Synthesis of steroid-ferrocene conjugates of steroidal 17-carboxamides via a palladium-catalyzed aminocarbonylation--copper-catalyzed azide-alkyne cycloaddition reaction sequence

Steroids with the 17-iodo-16-ene functionality were converted to ferrocene labeled steroidal 17-carboxamides via a two step reaction sequence. The first step involved the palladium-catalyzed aminocarbonylation of the alkenyl iodides with prop-2-yn-1-amine as the nucleophile in the presence of the Pd(OAc)₂/PPh₃ catalyst system. In the second step, the product N-(prop-2-ynyl)-carboxamides underwent a facile azide–alkyne cycloaddition with ferrocenyl azides in the presence of CuSO₄/sodium ascorbate to produce the steroid–ferrocene conjugates. The new compounds were obtained in good yield and were characterized by ¹H and ¹³C NMR, IR, MS and elemental analysis.     

Influence of galactoglucomannan oligosaccharides on root culture of <Emphasis Type="Italic">Karwinskia humboldtiana</Emphasis>

Galactoglucomannan oligosaccharides (GGMOs) activity in K. humboldtiana root culture has been determined. GGMOs inhibited adventitious root growth and lateral root induction in contrast to IAA, IBA, and NAA stimulating effect in these processes. Similarly, the combination of GGMOs with natural auxins (IAA, IBA) evoked an inhibition of adventitious root growth and lateral root induction that depended on the oligosaccharides concentration and the type of auxin. The growth stimulating effect of the synthetic auxin, NAA, in adventitious roots was negatively affected by GGMOs, but they were without influence on lateral root induction. The presence of oligosaccharides triggered lateral root position on adventitious roots and the anatomy of adventitious roots (diameter, proportion of primary cortex to the central cylinder, number and size of primary cortical cells, intercellular spaces, and the number of starch grains in cells of primary cortex) in dependence on their coactions with auxin.     

A survey of mixed Begomovirus infection in solanaceae and fabaceae at different altitudes in East Java,Indonesia

A multiplex primer set was developed to detect four Begomoviruses in East Java, Indonesia, i.e. Tomato leaf curl New Delhi virus (ToLCNDV), Tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), Pepper yellow leaf curl Indonesia virus (PepYLCIV) and Mungbean yellow mosaic India virus (MYMIV). Survey at different altitudes found that begomoviruses infecting pepper, tomato and long bean were more variable, while in eggplant and string bean were more uniform. As a single virus, TYLCKaV infected eggplant, and sometimes tomato and pepper; PepYLCIV infected pepper, tomato and long bean; ToLCNDV only infected long bean and tomato at low frequency; and MYMIV infected beans. Mixed infection occurred more frequently in the low altitude areas. Subsequent examination indicated that Cucumber mosaic virus (CMV) and potyviruses were also responsible for diseased fabaceous. Our data suggest a relationship between altitudes and virus species occurrence. However, which viral species infects a crop is mainly influenced by the crop rather than by altitude.     

Procedures to genotype problematic non-invasive otter (Lutra lutra) samples

Non-invasive genetics is a powerful tool in wildlife research and monitoring, especially when dealing with elusive and rare species such as the Eurasian otter (Lutra lutra). Nevertheless, otter DNA obtained from scats and anal secretions appears to be exposed to very quick degradation processes, and the success rate in DNA amplification is lower than in other carnivores. We collected 191 samples from April to September 2011 along the river Sangro basin (Italy) which was recently re-colonized by the Eurasian otter. Using two sets of microsatellite loci (six Lut and seven OT loci), we investigated the influence of sample type and age, collection time, storage time, temperature and humidity on genotyping success and amplification success. We also tested the efficacy of different DNA extraction kits and storage buffer mediums. Finally, we compared amplification success rate, allelic dropout and false allele rates for each locus. We obtained a mean amplification success rate of 79.0 % and a genotyping success rate of 35.1 %. Fresh pure jellies yielded the highest amplification success and genotyping rate. Six microsatellite loci should be theoretically sufficient to distinguish the individual unrelated otters (PID?=?0.001), while 13 loci were needed to distinguish sibling otters (PID_sibs?=?0.002) in our population. We identified 11 otters, and molecular sexing ascertained the presence of five males, four females and two uncertain individuals. Generalized linear models highlighted a significant influence of sample type and age, temperature and humidity both on genotyping and amplification success.     

