Cross-border patterns in DNA matches between the Netherlands and Belgium.

In this article we present the results of a study to explore if cross-border DNA matches between the Netherlands and Belgium are relatively more likely to occur in areas near the Dutch-Belgian border than in areas at some distance from this border. For this study we used the results of the transnational DNA profile exchange and comparison between the Belgian and Dutch DNA databases, which first took place in 2014. It appears that the Dutch regions adjacent to Belgium, i.e., Zeeland-West-Brabant, Oost-Brabant and Limburg, have relatively more DNA matches with Belgium than the other Dutch regions. In other words, a DNA profile obtained from a crime scene close to the Dutch border with Belgium is more likely to match with the profile of a person whose DNA profile is stored in the Belgian database than a DNA profile that originates from a crime scene further afield. Our data suggest that crimes committed by repeat offenders show a spatial pattern despite the presence of a national border, with crime scenes clustering in relatively close proximity to each other. The results of this study provide a better understanding of geographical patterns of cross-border criminal mobility.

Of earprints, fingerprints, scent dogs, cot deaths and cognitive contamination--a brief look at the present state of play in the forensic arena.

Over the last decades, the importance of technical and scientific evidence for the criminal justice system has been steadily increasing. Unfortunately, the weight of forensic evidence is not always easy for the trier of fact to assess, as appears from a brief discussion of some recent cases in which the weight of expert evidence was either grossly over- or understated. Also, in recent years, questions surrounding the value of forensic evidence have played a major role in the appeal and revision stages of a number of highly publicized criminal cases in several countries, including the UK and the Netherlands. Some of the present confusion is caused by the different ways in which conclusions are formulated by experts working within the traditional approach to forensic identification, as exemplified by (1) dactyloscopy and (2) the other traditional forensic identification disciplines like handwriting analysis, firearms analysis and fibre analysis, as opposed to those working within the modern scientific approach used in forensic DNA analysis. Though most clearly expressed in the way conclusions are formulated within the diverse fields, these differences essentially reflect the scientific paradigms underlying the various identification disciplines. The types of conclusions typically formulated by practitioners of the traditional identification disciplines are seen to be directly related to the two major principles underpinning traditional identification science, i.e. the uniqueness assumption and the individualization principle. The latter of these is shown to be particularly problematic, especially when carried to its extreme, as embodied in the positivity doctrine, which is almost universally embraced by the dactyloscopy profession and allows categorical identification only. Apart from issues arising out of the interpretation of otherwise valid expert evidence there is growing concern over the validity and reliability of the expert evidence submitted to courts. While in various countries including the USA, Canada and the Netherlands criteria have been introduced which may be used as a form of input or output control on expert evidence, in England and Wales expert evidence is much less likely to be subject to forms of admissibility or reliability testing. Finally, a number of measures are proposed which may go some way to address some of the present concerns over the evaluation of technical and scientific evidence.

Effects of a vasopressin antagonist in women with dysmenorrhea.

We compared menstrual pain, uterine contractility and blood circulation, and plasma concentrations of vasopressin and prostaglandin F(2alpha) metabolite in women with versus without primary dysmenorrhea, and determined the effects of a vasopressin antagonist, 1-deamino-2-D-Tyr(OEt)-4-Thr-8-Orn-oxytocin (Atosiban), on these parameters. Our results do not support the contention that vasopressin is involved in the etiology of dysmenorrhea, plasma concentrations of vasopressin being similar in dysmenorrheic women and controls, and the vasopressin antagonist Atosiban having no effect on menstrual pain, intrauterine pressure or uterine artery pulsatility index in dysmenorrheic women.

Pharmacokinetics and antidiuretic effect of a new vasopressin analogue (F992) in overhydrated male volunteers.

