Molecular and morphological characterization of monosomic additions in Beta vulgaris,carrying extra chromosomes of B. procumbens or B. patellaris

DNA fingerprinting with three repetitive DNA sequences (OPX2, PB6-4 and Sat-121) was carried out on a set of 10 monosomic additions of Beta procumbens and 75 anonymous B. patellaris-derived monosomic additions in B. vulgaris, for characterization of the alien chromosomes at the DNA level. The probes are Procumbentes-specific and distributed over all chromosomes. Morphological characteristics were also used for the classification of B. patellaris monosomic addition families and for comparison with the morphology of the addition families of B. procumbens.DNA fingerprinting revealed unique patterns for almost all individual addition chromosomes of B. procumbens. However, it was concluded that chromosomes 1 and 6 of B. procumbens may be identical with the only difference that the chromosome referred to as 6 carries a susceptible allele for beet cyst nematode (BCN) resistance. In contrast, it was concluded that the two addition types with chromosome 2 are carrying different chromosomes of B. procumbens, so that one of them was renumbered to become the new chromosome 6.DNA fingerprinting of 75 anonymous B. patellaris-derived monosomic additions facilitated the identification and characterization of the alien chromosomes and the grouping of these additions into nine different groups. Several of these groups could be divided in two sub-groups on the basis of small differences in banding patterns. The results of the DNA fingerprinting led to the conclusion that B. patellaris most likely is an allotetraploid. It was also deduced that the BCN gene(s) in this species are homozygous and located on chromosome 1, while the pair of homoeologous chromosomes does not carry such BCN gene(s). Because of the allotetraploid nature of B. patellaris, preferential association occurs between the two homologous chromosomes containing the allele(s) for BCN resistance. Each group of B. patellaris addition families united by DNA fingerprinting had comparable morphological characteristics. Some of these morphological traits appeared to be chromosome-specific and were very useful for primary classification of the addition families. However, the present study showed that these morphological traits are not adequate for the identification of all alien chromosomes without the aid of additional markers. Because of similarities observed between molecular characteristics or the effects on plant morphology of several chromosomes of B. procumbens and B. patellaris it was concluded that B. procumbens could have been involved in the evolutionary history of B. patellaris.     

Targeted disruption of the insulin receptor gene in the mouse results in neonatal lethality.

Targeted disruption of the insulin receptor gene (Insr) in the mouse was achieved using the homologous recombination approach. Insr+/- mice were normal as shown by glucose tolerance tests. Normal Insr-/- pups were born at expected rates, indicating that Insr can be dispensable for intrauterine development, growth and metabolism. However, they rapidly developed diabetic ketoacidosis accompanied by a marked post-natal growth retardation (up to 30-40% of littermate size), skeletal muscle hypotrophy and fatty infiltration of the liver and they died within 7 days after birth. Total absence of the insulin receptor (IR), demonstrated in the homozygous mutant mice, also resulted in other metabolic disorders: plasma triglyceride level could increase 6-fold and hepatic glycogen content could be five times less as compared with normal littermates. The very pronounced hyperglycemia in Insr-/- mice could result in an increased plasma insulin level of up to approximately 300 microU/ml, as compared with approximately 25 microU/ml for normal littermates. However, this plasma level was still unexpectedly low when compared with human infants with leprechaunism, who lack IR but who could have extremely high insulinemia (up to > 4000 microU/ml). The pathogenesis resulting from a null mutation in Insr is discussed.     

Cadabicine,an alkaloid from Cadaba farinosa

A new spermidine alkaloid, cadabicine, has been isolated from Cadaba farinosa. Its structure was determined by X-ray crystallographic and spectral methods.     

Toward the development of smart and low cost point-of-care biosensors based on screen printed electrodes

Screen printing technology provides a cheap and easy means to fabricate disposable electrochemical devices in bulk quantities which are used for rapid, low-cost, on-site, real-time and recurrent industrial, pharmaceutical or environmental analyses. Recent developments in micro-fabrication and nano-characterization made it possible to screen print reproducible feature on materials including plastics, ceramics and metals. The processed features forms screen-printed disposable biochip (SPDB) upon the application of suitable bio-chemical recognition receptors following appropriate methods. Adequacy of biological and non-biological materials is the key to successful biochip development. We can further improve recognition ability of SPDBs by adopting new screen printed electrode (SPE) configurations. This review covers screen-printing theory with special emphasis on the technical impacts of SPE architectures, surface treatments, operational stability and signal sensitivity. The application of SPE in different areas has also been summarized. The article aims to highlight the state-of-the-art of SPDB at the laboratory scale to enable us in envisaging the deployment of emerging SPDB technology on the commercial scale.     

Morphological Description and Molecular Phylogeny of Two Species of Levicoleps (Ciliophora,Prostomatida), L. taehwae nov. spec. and L. biwae jejuensis nov. subspec., collected in Korea

Two colepid ciliates, Levicoleps taehwae nov. spec. and L. biwae jejuensis nov. subspec., were collected from the brackish water of the Taehwa River and a small freshwater pond in Jeju Island, South Korea, respectively. Their living morphology, infraciliature, and small subunit (SSU) rRNA gene sequences were determined using standard methods. Barrel‐shaped L. taehwae nov. spec. is a small ciliate with an average size of 45 × 25 μm in vivo, about 15 ciliary rows each composed of 12 monokinetids and two perioral dikinetids, and two 20 μm‐long caudal cilia. The sequence length and GC content of the SSU rRNA gene are 1,669 bp, 44.5%. This novel species is similar in body size to Coleps hirtus, and has six armor tiers and hirtus‐type tier plates, and the same number of ciliary rows as C. hirtus; however, it can be distinguished from the latter by the absence of armor spines and its sequence similarity of SSU rRNA gene is about 92.8% which indicates that it is a distinct form. Levicoleps biwae jejuensis nov. subspec., is a medium colepid ciliate which has a barrel‐shaped body, about 22 somatic kineties and 16 transverse ciliary rows, three mini adoral organelles, and four 15 μm‐long caudal cilia. The sequence length and GC content of the SSU rRNA gene are 1,666 bp and 44.4%.     

Characterizing monoclonal antibody formulations in arginine glutamate solutions using 1H NMR spectroscopy

Assessing how excipients affect the self-association of monoclonal antibodies (mAbs) requires informative and direct in situ measurements for highly concentrated solutions, without sample dilution or perturbation. This study explores the application of solution nuclear magnetic resonance (NMR) spectroscopy for characterization of typical mAb behavior in formulations containing arginine glutamate. The data show that the analysis of signal intensities in 1D ¹H NMR spectra, when compensated for changes in buffer viscosity, is invaluable for identifying conditions where protein-protein interactions are minimized. NMR-derived molecular translational diffusion rates for concentrated solutions are less useful than transverse relaxation rates as parameters defining optimal formulation. Furthermore, NMR reports on the solution viscosity and mAb aggregation during accelerated stability study assessment, generating data consistent with that acquired by size-exclusion chromatography. The methodology developed here offers NMR spectroscopy as a new tool providing complementary information useful to formulation development of mAbs and other large therapeutic proteins.