Safety of PEGylated recombinant human full-length coagulation factor VIII (BAX 855) in the overall context of PEG and PEG conjugates.

INTRODUCTION: BAX 855 is a PEGylated human full-length recombinant factor VIII (rFVIII) based on licensed rFVIII (ADVATE). The applied PEGylation technology has been optimized to retain functionality of the FVIII molecule, improve its pharmacokinetic properties and allow less frequent injections while maintaining efficacy. AIM: The aim of this study was to confirm that the excellent safety profile of ADVATE remains unchanged after PEGylation. METHODS: Non-clinical safety studies with BAX 855 and its respective unbound polyethylene glycol (PEG) were conducted in several species. The distribution of a single dose of radiolabelled BAX 855 was further investigated in rats. Publically available safety data on PEG alone and PEGylated biomolecules were summarized and reviewed for specific safety findings attributable to PEG or PEGylated biopharmaceuticals. RESULTS: Safety pharmacology studies in rabbits and macaques and repeated dose toxicity studies in rats and macaques identified no safety issues. Results of a distribution study in rats administered radiolabelled BAX 855 showed that radioactivity was completely excreted; urine was the major elimination route. A 28-day study in rats dosed with the unbound PEG constituent (PEG2ru20KCOOH) of BAX 855 showed no adverse or non-adverse effects. Safety data for PEG and PEG-protein conjugates indicate no safety concerns associated with PEG at clinically relevant dose levels. Although vacuolation of certain cell types has been reported in mammals, no such vacuolation was observed with BAX 855 or with the unbound PEG constituent. CONCLUSION: Non-clinical safety evaluation of PEG and BAX 855 identified no safety signals; the compound is now in clinical development for the treatment of patients with haemophilia A.

Rapid on-site detection of explosives on surfaces by ambient pressure laser desorption and direct inlet single photon ionization or chemical ionization mass spectrometry.

Considering current security issues, powerful tools for detection of security-relevant substances such as traces of explosives and drugs/drug precursors related to clandestine laboratories are required. Especially in the field of detection of explosives and improvised explosive devices, several relevant compounds exhibit a very low vapor pressure. Ambient pressure laser desorption is proposed to make these substances available in the gas phase for the detection by adapted mass spectrometers or in the future with ion-mobility spectrometry as well. In contrast to the state-of-the-art thermal desorption approach, by which the sample surface is probed for explosive traces by a wipe pad being transferred to a thermal desorber unit, by the ambient pressure laser desorption approach presented here, the sample is directly shockwave ablated from the surface. The laser-dispersed molecules are sampled by a heated sniffing capillary located in the vicinity of the ablation spot into the mass analyzer. This approach has the advantage that the target molecules are dispersed more gently than in a thermal desorber unit where the analyte molecules may be decomposed by the thermal intake. In the technical realization, the sampling capillary as well as the laser desorption optics are integrated in the tip of an endoscopic probe or a handheld sampling module. Laboratory as well as field test scenarios were performed, partially in cooperation with the Federal Criminal Police Office (Bundeskriminalamt, BKA, Wiesbaden, Germany), in order to demonstrate the applicability for various explosives, drugs, and drug precursors. In this work, we concentrate on the detection of explosives. A wide range of samples and matrices have been investigated successfully.

[Current strategies in radiotherapy of head- and neck cancer]. / Aktuelle Fortschritte in der Strahlentherapie von Kopf-Hals-Tumoren.

Over the last 20 years there was a dramatic change in therapeutic options for head and neck tumors mainly due to improvements in surgical and radiotherapeutic techniques and in the increasing use of multimodal therapy. Especially for locally advanced tumors, one can achieve long lasting disease free intervals in a large proportion of patients. The following article will focus on the novel techniques in radiation oncology and combined radiochemotherapy giving an overview without too many technical details.

Application of two-dimensional gas chromatography combined with pixel-based chemometric processing for the chemical profiling of illicit drug samples.

