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Il contributo sottolinea che il rapporto tra storici, storici dell’arte e fonti iconografiche e piu risalente rispetto al cosiddetto pictorial turn e alla recente configurazione dei visual studies. Con riferimento agli studi sul lungo ottocento addita poi come compito degli storici la ricostruzione di pratiche percettive ed esperienze concrete di fruizione delle immagini e lo studio dei margini di autonomia della dimensione visuale.

The molecular properties and subcellular location of bound gamma-glutamyl transferase (GGT) were studied, and an experimental setup devised to assess its functions in barley roots. Enzyme histochemistry was used to detect GGT activity at tissue level; immunocytochemistry to localize the protein at subcellular level; and modelling studies to investigate its surface charge properties. GGT activity in vivo was measured for the first time. Functions were explored by applying chemical treatments with inhibitors and the thiol-oxidizing drug diamide, performing time-course chromatographic and spectrophotometric analyses on low-molecular-weight thiols. Gamma-glutamyl transferase activity was found to be high in the root apical region and the protein was anchored to root cell wall components, probably by basic amino acid residues. The results show that GGT is essential to the recovery of apoplastic glutathione provided exogenously or extruded by oxidative treatment. It is demonstrated that GGT activity helps to salvage extracellular glutathione and may contribute to redox control of the extracellular environment, thus providing evidence of a functional role for gamma-glutamyl cycle in roots.

Alternative bearing surfaces for total hip arthroplasty, such as metal-on-metal and ceramic-on-ceramic, offer the potential to reduce mechanical wear and osteolysis. In the short and medium term, the second generation of metal-on-metal bearings demonstrated high systemic metal ion levels, whereas ceramic-on-ceramic bearings showed the lowest ones. We aimed to verify whether the long-term ion release in metal-on-metal subjects was still relevant at a median 10-year follow-up, and whether a fretting process at the modular junctions occurred in ceramic-on-ceramic patients and induced an ion dissemination. Serum levels were measured in 32 patients with alumina-on-alumina implants (group A), in 16 subjects with metal-on-metal implants (group B), and in 47 healthy subjects (group C). Group B results were compared with medium-term findings. Cobalt and chromium levels were significantly higher in metal-on-metal implants than in ceramic-on-ceramic ones and controls. Nevertheless, ion levels showed a tendency to decrease in comparison with medium-term content. In ceramic-on-ceramic implants, ion values were not significantly different from controls. Both in groups A and B, aluminum and titanium release were not significantly different from controls. In conclusion, negligible serum metal ion content was revealed in ceramic-on-ceramic patients. On the contrary, due to the higher ion release, metal-on-metal coupling must be prudently considered, especially in young patients, in order to obtain definitive conclusions.

Humic substances are known to affect plant metabolism at different levels. We characterized humic substances extracted from earthworm feces by diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy and used them to treat corn, Zea mays L., seedlings to investigate changes in patterns of root protein expression. After root plasma membrane extraction and purification, proteins were separated by two-dimensional gel electrophoresis, and differential spot intensities were evaluated by image analysis. Finally, 42 differentially expressed proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The majority of them were downregulated by the treatment with humic substances. The proteins identified included malate dehydrogenase, ATPases, cytoskeleton proteins, and different enzymes belonging to the glycolytic/gluconeogenic pathways and sucrose metabolism. The identification of factors involved in plant responses to humic substances may improve our understanding of plant-soil cross-talk, and enable a better management of soil resources.

