In Vitro Assessment of 4-Acetyl-Antroquinonol B and Erinacine A in Suppressing Breast Cancer-Induced Osteoclastogenesis.

Bone metastasis in metastatic breast cancer commonly results in osteolytic lesions due to osteoclast activity, promoting bone destruction and tumor progression. The bioactive fungal isolates, 4-acetyl-antroquinonol B (4-AAQB) and erinacine A, have diverse pharmacological and biological activities. However, their effects on breast cancer bone metastasis treatment remain unclear. Our study aimed to examine the impact of 4-AAQB or erinacine A on breast cancer metastases in bone. The effects of 4-AAQB and erinacine A on breast cancer-induced osteoclastogenesis, breast cancer migration, production of prometastatic cytokine (TGF-ß) and marker (MMP-9), as well as potential MAPK signaling transductions were assessed. The results revealed that 4-AAQB and erinacine A effectively suppressed breast cancer-induced osteoclastogenesis and migration, and reduced TGF-ß and MMP-9 production via Erk or JNK signaling transductions, specifically in breast cancer cells or in breast cancer cells-induced osteoclasts. Based on these findings, either 4-AAQB or erinacine A showed promise in preventing breast cancer metastases in bone.

Polysaccharides of Grifola frondosa ameliorate oxidative stress and hypercholesterolaemia in hamsters fed a high-fat, high-cholesterol diet.

OBJECTIVES: This study was to evaluate the antioxidant and anti-hypercholesterolaemia activities of Grifola frondosa in hamsters fed a high-fat, high-cholesterol (HFHC) diet. METHODS: G. frondosa, including fruiting bodies (FGF), fermented mycelia (MGF) and polysaccharides extracted from fruiting bodies (FPS), fermented mycelia (MIP) and fermented broth (BEP) were received intragastrically. Lipid profile and antioxidant status in the blood and liver of hamsters were assessed. KEY FINDINGS: FGF decreased weight gain, serum triglycerides and cholesterol and increased hepatic mRNA expression of cholesterol-7α-hydroxylase expression. FGF, MGF, FPS and MIP decreased the HFHC diet-increased area under the curve (AUC) of serum cholesterol. FGF and FPS further decreased AUC of serum triglycerides. When evaluating the redox status of erythrocytes, FPS and MIP increased non-protein sulfhydryl (NP-SH) groups, reduced glutathione (GSH) and catalase activity and FPS further increased GSH peroxidase activity. In the liver, MGF increased NP-SH groups and GSH and decreased triglycerides content. FPS, MIP and BEP decreased oxidized GSH and triglycerides content. Moreover, all treatments alleviated HFHC diet-increased LDL oxidation. CONCLUSIONS: Fruiting bodies of G. frondosa may improve hypercholesterolaemia via increased bile acid synthesis. Additionally, fermented biomass and polysaccharides of G. frondosa may have the potential to prevent hepatic lipid accumulation.

Gluten sensitivity: associated sporadic cerebellar ataxia in Taiwan.

PURPOSE: Gluten sensitivity (GS) is related to the pathogenesis of sporadic or hereditary ataxia. METHODS: Total of 194 healthy controls and patients with either hereditary ataxia (n=207) or sporadic ataxia (n=361) were tested for the circulating gluten-related autoantibodies which serve as biomarkers to interpret the existence of GS. RESULTS: The incidences of GS in each population were 1% in normal subjects, 2% in hereditary ataxia patients and 9% in sporadic ataxia patients. High serum level of anti-gliadin IgG/IgA and t-transglutaminase IgA were disclosed at the sporadic ataxia patients compared with normal subjects. However, the anti-gliadin IgG is more specific to the disease of sporadic ataxia. CONCLUSION: Relatively higher incidence of GS was found in the population of sporadic ataxia patients but not in either normal subjects or hereditary ataxia patients in Taiwan. Anti-gliadin IgG still is a very powerful indicator to implicate the immune-related sporadic ataxia and we conclude that GS-related sporadic ataxia exists in Taiwan with linkage to autoimmune events.

Prevalence of Blomia tropicalis in wheezing children in central Taiwan.

