Developmental regulation of pectic epitopes during potato tuberisation

We show, by immunogold labelling, that potato (Solanum tuberosum L. cv Karnico) pectic epitopes are developmentally regulated within regions of the stolon, in addition to showing tissue-specific differences in abundance and localisation. The (1-->4)-beta-D-galactan and (1-->5)-alpha-arabinan epitopes demarcate two distinct zones within stolons; galactans are enriched in primary walls of elongating cells proximal to the stolon hook, whilst arabinans predominate in younger cells distal to the hook. Low-methoxyl homogalacturonan epitopes are concentrated in the middle lamella and show a proximo-distal gradient in stolons similar to that of galactans, whilst high-methoxyl homogalacturonan is uniformly abundant. Calcium pectate is restricted to the middle lamella at cell corners and pit fields. Calcium-binding sites are uniformly present in stolon cell walls, but their total density is reduced and they become localised to a few cell corners in mature tubers, as determined by image-electron energy loss spectroscopy. During the transition from elongation growth to isodiametric expansion during tuberisation of the stolon hook, there were no detectable changes in pectic epitope abundance or localisation. As tubers matured, all epitopes increased in abundance in parenchymal cell walls, except for calcium pectate. We conclude that potentially significant changes in pectic composition occur as young cells distal to the stolon hook move into the zone of cell elongation proximal to the hook.     

Effects of vitamin B12 supplementation on neurodevelopment and growth in Nepalese Infants: A randomized controlled trial

BackgroundVitamin B₁₂ deficiency is common and affects cell division and differentiation, erythropoiesis, and the central nervous system. Several observational studies have demonstrated associations between biomarkers of vitamin B₁₂ status with growth, neurodevelopment, and anemia. The objective of this study was to measure the effects of daily supplementation of vitamin B₁₂ for 1 year on neurodevelopment, growth, and hemoglobin concentration in infants at risk of deficiency.Methods and findingsThis is a community-based, individually randomized, double-blind placebo-controlled trial conducted in low- to middle-income neighborhoods in Bhaktapur, Nepal. We enrolled 600 marginally stunted, 6- to 11-month-old infants between April 2015 and February 2017. Children were randomized in a 1:1 ratio to 2 μg of vitamin B₁₂, corresponding to approximately 2 to 3 recommended daily allowances (RDAs) or a placebo daily for 12 months. Both groups were also given 15 other vitamins and minerals at around 1 RDA. The primary outcomes were neurodevelopment measured by the Bayley Scales of Infant and Toddler Development 3rd ed. (Bayley-III), attained growth, and hemoglobin concentration. Secondary outcomes included the metabolic response measured by plasma total homocysteine (tHcy) and methylmalonic acid (MMA). A total of 16 children (2.7%) in the vitamin B₁₂ group and 10 children (1.7%) in the placebo group were lost to follow-up. Of note, 94% of the scheduled daily doses of vitamin B₁₂ or placebo were reported to have been consumed (in part or completely). In this study, we observed that there were no effects of the intervention on the Bayley-III scores, growth, or hemoglobin concentration. Children in both groups grew on an average 12.5 cm (SD: 1.8), and the mean difference was 0.20 cm (95% confidence interval (CI): −0.23 to 0.63, P = 0.354). Furthermore, at the end of the study, the mean difference in hemoglobin concentration was 0.02 g/dL (95% CI: −1.33 to 1.37, P = 0.978), and the difference in the cognitive scaled scores was 0.16 (95% CI: −0.54 to 0.87, P = 0.648). The tHcy and MMA concentrations were 23% (95% CI: 17 to 30, P < 0.001) and 30% (95% CI: 15 to 46, P < 0.001) higher in the placebo group than in the vitamin B₁₂ group, respectively. We observed 43 adverse events in 36 children, and these events were not associated with the intervention. In addition, 20 in the vitamin B₁₂ group and 16 in the placebo group were hospitalized during the supplementation period. Important limitations of the study are that the strict inclusion criteria could limit the external validity and that the period of vitamin B₁₂ supplementation might not have covered a critical window for infant growth or brain development.ConclusionsIn this study, we observed that vitamin B₁₂ supplementation in young children at risk of vitamin B₁₂ deficiency resulted in an improved metabolic response but did not affect neurodevelopment, growth, or hemoglobin concentration. Our results do not support widespread vitamin B₁₂ supplementation in marginalized infants from low-income countries.Trial registrationClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT02272842","term_id":"NCT02272842"}}NCT02272842Universal Trial Number: U1111-1161-5187 (September 8, 2014)Trial Protocol: Original trial protocol: PMID: 28431557 (reference [18]; study protocols and plan of analysis included as Supporting information).

