Risk stratification by residual enzyme activity after newborn screening for medium-chain acyl-CoA dehyrogenase deficiency: Data from a cohort study

ABSTRACT: BACKGROUND: Since the introduction of medium-chain acyl coenzyme A dehydrogenase (MCAD) deficiency in population newborn bloodspot screening (NBS) programs, subjects have been identified with variant ACADM (gene encoding MCAD enzyme) genotypes that have never been identified in clinically ascertained patients. It could be hypothesised that residual MCAD enzyme activity can contribute in risk stratification of subjects with variant ACADM genotypes. METHODS: We performed a retrospective cohort study of all patients identified upon population NBS for MCAD deficiency in the Netherlands between 2007-2010. Clinical, molecular, and enzymatic data were integrated. RESULTS: Eighty-four patients from 76 families were identified. Twenty-two percent of the subjects had a variant ACADM genotype. In patients with classical ACADM genotypes, residual MCAD enzyme activity was significantly lower (median 0%, range 0-8%) when compared to subjects with variant ACADM genotypes (range 0-63%; 4 cases with 0%, remainder 20-63%). Patients with (fatal) neonatal presentations before diagnosis displayed residual MCAD enzyme activities <1%. After diagnosis and initiation of treatment, residual MCAD enzyme activities <10% were associated with an increased risk of hypoglycaemia and carnitine supplementation. The prevalence of MCAD deficiency upon screening was 1/8,750 (95% CI 1/7,210-1/11,130). CONCLUSIONS: Determination of residual MCAD enzyme activity improves our understanding of variant ACADM genotypes and may contribute to risk stratification. Subjects with variant ACADM genotypes and residual MCAD enzyme activities <10% should be considered to have the same risks as patients with classical ACADM genotypes. Parental instructions and an emergency regimen will remain principles of the treatment in any type of MCAD deficiency, as the effect of intercurrent illness on residual MCAD enzyme activity remains uncertain. There are, however, arguments in favour of abandoning the general advice to avoid prolonged fasting in subjects with variant ACADM genotypes and 10% residual MCAD enzyme activity.     

Prevention and treatment of tail biting in weaned piglets

The aims of this study were to evaluate four preventive measures and two curative treatments of tail biting. The preventive measures were: chain, rubber hose, straw rack (5 g/pig/day) and the provision of straw on the floor twice daily by hand (2 × 10 g/pig/day). The two curative treatments, which were applied following the onset of tail biting in a pen were: straw twice daily (as in the fourth preventive measure) and the removal of the biter. In total, 960 undocked weaned piglets (10 piglets per pen) were observed during 5 weeks. Tail lesions (none, bite marks and wounds) were recorded daily. The incidence of pens with wounded pig tails was significantly lower when straw was provided twice daily (8% of pens) compared to the chain (58% of pens) and rubber hose (54% of pens) treatment, but did not differ significantly from the straw rack treatment (29% of pens). Tails with bite marks were significantly less common in pens with twice daily straw (16% of pens) compared to chain (88% of pens), rubber hose (79% of pens) and straw rack (75% of pens). No significant difference was found between the curative treatments. Both treatments showed a reduced incidence of red fresh blood on the tails at days 1–9 following curative treatment, compared to day 0. However, neither curative treatment eliminated tail biting entirely. In conclusion, this study indicates that tail biting is best prevented with a small amount of straw, provided twice daily, and to a lesser extent with a straw rack, compared to providing a chain or a rubber hose. Once tail biting has occurred, providing a small amount of straw twice daily and removing the biter appears to be equally effective.     

Regulatory properties and interaction of the C- and N-terminal domains of BetP, an osmoregulated betaine transporter from Corynebacterium glutamicum

The glycine betaine carrier BetP from Corynebacterium glutamicum responds to changes in external osmolality by regulation of its transport activity, and the C-terminal domain was previously identified to be involved in this process. Here we investigate the structural requirements of the C-terminal domain for osmoregulation as well as interacting domains that are relevant for intramolecular signal transduction in response to osmotic stress. For this purpose, we applied a proline scanning approach and amino acid replacements other than proline in selected positions. To analyze the impact of the surrounding membrane, BetP mutants were studied in both C. glutamicum and Escherichia coli, which strongly differ in their phospholipid composition. A region of approximately 25 amino acid residues within the C-terminal domain with a high propensity for alpha-helical structure was found to be essential in terms of its conformational properties for osmodependent regulation. The size of this region was larger in E. coli membranes than in the highly negatively charged C. glutamicum membranes. As a novel aspect of BetP regulation, interaction of the C-terminal domain with one of the cytoplasmic loops as well as with the N-terminal domain was shown to be involved in osmosensing and/or osmoregulation. These results support a functional model of BetP activation that involves the C-terminal domain shifting from interaction with the membrane to interaction with intramolecular domains in response to osmotic stress.     