BACKGROUND AND PURPOSE: In a previous study, we found that wheezing children in rural central Taiwan had a significantly lower average sensitization rate to Dermatophagoides pteronyssinus (Der p) than those in Taipei city. We propose that Blomia tropicalis (Blo t) might be the major mite allergen in rural central Taiwan. METHODS: Using the preserved sera from our previous study, we retrospectively measured specific immunoglobulin E (IgE) antibody to Blo t and analyzed the correlation between Blo t and Der p in wheezing children in rural central Taiwan. A total of 2206 children with physician-diagnosed asthma and wheezing were enrolled and categorized among five age groups. The sensitization rate and level of specific IgE antibody to Blo t were analyzed. RESULTS: The age-specific sensitization rates and the level of specific IgE antibody to either Blo t or Der p increased progressively with increasing age, being greatest in the age group 8 to 12 years. A significant positive correlation existed between sensitization rate and age for both Blo t and Der p (p=0.001). Specific IgE antibody to Blo t was undetectable in patients younger than 1.5 years. A significant positive correlation also existed between age and anti-Blo t IgE antibody level (p<0.003). However, the allergen-specific IgE level was lower for Blo t than Der p (p<0.005) in all age groups. CONCLUSIONS: Blo t might be the major mite allergen to associated with early wheeze and atopic asthma in rural central Taiwan.

Host defense against Salmonella and rotaviral gastroenteritis: a serial study of transcriptional factors and cytokines.

BACKGROUND AND PURPOSE: Common etiologies of acute enterocolitis in childhood include the intracellular pathogens Salmonella and rotavirus, along with extracellular pathogens. In order to elucidate differentiating immunologic parameters in patients with acute gastroenteritis of different etiologies, we investigated interferon (IFN)-gamma, interleukin (IL)-12, and T-bet of T-helper type 1 subsets, and IL-4 and GATA-3 of T-helper type 2 subsets. METHODS: From June 1, 2003 to December 31, 2003, 32 patients with acute gastroenteritis were enrolled. Sequential heparinized blood samples were obtained on the day of presentation (day 1) and on day 3 of hospitalization. Using reverse transcriptase-polymerase chain reaction, the mean ratios of IFN-gamma, T-bet and IL-12 mRNA levels relative to beta-actin were determined. RESULTS: Salmonella infections induced stronger IFN-gamma and T-bet responses than either rotavirus infection or other enterocolitis (p<0.05). However, poor IL-12 response in Salmonella infections implied failed T-helper type 1 immunity, and probably accounted for the prolonged clinical course. In contrast, by day 3 of hospitalization, most patients with rotavirus enterocolitis were symptom-free. CONCLUSIONS: IL-12 is the key factor in determining host response against and, hence, disease activity of Salmonella infections.

Massive expansion of EBV+ monoclonal T cells with CD5 down regulation in EBV-associated haemophagocytic lymphohistiocytosis.

Haemophagocytic lymphohistiocytosis (HLH) comprises primary and secondary forms; the secondary form is most commonly triggered by the Epstein-Barr virus (EBV; EBV-HLH). Patients with EBV-HLH usually exhibit oligoclonal or monoclonal T cell proliferation, which may mimic T cell lymphoproliferative disorder (T-LPD). This article reports on EBV-HLH in a 17-month-old girl with an extreme surge of reactive T lymphocytosis (absolute count 167x10(9)/l) with CD5 down regulation. Bone marrow aspirate and trephine contained florid haemophagocytosis and massive infiltration of CD3+ Epstein-Barr virus-encoded RNA+ lymphocytes, as seen by double labelling. These lymphocytes were monoclonal for EBV and T cell receptor gamma chain gene rearrangement. The patient responded dramatically to intravenous immunoglobulin, interferon alpha2b, ganciclovir and prednisolone, suggesting restoration of her immune system and eradication of the clonal T cells through these immunoregulatory agents. Thus, careful clinicopathological correlation is warranted in the interpretation of immunophenotyping and clonality data in T cell proliferation in association with EBV-HLH to avoid erroneous diagnosis of T-LPD.

Hyperglycemia and advanced glycation end products (AGEs) suppress the differentiation of 3T3-L1 preadipocytes.

Aging is characterized by mild hyperglycemia and accumulation of advanced glycation end products (AGEs). Effects of chronic exposure to hyperglycemia or AGEs on the adipogenic differentiation of 3T3-L1 preadipocytes remain unclear. We examined the chronic effect of AGEs and high glucose on the differentiation of 3T3-L1 cells by culturing 3T3-L1 cells in the presence of AGEs or 25 mM glucose for 1 month. Chronic incubation of 3T3-L1 cells with AGEs or high glucose blocked their differentiation into mature adipocytes as evidenced by reduced levels of adipocyte markers such as accumulated oil droplets, GPDH, aP2, adiponectin and of adipogenesis regulators PPARγ and C/EBPα. Levels or activities of Src, PDK1, Akt, and NF-κB were higher in AGEs- and high glucose-treated cells than those in 3T3-L1 cells. Levels of Bcl-2 were elevated in AGEs- and high glucose-treated cells, and were attenuated by inhibitors of PI3-kinase, Akt and NF-κB. Moreover, adipogenesis was attenuated in 3T3-L1 cells stably expressing Bcl-2 or YAP. These results suggest that chronic AGEs and high glucose treatments up-regulate Bcl-2 and YAP via the Akt-NF-κB pathway and impair adipogenesis.

