Halogenated derivatives of boldine with high selectivity for alpha1A-adrenoceptors in rat cerebral cortex

The selectivity of 3-nitrosoboldine and different halogenated derivatives of boldine (3-bromoboldine, 3,8-dibromoboldine and 3-chloroboldine) for alpha1-adrenoceptor subtypes was studied by examining [3H]-prazosin competition binding in rat cerebral cortex. In the competition experiments [3H]-prazosin binding was inhibited completely by all the compounds tested. The inhibition curves displayed shallow slopes which could be subdivided into high and low affinity components. The relative order of affinity and selectivity for alpha1A-adrenoceptors was 3-bromoboldine = 3,8-dibromoboldine = 3-chloroboldine > boldine > 3-nitrosoboldine. The competition curves for 3-bromoboldine remained shallow and biphasic following chloroethylclonidine treatment. Whereas the relative contribution of the high affinity sites increased, the 3-bromoboldine affinities at its high and low affinity sites remained similar to those obtained in untreated membranes. 3-Bromoboldine, 3,8-dibromoboldine, 3-chloroboldine and 3-nitrosoboldine did not significantly displace [3H]-(+)-cis-diltiazem binding to rat cerebral cortex membranes. This activity was lower than that shown by boldine. Compared to boldine, halogen (bromine or chlorine) substitution at position 3 increases the alpha1A-adrenoceptor subtype selectivity and decreases the affinity for the benzothiazepine binding site at the calcium channel. Further halogen substitution at position 8 did not significantly improve this activity with respect to 3-bromoboldine. In contrast, the NO substitution at position 3 of boldine (3-nitrosoboldine) gives a loss of affinity and selectivity for alpha1-adrenoceptor subtypes.     

Astragaloside IV Protects against Isoproterenol-Induced Cardiac Hypertrophy by Regulating NF-κB/PGC-1α Signaling Mediated Energy Biosynthesis

We previously reported that Astragaloside IV (ASIV), a major active constituent of Astragalus membranaceus (Fisch) Bge protects against cardiac hypertrophy in rats induced by isoproterenol (Iso), however the mechanism underlying the protection remains unknown. Dysfunction of cardiac energy biosynthesis contributes to the hypertrophy and Nuclear Factor κB (NF-κB)/Peroxisome Proliferator-Activated Receptor-γ Coactivator 1α (PGC-1α) signaling gets involved in the dysfunction. The present study was designed to investigate the mechanism by which ASIV improves the cardiac hypertrophy with focuses on the NF-κB/PGC-1α signaling mediated energy biosynthesis. Sprague-Dawley (SD) rats or Neonatal Rat Ventricular Myocytes (NRVMs) were treated with Iso alone or in combination with ASIV. The results showed that combination with ASIV significantly attenuated the pathological changes, reduced the ratios of heart weight/body weight and Left ventricular weight/body weight, improved the cardiac hemodynamics, down-regulated mRNA expression of Atrial Natriuretic Peptide (ANP) and Brain Natriuretic Peptide (BNP), increased the ratio of ATP/AMP, and decreased the content of Free Fat Acid (FFA) in heart tissue of rats compared with Iso alone. In addition, pretreatment with ASIV significantly decreased the surface area and protein content, down-regulated mRNA expression of ANP and BNP, increased the ratio of ATP/AMP, and decreased the content of FFA in NRVMs compared with Iso alone. Furthermore, ASIV increased the protein expression of ATP5D, subunit of ATP synthase and PGC-1α, inhibited translocation of p65, subunit of NF-κB into nuclear fraction in both rats and NRVMs compared with Iso alone. Parthenolide (Par), the specific inhibitor of p65, exerted similar effects as ASIV in NRVMs. Knockdown of p65 with siRNA decreased the surface areas and increased PGC-1α expression of NRVMs compared with Iso alone. The results suggested that ASIV protects against Iso-induced cardiac hypertrophy through regulating NF-κB/PGC-1α signaling mediated energy biosynthesis.     

The Prognostic Role of Ezrin Immunoexpression in Osteosarcoma: A Meta-Analysis of Published Data

Background

The significance of ezrin immunoexpression and prognosis for osteosarcoma is still controversial. The aim was to provide a meta-analysis for ezrin immunoexpression and prognostic features of osteosarcoma patients.

Methods

A detailed search was made in MEDLINE, EMBASE and the Web of Knowledge for relevant original articles published in English; methodological quality of the included studies was also assessed. Two reviewers extracted data independently. Studies were pooled and summary hazard ratios (HRs) and odds ratio (ORs) with corresponding confidence intervals (CIs) were calculated.

