Whole blood long-chain n-3 fatty acids as a measure of fish oil compliance in children with acute lymphoblastic leukemia: a pilot study.

Long-chain n-3 fatty acids (n-3 LCPUFA) may prevent chemotherapy-induced hyperlipidemia in children with acute lymphoblastic leukemia (ALL). However, compliance could be a problem and intake-biomarker correlations may be affected by bodyweight and blood transfusions. We assessed whole blood n-3 LCPUFA three times during the first 83 days of treatment in six 1-17-year-old children with ALL, who received 2.4-4.9 g/d n-3 LCPUFA depending on bodyweight. Mean compliance was 73%, which resulted in a 2.5-fold increase in blood n-3 LCPUFA irrespective of blood transfusions. The correlation between relative blood content of n-3 LCPUFA and intake in g/d across the study period was strong (r=0.76, p=0.001). When n-3 LCPUFA was expressed in absolute concentrations and intake per kg bodyweight the correlation decreased (r=0.39, p=0.164) and was driven by baseline values. Thus, relative content of n-3 LCPUFA in blood reflects fish oil compliance in children with ALL despite blood transfusions and differences in bodyweight.

Incidence trends of melanoma of the skin compared with other localisations, in the Norwegian population, 1956-2005.

BACKGROUND: Increasing incidence rates (IRs) of cutaneous malignant melanoma (CMM) have been associated with increased exposure to ultraviolet radiation (UVR). The aim of this study was to compare the changes over time in IRs of malignant melanoma in anatomical localisations exposed to different levels of UVR in the same population. PATIENTS AND METHODS: All incident cases of melanoma (invasive) diagnosed 1956-2005 were extracted from the database of the Cancer Registry of Norway. The average percentage change of the age-standardised IRs per 5-year period of diagnosis was calculated (loglinear regression) by anatomical localisation (skin, eye, internal organs and external genitalia). RESULTS: CMM represented 91.7% of the total number of melanomas, while ocular melanoma and melanoma in the internal organs represented 6.2% and 1.2%, respectively. The average quinquennal percentage increase in IRs for CMM and melanoma in internal organs was 23.3% [95% confidence interval (CI) 20.9-25.8] and 14.0% (95% CI 8.2-19.7), respectively. The corresponding analysis for ocular melanoma showed an increase of 1.3% (95% CI -1.5 to 4.2) and a decrease (not significant) for melanoma on male (-8.8%) and female (-2.1%) external genitalia. CONCLUSION: A wide variation in IRs and trends between the four anatomical localisations with unlike levels of UVR exposure suggests different causal pathways for melanoma.

Gravity amplifies and microgravity decreases circumnutations in Arabidopsis thaliana stems: results from a space experiment.

In a microgravity experiment onboard the International Space Station, circumnutations of Arabidopsis thaliana were studied. Plants were cultivated on rotors under a light:dark (LD) cycle of 16 : 8 h, and it was possible to apply controlled centrifugation pulses. Time-lapse images of inflorescence stems (primary, primary axillary and lateral inflorescences) documented the effect of microgravity on the circumnutations. Self-sustained circumnutations of side stems were present in microgravity but amplitudes were mostly very small. In darkness, centrifugation at 0.8 g increased the amplitude by a factor of five to ten. The period at 0.8 g was c. 85 min, in microgravity roughly of the same magnitude. In white light the period decreased to c. 60 min at 0.8 g (microgravity value not measurable). Three-dimensional data showed that under 0.8 g side stems rotated in both clockwise and counter-clockwise directions. Circumnutation data for the main stem in light showed a doubling of the amplitude and a longer period at 0.8 g than in microgravity (c. 80 vs 60 min). For the first time, the importance of gravity in amplifying minute oscillatory movements in microgravity into high-amplitude circumnutations was unequivocally demonstrated. The importance of these findings for the modelling of gravity effects on self-sustained oscillatory movements is discussed.

Ultradian rhythms in Arabidopsis thaliana leaves in microgravity.

