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Purpose: To determine if gene signatures differentially expressed in head and neck squamous cell carcinomas (HNSCC) are related to alterations in transcription factors nuclear factor-κB (NF-κB) and TP53 previously associated with decreased cell death, response to therapy, and worse prognosis.Experimental Design: Unique gene signatures expressed by HNSCC lines were identified by cDNA microarray, principal components, and cluster analyses and validated by quantitative reverse transcription-PCR (RT-PCR) and in situ hybridization. Bioinformatic analysis of the promoters and ontogeny of these clustered genes was done. Expression of proteins encoded by genes of a putative NF-κB signature, NF-κB p65, and TP53 were examined in HNSCC tissue specimens by immunostaining. Predicted promoter binding and modulation of expression of candidate NF-κB genes and cell survival were evaluated by p65 chromatin immunoprecipitation (ChIP) and small interfering RNA (siRNA) knockdown.Results: Two groups of HNSCC exhibiting distinct gene signatures were identified: cluster A enriched for histone genes, with a higher prevalence of TP53 promoter binding motifs; and cluster B enriched for injury response genes with NF-κB regulatory motifs. Coexpression of cluster B proteins was observed with strong NF-κB phospho-p65 and weak TP53 staining, and NF-κB phospho-p65 was inversely associated with TP53 (P = 0.02). Promoter binding of the NF-κB signature genes was confirmed by p65 ChIP, and down-modulation of their expression and cell death were induced by p65 siRNA.Conclusion: NF-κB promotes expression of a novel NF-κB–related gene signature and cell survival in HNSCC that weakly express TP53, a subset previously associated with inactivated wild-type TP53, greater resistance to chemoradiotherapy, and worse prognosis.

Supplementary Data from A Novel Nuclear Factor-κB Gene Signature Is Differentially Expressed in Head and Neck Squamous Cell Carcinomas in Association with TP53 Status

Supplementary Data from Deficient TP53 Expression, Function, and Cisplatin Sensitivity Are Restored by Quinacrine in Head and Neck Cancer

Purpose: To determine the nature and potential pharmacologic reversibility of deficient TP53 expression and function in head and neck squamous cell carcinomas (HNSCC) with wild-type TP53, previously associated with decreased sensitivity to cisplatin therapy.Experimental Design:TP53 genotype, mRNA and protein expression, TP53-induced p21 expression, and TP53 DNA–binding and reporter gene function were determined in a panel of nine previously characterized HNSCC cell lines from the University of Michigan squamous cell carcinoma (UM-SCC) series. The genotoxic drug doxorubicin and the anti-inflammatory and antimalarial drug quinacrine, previously identified as inducers of TP53, were used to examine the nature and potential reversibility of deficient TP53 expression and function. The specific role of inducible TP53 on function and cellular proliferation was confirmed using selective TP53 inhibitor pifithrin-α or short hairpin RNA knockdown. The capability of quinacrine to sensitize HNSCC to the cytotoxic effects of cisplatin was assessed.Results: UM-SCC cell lines with wild-type TP53 genotype underexpressed TP53 mRNA and protein when compared with normal human keratinocytes or UM-SCC with mutant TP53. Although doxorubicin failed to induce TP53 expression or functional activity, quinacrine induced TP53 mRNA and protein expression, increased TP53 reporter activity and p21 protein expression, and induced growth inhibition in these wild-type TP53 cell lines. Quinacrine-induced TP53 reporter activity and growth suppression were attenuated by pifithrin-α and TP53 short hairpin RNA knockdown. Furthermore, quinacrine sensitized UM-SCC to cisplatin in vitro.Conclusions: Deficient TP53 mRNA and protein expression underlies decreased function in a subset of HNSCC with wild-type TP53 and can be restored together with cisplatin sensitization by quinacrine.

Supplementary Methods, Figures 1-6 from Attenuated Transforming Growth Factor β Signaling Promotes Nuclear Factor-κB Activation in Head and Neck Cancer

Although constitutively activated nuclear factor-κB (NF-κB), attenuated transforming growth factor β (TGFβ) signaling, and TP53 mutations frequently occur in human cancers, how these pathways interact and together contribute to malignancy remains uncertain. Here, we found an association between overexpression of NF-κB–related genes, reduced expression of TGFβ receptor (TβR) subunits and downstream targets, and TP53 genotype in head and neck squamous cell carcinoma (HNSCC). In response to recombinant TGFβ1, both growth inhibition and TGFβ target gene modulation were attenuated or absent in a panel of human HNSCC lines. However, in HNSCC cells that retained residual TGFβ signaling, TGFβ1 inhibited both constitutive and tumor necrosis factor α–stimulated NF-κB activity. Furthermore, HNSCC lines overexpressing mutant (mt) TP53 and human tumor specimens with positive TP53 nuclear staining exhibited reduced TβRII and knocking down mtTP53 induced TβRII, increasing TGFβ downstream gene expression while inhibiting proinflammatory NF-κB target gene expression. Transfection of ectopic TβRII directly restored TGFβ signaling while inhibiting inhibitor κBα degradation and suppressing serine-536 phosphorylation of NF-κB p65 and NF-κB transcriptional activation, linking these alterations. Finally, experiments with TβRII conditional knockout mice show that abrogation of TGFβ signaling promotes the sustained induction of NF-κB and its proinflammatory target genes during HNSCC tumorigenesis and progression. Together, these findings elucidate a regulatory framework in which attenuated TGFβ signaling promotes NF-κB activation and squamous epithelial malignancy in the setting of altered TP53 status. [Cancer Res 2009;69(8):3415–24]

Livestock wastewater is an important reservoir of antibiotic resistance genes (ARGs), with high environmental risks. We investigated the seasonal variations of distribution and removal of swine wastewater originated high-risk tetracycline resistance genes (TRGs) in horizontal subsurface flow constructed wetlands. The effects of exogenous addition of tetracycline (TC) and copper ion (Cu畜禽养殖废水是抗生素抗性基因(ARGs)的重要贮存库,环境风险不容忽视。本研究考察了夏、冬两季养猪废水中高环境风险四环素抗性基因(TRGs)在水平潜流人工湿地中的分布和去除情况,通过外源添加四环素(TC)和铜离子(Cu

