Isolation and characterization of human urinary colony-stimulating factor

CSF-1 was isolated from a large volume of human normal urine (10,000 l), using the following 5 stages of purification: concentration by dialysis, silica gel adsorption, hydrophobic chromatography on phenyl-Sepharose CL-6B, fast protein liquid chromatography (FPLC) and finally preparative electrophoresis on polyacrylamide gels. We have isolated 8 mg of purified CSF-1 which migrated as a single band under non-reducing conditions in SDS-PAGE (staining with Coomassie Blue and the sensitive silver techniques). But in the presence of dithiothreitol, the SDS-PAGE pattern revealed a minor second band with a molecular mass of 50,000 Da. CSF-1 was purified 100,000-fold and has a specific activity of 2.16 X 10(7) units/mg protein. Its apparent molecular mass is 57,000 Da with an isoelectric point, pI = 5.8-6.0. The amino-acid composition is reported and compared with that of murine CSF-1. The carbohydrate content (sialic acid, sulphate groups, N-acetylglucosamine, N-acetylgalactosamine) was also determined, and it shows that CSF is a glycoprotein.     

The effect of VDR gene polymorphisms and vitamin D level on blood pressure,risk of preeclampsia,gestational age,and body mass index

We investigated the influence of vitamin D receptor (VDR) polymorphisms and vitamin D level on the blood pressure and the risk of preeclampsia. In a case-control study, 200 pregnant women, including 100 individuals with preeclampsia along with 100 healthy pregnant women, were studied for VDR FokI, TaqI, and BmsI polymorphisms and serum 25 (OH)-D level using polymerase chain reaction-restriction fragment length polymorphism method and commercial kit, respectively. The mean level of 25 (OH)-D in preeclamptic patients was significantly lower (16.6 ± 4.2 ng/mL, P < 0.001) compared with controls (19.6 ± 3.8 ng/mL). Among all women, a significantly higher systolic blood pressure and before-pregnancy body mass index and also lower gestational age were observed in the presence of 25 (OH)-D level < 20 ng/mL compared with the 20 to 30 ng/mL. A significantly higher frequency of VDR FokI C allele in preeclamptic patients (83%) than controls (74%) was associated with a 1.72-fold increased risk of preeclampsia. In all the studied individuals, the systolic and diastolic blood pressures were significantly higher in the presence of the FokI CC genotype compared with the TC and TT+TC genotypes. Neither VDR Taq1 nor VDR BmsI was associated with the risk of preeclampsia. The haplotype FokI C, TaqI C and BmsI A (CCA) compared with haplotype CTG increased the risk of preeclampsia by 1.4-fold (P = 0.33). Our study suggests an association between VDR FokI polymorphism and an insufficient serum level of 25 (OH)-D with the risk of preeclampsia and also the influence of insufficient 25 (OH)-D level and VDR FokI polymorphism on maternal factors, including blood pressure.     

Effects of essential fatty acids on the reproduction of a generalist herbivore

We studied direct and indirect effects of essential fatty acids(EFA) on Daphnia magna reproduction. Daphnia females that receivedEFA enriched algae over their entire lifespan produced largerbroods than females fed control algae. However, EFA-enrichedfemales were also 91% heavier than control females, which potentiallyexplain the larger investment in reproduction. Thus, with sucha large difference in female mass, it is difficult to differentiatethe direct effect of the EFA addition from the indirect (maternalsize) effects on reproduction. To assess the direct effectson reproduction, we performed two experiments in which we enrichedfemale diets with a range of fatty acids. To minimize maternalsize differences, we applied the EFA enrichments only to maturedaphniids and studied the effects on reproduction during andafter two time intervals (15–16 and 7 days). Limitingthe enrichment phase until after maturity decreased the maternalsize differences over the enrichment phase among the fatty acidtreatments from an average of 91% (for life time enrichment)to 29% after 15–16 days maternal enrichment interval,to 18% after the 7 day interval. Minimizing size differencesbetween differently enriched females decreased the differencesin brood size and offspring size. Neonates from control femaleswere more severely affected by starvation than offspring fromfemales that received saturated and polyunsaturated fatty acidenrichments. Under low food concentrations, only neonates thathad access to polyunsaturated fatty acids had positive growthrates, thus showing that although dietary fatty acids can beused for energy purposes, specific fatty acids are requiredto build new biomass. One consequence of our findings is thatoffspring size does not serve as a good estimate of offspringquality when feeding on different resource qualities.     

