Gene transfer of CD40 ligand (CD40L) holds promise as a novel therapy for lymphoid malignancies and a number of solid carcinomas because of its multiple anti-tumor activities. However, membrane-bound CD40L can be cleaved into a soluble form, sCD40L, which contributes to systemic inflammatory and cardiovascular diseases, and induces survival signals in the absence of protein synthesis block, suggesting a deleterious side effect of CD40L gene therapy. We generated a plasmid encoding non-cleavable human CD40L mutant (pcDNA3.1+-CD40L-M) to determine the direct anti-proliferative and pro-apoptotic effects in CD40-positive lung adenocarcinoma cell line A549, to verify activation of immature dentritic cells (DCs) by co-cultivation with the transfected A549 cells and to evaluate the lower expression of sCD40L relative to that of wild-type CD40L (CD40L-WT) transfectant in cell-free supernatants. These studies suggest that gene transfer of the membrane-stable CD40L mutant into CD40-positive cells may provide an efficient and safe method to treat non-small cell lung cancer.

Gene C4orf7, renamed FDC-SP (follicular dendritic cell secreted protein, FDC-SP) was first isolated from human tonsils. Up to the present, the function of this gene was still poorly understood. In this study, we report the expression of gene C4orf7 in a panel of tumor types. The percentages of C4orf7 positive expression in the tumor patients with metastases were notably higher than those in the cancer without metastases. On the contrary, the expression was hardly noted in normal tissues and corresponding benign lesions. In vitro, the up-regulation of C4orf7 in ovarian cancer cell lines was associated with enhanced motility and invasiveness. Furthermore, the overexpression of C4orf7 resulted in Akt ser473 phosphorylation and decreased E-cadherin expression. The role of C4orf7 in ovarian cancer cell morphology, motility and invasion was demonstrated.

Spinal muscular atrophy is the most common fatal hereditary disease among newborns and infants. There is as yet no effective treatment. Although a carrier test is available, currently there is disagreement among professional medical societies who proffer standards of care as to whether or not carrier screening for spinal muscular atrophy should be offered as part of routine reproductive care. This leaves health care providers without clear guidance. In fall 2009, a meeting was held by National Institutes of Health to examine the scientific basis for spinal muscular atrophy carrier screening and to consider the issues that accompany such screening. In this article, the meeting participants summarize the discussions and conclude that pan-ethnic carrier screening for spinal muscular atrophy is technically feasible and that the specific study of implementing a spinal muscular atrophy carrier screening program raises broader issues about determining the scope and specifics of carrier screening in general.

PURPOSE OF REVIEW: Condoms and vasectomy are male-controlled family planning methods but suffer from limitations in compliance (condoms) and limited reversibility (vasectomy); thus many couples desire other options. Hormonal male contraceptive methods have undergone extensive clinical trials in healthy men and shown to be efficacious, reversible and appear to be well tolerated. RECENT FINDINGS: The success rate of male hormonal contraception using injectable testosterone alone is high and comparable to methods for women. Addition of progestins to androgens improved the rate of suppression of spermatogenesis. Supported by government or nongovernment organizations, current studies aim to find the best combination of testosterone and progestins for effective spermatogenesis suppression and to explore other delivery methods for these hormones. Translation of these advances to widespread use in the developed world will need the manufacturing and marketing skills of the pharmaceutical industry. Availability of male contraceptives to the developing world may require commitments of governmental and nongovernmental agencies. In a time when imbalance of basic resources and population needs are obvious, this may prove to be a very wise investment. SUMMARY: Male hormonal contraception is efficacious, reversible and well tolerated for the target population of younger men in stable relationships. Suppression of spermatogenesis is achieved with a combination of an androgen and a progestin. Partnership with industry will accelerate the marketing of a male hormonal contraceptive. Research is ongoing on selective androgen and progesterone receptor modulators that suppress spermatogenesis, minimize potential adverse events while retaining the androgenic and gonadotropin suppressive actions.