Ultradian movements of Arabidopsis thaliana rosette leaves were discovered and studied under microgravity conditions in space. Weightlessness revealed new facets of these movements. The European Modular Cultivation System (EMCS) was used in a long-term white-light, light-darkness (LD; 16 : 8 h) experiment on the International Space Station (ISS). Leaves reacted with slow up or down movement (time constant several hours) after transitions to darkness or light, respectively. Superimposed movements with periods of c. 80-90 min and small-amplitude pulsed movements of 45 min were present in the light. Signal analysis (fast Fourier transform (FFT) analysis) revealed several types and frequencies of movements. Identical phase coupling was observed between the 45-min movements of the leaves of one plant. In darkness, movements of c. 120-min period were recorded. The EMCS allowed 0-g to 1-g transitions to be created. Leaves on plants germinated in microgravity started a negative gravitropic reaction after a delay of c. 30 min. Leaves grown on a 1-g centrifuge reacted to the same transition with an equal delay but had a weaker gravitropic response. The experiments provide unequivocal demonstrations of ultradian, self-sustained rhythmic movements in A. thaliana rosette leaves in the absence of the effect of gravity.

A PDRMIP multi-model study on the impacts of regional aerosol forcings on global and regional precipitation.

Atmospheric aerosols such as sulfate and black carbon (BC) generate inhomogeneous radiative forcing and can affect precipitation in distinct ways compared to greenhouse gases (GHGs). Their regional effects on the atmospheric energy budget and circulation can be important for understanding and predicting global and regional precipitation changes, which act on top of the background GHG-induced hydrological changes. Under the framework of the Precipitation Driver Response Model Inter-comparison Project (PDRMIP), multiple models were used for the first time to simulate the influence of regional (Asian and European) sulfate and BC forcing on global and regional precipitation. The results show that, as in the case of global aerosol forcing, the global fast precipitation response to regional aerosol forcing scales with global atmospheric absorption, and the slow precipitation response scales with global surface temperature response. Asian sulphate aerosols appear to be a stronger driver of global temperature and precipitation change compared to European aerosols, but when the responses are normalised by unit radiative forcing or by aerosol burden change, the picture reverses, with European aerosols being more efficient in driving global change. The global apparent hydrological sensitivities of these regional forcing experiments are again consistent with those for corresponding global aerosol forcings found in the literature. However, the regional responses and regional apparent hydrological sensitivities do not align with the corresponding global values. Through a holistic approach involving analysis of the energy budget combined with exploring changes in atmospheric dynamics, we provide a framework for explaining the global and regional precipitation responses to regional aerosol forcing.

Endosome to Golgi transport of ricin is regulated by cholesterol.

We have here studied the role of cholesterol in transport of ricin from endosomes to the Golgi apparatus. Ricin is endocytosed even when cells are depleted for cholesterol by using methyl-beta-cyclodextrin (m beta CD). However, as here shown, the intracellular transport of ricin from endosomes to the Golgi apparatus, measured by quantifying sulfation of a modified ricin molecule, is strongly inhibited when the cholesterol content of the cell is reduced. On the other hand, increasing the level of cholesterol by treating cells with mbetaCD saturated with cholesterol (m beta CD/chol) reduced the intracellular transport of ricin to the Golgi apparatus even more strongly. The intracellular transport routes affected include both Rab9-independent and Rab9-dependent pathways to the Golgi apparatus, since both sulfation of ricin after induced expression of mutant Rab9 (mRab9) to inhibit late endosome to Golgi transport and sulfation of a modified mannose 6-phosphate receptor (M6PR) were inhibited after removal or addition of cholesterol. Furthermore, the structure of the Golgi apparatus was affected by increased levels of cholesterol, as visualized by pronounced vesiculation and formation of smaller stacks. Thus, our results indicate that transport of ricin from endosomes to the Golgi apparatus is influenced by the cholesterol content of the cell.

