The recovery of pollen evidence from documents and its forensic implications.

Three experiments were undertaken to establish the potential for forensic palynological analysis in cases of suspected document fraud. The first study tested 6 different types of paper and 9 different types of ink (n=54) and it was established that the best retainer of particulates (in this case a proxy was used in the form of UV powder) was medium biro ink and Wove and Connoisseur paper. It was found that for the different paper types 42-52% of the particulates collected were found in the ink and thus both the paper and the ink are potentially valuable sources of trace evidence in a forensic investigation. The second study sought to address the differences in the spatial distribution of particulates on documents when writing took place before or after the paper was treated with UV particulates. Ninety-six observations were made for each piece of paper tested and it was found that when the writing took place after the particulates were applied to the paper; more particulates were retained on the paper in contrast to when the writing took place before the particulate treatment. The spatial distribution of particulates was also affected, with particulates being retained in the folds of the paper when the writing took place before particulate treatment in contrast to a more erratic pattern that emerged due to the pressure of the hand of the writer when the writing took place after the particulate treatment. The third study utilised lily (Lilium) pollen grains and the findings broadly concurred with the second study. The main difference identified was when the writing took place before the particulates were applied; when UV powder was used the particulates were retained in the folds of the paper whereas this pattern was not seen to the same degree when pollen grains were used due to their 'stickier' nature. Envelopes and the pen nibs were also found to be rich sources of pollen grains after the experiments were undertaken. These studies have implications for the application of forensic palynology in cases of suspected document fraud. Pollen grains may well be present, and their analysis has the potential to reveal not only the timing of the generation of the document, but the spatial trends revealed indicate that it may well be possible to establish the sequence of significant events for forensic reconstruction. As such forensic palynology is demonstrated to have great potential in aiding forensic investigations, and is as yet an under-utilised form of trace evidence.

The functional logic of cytosolic 5'-nucleotidases.

Adenosine- and uridine-cytidine kinases, purine-nucleoside phosphorylase, hypoxanthine-guanine phosphoribosyl transferase, and several related enzymes, are components of the salvage pathways which reduce the loss of intracellular purine and pyrimidine rings. Although this could explain the role of these enzymes, it poses a problem of the role of the cytosolic 5'-nucleotidase. Why are nucleosides produced from nucleoside-monophosphates, only to be converted back to the same compounds? To date, it is well established that a cross talk exists between the extracellular and intracellular nucleoside metabolism. In districts, such as brain, which are dependent on salvage nucleotide synthesis, nucleosides are produced through the action of the ecto-5'-nucleotidase, the last component of a series of plasma-membrane bound enzyme proteins, catalyzing the successive dephosphorylation of released nucleoside-triphosphates. Both nucleosidetriphosphates (mainly ATP and UTP) and nucleosides (mainly adenosine), act as extracellular signals. Once transported into cell cytosol, all nucleosides are salvaged back to nucleoside-triphosphates, with the exception of inosine, whose salvage is limited to IMP. Intracellular balance of nucleosides is maintained by the action of several enzymes, such as adenosine deaminase, uridine phosphorylase and cytidine deaminase, and by at least three 5'-nucleotidases, the ADP activated AMP preferring cN-IA, the ATP-ADP activated IMP-GMP preferring cN-II, and the UMP-CMP preferring cN-III. Here we are reviewing the mechanisms whereby cytosolic 5'-nucleotidases control changes in nucleoside and nucleotide concentration, with the aim to provide a common basis for the study of the relationship between biochemistry and other related disciplines, such as physiology and pharmacology.

VEGF inhibition and cytotoxic effect of aplidin in leukemia cell lines and cells from acute myeloid leukemia.

BACKGROUND: Aplidine (APL) is a marine depsipeptide isolated from the Mediterranean tunicate Aplidium albicans that is under clinical phase II development. In contrast to the lack of bone marrow toxicity reported in phase I/II studies, it has been shown to induce cytotoxicity at very low concentration against lymphoblastic leukemia blast, as well as having an impact in the vascular endothelial growth factor (VEGF)/VEGF receptor 1 loop. PATIENTS AND METHODS: To confirm these findings we investigated APL-related VEGF inhibition and its cytotoxic effect on myeloid leukemic cells lines (K-562, HEL and HL60) and fresh leukemia blasts derived from 30 patients with acute myeloid leukemia (AML). The conventional active 4-demetoxi-daunorubicin (idarubicin; IDA) was included as a positive control. RESULTS: APL was found to be significantly (P<0.001) more active than IDA in obtaining 50% growth-inhibition in K-562, HEL and HL60 cell lines. Results obtained with AML blast cells were super imposible. ID(50) ranged from 0.024 to 0.610 microM for IDA (0.200+/-0.176) and from 0.001 to 0.108 microM for APL (0.020+/-0.031). Annexin V tests and cell cycle analysis performed on cell lines confirmed the stronger citotoxic capability of APL as apoptotic inducer and as a G(1) blocker. The inhibitory effects of APL on VEGF release and secretion have been confirmed by ELISA tests performed on HEL: the VEGF concentration in cell surnatant was reduced from 169 to 36 pg/ml after 24 h of exposure to a pharmacological concentration of APL. CONCLUSIONS: APL harbors a strong in vitro antileukemic activity at a concentration achievable in patients at non-myelotoxic doses. Our data also support the notion of an impact on VEGF secretion. Clinical studies with this new marine-derived compound in relapsed/resistant leukemia are underway.