Results

Final analysis of 318 patients from 5 eligible studies was performed. Combined HR of ezrin immunohistochemical staining suggested that positive immunoexpression had an unfavorable impact on osteosarcoma patients'' overall survival (n = 223 in 4 studies; HR = 4.79; 95% CI: 1.50–15.30; P = 0.008) but not on event-free survival (n = 202 in 3 studies; HR = 1.59; 95% CI: 0.61–4.15; P = 0. 0.342). Combined OR of ezrin immunohistochemical staining indicated that positive immunoexpression was associated with recurrence (n = 134 in 2 studies; OR = 3.79; 95% CI: 1.49–9.64; P = 0.005) but not with serum ALP level (n = 160 in 2 studies; OR = 2.16; 95% CI: 0.09–52.50; P = 0.637) and histological response to neoadjuvant chemotherapy(n = 260 in 4 studies; OR = 0.87; 95% CI: 0.37–2.03; P = 0.740).

Conclusions

The results of this meta-analysis suggest that ezrin positive immunoexpression confers a higher risk of recurrence and a worse survival in osteosarcoma patients. Large prospective studies are needed to provide solid data to investigate the precise prognostic significance of ezrin.     

Relationships between Growth,Growth Response to Nutrient Supply,and Ion Content Using a Recombinant Inbred Line Population in Arabidopsis