Objective-To evaluate the effects of various flow rates of oxygen administered via 1 or 2 nasal cannulae on the fraction of inspired oxygen concentration (Fio(2)) and other arterial blood gas variables in healthy neonatal foals. Animals-9 healthy neonatal (3- to 4-day-old) foals. Procedures-In each foal, a nasal cannula was introduced into each naris and passed into the nasopharynx to the level of the medial canthus of each eye; oxygen was administered at 4 flow rates through either 1 or both cannulae (8 treatments/foal). Intratracheal Fio(2), intratracheal end-tidal partial pressure of carbon dioxide, and arterial blood gas variables were measured before (baseline) and during unilateral and bilateral nasopharyngeal delivery of 50, 100, 150, and 200 mL of oxygen/kg/min. Results-No adverse reactions were associated with administration of supplemental oxygen except at the highest flow rate, at which the foals became agitated. At individual flow rates, significant and dose-dependent increases in Fio(2), Pao(2), and oxygen saturation of hemoglobin (Sao(2)) were detected, compared with baseline values. Comparison of unilateral and bilateral delivery of oxygen at similar cumulative flow rates revealed no differences in evaluated variables. Conclusions and Clinical Relevance-Results indicated that administration of supplemental oxygen via nasal cannulae appeared to be a highly effective means of increasing Fio(2), Pao(2), and Sao(2) in neonatal foals. These findings may provide guidance for implementation of oxygen treatment in hypoxemic neonatal foals. (Am J Vet Med 2010;71:1081-1088).

Protein-protein docking has made much progress in recent years, but challenges still exist. Here we present the application of our docking approach HoDock in CAPRI. In this approach, a binding site prediction is implemented to reduce docking sampling space and filter out unreasonable docked structures, and a network-based enhanced combinatorial scoring function HPNCscore is used to evaluate the decoys. The experimental information was combined with the predicted binding site to pick out the most likely key binding site residues. We applied the HoDock method in the recent rounds of the CAPRI experiments, and got good results as predictors on targets 39, 40, and 41. We also got good results as scorers on targets 35, 37, 40, and 41. This indicates that our docking approach can contribute to the progress of protein-protein docking methods and to the understanding of the mechanism of protein-protein interactions. Proteins 2010. (c) 2010 Wiley-Liss, Inc.

Plant calcium-dependent protein kinases (CDPKs) may function as calcium sensors and play important roles in regulation of plant growth and development and in plant responses to biotic and abiotic stresses. The Arabidopsis genome encodes 34 CDPKs and most of them have not been functionally characterized. Here we report the functional characterization of CPK10 in Arabidopsis response to drought stress. The cpk10 mutant, a T-DNA insertion mutant for Arabidopsis CPK10 gene, showed much more sensitive phenotype to drought stress compared with wild-type plants, while the CPK10 overexpression lines displayed enhanced tolerance to drought stress. Induction of stomatal closure and inhibition of stomatal opening by ABA and Ca(2+) were impaired in the cpk10 mutants. Using the yeast two-hybrid methods, a heat shock protein HSP1 was identified as a CPK10-interacting protein. The interaction between CPK10 and HSP1 was further confirmed by pull-down and bimolecular fluorescence complementation (BiFC) assays. The HSP1 knock-out mutant (hsp1) plants showed similar sensitive phenotype under drought stress as the cpk10 mutant plants, and were similarly less sensitive to ABA and Ca(2+) in regulation of stomatal movements. Electrophysiological experiments showed that ABA- and Ca(2+)-inhibition of the inward K(+) currents in stomatal guard cells were impaired in the cpk10 and hsp1 mutants. All presented data demonstrated that CPK10, possibly via interacting with HSP1, plays important roles in ABA and Ca(2+)-mediated regulation of stomatal movements.

The purpose of this study was to investigate the potentiation of the anticancer activity and enhanced cellular retention of paclitaxel-loaded PLGA nanoparticles after surface conjugation with wheat germ agglutinin (WGA) against colon cancer cells. Glycosylation patterns of representative colon cancer cells confirmed the higher expression levels of WGA-binding glycoproteins in the Caco-2 and HT-29 cells, than in the CCD-18Co cells. Cellular uptake and in vitro cytotoxicity of WNP (final formulation) against colon cell lines was evaluated alongside control formulations. Confocal microscopy and quantitative analysis of intracellular paclitaxel were used to monitor the endocytosis and retention of nanoparticles inside the cells. WNP showed enhanced anti-proliferative activity against Caco-2 and HT-29 cells compared to corresponding nanoparticles without WGA conjugation (PNP). The greater efficacy of WNP was associated with higher cellular uptake and sustained intracellular retention of paclitaxel, which in turn was attributed to the over-expression of N-acetyl-D-glucosamine-containing glycoprotein on the colon cell membrane. WNP also demonstrated increased intracellular retention in the Caco-2 (30% of uptake) and HT-29 (40% of uptake) cells, following post-uptake incubation with fresh medium, compared to the unconjugated PNP nanoparticles (18% in Caco-2) and (27% in HT-29), respectively. Cellular trafficking study of WNP showed endocytosed WNP could successful escape from the endo-lysosome compartment and release into the cytosol with increasing incubation time. It may be concluded that WNP has the potential to be applied as a targeted delivery platform for paclitaxel in the treatment of colon cancer.

