CD40-targeted adenoviral gene transfer to dendritic cells through the use of a novel bispecific single-chain Fv antibody enhances cytotoxic T cell activation

Adenoviral (Ad) transduction of dendritic cells (DC) is a promising vaccination strategy. However, clinical applicability of Ad vectors is hampered by the necessity to use high titers of infectious Ad particles for efficient DC transduction. Here, we report on the production of a bacterially expressed bispecific conjugate, consisting of a fusion of recombinant single-chain (sc) mAb Fv fragments, which bind and neutralize the Ad fiber knob (through the S11 mAb scFv) and retarget Ad to CD40 on the DC surface (through the G28-5 mAb scFv). We show that this bispecific scFv fusion protein significantly enhances transduction efficiency of monocyte-derived DC (MoDC), reduces the amount of virus needed for a given level of transduction, and increases the ability of MoDC to activate CTL in an antigen specific manner. This single-component conjugate may prove to be a valuable immunotherapeutic tool for the targeting of Ad to DC in vivo.     

Health Council report on health claims for foods and food supplements

The Health Council of the Netherlands has issued a report on the health claims often made for foods and food supplements. It is concluded that the boundary between medical claims and health claims is not clear: there is no scientific or practical distinction between the 'prevention of disease' and the 'promotion and maintenance of health'. Only explicit, specific and evidence-based claims regarding the prevention, and possibly the treatment, of diseases or conditions are of any value to public health. If the impression is correct that the nutrient content of a product, a change in bodily function or a reduction of a disease risk factor reduces the likelihood of developing a certain disease, this effect should be stated explicitly. If the impression is not correct, then the claim is irrelevant and therefore misleading. A claim must always be supported by research involving human subjects and generally accepted scientific criteria must be adhered to.     

Short and long term variability of the interrupter technique under field and standardised conditions in 3-6 year old children

BACKGROUND: The short and long term variability of the interrupter technique was assessed to determine whether interrupter resistance is a stable individual characteristic over time. The effect of field and standardised measurement conditions on the within-subject variability of the interrupter technique was also examined. METHODS: The interrupter technique was studied under field and standardised conditions in children aged 3-6 years. Under field conditions, five investigators performed the measurements using two different measurement devices in random sequence. Both short term (20-30 minutes) and long term variability (median 38 days) were assessed in 32 children. Under standardised conditions, a single investigator conducted all measurements using a single device; the repeated measurements were conducted at the same time of day in a familiar quiet classroom. Long term variability (median 11 days) was estimated in 15 children. Within-subject standard deviations were estimated by analysis of variance with adjustment for the effects of different investigators and measurement devices on within-subject variability under field conditions. RESULTS: Under field conditions within-subject standard deviations for short and long term variability were 0.10 kPa/l/s (adjusted 0.10 kPa/l/s) and 0.13 kPa/l/s (adjusted 0.14 kPa/l/s), respectively. Under standardised conditions the within-subject standard deviation for long term variability was 0.10 kPa/l/s. CONCLUSIONS: Measurement of interrupter resistance under field conditions only slightly increased the within-subject variability compared with standardised conditions. The results indicate that interrupter resistance is a stable individual characteristic over a period of some weeks.     

Use of health care services by Afghan, Iranian, and Somali refugees and asylum seekers living in The Netherlands

BACKGROUND: Although asylum seekers have been coming to The Netherlands since the 1980s, very few epidemiological studies have focused on this group of inhabitants, or on the refugees who have resettled in this country. The objective of this study is to estimate the use of health care services by refugees and asylum seekers and to identify determinants for this utilisation. METHODS: A population-based study was conducted in The Netherlands from June 2003 to April 2004 among adult refugees and asylum seekers from Afghanistan, Iran, and Somalia. A total of 178 refugees and 232 asylum seekers, living in 3 municipalities and 14 reception centres, participated. RESULTS: This study showed that there are no differences between refugees and asylum seekers in the self-reported use of health care services. Respondents from Somalia reported less contacts with a general practitioner, less use of mental health services, and less medication use than respondents from Afghanistan and Iran. Both female gender and older age were related to more contacts with a general practitioner and a medical specialist, and with higher medication use. Poor general health was related to more contacts with a medical specialist and mental health services, and with higher medication use. CONCLUSION: Asylum seekers and refugees seem to have equal access to the Dutch health care system in general. However, there are differences in the self-reported use of health care services by the different ethnic groups.     

Potentiation of radiation therapy by the oncolytic adenovirus dl1520 (ONYX-015) in human malignant glioma xenografts

In spite of aggressive surgery, irradiation and/or chemotherapy, treatment of malignant gliomas remains a major challenge in adults and children due to high treatment failure. We have demonstrated significant cell lysis and antitumour activity of the E1B-55 kDa-gene-deleted adenovirus ONYX-015 (dl1520, CI-1042; ONYX Pharmaceuticals) in subcutaneous human malignant glioma xenografts deriving from primary tumours. Here, we show the combined efficacy of this oncolytic therapy with radiation therapy. Total body irradiation (5 Gy) of athymic nude mice prior to intratumoral injections of ONYX-015 1 x 10(8) PFU daily for 5 consecutive days yielded additive tumour growth delays in the p53 mutant xenograft IGRG88. Radiation therapy was potentiated in the p53 functional tumour IGRG121 with a 'subtherapeutic' dose of 1 x 10(7) PFU daily for 5 consecutive days, inducing significant tumour growth delay, 90% tumour regression and 50% tumour-free survivors 4 months after treatment. These potentiating effects were not due to increased adenoviral infectivity or replication. Furthermore, cell lysis and induction of apoptosis, the major mechanisms for adenoviral antitumour activity, did not play a major role in the combined treatment strategy. Interestingly, the oncolytic adenovirus seemed to accelerate radiation-induced tumour fibrosis. Potentiating antitumour activity suggests the development of this combined treatment for these highly malignant tumours.     