OBJECTIVE—Perforin plays a key role in cell-mediated cytotoxicity. Mutations of its gene, PRF1, cause familial hemophagocytic lymphohistiocytosis but have also been associated with lymphomas and the autoimmune/lymphoproliferative syndrome. The aim of this work was to investigate the role of PRF1 variations in type 1 diabetes. RESEARCH DESIGN AND METHODS—We typed for the N252S and A91V variations in an initial population of 352 type 1 diabetic patients and 816 control subjects and a second population of 365 patients and 964 control subjects. Moreover, we sequenced the coding sequence and intron-exons boundaries in 200 patients and 300 control subjects. RESULTS—In both cohorts, allelic frequency of N252S was significantly higher in patients than in control subjects (combined cohorts: 1.5 vs. 0.4%; odds ratio 6.68 [95% CI 1.83–7.48]). Sequencing of the entire coding region detected one novel mutation in one patient, causing a P477A amino acid change not detected in 199 patients and 300 control subjects. Typing for HLA-DQA1 and DQB1 alleles showed that type 1 diabetes–predisposing DQα/DQβ heterodimers were less frequent in patients carrying N252S or P477A than in those carrying wild-type PRF1. We previously found that natural killer (NK) activity is not decreased in most N252S heterozygotes, but we detected one whose NK activity was normal at the age of 12 but strikingly low in early childhood. Here, we discovered that NK function was low in three heterozygotes in early childhood, one homozygous adult, and in the subject carrying P477A. CONCLUSIONS—These data suggest that N252S and possibly other PRF1 variations are susceptibility factors for type 1 diabetes development.

We report the proteomic characterization of the venoms of two closely related pit vipers of the genus Lachesis, L. muta (South American Bushmaster) and L. stenophrys (Central American Bushmaster), and compare the toxin repertoire of the former revealed through a proteomic versus a transcriptomic approach. The protein composition of the venoms of Lachesis muta and L. stenophrys were analyzed by RP-HPLC, N-terminal sequencing, MALDI-TOF peptide mass fingerprinting and CID-MS/MS. Around 30–40 proteins of molecular masses in the range of 13–110 kDa and belonging, respectively, to only 8 and 7 toxin families were identified in L. muta and L. stenophrys venoms. In addition, both venoms contained a large number of bradykinin-potentiating peptides (BPP) and a C-type natriuretic peptide (C-NP). BPPs and C-NP comprised around 15% of the total venom proteins. In both species, the most abundant proteins were Zn2+-metalloproteinases (32–38%) and serine proteinases (25–31%), followed by PLA2s (9–12%), galactose-specific C-type lectin (4–8%), l-amino acid oxidase (LAO, 3–5%), CRISP (1.8%; found in L. muta but not in L. stenophrys), and NGF (0.6%). On the other hand, only six L. muta venom-secreted proteins matched any of the previously reported 11 partial or full-length venom gland transcripts, and venom proteome and transcriptome depart in their relative abundances of different toxin families. As expected from their close phylogenetic relationship, the venoms of L. muta and L. stenophrys share (or contain highly similar) proteins, in particular BPPs, serine proteinases, a galactose-specific C-type lectin, and LAO. However, they dramatically depart in their respective PLA2 complement. Intraspecific quantitative and qualitative differences in the expression of PLA2 molecules were found when the venoms of five L. muta specimens (3 from Bolivia and 2 from Peru) and the venom of the same species purchased from Sigma were compared. These observations indicate that these class of toxins represents a rapidly-evolving gene family, and suggests that functional differences due to structural changes in PLA2s molecules among these snakes may have been a hallmark during speciation and adaptation of diverging snake populations to new ecological niches, or competition for resources in existing ones. Our data may contribute to a deeper understanding of the biology and ecology of these snakes, and may also serve as a starting point for studying structure–function correlations of individual toxins. Ministerio de Educación y Ciencia/[BFU2004-01432/BMC]//España Consejo Superior de Investigaciones Científicas/[CSIC-UCR 2006CR0010]/CSIC-UCR/España UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto Clodomiro Picado (ICP)

The Fas death receptor triggers lymphocyte apoptosis through an extrinsic and an intrinsic pathway involving caspase-8 and -9 respectively. Inherited defects of Fas function are displayed by a proportion of patients with Type 1 diabetes mellitus (T1DM) especially those with a second autoimmunity (T1DM-p). This study assesses activation of both pathways in Fas-resistant (FasR) patients to localize the defect. 21/28 (75 percent) T1DM-p, 14/50 (38 percent) T1DM, and 7/150 (5 percent) controls were FasR. Analysis of the 35 FasR patients and 20 Fas-sensitive (FasS) controls showed that caspase-9 activity was lower in T1DM-p and T1DM than in controls, whereas caspase-8 activity was lower in T1DM-p than in T1DM and the controls. Single patient analysis showed that 16/35 patients displayed defective activity of one (FasR1), whereas 19 displayed normal activity of both caspases (FasR2). Ages at onset of diabetes mellitus in T1DM and the second autoimmune disease in T1DM-p were lower in FasR than in FasS patients. All FasR1 patients developed diabetes mellitus before the age of 9 years, whereas a later onset was displayed by 26% FasR2 and 53% FasS patients. These data show that defective Fas function may involve both the extrinsic and intrinsic pathway in T1DM and severity correlates with the precocity of the autoimmune attack and its tissue polyreactivity.