To understand the tail-suspension model to simulate weightlessness better, this study was to investigate the relationship of the amount of body weight supported by forelimbs between the tilt angles of rat in the model. Normal rat had at least two basic postures. One was standing or walking, in which the forelimbs bear 44.6% of the body weight; the other one was resting, in which 23.9% of body weight was placed on the forelimbs. As for tail-suspended rat, body-weight distribution on forelimbs was linearly related to tilt angle. The linear relationship was y = -0.7423x + 70.849, R2 = 0.9269. The tilt angle should be approximately 35 degrees if normal standing load of 44.6% body weight was placed on the forelimbs. On the other hand, it should be approximately 63 degrees if normal resting load of 23.9% of body weight was placed on forelimbs. Furthermore, the body load on forelimbs in tail-suspension model became much larger if the period of different postures was considered. Therefore, it should be careful if forelimbs are used to be as convenient internal control in tail-suspended rats.

Increasing evidence suggests that the presence of endotoxemia is of substantial clinical relevance to patients with cirrhosis, but it is unclear whether and how gut-derived LPS amplifies the tumorigenic response of the liver. We found that the circulating levels of LPS were elevated in animal models of carcinogen-induced hepatocarcinogenesis. Reduction of LPS using antibiotics regimen in rats or genetic ablation of its receptor Toll-like receptor 4 (TLR4) in mice prevented excessive tumor growth and multiplicity. Additional investigation revealed that TLR4 ablation sensitizes the liver to carcinogen-induced toxicity via blocking NF-kappaB activation and sensitizing the liver to reactive oxygen species (ROS)-induced toxicity, but lessens inflammation-mediated compensatory proliferation. Reconstitution of TLR4-expressing myeloid cells in TLR4-deficient mice restored diethylnitrosamine (DEN)-induced hepatic inflammation and proliferation, indicating a paracrine mechanism of LPS in tumor promotion. Meanwhile, deletion of gut-derived endotoxin suppressed DEN-induced cytokine production and compensatory proliferation, whereas in vivo LPS pre-challenge promotes hepatocyte proliferation. Conclusion: Our data indicate that sustained LPS accumulation represents a pathological mediator of inflammation-associated hepatocellular carcinoma (HCC) and manipulation of the gut flora to prevent pathogenic bacterial translocation and endotoxin absorption may favorably influence liver function in patients with cirrhosis who are at risk of developing HCC. (HEPATOLOGY 2010.).

Mutations in the parkin gene have been identified as one of the important genetic factors for the etiology of PD. However, pathogenicities of parkin mutations, especially of those heterozygotes, remain controversial, possibly due to confusions caused by the "mixed" effects of all types of mutations. Here we report a study on the independent effects of exonic deletions/duplications (or dosage mutations) on the risk for and clinical profiles of PD in a Chinese cohort consisting of 29 autosomal recessive early-onset Parkinsonism(AREP) families and 173 patients with sporadic early-onset Parkinson's disease (EOPD).Detected simultaneously by multiplex PCR/denaturing high performance liquid chromatography and real-time quantitative PCR analysis, heterozygous or homozygous dosage mutations were identified in exons 2-7.of parkin. The overall frequency of these mutations for both types of EOPD patients (8.91%, 18/202) was significantly higher than that of the controls (0%, 0/54) (p=0.03), and heterozygous mutations were more frequent in the AREP probands (17.2%,5/29) than in the controls (0%) (p=0.004) and the sporadic EOPD cases (3.5%, 6/173) (p=0.01). AREP patients with dosage mutations had significantly younger age at onset (AAO) (p=0.000) and were more likely to present as bradykinesia and complicated with dystonia or dyskinesia compared with those without mutations. Sporadic EOPD patients with dosage mutations do not have an earlier AAO, but are more likely to present as tremor and bradykinesia. These data suggested that dosage mutations alone can increase the risk for both sporadic and familial EOPD and affect their clinical aspects, but might contribute to a greater degree to familial EOPD.

