5-methylthioribose 1-phosphate: a product of partially purified, rat liver 5'-methylthioadenosine phosphorylase activity.

5'-Methylthioadenosine phosphorylase from rat liver has been purified 112-fold. A molecular weight of 90 000 for the enzyme was estimated from gel filtration on Sephadex G-150. The Km for 5'-methylthioadenosine was 4.7 . 10(-7) M, while the Km for phosphate was 2 . 10(-4) M. The products of the reaction were isolated and identified as adenine and 5-methylthioribose 1-phosphate. In addition to 5'-methylthioadenosine the nucleoside analogues 5'-ethylthioadenosine and 5'-n-propylthioadenosine also served as substrates for the enzyme. The 7-deaza analogue 5'-methylthiotubercidin was found to be an inhibitor of the reaction, but was inactive as a substrate.     

Effects of growth hormone treatment on cognitive function and head circumference in children born small for gestational age

Short stature is not the only problem faced by children born small for gestational age (SGA). Being born SGA has also been associated with lowered intelligence, poor academic performance, low social competence and behavioural problems. This paper summarizes the results of a randomized, double-blind, growth hormone (GH) dose-response study (1 or 2 mg/m2/day [ approximately 0.035 or 0.07 mg/kg/day]) on growth, intelligence quotient (IQ) and psychosocial functioning in 79 children born SGA at the start, and after 2 and 8 years of GH therapy, and addresses the associations with head circumference. Mean age at start of therapy was 7.4 years; mean duration of GH treatment was 8.0 years. In 2001, 91% of children born SGA had reached a normal height (> -2.0 standard deviation score [SDS]). Block-design s-score (Performal IQ) and Total IQ score increased (p < 0.001 for both indices) from scores significantly lower than those of Dutch peers at the start of therapy (p < 0.001) to scores that were comparable to those of Dutch peers in 2001. Vocabulary s-score (Verbal IQ) was normal at the start of therapy and remained so over time. Externalizing Problem Behaviour SDS and Total Problem Behaviour SDS improved during GH therapy (p < 0.01-0.05) to scores comparable to those of Dutch peers. Internalizing Problem Behaviour SDS was comparable to that of Dutch peers at the start of therapy and remained so, whereas Self-Perception improved from the start of GH therapy until 2001 (p < 0.001), when it reached normal scores. Head circumference SDS at the start of GH therapy and head growth during GH therapy were positively related to all IQ scores (p < 0.01), whereas neither were related to height SDS at the start of, or to its improvement during, GH therapy. A significant improvement in height and head circumference in children born SGA was seen after only 3 years of GH therapy, in contrast to randomized SGA controls. In conclusion, most children born SGA showed a normalization of height during GH therapy and, in parallel to this, a significant improvement in Performal IQ and Total IQ. In addition, problem behaviour and self-perception improved significantly. Interestingly, Performal, Verbal and Total IQ scores were positively related to head circumference, both at the start of, and during, GH therapy; head circumference increased in GH-treated children born SGA, but not in untreated SGA controls. These results are encouraging but also warrant confirmational studies and further investigations into the effects of GH on the central nervous system.     

Mutations of tubulin glycylation sites reveal cross-talk between the C termini of alpha- and beta-tubulin and affect the ciliary matrix in Tetrahymena

Two types of polymeric post-translational modifications of alpha/beta-tubulin, glycylation and glutamylation, occur widely in cilia and flagella. Their respective cellular functions are poorly understood. Mass spectrometry and immunoblotting showed that two closely related species, the ciliates Tetrahymena and Paramecium, have dramatically different compositions of tubulin post-translational modifications in structurally identical axonemes. Whereas the axonemal tubulin of Paramecium is highly glycylated and has a very low glutamylation content, the axonemal tubulin of Tetrahymena is glycylated and extensively glutamylated. In addition, only the alpha-tubulin of Tetrahymena undergoes detyrosination. Mutations of the known glycylation sites in Tetrahymena tubulin affected the level of each polymeric modification type in both the mutated and nonmutated subunits, revealing cross-talk between alpha- and beta-tubulin. Ultrastructural analyses of glycylation site mutants uncovered defects in the doublet B-subfiber of axonemes and revealed an accumulation of dense material in the ciliary matrix, reminiscent of intraflagellar transport particles seen by others in Chlamydomonas. We propose that polyglycylation and/or polyglutamylation stabilize the B-subfiber of outer doublets and regulate the intraflagellar transport.     

Kombinowanie: agency,informality, and the poetics of self in Highland Poland

This article asks what role local models of agency and subjectivity play in the structure and interpretation of informal economic practices, and how this influences the ways in which people narrate their behaviour to others. Exploring the use of a common Polish idiom for informal activities – kombinowanie – amongst villagers in the Carpathian Highlands, it argues that the term acts as a ‘master‐trope’ for local identity, providing villagers with a narrative model for asserting and broadcasting their social agency to peers. Furthermore, it shows how in this particular ethnographic context, the retelling of instances of informal or illegal activity as kombinowanie can be seen as part of a Highlander ‘poetics of self’, in that each manifestation of the term carries with it a series of implicit sociohistorical and normative meanings that relate to notions of ‘typical’ Górale (Highland) behaviour. By characterizing kombinowanie as a type of tactics, I open up the term for comparative analysis with other local conceptions of agency which value insubordination.     