Endosome to Golgi transport of ricin is independent of clathrin and of the Rab9- and Rab11-GTPases.

The plant toxin ricin is transported to the Golgi and the endoplasmic reticulum before translocation to the cytosol where it inhibits protein synthesis. The toxin can therefore be used to investigate pathways leading to the Golgi apparatus. Except for the Rab9-mediated transport of mannose 6-phosphate receptors from endosomes to the trans-Golgi network (TGN), transport routes between endosomes and the Golgi apparatus are still poorly characterized. To investigate endosome to Golgi transport, we have used here a modified ricin molecule containing a tyrosine sulfation site and quantified incorporation of radioactive sulfate, a TGN modification. A tetracycline-inducible mutant Rab9S21N HeLa cell line was constructed and characterized to study whether Rab9 was involved in transport of ricin to the TGN and, if not, to further investigate the route used by ricin. Induced expression of Rab9S21N inhibited Golgi transport of mannose 6-phosphate receptors but did not affect the sulfation of ricin, suggesting that ricin is transported to the TGN via a Rab9-independent pathway. Moreover, because Rab11 is present in the endosomal recycling compartment and the TGN, studies of transient transfections with mutant Rab11 were performed. The results indicated that routing of ricin from endosomes to the TGN occurs by a Rab11-independent pathway. Finally, because clathrin has been implicated in early endosome to TGN transport, ricin transport was investigated in cells with inducible expression of antisense to clathrin heavy chain. Importantly, endosome to TGN transport (sulfation of endocytosed ricin) was unchanged when clathrin function was abolished. In conclusion, ricin is transported from endosomes to the Golgi apparatus by a Rab9-, Rab11-, and clathrin-independent pathway.

PDRMIP: A Precipitation Driver and Response Model Intercomparison Project, Protocol and preliminary results.

As the global temperature increases with changing climate, precipitation rates and patterns are affected through a wide range of physical mechanisms. The globally averaged intensity of extreme precipitation also changes more rapidly than the globally averaged precipitation rate. While some aspects of the regional variation in precipitation predicted by climate models appear robust, there is still a large degree of inter-model differences unaccounted for. Individual drivers of climate change initially alter the energy budget of the atmosphere leading to distinct rapid adjustments involving changes in precipitation. Differences in how these rapid adjustment processes manifest themselves within models are likely to explain a large fraction of the present model spread and needs better quantifications to improve precipitation predictions. Here, we introduce the Precipitation Driver and Response Model Intercomparison Project (PDRMIP), where a set of idealized experiments designed to understand the role of different climate forcing mechanisms were performed by a large set of climate models. PDRMIP focuses on understanding how precipitation changes relating to rapid adjustments and slower responses to climate forcings are represented across models. Initial results show that rapid adjustments account for large regional differences in hydrological sensitivity across multiple drivers. The PDRMIP results are expected to dramatically improve our understanding of the causes of the present diversity in future climate projections.

Specific pathogen-free pig herds also free from Campylobacter?

As Specific Pathogen-Free (SPF) pig herds are designed and managed to prevent specific pig diseases, it might be feasible to expand the list of micro-organisms also including zoonotic pathogens such as Campylobacter coli as this agent has its origin in pigs. In a previous survey, 15 of 16 of SPF herds were found free from human pathogenic Yersinia enterocolitica. Accordingly, three nucleus and seven multiplying herds were surveyed for Campylobacter to investigate whether the Norwegian SPF pig pyramid also might be free from this agent. In conclusion, the intervention of Campylobacter at the herd level might be possible as four of 10 SPF herds tested negative in two sets of samples from both autumn 2008 and summer/early autumn 2010. The four negative herds were all located in remote areas several kilometres away from conventional pig farming while the positive SPF farms were all situated in neighbourhoods with conventional pig production. It seems more difficult to control Campylobacter than some specific animal disease agents and another significant zoonotic agent, Y. enterocolitica, in pig herds.

Intraepithelial and invasive squamous cell neoplasia of the vulva: trends in incidence, recurrence, and survival rate in Norway.