BACKGROUND:: Stent graft-induced new entry (SINE), defined as the new tear caused by the stent graft and excluding those arising from natural disease progression or iatrogenic injury from the endovascular manipulation, has been increasingly observed after thoracic endovascular aortic repair (TEVAR) for Stanford type B dissection in our center. SINE appears to be remarkably life threatening. We investigated the incidence, mortality, causes, and preventions of SINE after TEVAR for Stanford type B dissection. METHODS:: Data for 22 patients with SINE were retrospectively collected and analyzed from 650 patients undergoing TEVAR for type B dissection from August 2000 to June 2008. An additional patient was referred to our center 14 months after TEVAR was performed in another hospital. The potential associations of SINE with Marfan syndrome, location of SINE and endograft placement, and the oversizing rate were analyzed by Fisher exact probability test or t test. RESULTS:: We found 24 SINE tears in 23 patients, including SINE at the proximal end of the endograft in 15, at the distal end in 7, and at both ends in 1. Six patients died. SINE incidence and mortality reached 3.4% and 26.1%, respectively. Two SINE patients were diagnosed with Marfan syndrome, whereas there were only 6 Marfan patients among the 651 patients. The 16 proximal SINEs were evidenced at the greater curve of the arch and caused retrograde type A dissection. The eight distal SINEs occurred at the dissected flap, and five caused enlarging aneurysm whereas three remained stable. The endograft was placed across the distal aortic arch during the primary TEVAR in all 23 patients. The incidence of SINE was 33.33% among Marfan patients vs 3.26% among non-Marfan patients (P = .016). There was no significant difference in mortality between proximal and distal SINE (25% vs 28.6%, P > .99), incidence of SINE between endograft placement across the arch and at the straight portion of descending thoracic aorta (23 of 613 vs 0 of 38, P = .39), and the oversizing rate between SINE and non-SINE patients (13% +/- 4.5% vs 16% +/- 6.5%, P = .98). CONCLUSIONS:: SINE appears not to be rare after TEVAR for type B dissection and is associated with substantial mortality. The stress yielded by the endograft seems to play a predominant role in its occurrence. It is important to take this stress-induced injury into account during both design and placement of the endograft.

A fiber confocal backscattering (FCBS) spectrometer is developed based on fiber confocal microscopy and light scattering theory. The FCBS spectrometer can provide imaging and spectral information simultaneously at the cellular scale. Normal stomach epithelial cell line GES-1 and cancerous cell line NCI-N87 are measured and their spectral results show that backscattering intensity from NCI-N87 cells is stronger than that from GES-1 cells in 500 to 800 nm, and the GES-1 cells scattering spectra show regular intensity changes, while the NCI-N87 cells do not. The experiments prove that the FCBS spectrometer is able to distinguish cancerous cells from normal stomach cells at the cellular level. The spectrometer could be further developed into a noninvasive optical technology for early cancer detection.