Background Several guidelines on adeno-tonsillar disease have been proposed in recent years and some discrepancies in relation both to clinical manifestations and indications for surgical treatment have emerged. The aim of the study was to verify what influence (adeno)-tonsillectomy guidelines have had on the clinical behaviour of ENT specialists in Italy. Our study is a retrospective and multi-centre case series with chart review. Methods The survey involved 14,770 children, aged between the ages of 2 and 11, who had undergone adeno-tonsillar surgery between 2002 and 2008 in fourteen Italian tertiary and secondary referral centres. Anova test was used for the statistical analysis, assuming p < 0.05 as the minimum statistical significance value. Results The frequency of adeno-tonsillar surgeries did not change significantly (p>0.05) during the study period and following the Italian policy document publication. Overall, adeno-tonsillectomy was the most frequent intervention (64.1%), followed by adenoidectomy (31.1%) and tonsillectomy (4.8%). The indications for surgery did not change significantly for each of the operations (p>0.05), with the exception of adeno-tonsillectomy in case of feverish episodes due to acute recurrent tonsillitis ≥ 5 without nasal obstruction (decreased p= 0.010) , even when the feverish episodes due to acute recurrent tonsillitis were < 5 over the last year. Nasal obstruction was associated with feverish episodes due to acute recurrent tonsillitis in 65.2% of operated cases, while otitis media had been diagnosed in 43.3% of the patients studied. Conclusions The recommendations first developed in Italy in a 2003 policy document and then resumed in guidelines in 2008, were not implemented by ENT units involved in the survey. The study highlights the fact that the indications for adeno-tonsillar operations are based on the overall clinical presentation (comorbidity) rather than on a single symptom. Guidelines are necessary to give coherent recommendations based on both the findings obtained through randomized controlled trials and the data collected from observational studies.

12 páginas, 7 figuras -- PAGS nros. 2601-2612, Several sets of data indicate that ICOS regulates cytokine production in activated T cells, but is less effective on naïve T cells. This work evaluates ICOS function in human naïve CD4+ T cells through an assessment of the effect of soluble forms of the ICOS and CD28 physiological ligands on activation driven by anti-CD3 mAb. ICOS strikingly potentiated secretion of IL-2, IFN-γ, IL-10, and TNF-α, but not IL-4, promoted by optimal stimulation of CD3+CD28, and it was the key switching-factor of activation when cells received suboptimal stimulation of CD3+CD28 or stimulation of CD3 alone in the presence of exogenous IL-2. In these conditions, blockade of IL-2 and IFN-γ showed that ICOS builds up a positive feedback loop with IFN-γ, which required IL-2 and was inhibited by IL-4. By contrast, in the absence of CD28 triggering or exogenous IL-2, ICOS-induced costimulation mainly supported expression of TGF-β1 and FoxP3 and differentiation of regulatory T cells capable to inhibit proliferation of naïve CD4+ T cells driven by allogeneic cells. These data suggest that ICOS favors differentiation of Th effector cells when cooperates with appropriate activation stimuli such as CD3+CD28 or CD3+IL-2, whereas it supports differentiation of regulatory T cells when costimulatory signals are insufficient, This work was partially supported by Telethon grant E1170 (Rome), FISM grant 2003/R/20 (Genoa), PRIN Project (MIUR, Rome), Fondazione Cariplo (Milan), Compagnia di San Paolo (Turin), Fondazione Cassa di Risparmio di Cuneo (Cuneo), Regione Piemonte (Turin), and Associazione “Amici di Jean” (Turin)

