Betulaceae, Corylaceae, Cupressaceae, Fagaceae and Salicaceae around Parma (Northern Italy): Pollen calendars from 1995 to 1997

Many seasonal respiratory allergies are caused byairborne pollens. There is an evident correlationbetween allergic attacks and the amount of pollens inthe atmosphere at any time. The airbornetree pollen concentration and the relevantrepercussions on pollinotics hasnot yet been extensively reported for all Italian regions.We present the results of a 3-year, weekly, tree pollen count byBurkard spore trap from the atmosphere of Parma,Italy. Annual pollen calendars were made from theresults. The study has been conducted because, in Europe,an increasing frequency of allergic sensitisation to thesepollens has been observed.     

Preparation of a glycosynthase from the β-glycosidase of the Archaeon Pyrococcus horikoshii

The β-glycosidase from the hyperthermophilic Archaeon Pyrococcus horikoshii (Phoβ-gly) is a monomeric enzyme with wide substrate specificity belonging to family 1 of glycoside hydrolases classification. Inspection of the three-dimensional structure of the enzyme, recently resolved, showed that Phoβ-gly is membrane bound and that the residues putatively involved in the catalytic activity are Glu155 and Glu324 working as the general acid/base and the nucleophile of the reaction, respectively. We show here that mutation of the latter completely eliminated the activity of the enzyme and that it could be reactivated in the presence of sodium formate. Analysis of the products obtained in the presence of sodium formate buffer pH 4.0 at 75°C showed that the Glu324Gly mutant acts as a hyperthermophilic glycosynthase.     

The characterization of <Emphasis Type="Italic">Thermotoga maritima</Emphasis> ferritin reveals an unusual subunit dissociation behavior and efficient DNA protection from iron-mediated oxidative stress

Ferritin from the hyperthermophilic anaerobe Thermotoga maritima, a bacterium of ancient phylogenetic origin, is structurally similar to known bacterial and eukaryotic ferritins: 24 identical subunits assemble into a shell having octahedral symmetry and a Mr of about 460 kDa. T. maritima ferritin (TmFtn), purified to homogeneity as a recombinant protein, contains approximately 2–3 iron atoms and can incorporate efficiently up to 3,500 atoms in the form of a ferric oxy-hydroxide mineral at 80°C, the optimal growth temperature of the bacterium. The 24-mer unexpectedly dissociates reversibly into dimers at low ionic strengths. In turn, dimers re-associate into the native 24-mer assembly at high protein concentrations and upon incorporation of iron micelles containing at least 500 Fe(III). TmFtn uses O₂ as efficient iron oxidant. The reaction stoichiometry is 3–4 O₂:Fe(II) as in all bacterial ferritins. Accordingly no H₂O₂ is released into solution, a feature reflected in the in vitro ability of TmFtn to reduce significantly iron-mediated oxidative damage to DNA at 80°C. A similar TmFtn-mediated ROS detoxifying role likely occurs in the bacterium which lacks the SOD/catalase defense systems of the aerobic world.     

Effects of low concentrations of Regorafenib and Sorafenib on human HCC cell AFP,migration, invasion,and growth in vitro

Sorafenib was shown in clinical trial to enhance survival in hepatocellular carcinoma (HCC) patients, but with minimal tumor shrinkage. To correlate several indices of HCC growth at various drug concentrations, HCC cells were grown in various low concentrations of two multikinase inhibitors, Regorafenib (Stivarga) and Sorafenib (Nexavar) and their effects were examined on alpha‐fetoprotein (AFP), cell growth, migration, and invasion. In two AFP positive human HCC cell lines, AFP was inhibited at 0.1–1 µM drug concentrations. Cell migration and invasion were also inhibited at similar low drug concentrations. However, 10‐fold higher drug concentrations were required to inhibit cell growth in both AFP positive and negative cells. To investigate this concentration discrepancy of effects, cells were then grown for prolonged times and sub‐cultured in low drug concentrations and then their growth was re‐tested. The growth in these drug‐exposed cells was found to be slower than cells without prior drug exposure and they were also more sensitive to subsequent drug challenge. Evidence was also found for changes in cell signaling pathways in these slow‐growth cells. Low multikinase inhibitor concentrations thus modulate several aspects of HCC cell biology. J. Cell. Physiol. 228: 1344–1350, 2013. © 2012 Wiley Periodicals, Inc.     

Pelagic cephalopods of the central Mediterranean Sea determined by the analysis of the stomach content of large fish predators

The pelagic cephalopod fauna of the central Mediterranean Sea was investigated through stomach content analyses of large fish predators. A total of 124 Xiphias gladius, 22 Thunnus thynnus, 100 Thunnus alalunga, and 25 Tetrapturus belone were analyzed. Overall, 3,096 cephalopods belonging to 23 species and 16 families were identified. The cephalopod fauna in the study area is dominated by Sepiolidae, Ommastrephidae, and Onychoteuthidae. The sepiolid Heteroteuthis dispar was the most abundant species (n?=?1,402) while the ommastrephid Todarodes sagittatus showed the highest biomass. They can be considered key-species in the pelagic food web of the study area. The neutrally buoyant Histioteuthis bonnellii, H. reversa, and Chiroteuthis veranyi seem to characterize the deeper water layers. Given the difficulty in sampling pelagic cephalopods, the presence of cephalopod beaks in the stomach of predators represents a fundamental tool to assess the biodiversity and the ecological importance of these taxa in the marine ecosystem.     