OBJECTIVE: The aim of the present study was to study the pharmacokinetics, the antidiuretic effects and the safety of [D-Phe2, Thi3, alpha-Me-Abu4, Hyp7, D-Arg8]dC1-vasopressin, a new antidiuretic peptide (F992, Ferring, Sweden), administered as intravenous infusion to orally overhydrated male volunteers. METHODS: Eight healthy male volunteers participated in this open study consisting of two parts: a dose titration study and a safety study. In the dose titration study ascending doses of F992 were administered to volunteers in pairs in order to find a dose that within 1 h after the infusion, in both subjects, caused a reduction of the urine flow rate to below 5 ml x min(-1) (target dose). Subsequently, this target dose was administered to all volunteers. In the safety study the target dose was doubled and given to all volunteers. On each study occasion, in both study parts, the subjects were orally overhydrated with water. F992 was administered as i.v. infusion approximately 1.5 h after the start of the hydration procedure. Throughout the study days, blood was sampled for determination of plasma concentrations of F992 and for safety evaluation. Urine was collected at intervals in order to estimate flow rate and osmolality. RESULTS: The target dose was found to be 4.0 microg as this dose fulfilled the criteria regarding antidiuretic effect, consequently 8.0 microg was administered to all subjects in the safety study. After infusion of 4.0 and 8.0 microg, the median half-lives of elimination were 4.72 (range 3.99-6.53) h and 3.85 (range 3.04-11.08) h, respectively. The plasma clearance and the volume of distribution at steady state were estimated to be 0.88 (SD 0.24) ml x min(-1) x kg(-1) and 326 (SD 68) ml x kg(-1)] after infusion of 4 microg. After the highest dose (8 microg), the corresponding estimates were 0.86 (SD 0.32) ml x min(-1) x kg(-1) and 299 (SD 81) ml x kg(-1), respectively. Significantly (P = 0.033) different maximum mean urine osmolalities were produced after infusion of 4.0 and 8.0 microg of F992 (534 (SD 318) vs 732 (SD 189) mOsmol x kg(-1)). The median times to reach these values showed some tendency to be longer for the highest dose, however statistical significance was not reached. No serious adverse events were observed during the study. CONCLUSION: We found it safe to administer F992 as infusion to overhydrated male volunteers. The results suggest that F992 has a longer half-life and a lower potency than the widely used peptide desmopressin.

The use of reporter antigens in the popliteal lymph node assay to assess immunomodulation by chemicals.

Various drugs and other chemicals can induce T-cell-dependent B-cell activation which may lead to allergic or autoimmune-like diseases. Because the nature of the relevant (neo-) antigens is generally not known and probably depends on the chemical, we have explored the potential use of reporter antigens to determine T-cell-dependent B-cell activation by chemicals. TNP-Ficoll and TNP-OVA were used for this purpose because they are recognized by the same TNP-specific B cells, but these cells require distinct costimulation for specific antibody production. It was found that HgCl2, phenytoin, nitrofurantoin, and D-penicillamine stimulated IgG1 production to both antigens, incomplete Freund's adjuvant, silica, and dimethylsulfoxide to TNP-OVA only, and LPS and hydroxyl-amino procainamide to TNP-Ficoll alone. The diabetogene streptozotocin did not enhance IgG1 production, but may enhance a cellular response instead. Tolerogens and a T-cell antigen without intrinsic adjuvant activity did not influence the responses. The IgG1 production to TNP-Ficoll was local and transient, and did not always require T cells. In contrast, responses to TNP-OVA could be measured in serum, led to specific memory, and were strictly T-cell dependent. These results demonstrate that specific antibody production to reporter antigens indicates immunostimulatory effects of chemicals more sensitive than PLN cell count and provides important mechanistic information. Moreover, with TNP-OVA as reporter antigen the kinetics and regulation of chemically enhanced immune responses can be studied without the need to know the relevant neo-antigens for each individual compound.

Short term pituitary desensitization: effects of different doses of the gonadotrophin-releasing hormone agonist triptorelin.

