Low compliance with recommendations on folic acid use in relation to pregnancy: is there a need for fortification?

OBJECTIVE: As a means to prevent neural tube defects (NTDs), women planning pregnancy in Denmark are recommended to take a dietary supplement of 400 microg folic acid daily during the periconceptional period. We examined compliance with this recommendation in a national survey. DESIGN: Cohort study on pregnant women in Denmark. SETTING: The Danish National Birth Cohort (DNBC). SUBJECTS: From November 2000 to February 2002, 22,000 pregnant women were recruited for DNBC. Use of dietary supplements was recorded at enrollment. Compliance with the recommendation was related to an information campaign that took place during the second half of 2001, and to lifestyle factors provided in a telephone interview by the end of the first trimester of pregnancy. RESULTS: An increase was seen in the proportion of women complying with the recommendation in the study period and this coincided with the information campaign events. However, even at the end of the period, only 22.3% of the women who had planned their pregnancy fully complied with the recommendation. No increase at all was seen in periconceptional folic acid use among women with unplanned pregnancies. Young age, low education and smoking were identified as factors that determined non-compliance. CONCLUSIONS: Alternative and more effective strategies are needed if the Danish population is to benefit fully from the knowledge that folic acid prevents NTDs. Future strategies should not only target vulnerable groups, such as the less educated and the young, but also women who get pregnant without planning this. The only possible way to reach the last group may be through fortification of foods with folic acid.     

Cloning and analysis of the gene for a major surface antigen of Mycoplasma gallisepticum

Myplasma gallisepticum infects a wide variety of gallineaceous birds including chickens, turkeys, and pheasants. Infection occurs both horizontally and vertically. Thus, control of the spread of M. gallisepticum to noninfected flocks is difficult. Continual monitoring is necessary to identify infected flocks even under the most stringent infectious control practices. Monitoring, however, is usually performed by measuring hemagglutination activity (HA) in serum, an insensitive and variable test. Variability in the HA test arises differences in agglutination antigen, changes in antigenic profiles of the M. gallisepticum strain, and variability in reading the agglutination reaction. Enzyme-linked immunosorbent assays (ELISAs) are the preferred method of testing because of the ease in obtaining sera and the sensitivity and reproducibility of the assays, but the ELISA suffers from a lack of standardization in the test antigen. The ELISA test will be more easily accepted once the test antigen has been standardized. To this end, we have identified, cloned, and characterized the gene for an antigen that has potential as a species-specific antigen for M. gallisepticum The gene codes for a 75-kD protein, P75, that is recognized during natural infections. Recombinant P75 is not recognized in immunoblots by convalescent sera produced in chickens infected with Mycoplasma synoviae, Mycoplasma gallinarum, and Mycoplasma gallinaceum or in turkeys infected with Mycoplasma meleagridis.     

Effect of two-week heat stress during grain filling on stem reserves,senescence, and grain yield of European winter wheat cultivars

To examine the extent to which heat stress during grain filling impacts on the development and yield of winter wheat (Triticum aestivum L.), a 3-year field experiment was conducted on a loess soil with high water holding capacity in the North German Plain. Thirty-two mostly European winter wheat cultivars were exposed to heat stress in a mobile foil tunnel with maximum air temperatures of 45.7, 45.4, and 47.2°C in 2015, 2016, and 2017, respectively. The 14-day post-anthesis heat stress treatment caused an average 57.3% grain yield reduction compared to a close-by non-stressed control. The proportion of green crop area after the heat stress phase varied from 7% to 98% in 2016 and from 37% to 94% in 2017. The green crop area percentage did not significantly correlate with grain yield, indicating that the delayed senescence of stay-green phenotypes offers no yield advantage under terminal heat stress. The water soluble carbohydrate (WSC) concentration of the stems at crop maturity varied between 6 and 92 g/kg dry matter, showing that the genotypes differed in their efficiency at using the stem carbohydrate reserves for grain filling under heat stress. The stem WSC concentration correlated positively with the beginning of anthesis (r = 0.704; p < .001) but negatively with the grain yield (r = −0.431; p < .05). For heat tolerance breeding, the stem reserve strategy, i. e. the rapid and full exhaustion of the temporary carbohydrate storage therefore seems more promising than the stay-green strategy.     

