Galectin‐3 expression in thyroid fine needle cytology (t‐FNAC) uncertain cases: Validation of molecular markers and technology innovation

Thyroid cancer is not very common, accounting for 1–2% of all cancers, with a population incidence of about 0.004%. Currently, the ability to discriminate between follicular adenoma and carcinoma represents the major challenge in preclinical diagnosis of thyroid proliferative lesions. Better discrimination between the two would help avoid unnecessary thyroidectomy and save valuable resources. Over the years, galectin‐3 (Gal‐3) has been proposed as a diagnostic marker with varied success. In this paper, we used Environmental Scanning Electron Microscopy Immunogold Labelling (ESEM‐IGL) to investigate the expression of Gal‐3 on Thin‐Prep fine needle aspiration cytology (FNAC). We optimized the ESEM‐IGL method on thyroid cell lines (RO‐82 and FTC‐133) comparing our membrane Gal‐3 labeling data with Western blot. We evaluated 183 thyroid FNAC from Italian patients with a uncertain pre‐surgical diagnosis. ESEM‐IGL method marker sensitivity is 71.2%, while specificity is 53.3% and diagnostic efficacy is 61.2%. Our results confirmed that Gal‐3 expression is associated with situations of hypertrophy and/or cellular hyperproliferation, pathophysiological situations common both to adenomas and to thyroid carcinomas. The innovation of thyroid FNAC Thin‐Prep ESEM‐IGL shows the levels of Gal‐3 immunolabeling clearly, even through the individual cells of a thyroid nodule. However, Gal‐3 alone, as a molecular marker of thyroid cancer, can still have a limited application in pre‐surgery diagnosis. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.     

Reperti Teratologici in Tulipa Gesneriana L. Var. Spathulata (Bert.)

Summary

Here is described by the author a teratological case of Tulipa Gesneriana L. v. spathulata (Bert.) which is notable for the concomitance of several anomalies in a single individual:

a) irregular number of the leaves;

b) scape ending with three flowers which are anomalous;

c) two from the above mentioned flowers have tetracarpellary pistil;

d) the third flower has an androecium which is formed by three external stames and, besides, by three other stames which are smaller than the first-ones and are also inglobed in a bypertrophic esacarpellary pistill.     

Affinity analysis of differentially expressed genes in hepatocytes expressing HCV core genotype 1b or 3a

Chronic hepatitis C patients display many genotype-specific clinical features of HCV infection. The core proteins encoded by different genotypes dysregulate numerous sets of distinct host genes. In this study we tested the hypothesis that HCV core proteins 1b and 3a would actually act on a limited number of independent cellular players, as well as on several functionally linked gene products. Structural and functional tests identified a core set of host genes dysregulated by HCV core genotypes 1b and 3a. The core proteins of HCV genotypes 1b and 3a target specifically limited sets of functionally related gene products, which may be responsible for the variations in the clinical spectra associated with HCV infection.     

Adjustable ‘cross-linking’ of neighboring DNA molecules in liquid-crystalline dispersions through (daunomycin-copper) polymeric chelate complexes

The interaction of daunomycin molecules with double-stranded DNA in the liquid-crystalline state was investigated. It was shown that at a certain extent of daunomycin binding a change of the mechanism of anthracycline orientation with reference to the DNA chain occurs. This is testified by the alteration of the sense of spatial packing of the DNA molecules in liquid-crystalline dispersions formed as a result of phase separation in poly(ethyleneglycol)-containing solutions, as well as by the onset of the reaction of daunomycin with divalent copper ions. Using this reaction, polymeric (daunomycin-copper) chelate cross-links between the DNA molecules fixed in the liquid-crystalline dispersions were formed. The length of such cross-links is adjusted by the distance between the DNA molecules, which, in turn, depends on the concentration of poly(ethyleneglycol) used for phase separation. The above molecular building mechanism may lead to new interesting applications.     