Characterization of a multiple particle delivery complex and determination of cellular photodamage in skin fibroblast and breast cancer cell lines

Zinc metallized Phthalocyanine (ZnPcS_mix), a potent photosensitizer, is conjugated to gold dendrimer encapsulated nanoparticles (AuDENPs) in order to improve the efficacy of photodynamic therapy (PDT) using MCF‐7 breast cancer cells and WS1 fibroblast cells as a control. Both ZnPcS_mix and AuDENPs are mixed in a nitrogen atmosphere for 48 hours and characterization analysis conducted using ultraviolet‐visible (UV‐vis) spectrometry for spectral properties, transmission electron microscopy (TEM) for morphological features and zeta potential measurement for surface stability and size distribution of the compound obtained or of the multiple particles delivery complex (MPDC). Cell viability, proliferation and membrane damage following PDT are assessed by the trypan blue exclusion test, adenosine triphosphate luminescence and lactate dehydrogenase cytotoxicity assays, respectively. Stable MPDCs are spherical shaped with a diameter lesser than 5 nm, and have a maximum absorption peak at 676 nm. The MPDC‐mediated PDT induces a decrease in cell viability and proliferation, and increased membrane damage or cytotoxicity. The conjugation enhances the therapeutic efficiency of PDT by improving drug delivery and targeting of MCF‐7 cancer cells.      

Photobiomodulation alters matrix protein activity in stressed fibroblast cells in vitro

A balance is maintained between matrix synthesis and degradation, and a prolonged increase in matrix metalloproteinases (MMPs) affects healing. Photobiomodulation (PBM) speeds up healing and alters wound environment. The study aimed to determine changes in protein and gene expression of collagen type 1 (Col‐I), MMP‐3 and ‐9 and TIMP‐1 in fibroblasts irradiated at 660 or 830 nm. Commercially purchased human skin fibroblast cells were modeled into five groups namely, normal, normal wounded, diabetic wounded, hypoxic wounded and diabetic hypoxic wounded. Control cells were sham irradiated. Laser irradiation was conducted at 660 or 830 nm (108/or 94 mW, 9.1 cm², 420/or 483 s) with 5 J/cm². Forty‐eight hours post‐irradiation, protein expression of TIMP‐1, MMP‐3, ?9 and Col‐I was determined by flow cytometry and immunofluorescence, and gene expression by real‐time RT‐PCR. There was an increase in TIMP‐1 and Col‐I, and a decrease in MMP‐3 and ‐9, as well as an alteration in mRNA expression of MMP3, MMP9, TIMP1 and COL1A1 in irradiated cells. Due to the responsiveness of the diabetic hypoxic wounded model, the findings propose this model as appropriate for wound healing studies and suggest that PBM promotes the remodeling phase of wound healing by decreasing matrix degradation and upregulating synthesis.      

Fusions between green fluorescent protein and β-glucuronidase as sensitive and vital bifunctional reporters in plants

By fusing the genes encoding green fluorescent protein (GFP) and -glucuronidase (GUS) we have created a set of bifunctional reporter constructs which are optimized for use in transient and stable expression studies in plants. This approach makes it possible to combine the advantage of GUS, its high sensitivity in histochemical staining, with the advantages of GFP as a vital marker. The fusion proteins were functional in transient expression studies in tobacco using either DNA bombardment or potato virus X as a vector, and in stably transformed Arabidopsis thaliana and Lotus japonicus plants. The results show that high level of expression does not interfere with efficient stable transformation in A. thaliana and L. japonicus. Using confocal laser scanning microscopy we show that the fusion constructs are very suitable for promoter expression studies in all organs of living plants, including root nodules. The use of these reporter constructs in the model legume L. japonicus offers exciting new possibilities for the study of the root nodulation process.     

Acetylcholine-gated current translates wake neuronal firing rate information into a spike timing-based code in Non-REM sleep,stabilizing neural network dynamics during memory consolidation

Sleep is critical for memory consolidation, although the exact mechanisms mediating this process are unknown. Combining reduced network models and analysis of in vivo recordings, we tested the hypothesis that neuromodulatory changes in acetylcholine (ACh) levels during non-rapid eye movement (NREM) sleep mediate stabilization of network-wide firing patterns, with temporal order of neurons’ firing dependent on their mean firing rate during wake. In both reduced models and in vivo recordings from mouse hippocampus, we find that the relative order of firing among neurons during NREM sleep reflects their relative firing rates during prior wake. Our modeling results show that this remapping of wake-associated, firing frequency-based representations is based on NREM-associated changes in neuronal excitability mediated by ACh-gated potassium current. We also show that learning-dependent reordering of sequential firing during NREM sleep, together with spike timing-dependent plasticity (STDP), reconfigures neuronal firing rates across the network. This rescaling of firing rates has been reported in multiple brain circuits across periods of sleep. Our model and experimental data both suggest that this effect is amplified in neural circuits following learning. Together our data suggest that sleep may bias neural networks from firing rate-based towards phase-based information encoding to consolidate memories.