ERK-dependent IL-6 autocrine signaling mediates adaptive resistance to pan-PI3K inhibitor BKM120 in head and neck squamous cell carcinoma.

Hyperactivation of phosphatidylinositol 3-kinase (PI3K) pathway occurs frequently in head and neck squamous cell carcinoma (HNSCC). However, clinical outcomes of targeting the PI3K pathway have been underwhelming. In present study, we investigated the resistant mechanisms and potential combination therapeutic strategy to overcome adaptive resistance to PI3K inhibitor in HNSCC. Treatment of NVP-BKM120, a pan-PI3K inhibitor, led to upregulation of interleukin-6 (IL-6) and subsequent activation of either extracellular signal-regulated kinase (ERK) or signal transducers and activators of transcription 3 (STAT3), causing modest antitumor effects on the growth of HNSCC cells. Blockade of autocrine IL-6 signaling with siRNA or neutralizing antibody for IL-6 receptor (IL-6R) completely abolished NVP-BKM120-induced activation of ERK and STAT3 as well as expression of c-Myc oncogene, which resulted in enhanced sensitivity to NVP-BKM120. Moreover, when compared with a pharmacologic inhibitor or silencing of STAT3, trametinib, a MEK inhibitor, in combination with NVP-BKM120 yielded more potent anti-proliferative effects by inhibiting S phase transition, arresting cells at G0/G1 phase, and downregulating IL-6 and c-Myc expression. Furthermore, as compared with either agent alone, combination of NVP-BKM120 with trametinib or tocilizumab, a humanized anti-IL-6R antibody, significantly suppressed tumor growth in NVP-BKM120-resistant patient-derived tumor xenograft (PDTX) models, which was also confirmed in PDTX-derived cell lines. Collectively, these results suggested that IL-6/ERK signaling is closely involved in adaptive resistance of NVP-BKM120 in HNSCC cells, providing a rationale for a novel combination therapy to overcome resistance to PI3K inhibitors.

Common developmental genome deprogramming in schizophrenia - Role of Integrative Nuclear FGFR1 Signaling (INFS).

The watershed-hypothesis of schizophrenia asserts that over 200 different mutations dysregulate distinct pathways that converge on an unspecified common mechanism(s) that controls disease ontogeny. Consistent with this hypothesis, our RNA-sequencing of neuron committed cells (NCCs) differentiated from established iPSCs of 4 schizophrenia patients and 4 control subjects uncovered a dysregulated transcriptome of 1349 mRNAs common to all patients. Data reveals a global dysregulation of developmental genome, deconstruction of coordinated mRNA networks, and the formation of aberrant, new coordinated mRNA networks indicating a concerted action of the responsible factor(s). Sequencing of miRNA transcriptomes demonstrated an overexpression of 16 miRNAs and deconstruction of interactive miRNA-mRNA networks in schizophrenia NCCs. ChiPseq revealed that the nuclear (n) form of FGFR1, a pan-ontogenic regulator, is overexpressed in schizophrenia NCCs and overtargets dysregulated mRNA and miRNA genes. The nFGFR1 targeted 54% of all human gene promoters and 84.4% of schizophrenia dysregulated genes. The upregulated genes reside within major developmental pathways that control neurogenesis and neuron formation, whereas downregulated genes are involved in oligodendrogenesis. Our results indicate (i) an early (preneuronal) genomic etiology of schizophrenia, (ii) dysregulated genes and new coordinated gene networks are common to unrelated cases of schizophrenia, (iii) gene dysregulations are accompanied by increased nFGFR1-genome interactions, and (iv) modeling of increased nFGFR1 by an overexpression of a nFGFR1 lead to up or downregulation of selected genes as observed in schizophrenia NCCs. Together our results designate nFGFR1 signaling as a potential common dysregulated mechanism in investigated patients and potential therapeutic target in schizophrenia.

Comparative linkage maps suggest that fission, not polyploidy, underlies near-doubling of chromosome number within monkeyflowers (Mimulus; Phrymaceae).