Zooplankton eat what they need: copepod selective feeding and potential consequences for marine systems

Herbivores are generally faced with a plethora of resources which differ in quality. Therefore, they should be able to select foods which most closely match their metabolic needs. Here, we tested the hypothesis that copepods of the species Acartia tonsa select prey cells based on quality differences within prey species. We assessed age‐specific variation in feeding behaviour and evaluated the potential consequences of such variation for nutrient cycles. Nauplii (young) stages characterized by a low nitrogen to phosphorus (N:P) ratio in their body tissue selected for phosphorus‐rich food, while older copepodite stages with higher body N:P selected for nitrogen‐rich food. Further, the analysis of a 35‐year data set in the southern North Sea revealed a positive correlation between the abundance of nauplii and the ratio of dissolved inorganic N:P, thus suggesting that P‐availability for primary producers declines with the population densities of nauplii. Our findings demonstrate that a combination of stage‐specific selective feeding and body stoichiometry has the potential to affect cycling of limiting nutrients when consumer populations change in composition.     

A truncated FatB resulting from a single nucleotide insertion is responsible for reducing saturated fatty acids in maize seed oil

Key message

We identified a G-nucleotide insertion in a maize FatB responsible for reducing saturated fatty acids through QTL mapping and map-based cloning and developed an allele-specific DNA marker for molecular breeding.

Abstract

Vegetable oils with reduced saturated fatty acids have signficant health benefits. SRS72NE, a Dow AgroSciences proprietory maize inbred line, was found to contain signficantly reduced levels of palmitic acid and total saturated fatty acids in seed oil when compared to other common inbreds. Using F₂ and F₃ populations derived from a cross between SRS72NE and a normal inbred SLN74, we have demonstrated that the reduced saturated fatty acid phenotype in SRS72NE is controlled by a single QTL on chromosome 9 that explains 79.1 % of palmitic acid and 79.6 % total saturated fatty acid variations. The QTL was mapped to an interval of 105 kb that contains one single gene, a type B fatty acyl-ACP thioesterase (ZmFatB; GRMZM5G829544). ZmFatB alleles from SRS72NE and common inbreds were cloned and sequenced. SRS72NE fatb allele contains a single nucleotide (G) insertion in the 6th exon, which creates a premature stop codon 22 base pairs down stream. As a result, ZmFatB protein from SRS72NE is predicted to contain eight altered and 90 deleted amino acids at its C-terminus. Because the affected region is part of the conserved acyl-ACP thioesterase catalytic domain, the truncated ZmFatB in SRS72NE is likely non-functional. We also show that fatb RNA level in SRS72NE is reduced by 4.4-fold when compared to the normal allele SNL74. A high throughput DNA assay capable of differentiating the normal and reduced saturate fatty acid alleles has been developed and can be used for accelerated molecular breeding.     

Predator-induced life-history changes and the coexistence of five taxa in a Daphnia species complex

Interspecific hybridization is common in water fleas of the Daphnia galeata species complex (e.g. D. galeata , D. cucullata , D. hyalina and their interspecific hybrids). We studied the effect of fish on the life histories of five taxa of this species complex originating from the Plußsee, northern Germany. Using four clones per taxon, we found that fish kairomones reduce size at birth and size at maturity significantly. For size at maturity larger taxa showed a significantly stronger reaction to fish kairomones than the smaller taxa. With respect to the intrinsic rate of increase, r , we compared two predation regimes (positive size selective and not selective). We found that under the fish predation regime most clones had a higher r when cultured with fish kairomones, leading to a higher r , and a stronger reaction for the smaller taxa. We conclude that fish predation might be an important factor influencing the co-occurrence of Daphnia parental taxa with their hybrids.