Background Antiphospholipid syndrome (APS) is a non-inflammatory autoimmune disorder induced by antiphospholipid antibodies, which occurs exceedingly rarely in pediatric population and even more rarely reported in HIV positive children. Case summary A case of 11 years old boy had a sudden onset of swelling in his left lower leg along with pain which were worsening gradually. Initially, topical ointment was applied for 1 month which were ineffective in reducing pain and swelling. Instead, the symptoms were aggravated and suddenly spread to the proximal thigh, accompanied by dyskinesia of left lower leg. Both color doppler ultrasonography and vascular CT scan of left lower leg revealed deep venous thrombosis. His serum anti-phospholipid antibodies (aPLs) were tested positive. He was a known case of HIV virological failure with substantial HIV viral load (VL) despite receiving regular antiretroviral therapy (ART). His symptoms improved after giving aggressive antithrombotic and high dose corticosteroid treatments. Conclusion When pediatric patients develop thrombotic disease, APS also needs to be ruled out. The autoantibodies levels should be routinely tested to look for recurrent thrombosis in children with HIV/AIDS.

A novel bacterial strain, TLK-CK17T, was isolated from cow dung compost sample. The strain was Gram-stain-negative, non-spore-forming, short rod-shaped, aerobic, and displayed growth at 15–40°C with an optimum at 35°C, with 0–5.0% (w/v) NaCl (optimum 0.5) and at pH 6.5–8.5 (optimum 7.0–7.5). Cells contained summed iso-C16:0, iso-C15:0, and feature 9 (comprising C17:1 ω9c and / or 10-methyl C16:0), as its major cellular fatty acids (>10.0%) and ubiquinone-8 (Q-8) as the exclusively respiratory quinone. The polar lipid profile of strain TLK-CK17T consisted of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The DNA G+C content was 68.2 mol%. On the basis 16S rRNA gene sequence comparison, strain TLK-CK17T showed the highest sequence similarity (98.9%) with L. penaei GDMCC 1.1817T, followed by L. maris KCTC 42381T (98.3%). Furthermore, the average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) between strain TLK-CK17T and closely related strains L. penaei GDMCC 1.1817T, and L. maris KCTC 42381T were 79.9–85.6% and 23.9–29.6%, respectively. Based on presented results, we propose a novel species for which the name Lysobacter chinensis sp. nov. is suggested, with the type strain TLK-CK17T (=CCTCC AB2021257T= KCTC 92122T).

Compost-based organic fertilizers made from animal manures may contain high levels of antibiotic resistance genes (ARGs). However, the factors affecting the abundance and profile of ARGs in organic fertilizers remain unclear. We conducted a national-wide survey in China to investigate the effect of material type and composting process on ARG abundance in commercial organic fertilizers and quantified the contributions of bacterial composition and mobile genetic elements (MGEs) to the structuring of ARGs, using quantitative PCR and Illumina sequencing of 16S rRNA gene amplicons. The tetracycline, sulfonamide, aminoglycoside and macrolide resistance genes were present at high levels in all organic fertilizers. Seven ARGs that confer resistance to clinically important antibiotics, including three β-lactam resistance genes, three quinolone resistance genes and the colistin resistance gene mcr-1, were detected in 8 - 50% the compost samples, whereas the vancomycin resistance gene vanC was not detected. Raw material type had a significant (p 0.001) effect on the ARG abundance, with composts made from animal feces except some cattle feces generally having higher loads of ARGs than those from non-animal raw materials. Composting process type showed no significant (p 0.05) effect on ARG abundance in the organic fertilizers. MGEs exerted a greater influence on ARG composition than bacterial community, suggesting a strong mobility of ARGs in the organic fertilizers. Our study highlights the need to manage the risk of ARG dissemination from agricultural wastes.

In the framework of upper bound theorem of limit analysis, the progressive collapse of shallow rectangular tunnels with double-layer rock mass has been theoretically analyzed based on the three-dimensional (3D) velocity discontinuity surfaces. According to the virtual work principle, the difference theorem and the variation method, the collapse surface of double-layer rock mass is determined based on the Hoek-Brown failure criterion. The formula can be degenerated to a single-layer rock collapsing problem when the rock mass is homogeneous. To estimate the validity of the result, the numerical simulation software PLAXIS 3D is used to simulate the collapse of shallow tunnels with double-layer rock mass, and the comparative analysis shows that numerical results are in good agreement with upper-bound solutions. According to the results of parametric analysis, the potential range of collapse of a double-layer rock mass above a shallow cavity decreases with a decrease in A1/A2, σci1/σci2 and σtm1/σtm2 and an increase in B1/B2, γ1/γ2. The range will decrease with a decrease in support pressure q and increase with a decrease in surface overload σ;s. Therefore, reinforced supporting is beneficial to improve the stability of the cavity during actual construction.

Based on the nonlinear Mohr-Coulomb failure criterion and the associated flow rules, the three-dimensional (3-D) axisymmetric failure mechanism of shallow horizontal circular plate anchors that are subjected to the ultimate pullout capacity (UPC) is determined. A derivative function of the projection function for projecting the 3-D axisymmetric failure surface on plane is deduced using the variation theory. By using difference principle, the primitive function of failure surface satisfying boundary condition and numerical solution to its corresponding ultimate pullout capacity function are obtained. The influences of nonlinear Mohr-Coulomb parameters on UPC and failure mechanism are studied. The result shows that UPC decreases with dimensionless parameter m and uniaxial tensile strength increases but increases when depth and radius of plate anchor, surface overload, initial cohesion, geomaterial density and friction angle increase. The failure surface is similar to a symmetrical spatial funnel, and its shape is mainly determined by dimensionless parameter m; the surface damage range expands with the increase of radius and depth of the plate anchor as well as initial cohesion but decreases with the increase of dimensionless parameter m and uniaxial tensile strength as well as geomaterial density. As the dimensionless parameter m=2.0, the numerical solution of UPC based on the difference principle is proved to be feasible and effective through the comparison with the exact solution. In addition, the comparison between solutions of UPC computed by variation method and those computed by upper bound method indicate that variation method outperforms upper bound method.