Changes in chromosome number and structure are important contributors to adaptation, speciation and macroevolution. In flowering plants, polyploidy and subsequent reductions in chromosome number by fusion are major sources of chromosomal evolution, but chromosome number increase by fission has been relatively unexplored. Here, we use comparative linkage mapping with gene-based markers to reconstruct chromosomal synteny within the model flowering plant genus Mimulus (monkeyflowers). Two sections of the genus with haploid numbers ≥ 14 have been inferred to be relatively recent polyploids because they are phylogenetically nested within numerous taxa with low base numbers (n=8-10). We combined multiple data sets to build integrated genetic maps of the M. guttatus species complex (section Simiolus, n=14) and the M. lewisii group (section Erythranthe; n=8), and then aligned the two integrated maps using >100 shared markers. We observed strong segmental synteny between M. lewisii and M. guttatus maps, with essentially 1-to-1 correspondence across each of 16 chromosomal blocks. Assuming that the M. lewisii (and widespread) base number of 8 is ancestral, reconstruction of 14 M. guttatus chromosomes requires at least eight fission events (likely shared by Simiolus and sister section Paradanthus (n=16)), plus two fusion events. This apparent burst of fission in the yellow monkeyflower lineages raises new questions about mechanisms and consequences of chromosomal fission in plants. Our comparative maps also provide insight into the origins of a chromosome exhibiting centromere-associated female meiotic drive and create a framework for transferring M. guttatus genome resources across the entire genus.

Effects of light on secondary metabolism and fungal development of Fusarium graminearum.

AIMS: The objectives of this study were (i) to characterize white-collar (WC) orthologues of the filamentous fungus Fusarium graminearum, (ii) to investigate light-responsive phenotypes by the deletion of Fgwc-1 and Fgwc-2 genes and (iii) to examine the roles of those genes in constant light and darkness in relation to secondary metabolite synthesis and development. METHODS AND RESULTS: Production of secondary metabolites and asexual/sexual development of deletion mutants, ΔFgwc-1 and ΔFgwc-2, were assessed in constant light and darkness compared to the wild-type strain. The results showed that deletion of Fgwc-1 and Fgwc-2 impaired early onset of carotenogenesis, photoreactivation and the maturity of perithecia during sexual development. Conidiation of the ΔFgwc-1 and ΔFgwc-2 mutants was derepressed in constant light, but not in darkness. Moreover, the individual mutants produced more aurofusarin and trichothecenes than the wild-type strain in both constant light and darkness. CONCLUSIONS: Both Fgwc-1 and Fgwc-2 are required for light-dependent processes in F. graminearum, whereas light-independent processes such as aurofusarin and trichothecene biosynthesis are derepressed by deletion of these genes. Thus, Fgwc-1 and Fgwc-2 play roles as positive and negative regulators, depending on the requirement of light for biological activity. SIGNIFICANCE AND IMPACT OF THE STUDY: These results will extend the knowledge of the photobiology of Fusarium graminearum and will increase current understanding of light regulatory mechanisms mediated by white collar in secondary metabolism and fungal development.

First Report of Neocosmospora vasinfecta var. vasinfecta Causing Soybean Stem Rot in South Korea.