Micro- and nanopatterning by lithographically controlled wetting

This protocol describes how to perform lithographically controlled wetting (LCW). LCW enables large-area patterning of microstructures and nanostructures of soluble materials, either organic or inorganic, including biological compounds in buffer solutions or compounds for cell guidance. LCW exploits the capillary forces of menisci established under the protrusions of a stamp placed in contact with a liquid film. In the space confined by each meniscus, the self-organization of the deposited solute yields highly ordered structures that replicate the motif of the stamp protrusions. The method does not require any particular infrastructure and can be accomplished by using simple tools such as compact discs or microscopy grids. Compared with other printing methods, LCW is universal for soluble materials, as it does not require chemical binding or other specific interactions between the solute and the surface. A process cycle takes from 2 to 36 h to be completed, depending on the choice of materials.     

Frequency of holoprosencephaly in the International Clearinghouse Birth Defects Surveillance Systems: searching for population variations

BACKGROUND: Holoprosencephaly (HPE) is a developmental field defect of the brain that results in incomplete separation of the cerebral hemispheres that includes less severe phenotypes, such as arhinencephaly and single median maxillary central incisor. Information on the epidemiology of HPE is limited, both because few population‐based studies have been reported, and because small studies must observe a greater number of years in order to accumulate sufficient numbers of births for a reliable estimate. METHODS: We collected data from 2000 through 2004 from 24 of the 46 Birth Defects Registry Members of the International Clearinghouse for Birth Defects Surveillance and Research. This study is based on more than 7 million births in various areas from North and South America, Europe, and Australia. RESULTS: A total of 963 HPE cases were registered, yielding an overall prevalence of 1.31 per 10,000 births. Because the estimate was heterogeneous, possible causes of variations among populations were analyzed: random variation, under‐reporting and over‐reporting bias, variation in proportion of termination of pregnancies among all registered cases and real differences among populations. CONCLUSIONS: The data do not suggest large differences in total prevalence of HPE among the studied populations that would be useful to generate etiological hypotheses. Birth Defects Research (Part A), 2008. © 2008 Wiley‐Liss, Inc.     

One drug for two targets: Biological evaluation of antiretroviral agents endowed with antiproliferative activity

AIDS-related cancer diseases are malignancies with low incidence on healthy people that affect mostly subjects already immunocompromised. The connection between HIV/AIDS and these cancers has not been established yet, but a weakened immune system is certainly the main cause. We envisaged the possibility to screen a small library of compounds synthesized in our laboratory against opportunistic tumors mainly due to HIV infection like Burkitt’s Lymphoma. From cellular assays and gene expression analysis we identified two promising compounds. These derivatives have the dual action required inhibiting HIV replication in human TZM-bl cells infected with HIV-1 NL4.3 and showing cytotoxic activity on human colon HT-29 and breast adenocarcinoma MCF-7 cells. In addition, preclinical in vitro adsorption, distribution, metabolism, and excretion studies highlighted a satisfactory pharmacokinetic profile.     

The mutation K30D disrupts the only salt bridge at the subunit interface of the homodimeric hemoglobin from Scapharca inaequivalvis and changes the mechanism of cooperativity.

The subunit interface of the homodimeric hemoglobin from Scapharca inaequivalvis, HbI, is stabilized by a network of interactions that involve several hydrogen-bonded structural water molecules, a hydrophobic patch, and a single, symmetrical salt bridge between residues Lys-30 and Asp-89. Upon mutation of Lys-30 to Asp, the interface is destabilized markedly. Sedimentation equilibrium and velocity experiments allowed the estimate of the dimerization constants for the unliganded (K(1,2D) = 8 x 10(4) M(-1)) and for the CO-bound (K(1,2L) = 1 x 10(3) m(-1)) and oxygenated (K(1,2L) = 70 m(-1)) derivatives. For the oxygenated derivative, the destabilization of the subunit interface with respect to native HbI corresponds to about 8 kcal/mol, an unexpectedly high figure. In the K30D mutant, at variance with the native protein, oxygen affinity and cooperativity are strongly dependent on protein concentration. At low protein concentrations (e.g. 1.2 x 10(-5) m heme), at which the monomeric species becomes significant also in the unliganded derivative, oxygen affinity increases and cooperativity decreases. At protein concentrations where both derivatives are dimeric (e.g. 3.3 x 10(-3) m heme), both cooperativity and oxygen affinity decrease. Taken together, the experimental data indicate that in the K30D mutant, the mechanism of cooperativity is drastically altered and is driven by a ligand-linked monomer-dimer equilibrium rather than being based on a direct heme-heme communication as in native HbI.     

Phthalazine PDE4 inhibitors. Part 3: the synthesis and in vitro evaluation of derivatives with a hydrogen bond acceptor.

This communication describes the synthesis and in vitro evaluation of a novel and potent series of phthalazine phosphodiesterase type (IV) (PDE4) inhibitors. The interaction with two distinct polar binding sites allowed us to eliminate the cyclopentyloxy substitution from rolipram-like analogues.