FDA-approved drug screening in patient-derived organoids demonstrates potential of drug repurposing for rare cystic fibrosis genotypes.

BACKGROUND: Preclinical cell-based assays that recapitulate human disease play an important role in drug repurposing. We previously developed a functional forskolin induced swelling (FIS) assay using patient-derived intestinal organoids (PDIOs), allowing functional characterization of CFTR, the gene mutated in people with cystic fibrosis (pwCF). CFTR function-increasing pharmacotherapies have revolutionized treatment for approximately 85% of people with CF who carry the most prevalent F508del-CFTR mutation, but a large unmet need remains to identify new treatments for all pwCF. METHODS: We used 76 PDIOs not homozygous for F508del-CFTR to test the efficacy of 1400 FDA-approved drugs on improving CFTR function, as measured in FIS assays. The most promising hits were verified in a secondary FIS screen. Based on the results of this secondary screen, we further investigated CFTR elevating function of PDE4 inhibitors and currently existing CFTR modulators. RESULTS: In the primary screen, 30 hits were characterized that elevated CFTR function. In the secondary validation screen, 19 hits were confirmed and categorized in three main drug families: CFTR modulators, PDE4 inhibitors and tyrosine kinase inhibitors. We show that PDE4 inhibitors are potent CFTR function inducers in PDIOs where residual CFTR function is either present, or created by additional compound exposure. Additionally, upon CFTR modulator treatment we show rescue of CF genotypes that are currently not eligible for this therapy. CONCLUSION: This study exemplifies the feasibility of high-throughput compound screening using PDIOs. We show the potential of repurposing drugs for pwCF carrying non-F508del genotypes that are currently not eligible for therapies. ONE-SENTENCE SUMMARY: We screened 1400 FDA-approved drugs in CF patient-derived intestinal organoids using the previously established functional FIS assay, and show the potential of repurposing PDE4 inhibitors and CFTR modulators for rare CF genotypes.

Maximizing agricultural reuse of recycled nutrients: A spatially explicit assessment of environmental consequences and costs.

Recovering nutrients from organic materials to reduce artificial fertilizer inputs requires the implementation of processing technologies and can involve considerable logistics and transportation costs. Reducing such costs by directly applying organic materials to agricultural land can contribute to pollution due to potential contaminants and unbalanced nutrient ratios. Assessing the cost of increased recycling requires a spatially explicit approach because availability of organic materials, nutrient demand and agro-ecosystem properties vary spatially. A multi-objective model was developed to estimate the trade-offs between costs of nutrient recovery and improvements in nutrient distribution for a case study area in The Netherlands. The evaluated recovery processes included solid-liquid separation followed by reverse osmosis to recover nutrients from pig manure which was compared to a conventional process via hygienisation and export. Results indicate that, even in a nutrient saturated area, replacement potential of artificial nitrogen (N) and phosphorus (P) fertilizers through locally reclaimed nutrients is limited to about 17% N and 55% P. A cost optimum was found when about 48% of the initial pig manure quantities were processed via nutrient recovery and directed to land. Increasing manure processing for nutrient recovery led to a redistribution of nutrients and trace metals (zinc (Zn) and copper (Cu)), resulting in more localized concentration. Zn and Cu were enriched by about 8% and 2%, respectively, when maximizing nutrient recovery. Our generic model offers a methodology to assess the trade-offs between increased recycling and associated spatial effects to facilitate sustainable recycling infrastructures for achieving more circular agriculture.

Bioaccessibility of vitamin A, vitamin C and folic acid from dietary supplements, fortified food and infant formula.

In the Netherlands, vitamin intake occurs mainly via food and for some vitamins also via fortified food. In addition, some people take dietary supplements. Information on the bioavailability of vitamins is important for a good estimation of the actual exposure to vitamins. Furthermore, for a reliable intake estimation, it is important to know the accurateness of the claimed vitamin concentration on the product label. In the current study, the amount of vitamin A, vitamin C, and folic acid in different products and their maximum bioavailability (bioaccessibility) were investigated. In about half of the products, the amount of vitamins significantly deviated from the declared amounts. The vitamin bioaccessibility ranged from <1% to 100%. When assessing the dietary intake exposure of vitamins, it is important to take into account both the possible deviation from the declared level and (the variability of) the bioaccessibility of the vitamin in the products.

