The protein kinase ImeB is required for light-mediated inhibition of sexual development and for mycotoxin production in Aspergillus nidulans

Spore formation is a common process in the developmental cycle of fungi. In the yeast Saccharomyces cerevisiae, Ime2 is a key protein kinase for the meiotic cell cycle, which precedes ascospore formation. Here, we analysed the IME2 -related imeB gene of the filamentous ascomycete Aspergillus nidulans. imeB deletion strains are retarded in growth and overproduce fertile sexual fruiting bodies in the presence of light, which normally represses sexual development. imeB mutants also display abnormal differentiation of sexual Hülle cells in submerged cultures. Increased sexual development of imeB mutants is dependent on VeA, a component of the heterotrimeric velvet complex. A combined deletion of imeB with the phytochrome fphA , a red light receptor, results in a complete loss of light response, suggesting that ImeB and FphA cooperate in light-mediated inhibition of sexual development. Furthermore, we found that imeB mutants fail to produce the mycotoxin sterigmatocystin, an aflatoxin precursor, and show that ImeB is needed for expression of the sterigmatocystin gene cluster. ImeB contains a TXY motif conserved in mitogen-activated protein kinases. This sequence element is essential for ImeB function. We conclude that ImeB is a mitogen-activated protein kinase-related protein kinase required for the co-ordinated control of light-dependent development with mycotoxin production.     

Chitosan film containing fucoidan as a wound dressing for dermal burn healing: Preparation and in vitro/in vivo evaluation

The aim of this study was to develop chitosan film containing fucoidan and to investigate its suitability for the treatment of dermal burns on rabbits. Porous films, thickness between 29.7 and 269.0 μm, were obtained by the solvent dropping method. Water vapor permeability (3.3–16.6/0.1 g), the swelling (0.67–1.77 g/g), tensile strength (7.1–45.8 N), and bioadhesion (0.076–1.771 mJ/cm²) of the films were determined. The thinnest films were obtained with the lowest chitosan concentration (P<.05). The water absorption capacity of the films sharply increased with the freeze-drying technique. The film having the thickness of 29.7 μm showed the highest amount of moisture permeability (16.6 g/0.1 g). Higher chitosan concentration significantly increased tensile strength of the films (P<.05). Using higher concentration of lactic acid made films more elastic and applicable, and these films were selected for in vivo studies. Seven adult male New Zealand white rabbits were used for the evaluation of the films on superficial dermal burns. Biopsy samples were taken at 7, 14, and 21 days after wounding, and each wound site was examined macroscopically and histopathologically. After 7 days treatment, fibroplasia and scar were observed on wounds treated with fucoidan-chitosan film. The best regenerated dermal papillary formation, best reepithelization, and the fastest closure of wounds were found in the fucoidan-chitosan film treatment group after 14 days compared with other treatment and control groups. It can be concluded that fucoidan-chitosan films might be a potential treatment system for dermal burns and that changing formulation variables can modulate the characterizations of the films. Published: May 24, 2007     

Polycomb Group Protein Bmi1 Is Required for Growth of RAF Driven Non-Small-Cell Lung Cancer

Background

We have previously described a RAF oncogene driven transgenic mouse model for non small cell lung cancer (NSCLC). Here we examine whether tumor initiation and growth requires the stem cell self-renewal factor Bmi1.

Principal Findings

In order to evaluate Bmi1 function in NSCLC two founder lines that differ in incidence and latency of tumor formation were compared. Ablation of Bmi1 expression in both lines had a dramatically decreased tumor growth. As the line with shorter latency matched the life span of Bmi1 knock out mice, these mice were chosen for further study. The absence of Bmi1 did not decrease the number of tumor initiation in these mice as only the size and not the number of tumors decreased. Reduction in tumor growth resulted from an increase in cell death and decrease in cell cycle progression that corresponded with up-regulation of the p16^INK4a and p19^ARF.

Significance

The data identifies Bmi1 as an important factor for expansion but not initiation of RAF driven NSCLC.     

Comparison of the Efficacy of the Two Tetracycline‐Containing Sequential Therapy Regimens for the Eradication of Helicobacter Pylori: 5 days Versus 14 days Amoxicillin

Aim: To compare the efficacy of 14‐day and 5‐day amoxicillin treatment on the eradication rate during tetracycline containing sequential H. pylori therapy, and also to compare the eradication rate of this regimen with those used in similar studies performed in Turkey. Method: This study included 112 patients infected with H. pylori that were randomized into 2 groups. In group A, patients (n = 56) received pantoprazole (40 mg BID) and amoxicillin (1 g BID) for 5 days, followed by pantoprazole (40 mg BID), tetracycline (500 mg QID), and metronidazole (500 mg TID) for the remaining 9 days. In group B, patients (n = 56) received pantoprazole (40 mg BID) and amoxicillin (1 g BID) for 5 days, followed by pantoprazole (40 mg BID), tetracycline (500 mg QID), metronidazole (500 mg TID), and amoxicillin (1 g BID) for the remaining 9 days. Eradication rates were calculated using both intention‐to‐treat (ITT) and per‐protocol (PP) analyses. Results: In all, 112 patients were subjected to ITT analysis and 109 patients completed the study. In group A, H. pylori eradication was achieved in 46 (82.1%) of the 56 patients included in the ITT analysis and in 46 (83.6%) of the 55 patients included in the PP analysis. In group B, H. pylori eradication was achieved in 44 (78.57%) of the 56 patients included in the ITT analysis and in 44 (81.48%) of the 54 patients included in the PP analysis ( Table 2 ). The eradication rates were not statistically significant between the 2 groups (p > .005).