The Fas death receptor is expressed by activated lymphocytes and is involved in switching-off the immune response. Its inherited defects cause auto-immune lymphoproliferative syndrome. Impaired Fas function may also play a role in other auto-immune diseases, such as multiple sclerosis and type 1 diabetes mellitus. The aim of this work was to evaluate Fas function in T cells from patients with chronic inflammatory demyelinating polyneuropathy (CIDP). We evaluated Fas-induced apoptosis in T-cell lines from 27 patients with CIDP, 12 patients with acute inflammatory demyelinating polyneuropathy (AIDP), and 110 controls. CIDP patients displayed lower Fas function than both AIDP patients and controls, whereas no statistically significant difference was found between AIDP patients and controls. Moreover, Fas function was lower in CIDP patients with progressive course than in those with relapsing-remitting course and lower in CIDP patients with axonal damage than in those with pure demyelination. These data suggest that defective Fas function favours CIDP development and aggressive evolution.

Ultraviolet-B (UV-B) radiation (280–320 nm) is an environmental challenge affecting a number of metabolic functions through the generation of reactive oxygen species (ROS). Plants protect themselves from this harmful radiation by synthesizing flavonoids, which act as a screen inside the epidermal cell layer, and by making adjustments to the antioxidant systems at both cell and whole organism level. This study describes the flavonoid content, the photosynthetic pigment composition and the proline, tocopherol and ascorbate content in UV-B exposed maize plants. Following exposure, the tocopherol content was slightly, but significantly lower, pointing to the membrane environment as a primary target for UV-B radiation. The water-soluble antioxidant content was largely unaffected, but an enhanced turnover in the ascorbate–glutathione cycle might be needed for tocopherol regeneration.

Summary The composition of low-molecular-weight thiols in Zea mays L. was investigated by HPLC separation following derivatisation with SBD-F (ammonium 7-fluoro 2,1,3-benzooxadiazole-4-sulfonate) as a fluorescent label. This method was applied to describing variations in the acid-soluble thiol pool in a normal dark/light cycle, and with supplemental UV-B radiation causing moderate photo-oxidative stress. We provide evidence of an increase in leaf η-glu-cys content during the night. This finding suggests that η-glu-cys accumulation in the absence of light is a general phenomenon, though it is not yet clear whether glycine availability from photorespiration is the underlying mechanism in C4 plants too. UV-B radiation caused a significant increase in η-glu-cys content in both shoots and roots. A faster loss of GSH in shoots, in experiments where the glutathione biosynthesis inhibitor buthionine sulfoximine was used, indicates that UV-B radiation enhances glutathione degradation. This is consistent with a higher content of cys-gly (reported to be a η-glutamyl-transpeptidase degradation product of glutathione or its conjugates) in shoots of UV-B exposed seedlings.

In order to examine the light-modulation of nitrate reductase (NR ; EC 1.6.6.1) activity in tissues of maize (Zea mays L. cv. Plauto), the enzyme was assayed under both selective (5 mM Mg 2+ ) and unselective conditions (2 mM EDTA ; V max ), in leaf and root extracts from seedlings transferred from high irradiance (HI ; 450 μmol m- 2 s-' PAR) to shade (200 μmol m- 2 s -1 ) or to darkness. The corresponding changes in tissue nitrate were also evaluated. The results show that maize leaf NR activity (NRA) is subjected to reversible light-modulation involving rapid changes in the sensitivity of the enzyme to inhibition by Mg 2+ . In fact, NR assayed in the presence of Mg 2+ lost about 60% and 20% of its activity after 30 min in the dark and in the shade, respectively. V max started to decrease only after 1-3 h, depending on the extent of light reduction. Deactivation in the shade was reversed in vivo by returning the plants to HI, the kinetics of reactivation being similar to those of deactivation. Following dark-deactivation in vivo, spontaneous reactivation of leaf NR occurred during in vitro incubation at 25°C, and the process was accelerated by AMP. The NR from HI-leaves was deactivated (half-time 2 min) by incubation with MgATP, but the extent of the in vitro deactivation was small. Maize root NR was more sensitive to Mg 2+ than leaf NR, with 5 mM Mg 2+ causing a 50% inhibition of the activity in HI-roots. In contrast to leaves, marked changes in the Mg 2+ -sensitivity of maize root NRA became apparent only after 3 h in the dark or in the shade, when V max had also declined significantly.

