Biomimetic organization: Octapeptide self-assembly into nanotubes of viral capsid-like dimension

The controlled self-assembly of complex molecules into well defined hierarchical structures is a promising route for fabricating nanostructures. These nanoscale structures can be realized by naturally occurring proteins such as tobacco mosaic virus, capsid proteins, tubulin, actin, etc. Here, we report a simple alternative method based on self-assembling nanotubes formed by a synthetic therapeutic octapeptide, Lanreotide in water. We used a multidisciplinary approach involving optical and electron microscopies, vibrational spectroscopies, and small and wide angle x-ray scattering to elucidate the hierarchy of structures exhibited by this system. The results revealed the hexagonal packing of nanotubes, and high degree of monodispersity in the tube diameter (244 A) and wall thickness (approximately equal to 18 A). Moreover, the diameter is tunable by suitable modifications in the molecular structure. The self-assembly of the nanotubes occurs through the association of beta-sheets driven by amphiphilicity and a systematic aromatic/aliphatic side chain segregation. This original and simple system is a unique example for the study of complex self-assembling processes generated by de novo molecules or amyloid peptides.     

The effect of adrenaline,tyramine and guanethidine on two-way avoidance conditioning and on pseudoconditioning

Adrenaline (0.5 mg/kg) or guanethidine (10 mg/kg) given i.p. depressed performance of pseudoconditioned shuttle responses by rats. In a previous paper it had been shown that tyramine (5 mg/kg) had an opposite effect (Izquierdo, 1974a). Pre-trial administration of any of the three drugs also depressed two-way avoidance conditioning. Following posttrial administration, only guanethidine had a deleterious effect on reténtion. Since none of these drugs is believed to reach the brain in significant amounts following systemic injection, the present results suggest that peripheral factors may influence both conditioned and pseudoconditioned shuttle behavior.     

Ein Fall tätowirter menschlicher Hornhaut histologisch untersucht

Ohne Zusammenfassung     

Mucin production determines sensitivity to bortezomib and gemcitabine in pancreatic cancer cells

The prognosis of pancreatic cancer remains disappointing due to a high intrinsic resistance against chemotherapeutic agents. Standard gemcitabine therapies have improved overall survival only marginally and recently, inhibition of the proteasome by the boronic acid derivative bortezomib has been introduced as a novel therapeutic strategy for solid and hematological malignancies including pancreatic cancer. The mucus-producing pancreatic cancer cell line Capan-1 was cultured under standard conditions and treated with different concentrations of gemcitabine or bortezomib. Mucus production was suppressed by siRNA-mediated silencing of apomucin genes. Cell proliferation was determined by 3H-thymidine incorporation and apoptosis was quantified after propidium iodide staining by flow cytometry. Apoptotic cell death was confirmed by TUNEL staining, determination of mitochondrial transmembrane potential and assessment of caspase 3/7 activity. NFκB-activity was determined by EMSA. The unfolded protein response (UPR) was further investigated by PCR, Western blotting and caspase 12 activity assays. Silencing of MUC4 significantly reduced expression of mucins for up to 5 days after transfection. While native cells showed an increased sensitivity to bortezomib treatment, silenced cells were more sensitive to gemcitabine treatment. Bortezomib induces mitochondrial damage in native cells and also activates the UPR by splicing of Xbp-1 and induction of CHOP, which is significantly reduced by silencing of MUC4. Our data suggest that mucinous pancreatic cancers are more sensitive towards proteasome inhibition by bortezomib and that alternative pathways of apoptosis are involved in cell death induction, while tumor cells with a low secretory activity show a better response to gemcitabine.     

Uptake,accumulation and release of ouabain by isolated rat hepatocytes

Summary We investigated uptake of ouabain into isolated rat hepatocytes and release of ouabain from preloaded hepatocytes, thus assessing separately the two membrane transport steps, involved in biliary elimination of the drug. The following results were obtained:Uptake of ouabain was saturable (K _m=263±61M, V=3.3±1.0 nmol/mg prot.×min), energy-dependent, sensitive to dinitrofluorobenzene and temperature-dependent (E=80–96 kJ/mol). It had no pH-optimum in the physiological pH-range and was independent of the extracellular cation composition.Uptake of ouabain was competitively inhibited by the cardiac glycoside digitoxin (K _i=0.3 M).Uptake was not inhibited in the presence of the glycosidic sugar l-rhamnose, but it was competitively inhibited by the steroid taurocholate (K _i=6.3 M).Ouabain was accumulated within hepatocytes 170-fold. The predominant fraction of intracellular ouabain beeing unbound.Release of ouabain from preloaded cells was energyindependent, independent of the Na⁺-concentration and not susceptible to inhibition by dinitrofluorobenzene or taurocholate.It is concluded, first that hepatocellular uptake of ouabain is mediated by a carrier for steroids and second that the pathway of release is distinct from that of uptake. We assume, that the high bile/plasma concentration-gradient of ouabain in vivo is generated during active uptake into the cell and not during release into bile.     