Gonadotrophin-releasing hormone agonists (GnRHa) are used to prevent inadequate luteinizing hormone (LH) surges during ovarian stimulation in in-vitro fertilization (IVF). Dose studies for optimal dose assessment are lacking and unfavourable effects of the agonist on granulosa function and oocyte quality have been suggested. This double-blind randomized study was undertaken to assess the effect of four different doses to triptorelin on the degree of desensitization of the pituitary, and the recovery time of pituitary function after withdrawal of the agonist. Sixty-six regularly cycling women were allocated to a treatment group (n = 32) and a control group (n = 34). To assess the degree of pituitary desensitization and restoration in the treatment group, gonadotrophin releasing hormone (GnRH) challenges (100 microg, i.v.) were performed during treatment (day 17), and 2, 4 and 6 days after discontinuation of treatment. At the same time blood samples for oestradiol and triptorelin concentrations were drawn. In the control group a GnRH test was performed on day 2 of the menstrual cycle. Both pituitary desensitization during and pituitary recovery after agonist treatment, expressed as the LH response to exogenous GnRH, appeared to be dose dependent. As the use of reduced dosages still offers a considerable degree of pituitary suppression, studies on dose adjustments in the use of triptorelin, in ovarian stimulation in IVF are warranted.

Absorption of an oxytocin antagonist (antocin) and a vasopressin analogue (dDAVP) through a standardized skin erosion in volunteers.

PURPOSE: Transdermal administration of the peptides [Mpa1, D-Tyr (Ethyl)2, Thr4, Orn8]-oxytocin (antocin) and [Mpa1, D-Arg8]-vasopressin (dDAVP) was studied in healthy volunteers. METHODS: A standardized skin erosion was formed preliminary by suctioning. The peptides were administered in plastic reservoirs through a 5 mm erosion and the absorption was followed for a six-day period with plasma concentration determinations on days 1, 3 and 6 with refilling the reservoirs daily with 15 microns and 10 mM solutions of dDAVP and antocin, respectively. Fourteen healthy non-smoking volunteers divided equally between the sexes, participated in the study. Plasma concentrations were measured using specific radioimmunoassays. Reservoir concentrations and metabolic stability of the peptides were determined using reverse-phase HPLC. RESULTS: Both antocin and dDAVP were absorbed across the skin erosion. The absorption pattern was biphasic with a high initial absorption during days 1 and 2 followed by a lower absorption on days 3 and 6. The absorption on day 1, which was estimated at more than 50% for both peptides during a 24 h period, corresponded to a simultaneous decrease in peptide concentration in the reservoirs. The extent of absorption for antocin on days 3 and 6 was 1/3 to 1/6, respectively, of that observed on day 1. Antocin was minimally degraded in the skin reservoir while dDAVP was intact. However, accumulation of cellular material appeared in the antocin reservoirs. The absorption of antocin was reduced by exposure to intact skin surrounding the skin erosion. No pain was experienced and no scar formation was observed. CONCLUSIONS: The observed biphasic absorption may be a consequence of the mild inflammatory response occurring subsequent to eroding the skin. The standardized skin erosion may provide a route for the short-term delivery of otherwise poorly absorbable peptide and protein drugs.

Pharmacokinetic properties of the tocolytic agent [Mpa1, D-Tyr(Et)2, Thr4, Orn8]-oxytocin (antocin) in healthy volunteers.

