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• The shedder status influences the extent of DNA transfer. • The order in which participants handled an object did not influence DNA quantity. • Secondary DNA transfer by working gloves onto a screwdriver is possible. • We observed innocent persons' DNA at the 'crime scene´. With the development of highly sensitive STR profiling methods, combined with sound statistical tools, DNA analysis on the (sub-)source level is hardly ever seriously questioned in court. More often, the exact mode of DNA transfer to the crime scene is questioned. In burglary cases, in particular when gloves are worn, secondary DNA transfer is often discussed as explanation for finding a DNA profile matching the accused because it is well known that gloves can act as a potential vector for indirect DNA transfer. In this study we investigated the shedder status as a possible factor influencing the extent of secondary DNA transfer to a crime scene, with the person committing the crime wearing working gloves. Firstly, the shedder status for 40 participants (20 male, 20 female) was determined, following a previously published procedure. Good shedders (n = 12) were found to deposit a higher amount and quality of DNA onto objects, compared to bad shedders (n = 25). Secondly, participants were paired into four groups (good with good; good with bad; bad with good; bad with bad), each group consisting of five pairs. The first participant (P1) of each pair used working gloves to pack and carry a box to simulate a house move. Two days later, the second participant (P2) of the pair wore the same pair of gloves to simulate a burglary, using a screwdriver as a break-in tool. After taking swabs of the outside and inside of a glove (primary DNA transfer) and the handle of the screwdriver (secondary DNA transfer), full DNA analysis was performed. Our experiments show that good shedders, overall, deposit more DNA than bad shedders, both onto the outside and the inside of the glove, regardless of being P1 or P2. When conducting the experiments with two participants sharing the same shedder status, no significant differences occurred in the number of deposited alleles. In six out of 19 cases a DNA profile matching P1 was found (binary LR>106) on the screwdriver and in all six cases P1 was a good shedder. Our results indicate that the shedder status of an individual affects the extent of DNA transfer. They further confirm the possibility of an innocent person's DNA profile being found on an object they never handled. [ABSTRACT FROM AUTHOR]

Linear-after-the-exponential (LATE)-PCR describes a novel approach to asymmetric PCR which uses adjusted melting temperatures of the limited primer to increase PCR efficiency. In this proof-of-principle study we show that linear amplification is possible over a wide range of amplification cycles. The curve characteristics of the real time PCR show that the plateau effect of conventional PCR can be avoided by this innovative technique. [ABSTRACT FROM AUTHOR]

Pro-inflammatory chemokines and their receptors exhibit elementary functions in cell migration and in Th1-driven inflammatory conditions. One therapeutic strategy to prevent accumulation of pro-inflammatory immune cells is the use of specific chemokine receptor antagonists. An interesting and promising candidate in this context is the viral antagonist MIP-II (vMIP-II) that acts on a broad spectrum of chemokine receptors. To study the in vitro and in vivo effects of vMIP-II on pro-inflammatory chemokine receptor function, we further characterized an ovalbumin-specific murine central memory Th1IF12 clone by using RT-PCR, cDNA array and cytometry. Using in vitro chemotaxis assays we show that eukaryotically generated vMIP-II strongly inhibited migration of CCL2- or CCL5-stimulated Th1 IF12 cells. Using intravital microscopy, we observed that CCL5 induced rolling of Th1 cells in the ear vasculature of C57Bl/6 mice. Pre-treatment with vMIP-II significantly reduced CCL5-induced rolling of Th1 cells to basal levels, indicating, that vMIP-II is also active in vivo (proportion of rolling cells: 19.4 +/- 3.8%, 39.8 +/- 2.9% and 26.1 +/- 3.2%). In addition, investigating the anti-inflammatory action of vMIP-II in adoptive transfer of immunity and dinitrofluorobenzene-induced cutaneous hypersensitivity reaction using C57Bl/6 mice, we show a direct inhibitory effect of vMIP-II on the sensitization phase [Delta ear swelling 62 and 37 cm x 10(-3) for controls and vMIP-II treated mice (2.5 mg/kg), respectively] and effector phase (Delta ear swelling 14.8 and 3.6 cm x 10(-3) for controls and vMIP-II treated mice (2.5 mg/kg), respectively) of cutaneous hypersensitivity. These data indicate that vMIP-II is a promising agent to interfere with chronic inflammatory (skin) diseases.