Short communication: Circulating and milk adiponectin change differently during energy deficiency at different stages of lactation in dairy cows

Adiponectin, one of the most abundant adipokines in circulation, is known for its role in regulation of body metabolism. The aim of this study was to evaluate the effects of a negative energy balance (NEB) at 2 stages of lactation (lactational NEB at the onset of lactation and an induced NEB by feed restriction near 100 d of lactation) on circulating adiponectin concentrations. We also investigated the effect of feed restriction on adiponectin concentrations in milk and the relationships of blood and milk adiponectin with selected plasma or milk variables and with measures of body condition. Plasma adiponectin was measured in 50 multiparous Holstein dairy cows throughout 3 experimental periods [i.e., period 1=3 wk antepartum up to 12 wk postpartum, period 2=3 wk of feed restriction starting at around 100 d in milk with a control (n=25) and feed-restricted group (50% of energy requirements; n=25), and period 3=subsequent realimentation period for 8 wk]. Milk adiponectin was investigated among 21 multiparous cows at wk 2 and wk 12 of period 1 and wk 2 of period 2. Adiponectin concentrations in plasma and skim milk were measured using an in-house ELISA specific for bovine adiponectin. Major changes in circulating adiponectin concentrations were observed during the periparturient period, whereas energy deficiency during established lactation at around 100 d in milk and subsequent refeeding did not affect plasma adiponectin. Together with lower adiponectin concentrations in milk (µg/mL), the reduction in milk yield led to decreased adiponectin secretion via milk (mg/d) at the second week of feed restriction. Irrespective of time and treatment, milk adiponectin represented about 0.002% of total milk protein. Mean adiponectin concentrations in milk (0.61 ± 0.03 µg/mL) were about 92% lower than the mean plasma adiponectin concentrations (32.1 ± 1.0 µg/mL). The proportion of the steady-state plasma adiponectin pool secreted daily via milk was 2.7%. In view of the similar extent of NEB in both periods of energy deficiency, decreasing adiponectin concentrations seems important for accomplishing the adaptation to the rapidly increasing metabolic rates in early lactation, whereas the lipolytic reaction toward feed restriction-induced NEB during established lactation seems to occur largely independent of changes in circulating adiponectin.

Temporal analysis of PI-3kinase and MAPK signal transduction during skeletal muscle overload

Skeletal muscles can adapt to increased mechanical forces (or loading) by increasing the size and strength of the muscle. Knowledge of the molecular mechanisms by which muscle responds to increased loading may lead to the discovery of novel treatment strategies for muscle wasting and frailty. The objective of this research was to examine the temporal associations between the activation of specific signaling pathway intermediates and their potential upstream regulator(s) in response to increased muscle loading. Previous work has demonstrated that focal adhesion kinase (FAK) activity is increased in overloaded hypertrophying skeletal muscle. Thus FAK is a candidate for transducing the loading stimulus in skeletal muscle, potentially by activating phosphatidylinositol 3-kinase (PI3K) and members of the mitogen-activated protein kinase (MAPK) family. However, it was unknown if muscle overload would result in activation of PI3K or the MAPKs. Thus, this work seeks to characterized the temporal response of (1) MAPK phosphorylation (including Erk 2, p38 MAPK and JNK), (2) PI3K activity, and (3) FAK tyrosine phosphorylation in response to 24 hours of compensatory overload in the rat soleus and plantaris muscles. In both muscles, overload resulted in transient Increases in the phosphorylation state of Erk2 and JNK, which peaked within the first hour of overload and returned to baseline thereafter. In contrast, p38 MAPK phosphorylation remained elevated throughout the entire 24-hour overload period. Moreover, overload increased PI3K activity only, in the plantaris and only at 12 hours. Moreover, 24 hours of overload induced a significant increase in total protein content in the plantaris but not the soleus. Thus an increase in total muscle protein content within the 24-hour loading period was observed only in muscle exhibiting increased PI3K activity. Surprisingly, FAK tyrosine phosphorylation was not increased during the overload period in either muscle, indicating that PI3K activation and increased MAPK phosphorylation were independent of increased FAK tyrosine phosphorylation. In summary, increased PI3K activity and sustained elevation of p38 MAPK phosphorylation were associated with muscle overload, identifying these pathways as potential mediators of the early hypertrophic response to skeletal muscle overload. This suggests that stimuli or mechanisms that activate these pathways may reduce/minimize muscle wasting and frailty. ^

The effect of thyroxine and triiodothyronine on fatigue, depression, and memory in a hypothyroid population