In September 2010, stem rot symptoms were observed on soybean plants (cv. Daepungkong) growing in a field located at Daegu (35.52° N, 128.35° E), South Korea. The first noticeable symptoms, observed on the top leaves, were difficult to distinguish from those of sudden death syndrome (SDS). However, after splitting the stems of symptomatic plants, typical stem rot symptoms appeared as reddish-brown to dark-brown discoloration of the pith. Stem lesions extended 15 to 20 cm upward from the soil surface. To isolate the causal agent, sections of diseased stems were surface disinfected with 1% sodium hypochlorite, placed on potato dextrose agar (PDA) containing streptomycin sulfate, and incubated at 25°C with a 12-h light regime. Two isolates were obtained (SSLNV17 and SSLNV18). Mycelia were white and floccose. Conidia (4.5 to 11.2 × 2.2 to 3.4 µm) were cylindrical to oblong-ellipsoidal, hyaline, and one-celled. Both isolates produced abundant perithecia after 3 to 4 weeks. Perithecia (205 to 331 mm in diameter) were orange to red, globose and ostiolate, with a short neck (80 to 126 mm in diameter). Unitunicate asci (88.6 to 115.3 × 14.5 to 17.3 mm) were cylindrical to clavate, with a short stalk (6.0 to 9.5 × 5.0 to 6.8 mm), and eight spores. Ascospores (13.3 to 17.5 × 10.7 to 12.7 mm) were uniseriately arranged, globose to oval, one-celled, and hyaline to pale brown, with walls with a rugose ornamentation. These morphological features are consistent with those of Neocosmospora vasinfecta var. vasinfecta (1). The internal transcribed spacer (ITS) region, partial translation elongation factor 1-alpha (EF1-α), and ß-tubulin genes of rDNA of the two isolates were sequenced using primers ITS4/ITS5 (GenBank Accession Nos. KF662732 and KF662733), EF1-728F/EF1-986R (KF758839 and KF758840), and Bt2a/Bt2b (KF771004 and KF771005), respectively. Sequences of the ITS region, EF1-α, and ß-tubulin genes of both isolates showed 99% similarity with several reported N. vasinfecta strains by BLAST analysis. Both morphological and sequence analyses confirmed that the two isolates were N. vasinfecta var. vasinfecta. Pathogenicity tests of both isolates were performed on 15 three-week-old seedlings of soybean cv. Williams inoculated with a spore suspension containing 1.0 × 106 spores/ml, using stem puncture inoculation procedure under controlled conditions (4). Control plants were inoculated in the same way with sterile water. The results were observed by splitting the stem longitudinally and checking for discoloration of the pith 4 to 5 weeks after inoculation. Reddish-brown to dark-brown discoloration was observed in the stem pith of inoculated plants, with occasional chlorosis of the leaves. Moreover, numerous orange-red perithecia were produced on the inoculated stems. However, no symptoms were visible on control plants. The pathogen was re-isolated from the diseased plants, confirming Koch's postulates. Neocosmospora stem rot of soybean was first discovered in Japan and since then it has been reported in the United States and China (2,3,4). To our knowledge, this is the first record of soybean stem rot caused by N. vasinfecta var. vasinfecta in Korea. Our report indicates that Neocosmospora stem rot is a new threat to soybean production in Korea. References: (1) P. F. Cannon and D. L. Hawksworth. Trans. Br. Mycol. Soc. 82:673, 1984. (2) Y. Gai et al. Plant Dis. 95:1031, 2011. (3) F. A. Gray et al. Plant Dis. 64:321, 1980. (4) D. V. Phillips. Phytopathology 62:612, 1972.

Novel Mutation of IL2RG Gene in a Korean Boy With X-linked Severe Combined Immunodeficiency

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Intention to comply with post-exposure management among nurses exposed to blood and body fluids in Taiwan: application of the theory of planned behaviour.

Nurses are at significant risk from occupationally acquired bloodborne virus infections following a needlestick and sharps injury. This study aimed to apply the theory of planned behaviour (TPB) to predict nurses' intention to comply with occupational post-exposure management. A cross-sectional survey was applied to select registered nurses who worked in human immunodeficiency virus (HIV)-designated hospitals. An anonymous, self-administered questionnaire based on the TPB was distributed to 1630 nurses and 1134 (69.5%) questionnaires were returned. From these, a total of 802 nurses (71%) reported blood and body fluid exposure incidents during 2003-2005 and this group was used for analysis. Only 44.6% of the 121 exposed nurses who were prescribed post-exposure prophylaxis (PEP) by infectious disease doctors returned to the clinic for interim monitoring, and only 56.6% of exposed nurses confirmed their final serology status. Structural equation modelling was used to test the TPB indicating perceived behavioural control (the perception of the difficulty or ease of PEP management, ß=0.58), subjective norm (the perception of social pressure to adhere to PEP, ß=0.15), and attitudes (ß=0.12) were significant direct effects on nurses' intention to comply with post-exposure management. The hypothesised model test indicated that the model fitted with the expected relationships and directions of theoretical constructs [χ(2) (14, N=802)=23.14, P=0.057, GFI=0.987, RMSEA=0.039]. The TPB model constructs accounted for 54% of the variance in nurses' intention to comply with post-exposure management. The TPB is an appropriate model for predicting nurses' intention to comply with post-exposure management. Healthcare facilities should have policies to decrease the inconvenience of follow-up to encourage nurses to comply with post-exposure management.