Normalization of the auditory startle reflex after symptom reduction in children with anxiety disorders.

BACKGROUND: In an earlier study the Auditory Startle Response (ASR) of anxiety disordered (AD) children proved to be enlarged. This study examines in a controlled design to what extent this increase is responsive to symptom reduction during Cognitive Behavioral Therapy (CBT) METHODS: The activity of 6 muscles following 104 dB tones in 20 patients (M = 12,7 years; SD = 2.5) and 25 matched controls was measured with an electromyogram (EMG). In addition, the sympathetic skin response was investigated. Response to treatment was investigated with the Anxiety Disorder Interview Schedule for Children (ADIS-C) and the Spence Children's Anxiety Scale (SCAS). RESULTS: Treatment responders (n = 12) showed a significant ASR decrease over time, whereas non-responders (n = 8) showed a significant ASR increase or no significant ASR difference. In controls, the ASR was not significantly different at follow up compared to baseline. The sympathetic skin response was stable in controls and treatment responders but significantly increased over time in treatment non-responders. Linear regression suggested that one of the ASR pre-treatment parameters (multiple muscle EMG magnitude) predicts treatment response. CONCLUSIONS: The ASR decreases in AD children when anxiety symptoms diminish. In addition, the ASR may be useful in predicting response to CBT in AD children.

Adaptation of fine roots to annual fertilization and irrigation in a 13-year-old Pinus pinaster stand.

Effects of fertilization and irrigation on fine roots and fungal hyphae were studied in 13-year-old maritime pine (Pinus pinaster Aït. in Soland), 7 years after the initiation of the treatments. The fertilization trials consisted of a phosphorus treatment, a complete fertilizer treatment (N, P, K, Ca and Mg), and an unfertilized treatment (control). Fertilizers were applied annually and were adjusted according to foliar target values. Two irrigation regimes (no irrigation and irrigation of a set amount each day) were applied from May to October. Root samples to depths of 120 cm were collected in summer of 2005, and the biomass of small roots (diameter 2-20 mm) and fine roots (diameter

Recent advances in functional neuroimaging of gait.

In this review, we discuss the contribution of functional neuroimaging to the understanding of the cerebral control of gait in humans, both in healthy subjects and in patients with Parkinson's disease. We illustrate different approaches that have been used to address this issue, ranging from the imaging of actual gait performance to the study of initiation and imagery of gait. We also consider related approaches focused on specific aspects of gait, like those addressed by repetitive foot movements. We provide a critical discussion of advantages and disadvantages of each approach, emphasizing crucial issues to be addressed for a better understanding of the neural control of human gait.

Epidemiology of leprosy on five isolated islands in the Flores Sea, Indonesia

We conducted a population-based survey on 5 small island in South Sulawesi Province (Indonesia) to collect baseline data previous to a chemoprophylactic intervention study aiming at interrrupting the transmission of Mycobacterium leprae. Here we describe the present leprosy epidemiology on these geographically isolated islands. Of the 4774 inhabitants living in the study area 4140 were screened for leprosy (coverage: 87%). We identified 96 leprosy patients (85 new and 11 old patients), representing a new case detection rate (CDR) of 205/10,000 and prevelence rate 195/10,000. CDRs were similar for males and females. Male patients were more often classified as multibacillary (MB) than women. Of the new patients, 33 (39%) were classified as MB, 16 (19%) as paucibacillary (PB) 2-5 lesions and 36 (42%) as PB single lesion. In this area of high leprosy endemicity leprosy patients were extensively clustered, i.e. not equally distributed among the islands and within the islands among the houses

Faecal bacterial profile, nitrogen excretion and mineral absorption in healthy dogs fed supplemental oligofructose.