Morphological and electrophysiological study of the effects of cisplatin and ORG.2766 on rat spinal ganglion neurons

Eleven- to 12-wk-old rats were treated twice a week with cisplatin/saline or with cisplatin plus ORG.2766 during 12.5 wk. Cisplatin and ORG.2766 were administered at a final concentration of 0.04 mg/ml (i.p.) and 10 micrograms/ml (s.c.), respectively. Control animals were treated with saline. In this period the cisplatin-treated animals developed a peripheral neuropathy resulting in impairment of sensory functions. Estimates of the motor (MNCV) and sensory (SNCV) nerve conduction velocity were made after 0, 7.5, 10, and 12.5 wk. It appeared that the MNCV of the control, cisplatin-, and cisplatin plus ORG.2766-treated rats increased from 50 to 59 m/s. In contrast, the SNCV of the cisplatin-treated rats decreased significantly (P less than 0.001) from 63 to 56 m/s, whereas that of the control animals increased from 62 to 84 m/s. Rats which received cisplatin plus ORG.2766 showed an increase in SNCV up to control levels. After 12.5 wk the animals were perfused with a mixture of 1% paraformaldehyde and 1.25% glutaraldehyde in 0.05 M phosphate buffer. At the level of L5 and L6, 5 mm of spinal cord tissue and three dorsal root ganglia were removed and processed for electron microscopy. With the point-counting method the volume fraction (v/v) of somata and myelin in spinal ganglia was estimated. No significant change in the volume fraction of somata of the control (0.42), cisplatin (0.33)-, and cisplatin plus ORG.2766 (0.39)-treated rats was found. The same held true for the volume fraction of myelin of the control (0.53), cisplatin (0.59)-, and cisplatin plus ORG.2766 (0.58)-treated rats. In addition, the number of lysosomes per 100 microns 2 was estimated in spinal ganglion neurons and in spinal cord motor neurons of a total of 120 randomly chosen neurons. It was found that the number of lysosomes in the spinal ganglion neurons of the control animals was lower (10 per 100 microns 2) than in cisplatin-treated (30 per 100 microns 2) and in cisplatin plus ORG.2766-treated rats (28 per 100 microns 2) (P less than 0.05). No difference was observed in the number of lysosomes between cisplatin- and cisplatin plus ORG.2766-treated rats. The number of lysosomes in spinal cord tissue of cisplatin-treated rats (2.4 per 100 microns 2) did not differ from controls (0.1 per 100 microns 2) and from cisplatin plus ORG.2766-treated rats (0.8 per 100 microns 2).(ABSTRACT TRUNCATED AT 400 WORDS)     

Budesonide and terbutaline or terbutaline alone in children with mild asthma: effects on bronchial hyperresponsiveness and diurnal variation in peak flow.

The effects of treatment with budesonide (200 micrograms twice daily) and terbutaline (500 micrograms four times daily) has been compared with the effects of placebo and terbutaline in 27 children with mild asthma, aged 7-14 years, in a double blind, randomised placebo controlled study over eight weeks. Bronchial responsiveness (PC20 histamine), lung function, the amplitude of diurnal variation in peak expiratory flow (PEF), and symptom scores were measured. Baseline FEV1 was over 70% predicted and PC20 histamine less than 8 mg/ml. Twelve children were treated with budesonide and terbutaline and 15 with placebo and terbutaline. After four and eight weeks of treatment the change in PC20 was significantly greater after budesonide and terbutaline than after terbutaline alone by 2.1 (95% CI 0.5-3.8) and 1.3 (95% CI 0.1-2.5) doubling doses respectively. Mean FEV1 did not change in either group. The change in afternoon and nocturnal PEF was significantly greater after budesonide and terbutaline than after terbutaline alone. The amplitude of diurnal variation in PEF did not change significantly in either group. Peak flow reversibility decreased in the budesonide group. There were no differences between treatments for cough and dyspnoea, but wheeze improved in the budesonide group. The children with mild asthma treated with budesonide and terbutaline showed improvement in bronchial responsiveness, afternoon and nocturnal PEF, and symptoms of wheeze and a fall in peak flow reversibility by comparison with those who received terbutaline alone.     

Detection of leptospires in urine by polymerase chain reaction.

Primers for polymerase chain reaction (PCR) were synthesized from clones derived from a Leptospira hardjo (type hardjobovis) library. One pair of synthetic oligonucleotide primers was selected for further analysis. Under experimental conditions an amplification was obtained with DNA of Leptospira interrogans of some serovars belonging to serogroup sejroe. However, very little or no amplification was observed with DNA from other serovars of this group. No amplification was observed with DNA from other serogroups, other bacteria, or eucaryotic organisms. Cattle urine, seeded with hardjobovis, was processed in several ways and subsequently subjected to PCR. Boiling of the samples or treatment with detergents appeared to be most effective. Urine samples containing fewer than 10 leptospires gave a positive result in the PCR assay. Twenty urine samples obtained from a slaughterhouse or farm cows were investigated using the PCR assay, culture isolation, dot and quick blot hybridization, and serological tests. This comparative study suggests that amplification by PCR may be a valuable method for the detection of leptospires in cattle urine.