Table 2. Eradication rates in the two study groups

Microfluidic immobilization of physiologically active Caenorhabditis elegans

We present a protocol for building and operating a microfluidic device for mechanical immobilization of Caenorhabditis elegans in its physiologically active state. The system can be used for in vivo imaging of dynamic cellular processes such as cell division and migration, degeneration, aging and regeneration, as well as for laser microsurgery, Ca2+ imaging and three-dimensional microscopy. The device linearly orients C. elegans, and then completely restrains its motion by pressing a flexible membrane against the animal. This technique does not involve any potentially harmful anesthetics, gases or cooling procedures. The system can be installed on any microscope and operated using only one syringe and one external valve, making it accessible to most laboratories. The device fabrication begins by patterning photoresist structures on silicon wafers, which are then used to mold features in elastomeric layers that are thermally bonded to form the device. The system can be assembled within 3 d.     

Triptolide inhibits CD133+/CD44+ colon cancer stem cell growth and migration through triggering apoptosis and represses epithelial-mesenchymal transition via downregulating expressions of snail,slug, and twist

High recurrence and metastatic behavior patterns are the most important reasons for the failure of treatment strategies in patients with colon cancer. Cancer stem cells (CSCs), which are considered root of cancer, are thought to be associated with therapy resistance, relapse, and metastasis, and, therefore, targeting CSCs rather than the bulk population may be an effective approach. In cancer studies, there is an increasing interest in close friendship between epithelial-mesenchymal transition (EMT) and CSCs. Triptolide (TPL) isolated from Chinese herb Tripterygium wilfordii has important effects on the prevention of migration and metastasis as well as cytotoxic effect against cancer cells. The potential lethal efficacy of TPL on CSCs that is highly resistant to the drug is an unsolved mystery. Fundamentally, the present study basically aims to find answers to two questions: (a) is it possible to target colon CSCs with TPL? and (b) what are the mechanisms underlying TPL's potential to eliminate CSCs? Cytotoxic effects of TPL on CSCs were evaluated by WST-1 and Muse count and viability assays. Apoptosis assay and cell-cycle analysis were performed to investigate the inhibitory effect of TPL. Moreover, the effects of TPL on spheroid formation capacity, migration, and EMT processes, which are associated with CSC phenotype, were also investigated. The results revealed that TPL triggered cell death and apoptosis and altered cell cycle distribution. Moreover, TPL significantly reduced the snail slug and twist expressions associated with EMT. TPL has been shown to be effective in colon CSCs by in vitro experiments, and it might be a highly effective agent against colon cancer has been implicated in need of supporting in vivo and clinical studies.     

Natal origin of Namibian grey whale implies new distance record for in-water migration

We use genomics to identify the natal origin of a grey whale found in the South Atlantic, at least 20 000 km from the species core range (halfway around the world). The data indicate an origin in the North Pacific, possibly from the endangered western North Pacific population, thought to include only approximately 200 individuals. This contributes to our understanding of Atlantic sightings of this species known primarily from the North Pacific, and could have conservation implications if grey whales have the potential for essentially global dispersion. More broadly, documenting and understanding rare extreme migration events have potential implications for the understanding of how a species may be able to respond to global change.     

Effects of a brain tumor in a dispersive human head on SAR and temperature rise distributions due to RF sources at 4G and 5G frequencies

In this paper, effects of a brain tumor located in a dispersive human head model on specific absorption rate (SAR) and temperature rise distributions due to different types of RF sources at 4G and 5G cellular frequencies are investigated with the use of a multiphysics model. This multiphysics model analyzes the dispersive human head with the brain tumor and provides the SAR and temperature rise distributions in the head due to the RF source operated at 4G and 5G cellular frequencies in a single finite-difference time-domain simulation. An adjacent antenna operated at 4G and 5G cellular frequencies to the human head is considered as the RF source for near-field exposure, while a plane wave field radiated by base stations operated at 4G and 5G cellular frequencies is considered as the RF source for far-field exposure. Numerical results show that the brain tumor in the head slightly affects the SAR and temperature rise distributions due to different RF sources at 4G and 5G cellular frequencies.     

Differential expression of proliferative, cytoskeletal, and adhesive proteins during postnatal development of the hamster submandibular gland

 Although the submandibular gland (SMG) plays important exocrine and endocrine roles, little is known about the molecular details underlying its development. Previously, we reported that in the postnatally developing hamster SMG, GPT, the protein product of the first N-glycosylation gene, ALG7, was an in vivo marker for salivary cell proliferation. Here we investigated the proliferative, cytoskeletal, and adhesive changes during SMG postnatal development. The cellular localization and abundance of GPT, filamentous actin, and β1 integrin receptor were examined using confocal microscopy and immunoblotting. In neonatal glands, high GPT levels marked extensive cell proliferation throughout the tissue. The apical regions of immature salivary cells displayed intense actin staining, while most of the β1 integrin was diffusely distributed throughout the tissue. As development proceeded, discrete regions of the gland expressed attenuated levels of GPT, an increased organization of actin to the cell cortex, and β1 integrin to the basal lamina. In the adult SMG, differentiated salivary cells displayed low levels of GPT and actin. While the abundance of β1 integrin remained unchanged throughout development, in the adult, it was found exclusively in regions where cells contact the basal lamina. These data indicate that SMG development entails regionalized cell proliferation and polarization, and that these processes are temporally and spatially coordinated with the establishment of stable cell-substratum interactions. Accepted: 26 October 1998