Crop improvement by domestication and traditional breeding often results in fitness penalties and loss of genetic diversity, which greatly threatens crop production and food security under the challenging global climate. De novo domestication has been proposed as a novel strategy for crop breeding. By combining multi-omics, genome editing and synthetic biology approaches, domestication of wild or semi-wild plant species can be accelerated by rapidly introducing desirable traits without causing an associated drag on their inherent traits. In this review, we summarize the history of crop domestication, emphasize the urgency for breeding strategy innovation, and discuss recent progress of de novo crop domestication.

Background The clinical and laboratory characteristics of AIDS-associated Talaromyces marneffei infection, a rare but a fatal mycosis disease of the central nervous system, remain unclear. Case presentation Herein, we conducted a retrospective study of ten AIDS patients with cerebrospinal fluid culture-confirmed central nervous system infection caused by Talaromyces marneffei. All 10 patients were promptly treated with antifungal treatment for a prolonged duration and early antiviral therapy (ART). Among them, seven patients were farmers. Nine patients were discharged after full recovery, while one patient died during hospitalization, resulting in a mortality rate of 10%. All patients initially presented symptoms and signs of an increase in intracranial pressure, mainly manifesting as headache, dizziness, vomiting, fever, decreased muscle strength, diplopia or even altered consciousness with seizures in severe patients. Nine patients (90%) showed lateral ventricle dilatation or intracranial infectious lesions on brain CT. Cerebrospinal fluid findings included elevated intracranial pressure, increased leukocyte count, low glucose, low chloride and high cerebrospinal fluid protein. The median CD4+ T count of patients was 104 cells/μL (IQR, 36–224 cells/μL) at the onset of the disease. The CD4+ T cell counts of three patients who eventually died were significantly lower (W = 6.00, p = 0.020) than those of the patients who survived. Conclusions The common clinical symptoms of T. marneffei central nervous system infection are associated with high intracranial pressure and intracranial infectious lesions. Earlier recognition and diagnosis and a prolonged course of amphotericin B treatment followed by itraconazole combined with early ART might reduce the mortality rate.

Novel representatives of the important group of biologically-active, dehydroabietic acid-bearing oxazolidinone moiety were synthesized to explore more efficacious and less toxic antitumor agents. Structures of all the newly target molecules were confirmed by IR, 1H-NMR, 13C-NMR, and HR-MS. The inhibitory activities of these compounds against different human cancer cell lines (MGC-803, CNE-2, SK-OV-3, NCI-H460) and human normal liver cell line LO2 were evaluated and compared with the commercial anticancer drug cisplatin, using standard MTT (methyl thiazolytetrazolium) assay in vitro. The pharmacological screening results revealed that most of the hybrids showed significantly improved antiproliferative activities over dehydroabietic acid and that some displayed better inhibitory activities compared to cisplatin. In particular, compound 4j exhibited promising cytotoxicity with IC50 values ranging from 3.82 to 17.76 &micro, M against all the test cell lines and displayed very weak cytotoxicity (IC50 &gt, 100 &micro, M) on normal cells, showing good selectivity between normal and malignant cells. Furthermore, the action mechanism of the representative compound 4j was preliminarily investigated by Annexin-V/PI dual staining, Hoechst 33258 staining, which indicated that the compound can induce cell apoptosis in MGC-803 cells in a dose-dependent manner and arrest the cell cycle in G1 phase. Therefore, 4j may be further exploited as a novel pharmacophore model for the development of anticancer agents.

Extracellular polymeric substances (EPSs) are biosynthetic polymers of microbial origin in the sludge activation process and crucially affect the properties of sludge in biological wastewater treatment reactors, such as the formation of sludge flocs, stabilization of sludge structure, and protection of microbes against noxious environmental conditions. However, the EPS extraction efficiency differs significantly according to the extraction method used. In this study, soluble EPSs and loosely bound EPSs can be extracted by centrifugation first and tightly bound EPSs in activated sludge require additional eight treatments for extraction, respectively. Three physical methods (centrifugation, sonication, and heating) and five chemical methods (cation exchange resin, NaOH, formaldehyde+NaOH, EDTA, and formaldehyde+EDTA) were tested, and the content and composition of TB-EPS were analyzed. Meanwhile, the functional groups and elements in TB-EPS were investigated. Results showed that the heating method did not introduce exogenous substances during the EPS extraction process and that the destruction of cells from this method was relatively slight. Heating was shown to be a gentle and efficient method in this study. The three-dimensional excitation and emission matrix (EEM) fluorescence spectroscopy showed that the cation exchange resin method had good extraction effect on humic-like and protein-like substances. As to fulvic-acid-like substances, NaOH was better than the other seven methods. Infrared spectroscopy showed that no notable difference appeared in the functional groups of the TB-EPS extracted by physical methods, whereas chemical methods induced big differences and showed particular bands that did not appear in the TB-EPS extracted by physical methods. Overall, the amounts of TB-EPS elements extracted using chemical methods were greater than those extracted using physical methods. In conclusion, a method must be selected and established for each case, taking into consideration the experimental purpose, and the most appropriate method should be chosen carefully.

Great attention has been paid to accelerate the start-up period and enhance floc properties and structural stability in activated sludge reactors with the aid of inorganic chemical agents such as calcium ion. The laboratory-scale sequencing batch reactors (SBRs) were operated continuously for 35 days to investigate the effect of calcium ion (Ca