Study of Structure–Activity Relationships of the Marine Alkaloid Fascaplysin and Its Derivatives as Potent Anticancer Agents

Marine alkaloid fascaplysin and its derivatives are known to exhibit promising anticancer properties in vitro and in vivo. However, toxicity of these molecules to non-cancer cells was identified as a main limitation for their clinical use. Here, for the very first time, we synthesized a library of fascaplysin derivatives covering all possible substituent introduction sites, i.e., cycles A, C and E of the 12H-pyrido[1-2-a:3,4-b’]diindole system. Their selectivity towards human prostate cancer versus non-cancer cells, as well as the effects on cellular metabolism, membrane integrity, cell cycle progression, apoptosis induction and their ability to intercalate into DNA were investigated. A pronounced selectivity for cancer cells was observed for the family of di- and trisubstituted halogen derivatives (modification of cycles A and E), while a modification of cycle C resulted in a stronger activity in therapy-resistant PC-3 cells. Among others, 3,10-dibromofascaplysin exhibited the highest selectivity, presumably due to the cytostatic effects executed via the targeting of cellular metabolism. Moreover, an introduction of radical substituents at C-9, C-10 or C-10 plus C-3 resulted in a notable reduction in DNA intercalating activity and improved selectivity. Taken together, our research contributes to understanding the structure–activity relationships of fascaplysin alkaloids and defines further directions of the structural optimization.     

Heavy loads of parasitic freshwater pearl mussel (Margaritifera margaritifera L.) larvae impair foraging,activity and dominance performance in juvenile brown trout (Salmo trutta L.)

The life cycle of the endangered freshwater pearl mussel (Margaritifera margaritifera) includes a parasitic larval phase (glochidia) on the gills of a salmonid host. Glochidia encystment has been shown to affect both swimming ability and prey capture success of brown trout (Salmo trutta), which suggests possible fitness consequences for host fish. To further investigate the relationship between glochidia encystment and behavioural parameters in brown trout, pairs (n = 14) of wild‐caught trout (infested vs. uninfested) were allowed to drift feed in large stream aquaria and foraging success, activity, agonistic behaviour and fish coloration were observed. No differences were found between infested and uninfested fish except for in coloration, where infested fish were significantly darker than uninfested fish. Glochidia load per fish varied from one to several hundred glochidia, however, and high loads had significant effects on foraging, activity and behaviour. Trout with high glochidia loads captured less prey, were less active and showed more subordinate behaviour than did fish with lower loads. Heavy glochidia loads therefore may negatively influence host fitness due to reduced competitive ability. These findings have implications not only for management of mussel populations in the streams, but also for captive breeding programmes which perhaps should avoid high infestation rates. Thus, low levels of infestation on host fish which do not affect trout behaviour but maintains mussel populations may be optimal in these cases.     

Screening Mangrove Endophytic Fungi for Antimalarial Natural Products

We conducted a screening campaign to investigate fungi as a source for new antimalarial compounds. A subset of our fungal collection comprising Chinese mangrove endophytes provided over 5000 lipophilic extracts. We developed an accelerated discovery program based on small-scale cultivation for crude extract screening and a high-throughput malaria assay. Criteria for hits were developed and high priority hits were subjected to scale-up cultivation. Extracts from large scale cultivation were fractionated and these fractions subjected to both in vitro malaria and cytotoxicity screening. Criteria for advancing fractions to purification were developed, including the introduction of a selectivity index and by dereplication of known metabolites. From the Chinese mangrove endophytes, four new compounds (14–16, 18) were isolated including a new dimeric tetrahydroxanthone, dicerandrol D (14), which was found to display the most favorable bioactivity profile.     