OBJECTIVE: To characterize time trends in incidence of intraepithelial and invasive vulvar neoplasia, transition of intraepithelial vulvar neoplasia to invasive cancer, and survival rate based on a total population. METHODS: The Cancer Registry of Norway was used to identify all Norwegian inhabitants diagnosed during 1956-1990 with squamous cell carcinoma of the vulva, and all those with intraepithelial vulvar neoplasia diagnosed during 1973-92. RESULTS: The incidence rate of intraepithelial neoplasia increased three-fold from 1973-77 to 1988-92, and 3.4% changed into invasive disease. Multivariate analyses identified age as a significant variable. The age-adjusted incidence rate for squamous cell carcinoma was constant for the whole period. The ratio between recurrence and the total number of invasive cancer varied from 21% to 31%. The 5-year survival rates have not changed significantly over the period. Age was revealed as a strong prognostic factor, showing an excess death rate by increasing age. CONCLUSION: The incidence of vulvar intraepithelial neoplasia has increased substantially over the last 40 years, but that of invasive vulvar cancer has not changed appreciably.

Individualized treatment of stage I carcinoma of the vulva.

A series of 117 patients with stage I squamous cell carcinoma of the vulva was followed for 3 to 21 years. Twenty recurrences (17.1%) were found, 12 in the vulva or vagina, 7 in the groin, and 1 in a patient who developed distant metastases. Invasion of tumor cells in lymph or blood vessels was found in operative specimens from 19 patients (16.2%), 8 of whom (42.1%) developed local recurrences of metastases. Of 76 patients who underwent lymphadenectomy, 7 had ipsilateral and 1 had bilateral metastases to the inguinal lymph nodes. Five-year crude survival rate for the whole series was 79%, but only 52% for the group of patients with vessel invasion. Treatment should in most cases be hemivulvectomy with ipsilateral inguinal lymphadenectomy.

Stage IB carcinoma of the cervix, the Norwegian Radium hospital. II. Results when pelvic nodes are involved.

From 1967 through 1972, 562 patients with squamous cell carcinoma of the cervix, stage IB (FIGO), underwent radical hysterectomy with pelvic lymphadenectomy. The patients with pelvic metastases were treated with external beam supervoltage irradiation of 4000 or 5000 rads to the pelvis. Metastases in the pelvic lymph nodes were found in 21.3%. In this study the authors analyzed in detail which nodes were involved with metastases and the relationship with survival and with recurrence. The overall 5-year survival for patients with pelvic lymph node metastases was 53%; for patients without metastases it was 92%. The site of recurrence in patients without demonstrable pelvic lymph node metastases was also examined. The main conclusion drawn from the study is that patients with pelvic node metastases below the common iliac group have a much better prognosis than patients in whom metastases are located in the common iliac nodes; in addition, when nodal involvement is below the common iliac level, patients with disease limited to one node or one group of nodes do not have a better prognosis than patients with multiple metastatic nodes.

Single and repeated dose pharmacokinetics of thio-TEPA in patients treated for ovarian carcinoma.

Triethylenethiophosphoramide (thio-TEPA) pharmacokinetics were studied in 15 patients being treated for epithelial ovarian carcinoma. Unchanged thio-TEPA was assayed in serum and urine by means of a gas chromatographic procedure. No accumulation or alteration of the pharmacokinetics occurred during therapy, which was continued for up to 7 months with biweekly administrations of 20 mg, after two initial loading courses with 20 mg daily for 3 consecutive days 2 weeks apart. No significant difference in the pharmacokinetics between i.m. and i.v. administration was demonstrated. However, three patients showed a reduced absorption ability from the i.m. injection site to the systemic circulation and an apparent increase in the elimination half-life (3.86 +/- 0.97 h), which could be of clinical relevance. A first-order elimination process with a short elimination half-life (approximately 1.5 h) was demonstrated for thio-TEPA in all patients after i.v. administration. The apparent volume of distribution averaged 50 1. The renal clearance was below 1% of the total-body clearance, which averaged 412 ml/min. The urinary excretion of unchanged thio-TEPA was complete within 8 h after administration, with an average urinary recovery of 0.14% of the dose. Calculation of the area under the serum concentration vs time curve revealed wide variation between patients (range 517-1480 ng/h ml-1), indicating the need for drug monitoring during therapy.