Water stress and abscisic acid treatments induce the CAM pathway in the epiphytic fern Vittaria lineata (L.) Smith

Among various epiphytic ferns found in the Brazilian Atlantic Forest, we studied Vittaria lineata (L.) Smith (Polypodiopsida, Pteridaceae). Anatomical characterization of the leaf was carried out by light microscopy, fluorescence microscopy, and scanning electron microscopy. V. lineata possesses succulent leaves with two longitudinal furrows on the abaxial surface. We observed abundant stomata inside the furrows, glandular trichomes, paraphises, and sporangia. We examined malate concentrations in leaves, relative water content (RWC), photosynthetic pigments, and chlorophyll (Chl) a fluorescence in control, water-deficient, and abscisic acid (ABA)-treated plants. Plants subjected to drought stress (DS) and treated by exogenous ABA showed significant increase in the malate concentration, demonstrating nocturnal acidification. These findings suggest that V. lineata could change its mode of carbon fixation from C₃ to the CAM pathway in response to drought. No significant changes in RWC were observed among treatments. Moreover, although plants subjected to stress treatments showed a significant decline in the contents of Chl a and b, the concentrations of carotenoids were stable. Photosynthetic parameters obtained from rapid light curves showed a significant decrease after DS and ABA treatments.     

Nursing Home Admission during the First Year after Hospitalization – The Contribution of Cognitive Impairment

Objective

The importance of cognitive and physical functioning for nursing home admission among medically hospitalized older patients is rarely studied in a one-year follow-up perspective. This study aims to explore the association between patient characteristics and nursing home admission within one year after hospitalization in persons 65 years or more.

Design

A one-year longitudinal study.

Methods

We included 463 (234 women) persons aged 65 years or more from internal medical wards in a rural area of Norway. Cognitive function was assessed using the Mini Mental State Examination; physical and instrumental functional status was assessed using the physical self-maintenance scale and instrumental activities of daily living scale of Lawton and Brody. Comorbidity was measured with the Charlson index. Admission to nursing home within one year (yes versus no) was analyzed using logistic regression analysis and Cox proportional hazard regression analysis.

Results

The mean age of the sample was 80.5 (SD 7.4) years, mean Mini Mental State Examination score was 24.1 (SD 3.8) (maximum score is 30). In adjusted analysis participants with cognitive impairment (a Mini Mental State Examination score <25) or impaired physical functioning at baseline had higher risk of admission to nursing home within one year (OR 3.0, 95%CI 1.5–6.2 and OR 3.5, 95%CI 1.8–9.6, respectively). The time before admission was also associated with cognitive impairment and impaired physical functioning in the adjusted analysis (HR 2.6 95%CI 1.4–4.8 and HR 3.7, 95%CI 1.5–8.9, respectively).

Conclusion

Impaired cognitive and physical functioning increased the risk for nursing home admission within one year after hospitalization. However, putative regressors, such as education and social network were not included in the analysis.     

An easy method for cutting and fluorescent staining of thin roots

Background and Aims

Cutting plant material is essential for observing internal structures and may be difficult for various reasons. Most fixation agents such as aldehydes, as well as embedding resins, do not allow subsequent use of fluorescent staining and make material too soft to make good-quality hand-sections. Moreover, cutting thin roots can be very difficult and time consuming. A new, fast and effective method to provide good-quality sections and fluorescent staining of fresh or fixed root samples, including those of very thin roots (such as Arabidopsis or Noccaea), is described here.

Methods

To overcome the above-mentioned difficulties the following procedure is proposed: fixation in methanol (when fresh material cannot be used) followed by en bloc staining with toluidine blue, embedding in 6 % agarose, preparation of free-hand sections of embedded material, staining with fluorescent dye, and observation in a microscope under UV light.

Key Results

Despite eventual slight deformation of primary cell walls (depending on the species and root developmental stage), this method allows effective observation of different structures such as ontogenetic changes of cells along the root axis, e.g. development of xylem elements, deposition of Casparian bands and suberin lamellae in endodermis or exodermis or peri-endodermal thickenings in Noccaea roots.

Conclusions

This method provides good-quality sections and allows relatively rapid detection of cell-wall modifications. Also important is the possibility of using this method for free-hand cutting of extremely thin roots such as those of Arabidopsis.     

Outer dynein arm light chain 1 is essential for controlling the ciliary response to cyclic AMP in Paramecium tetraurelia

The individual role of the outer dynein arm light chains in the molecular mechanisms of ciliary movements in response to second messengers, such as Ca(2+) and cyclic nucleotides, is unclear. We examined the role of the gene termed the outer dynein arm light chain 1 (LC1) gene of Paramecium tetraurelia (ODAL1), a homologue of the outer dynein arm LC1 gene of Chlamydomonas reinhardtii, in ciliary movements by RNA interference (RNAi) using a feeding method. The ODAL1-silenced (ODAL1-RNAi) cells swam slowly, and their swimming velocity did not increase in response to membrane-hyperpolarizing stimuli. Ciliary movements on the cortical sheets of ODAL1-RNAi cells revealed that the ciliary beat frequency was significantly lower than that of control cells in the presence of ≥ 1 mM Mg(2+)-ATP. In addition, the ciliary orientation of ODAL1-RNAi cells did not change in response to cyclic AMP (cAMP). A 29-kDa protein phosphorylated in a cAMP-dependent manner in the control cells disappeared in the axoneme of ODAL1-RNAi cells. These results indicate that ODAL1 is essential for controlling the ciliary response by cAMP-dependent phosphorylation.