Although constitutively activated nuclear factor-kappaB (NF-kappaB), attenuated transforming growth factor beta (TGFbeta) signaling, and TP53 mutations frequently occur in human cancers, how these pathways interact and together contribute to malignancy remains uncertain. Here, we found an association between overexpression of NF-kappaB-related genes, reduced expression of TGFbeta receptor (TbetaR) subunits and downstream targets, and TP53 genotype in head and neck squamous cell carcinoma (HNSCC). In response to recombinant TGFbeta1, both growth inhibition and TGFbeta target gene modulation were attenuated or absent in a panel of human HNSCC lines. However, in HNSCC cells that retained residual TGFbeta signaling, TGFbeta1 inhibited both constitutive and tumor necrosis factor alpha-stimulated NF-kappaB activity. Furthermore, HNSCC lines overexpressing mutant (mt) TP53 and human tumor specimens with positive TP53 nuclear staining exhibited reduced TbetaRII and knocking down mtTP53 induced TbetaRII, increasing TGFbeta downstream gene expression while inhibiting proinflammatory NF-kappaB target gene expression. Transfection of ectopic TbetaRII directly restored TGFbeta signaling while inhibiting inhibitor kappaBalpha degradation and suppressing serine-536 phosphorylation of NF-kappaB p65 and NF-kappaB transcriptional activation, linking these alterations. Finally, experiments with TbetaRII conditional knockout mice show that abrogation of TGFbeta signaling promotes the sustained induction of NF-kappaB and its proinflammatory target genes during HNSCC tumorigenesis and progression. Together, these findings elucidate a regulatory framework in which attenuated TGFbeta signaling promotes NF-kappaB activation and squamous epithelial malignancy in the setting of altered TP53 status., link_to_OA_fulltext

Purpose: To determine the nature and potential pharmacologic reversibility of deficient TP53 expression and function in head and neck squamous cell carcinomas (HNSCC) with wild-type TP53, previously associated with decreased sensitivity to cisplatin therapy. Experimental Design: TP53 genotype, mRNA and protein expression, TP53-induced p21 expression, and TP53 DNA–binding and reporter gene function were determined in a panel of nine previously characterized HNSCC cell lines from the University of Michigan squamous cell carcinoma (UM-SCC) series. The genotoxic drug doxorubicin and the anti-inflammatory and antimalarial drug quinacrine, previously identified as inducers of TP53, were used to examine the nature and potential reversibility of deficient TP53 expression and function. The specific role of inducible TP53 on function and cellular proliferation was confirmed using selective TP53 inhibitor pifithrin-α or short hairpin RNA knockdown. The capability of quinacrine to sensitize HNSCC to the cytotoxic effects of cisplatin was assessed. Results: UM-SCC cell lines with wild-type TP53 genotype underexpressed TP53 mRNA and protein when compared with normal human keratinocytes or UM-SCC with mutant TP53. Although doxorubicin failed to induce TP53 expression or functional activity, quinacrine induced TP53 mRNA and protein expression, increased TP53 reporter activity and p21 protein expression, and induced growth inhibition in these wild-type TP53 cell lines. Quinacrine-induced TP53 reporter activity and growth suppression were attenuated by pifithrin-α and TP53 short hairpin RNA knockdown. Furthermore, quinacrine sensitized UM-SCC to cisplatin in vitro. Conclusions: Deficient TP53 mRNA and protein expression underlies decreased function in a subset of HNSCC with wild-type TP53 and can be restored together with cisplatin sensitization by quinacrine.

Purpose: To determine if gene signatures differentially expressed in head and neck squamous cell carcinomas (HNSCC) are related to alterations in transcription factors nuclear factor-κB (NF-κB) and TP53 previously associated with decreased cell death, response to therapy, and worse prognosis. Experimental Design: Unique gene signatures expressed by HNSCC lines were identified by cDNA microarray, principal components, and cluster analyses and validated by quantitative reverse transcription-PCR (RT-PCR) and in situ hybridization. Bioinformatic analysis of the promoters and ontogeny of these clustered genes was done. Expression of proteins encoded by genes of a putative NF-κB signature, NF-κB p65, and TP53 were examined in HNSCC tissue specimens by immunostaining. Predicted promoter binding and modulation of expression of candidate NF-κB genes and cell survival were evaluated by p65 chromatin immunoprecipitation (ChIP) and small interfering RNA (siRNA) knockdown. Results: Two groups of HNSCC exhibiting distinct gene signatures were identified: cluster A enriched for histone genes, with a higher prevalence of TP53 promoter binding motifs; and cluster B enriched for injury response genes with NF-κB regulatory motifs. Coexpression of cluster B proteins was observed with strong NF-κB phospho-p65 and weak TP53 staining, and NF-κB phospho-p65 was inversely associated with TP53 (P = 0.02). Promoter binding of the NF-κB signature genes was confirmed by p65 ChIP, and down-modulation of their expression and cell death were induced by p65 siRNA. Conclusion: NF-κB promotes expression of a novel NF-κB–related gene signature and cell survival in HNSCC that weakly express TP53, a subset previously associated with inactivated wild-type TP53, greater resistance to chemoradiotherapy, and worse prognosis.

Rhamnolipid biosurfactants produced mainly by Pseudomonas sp. had been reported to possess a wide range of potential industrial application. These biosurfactants are produced as monorhamnolipid (MRL) and di-rhamnolipid (DRL) congeners. The present study deals with rhamnolipid biosurfactants produced by three bacterial isolates from crude oil. Biosurfactants produced by one of the strains (named as IMP67) was found to be very efficacious based on its critical micelle concentration (CMC) value and hydrocarbon emulsification property. Strikingly, antimicrobial and anti-biofilm potential of this biosurfactant were higher than biosurfactants produced by other two strains. Thin layer chromatography (TLC) analysis and rhamnose quantification showed that the rhamnolipids of IMP67 had more MRL congeners than biosurfactants of the other two strains. Emulsification and antimicrobial actions were affected by manual change of MRL and DRL congener proportions. Increase of MRL proportion enhanced emulsification index and antimicrobial property to Gram negative bacteria. This result indicated that the ratio of MRL and DRL affect the emulsification potentials of rhamnolipids, and suggested that high emulsification potentials might enhance rhamnolipids to penetrate the cell wall of Gram negative bacteria. In consistent, rhamnolipids of IMP67 reduced the MIC of some antibiotics against bacteria, suggesting the potential of biosurfactant as antibiotics synergist.