Growth is an integrative trait that responds to environmental factors and is crucial for plant fitness. A major environmental factor influencing plant growth is nutrient supply. In order to explore this relationship further, we quantified growth-related traits, ion content, and other biochemical traits (protein, hexose, and chlorophyll contents) of a recombinant inbred line population of Arabidopsis (Arabidopsis thaliana) grown on different levels of potassium and phosphate. Performing an all subsets multiple regression analyses revealed a link between growth-related traits and mineral nutrient content. Based on our results, up to 85% of growth variation can be explained by variation in ion content, highlighting the importance of ionomics for a broader understanding of plant growth. In addition, quantitative trait loci (QTLs) were detected for growth-related traits, ion content, further biochemical traits, and their responses to reduced supplies of potassium or phosphate. Colocalization of these QTLs is explored, and candidate genes are discussed. A QTL for rosette weight response to reduced potassium supply was identified on the bottom of chromosome 5, and its effects were validated using selected near isogenic lines. These lines retained over 20% more rosette weight in reduced potassium supply, accompanied by an increase in potassium content in their leaves.Plants in natural environments face abiotic constraints limiting growth and ultimately affecting their fitness. In response to such constraints, flowering time (Korves et al., 2007) and seed dormancy (Donohue et al., 2005) as well as vegetative growth (Barto and Cipollini, 2005; Milla et al., 2009) are the main traits controlling fitness (for review, see Alonso-Blanco et al., 2009). These traits are under the control of complex networks integrating genetic (G) and environmental (E) factors as well as their interaction (G × E). Due to the implications for food and renewable energy sources, dissecting the genetic architecture that underlies plant growth is becoming a priority for plant science (Rengel and Damon, 2008; Carroll and Somerville, 2009; Gilbert, 2009).Plant growth is highly dependent on mineral nutrient uptake (Clarkson, 1980; Sinclair, 1992). Minerals can be distinguished into two categories based on the amount required by plants: micronutrients, which are found in relatively small amounts in the plant (such as copper and iron), and macronutrients, which constitute between 1,000 and 15,000 μg g⁻¹ plant dry weight (such as potassium and phosphate; Marschner, 1995, Buchanan et al., 2002). Phosphate is an important structural and signaling molecule with an essential role in photosynthesis, energy conservation, and carbon metabolism. Its deficiency leads to a reduction of growth and an increase of pathogen susceptibility (Marschner, 1995; Williamson et al., 2001; Abel et al., 2002; López-Bucio et al., 2005; Poirier and Bucher, 2008; Vijayraghavan and Soole, 2010). Potassium is not incorporated into any organic substances but acts as the major osmoticum of the cell, controlling cell expansion, plasma membrane potential and transport, pH value, and many other catalytic processes (Maathuis and Sanders, 1996; Armengaud et al., 2004; Christian et al., 2006; Di Cera, 2006). Potassium deficiency leads to reduced plant growth, a loss of turgor, increased susceptibility to cold stress and pathogens, and the development of chlorosis and necrosis (Marschner, 1995; Véry and Sentenac, 2003; Ashley et al., 2006; Amtmann et al., 2008). To cope with changes in nutrient availability, plants have evolved different mechanisms of adaptation, such as changes in ion transporter expression and activity (Ashley et al., 2006; Jung et al., 2009), morphological changes, such as an increase in root growth to explore more soil volume (Marschner, 1995; Shirvani et al., 2001; Jiang et al., 2007; Jordan-Meille and Pellerin, 2008), or acidification of the surrounding soil in order to mobilize more mineral nutrients (for review, see Ryan et al., 2001). Although these adaptations are well known, the mechanisms involved in sensing and signaling low mineral nutrient status are less well understood, despite significant progress in this area being made (Doerner, 2008; Jung et al., 2009; Luan et al., 2009; Wang and Wu, 2010).One approach to identify genes that are involved in plant responses to environmental factors is to perform a quantitative trait locus (QTL) analysis on a mapping population grown in contrasting environments, allowing the identification of QTL-environment (QTL × E) interactions. Some QTLs for growth-related traits in response to environmental changes were cloned already. For example, the differential response of root growth of some Arabidopsis (Arabidopsis thaliana) accessions to phosphate starvation led to the identification of allelic differences responsible for this phenotype (Reymond et al., 2006; Svistoonoff et al., 2007). Other studies have identified QTLs for shoot dry matter under changing nitrogen supply (Rauh et al., 2002; Loudet et al., 2003). In parallel to natural variation for growth, natural variation for ion content has also been reported. In Arabidopsis, considerable variation in the content of mineral nutrients exists both in seeds (Vreugdenhil et al., 2004; Waters and Grusak, 2008) and in leaves (Harada and Leigh, 2006; Rus et al., 2006; Baxter et al., 2008a; Morrissey et al., 2009). Furthermore, changes in mineral nutrient homeostasis have also been reported to be associated with characteristic multivariate changes in the leaf ionome, the mineral nutrient and trace element composition of an organism or an organ (Baxter et al., 2008b). Due to higher throughput and lower costs, such “omics” analyses examining alterations of large numbers of certain molecules at once have recently become available for mapping purposes. Some QTL studies have linked the variations of these omics data to variation of growth or other physiological traits. For instance, Meyer et al. (2007) and Schauer et al. (2008) linked plant growth or morphological traits to a synergistic network of metabolomic compounds in Arabidopsis and tomato (Solanum lycopersicum), respectively. In addition, Sulpice et al. (2009) associated differences in growth with starch content using a set of Arabidopsis accessions. Compiling the importance of ions in the process of cell division (Lai et al., 2007; Sano et al., 2007) or cell expansion (Philippar et al., 1999; Elumalai et al., 2002), ionomics appears to be a major unexplored field for understanding growth.In this study, we focus on variation in plant growth, the root and leaf ionomes, and their response to varying supplies of potassium and phosphate. Studying variations for these traits among recombinant inbred lines (RILs) in Arabidopsis enabled us to detect QTL and QTL × E interactions for all of these traits. To understand the observed variation in plant growth, predictors that explained a high percentage of variation of growth-related traits have been selected especially among the root and leaf ionomes. The colocalization between growth-related trait QTLs and QTLs for their predictors allowed us to point out genetic regions of possible causality. In addition, the effect of a growth-response QTL on reduced potassium supply was validated with selected near isogenic lines (NILs) that maintained a higher rosette weight when grown in reduced potassium supply. This growth advantage went along with significant changes in ion contents that further emphasize the impact of the ionome in plant growth variations.     

Differential effects of propofol and ketamine on critical brain dynamics

Whether the brain operates at a critical “tipping” point is a long standing scientific question, with evidence from both cellular and systems-scale studies suggesting that the brain does sit in, or near, a critical regime. Neuroimaging studies of humans in altered states of consciousness have prompted the suggestion that maintenance of critical dynamics is necessary for the emergence of consciousness and complex cognition, and that reduced or disorganized consciousness may be associated with deviations from criticality. Unfortunately, many of the cellular-level studies reporting signs of criticality were performed in non-conscious systems (in vitro neuronal cultures) or unconscious animals (e.g. anaesthetized rats). Here we attempted to address this knowledge gap by exploring critical brain dynamics in invasive ECoG recordings from multiple sessions with a single macaque as the animal transitioned from consciousness to unconsciousness under different anaesthetics (ketamine and propofol). We use a previously-validated test of criticality: avalanche dynamics to assess the differences in brain dynamics between normal consciousness and both drug-states. Propofol and ketamine were selected due to their differential effects on consciousness (ketamine, but not propofol, is known to induce an unusual state known as “dissociative anaesthesia”). Our analyses indicate that propofol dramatically restricted the size and duration of avalanches, while ketamine allowed for more awake-like dynamics to persist. In addition, propofol, but not ketamine, triggered a large reduction in the complexity of brain dynamics. All states, however, showed some signs of persistent criticality when testing for exponent relations and universal shape-collapse. Further, maintenance of critical brain dynamics may be important for regulation and control of conscious awareness.     

