Synthesis and biological evaluation of 3-hydroxymethyl-5-(1H-1,2,3-triazol) isoxazolidines

A synthetic approach towards a series of 3-hydroxymethyl-5-(1H-1,2,3-triazol)isoxazolidines has been reported, according to a procedure based on the cycloaddition reaction, under microwave irradiation, of a nitrone with 1-vinyl triazoles, prepared by a click reaction of azides with alkynes. Biological tests show that the synthesized compounds are able to inhibit proliferation of follicular and anaplastic human thyroid cancer cell lines, with IC₅₀ values ranging from 3.87 to 8.76 μM. The obtained compounds induce caspase-3 activation and DNA fragmentation prevalently in follicular human thyroid cancer cell lines.     

A marine/terrestrial integration for mid-late Holocene vegetation history and the development of the cultural landscape in the Po valley as a result of human impact and climate change

Integration of pollen data from both marine and terrestrial cores contributes to the understanding of the timing of the climatic and human forces that shaped the cultural landscapes in the Italian peninsula. This paper focuses on the relation between natural and human landscapes, and the development of the cultural landscape from the Bronze Age to the medieval period and modern times. Two records were studied within independent projects, first the marine core RF93-30, from the central Adriatic, with a sediment source area including the Po valley and which spans the last 7,000?years, and secondly, material from the site of Terramara di Montale, a Bronze Age settlement on the Po plain, which was occupied from approximately 3550–3200?cal. b.p. The original chronology of the marine core was developed by using the magnetic inclination of the secular variation record and two ¹⁴C dates carried out on benthic and planktic foraminifera at depths of 527 and 599?cm. Its pollen record shows a gradual irreversible trend towards increasing aridity since 5700?cal. b.p. and, just after around 5100?cal. b.p., a Picea decline and a Quercus ilex type increase indicate less cool conditions. Human impact introduces rapid changes, such as the decrease of Abies alba, thinned by the reduction of precipitation and further cleared before or during the Early Bronze Age, followed by the fall of oaks. The latter started after around 3900?cal. b.p., and became evident at around 3600?cal. b.p. The gradual increase in signs of open landscape and woodland clearance correspond to the onset of Middle Bronze Age settlements in the Po valley, and to the development of the cultural landscape in the region. The impact of the terramare people includes woodland management by coppicing, and division of the territory into a patchwork of pastures and fields. Dry environments are indicated mainly by Cichorioideae, resulting from the continued human pressure, and these spread since the Recent Bronze Age. Of the possible causes for the decline of the terramare, we suggest that climate would have been less important in the decline than in the onset phases. The later cultural landscapes are mainly indicated by the trends of the Olea, Juglans and Castanea (OJC) records, besides those of cereals. At around 700?cal. b.p., the “chestnut landscape” spread while modern times are shown by the finds of Zea mays.     

Phosphoglucomutase is absent in Trypanosoma brucei and redundantly substituted by phosphomannomutase and phospho-N-acetylglucosamine mutase

The enzymes phosphomannomutase (PMM), phospho‐N‐acetylglucosamine mutase (PAGM) and phosphoglucomutase (PGM) reversibly catalyse the transfer of phosphate between the C6 and C1 hydroxyl groups of mannose, N‐acetylglucosamine and glucose respectively. Although genes for a candidate PMM and a PAGM enzymes have been found in the Trypanosoma brucei genome, there is, surprisingly, no candidate gene for PGM. The TbPMM and TbPAGM genes were cloned and expressed in Escherichia coli and the TbPMM enzyme was crystallized and its structure solved at 1.85 Å resolution. Antibodies to the recombinant proteins localized endogenous TbPMM to glycosomes in the bloodstream form of the parasite, while TbPAGM localized to both the cytosol and glycosomes. Both recombinant enzymes were able to interconvert glucose‐phosphates, as well as acting on their own definitive substrates. Analysis of sugar nucleotide levels in parasites with TbPMM or TbPAGM knocked down by RNA interference (RNAi) suggests that, in vivo, PGM activity is catalysed by both enzymes. This is the first example in any organism of PGM activity being completely replaced in this way and it explains why, uniquely, T. brucei has been able to lose its PGM gene. The RNAi data for TbPMM also showed that this is an essential gene for parasite growth.     

Inaccuracies in self-reported intake identified by comparison with the doubly labelled water method

To test the accuracy of self-reported energy intake, reported intake was compared with measured energy expenditure. Results from nine studies were reviewed in which intake data were obtained by recall or weighed record for at least 7 days. Expenditure was measured for 7 days or more by the doubly labelled water method. Individual differences between reported intake and expenditure were large (range +25 to -76%). Group mean differences were smaller. Lean, nonathletic groups living in industrialized countries demonstrated the smallest mean difference between self-reported energy intakes and expenditure (0 to -20%). Obese populations demonstrated the largest mean differences (-35 and -50%), but women living in the Gambia and elite athletes also demonstrated large mean differences. Most of the difference appears to be due to under-reporting, but some subjects lost weight during the reporting period indicating that some of the difference was due to undereating. Because the greatest bias was observed in obese subjects, current methods for self-reported energy intake are not recommended for use in obesity research.     