The effects of decreased irradiance on fresh and dry weight, root respiration, levels of carbohydrates and N-compounds, and extractable activities of enzymes involved in C and N metabolism were evaluated in maize (Zea mays L. cv. Plauto) seedlings during the 7 days following transfer from 450 to 200 μmol m−2 s−1 PAR. The fresh weight of roots and stems, the initiation of new leaves, root respiration rate, and the accumulation of dry matter, soluble sugars, starch, malate and amino acids in both leaves and roots were strongly reduced at low irradiance. In contrast, the level of nitrate was increased in leaves and only marginally affected in roots. Leaf phosphoenolpyruvate carboxylase (EC 4.1.1.31) activity started to decrease after 24–34 h, whereas ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) activity and chlorophyll content were unaffected or only slightly reduced. In both leaves and roots, the adjustment of N metabolism to low irradiance occurred through a relatively rapid (30% after 10 h) and large (60% after 3 days) decrease of nitrate reductase (NR; EC 1.6.6.1) activity, followed by slower and smaller changes in the activity of nitrite reductase (EC 1.7.7.1), glutamine synthetase (EC 6.3.1.2) and NAD-dependent glutamate dehydrogenase (EC 1.4.1.2). We suggest that the preferential decrease of NR activity relative to other N-assimilating enzymes may be important for preventing the accumulation of toxic N-compounds like ammonia in both leaf and root tissues.

The developmental profile of the activities of some enzymes involved in malate metabolism, namely phosphoenolpyruvate carboxylase (PEPC; EC 4. 1. 1. 31), NAD+-linked (EC 1. 1. 1. 37) and NADP+-linked (EC 1. 1. 1. 82) malate dehydrosenase (MDH), NAD+linked (EC 1. 1. 1. 39) and NADP+-linked (EC 1. 1. 1. 40) malic enzyme (ME), has been determined in leaves of peach [Prunus persica (L.) Batsch cv. Maycrest], a woody C3 species. In order to study the role of these enzymes, their activities were related to developmental changes of photosynthesis, respiration, and capacity for N assimilation. Activities of PEPC, NAD(P)+-MDH and NADP+-ME were high in young expanding leaves and decreased 2- to 3-fold in mature ones, suggesting that such enzymes play some role during the early stages of leaf expansion. In leaves of peach, such a role did not seem to be linked to C3 photosynthesis or nitrate assimilation, in that photosynthetic O2 evolution and activities of nitrate reductase (EC 1. 6. 6. 1) and glutamine synthetase (EC 6. 3. 1. 2) increased during leaf development. In contrast, leaf respiration strongly decreased with increasing leaf age. We suggest that in expanding leaves of this woody species the enzymes associated with malate metabolism have anaplerotic functions, and that PEPC may also contribute to the recapture of respiratory CO2.

γ-Glutamyl transferases (GGT; EC 2.3.2.2) are glutathione-degrading enzymes that are represented in Arabidopsis thaliana by a small gene family of four members. Two isoforms, GGT1 and GGT2, are apoplastic, sharing broad similarities in their amino acid sequences, but they are differently expressed in the tissues: GGT1 is expressed in roots, leaves, and siliques, while GGT2 was thought to be expressed only in siliques. It is demonstrated here that GGT2 is also expressed in wild-type roots, albeit in very small amounts. GGT2 expression is enhanced in ggt1 knockout mutants, suggesting a compensatory effect to restore GGT activity in the root apoplast. Supplementation with 100 μM glutathione (GSH) resulted in the up-regulation of GGT2 gene expression in wild-type and ggt1 knockout roots, and of GGT1 gene expression in wild-type roots. Glutathione recovery was hampered by the GGT inhibitor serine/borate, suggesting a major role for apoplastic GGTs in this process. These findings can explain the ability of ggt1 knockout mutants to retrieve exogenously added glutathione from the growth medium.

Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyneuropathy (CIDP) are immune-mediated neuropathies. GBS is characterized by acute onset and subsequent remission of symptoms, whereas CIDP displays slow progression over at least 2 months. However, a small proportion of CIDP patients display acute onset CIDP (a-CIDP) resembling that of GBS. The Fas receptor is involved in shutting off the immune response and its defective function predisposes to auto-immune diseases. In CIDP patients, Fas function is lower than in GBS patients and healthy controls. This study is aimed at assessing whether evaluation of T-cell Fas function helps in early discrimination between GBS and a-CIDP. Fas function was evaluated in patients with acute onset polyneuropathy: 55 retrospective patients analyzed after development of GBS or a-CIDP before year 2005; 50 prospective patients analyzed at onset after year 2005 and followed up for development of GBS or a-CIDP. In both groups, a-CIDP patients displayed defective Fas function, whereas GBS patients displayed normal function. These findings suggest that the evaluation of Fas function in acute onset polyneuropathy helps in early prediction of long-term outcome.

12 páginas -- PAGS nros. 141-152, Objective: Cell infection by HIV-1 is inhibited by both the expression of CD38 and a soluble peptide (sCD38p) corresponding to its extracellular membrane-proximal amino acid sequence (amino acids 51 - 74). We show here the effects of PEG conjugation to sCD38p and provide new insights into the mechanisms behind the anti-HIV-1 effects of CD38 and derived peptides. Research design/methods: In-vitro and in-silico study. Results: PEGylation of sCD38p increased its ability to inhibit replication of HIV-1 in MT-4 cells and syncytia formation in cocultures of MT-2 and persistently HIV-1IIIB-infected H9IIIB cells. In silico modeling suggests that sCD38p and CD4 form stable heterodimers involving, among others, an interaction between lysine 57 (K57) of CD38 and a groove in the CD4 receptor, which, in CD4/gp120 complexes, is partially occupied by a lysine residue of the HIV-1 envelope glycoprotein. K57 substitution with a glycine in sCD38p impaired its ability to inhibit syncytia formation in MT-2/H9IIIB cell cocultures and gp120 binding to CD4 in a mouse T cell line expressing human but not mouse CD4. Conclusions: PEGylation significantly improves the anti-HIV-1 activity of sCD38p, whose effect is probably due to competition with gp120 for a common binding site on CD4 although other mechanisms cannot be excluded so far. The inhibitory concentrations of the sCD38p-PEG as well as its poor toxicity, merit further consideration in anti-HIV-1 strategies

Objective Hereditary periodic fever syndromes (HPFs) develop as a result of uncontrolled activation of the inflammatory response, with a substantial contribution from interleukin-1β or tumor necrosis factor α (TNFα). The HPFs include familial Mediterranean fever (FMF), hyperimmunoglobulinemia D with periodic fever syndrome (HIDS), TNF receptor–associated syndrome (TRAPS), and cryopyrinopathies, which are attributable to mutations of the MEFV, MVK, TNFRSF1A, and CIAS1 genes, respectively. However, in many patients, the mutated gene has not been determined; therefore, the condition in these patients with an HPF-like clinical picture is referred to as idiopathic periodic fever (IPF). The aim of this study was to assess involvement of X-linked inhibitor of apoptosis (XIAP), which plays a role in caspase inhibition and NF-κB signaling, both of which are processes that influence the development of inflammatory cells. Methods The XIAP gene (X-linked) was sequenced in 87 patients with IPF, 46 patients with HPF (13 with HIDS, 17 with TRAPS, and 16 with FMF), and 182 healthy control subjects. The expression of different alleles was evaluated by sequencing XIAP-specific complementary DNA mini-libraries and by real-time polymerase chain reaction and Western blot analyses. The functional effect of XIAP on caspase 9 activity was assessed by a fluorimetric assay, and cytokine secretion was evaluated by enzyme-linked immunosorbent assay. Results Sequencing disclosed a 1268A>C variation that caused a Q423P amino acid substitution. The frequency of 423Q-homozygous female patients and 423Q-hemizygous male patients was significantly higher in the IPF group than in the control group (69% versus 51%; odds ratio 2.17, 95% confidence interval 1.23–3.87, P = 0.007), whereas no significant difference was detected in the HPF group (59%) compared with controls. In primary lymphocytes and transfected cell lines, 423Q, as compared with 423P, was associated with higher XIAP protein and messenger RNA expression and lower caspase 9 activation. In lipopolysaccharide-activated monocytes, 423Q was associated with higher secretion of TNFα. Conclusion These results suggest that 423Q is a predisposing factor for IPF development, possibly through its influence on monocyte function.