Fundamental Roles of the Golgi-Associated Toxoplasma Aspartyl Protease,ASP5, at the Host-Parasite Interface

Toxoplasma gondii possesses sets of dense granule proteins (GRAs) that either assemble at, or cross the parasitophorous vacuole membrane (PVM) and exhibit motifs resembling the HT/PEXEL previously identified in a repertoire of exported Plasmodium proteins. Within Plasmodium spp., cleavage of the HT/PEXEL motif by the endoplasmic reticulum-resident protease Plasmepsin V precedes trafficking to and export across the PVM of proteins involved in pathogenicity and host cell remodelling. Here, we have functionally characterized the T. gondii aspartyl protease 5 (ASP5), a Golgi-resident protease that is phylogenetically related to Plasmepsin V. We show that deletion of ASP5 causes a significant loss in parasite fitness in vitro and an altered virulence in vivo. Furthermore, we reveal that ASP5 is necessary for the cleavage of GRA16, GRA19 and GRA20 at the PEXEL-like motif. In the absence of ASP5, the intravacuolar nanotubular network disappears and several GRAs fail to localize to the PVM, while GRA16 and GRA24, both known to be targeted to the host cell nucleus, are retained within the vacuolar space. Additionally, hypermigration of dendritic cells and bradyzoite cyst wall formation are impaired, critically impacting on parasite dissemination and persistence. Overall, the absence of ASP5 dramatically compromises the parasite’s ability to modulate host signalling pathways and immune responses.     

Toxoplasma gondii exploits the host ESCRT machinery for parasite uptake of host cytosolic proteins

Toxoplasma gondii is a master manipulator capable of effectively siphoning the resources from the host cell for its intracellular subsistence. However, the molecular underpinnings of how the parasite gains resources from its host remain largely unknown. Residing within a non-fusogenic parasitophorous vacuole (PV), the parasite must acquire resources across the limiting membrane of its replicative niche, which is decorated with parasite proteins including those secreted from dense granules. We discovered a role for the host Endosomal Sorting Complex Required for Transport (ESCRT) machinery in host cytosolic protein uptake by T. gondii by disrupting host ESCRT function. We identified the transmembrane dense granule protein TgGRA14, which contains motifs homologous to the late domain motifs of HIV-1 Gag, as a candidate for the recruitment of the host ESCRT machinery to the PV membrane. Using an HIV-1 virus-like particle (VLP) release assay, we found that the motif-containing portion of TgGRA14 is sufficient to substitute for HIV-1 Gag late domain to mediate ESCRT-dependent VLP budding. We also show that TgGRA14 is proximal to and interacts with host ESCRT components and other dense granule proteins during infection. Furthermore, analysis of TgGRA14-deficient parasites revealed a marked reduction in ingestion of a host cytosolic protein compared to WT parasites. Thus, we propose a model in which T. gondii recruits the host ESCRT machinery to the PV where it can interact with TgGRA14 for the internalization of host cytosolic proteins across the PV membrane (PVM). These findings provide new insight into how T. gondii accesses contents of the host cytosol by exploiting a key pathway for vesicular budding and membrane scission.     

Linear oligopeptides. XLIII. Study of the relationship between conformation and nature of side chain: Homologous series derived from γ-branched amino acid residues

Conformational studies of the three homologous series, from dimer through heptamer, of monodisperse, N-and C-protected oligopeptides derived from the γ-branched α-amino acid residues L -leucine, β-cyclohexyl-L -alanine, and L -phenylalanine are reported. By means of ir absorption and CD in the vacuum-uv (150nm), the occurrence of intermolecular β-conformations in the higher oligomers in the solid state was established. In solvents of low polarity at high dilution, the extent of intramolecularly hydrogen-bonded folded-structure formation was assessed as a function of chain length and the nature of the side chain. Unordered and intermolecular β-conformations were found in alcohols and aqueous alcoholic mixtures. The results obtained indicate that—the position of branching being equal–steric requirements, electronic properties, and hydrophobic character of the amino acid side chains are all important in determining the nature and stability of oligopeptide conformations.