The chemical profiling of illicit drugs is a complex process. The results are affected by many different factors such as sample size, the sample processing conditions, the used analytical technique as well as the statistics that are applied. Within this proof-of-concept study, which was done in cooperation with the German Federal Criminal Police Office (Bundeskriminalamt, BKA), the adaptability of comprehensive two-dimensional (2D) gas chromatography (GCxGC) combined with a pixel-based chemometric data processing method is demonstrated. Samples of heroin and cannabis are extracted and analyzed with GCxGC-TOF-MS (time-of-flight mass spectrometry) and GCxGC-FID (flame ionization detection). The obtained second-order data are then used to identify possible marker compounds for the discrimination of the samples according to their chemical profile. The pixel-based chemometric process includes preprocessing steps (background correction, alignment of chromatograms and normalization) followed by an adaptation of hierarchical clustering to identify chemically similar samples, and finally a subsequent calculation of Fisher criterion based on the found clustering in order to identify promising marker compounds. The results of the pixel-based data analysis are compared to a limited peak-based study for cannabis and to a well-established standard method for the chemical profiling of heroin.

Tools for the analysis of dose optimization: III. Pointwise sensitivity and perturbation analysis.

The major challenge in intensity-modulated radiotherapy planning is to find the right balance between tumor control and normal tissue sparing. The most desirable solution is never physically feasible, and a compromise has to be found. One possible way to approach this problem is constrained optimization. In this context, it is worthwhile to quantitatively predict the impact of adjustments of the constraints on the optimum dose distribution. This has been dealt with in regard to cost functions in a previous paper. The aim of the present paper is to introduce spatial resolution to this formalism. Our method reveals the active constraints in a target subvolume that was previously selected by the practitioner for its insufficient dose. This is useful if a multitude of constraints can be the cause of a cold spot. The response of the optimal dose distribution to an adjustment of constraints (perturbation) is predicted. We conclude with a clinical example.

Determination of the ionization potentials of security-relevant substances with single photon ionization mass spectrometry using synchrotron radiation.

Several ionization potentials (IPs) of security relevant substances were determined with single photon ionization time of flight mass spectrometry (SPI-TOFMS) using monochromatized synchrotron radiation from the "Berliner Elektronenspeicherring-Gesellschaft für Synchrotronstrahlung" (BESSY). In detail, the IPs of nine explosives and related compounds, seven narcotics and narcotics precursors, and one chemical warfare agent (CWA) precursor were determined, whereas six IPs already known from the literature were verified correctly. From seven other substances, including one CWA precursor, the IP could not be determined as the molecule ion peak could not be detected. For these substances the appearance energy (AE) of a main fragment was determined. The analyzed security-relevant substances showed IPs significantly below the IPs of common matrix compounds such as nitrogen and oxygen. Therefore, it is possible to find photon energies in between, whereby the molecules of interest can be detected with SPI in very low concentrations due to the shielding of the matrix. All determined IPs except the one of the explosive EGDN were below 10.5 eV. Hence, laser-generated 118 nm photons can be applied for detecting almost all security-relevant substances by, e.g., SPI-TOFMS.

Profiling of new psychoactive substances by using stable isotope ratio mass spectrometry: Study of the synthetic cannabinoid 5F-PB-22.

In this paper, the results of a pilot study on the profiling of the synthetic cannabinoid receptor agonist 5F-PB-22 (5F-QUPIC, pentylfluoro-1H-indole-3-carboxylic acid-8-quinolinyl ester) via isotope ratio mass spectrometry are presented. It focuses on δ13 C, δ15 N and δ2 H isotope ratios, which are determined using elemental analyser (EA) and high temperature elemental analyser (TC/EA) coupled to an isotope ratio mass spectrometer (IRMS). By means of a sample of pure material of 5F-PB-22, it is shown that the extraction of 5F-PB-22 from herbal material, a rapid clean-up procedure, or preparative column chromatography had no influence on the isotope ratios. Furthermore, 5F-PB-22 was extracted from 14 different herbal blend samples ("Spice products" from police seizures) and analysed via IRMS, yielding 3 clusters containing 7, 5, and 2 samples, distinguishable through their isotopic composition, respectively. It is assumed that herbal blends in each cluster have been manufactured from individual batches of 5F-PB-22.