Magnetic Resonance Imaging Assessment of Effective Ablated Volume following High Intensity Focused Ultrasound

Under magnetic resonance (MR) guidance, high intensity focused ultrasound (HIFU) is capable of precise and accurate delivery of thermal dose to tissues. Given the excellent soft tissue imaging capabilities of MRI, but the lack of data on the correlation of MRI findings to histology following HIFU, we sought to examine tumor response to HIFU ablation to determine whether there was a correlation between histological findings and common MR imaging protocols in the assessment of the extent of thermal damage. Female FVB mice (n = 34), bearing bilateral neu deletion tumors, were unilaterally insonated under MR guidance, with the contralateral tumor as a control. Between one and five spots (focal size 0.5 × 0.5 × 2.5 mm³) were insonated per tumor with each spot receiving approximately 74.2 J of acoustic energy over a period of 7 seconds. Animals were then imaged on a 7T MR scanner with several protocols. T1 weighted images (with and without gadolinium contrast) were collected in addition to a series of T2 weighted and diffusion weighted images (for later reconstruction into T2 and apparent diffusion coefficient maps), immediately following ablation and at 6, 24, and 48 hours post treatment. Animals were sacrificed at each time point and both insonated/treated and contralateral tumors removed and stained for NADH-diaphorase, caspase 3, or with hematoxylin and eosin (H&E). We found the area of non-enhancement on contrast enhanced T1 weighted imaging immediately post ablation correlated with the region of tissue receiving a thermal dose CEM43 ≥ 240 min. Moreover, while both tumor T2 and apparent diffusion coefficient values changed from pre-ablation values, contrast enhanced T1 weighted images appeared to be more senstive to changes in tissue viability following HIFU ablation.     

Influence of <Emphasis Type="Italic">Lactobacillus fermentum</Emphasis> I5007 on the intestinal and systemic immune responses of healthy and <Emphasis Type="Italic">E. coli</Emphasis> challenged piglets

The effect of feeding Lactobacillus fermentum I5007 on the immune system of weaned pigs with or without E. coli challenge was determined. Twenty-four weaned barrows (6.07 ± 0.63 kg BW) were randomly assigned to one of four treatments (N = 6) in a factorial design experiment. The first two treatments consisted of healthy piglets with half of the pigs receiving no treatment while the other half was orally administered with L. fermentum I5007 (10⁸ CFU/ml) at a daily dose of 20 ml. Pigs in the second two treatments were challenged on the first day with 20 ml of E. coli K88ac (10⁸ CFU/ml). Half of these pigs were not treated while the remaining pigs were treated with 20 ml of L. fermentum I5007 (10⁸ CFU/ml). Peripheral blood lymphocytes subsets were determined using flow cytometry. The intestinal mucosal immunity of the pigs was monitored by real time polymerase chain reaction. The cytokine content of the pig’s serum was also analyzed. Oral administration of L. fermentum I5007 increased blood CD4⁺ lymphocyte subset percentage as well as tumor necrosis factor-α and interferon-γ expression in the ileum. Pigs challenged with E. coli had elevated jejunal tumor necrosis factor-α while interferon-γ expression was increased throughout the small intestine. There was no difference in the concentration of the cytokines interleukin-2, interleukin-6, tumor necrosis factor-α and interferon-γ in the serum. CD8⁺ and CD4⁺/CD8⁺ in peripheral blood were not affected by treatment. In conclusion, L. fermentum I5007 can enhance T cell differentiation and induce ileum cytokine expression suggesting that this probiotic strain could modulate immune function in piglets.     