We studied throat swabs and corresponding serum samples collected from 1067 protein purified derivative (PPD)-tuberculin skin test (TST) positive cattle from different regions of China. The 1067 throat swabs were inoculated onto modified Löwenstein-Jensen medium for the isolation and culture of Mycobacteria. Acid-fast bacilli were identified using traditional biochemical methods, polymerase chain reaction (PCR) amplification and multiplex PCR. They were distinguished as Mycobacterium tuberculosis complex (MTBC) and non-tuberculous mycobacteria (NTM) strains. An indirect Enzyme-Linked Immunosorbent Assay (ELISA) was applied to detect specific antibodies against bovine TB (bTB). Correlations among the ELISA, bacteriology and TST were analyzed and compared. Spoligotyping and variable number tandem repeats-mycobacterial interspersed repetitive unit (VNTR-MIRU) analysis were used to genotype the MTBC. In total, 111 strains of Mycobacteria were cultured from the 1067 throat swab samples, including 43 stains of MTBC (14 strains of Mycobacterium bovis and 29 of Mycobacterium tuberculosis) and 68 strains of NTM. Thirty-eight MTBC strains and four NTM strains were isolated from 72 throat swab samples that the ELISA determined were antibody positive; five MTBC strains and 64 NTM strains were isolated from 995 throat swab samples that were antibody negative on the ELISA. The positive isolation rates of MTBC and NTM were 38.7% (43/111) and 61.3% (68/111), respectively. The concordance rate of cultured MTBC with a positive result on the indirect ELISA for antibody was 52.8% (38/72), which was much higher than the positive rate for TST (4.0%; 43/1067). Genotyping of the 43 strains of MTBC isolated, using spoligotyping and VNTR-MIRU, showed that the 43 isolates had 26 genotypes; 16 strains had a unique genotype. Two groups of six strains and two strains, respectively, showed the same spoligotyping pattern, and belonged to the Beijing family and Beijing-like family, respectively. Combined application of spoligotyping and VNTR-MIRU typing would improve the molecular epidemiological investigation and monitoring of the etiology of bTB in China.

Although the beneficial effects of n-3 fatty acids on several physiologic functions have been widely reported, information about the effects of oily fish in the Asian diet on cardiovascular disease (CVD) risk is diminutive. We hypothesize that daily inclusion of oily fish for 8 weeks in the Chinese diet will elevate serum eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels and reduce CVD risk markers in dyslipidemic adult men, comparable with the effects generally observed by inclusion of oily fish in the Western diet. In this 8-week randomized, parallel-arm, food-based intervention study, lunches were prepared with 500 g pork/chicken/beef, typically consumed fish (hairtail and freshwater carp), or oily fish (salmon). Male subjects aged between 35 and 70 years with hyperlipidemia were randomly assigned to eat lunches with pork/chicken/beef (n = 30 subjects at 8 weeks), freshwater fish (n = 30), or oily fish (n = 32). Circulating markers were measured at baseline and at 8 weeks. In the oily fish diet, dietary EPA and DHA levels were significantly increased as compared with other diets; and the n-6:n-3 polyunsaturated fatty acid ratio was decreased (P < .05). Thus, the oily fish diet significantly elevated serum EPA and DHA concentrations (P < .01) and lowered serum n-6:n-3 ratio at 8 weeks (P < .05). Furthermore, oily fish intake significantly reduced serum levels of triglycerides (P < .05) and interleukin-6 (P < .01) and increased levels of high-density lipoprotein cholesterol (P < .01). In conclusion, daily inclusion of oily fish as part of the Chinese diet for 8 weeks is sufficient to significantly increase the serum content of long-chain n-3 polyunsaturated fatty acids and reduce levels of CVD risk markers in dyslipidemic adult men.

Abstract Aims: To characterize the bacterial communities in commercial total mixed ration (TMR) silage, which is known to have a long bunk life after silo opening. Methods and Results: Samples were collected from four factories that produce TMR silage according to their own recipes. Three factories were sampled three times at 1-month intervals during the summer to characterize the differences between factories; one factory was sampled 12 times, three samples each during the summer, autumn, winter and spring, to determine seasonal changes. Bacterial communities were determined by culture-independent denaturing gradient gel electrophoresis. All silages contained lactic acid as the predominant acid, and the contents appeared stable regardless of factories and product seasons. Acetic acid and 1-propanol contents were different between factories and indicated seasonal changes, with increases in warm seasons compared to cool seasons. Both differences and similarities existed among the bacterial communities from each factory and product season. Lactobacillus parabuchneri was found in the products from three of four factories. Various sourdough lactic acid bacteria (LAB) were identified in commercial TMR silage; Lactobacillus panis, Lactobacillus hammesii, Lactobacillus mindensis, Lactobacillus pontis, Lactobacillus frumenti and Lactobacillus farciminis were detected in many products. Moreover, changes owing to product season were distinctive, and Lact. pontis and Lact. frumenti became detectable in summer products. Conclusion: Sourdough LAB are involved in the ensiling of commercial TMR silage. Silage bacterial communities vary more by season than by factory. The LAB species Lact. parabuchneri was detected in the TMR silage but may not be essential to the product's long bunk life after silo opening. Significance and Impact of the Study: Commercial TMR silage resembles sourdough with respect to bacterial communities and long shelf life. The roles of sourdough LAB in the ensiling process and aerobic stability are worth examining.