Sampling downed coarse woody debris in fire-prone eucalypt woodlands

Downed coarse woody debris (DCWD) plays an important role in ecosystem processes and should be considered in land management decisions. Unfortunately accurate quantification of its abundance is difficult, due to its patchy distribution. This is especially problematic in woodland ecosystems where DCWD is relatively scarce, unevenly distributed and smaller in size than in other forest types. This study compared the efficacy of the line intersect and strip plot methods to sample properties of DCWD at woodland sites with differing fire histories. Although measures of abundance using the two methods did not differ statistically, the line intersect method had 20% less variability in the data, was quicker to perform and made it easier to locate individual pieces of DCWD than the strip plot method. The results of this study indicate that transects of 100 m or less are insufficient for estimating DCWD volume in woodlands. An acceptable level of precision was reached at approximately 450 m at very recently burnt sites and 700 m at long unburnt sites when only transect length was considered. However when taking both transect length and number of replicates into account, an acceptable level of precision was reached at 500 m when sampling a minimum of 20 sites. It is therefore recommended that pilot studies should be conducted to determine appropriate sampling intensities in previously unsampled areas as DCWD volume estimates are sensitive to the rate at which variance changes with increasing transect length. This is particularly important when there is variation in the disturbance history of the sites. However, if it is not possible to conduct pilot studies, the requirement would be to sample line intersect transects of at least 500 m, at a minimum of 20 sites in woodland systems.     

Identification and stereochemical characterization of lignans in flaxseed and pumpkin seeds

Phytoestrogens of the lignan type are widely distributed in plant-derived food items and are believed to protect against hormone-dependent cancer. The richest known dietary source of lignans is flaxseed. Flaxseed has been reported to contain glycosides of secoisolariciresinol as the major lignan, together with small amounts of matairesinol, isolariciresinol, and pinoresinol. Secoisolariciresinol, but none of the other lignans, has so far been identified in pumpkin seeds. In the present study, two different methods for the hydrolysis of lignan glycosides are compared. Artifact formation and loss of lignans under acidic conditions were observed. Lariciresinol was identified by GC-MS analysis in two different types of flaxseed (Linum usitatissimum L. and Linum flavum L.) and in pumpkin seeds (Cucurbita pepo L.) for the first time. Likewise, the novel lignan demethoxy-secoisolariciresinol was tentatively identified in the flaxseed samples. Stereochemical analysis by chiral HPLC of several lignans isolated from flaxseed showed that secoisolariciresinol, matairesinol, and lariciresinol consisted predominantly of one enantiomer.     

Isolation and characterisation of Mycobacterium avium and Rhodococcus equi from granulomatous lesions of swine lymph nodes in Slovenia

Granulomatous lesions in bovine and especially swine lymph nodes are still frequently observed during routine veterinary meat inspections even though Mycobacterium bovis infections are no longer detected in domestic animals in Slovenia. Different lymph nodes of pigs (n = 260) were investigated using classical bacteriological and molecular methods. Mycobacterium avium alone was isolated in 47.3% of pigs and in mixed infection with Rhodococcus equi in 3.9% of pigs. R. equi alone was isolated in 27.3% and in mixed infection with mycobacteria other than M. avium in 1.5% of pigs. A total of 133 M. avium isolates were typed using the IS1245, IS901 and FR300 PCR. Almost two thirds (60.9%) of isolates belonged to M. avium hominissuis (IS901-, IS1245+ genotype), 33.8% of isolates belonged to M. avium avium (IS901+, IS1245+ genotype) and 5.3% of isolates remained non-typed. Fifty out of 85 R. equi isolates were tested for the virulence-associated antigens (VapA and VapB). Nearly two thirds (60.0%) were positive for VapB while all the other isolates were VapA- and VapB-negative.