The behaviour of normal and agravitropic transgenic roots of rapeseed (Brassica napus L.) under microgravity conditions.

In the TRANSFORM experiment for IML-2 on the Space Shuttle Columbia, normal (wild type = WT) and genetically transformed agravitropic rapeseed roots were tested under microgravity conditions. The aim of the experiment was to determine if the wild-type roots behaved differently (growth, morphology, gravitropical sensitivity) from the transgenic roots. The appearance of the organelles and distribution of statoliths (i.e. amyloplasts with starch grains) in the gravitropic reactive cells (statocytes) under weightlessness was compared for the two types of roots. Attempts have also been made to regenerate new plants from the root material tested in space. Both the WT and the transgenic root types showed the expected increase in length during 36 h of photorecording. Contrary to the results of the ground controls, no significant difference in elongation rates was found between the WT and transgenic roots grown in orbit. However, there are indications that the total growth both in the WT and the transgenic roots was higher in the ground control than for roots in orbit. After a 60 min 1 x g stimulation of the roots on board the Shuttle, no detectable curvatures were obtained in either the transgenic or the WT roots. However, it cannot be excluded that a minute curvature development occurs in the root tips but was not detected due to technical reasons. The ultrastructure was well preserved in both the WT and the transgenic roots, despite the fact that the tissue was kept in the prefixative for over 3 weeks. No marked differences in ultrastructure were observed between the transformed root statocyte cells and the equivalent cells in the wild type. There were no obvious differences in root morphology during the orbital period. Light micrographs and morphometrical analysis indicate that the amyloplasts of both the wild type and transformed root statocytes are randomly distributed over the cells kept under micro-g conditions for 37 h after a 14 h stimulation on the 1 x g centrifuge. The main scientific conclusion from the TRANSFORM experiment is that the difference in growth found in the ground control between the WT and the transgenic root types seems to be eliminated under weightlessness. Explanations for this behaviour cannot be found in the root ultrastructure or in root morphology.

Dynamics of root growth in microgravity.

An experiment to study the growth of garden cress roots in microgravity is described. The experiment, denoted RANDOM, was an ESA Biorack experiment in the IML-2 flight in July 1994. In the absence of gravity, it can be anticipated that the roots would show random growth, changing their direction randomly. The hypothesis that such random growth movements occur according to random walk theory, leads to predictions as to the detailed manner in which deviations increase with time. The experiment was designed to test this random walk hypothesis. The paper concentrates on the technological aspects of studying the roots in microgravity. The development of suitable plant chambers, fitting containers developed by ESA, is described as well as the techniques used to grow the seeds between agar slices. hardware was developed to record photographically root movements between the agar slices. Photos were taken once per hour. Some plant chambers were designed to allow fixation of plant material in space. The practical solutions found using glutaraldehyde for prefixation in the Spacelab, within the restrictions given, are described. The experimental results show that the growth pattern in fact followed the prediction from the random walk approach. The average changes in the growth direction stayed constant and equal to zero during the experiment while the squared angular deviations increased proportional to time. Furthermore, plant material prefixed in orbit was permanently fixed after the flight. Light microscopy and electron microscopy pictures are shown as examples of the results achieved. The long prefixation period meant a drawback for the quality of the fixation process. However, sections suitable for study were achieved. The main goals of the RANDOM experiment were therefore achieved.

The effect of a private sector on the waiting time in national health service.

