Targeted immunomodulation of the NF-kappaB pathway in airway epithelium impacts host defense against Pseudomonas aeruginosa

We investigated the impact of inflammatory signaling in airway epithelial cells on host defense against Pseudomonas aeruginosa, a major cause of nosocomial pneumonia. In mice, airway instillation of P. aeruginosa resulted in NF-kappaB activation in the lungs that was primarily localized to the bronchial epithelium at 4 h, but was present in a variety of cell types by 24 h. We modulated NF-kappaB activity in airway epithelium by intratracheal delivery of adenoviral vectors expressing RelA (AdRelA) or a dominant inhibitor of NF-kappaB before P. aeruginosa infection. Bacterial clearance was enhanced by up-regulation of NF-kappaB activity following AdRelA administration and was impaired by treatment with a dominant inhibitor of NF-kappaB. The TNF-alpha concentration in lung lavage was increased by AdRelA treatment and beneficial effects of NF-kappaB up-regulation were abrogated in TNF-alpha-deficient mice. In contrast, NF-kappaB inhibition reduced MIP-2 expression and neutrophil influx following P. aeruginosa infection. Therefore, inflammatory signaling through the NF-kappaB pathway in airway epithelial cells critically regulates the innate immune response to P. aeruginosa.     

Comparative proteomic approach to identify proteins involved in flooding combined with salinity stress in soybean

Salinity together with waterlogging or flooding, a condition that occurs frequently in the field, can cause severe damage to crops. Combined flooding and salinity decreases the growth and survival of plants more than either stress alone. We report here the first proteomic analysis to investigate the global effects of saline flooding on multiple metabolic pathways. Soybean seedlings at the emergence (VE) stage were treated with 100 mM NaCl and flooded with water or 100 mM sodium chloride solution for 2 days. Proteins were extracted from hypocotyl and root samples and analyzed by two-dimensional gel electrophoresis followed by MALDI-TOF, MALDI-TOF/TOF mass spectrometry or immunoblotting. A total of 43 reproducibly resolved, differentially expressed protein spots visualized by Coomassie brilliant blue staining were identified by MALDI-TOF MS. Identities of several proteins were also validated by MS/MS analysis or immunoblot analysis. Twenty-nine proteins were upregulated, eight proteins were downregulated and six spots were newly induced. The identified proteins include well-known salt and flooding induced proteins as well as novel proteins expressed by the salinity-flooding combined stress. The comparative analysis identified changes at the proteome level that are both specific and part of a common or shared response. The identification of such differentially expressed proteins provides new targets for future studies that will allow assessment of their physiological roles and significance in the response of glycophytes to a combination of flooding and salinity.     

Acoustic Characteristics of Stridor in Multiple System Atrophy

Nocturnal stridor is a breathing disorder prevalent in patients with multiple system atrophy (MSA). An improved understanding of this breathing disorder is essential since nocturnal stridor carries a poor prognosis (an increased risk of sudden death). In this study, we aimed to classify types of stridor by sound analysis and to reveal their clinical significance. Patients who met the criteria for probable MSA and had undergone polysomnography (PSG) were recruited. Patients were then assessed clinically with sleep questionnaires, including the Pittsburgh Sleep Quality Index, and the Hoehn and Yahr scale. Nocturnal stridor and snoring were analyzed with the Multi-Dimensional Voice Program. Nocturnal stridor was recorded in 22 patients and snoring in 18 patients using the PSG. Waveforms of stridors were classified into rhythmic or semirhythmic after analysis of the oscillogram. Formants and harmonics were observed in both types of stridor, but not in snoring. Of the 22 patients diagnosed with stridor during the present study, fifteen have subsequently died, with the time to death after the PSG study being 1.9 ± 1.4 years (range 0.8 to 5.0 years). The rhythmic waveform group presented higher scores on the Hoehn and Yahr scale and the survival outcome of this group was lower compared to the semirhythmic waveform group (p = 0.030, p = 0.014). In the Kaplan Meier’s survival curve, the outcome of patients with rhythmic waveform was significantly less favorable than the outcome of patients with semirhythmic waveform (log-rank test, p < 0.001). Stridor in MSA can be classified into rhythmic and semirhythmic types and the rhythmic component signifies a poorer outcome.     

Automatic protein structure prediction system enabling rapid and accurate model building for enzyme screening

Protein structure prediction has great potential of understanding the function of proteins at the molecular level and designing novel protein functions. Here, we report rapid and accurate structure prediction system running in an automated manner. Since fold recognition of the target protein to be modeled is the starting point of the template-guided model building process, various approaches – such as profile analysis, threading, and SCOP fold classification – have been applied to generate the template library and to select the best template structure. After the best template was determined, fold consistency within the template candidates was considered using TM-score and SCOP database to select additional template structures among the template library. To generate a total of 100 decoy sets, MODELLER was used with the selected template structure. The predicted decoys were clustered with the RMSD deviation criterion of 3 Å to obtain centroids from each cluster. Finally, the selected centroids were subject to side-chain rearrangement using SCWRL module. Our fully automated structure prediction system was examined with sample test sets consisting of recently released 80 PDB chains. Judged by the TM-score (≥0.4), we concluded that 60 cases (75%) showed similar structures of statistical significance. This prediction system provides the users with simple and reliable models within hours of query submission, so that it is quite simply used for high throughput enzyme screening.     