Caveolin-1 Secreted from Adipose Tissues and Adipocytes Functions as an Adipogenesis Enhancer.

OBJECTIVE: Caveolin-1 (Cav-1) is expressed abundantly in adipose tissue and involved in many physiological processes. While Cav-1 has been reported to be secreted in pancreatic acinar cells and LNCaP prostate cancer cells, its secretion from adipose tissue awaits investigation. METHODS: Cav-1 secretion from 3T3-L1 adipocytes and fat tissues from normal chow diet- and high-fat diet (HFD)-fed mice was measured. Functions and uptake of secreted Cav-1 proteins were assessed by adding Cav-1 back to preadipocytes and LNCaP cells. RESULTS: Cav-1 secretion was evident in adipose tissues and were substantially promoted in HFD-fed mice. Cav-1 was detectable in the conditioned media of 3T3-L1 adipocytes but not preadipocytes. Hypertrophied adipocytes induced by glucose and fatty acids secreted more Cav-1, suggesting that hypertrophied adipocytes were responsible for enhanced Cav-1 secretion in obese mice. Secreted Cav-1 was taken up by preadipocytes and LNCaP cells. 3T3-L1 preadipocytes overexpressing Cav-1 were better differentiated, suggesting that secreted Cav-1 may promote adipogenesis. Hypertrophied 3T3-L1 adipocytes enhanced ERK1/2 activation, and the attenuation of ERK1/2 activity by PD98059 inhibited Cav-1 secretion. CONCLUSIONS: Cav-1 is actively secreted from adipocytes as a putative adipogenesis enhancer. Hypertrophied adipocytes secrete Cav-1 via ERK1/2-dependent mechanisms to promote adipogenesis, thus establishing a vicious cycle.

Higher incidence of Dermatophagoides pteronyssinus allergy in children of Taipei city than in children of rural areas.

BACKGROUND AND PURPOSE: House dust mites are the most common cause of sensitization in wheezing children in most parts of the world. The aim of this study was to investigate prevalence of sensitization and the levels of specific immunoglobulin E (IgE) to Dermatophagoides pteronyssinus (Der p) in wheezing children in Taipei city and central rural Taiwan. METHODS: A total of 3546 children were enrolled in this study. Children were grouped into those living in Taipei city (n = 1340) and those residing in the rural part of central Taiwan (2206). The prevalence of sensitization and level of specific IgE antibody to Der p and cockroach were analyzed by age and geographic area. RESULTS: The results showed significantly higher sensitization rates and mean specific IgE levels to Der p with increasing age of patients in both Taipei city and rural central Taiwan. In addition, children from Taipei city had a significantly higher average sensitization rate of Der p than rural children (p<0.05). The proportion of children sensitized to cockroach increased with age both in Taipei city and central rural Taiwan, but the specific IgE levels to cockroach were not statistically different between the 2 groups (p=0.061). CONCLUSIONS: Sensitivity to aeroallergens varies with age and with geographical location (city vs rural) in asthmatic children. Circulating IgE antibodies against Der p were common at all ages. Cockroach allergen is also associated with recurrent wheezing in Taiwan. Avoidance of indoor aeroallergens such as Der p and cockroach allergen should be an important component in plans for the management of recurrent wheezing.

Somatic mitochondrial DNA mutations in oral cancer of betel quid chewers.

Somatic mitochondrial DNA alteration is a general phenomenon that occurs in cancerous cells. Although numerous mtDNA mutations have been identified in various tumors, the pathogenic significance of these mutations remains unclear. In order to better understand the role of mtDNA mutations in the neoplastic process of oral cancer, the occurrence of mtDNA mutations in oral squamous cell carcinomas was screened by temporal temperature gradient gel electrophoresis (TTGE). The entire mitochondrial genome was amplified with 32 pairs of overlapping primers. The DNA fragments showing different banding patterns between normal and tumor mtDNA were sequenced for the identification of the mutations. Fourteen of 18 (77.8%) tumors had somatic mtDNA mutations with a total of 26 mutations. Among them, 6 were in mRNA coding region. Three were missense mutations (C14F, H186R, T173P) in NADH dehydrogenase subunit 2 (ND2). One frameshift mutation, 9485delC, was in cytochrome c oxidase subunit III. Eight (44%) tumors had insertion or deletion mutations in the np303-309 poly C region of the D-loop. Our results demonstrate that somatic mtDNA mutations occur in oral cancer. The missense and frameshift mutations in the evolutionary conserved regions of the mitochondrial genome may have functional significance in the pathogenesis of oral cancer.