Self-similar chain conformations in polymer gels

We use molecular dynamics simulations to study the swelling of randomly end-cross-linked polymer networks in good solvent conditions. We find that the equilibrium degree of swelling saturates at Q(eq) approximately N(3/5)(e) for mean strand lengths &Nmacr;(s) exceeding the melt entanglement length N(e). The internal structure of the network strands in the swollen state is characterized by a new exponent nu = 0.72+/-0.02. Our findings can be rationalized by a Flory argument for a self-similar structure of mutually interpenetrating network strands, agree partially with the classical Flory-Rehner theory, and are in contradiction to de Gennes' c(*)-theorem.

Tibolone versus conjugated estrogens and sequential progestogen in the treatment of climacteric complaints.

OBJECTIVE: Tibolone has been shown to alleviate climacteric symptoms. This study was designed to compare the effect of tibolone (Livial, 2.5 mg daily) on different climacteric complaints and its impact on the endometrium, determined by vaginal ultrasound, with that of conjugated estrogens (Premarin, 0.625 mg daily) continuously for 6 months in combination with the progestogen medrogestone (Colpron, 2 x 5 mg daily for 12 days each month). METHODS: One hundred and twenty-nine postmenopausal women were recruited and the severity of climacteric symptoms as well as endometrial thickness were recorded at the pre-trial examination and after 1, 3, and 6 months. RESULTS: With the exception of vertigo, mood depression, mood disorder, loss of libido, and dryness of skin, where tibolone was found to be more effective than conjugated estrogens/medrogestone, climacteric symptoms improved significantly in both groups over the 6-month study period. Endometrial thickness did not increase significantly in the tibolone group, whereas in the conjugated estrogens/medrogestone group there was a highly significant increase after 1 month and still a trend towards significance after 6 months. Recurrence of vaginal bleeding occurred significantly less frequently in the tibolone group than in the comparison group. CONCLUSION: Tibolone seems to offer a complete treatment of the climacteric complaints whilst avoiding some of the problems associated with classical hormone replacement therapy.

Ovarian function during low-dose oral contraceptive use.

Lowering the total steroid dose in modern oral contraceptives (OCs) has been connected with a higher incidence of ovarian follicle and cyst formation. To investigate the presence of ovarian follicles and cysts by means of vaginal ultrasonography and serum hormone determinations during use of two low-dose OCs, 65 volunteers were randomized to receive either 20 micrograms ethinylestradiol (EE) + 150 micrograms desogestrel (group A) or 35 micrograms EE + 250 micrograms norgestimate (group B) for a 2-month study period. At baseline, 39% of women in group A and 31% in group B exhibited at least one follicle < 35 mm in diameter. By the end of the second treatment cycle, the frequency of these follicles had decreased to 14% in each group. Only one subject in the higher estrogen group developed an ovarian cyst > 35 mm. One subject in each group demonstrated hormone levels characteristic of ovulation; no pregnancy occurred in either group. The 20 micrograms EE preparation was not found to lead more often to ovarian follicles or cysts when compared with a 35 micrograms EE preparation, possibly because of the type and dose of the progestogen used.

PIP: In Austria, health workers randomly allocated 28 women to the group using the low-dose oral contraceptive (OC) Mercilon (20 mcg ethinyl estradiol [EE] + 150 mcg desogestrel) and 35 women to the group using the low-dose OC Cilest (35 mcg EE + 250 mcg norgestimate). No one had used OCs for at least one month before the study. Clinicians used vaginal ultrasonography and serum hormone levels to learn the degree of ovarian suppression during use of these two low-dose OCs by looking for ovarian follicles and cysts. Before beginning to use the OCs, 39% of women in the Mercilon group and 31% of those in the Cilest group had at least one ovarian follicle. By the second treatment cycle, the frequency of ovarian follicles (35 mm) had fallen significantly to 14% in both groups as compared to baseline (p 0.05). No one in the Mercilon group developed a follicle larger than 35 mm in diameter that remained for more than 4 weeks (i.e., ovarian cyst). One woman in the Cilest group did develop an ovarian cyst (46 mm), however. It appeared during the pill-free week after the first pill cycle and steadily decreased to 40 mm during the second pill cycle. One woman in each group had hormone levels indicative of ovulation. No one in either group became pregnant. These findings suggest that the type and dose of progestogen in the Mercilon OC (desogestrel) were responsible for the lower frequency of ovarian follicles and cysts in the lower-dose OC group than that in the higher-dose OC.