Substituted 2-aminothiophenes: antifungal activities and effect on Microsporum gypseum protein profile

The increasing recognition and importance of fungal infections, the difficulties encountered in their treatment and the increase in resistance to antifungal agents have stimulated the search for therapeutic alternatives. The objective of this study was to evaluate the antifungal activities of three substituted 2‐aminothiophenes (1, 2 and 3) against some fungal species. The synthesis of substituted 2‐aminothiophenes was carried out through the most versatile synthetic method developed by Gewald et al. The antifungal activity was performed against yeast, dermatophytes and Aspergillus species using the broth microdilution method. The effect of these aminothiophenes was examined on the protein content and profile. Compound 2 was the most active (MIC varying from 2.00 to 128 μg ml^?1). All the three substituted 2‐aminothiophenes had a relatively important dose‐dependent effect on Microsporum gypseum protein profile and content. These compounds affected the structure and dye fixation of macroconidia of this fungus. The overall results indicate that the tested substituted 2‐aminothiophenes can be used as precursors for new antifungal drugs development.     

A model for the function of the bisphosphorylated heart-specific troponin-I N-terminus

Bisphosphorylation of two adjacently located serine residues in the heart-specific N-terminus of the cTnI subunit reduces calcium affinity of the cTnC subunit. An interaction of the phosphorylation region of cTnI with acidic residues of another cTn subunit has been proposed formerly based on 31P nuclear magnetic resonance (NMR) data. A possible candidate is cTnC. Thus, an interaction model of cTnC with the bisphosphorylated cTnI N-terminus has been built using a homology model of hcTnC based on the crystal structure of tusTnC and the structure of the phosphorylation region of cTnI determined by 2D NMR. By computational search, five clusters of acidic residues on cTnC might interact with the cTnI phosphorylation region. Three sites could be excluded by 31P-NMR experiments. The two remaining sites are located in the N-terminal helix of cTnC and between calcium binding sites III and IV. Reorientation of the arginine and phosphoserine sidechains within thephosphorylation region as proposed by refined docking could explain the formerly measured changes in pKa app values. Thus, local pKa changes might lead to the reduction of calcium affinity observed upon cTnI bisphosphorylation.This revised version was published online in September 2005 with corrections to the Cover Date.     

Behavior Change,Environmental Hazards and Respiratory Protection Among a Southern Farm Community

ABSTRACT

The agricultural industry ranks as one of the most dangerous in terms of occupational deaths and injuries. A wide variety of respiratory illnesses can result from the exposure to grain and organic dusts and working in animal confinement facilities and barns. This article analyzes the Transtheoretical Model of Change for implications relevant to health promotion and education. This study explored differences of perception of occupational health, environmental exposures, and stages of change consistent with the readiness to take action and the confidence to act toward respiratory health among farmers who report respiratory symptoms with physical activity and those who do not. A convenience sample of 123 farm owners and agricultural employees recruited from community-based agricultural events in southeast and central Louisiana completed three surveys: (a) Health Risk and Environmental Assessment; (b) Identification of Respiratory Mask Use; (c) Stage of Awareness and Preventive Respiratory Health; and demographic information. Subjects performed pulmonary function tests including three Forced Vital Capacity (FVC) maneuvers while connected to a Renaissance spirometer. Even though the majority of subjects rated themselves in action stage of change, subjects reported using respiratory protective devices less than 10% of the time. No significant differences were found in environmental exposures. Using chi-square analysis, those farmers who reported breathlessness were significantly more likely to report fatigue, chest pain, and dizziness. Those farmers who report breathlessness are significantly less likely to perceive respiratory health as important compared to other occupational illnesses/conditions. There is a need for additional studies to further examine the relationship between respiratory symptoms, exposure risks, and behavior change theory.     

A single-arm phase II Austrian/German multicenter trial on continuous daily sunitinib in primary glioblastoma at first recurrence (SURGE 01-07)

Background

Due to the redundancy of molecular pathways simultaneously involved in glioblastoma growth and angiogenesis, therapeutic approaches intervening at multiple levels seem particularly appealing.

Methods

This prospective, multicenter, single-arm phase II trial was designed to evaluate the antitumor activity of sunitinib, an oral small-molecule inhibitor of several receptor tyrosine kinases, in patients with first recurrence of primary glioblastoma using a continuous once-daily dosing regimen. Patients received a starting dose of sunitinib 37.5 mg, followed by a maintenance dose between 12.5 mg and 50 mg depending on drug tolerability. The primary endpoint was a 6-month progression-free survival (PFS) rate. Secondary endpoints included median PFS, overall survival (OS), safety/toxicity, quality of life, and translational studies on the expression of sunitinib target molecules.

Results

Forty participants were included in this study, and no objective responses were detected. PFS6 was 12.5%, median PFS 2.2 months, and median OS 9.2 months. Five participants (12.5%) showed prolonged stable disease ≥6 months with a median PFS of 16.0 months (range, 6.4–41.4 mo) and a median OS of 46.9 months (range, 21.2–49.2 mo) for this subgroup. c-KIT expression in vascular endothelial cells (n = 14 participants) was associated with improved PFS. The most common toxicities were fatigue/asthenia, mucositis/dermatitis, dysesthesias, gastrointestinal symptoms, cognitive impairment, leukoctopenia, and thrombocytopenia. Two participants (5%) terminated treatment due to toxicity.

Conclusion

Continuous daily sunitinib showed minimal antiglioblastoma activity and substantial toxicity when given at higher doses. High endothelial c-KIT expression may define a subgroup of patients who will benefit from sunitinib treatment by achieving prolonged PFS.ClinicalTrials.gov Identifier: {"type":"clinical-trial","attrs":{"text":"NCT00535379","term_id":"NCT00535379"}}NCT00535379.