OBJECTIVE: The aim of this study was to study the pharmacokinetics of antocin, the tocolytic oxytocin antagonist [Mpa1, D-Tyr2(Et), Thr4, Orn8]-oxytocin. DESIGN: Antocin was injected intravenously as a bolus dose (5 mumol). Blood samples were taken at intervals for 240 minutes. In addition, the binding of 125I-Tyr10-antocin to blood constituents was determined and compared with 125I-AVP and 125I-[Mpa1, D-Arg8]-vasopressin (desmopressin). SUBJECTS: Eight healthy, non-smoking adults, three male and five female. MEASUREMENTS: Antocin was measured using a specific radioimmunoassay after prior extraction of the plasma. Plasma binding was estimated using polyethyleneglycol precipitation. RESULTS: The rate of plasma disappearance of antocin was best fitted by a biexponential curve. The clearance of antocin was 23.5 +/- 7.6 l/h, the volume of distribution was 13.1 +/- 3.8 l and the biological half-life was 39.0 +/- 4.1 minutes. A greater proportion of 125I-Tyr10-antocin bound to plasma proteins (33.5%) and red blood cells (13%) than did 125I-AVP, 125I-desmopressin and unlabelled desmopressin. CONCLUSIONS: The half-life was longer and the clearance of antocin was less than that found in a previous study when a non-specific antiserum was used. This is most likely because of the extended blood sampling time period which revealed the biphasic decay pattern. The higher plasma clearance of antocin compared to oxytocin and desmopressin may be explained by its increased binding to blood constituents rather than by differences in enzymatic degradation of the molecules.

Pituitary responsiveness after administration of a GnRH agonist depot formulation: Decapeptyl CR.

OBJECTIVE: This study was focused on the pattern of LH release from the pituitary during the initial response to high dose GnRH agonist administration. Secondly, the pattern of LH release and the pituitary responsiveness to physiological and pharmacological stimulation during long-term pituitary suppression by a high dose GnRH agonist was studied. In addition, the relation between serum agonist levels and pituitary function and responsiveness was investigated. DESIGN: DTrp6GnRH in microcapsules (Decapeptyl CR) was administered i.m. to 12 women on the third day of the cycle. High-rate blood sampling was carried out during the first 48 hours after the injection. Secondly, high-rate blood sampling for 6 hours and a GnRH challenge were performed before and weekly after administration, from week 4 till week 9. All samples were assayed for LH and FSH. LH patterns were analysed by applying a computerized pulse detection program. In the second or third week an oestradiol benzoate test was performed. Finally, triptorelin levels were measured before and weekly after administration. PATIENTS: Twelve patients, suffering from tubal infertility and recruited from the waiting list for in-vitro fertilization/embryo transfer (IVF/ET) participated in the study. RESULTS: During the first 48-hour period, LH and FSH levels demonstrated a rapid rise to peak values after 4 hours, subsequently declining to nearly normal levels. E2 rose to peak values at 12 hours and returned to the follicular range thereafter. LH pulse patterns showed a rapid increase in pulse intervals leading to a near absence of LH pulses at the end of the 48-hour period. From the fourth till the seventh week after agonist administration, LH pulse patterns showed a markedly increased pulse interval, decreased pulse amplitude, and a severely decreased mean LH level. In the same period, LH responses to GnRH were severely blunted or absent. Restoration of the pre-injection LH pulse pattern and the LH response to GnRH was observed during the eighth and ninth week. Oestradiol benzoate challenges showed an E2 rise to preovulatory levels in response to the injections. However, no changes were observed in LH and FSH concentrations. Triptorelin levels showed a peak within 48 hours and gradual decline towards pretreatment values in week eight. CONCLUSIONS: It is concluded from the study, that after administration of triptorelin depot in the early follicular phase, desensitization of the pituitary starts to develop within 24 hours. Pituitary responsiveness is completely absent in the second week and continues to exist until the eighth week after injection, when the agonist has disappeared from the circulation. These findings suggest profound alterations in GnRH receptor availability and post-receptor pathways, that prevent the pituitary from responding to physiological stimuli.

Pharmacokinetic and pharmacologic properties of antiuterotonic oxytocin analogs in the rat.