Hair Iron and Other Minerals’ Level in Breast Cancer Patients

Little is known about hair minerals in cancer patients, and serum iron level has been shown to be elevated in breast cancer patients. Therefore, the aim of this study was to evaluate hair iron and hair minerals’ level related to hair iron in breast cancer patients compared to controls. We compared hair mineral analysis data of 40 breast cancer subjects with age and body mass index-matched normal control data (n?=?144) by cross-sectional analysis. All breast cancer patients were newly diagnosed at one Breast Cancer Center in Ajou University and had their hair cut before anti-cancer chemotherapy, and the normal controls (without breast cancer) also had their hair cut for various reasons in out-patient clinics of the Department of Family Practice and Community Health. Breast cancer patients had low calcium, magnesium, iron, copper, manganese, and zinc, whereas they had high arsenic, sodium, and potassium compared with the normal control. The hair iron level was positively correlated with hair calcium (r?=?0.761, P?<?0.001), magnesium (r?=?0.643, P?<?0.001), and manganese (r?=?0.550, P?<?0.001) and negatively correlated with arsenic (r?=??0.537, P?<?0.001). The hair iron level was significantly associated with the hair calcium (β?=?0.778, P?<?0.001) and manganese (β?=?0.240, P?<?0.001) by using multiple linear regression analysis. We observed different hair mineral patterns in breast cancer patients compared to normal controls. Especially, hair iron level was significantly reduced and associated with hair calcium and manganese levels.     

Purification and characterization of an extracellular polysaccharide from haloalkalophilic Bacillus sp. I-450

During the course of investigation of haloalkalophilic bacteria, we screened some heavily polluted soil samples from the mudflats surrounding the city of Inchon, Korea, for their bioflocculant producing ability. Based on the screening, one isolate no. 450 tentatively identified as Bacillus sp. produced an extracellular polysaccharide having flocculation activity. The isolate produced the polysaccharide during the late logarithmic growth phase. The polymer could be recovered from the supernatant of the fermented medium by cold ethanol precipitation and purified by treating with cetylpyridinium chloride (CPC). The polymer was identified as an acidic polysaccharide containing neutral sugars, namely, galactose, fructose, glucose and raffinose, and uronic acids as major and minor components, respectively. The amount of neutral sugars, uronic acid and amino sugars were 52.4, 17.2 and 2.4%, respectively. The molecular weight of the polysaccharide was found to be 2.2×10⁶ Da. The Fourier transform infrared spectrophotometer (FT-IR) revealed typical characteristics of polysaccharides. ¹H NMR spectrum showed that the polymer is a heteroglycan. Thermogravimetric (TGA) analysis indicated the degradation temperature (T_d) at 290 °C. The rheological analysis of the polymer 450 revealed the pseudoplastic property with shear-thinning effect, while the compression test indicated that the polymer had high gel strength, and the S.E.M. studies showed that the polymer has a porous structure with small pore-size distribution indicating the compactness of the polymer.     

Anticancer activity of TTAC-0001, a fully human anti-vascular endothelial growth factor receptor 2 (VEGFR-2/KDR) monoclonal antibody,is associated with inhibition of tumor angiogenesis

Vascular endothelial growth factor (VEGF) and its receptors are considered the primary cause of tumor-induced angiogenesis. Specifically, VEGFR-2/kinase insert domain receptor (KDR) is part of the major signaling pathway that plays a significant role in tumor angiogenesis, which is associated with the development of various types of tumor and metastasis. In particular, KDR is involved in tumor angiogenesis as well as cancer cell growth and survival. In this study, we evaluated the therapeutic potential of TTAC-0001, a fully human antibody against VEGFR-2/KDR. To assess the efficacy of the antibody and pharmacokinetic (PK) relationship in vivo, we tested the potency of TTAC-0001 in glioblastoma and colorectal cancer xenograft models. Antitumor activity of TTAC-0001 in preclinical models correlated with tumor growth arrest, induction of tumor cell apoptosis, and inhibition of angiogenesis. We also evaluated the combination effect of TTAC-0001 with a chemotherapeutic agent in xenograft models. We were able to determine the relationship between PK and the efficacy of TTAC-0001 through in vivo single-dose PK study. Taken together, our data suggest that targeting VEGFR-2 with TTAC-0001 could be a promising approach for cancer treatment.