The preparation and characterization of some dipositive metal ion complexes derived from potassium 3-(pyridine-4-carbonylmethyl)-dithiocarbazate (PCDHK) are reported. The solid complexes of the composition ML.nH 2 O (M=Cu(II), Co(II), Mn(II), Zn(II), Cd(II), Ni(II), Pb(II), L = PCD - 2 , n = 0, 1, PCD - 2 = PCDHK-K + -H + ) and ML 2 .2H 2 O (M=UO 2 (IV), L=PCDH - 1 , PCDH - 1 =PCDHK-K + ) have been characterized by elemental analyses, IR, UV, and 1 H NMR spectra. The IR spectral data indicate that PCDHK behaves as either a mononegative or binegative ligand and coordinates in a tridentate or bridging tetradentate manner.

With the increasing use of engineered nanoparticles, these materials will inevitably be released into the environment with unknown consequences. The interactions between engineered nanoparticles and plants are of particular concern, and the uptake by plants may constitute one of the main routes of exposure for species at a higher trophic level, leading to possible biomagnification of nanoparticles in the food chain. A review of the current literature shows that nanoparticles can be taken up by plants and cause phytotoxicity. The mechanisms by which engineered nanoparticles penetrate plant cells are still not well understood. Most of the studies have been performed with a few plant species under non-natural conditions such as hydroponics, and have included only the germination or seedling growth stage. Nanoparticles may undergo biotransformation and subsequent translocation within plants, although the mechanisms are poorly understood. Phytotoxicity is an important aspect for assessing potential environmental impacts of nanoparticles, but the mode of toxicity remains to be investigated.

An acidophilic iron-sulfur oxidizing bacterium LY01 was isolated from acid mine drainage of coal in Guizhou Province, China. Strain LY01 was identified as Acidithiobacillusferrooxidans by morphological and physiological characteristics, and phylogenetic analysis of its 16S rRNA gene sequence. Strain LY01 was able to grow using ferrous ion (Fe2+), elemental sulfur (S0) and pyrite as sole energy source, respectively, but significant differences in oxidation efficiency and bacterial growth were observed when different energy source was used. When strain LY01 was cultured in 9K medium with 44.2 g x L(-1) FeSO4.7H2O as the substrate, the oxidation efficiency of Fe2+ was 100% in 30 h and the cell number of strain LY01 reached to 4.2 x 10(7) cell x mL(-1). When LY01 was cultured in 9K medium with 10 g x L(-1) S0 as the substrate, 6.7% S0 oxidation efficiency, 2001 mg x L(-1) SO4(2-) concentration and 8.9 x 10(7) cell x mL(-1) cell number were observed in 21 d respectively. When LY01 was cultured with 30 g x L(-1) pyrite as the substrate, the oxidation efficiency of pyrite, SO4(2-) concentration and cell number reached 10%, 4443 mg x L(-1) and 3.4 x 10(8) cell x mL(-1) respectively in 20 d. The effects of different heavy metals (Ni2+, Pb2+) on oxidation activity of strain LY01 cultured with pyrite were investigated. Results showed that the oxidation activity of strain LY01 was inhibited to a certain extent with the addition of Ni2+ at 10-100 mg x L(-1) to the medium, but the addition of 10-100 mg x L(-1) Pb2+ had no effect on LY01 activity.

We attempted to identify a locus for schizophrenia and related disorders in 24 nuclear families of schizophrenic probands using a predefined classification system for affected cases that included those disorders most clearly identified as sharing a genetic relationship with schizophrenia-schizoaffective disorder and schizotypal personality disorder. Initially, we evaluated 8 markers on chromosome 5 on the first 12 families with available genotyping and diagnostic assessments and, assuming autosomal dominant transmission, found a lod score of 2.67 for the D5S111 locus (5p14.1-13.1) in one large nuclear family (no. 17; sibship: n = 12; schizophrenia: n = 3; schizotypal personality disorder: n = 2); the other 11 families were much smaller, less complete, and provided little additional information. Other branches of no. 17 were then assessed and the 2-point lod score for family 17 rose to 3.72; using multipoint analysis the lod score in 17 was 4.37. When only schizophrenia was used to define affectedness, the positive evidence for linkage to D5S111 was greatly reduced. Sensitivity analysis indicated that the lod score is heavily dependent upon the predefined diagnostic criteria. Our studies of other families of schizophrenic probands eventually totalled 23, but linkage to D5S111 in these yielded a -2.41 lod score. Themore » results provide evidence for genetic linkage of the D5S111 locus to schizophrenia and related disorders in one family. It may be of interest that over several generations, almost all the ancestors of family 17 could be traced back to a small, relatively isolated, hill region of Puerto Rico. 74 refs., 2 figs., 2 tabs.« less

In this study, Acidithiobacillus ferrooxidans LX5 was cultured in 9K medium with pyrite and ferrous sulfate as the substrates. Results showed that the number of A. ferrooxidans LX5 acclimated by pyrite for 20 d was 3.0 x 10(7) cell/mL in the liquid, and the oxidation activity to pyrite was stronger. A. ferrooxidans LX5 cultured for 48 h was 1.0 x 10(8) cell/mL in medium with FeSO4. Extracellular polymeric substance (EPS) produced from A. ferrooxidans LX5 acclimated by pyrite was 2 395 microg/(10(10) cells) three times as much as that cultured with FeSO4. A high-sulfur coal desulfurization experiment were carried out with A. ferrooxidans LX5 domesticated respectively in medium with ferrous sulfate and pyrite. The results showed that the coal desulfurization rate was up to 72.4% after bioleaching for 13 d by A. ferrooxidans LX5 acclimated by pyrite, at the same time, the desulfurization rate was only 47.2% with A. ferrooxidans LX5 cultured with ferrous sulfate and reached 65.8% in 20 d. Therefore, the desulfurization efficiency can be improved of coal and shorten the desulfurization time when A. ferrooxidans LX5 acclimated with pyrite as the substrate.