Comparison of Models Describing Light Dependence of N2 Fixation in Heterocystous Cyanobacteria

The abilities of four models to describe nitrogenase light-response curves were compared, using the heterocystous cyanobacterium Nodularia spumigena and a cyanobacterial bloom from the Baltic Sea as examples. All tested models gave a good fit of the data, and the rectangular hyperbola model is recommended for fitting nitrogenase-light response curves. This model describes an enzymatic process, while the others are empirical. It was possible to convert the process parameters between the four models and compare N₂ fixation with photosynthesis. The physiological meanings of the process parameters are discussed and compared to those of photosynthesis.     

A theoretical approach to predicting the success of genetic manipulation of malaria mosquitoes in malaria control

Background  

Mosquitoes that have been genetically modified to better encapsulate the malaria parasite Plasmodium falciparum are being considered as a possible tool in the control of malaria. Hopes for this have been raised with the identification of genes involved in the encapsulation response and with advances in the tools required to transform mosquitoes. However, we have only very little understanding of the conditions that would allow such genes to spread in natural populations.     

Somaclonal variation rate evolution in plant tissue culture: Contribution to understanding through a statistical approach

Summary In order to better understand somaclonal variant rate evolution in plant tissue culture, a statistical approach has been adopted. According to this approach, the variant percentage could be calculated by: %V=[1−(1−p) ⁿ ]×100, where %V is the percentage of variant, p the probability of variation and n the number of multiplication cycles. A numerical estimation was performed to characterize the variance of this function. It has been demonstrated that a wide scale of variance is associated with ‘%V’, due to the occurrence of variations after a variable number of multiplication cycles in the different lines of culture. Two main conclusions can be drawn from this model: (1) a variant rate increase can be expected as an exponential function of the number of multiplication cycles; (2) after a given number of multiplication cycles, variable off-types percentages can be expected. Due to the complexity of biological systems, this statistical approach could obviously not be applied directly for the calculation and forecasting of variant rates in tissue culture. However, this approach results in a better understanding of two apparently confusing experimental features often reported in tissue culture: the increase of the variant rate as a function of the length of the culture period on the one hand, and, on the other hand, the observations of different variant rates among lines cultured for the same lengths of time under strietly identical culture conditions. This approach also underlined that the comparison of somaclonal variant percentage between batches of plants from different in vitro treatments could be, in some cases, insufficient for ascertaining a difference of variability generated by tissue culture.     

TTP, a C3H zinc finger protein gene, is expressed in mouse ovarian oocytes

The gene TTP, encoding a C3H zinc finger protein of the TIS11 family, is expressed in growing mouse oocytes. The gene is downregulated in Graafian follicles shortly before ovulation. This corresponds to a possible function in regulation of maternal mRNA translation, a function attributed to related C3H class genes in Caenorhabditis elegans, zebrafish, and Xenopus.     

Preparative variant of the purification of a trypsin-like thrombolytic enzyme from Actinomyces 771

A simple large-scale purification scheme for a trypsin-like enzyme from Actinomyces 771 was developed using carboxylic cation exchange resin Soloze K and DEAE-cellulose. The electrophoretically homogeneous enzyme with the specific trypsin activity of 1.4 U/mg was obtained with a yield of 53%.     

Relationship between incurrent flow, calcium ion concentration and membrane potential

A dependence of the inward current across the cell membrane giant neurones at garden snail was investigated under voltage champ. It has been concluded that there are two components of the inward current: a calcium-dependent and I0. The latter current probably was carried by sodium ions. The inward Ca current (ICa) is then given as a function [Ca]2+ by: formula (see text) : KCa is a dissociation constant of the sites in outer part channel independent of membrane voltage. The experimental data are interpreted by two barrier membrane model bases of absolute reaction rate Eyring's theory.