Oxidative stress and life histories: unresolved issues and current needs

Life‐history theory concerns the trade‐offs that mold the patterns of investment by animals between reproduction, growth, and survival. It is widely recognized that physiology plays a role in the mediation of life‐history trade‐offs, but the details remain obscure. As life‐history theory concerns aspects of investment in the soma that influence survival, understanding the physiological basis of life histories is related, but not identical, to understanding the process of aging. One idea from the field of aging that has gained considerable traction in the area of life histories is that life‐history trade‐offs may be mediated by free radical production and oxidative stress. We outline here developments in this field and summarize a number of important unresolved issues that may guide future research efforts. The issues are as follows. First, different tissues and macromolecular targets of oxidative stress respond differently during reproduction. The functional significance of these changes, however, remains uncertain. Consequently there is a need for studies that link oxidative stress measurements to functional outcomes, such as survival. Second, measurements of oxidative stress are often highly invasive or terminal. Terminal studies of oxidative stress in wild animals, where detailed life‐history information is available, cannot generally be performed without compromising the aims of the studies that generated the life‐history data. There is a need therefore for novel non‐invasive measurements of multi‐tissue oxidative stress. Third, laboratory studies provide unrivaled opportunities for experimental manipulation but may fail to expose the physiology underpinning life‐history effects, because of the benign laboratory environment. Fourth, the idea that oxidative stress might underlie life‐history trade‐offs does not make specific enough predictions that are amenable to testing. Moreover, there is a paucity of good alternative theoretical models on which contrasting predictions might be based. Fifth, there is an enormous diversity of life‐history variation to test the idea that oxidative stress may be a key mediator. So far we have only scratched the surface. Broadening the scope may reveal new strategies linked to the processes of oxidative damage and repair. Finally, understanding the trade‐offs in life histories and understanding the process of aging are related but not identical questions. Scientists inhabiting these two spheres of activity seldom collide, yet they have much to learn from each other.     

A Comparison between Multiple Regression Models and CUN-BAE Equation to Predict Body Fat in Adults

Background

Because the accurate measure of body fat (BF) is difficult, several prediction equations have been proposed. The aim of this study was to compare different multiple regression models to predict BF, including the recently reported CUN-BAE equation.

Methods

Multi regression models using body mass index (BMI) and body adiposity index (BAI) as predictors of BF will be compared. These models will be also compared with the CUN-BAE equation. For all the analysis a sample including all the participants and another one including only the overweight and obese subjects will be considered. The BF reference measure was made using Bioelectrical Impedance Analysis.

Results

The simplest models including only BMI or BAI as independent variables showed that BAI is a better predictor of BF. However, adding the variable sex to both models made BMI a better predictor than the BAI. For both the whole group of participants and the group of overweight and obese participants, using simple models (BMI, age and sex as variables) allowed obtaining similar correlations with BF as when the more complex CUN-BAE was used (ρ = 0:87 vs. ρ = 0:86 for the whole sample and ρ = 0:88 vs. ρ = 0:89 for overweight and obese subjects, being the second value the one for CUN-BAE).

Conclusions

There are simpler models than CUN-BAE equation that fits BF as well as CUN-BAE does. Therefore, it could be considered that CUN-BAE overfits. Using a simple linear regression model, the BAI, as the only variable, predicts BF better than BMI. However, when the sex variable is introduced, BMI becomes the indicator of choice to predict BF.     

Microhabitat distribution and behaviour of Branchiobdellidan Holtodrilus truncatus found on the freshwater shrimp Neocaridina spp. from the Sugo River, Japan

A study was performed on the microhabitat distribution and some aspects of behaviour of the ectosymbiotic branchiobdellidan Holtodrilus truncatus (Annelida, Clitellata) found on the freshwater shrimp that inhabit the Sugo River, Hyogo Prefecture, western Japan. Observations on shrimp that were collected from the Sugo River (2003 to 2011) confirmed that the host shrimp is Neocaridina spp. (Atyidae). The attachment location on the host shrimp was predominately between the 1^st pleopod and the 5th pereopod (55.3%). The reproductive method of H. truncatus is hemaphroditism. The cocoon was found only inside the carapace of the host shrimp. The cocoon was transparent and contained a maximum of 14 juvenile worms (developing embryos). When hatching approached, H. truncatus’s worms became elongated and slender, and only one worm hatched out at a time. When Holtodrilus truncatus was removed from its host and was maintained in river water without any food, it survived for a maximum of 46 days. In a host exchange experiment, where we provided several other freshwater shrimp species, Palaemonidae fed on H. truncatus. Moreover, Palaemon paucidens and Macrobrachium nipponense from Lake Biwa also preyed upon H. truncatus. The possible symbiotic relationship between H. truncatus and Neocaridina spp. (family Atyidae) is further discussed.