Abstract This work investigated the cytotoxicities of three silver nanoparticles (SNPs) SNP-5, SNP-20 and SNP-50 with different sizes ( approximately 5 nm, approximately 20 nm and approximately 50 nm) using four human cell models (A549, SGC-7901, HepG2 and MCF-7). Endpoints included cell morphology, cell viability, cellular membrane integrity, oxidative stress and cell cycle progression. Observable deleterious effects on the cell morphologies and membrane integrity were induced by SNP-5 and SNP-20. SNPs elevated the ROS levels in cells and arrested the cells at S phase. Apoptosis occurred for 4-9% of the exposed cells. All these cellular responses as well as EC50 values were found to be size-dependent for the tested SNPs. Ultrastructural observations confirmed the presence of SNPs inside cells. Elemental analysis of silver in cells by ICP-MS showed that smaller nanoparticles enter cells more easily than larger ones, which may be the cause of higher toxic effects. The findings may assist in the design of SNP applications and provide insights into their toxicity.

Fusarium head blight or scab caused by Fusarium graminearum is an important disease of wheat and barley. The pathogen not only causes severe yield losses but also contaminates infested grains with mycotoxins. In a previous study we identified several pathogenicity mutants by random insertional mutagenesis. One of these mutants was disrupted in the ZIF1 gene, which encodes a b-ZIP transcription factor unique to filamentous ascomycetes. The Deltazif1 mutant generated by gene replacement was significantly reduced in deoxynivalenol (DON) production and virulence on flowering wheat heads. It was defective in spreading from inoculated florets to the rachis and other spikelets. Deletion of the ZIF1 ortholog MoZIF1 in the rice blast fungus also caused reductions in virulence and in invasive growth. In addition, the Deltazif1 mutant is defective in sexual reproduction. Although it had normal male fertility, when selfed or mated as the female in outcrosess, the Deltazif1 mutant produced small, pigmented perithecia that were sterile (lack of asci and ascospores), suggesting a female-specific role for ZIF1 during fertilization or ascus development. Similar female-specific defects in sexual reproduction were observed in the DeltaMozif1 mutant. When mated as the female, the DeltaMozif1 perithecia failed to develop long necks and asci or ascospores. The ZIF1 gene is well conserved in filamentous ascomycetes, particularly in the b-ZIP domain, which is essential for its function. Expression of ZIF1 in Magnaporthe oryzae complemented the defects of the DeltaMozif1 mutant. These results indicate that this b-ZIP transcription factor is functionally conserved in these two fungal pathogens for plant infection and sexual reproduction.

Hydrodynamic models are generally applied to describe the dynamic wetting of Newtonian or non-Newtonian fluids on a solid surface. Conversely, the molecular-kinetic paradigm is only utilized for spreading Newtonian fluids while considering the movement of a contact line as a molecular hopping process. This study extended the molecular-kinetic paradigm to the wetting behavior of non-Newtonian fluids, while assuming there are n fluid components at the contact line regime interacting simultaneously with a solid surface during front movement. The limiting cases of the derived model at slow and fast moving speeds were discussed. Moreover, the derived model was validated based on dynamic contact angle data of three carboxymethylcellulose (CMC) aqueous solutions measured using the force-balance method. Best-fit parameters were used to interpret the wetting dynamics of CMC solutions.

Predicting the burial status (the residue exposure to the lipid bilayer or buried within the protein core) of transmembrane (TM) residues of alpha-helix membrane protein (alphaHMP) is of great importance for genome-wide annotation and for experimental researchers to elucidate diverse physiological processes. In this work, we developed a new computational model that can be used for predicting the burial status of TM residues of alphaHMP. By incorporating physicochemical scales and conservation index, an efficient prediction model using least squares support vector machine (LS-SVM) was developed. The model was developed from 43 protein chains and its prediction ability was evaluated by an independent test set of other non-redundant ten protein chains. The prediction accuracy of our method was much better than the results of the reported works. Our results demonstrate that the LS-SVM prediction model incorporating structural and physicochemical features derived from sequence information could greatly improve the prediction accuracy.