This article examines the effect of a private sector on the waiting time associated with treatment in a public hospital. Without rationing of waiting-list admissions, a private sector is shown to result in a longer waiting time if the demand for a public treatment is sufficiently elastic with respect to the waiting time. When waiting-list admissions are rationed, the waiting time is shown to increase if the public sector consultants are permitted to work in the private sector in their spare time.

A theory of hospital waiting lists.

The noncooperative character of resource allocation in a national health service may contribute to excessive waiting lists. A theory of hospital waiting lists is derived from this idea. Waiting lists imply loss of efficiency; the hospital's resources are drawn away from medical work. Although there is scope for Pareto improvements, the structure of budget allocation may prevent these improvements from being achieved. Some reforms of the institutional structure are suggested.

Flow cytometric DNA measurements in squamous cell carcinoma of the vulva: an important prognostic method.

One hundred and thirty-one squamous cell carcinomas of the vulva were examined by FCM-DNA measurements. Samples were prepared from paraffin-embedded tissue. Of these, 66 were found to be diploid, 52 aneuploid and 13 could not be evaluated. The 5-year crude survival rate was 62% for the diploid and 23% for the aneuploid tumors (P < 0.001). The aneuploid tumors without lymph node (LN) metastases showed a 5-year cancer-related survival rate of 44% as compared to 58% for the diploid tumors with LN metastases. In a multivariate Cox regression analysis the most important independent prognostic parameters were (1) LN involvement (P < 0.0001), (2) tumor ploidy (P = 0.0001) and (3) tumor size (P = 0.0039). By using ploidy and lymph node involvement in this way as prognostic factors we are able to identify high- and low-risk groups of patients. We strongly believe that these results should lead to a different attitude towards therapy in vulva cancer patients.

Antigenic determinants of Salmonella serogroups A and D1. Synthesis of trisaccharide glycosides for use as artificial antigens.

The disaccharide glycoside 8-methoxycarbonyloctyl 4,6-O-cyclohexylidine-2-O-(tetra-O-benzyl-alpha-D-galactopyranosyl)-alpha-D-manno pyranoside (7) was used as a common intermediate to the trisaccharide determinants of both Salmonella serogroups A and D1. Acetalation of 8-methoxycarbonyloctyl alpha-D-mannopyranoside provided the 4,6-acetal derivative, which was selectively benzoylated to give the partially protected mannoside 4. Reaction of 4 with tetra-O-benzyl-alpha-D-galacto-pyranosyl chloride afforded the fully protected disaccharide, which, after transesterification, gave the selectively blocked, disaccharide glycoside (7). Addition of tyvelose by way of its 2,4-di-O-benzoyl-3,6-dideoxy-alpha-D-arabino-hexopyranosyl chloride derivative gave the blocked trisacchride determinant of Salmonella serogroup D1. 2,4,-Di-O-benzyl-3,6-dideoxy-alpha-D-ribo-hexopyranosyl chloride reacted with 7 to provide, after removal of blocking groups, the paratose-containing determinant of serogroup A.

Synthesis of the trisaccharide moiety of gangliotriosylceramide (asialo GM2).

The synthesis of the trisaccharide methyl glycoside beta-D-GalNAc-(1----4)-beta-D-Gal-(1----4)-beta-D-Glc-OMe, which corresponds to the carbohydrate portion of gangliotriosylceramide (asialo GM2), was accomplished by the reaction of 4-O-acetyl-3,6-di-O-benzoyl-2-deoxy-2-phthalimido-D-galactopyranosyl bromide (18) with a benzylated derivative of methyl 4-O-beta-D-galactopyranosyl-beta-D-glucopyranoside. Comparative studies with a 6'-benzyl ether and a 6'-benzoate revealed that the substituent at O-6' is crucial to the outcome of glycosylations at O-4', the ether derivative being much the more reactive. tert-Butyl 4-O-acetyl-3,6-di-O-benzoyl-2-deoxy-2-phthalimido-D-galactopyranoside, which was readily converted into the corresponding bromide 18, was obtained from the gluco derivative via a single-step, crown ether-assisted epimerization.