S New Biotechnology · Volume 25S · September 2009 olus vulgaris L.) is not known. Using immunolocalization, in some species such as Zea mays, Hordeum vulgare and Arabidopsis thaliana Sus showed expression in the basal endosperm transfer cells and in developing embryos. In order to better define the role of Sus in Phaseolus, we have compared the pattern expression of Sus during developmental progression of wild-type and an ethyl methanesulfonate (EMS) mutant embryos. Homozygous embryos isolated from mutant aborted pods were arrested at the globular stage; they present a developmental delay during embryogenesis and lethality in seed growth. Expression profiling of Sus by reverse transcriptase polymerase chain reaction (RT-PCR) showed that the gene was active at low levels in vegetative wild-type tissues such as leaves, flowers, cotyledons and roots. Highest level of expression was observed in seeds and nothing in stems. In situ hybridization studies in wild-type, 3 days after pollination (DAP), show the gene activity in endosperm, suspensor, embryo, but also in a group of cells located in the outer and inner integumentary layer. At 7 and 8 DAP Sus gene product mRNA was detected specifically in suspensor and no signal detect in other tissue. However, at 9 and 12 DAP Sus expression was revealed in suspensor, endothelium layer, and cotyledons, but was absent from the endosperm, outer and inner integumentary layer. These temporal changes in cellular localization of Sus gene during common bean seed development are comparable with those reported in Arabidopsis seed. These data suggested a possible role of Sus in carbon portioning during early to mid stages of seed development, as a process to maintain sink strength. In later stage, Sus play a role within the cotyledon in sucrose utilization. In the EMS mutant plant, disruption in Sus pattern and expression highlight the roles of Sus gene during seed development in common bean. doi:10.1016/j.nbt.2009.06.965

Gamma-glutamyl transferase/transpeptidase (GGT, (5-l-glutamyl)-peptide:amino-acid 5-glutamyltransferase; EC 2.3.2.2.) is an ectoenzyme promoting the cleavage of the gamma-glutamyl moiety of glutathione (GSH) and gamma-glutamyl related compounds. In this work, we describe the localization of GGT by enzymehistochemical and immunohistochemical analysis in maize plants. Our results show that the tissue spatial distribution of GGT activity closely correlates with the localization of the GGT protein. We also demonstrate that GGT activity and protein are unevenly distributed in tissues, being higher in the epidermis and stomata, parenchyma of conductive elements and root meristem. These results can contribute to our understanding of GGT function and regulation as well as its role in glutathione metabolism. To date, these are largely unknown in plants.

Mutations decreasing function of the Fas death receptor cause the autoimmune lymphoproliferative syndrome (ALPS) with autoimmune manifestations, spleen/lymph node enlargement, and expansion of CD4/CD8-negative T cells. Dianzani Autoimmune Lymphoproliferative Disease (DALD) is a variant lacking this expansion. Perforin is involved in cell-mediated cytotoxicity and its biallelic mutations cause familial hemophagocytic lymphohistiocytosis (HLH). We previously described an ALPS patient carrying heterozygous mutations of the Fas and perforin genes and suggested that they concurred in ALPS. This work extends the analysis to 14 ALPS, 28 DALD, and 816 controls, and detects an N252S amino acid substitution in 2 ALPS, and an A91V amino acid substitution in 6 DALD. N252S conferred an OR = 62.7 (P = .0016) for ALPS and A91V conferred an OR = 3 (P = .016) for DALD. Copresence of A91V and variations of the osteopontin gene previously associated with DALD conferred an OR = 17 (P = .0007) for DALD. In one N252S patient, NK activity was strikingly defective in early childhood, but became normal in late childhood. A91V patients displayed lower NK activity than controls. These data suggest that perforin variations are a susceptibility factor for ALPS/DALD development in subjects with defective Fas function and may influence disease expression.