In a cross-over trial, five healthy dogs were fed a dry food without or with 1% (w/w) oligofructose to assess any oligofructose-induced effects on the faecal bacterial profile, nitrogen excretion and mineral absorption. The diets were given for a period of 3 weeks. Oligofructose feeding significantly raised the number of Bifidobacteria, Streptococci and Clostridia in faeces. The numbers of faecal anaerobic and aerobic bacteria were raised after ingestion of oligofructose. The faecal pH was unchanged. There was no effect of oligofructose feeding on the route of nitrogen excretion which was associated with a lack of effect on faecal ammonium and urinary urea excretion. It is suggested that the absence or presence of an effect of oligofructose on urinary and faecal nitrogen excretion depends on the background composition of the diet, in particular the content of non-digestible, fermentable carbohydrates. In the diets used, the content of non-digestible, fermentable carbohydrates was not measured. Both apparent magnesium and calcium absorption were significantly raised by oligofructose feeding, but phosphorus absorption was unaffected. The data presented may contribute to the qualification of the use of oligofructose in dog foods.

Localization of deposited polycyclic aromatic hydrocarbons in leaves of Plantago.

After deposition to foliage, polycyclic aromatic hydrocarbons (PAHs) may remain on the leaf surface, accumulate in the cuticular wax, or diffuse into the remaining interior of the plant. In a field study, the location of deposited PAHs in the leaves of two Plantago species was determined. To this aim, leaves of Plantago major and Plantago media were divided into three fractions. First, the leaves were washed (wash-off fraction), then cuticular wax was extracted (wax fraction). Finally, the remaining leaf material was extracted (interior fraction). The presence of PAHs could be demonstrated in all three fractions. For both plants, the distribution of PAHs over the three fractions changed with molecular weight (mol wt) of the PAHs. The wash-off fraction increased with increasing molecular weight, likely because high molecular-weight PAHs occur predominantly bound to particles, which can be readily washed off from the leaves. In contrast, the amount of PAHs detected in the interior of the leaves decreased with increasing molecular weight. This can be explained by a slow desorption of the PAHs from the particles and a low diffusion rate of the larger molecules. This study shows that washing reduces the amount of high molecular-weight PAHs on plant surfaces. Therefore, washing of leafy vegetables is important to minimize human dietary intake of PAHs.

Polycyclic aromatic hydrocarbons in soil and plant samples from the vicinity of an oil refinery.

Soil samples, and samples of leaves of Plantago major (great plantain) and grass (mixed species) were collected from the vicinity of an oil refinery in Zelzate, Belgium, and analysed for seven polycyclic aromatic hydrocarbons (PAHs). The samples from the site adjacent to the refinery (site 1) contained very high total PAH-concentrations: namely 300, 8 and 2 microg/g dry wt. for soil, P. major and grass, respectively. Concentrations in samples from more remote sites (up to 4 km from the refinery) were a factor of 10-30 lower than those from site 1, but between them the differences were small. The PAH-profiles of the plant samples, in contrast with those of the soil samples, appeared to shift to higher contributions of gaseous PAHs with increasing distance from the refinery. This can be explained by particle-bound PAHs being deposited closer to the source than gaseous PAHs. It is suggested that particle-bound deposition is relatively more important for deposition to soil than to plants, due to blow-off and wash-off of the compounds from the leaves. The total PAH-concentrations in the leaves of P. major were higher than those measured in the grass samples, probably due to differences in aerodynamic surface roughness, leaf orientation and/or leaf age. However, the concentration ratios of P. major/grass were not constant for the different sites, varying from 1.2 to 8.8. Therefore, it appears that a precise prediction of PAH-concentrations for one plant species from known concentrations of another species is not possible. When errors in predicted concentrations need to be smaller than a factor of approximately 10, the sampling strategy has to be focussed on all species of interest.

A G-->A transition creates a branch point sequence and activation of a cryptic exon, resulting in the hereditary disorder neurofibromatosis 2.

We describe a G-->A transition within intron 5 of the NF2 gene. This mutation creates a consensus splice branch point sequence. To our knowledge this is the first report of a mutation that creates a functional branch point sequence in a human hereditary disorder. The new branch point sequence is located 18 bp upstream of a consensus splice acceptor site. A consensus splice donor site is found 106 bp 3' of the acceptor site. Asa consequence the G-->A transition results in an alternatively spliced mRNA containing an additional exon 5a of 106 bp derived from intron sequences. We cloned the mutant cDNA and show that due to an in-frame stop codon the cDNA codes for a truncated NF2 protein. The mutation was observed in three affected members of an NF2 family. In a tumour of one of the family members both alternatively spliced and wild-type mRNA were found, although the wild-type allele of the gene is absent due to an interstitial deletion on chromosome 22. We also show that immunoprecipitations reveal the presence of full-length wild-type NF2 protein in the tumour lysate. These data support the hypothesis that some degree of normal splicing of the mutant precursor RNA is taking place. It is therefore likely that this residual activity of the mutant allele explains the relatively mild phenotype in the family. These data also indicate that complete inactivation of the gene is not required for tumour formation.