Tor A. Strand and colleagues measure the effects of daily supplementation of vitamin B12 for one year on neurodevelopment, growth, and hemoglobin concentration in infants at risk of deficiency.     

Updated Risk/Benefit Analysis of Fish Consumption Effects on Neurodevelopment: Implications for Setting Advisories

Epidemiological studies indicate that fish consumption during pregnancy leads to improved neurodevelopment. This suggests that the beneficial nutrients in fish may offset the adverse effects of mercury in the case of the average fish meal. However, our previous risk/benefit model predicted a net neurodevelopmental risk for the majority of species analyzed. In this article the previous model is calibrated against fish benefit data and then compared to other fish risk/benefit models, including recent models from the World Health Organization and USFDA. Our calibrated model estimated greater benefit for low mercury species but greater risk for high mercury species than the other models. With respect to a commonly eaten high mercury fish, swordfish, the calibrated model yielded risks that are supportive of current fish advisories but, in contrast, the other models predicted net neurodevelopmental benefits. The calibrated model was used in a proposed 3 step framework for setting fish consumption advisories: (1) Set initial consumption level based upon mercury RfD; (2) Adjust consumption upward if risk/benefit model indicates a net benefit; (3) Cap fish consumption based upon saturation of O-3 benefit. The implications of this approach for 7 varieties of fish are used to illustrate the framework.     

A rapid method to screen for cell-wall mutants using discriminant analysis of Fourier transform infrared spectra

We have developed a rapid method to screen large numbers of mutant plants for a broad range of cell wall phenotypes using Fourier transform infrared (FTIR) microspectroscopy of leaves. We established and validated a model that can discriminate between the leaves of wild-type and a previously defined set of cell-wall mutants of Arabidopsis . Exploratory principal component analysis indicated that mutants deficient in different cell-wall sugars can be distinguished from each other. Discrimination of cell-wall mutants from wild-type was independent of variability in starch content or additional unrelated mutations that might be present in a heavily mutagenised population. We then developed an analysis of FTIR spectra of leaves obtained from over 1000 mutagenised flax plants, and selected 59 plants whose spectral variation from wild-type was significantly out of the range of a wild-type population, determined by Mahalanobis distance. Cell wall sugars from the leaves of selected putative mutants were assayed by gas chromatography-mass spectrometry and 42 showed significant differences in neutral sugar composition. The FTIR spectra indicated that six of the remaining 17 plants have altered ester or protein content. We conclude that linear discriminant analysis of FTIR spectra is a robust method to identify a broad range of structural and architectural alterations in cell walls, appearing as a consequence of developmental regulation, environmental adaptation or genetic modification.     

An analysis of Ca2+ release by DGEA: mobilization of two functionally distinct internal stores in Saos-2 cells

Osteoblasts can be activated by their collagen matrix and inparticular the DGEA peptide motif. We have reported that DGEA is ableto activate Ca²⁺ signalingpathways in the human osteoblast-like cell line, Saos-2, by a tyrosinekinase-dependent pathway (T. J. McCann, W. T. Mason, M. C. Meikle, andF. McDonald. Matrix Biol. 16:271-280, 1997). In the present study, we show that this activityis due to coupling of the signal to intracellularCa²⁺ stores, since the DGEA actionis not blocked by La³⁺ but is lostwhen Ca²⁺ stores are depleted with2 µM and blocked by 10 µM ryanodine. The activated stores alsodiffer functionally from those activated by thrombin, as blockade withU-73122 obstructs only thrombin-activated Ca²⁺ release. We have shown thatthe DGEA activity was not due to its high-charge density, since the twoacidic residues can be substituted with their uncharged homologues(asparagine and glutamine) without significant loss of activity. Thiswas in turn measured by an adhesion assay that also demonstrated thislevel of specificity. Furthermore, by constructing DGEA bound to FITC,we have shown that DGEA binding was dependent on divalent cations. Wehave also demonstrated that an intact actin cytoskeleton is notrequired for Ca²⁺ activation byinhibiting actin polymerization with the addition of cytochalasin B. These data strengthen the argument that collagen has a significant rolein regulating osteoblast function via this peptide motif.