Aerobic denitrifying performances of P. mendocina AD6 at various C/N ratios, dissolved oxygen (DO) concentrations and carbon sources including citrate, acetate, and glucose were evaluated through flask experiment. At an insufficient carbon concentration, namely C/N ratio of 3, total nitrogen (TN) removal efficiency was 42% by aerobic denitrification. When the C/N ratio was increased to 8, 15, and 23, the TN removal efficiency by AD6 quickly increased to 55%, 70%, and 69%, in which 7%, 20%, and 25% of TN losses could be contributed to the aerobic denitrification by AD6, respectively. Batch cultures were carried out in a series of 250 mL conical flasks containing 100, 50, and 25 mL of the nitrate-supplemented basal medium with C/N ratio of 15, DO of 7.15-8.08 mg/L and strain AD6. Flasks were shaken in a rotary shaker at 28 degrees C and 180 r/min to allow better gas exchange. DO concentration of culture medium declined rapidly to anoxia condition at the beginning of reaction followed by an aerobic level with above 2 mg/L of DO after 24 h reaction. Different carbon sources had strong influence on the aerobic denitrifying performance of strain AD6. The maximum consumption rates of citrate, acetate, and glucose by strain AD6 were 90%, 92%, and 41%, respectively. In the medium spiked with acetate as carbon source, TN removal efficiency and N losses due to aerobic denitrification increased to 14% and 5%, respectively, with comparison to that spiked with citrate. Evaluation on nitrogen removal performances of aerobic denitrifier should be cautious with flask experiment because aerobic, partial aerobic and total anoxic growing conditions could be changed by C/N ration and carbon source of medium.

Influence of Ca2+ on glucose-fed aerobic granule was investigated through sequencing batch reactor(SBR) and shaking flask experiments. Granules of two SBRs spiked with 30 mg/L and 100 mg/L of Ca2+ could be observed visually on day 20. In the first 50 days, the SBR II had higher mixed liquor suspended solid (MLSS) and lower sludge volume index (SVI) than those of SBR I. After 50 days, the difference of granule performance in two reactors gradually be reduced and little differences were found on day 80 or more, for example, MLSS of SBR I, II was 7.5 g/L and SVI arrived at 46-48 mL/g and 45-47 mL/g respectively. After 90 days, sludge Zeta potential of SBR I, II reached -12.5 - -12.6 mV. Furthermore, with influent of 0-200 mg/L Ca2+, sludge Zeta potential of shaking flask reached -14.4-14 after 50 days. Ca2+ concentration of influent had little influence on Zeta potential of sludge during granulation process. Microbiological observations confirmed some appreciable changes in microorganism population and diversities with the increase of Ca2+ concentration of influent. The role of Ca2+ in bioflocculation of granule can be mainly attributed to its cation bridging instead of charge neutralization. The elemental analysis of seed sludge and aerobic granule by X-ray fluorescence (XRF) showed that the content of Ca, Mg, K, and Na in granule were less than seed sludge expect for Fe, increasing 4.42% and 7.82% respectively. Divalent metal ions such as Ca2+ and Fe2+ were probably constituent of biopolymer. It was possible that the binding of divalent metal ion with extracellular polymeric substances (EPS) enhanced the granulation of seed sludge. Consequently, the pollutant removal efficiency of SBR II was higher than that of SBR I throughtout the trial. Both NH4+ -N and COD removal efficiencies reached 90% and both total N and total P removal efficiency were 65%-70% after day 70.

To investigate the features of B7-H1 expression on peripheral myeloid dendritic cells (mDCs) in patients with HIV infection and evaluate the correlations between B7-H1 expression and disease progression.Peripheral blood samples from 82 treatment-naïve patients with HIV infection, 28 viral complete responders (CRs) under antiretroviral therapy (ART) and 14 healthy controls (HCs) were collected. Flow cytometry was applied to investigate the expression of B7-H1 on mDCs and CD4 cell counts. Plasma HIV-1 viral load was detected by bDNA.The frequency of B7-H1 expression on mDCs were 14.15% +/- 2.63%, 3.31% +/- 0.51% and 0.52% +/- 0.10% in AIDS patients, asymptomatic HIV infected individuals and HCs respectively. As compared with HCs, B7-H1 was significantly up-regulated on mDCs in HIV/AIDS patients. The order was as follows: AIDS patientsasymptomatic HIV infected individualsHCs (all P0.05). Interestingly, the expression of B7-H1 on mDCs in long-term nonprogressors (LTNPs) was 3.12% +/- 1.14%. And it was lower than that in typical progressors (TPs) [8.12% +/- 1.37% (P = 0.001)]. Moreover, the expression of B7-H1 was negatively correlated with CD4 cell counts and positively correlated with plasma viral load in these patients (r = -0.631, P0.01 and r = 0.482, P0.01 respectively). The expression of B7-H1 on mDCs was significantly lower in ART complete responders than that in AIDS patients (6.59% +/- 1.43% vs 14.15% +/- 2.63%) (P0.01). Expression of B7-H1 on mDCs decreased markedly in patients whose CD4 cell counts greatly elevated after a successful antiretroviral treatment.The expression of B7-H1 on mDCs is significantly up-regulated in HIV/AIDS patients. With a close correlation with disease status, it acts as a marker of disease progression.

Using orthogonal experiment design, the total saponin constituents were obtained by refluxing extraction with alcohol and separated by macroporous adsorption resin and n-Butyl alcohol from the stem bark of Kalopanax septemlobus. According to the purity analysis and the yield, the extraction process was optimized. The results showed that the main saponin constituents were gained with a yield of 1.32% by using macroporous adsorption resin but 1.05% by using n-Butyl alcohol. The former was more efficient than the latter on both yield and color. The optimal process with isolation by macroporous adsorption resin is cheap, simple and practical.