A large human nonimmune phage antibody library was screened by affinity chromatography to select single-chain antibodies directed against the human receptor tyrosine kinase (RTK) Ron. As antigen, we used a GST fusion protein (GST-IRP − ) containing the whole intracellular portion of Ron except for the carboxyl-terminal arginine–proline-rich motif. One selected phage was highly specific for Ron when tested in an enzyme-linked immunosorbent assay (ELISA). We report here the immunological characterization of this anti-Ron single-chain antibody (sc7) and show that it recognizes both denatured and native forms of the receptor. The epitope bound by sc7 maps within the first 50 amino acid residues of the juxtamembrane domain of Ron. This monoclonal fragment does not cross-react with other receptor tyrosine kinases including the closely related human proto-oncogene Met. We demonstrate that the isolated antibody fragment interacts in vivo with the intracellular domain of Ron in mammalian cells.

The effect of UV-B radiation on FW, leaf and stem length, photosynthetic O2 evolution, levels of carbohydrates and nitrates, and extractable activities of some of the enzymes involved in C and N metabolism was evaluated in barley (Hordeum vulgare L. cv. Express) seedlings during the 9 days following transfer to an UV-B enriched environment. The results show that under our experimental conditions UV-B radiation scarcely affects the photosynthetic competence of barley leaves, expressed as RuBP carboxylase (EC 4.1.1.39) activity, O2 evolution rate and chlorophyll content. Nevertheless, this treatment induced significant alterations of the enzyme activity of nitrate reductase (EC 1.6.6.1) and glutamine synthetase (EC 6.3.1.2), although only after a few days of treatment. The effects were not confined to the exposed tissue, but were detectable also at the root level. In fact, nitrate reductase decreased in response to UV-B in both leaf and root tissue, whereas glutamine synthetase was affected only in the root. In contrast, nitrate content was not influenced by the treatment, neither in root nor in leaf tissue, whilst leaf sucrose diminished in exposed plants only on the last day of treatment.

The activities of enzymes involved in ammonia metabolism ferredoxin-dependent glutamate synthase (Fd-GOGAT), glutamine synthetase (GS) and glutamate dehydrogenase (GDH), the rates of photosynthetic oxygen evolution, dark respiration, and the activity of RuBP carboxylase (RuBPC) were determined in alfalfa (Medicago sativa L.) leaves taken from the apex (apical leaves), from the second to the fourth internode (mature leaves) and from the bottom of the canopy (basal leaves). Photosynthetic rate and the activities of RuBPC, GS and Fd-GOGAT showed their maximum in the mature leaves. The respiration rate together with amino acid and ammonium contents decreased with leaf age, whereas the opposite was true for GDH activity. Basal leaves still maintained substantial levels of chlorophylls, GS and Fd-GOGAT activities and oxygen evolution rate, thus suggesting that photosynthesis has some role in the reassimilation of the nitrogen liberated during protein degradation.

The growth of the whole plant and the chlorophyll content, oxygen evolution, and chloroplast ultrastructure of leaf tissues have been studied in maize plants grown on a culture medium either without cadmium (Cd) or supplied with increasing concentrations of the metal. The plants treated with high Cd concentrations showed symptoms of heavy metal toxicity, such as length reduction of both roots and shoots, leaf bleaching, ultrastructural alterations of chloroplasts and lowering of photosynthetic activity. Some symptoms appeared at 100 μM Cd, but the strong toxic effects of the metal were found only at 250 μM Cd.

Attraverso un dettagliato studio del disegno tratto da Stefano Orlandi nel 1715 da polittico eseguito da Agostino de' Marchi e dipinto da Francesco del Cossa e Ercole Roberti per l'altare di Floriano Griffoni in San Petronio a Bologna e della lettera che gli si accompagna, viene proposta una più plausibile disposizione degli elementi sopravvissuti alla sua distruzione.

Edizione del documento di commissione della pala del Martirio di San Sebastiano di Federico Barocci, stilato dal notaio urbinate Giulio Corvini il 9 novembre 1557 su mandato del committente dell'opera Benedetto Bonaventura.