[Ovarian hyperstimulation syndrome]. / Ovarielles Hyperstimulationssyndrom.

The ovarian hyperstimulation syndrome (OHSS) is caused iatrogenically by the application of agents for ovulation induction. The syndrome may be characterized by different symptoms sometimes including an increase in blood viscosity, changes in coagulation parameters as well as renal perfusion. Although the incidence of the severe forms of the syndrome is very low, there is a general increase due to the wide spread use of ovulation induction. The only treatment of the syndrome is prevention by close monitoring of serum oestradiol levels and follicular development.

[Effects of measles vaccination in HIV infected children]. / Skutecznosc szczepienia przeciw odrze u dzieci zakazonych HIV.

OBJECTIVE: The aim of the study was to evaluate the effects of measles vaccination in HIV infected children. METHODS: 13 measles seronegative HIV-infected children received polyvalent measles-mumps-rubella vaccine. MAIN OBSERVATIONS AND RESULTS: Measles seroconversion occurred in 10/13 patients (in revaccinated children in 7/10), rubella seroconversion in 7 out of 9 patients. None of the 12 children responded mumps component. CONCLUSIONS: Measles vaccination seems to be safe and effective in HIV-infected children without severe immunodeficiency.

[Vertical HIV transmission in Poland from 1989-2000]. / Odmatczyne zakazenia HIV w Polsce w latach 1989-2000.

OBJECTIVE: To evaluate vertical HIV transmission rate in Poland. METHODS: 132 children born to HIV positive mothers were evaluated and their HIV infection status was established. MAIN OBSERVATIONS AND RESULTS: There are 60 HIV-infected children in Poland. 92% of them have been infected perinataly. From 1989 to 1994 the rate of vertical transmission was 25%. Since September 1994, when prophylactic strategies were started, the rate has decreased to 21.4% (in children with prophylaxis to 1.8%). CONCLUSIONS: Vertical HIV infection prophylactic programs rely on women's knowledge about their HIV infection status and are the only way to diminish pediatric HIV infection worldwide. The authors show difficulty of providing proper medical care of HIV positive women in Poland and underly the need of voluntary HIV testing for all women before pregnancy.

Health-related quality of life in pediatric patients after allogeneic SCT: development of the PedsQL Stem Cell Transplant module and results of a pilot study.

With increased survival after pediatric allogeneic hematopoietic SCT health-related quality of life (HRQL) has emerged as an essential health outcome. The impact of transplant and chronic GVHD (cGVHD)-associated morbidity remains a major obstacle. In 2005, the National Institutes of Health (NIH) Consensus Conference on Criteria for Clinical Trials in cGVHD recommended HRQL tools as an independent measure of the impact of disease burden. The NIH recommendations did not provide a cGVHD-specific tool for HRQOL measures in children. This report focuses on the development of an SCT-specific instrument to assess HRQL in children and adolescents. For the assessment of generic HRQL we chose the PedsQL (Pediatric Quality of Life Inventory) Generic Cores Scales, which have been used in a large number of healthy, acutely ill and chronically ill children and adolescents. To capture SCT- and, specifically, cGVHD-related problems, we developed the PedsQL Stem Cell Transplant module by reviewing the literature, taking over some items/scales of other PedsQL modules, interviewing patients, parents and members of the health-care team, and applying the PedsQL measurement methods. The final PedsQL Stem Cell Transplant module consists of the HRQL domains: pain and hurt, fatigue/sleeping problems/weakness, nausea, worry/anxiety about disease/treatment, nutritional problems, neurocognitive problems, communication about disease/treatment, loneliness, physical functioning and additional somatic complaints (pruritus, skin inflammation, oral problems, eyes or breathing) including patients' and parents' assessment. It was tested in 35 pediatric patients, who were referred to our SCT Outpatient Clinic about 100 days post SCT. Both the generic PedsQL and the SCT-specific scales showed high internal consistency, with Cronbach alpha levels of ⩾0.70 in almost all scales. Most problems were detected within the HRQL domains of physical functioning and pain. The summary scores of the generic PedsQL and the PedsQL Stem Cell Transplant module showed high correlations (r=0.89 in patients' and r=0.81 in parents' assessments). Moreover, both tools discriminated between patients with and without cGVHD. The PedsQL Stem Cell Transplant module is practical for use and suitable across a broad age range (2-18 years) both in patients with and without cGVHD. However, it is still a pilot instrument and needs further development and testing in a larger patient population.