Nitrogen removal efficiency was investigated in three subsurface flow constructed wetlands (CWs) with and without reed. Root bag made of nylon sieve with 300 mesh was used to enwrap the reed root in one of reed CWs to distinguish reed rhizosphere from non-rhizosphere. The CWs with root bag enwrapped reed root (hereinafter called as mesh CWs) and other CWs were fed with artificial ammonium-rich wastewater. The results indicated that the COD and N removal occurred mainly in the front of CWs, and C and nitrogen removal occurred concurrently along the stream way. When C/N ratio of influent was 5, the removal efficiencies of NH4+ -N in control CWs, reed CWs and mesh CWs were 66.2%, 94.2% and 82.2%, respectively. TN removal efficiencies were 67.2%, 90.7% and 76.1% respectively. Simultaneous nitrification and denitrification phenomenon in this study was also observed. The removal efficiency of organic carbon was different from nitrogen removal efficiency, mesh CWs showed the highest COD removal efficiency with 80.9%, while control CWs and reed CWs were 72.2% and 56.2%, respectively. C/N ratio of wastewater throughout the bed was more than 5 in three CWs, which indicated carbon source supply was enough for denitrification. The oxidation-reduction position (ORP) and concentration of total organic carbon in rhizosphere and non-rhizosphere were detected. The ORP in the front of mesh CWs's rhizosphere was much higher than that in control CWs and non-rhizosphere in mesh CWs, which were 11-311 mV and 62-261 mV, respectively. Root exudates also showed the difference between rhizosphere and non-rhizosphere in mesh CWs, the TOC of them were 21.3-54.6 mg x L(-1) and 6.65-12.0 mg x L(-1). Due to the higher ORP and concentration of TOC, the nitrogen removal efficiency in plant CWs was much higher than that in control CWs.

To find out the most suitable conditions for a whole body hyperthermia (WBH) model and the influence of these conditions on the blood brain barrier (BBB) disruption and brain edema in rats.Forty male Sprague-Dawley (SD) rats were randomly assigned to four groups (n=10 in each group): control group, group A, group B and group C. After anesthesia with pentobarbital, rats were subjected to femoral artery and vein cannulation. Rats of control group were housed at a controlled room temperature (25-26 degrees C) for 4 hours. Rats of group A, group B and group C were exposed to WBH in a biological oxygen supply heated container (relative humidity 65%, wind velocity 25 cm/s) maintained at 34, 36 and 38 degrees C for 3 hours, respectively. Then the rats were removed from the heated container and their body temperature was cooled down for 1 hour. During heating, rectal temperature, heart rate (HR), mean arterial pressure (MAP), pH, partial pressure of oxygen in artery (PaO(2)), partial pressure of carbon dioxide in artery (PaCO(2)), the dosage of anesthetic, and the mortality rate in each group were recorded. Evans blue (EB) was administered into the femoral vein and allowed to circulate for 5 minutes. At the end of the experiment, the animals were perfused with 0.9% saline and heparin through the heart, and the brain was harvested for the examination of BBB permeability, water content and morphological alterations in brain tissues and neurons.The total dosage of pentobarbital was not significantly different among all groups. After WBH for 3 hours, the average rectal temperature was higher than rats without WBH, and the mortality rate was 0, 10%, 10% and 40% in groups control, A, B, C, respectively. HR of groups A, B and C were significantly higher than those of control group; MAP, pH of group A, B and C were significantly lower than those of control group (all P0.05). Compared to that of control group, water content of the brain and permeability of EB in groups A, B and C were significantly increased (P0.05 or P0.01), but there was no marked difference on PaO(2), PaCO(2) and haematocrit (HCT) among groups A, B and C. Morphological investigation showed that there were different degrees of structural changes in brain tissue in groups A, B and C under light microscopy. Under transmission election microscopy, the structure of nerve cells and BBB in group B and group C showed moderate to profound alterations, but there were no changes in group A.Rats housed in a biological oxygen supply heat container with the temperature maintained at 36 degrees C for 3 hours could establish an ideal WBH model with notable BBB breakdown, moderate brain edema, and histological changes in brain.

A subsurface-horizontal flow constructed wetlands (CWs) planted with reed was used to treat micro-polluted river water in this study with an aim to investigate the long-term treatment efficiency of CWs especially for organic C and N. Average data obtained from two-year plant growth season showed that performance of the wetlands appeared to be affected by both establishment/maturation factors and year-to-year climatic variations. The results displayed that the removal of C and N in the influent depended, to a certain extend, on plant growth and seasonal variations, especially for total N removal. It was observed that C removal occurred mainly in the front of CWs in the first-year's operation period and then was translocated to the rear end of wetlands in the second-year's operation period. C/N ratio in the influent was 5 or more, indicating enough C source supply for denitrification. Organic C removal efficiencies varied from 6.10% to 37.83% throughout the trial. Average total N removal efficiency of 15.51% in the first-year operation period and then declined to 8.61% in the second year. The highest removal efficiency of total N was below 40% throughout the two-year trial. It was found that nitrification and denitrification reached dynamic equilibrium at the middle of the wetlands where the highest total N removal efficiency occurred. The greatest oxygen consumption was observed in the front and middle of CWs. It was noted that nitrification occurred even in deep layer located in the rear end of the wetlands in the second-year operation period. Nitrification and denitrification occurred concurrently with C and total N removal along the stream way. Low-molecular-weight organic acids released from reed rhizosphere seemed to have a significant inhibitory effect on chemoautrophic nitrifying bacteria, which involved in nitrogen removal efficiency of the wetlands, particularly during spring and autumn.

Squamous cell carcinomas and several other cancers have been found to exhibit microarray expression profiles that include genes related to nuclear factor (NF)-kappaB, a signal activated transcription factor that is evolutionarily important in regulating early response gene programs to injury and infection. Inhibition of NF-kappaB by expression of a dominant negative signal phosphorylation site mutant of inhibitor-kappaB, IkappaBalphaM, under a tetracycline inducible promoter, established the role of NF-kappaB as an essential molecular switch modulating multiple genes important in the malignant phenotype. Bioinfomatic analysis of the promoter and coding region of IkappaBalphaM-modulated genes has enabled identification of new candidates with and without known NF-kappaB related motifs for validation and functional studies of their relationship to NF-kappaB. These studies illustrate how microarray data can be used to generate a hypothesis regarding regulation of genes by a specific signal transcription factor, and how genetic mutants and bioinformatic analysis can be used to analyze the relative importance of the regulatory molecule to expression of genes involved in the malignant phenotype.