     

Stimulation of prolactin secretion by morphine: role of the central serotonergic system.

Unanesthetized male rats with indwellinh right atrial cannulae were injected with morphine (MOR) i.v. which produced a dose-related increase in plasma prolactin levels (PRL). This effect was blocked partially by naloxone (NAL) at a dose of 0.06 mg/kg and totally by 0.6 mg/kg NAL. Interruption of central serotonergic neurotransmission by receptor blockade, with metergoline (MET) or cyproheptadine (Cypro), inhibition of tryptophan hydroxylase by para-chlorophenylalanine or destruction of serotonin neurons by 5, 7-dihydroxytryptamine antagonized the morphine (3 mg/kg) induced elevation in PRL release. Depression of dopaminergic activity with α-methyl-para-tyrosine elevated the basal PRL levels, but it did not prevent a further increase of prolactin levels by morphine (3 mg/kg). These data are compatible with the hypothesis that morphine stimulates PRL release by activation of the central serotonergic system.     

Further studies on prostaglandin E2 (PGE2)-induced gonadotropin release: Comparison with the effect of 15-methyl PGE2, PGAs, PGBs and prostaglandin endoperoxide analogs

It is known that PGE₂ is a potent stimulus of LH release. To determine if the effect of PGE₂ could be enhanced and/or prolonged by retarding its metabolic degradation, a derivative, 15-methyl PGE₂ (15-E₂) which is more slowly degraded than the natural compound was injected intravenously (i.v.) at various dose levels or into the third ventricle (3rd V) of ether-anesthetized, ovariectomized, estrogen (OVX, Eb)-treated rats and its effect on gonadotropin release was compared with that of PGE₂. Both PGs injected i.v. were equally effective in increasing plasma LH and maintaining the elevated levels, although 15-E₂ induced a larger and more sustained increase in plasma FSH than PGE₂. By contrast, 3rd V PGE₂ was clearly more effective than 3rd V 15-E₂ in releasing LH and to a lesser extent, FSH. The effect of 15-E₂ on LH was similar to that produced by 3rd V PGE₁ injected at a similar dose. However, its effect on FSH was greater than that of PGE₁.To evaluate the effect(s) of prostaglandins of the A and B series on gonadotropin release, PGA₁, PGA₂, PGB₁ or PGB₂ were injected intraventricularly in OVX, Eb-treated rats. PGBs were injected into conscious, free-moving rats. PGA₂ or PGB₂ increased plasma LH concnetrations although much less effectively than PGE₂. Third V PGA₁ or PGB₁ were ineffective. The 3rd V injection of two cyclic esters (U-44069 and U-46619), stable analogs of the PG endoperoxide PGG₂ and PGH₂, induced a small, transient increase in LH levels and did not alter plasma FSH in conscious, free-moving animals. PGE₂ injected intraventricularly at a similar dose was demonstrated to be much more potent than the analogs in stimulating LH and FSH release. The results indicate that: 1) 15-E₂, in spite of its described long-lasting activity, does not appear to be more potent than the natural compound in releasing LH, although when injected i.v., it appeared to induce a more sustained increase in plasma FSH; 2) although PGA₂ and PGB₂ can also act centrally to stimulate LH release, their low potency suggests that this is a pharmacological effect; and 3) the two analogs of PG endoperoxides tested proved to be poor stimuli for gonadotropin release. The significance of these findings is discussed.