Antiproteinuric effect of ciclosporin A in adriamycin nephropathy in rats.

Ciclosporin A (CsA) can reduce proteinuria in various forms of human and experimental glomerulopathies. This antiproteinuric effect can be the result of a decrease of immunological damage, a decrease in the glomerular filtration rate (GFR), or a change in the permselective properties of the glomerular capillary wall. In this study we investigated the effect of CsA on Adriamycin-induced nephropathy in rats. A single intravenous injection of Adriamycin (5 mg/kg body weight) induced a severe nephrotic syndrome with a massive albuminuria (+/- 400 mg/24 h from 3 weeks onwards) and a hypoalbuminemia (+/- 7 mg/ml after 5 weeks). The IgG/albumin selectivity index was 0.16 +/- 0.05, indicating a preferential loss of albumin. A 5-day treatment with CsA reduced the albumin excretion by almost 50% (from 336 +/- 91 to 178 +/- 58 mg/24 h; p = 0.002) and induced an increase in the serum albumin level (from 7.1 +/- 4.1 to 12.8 +/- 3.2 mg/ml; p = 0.002) in contrast to the vehicle olive oil (OO). CsA also decreased the GFR by 40% (from 0.74 +/- 0.11 to 0.41 +/- 0.11 mg/ml/100 g body weight; p = 0.002). Albuminuria corrected for the GFR (fractional excretion of albumin, FE(alb)) was still significantly lower in CsA-treated than in OO-treated animals (FE(alb) CsA: 1.35 +/- 0.88, FE(alb) OO: 3.17 +/- 2.29%; p = 0.0005). This suggests that other factors are also involved in the reduction of albuminuria. To exclude that CsA has an effect on the tubular reabsorption of albumin, we evaluated the blockade of the tubular reabsorption by lysine and found no difference in albuminuria between the CsA- and OO-treated groups. These experiments suggest that the antiproteinuric effect of CsA is not (only) due to a decrease in the GFR, but also to a decrease of the enhanced permeability of the glomerular capillary wall for albumin.

Influence of a natural-ingredient diet containing Phaseolus vulgaris on the colonization by segmented, filamentous bacteria of the small bowel of mice.

The appearance of segmented, filamentous bacteria (SFBs) in the small bowel of mice is influenced by the composition of the diet, but the dietary components responsible are not known. The addition of ground, whole Phaseolus vulgaris to a natural-ingredient diet at the expense of part of the skim milk powder, ground barley and wheat middlings components, caused an increase of the colonization of the mouse small bowel by SFBs. This effect was not seen when whole Phaseolus was added to a purified diet at the expense of part of the casein, corn oil, coconut fat, corn starch, dextrose and cellulose components. In an attempt to identify the fraction of Phaseolus that might contain SFB-inducing substances, the skin and kernel fraction of the bean were added to the natural-ingredient diet. The skin and kernel fraction were found to be as effective in inducing SFB appearance as was whole Phaseolus.

Bone mineral density after adjuvant chemotherapy for premenopausal breast cancer.

Lumbar bone mineral density (BMD) determination by dual photon absorptiometry was used to study the influence of adjuvant chemotherapy for premenopausal breast cancer on the risk of premature osteoporosis. Six cycles of combination chemotherapy caused ovarian failure in 31 of 44 (71%) women, amenorrhoea mostly already beginning during treatment. In contrast, only seven of 44 (16%) women, who were pair-matched for age and year of breast cancer surgery and had not been treated with chemotherapy, were post-menopausal at the time of measurement. The mean interval after breast surgery was 3.5 years. The significantly decreased BMD in the treated group (1.17 compared to 1.29 g cm-2) could only be explained by the high incidence of menopause in these women, which on average occurred 10 years prematurely. Extrapolation of these findings suggests that adjuvant chemotherapy may precipitate osteoporotic fractures by some 10 years in a considerable proportion of women cured of premenopausal breast cancer.