Unilateral agenesis of the internal carotid artery in CHARGE syndrome.

CHARGE syndrome is a multisystemic disorder comprising colobomas, heart defects, choanal atresia, retarded growth and development, genital hypoplasia, ear anomalies and deafness. The CHD7 gene on chromosome 8q12.1 was recently shown to be a major gene involved in the etiology of this syndrome. We describe a girl with CHARGE syndrome who had a novel mutation of CHD7 associated with agenesis of the left internal carotid artery. She had presented with recurrent episodes of photophobia and vomiting since the age of 6 years. Since her symptoms were well controlled by cyproheptadine, migraine-like attacks were considered. CHD7 molecular confirmation in this patient provides further evidence to support the occurrence of a vascular anomaly suggested from animal models of CHARGE syndrome with molecular delineation. We report this case to emphasize the importance of neurologic signs of photophobia and to highlight the broad clinical variability in this pleiotropic disorder.

Daily urinary interleukin-11 excretion correlated with proteinuria in IgA nephropathy and lupus nephritis.

Interleukin-11 (IL-11) is a multifunctional cytokine with both thrombopoietic and anti-inflammatory effects. In an animal study IL-11 was shown to reduce proteinuria in mice with necrotizing glomerulonephritis. The purpose of this current study is to explore the role of IL-11 in human glomerulonephritis. Subjects of this study were patients with proteinuria (daily urine protein excretion >40 mg/m2 per hour) and underlying pathology of IgA nephropathy (IgAN) (n=20), lupus nephritis (LN) (n=40), and idiopathic nephrotic syndrome (INS) (n=68). Daily urinary IL-11 level was measured by enzyme-linked immunosorbent assay (ELISA). Correlation between urinary IL-11 and urinary protein was determined by Pearson's correlation coefficient. Another five patients with serial data of urinary protein, IL-11 and IL-11 messenger RNA (mRNA) expression in urine sediment are presented. The correlation between urinary IL-11 and daily urinary protein was significant for patients with IgAN (r=0.596, P=0.006) and LN (r=0.630, P<0.001), but not for patients with INS (r=0.030, P=0.812). Serial data revealed the same correlation. Furthermore, the peak of urinary IL-11 mRNA preceded that of urinary IL-11. We conclude that daily urinary IL-11 excretion is correlated with urinary protein loss in nephritis having local T helper (Th)1 predominant immune response, such as IgAN and LN. Local IL-11 production may serve as a counter cytokine against Th1-mediated inflammation.

Induced apoptosis of TH2 lymphocytes in asthmatic children treated with Dermatophagoides pteronyssinus immunotherapy.

Allergen-specific immunotherapy (IT) has been effectively used for the treatment of asthma. Allergen specific IT induced immune tolerance with induction of TH2 cells anergy remain to be clarified. The aim of this study was to evaluate whether the mite allergen Dermatophagoides pteronyssinus (Dpt) specific IT serially decreased IL-4+/CD4+ (TH2) lymphocytes and induced apoptosis of TH2 lymphocytes in asthmatic children. Sixty Dpt-sensitive asthmatic children were randomly assigned to a received IT and an untreated group. Dermatophagoides pteronyssinus specific IT treated patients were examined at three time points: before IT, after 6 months of an increased dose phase and with maximum tolerated doses after 1 yr. Peripheral blood mononuclear cells (PBMC) were isolated and cultured for 48 h for cellular staining with CD4+, CD45RO cell phenotypes and interleukin (IL)-4 and interferon-gamma expression by fluorescence monoclonal antibodies. Apoptosis was measured using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) method. A simultaneous flow cytometric study using the same permeabilized cell was examined to determine whether apoptosis occurred preferentially in TH2 lymphocytes. The data demonstrated that Dpt specific IT decreased Dpt-specific IgE levels (p < 0.01) after 1 yr of treatment. In addition, decreased CD4+IL-4+ TH2 cells with increased CD4+IFN-gamma+ TH(1) cells were observed at 6 months and 1 yr after IT treatment (p < 0.05). At the same time, apoptosis of CD4+IL-4+ TH2 lymphocytes in the IT group had increased after 1 yr of treatment when compared with the results before treatment (p < 0.001) and after 6 months of treatment (p = 0.046). In addition, CD45RO cells apoptosis mainly occurred after 6 months of IT treatment and after 1-year period of IT treatment (p < 0.05). All of the data suggested that Dpt specific IT decreased Dpt specific IgE and CD4+IL-4+ TH2 lymphocytes with induction apoptosis of CD4+IL-4+ TH2 lymphocytes subsets serially.