Tendon adhesion measured by a video-assisted gliding test in a chicken model.

We developed a video-assisted gliding test to evaluate the gliding force and the flexion angle with unrestricted joint motion. Tendon adhesion was induced in a chicken model of flexor digitorum profundus (FDP) injury at the annular pulley region of the long toe. The chicken feet were harvested immediately after injury, and 2 weeks and 6 weeks after injury. During the gliding test, the injured FDP was pulled for 15 mm then returned to its initial position. The test was recorded using a video camera and registered to the gliding test mechanical data. The maximum flexion angle and gliding resistance were calculated. The maximum flexion angle was significantly decreased from 78 (SD 10) in controls to 42 (SD 22) in tendons with injury, while gliding resistance was significantly increased in week 2 (0.06, SD 0.05) and week 6 (0.07, SD 0.01) after injury.

Lack of O-polysaccharide enhances biofilm formation by Bradyrhizobium japonicum.

AIMS: To reveal the effects of the O-polysaccharide antigen of Bradyrhizobium japonicum LPS on biofilm formation and motility. METHODS AND RESULTS: Wild type and O-antigen-deficient mutant strains of B. japonicum were tested for biofilm formation on polyvinyl chloride (PVC) surfaces and motility on semi-solid (0.3%) agar media. After 7 days of incubation, the amount of biofilms formed by the mutant was c. 3.5-fold greater than that of the wild type. Unlike biofilm formation, the motility assay revealed that the mutant strain was less motile than the wild type. CONCLUSIONS: This study shows enhanced biofilm formation and decreased motility by the O-antigen-deficient mutant, suggesting that the lack of the O-polysaccharide of the rhizobial LPS is associated with biofilm-forming ability and movement. SIGNIFICANCE AND IMPACT OF THE STUDY: LPS plays an important role in both pathogenic and beneficial bacteria. It has also been reported that LPS deficiency negatively affects biofilm formation. However, our results demonstrate that the O-antigen-deficient mutant enhances biofilm formation, presumably through a significant increase in hydrophobicity. It is notable that the hydrophobicity of cell walls might be a key regulator in controlling biofilm development in B. japonicum.

Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells.

BACKGROUND: Stathmin1 is a microtubule-regulating protein that has an important role in the assembly and disassembly of the mitotic spindle. The roles of stathmin1 in carcinogenesis of various cancers, including prostate and breast cancer, have been explored. However, its expression and roles in gastric cancer have not yet been described. METHODS: Stathmin1 expression in paraffin-embedded tissue sections from 226 patients was analysed by immunohistochemistry. Roles of stathmin1 were studied using a specific small interfering RNA (siRNA). RESULTS: The expression of stathmin1 was positively correlated with lymph node metastasis, TNM stages and vascular invasion, and negatively with recurrence-free survival, in the diffuse type of gastric cancer. The median recurrence-free survival in patients with a negative and positive expression of stathmin1 was 17.0 and 7.0 months, respectively (P=0.009). When the expression of stathmin1 was knocked down using siRNA, the proliferation, migration and invasion of poorly differentiated gastric cancer cells in vitro were significantly inhibited. Moreover, stathmin1 siRNA transfection significantly slowed the growth of xenografts in nude mice. CONCLUSION: These results suggest that stathmin1 can be a good prognostic factor for recurrence-free survival rate and is a therapeutic target in diffuse-type gastric cancer.