Forensic profiling of sassafras oils based on comprehensive two-dimensional gas chromatography.

Safrole, the main compound in the essential oil of several plants of the Laurel family (Lauraceae), and its secondary product piperonylmethylketone are the predominantly used precursors for the illicit synthesis of 3,4-methylenedioxymethamphetamine (MDMA) which is, in turn, the most common active ingredient in Ecstasy tablets. Analytical methods with adequate capacity to identify links and origin of precursors, such as safrole, provide valuable information for drug-related police intelligence. Authentic sassafras oil samples from police seizures were subjected to comparative analysis based on their chemical profiles obtained by comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOFMS). The enhanced separation power and increased sensitivity of GC × GC allowed for the detection of minor compounds present in the essential oils which were of particular interest in case of very pure samples whose impurity profiles were not very pronounced. Discrimination of such samples was still possible even in the absence of characteristic main compounds.

Impact of reaction parameters on the chemical profile of 3,4-methylenedioxymethamphetamine synthesized via reductive amination: target analysis based on GC-qMS compared to non-targeted analysis based on GC×GC-TOF-MS.

The most common clandestine manufacturing procedure for the ecstasy derivative 3,4-methylenedioxymethamphetamine (MDMA), is the reductive amination of piperonylmethylketone (PMK) via platinum(IV) oxide/hydrogen. Deviations of the reaction conditions during the synthesis may result in different chemical profiles of the products. The chemical analysis of these profiles is an important objective for forensic drug intelligence. In this work we studied the impact of a systematic variation of the hydrogenation time, the reaction temperature and the precursor batch on the resulting organic chemical profiles of the MDMA bases and MDMA hydrochlorides. Target analysis was based on a gas chromatography mass spectrometry (GC-MS) method which was harmonized during the European project CHAMP.(2) In addition, samples were analyzed by comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS) and subjected to non-targeted data analysis for a comprehensive analysis of the complete profiles. The reaction temperature, followed by the used precursor batch, revealed the highest impact on the chemical profile. The effect on individual impurity compounds is discussed in detail. With respect to the interpretation of the data, the profiles were compared to the profiles of MDMA samples obtained by reductive amination using sodium borohydride ("cold method") and aluminium/mercury amalgam as alternative reducing agents. Non-targeted analysis revealed that the discrimination according to the synthetic route and the batch of precursor used for the synthesis strongly depends on the selected target compounds.

Current strategies in radiotherapy of head and neck cancer.

The Intensity Modulated Radiation Therapy (IMRT) together with Image Guided Radiation Therapy (IGRT) improves radiation therapy for head and neck cancer. On the one hand tumors can be better covered with radiation dose and on the other hand normal tissue can be better preserved. Carefully applied this leads to the same tumor control rate with reduced toxicity compared with conventional radiation therapy. Respective to the addition of systemic therapy to irradiation therapy, platinum based radiochemotherapy remains the standard of care and there are first indications, that at least in the primary treatment of head and neck cancer the addition of Cetuximab to a cisplatin-based radiochemotherapy did not improve outcome.