Immunoglobulin G profiles in different forms of periodontitis.

BACKGROUND: Races and Gm(23) allotypes can modify the serum levels of IgG subclasses. The average serum levels of IgG subclasses of African-Americans have been reported to be higher than those of Caucasians in both healthy young adults and patients with aggressive periodontitis; Gm(23)-positive subjects generally had higher IgG2 levels than Gm(23)-negative subjects. OBJECTIVE: We examined serum immunoglobulin G (IgG) concentrations in Taiwanese patients with different forms of periodontitis. METHODS: The serum levels of four IgG subclasses were determined by enzyme-linked immunosorbent assay and Gm(23) allotypes were verified by radial immunodiffusion tests in 50 patients with chronic periodontitis, 30 patients with aggressive periodontitis, and 74 healthy controls. RESULTS: There were no differences in the concentrations of four IgG subclasses in patients with chronic periodontitis compared with age-matched controls. However, in subjects younger than 35 years, levels of IgG2 were significantly elevated in patients with aggressive periodontitis compared with controls. We also found significant differences in IgG2 levels within the control group when stratified by age (< or = 35 years and > 35 years). Gm(23) allotypes were not correlated with the serum levels of IgG2 in either patient group. CONCLUSION: Microbial challenge might not provoke significant changes in systemic IgG response in patients with chronic periodontitis. However, in patients with aggressive periodontitis, IgG2 levels were increased when compared with age-matched controls. Gm(23) allotypes had no influence on IgG2 levels in well-established generalized chronic periodontitis or aggressive periodontitis.

Gm (23) allotypes and Fcgamma receptor genotypes as risk factors for various forms of periodontitis.

OBJECTIVES: Given the diversity of the distribution of the Gm (23) allotypes and FcgammaR genotypes in different ethnic groups, it was our purpose to examine their clinical significance in periodontitis in Taiwan. MATERIAL & METHODS: Genomic DNA of 50 patients with chronic periodontitis (CP), 30 patients with generalized aggressive periodontitis (G-AP) and 74 healthy controls were harvested. The Gm (23) allotypes were determined by radial immunodiffusion test, and the FcgammaR IIa (CD32) and IlIb (CD16) genotypes were determined by polymerase chain reaction-based allele-specific oligonucleotide hybridization. RESULTS: The overall carrier rate of the Gm (23+) allotype was higher than 85%, and the Gm (23-) allotype was statistically over-represented in patients with CP compared to the controls. There were no differences in the distributions of the three genotypes of FcgammaR IIa and IIIb among the three tested groups. The frequency of the R131 allele of the FcgammaR IIa polymorphisms was higher in G-AP than in CP when R/H allelic frequencies (p = 0.01) were examined by the chi2 test. CONCLUSION: The Gm (23-) allotype might be a potential risk factor for CP. Although the R131 allele of FcgammaR IIa occurred more frequently in G-AP than in CP, its clinical significance could not be justified in this study.

Novel heteroplasmic frameshift and missense somatic mitochondrial DNA mutations in oral cancer of betel quid chewers.

Mitochondrial DNA (mtDNA) has been proposed to be involved in carcinogenesis because of its high susceptibility to oxidative DNA damage and limited repair mechanisms. For investigation of the potential role of somatic mtDNA mutations in the tumorigenesis of oral cancer, we screened the occurrence of mtDNA mutations by the temporal temperature gradient gel electrophoresis method. We amplified the entire mitochondrial genome by use of 32 pairs of overlapping primers, and to identify the mutations, we sequenced DNA fragments showing different banding patterns between normal and tumor mtDNA. Fourteen of eighteen (77.8%) oral carcinomas displayed somatic mtDNA mutations, with a total of 26 mutations. Among them, six were in the mRNA coding region. Three were missense mutations (C14F, H186R, T173P) in NADH dehydrogenase subunit 2, and one was a frameshift mutation, 9485delC, in cytochrome c oxidase subunit III. Eight (44%) tumors had insertion or deletion mutations in the nucleotide position 303-309 poly C region of the D-loop. Multiple large deletions were also observed. Our results demonstrate that somatic mtDNA mutations occur in oral cancer. Some missense and frameshift mutations may play an important role in the tumorigenesis of this carcinoma. More extensive biochemical and molecular studies will be necessary for determining the pathologic effect of these somatic mutations.