UV/TiO2 and UV/TiO2/chemical oxidant processes for the removal of humic acid, Cr and Cu in aqueous TiO2 suspensions.

The objective of this study was to investigate the treatment efficiency of UV/TiO2 and UV/TiO2/chemical oxidant processes for the removal of humic acid and hazardous heavy metals in aqueous TiO2 suspensions. The reaction rate (k) of humic acid and hazardous heavy metals by UV/TiO2 was higher than that of UV illumination alone or TiO2 alone. The removal efficiency for humic acid and Cr(VI) at acid or neutral pH values was higher than that at basic pH values. However, the removal efficiency for Cu(II) at acid pH values was smaller compared with that at neutral or basic pH values. The reaction rate (k) of humic acid and hazardous heavy metals in the TiO2 concentration range of 0.1-0.3 g l(-1) increased with increasing TiO2 dosage. However, amounts higher than a TiO2 dosage of 0.3 g l(-1) reduced the removal efficiency for humic acid and hazardous heavy metals because of the shielding effect on the UV light penetration in the aqueous solution caused by the presence of excessive amounts of TiO2. The addition of oxidants to the UV/TiO2 system showed an increase in degradation efficiency for the treatment of humic acid and hazardous heavy metals. The optimal concentration of oxidants was: H2O2 50 mg l(-1), O3 20 g m(-3) and K2S2O8 50 mg l(-1), respectively. The degradation efficiency of UV/TiO2/oxidant systems for the removal of humic acid and hazardous heavy metals was much greater when H2O2 was used as the oxidant.

Complete nucleotide sequence of Dendrocalamus latiflorus and Bambusa oldhamii chloroplast genomes.

Although bamboo is one of the most important woody crops in Asia, information on its genome is still very limited. To investigate the relationship among Poaceae members and to understand the mechanism of albino mutant generation in vitro, the complete chloroplast genome of two economically important bamboo species, Dendrocalamus latiflorus Munro and Bambusa oldhamii Munro, was determined employing a strategy that involved polymerase chain reaction (PCR) amplification using 443 novel primers designed to amplify the chloroplast genome of these two species. The lengths of the B. oldhamii and D. latiflorus chloroplast genomes are 139,350 and 139,365 bp, respectively. The organization structure and the gene order of these two bamboos are identical to other members of Poaceae. Highly conserved chloroplast genomes of Poaceae facilitated sequencing by the PCR method. Phylogenetic analysis using both chloroplast genomes confirmed the results obtained from studies on chromosome number and reproductive organ morphology. There are 23 gaps, insertions/deletions > 100 bp, in the chloroplast genomes of 10 genera of Poaceae compared in this study. The phylogenetic distribution of these gaps corresponds to their taxonomic placement. The sequences of these two chloroplast genomes provide useful information for studying bamboo evolution, ecology and biotechnology.

Characterization of the major allergens of Pachycondyla chinensis in ant sting anaphylaxis patients.

BACKGROUND: The ant species Pachycondyla chinensis, which has spread from Far Eastern Asia to New Zealand and North America, induces anaphylactic reactions in human with its sting. However, the major allergens of P. chinensis have not yet been characterized. METHODS: We selected seven patients with histories of anaphylaxis induced by P. chinensis. Two-dimensional electrophoresis (2-DE) was used to identify the major allergens. We subsequently performed Western blots for P. chinensis-specific IgEs, N-terminal amino acid sequencing, ESI-MS/MS, and RT-PCR using primers based on the N-terminal sequence. RESULTS: Six of the anaphylactic subjects had an IgE specific to a 23 kDa allergen of P. chinensis. Two candidates for major allergens, 23 kDa (pI 8.7) and 25 kDa (pI 6.2), were revealed by 2-DE using P. chinensis-specific IgE immunoblotting. In N-terminal sequencing and ESI-MS/MS analysis, 23 kDa (pI 8.7) and 25 kDa (pI 6.2) allergens, belonging to the protein families of antigen 5, were identified and share marked amino acid sequence similarity. The 23 kDa allergen is 206 amino acids in length and homology searches showed 54.0% and 50.0% homology with Sol i 3 and Ves v 5, respectively. CONCLUSION: The major allergens of P. chinensis are 23 kDa (pI 8.7) and 25 kDa (pI 6.2) proteins that belong to the antigen 5 family of proteins.