To determine the effect of ulinastatin (UTI) on blocking the development of acute lung injury (ALI) induced by acid aspiration in the early period.The SD rats were randomly divided into 3 groups: control group (n = 10), injury group (n = 10), and UTI-treated group (n = 10). The rats in the control group were injected intrabronchialy saline while the other 2 groups were injected HCl. The UTI-treated group was injected UTI at 4 x 10(4) U/kg after acid instillation. Blood gas as well as wet/dry weight ratio (W/D) of the left lung was measured. The changes of IL-8 and TNF-alpha levels in the pulmonary tissues were determined by enzyme-linked immunosorbent assay.The result of blood gas analysis in the UTI-treated group was much better than that in the injury group (P0.01). W/ D and IL-8, TNF-alpha levels were lower in the UTI-treated group than those in the injury group significantly (P0. 01), and there was no difference between the UTI-treated group and the control group in IL-8 and TNF-alpha levels (P0. 05).UTI administration can lessen the development of ALI induced by acid aspiration in the early period.

Hepatocyte growth factor/scatter factor (HGF) and the angiogenesis factors platelet-derived growth factors (PDGF), vascular endothelial growth factor (VEGF), and interleukin-8 (IL-8) are found in elevated concentrations in serum or tumor tissue of patients with head and neck squamous cell carcinomas (HNSCC), suggesting these factors may be coregulated. A cDNA microarray analysis for HGF-inducible genes revealed that HGF also modulates PDGFA expression, a gene recently shown to be inducible by the transcription factor, early growth response-1 (Egr-1). In the present study, we investigated the potential role of HGF-induced Egr-1 in expression of PDGF, VEGF, and IL-8. HGF induced expression of all three factors and Egr-1 expression and DNA-binding activity. The analysis of promoter sequences showed putative Egr-1 binding sites in the PDGFA or VEGF but not in the IL-8 promoter, and HGF-induced Egr-1–binding activity was confirmed by chromatin immunoprecipitation (ChIP) assay. The maximal basal and HGF-induced promoter activity for the PDGFA gene existed within −630 bp of the promoter region, and overexpression of Egr-1 significantly increased such activity. Consistent with this, expression of PDGFA and VEGF but not IL-8 showed corresponding differences with Egr-1 expression in HNSCC tumor specimens and were strongly suppressed by transfection of Egr-1–antisense or small interference RNA (siRNA) oligonucleotides. HGF-induced expression of Egr-1, PDGFA, and VEGF was suppressed by pharmacologic and siRNA inhibitors of mitogen-activated protein kinase kinase 1/2 (MEK1/2) and protein kinase C (PKC) pathways. We conclude that the HGF-induced activation of transcription factor Egr-1 by MEK1/2- and PKC-dependent mechanisms differentially contributes to expression of PDGF and VEGF, which are important angiogenesis factors and targets for HNSCC therapy.

To investigate the changes in transcription and expression of inducible nitric oxide synthase (NOS2) in the liver ischemic preconditioning (IP), and to determine the role of nitric oxide (NO) synthetic pathway in the liver IP in rats.We randomly divided 131 Sprague Dawley rats into 3 groups: ischemia/reperfusion (I/R) group (n=52), IP group (n=41), and sham operation (S) group (n=38). Plasm NO concentration and the transcription and expression of NOS2 were detected 2 hours, 24 hours, and 1 week after the operation.(1) In the IP group, the NO concentrations at the 2nd hour, the 24th hour, and 1 week were significantly higher than those in the S group (P0.05, P0.01, P0.01, respectively) and the NO concentrations at the 2nd hour and the 24th hour were obviously higher than those in I/R group (all P0.01). In the I/R group, the NO concentration at the 2nd hour was significantly lower than that in the S group (P0.05); there was no significant difference between the I/R group and the S group 24 hours after the operation (P0.05), and the NO concentration 1 week after the operation was obviously higher than that in the S group (P0.05). (2) In the IP group, the transcription of NOS2 2 and 24 hours after the operation were significantly increased compared with that in the I/R group (all P0.05) , but after 1 week, the transcription was not statistically different between the IP group and the I/R group (P0.05). (3) In the IP group, the expressions of NOS2 after 2 and 24 hours were obviously higher than those in the I/R group (P0.05) and the S group (P0.05), but the expression between the IR group and the S group was not significantly different (P0.05); after 1 week, the expressions of NOS2 in IP group and I/R group were weakly positive (P0.05) , and those in S group were negative (P0.05). Conclusion The transcription and the expression of liver NOS2 increase after the liver received ischemic preconditioning in rats. That the peak phases of transcription and expression of NOS2 are ahead of time may be related to the early protective effect of the liver IP.

Purpose: Head and neck squamous cell carcinomas have been reported to overexpress hypoxia-inducible factor (HIF)-1α, a transcription factor that promotes expression of angiogenesis factors and resistance to programmed and therapy-induced cell death. 2-Methoxyestradiol (2ME2) is a natural compound with HIF-1α inhibitory activity that is currently being evaluated in phase 1 and 2 clinical trials for advanced solid tumors and multiple myeloma. To our knowledge, this is the first study to evaluate the effects of 2ME2 in head and neck squamous cell carcinoma. Experimental Design: In the present study, we investigated the effects of 2ME2 alone and in combination with paclitaxel, an active agent in recurrent or advanced head and neck squamous cell carcinoma. Results: 2ME2 exhibited antiproliferative and cytotoxic effects in a panel of five head and neck squamous cell carcinoma cell lines in the 0.5 to 10 μmol/L range, including induction of G2-M blockade, caspase-3/7 activation, and apoptosis at 48 hours. 2ME2 resulted in decreased nuclear HIF-1α–binding activity and affected the expression of downstream genes, such as bid, a proapoptotic bcl-2 family member, and vascular endothelial growth factor, a proangiogenic cytokine. The up-regulation of Bid (57.5% at 12 hours, P < 0.0006) and inhibition of vascular endothelial growth factor secretion (57.7% at 24 hours, P < 0.015; and 50.3% at 48 hours, P < 0.0006) could be partially attributed to the effects on HIF-1α, because HIF-1α small interfering RNAs produced similar effects. Finally, in vivo, in a xenograft model of head and neck squamous cell carcinoma using UM-SCC-11A cells, 2ME2 exhibited antitumor and antiangiogenic activity, as measured by CD31 immunostaining. Conclusions: These results provide support for the use of 2ME2 in combination with paclitaxel for the treatment of recurrent or advanced head and neck squamous cell carcinoma.