgG antibodies were shown to be present in allergic sera. Six plant lectins were found to react with BG60. On periodate treatment, BG60 reduced binding toward its monoclonal antibody and human IgE and IgG. Carbohydrate composition analysis showed that BG60 contains three kinds of sugars: mannose, N-acetylglucosamine, and fucose (in a ratio of approximately 3:2:1) and a minute amount of xylose. The carbohydrate content is approximately 7.5%, and peptide content is about 92.5%. CONCLUSION: A procedure was established for the purification of a large quantity of the BG60 antigen. The results suggest that the carbohydrate moiety of antigen BG60 may play an important role in the immune response.

Adrenocortical pregnenolone binding activity resides with estrogen sulfotransferase.

Cytosol prepared from Chinese hamster ovary (CHO)-K1 cells transfected with guinea pig estrogen sulfotransferase (EST) cDNA demonstrated high affinity binding activity for pregnenolone as well as 17 beta-estradiol but failed to bind dehydroepiandrosterone or testosterone. In contrast, cytosol prepared from nontransfected CHO-K1 cells did not demonstrate steroid binding activity. Additionally, the binding activity for pregnenolone and 17 beta-estradiol was dependent on the presence of the cofactor adenosine-3',5'-diphosphate. Pregnenolone and 17 beta-estradiol effectively competed with each other for binding. On the other hand, pregnenolone, which was not sulfonated, did not inhibit the sulfonation of 17 beta-estradiol by expressed EST.

Immunostimulation by Alsophila spinulosa extract fraction VII of both humoral and cellular immune responses.

Alsophila spinulosa (Cyatheaceae) has been used in Asia as a herbal medicine. In this study, the immunomodulatory effects of Alsophila spinulosa dried stems water extract fraction VII (AS-VII) were studied in vitro using an animal model. The results showed that AS-VII stimulated splenocyte proliferation in both BALB/c and C3H/HeJ mice. By cell fractionation analysis, the results showed that the mitogenic effect of AS-VII was predominantly on B cell population. The antibody response was also augmented by AS-VII in BALB/c and C3H/HeJ mice. The augmentation of immune responses in C3H/HeJ, the lipopolysaccharide (LPS)-hyporesponsive mouse strain, indicated that the activity was not simply due to the LPS contamination which may be present in AS-VII preparation. Furthermore, AS-VII caused an increase in cytotoxic T cell activity in a mixed lymphocyte reaction culture. These data indicate that AS-VII possesses the capability of augmenting both humoral and cellular immune responses.