The pharmacologic and pharmacokinetic properties were evaluated in a series of antiuterotonic oxytocin analogs, modified at positions 1, 2, 4, 8 and, in one case, position 9 of the oxytocin (OT) molecule. [Mpa1,D-Tyr2(Et),Val4,Orn8,desGly9]-OT, [Mpa1,Tyr2(Et),Val4,Orn8]-OT and [Mpa1,D-Tyr2,Val4,Orn8]-OT displayed similar plasma clearance rates (Clps) using the constant infusion method in rats. Two analogs, [Mpa1,D-Tyr2(Et),Val4,Orn8]-OT and, particularly, [Mpa1,D-Tyr2(Et),Thr4,Orn8]-OT, were cleared at significantly higher rates compared with the others. [Mpa1, D-Tyr2(Et), Val4, Orn8]-OT and [Mpa1, D-Tyr2(Et), Thr4, Orn8, desGly9]-OT were most potent in eliciting a short-term in vivo antiuterotonic effect, whereas the duration of effect was longest for [Mpa1, D-Tyr2, Val4, Orn8]-OT and [Mpa1, D-Tyr2(Et), Thr4, Orn8, desGly9]-OT. The Clp of [Mpa1, D-Tyr2, Val4, Orn8]-OT was similar regardless of the infusion rate. No relationship between antiuterotonic effect and Clp of the five peptides could be demonstrated, and no significant linear correlation between Clp and effect duration was found. The apparent volumes of distribution for the present analogs were 10-fold larger than the blood volume, a finding to be considered when measuring in vivo antagonistic activity. The 24-h urinary excretion ranged from 14.3 to 25.6% of the i.v. dose and was negatively correlated with peptide lipophilicity. It is concluded that, in addition to diverging pharmacologic properties, peptide analogs may differ markedly in kinetic parameters like Clp, volumes of distribution and urinary excretion despite minor molecular modifications.

Evaluation of the degradation of desamino1,D-arginine8-vasopressin by nasal mucosa.

The nasal route is a convenient and simple way to administer peptides to humans. Absorption of the drug, however, requires passage of the substance through the nasal mucosa. This is a possible site of enzymatic degradation of the peptide. It is shown that rabbit nasal mucosa homogenates rapidly degrade the synthetic anti-diuretic hormone analogue desamino1,D-arginine8-vasopressin in vitro. The metabolite formed has been identified as des-(amino,arginine8,glycineamide9)- vasopressin, which is stable under the prevalent in vitro incubation conditions. It is proposed that this process is catalyzed by intracellular post-proline cleavage enzyme. Reversed phase chromatography in combination with immunological detection has been used to study the possible presence of this metabolite in the circulation after intranasal administration to humans. The metabolite des-(amino,arginine8,glycineamide9)-vasopressin could not be detected in plasma following intranasal administration, possibly indicating a paracellular absorption of desamino1,D-arginine8-vasopressin or absence of this enzymatic activity in humans.

Differences in transport rate of oxytocin and vasopressin analogues across proximal and distal isolated segments of the small intestine of the rat.

The transmural intestinal passage of some oxytocin and vasopressin analogues (oxytocin, OT; [Mpa1, D-Arg8]vasopressin, dDAVP; [Mpa1, Tyr (OMe)2, carba6]oxytocin, carbetocin; [Mpa1, D-Tyr (OEt)2, Thr4, Orn8]vasotocin, antocin II; [Mpa1, D-Tyr (OEt)2, Thr4, desPro7Orn8Gly9NH2]tocinoic acid-NH(CH2)3NH2, desPOG-antocin II-NH(CH2)3NH2) was studied using isolated proximal and distal segments in the rat. All peptides (measured as peptide-like immunoreactivity) displayed a higher transport rate across distal intestinal segments as determined by radioimmunoassay (RIA). The smallest peptide, des POG-antocin II-NH(CH2)3NH2, was transported at the fastest rate. No correlation of lipophilicity with transport rate was observed. Determination of the amount of peptide remaining in the mucosal media at the end of the incubation period by HPLC did not reveal any visible degradation products. However, the large difference in transport rate between [3H]OT and immunoreactive OT indicates mucosal metabolism of this peptide. [3H]d-DAVP was distributed in a larger mucosal volume than the extracellular space marker [3H]inulin, indicating tissue uptake, but was too low (less than 100% of buffer concentration) to make an active transport mechanism likely. The differences in peptide transport rates between proximal and distal intestinal segments are most likely due to a higher distal paracellular permeability despite a decreased absorptive surface area at this region.