Pollen grains in human cytology

This paper reports on pollen analyses carried out in the course of a ten-year investigation, on many thousands of cytological smears coming from various organs and systems of the human body, and prepared for diagnostic purposes. The frequency and the significance of the pollen records vary according to the specific cytological field taken into account. In the urinary sediment smears, nipple secretions, and needle aspirations the polliniferous smears are very few, and the pollen number per smear is low (max 14 pollen grains, belonging always or mostly to anemophilous species). In these cases, the pollen records evidence the airborne contamination during medical procedures, the same happens with most of cervico-vaginal smears. In some cervico-vaginal smears, the high frequency of pollen grains belonging to pharmaceutical taxa suggests that lavages with vegetable components were used by the patients before undergoing the test. In nasal, bronchial and conjunctival cytology greater amounts of polliniferous slides were recorded (in bronchial/nasal cytology also a higher number of pollen grains per smears, up to 114–428 respectively) and pollen spectra reflected the vegetational environment of the patients' living sites. In these cases, most pollen grains are thought to be really present on mucosae when the samples were taken. In these cytological fields pollen analysis may be useful for diagnostic purposes, above all in case of allergic pathology to detect the pollen grains causing the disease.     

New adaptive chiral thiophene ligands for copper-catalyzed asymmetric Henry reaction

A new type of adaptive chiral thiophene-based ligands 3 has been designed and developed. The synthetic flexibility of the thienyl ring allowed for the preparation of polydendate C2-symmetric ligands in good yields, carrying simultaneously "hard" as well as "soft" coordinating atoms (i.e., N, S). The coordination attitude of 3 was then tested in the enantioselective base-free Cu(OAc)(2)-catalyzed addition of nitromethane to aromatic aldehydes, leading to the corresponding beta-nitro alcohols in excellent yields and enantiomeric excesses up to 86%.     

Antibodies designed as effective cancer vaccines

Antigen/antibody complexes can efficiently target antigen presenting cells to allow stimulation of the cellular immune response. Due to the difficulty of manufacture and their inherent instability complexes have proved inefficient cancer vaccines. However, anti-idiotypic antibodies mimicking antigens have been shown to stimulate both antibody and T cell responses. The latter are due to T cell mimotopes expressed within the complementarity-determining regions (CDRs) of antibodies that are efficiently presented to dendritic cells in vivo. Based on this observation we have designed a DNA vaccine platform called ImmunoBody™, where cytotoxic T lymphocyte (CTL) and helper T cell epitopes replace CDR regions within the framework of a human IgG1 antibody. The ImmunoBody™ expression system has a number of design features which allow for rapid production of a wide range of vaccines. The CDR regions of the heavy and light chain have been engineered to contain unique restriction endonuclease sites, which can be easily opened, and oligonucleotides encoding the T cell epitopes inserted. The variable and constant regions of the ImmunoBody™ are also flanked by restriction sites, which permit easy exchange of other IgG subtypes. Here we show a range of T cell epitopes can be inserted into the ImmunoBody™ vector and upon immunization these T cell epitopes are efficiently processed and presented to stimulate high frequency helper and CTL responses capable of anti-tumor activity.Key words: DNA vaccines, cancer vaccines, melanoma, CTL, helper T cells     

Cumulative Risk of Bovine Mastitis Treatments in Denmark,Finland, Norway and Sweden

Data from the national dairy cow recording systems during 1997 were used to calculate lactation-specific cumulative risk of mastitis treatments and cumulative risk of removal from the herds in Denmark, Finland Norway and Sweden. Sweden had the lowest risk of recorded mastitis treatments during 305 days of lactation and Norway had the highest risk. The incidence risk of recorded mastitis treatments during 305 days of lactation in Denmark, Finland, Norway and Sweden was 0.177, 0.139, 0.215 and 0.127 for first parity cows and 0.228, 0.215, 0.358 and 0.204 for parities higher than three, respectively. The risk of a first parity cow being treated for mastitis was almost 3 times higher at calving in Norway than in Sweden. The period with the highest risk for mastitis treatments was from 2 days before calving until 14 days after calving and the highest risk for removal was from calving to 10 days after calving in all countries.The study clearly demonstrated differences in bovine mastitis treatment patterns among the Nordic countries. The most important findings were the differences in treatment risks during different lactations within each country, as well as differences in strategies with respect to the time during lactation mastitis was treated.