Clinical and genetic analysis of a Korean patient with late-onset X-linked adrenal hypoplasia congenita and hypogonadotropic hypogonadism: identification of a novel mutation in the NR0B1 gene.

Adrenal hypoplasia congenita (AHC) is caused by mutations in the NR0B1 gene on chromosome Xp21.3-p21.2. It manifests as X-linked primary adrenal failure in early infancy or childhood and as hypogonadotropic hypogonadism (HHG) at puberty. Although studies of AHC patients and mutations in the NR0B1 gene have been reported throughout the world, there has previously been only one other case report from Korea. We encountered a 23-year old Korean male with delayed-onset AHC/HHG who had been previously diagnosed with adrenal insufficiency of unknown aetiology at age 13 years. Delayed puberty and incomplete HHG were observed. Direct sequencing of the NR0B1 gene revealed the patient to have a novel insertion mutation (c.959_960insT; Leu321ProfsX68). Although AHC is believed to be rare, it should be considered in a differential diagnosis of patients showing late-onset primary adrenal insufficiency.

Clinical utility of fine-needle aspiration for diagnosis of head and neck lymphoma.

BACKGROUND: Although fine-needle aspiration (FNA) is currently used for the diagnosis of lymphoma, its diagnostic utility in patients with head and neck (HN) lymphoma is unclear. We therefore assessed the utility of initial clinical and FNA diagnoses for HN lymphoma in a clinician's perspective. METHODS: We conducted a retrospective study of total 109 patients with HN lymphoma underwent both FNA and tissue diagnoses from January 2000 through December 2005. The diagnostic sensitivity of FNA was compared with that of histopathology. FNA diagnosis was based on cytomorphology alone in 69 patients and on immunophenotyping plus morphology in 40. RESULTS: On clinical diagnosis, lymphoma was suspected in 54 patients, nonlymphoma/metastatic malignancy in 31, and benign disease in 24. FNA diagnosed lymphoma in 41 patients; suspicious of lymphoma in 23; atypical lymphoma in 20; benign disease in 19; and was nondiagnostic in 6 patients. Diagnostic accuracy of FNA was not significantly improved by repeat core needle biopsy and immunophenotyping. Delay from FNA to tissue diagnosis was significant in the benign FNA-diagnostic group, with a mean 49 days. CONCLUSIONS: The clinical and FNA diagnoses of HN lymphoma may be incomplete and include the potential pitfall of significant diagnostic delay.

Doxapram and aminophylline on bispectral index under sevoflurane anaesthesia: a comparative study.

BACKGROUND AND OBJECTIVE: To evaluate and compare the effect of two clinically available central nervous system stimulants, namely doxapram and aminophylline on arousal from sevoflurane anaesthesia and bispectral index. METHODS: This randomized, double-blind, placebo-controlled, prospective study was conducted in 90 adult females, ASA I-II, scheduled for elective lower abdominal surgeries at Taipei Medical University Hospital. At 5 min before the completion of surgery, under sevoflurane anaesthesia, patients were divided into three groups to receive doxapram 1 mg kg(-1), aminophylline 2 mg kg(-1) or saline placebo intravenous. Standard vital signs, end-tidal CO(2), end-expiratory sevoflurane concentration, bispectral index and neuromuscular blockade were measured plus clinical parameters of recovery from general anaesthesia. RESULTS: Compared with the control group, patients receiving doxapram or aminophylline showed a similarly faster recovery from sevoflurane anaesthesia correlated with increase in bispectral index. CONCLUSION: Intravenous administration of doxapram 1 mg kg(-1) or aminophylline 2 mg kg(-1) hastened the early recovery from sevoflurane anaesthesia. The arousal effect of aminophylline and doxapram appears to be similar.