Background. This study was planned at a time when important questions were being raised about the adequacy of using one hormone to treat hypothyroidism instead of two. Specifically, this trial aimed to replicate prior findings which suggested that substituting 12.5 μg of liothyronine for 50 μg of levothyroxine might improve mood, cognition, and physical symptoms. Additionally, this trial aimed to extend findings to fatigue. ^ Methods. A randomized, double-blind, two-period, crossover design was used. Hypothyroid patients stabilized on levothyroxine were invited to participate. Thirty subjects were recruited and randomized. Sequence one received their standard levothyroxine dose in one capsule and placebo in another during the first six weeks. Sequence two received their usual levothyroxine dose minus 50 μg in one capsule and 10 μg of liothyronine in another. At the end of the first six week period, subjects were crossed over. T tests were used to assess carry-over and treatment effects. ^ Results. Twenty-seven subjects completed the trial. The majority of completers had an autoimmune etiology. Mean baseline levothyroxine dose was 121 μg/d (±26.0). Subjects reported small increases in fatigue as measured by the Piper Fatigue Scale (0.9, p = 0.09) and in symptoms of depression measured by the Beck Depression Inventory-II (2.3, p = 0.16) as well as the General Health Questionnaire-30 (4.7, p = 0.14) while treated with substitution treatment. However, none of these differences was statistically significant. Measures of working memory were essentially unchanged between treatments. Thyroid stimulating hormone was about twice as high during substitution treatment (p = 0.16). Free thyroxine index was reduced by 0.7 (p < 0.001), and total serum thyroxine was reduced by 3.0 (p < 0.001) while serum triiodothyronine was increased by 20.5 (p < 0.001) on substitution treatment. ^ Conclusions. Substituting an equivalent amount of liothyronine for a portion of levothyroxine in patients with hypothyroidism does not decrease fatigue, symptoms of depression, or improve working memory. However, due to changes in serum hormone levels and small increments in fatigue and depression symptoms on substitution treatment, a question was raised about the role of T3 in the serum. ^

Bayesian joint modeling of longitudinal and survival data

The joint modeling of longitudinal and survival data is a new approach to many applications such as HIV, cancer vaccine trials and quality of life studies. There are recent developments of the methodologies with respect to each of the components of the joint model as well as statistical processes that link them together. Among these, second order polynomial random effect models and linear mixed effects models are the most commonly used for the longitudinal trajectory function. In this study, we first relax the parametric constraints for polynomial random effect models by using Dirichlet process priors, then three longitudinal markers rather than only one marker are considered in one joint model. Second, we use a linear mixed effect model for the longitudinal process in a joint model analyzing the three markers. In this research these methods were applied to the Primary Biliary Cirrhosis sequential data, which were collected from a clinical trial of primary biliary cirrhosis (PBC) of the liver. This trial was conducted between 1974 and 1984 at the Mayo Clinic. The effects of three longitudinal markers (1) Total Serum Bilirubin, (2) Serum Albumin and (3) Serum Glutamic-Oxaloacetic transaminase (SGOT) on patients' survival were investigated. Proportion of treatment effect will also be studied using the proposed joint modeling approaches. ^ Based on the results, we conclude that the proposed modeling approaches yield better fit to the data and give less biased parameter estimates for these trajectory functions than previous methods. Model fit is also improved after considering three longitudinal markers instead of one marker only. The results from analysis of proportion of treatment effects from these joint models indicate same conclusion as that from the final model of Fleming and Harrington (1991), which is Bilirubin and Albumin together has stronger impact in predicting patients' survival and as a surrogate endpoints for treatment. ^

Why does low dose morphine augment fentanyl analgesia?

Opioids dominate the field of pain management because of their ability to provide analgesia in many medical circumstances. However, side effects including respiratory depression, constipation, tolerance, physical dependence, and the risk of addiction limit their clinical utility. Fear of these side effects results in the under-treatment of acute pain. For many years, research has focused on ways to improve the therapeutic index (the ratio of desirable analgesic effects to undesirable side effects) of opioids. One strategy, combining opioid agonists that bind to different opioid receptor types, may prove successful.^ We discovered that subcutaneous co-administration of a moderately analgesic dose of the mu-opioid receptor (MOR) selective agonist fentanyl (20μg/kg) with subanalgesic doses of the less MOR-specific agonist morphine (100ng/kg-100μg/kg), augmented acute fentanyl analgesia in rats. Parallel [35S]GTPγS binding studies using naïve rat substantia gelatinosa membrane treated with fentanyl (4μM) and morphine (1nM-1pM) demonstrated a 2-fold increase in total G-protein activation. This correlation between morphine-induced augmentation of fentanyl analgesia and G-protein activation led to our proposal that interactions between MORs and DORs underlie opioid-induced augmentation. We discovered that morphine-induced augmentation of fentanyl analgesia and G-protein activity was mediated by DORs. Adding the DOR-selective antagonist naltrindole (200ng/kg, 40nM) at doses that did not alter the analgesic or G-protein activation of fentanyl, blocked increases in analgesia and G-protein activation induced by fentanyl/morphine combinations. Equivalent doses of the MOR-selective antagonist cyprodime (20ng/kg, 4nM) did not block augmentation. Substitution of the DOR-selective agonist SNC80 for morphine yielded similar results, further supporting our conclusion that interactions between MORs and DORs are responsible for morphine-induced augmentation of fentanyl analgesia and G-protein activation. Confocal microscopy of rat substantia gelatinosa showed that changes in the rate of opioid receptor internalization did not account for these effects.^ In conclusion, fentanyl analgesia augmentation by subanalgesic morphine is mediated by increased G-protein activation resulting from functional interactions between MORs and DORs, not changes in MOR internalization. Additional animal and clinical studies are needed to determine whether side effect incidence changes following opioid co-administration. If side effect incidence decreases or remains unchanged, these findings could have important implications for clinical pain treatment. ^

Secondary data analysis of a pilot study of use of an instructional video for dietary assessment

Introduction. Food frequency questionnaires (FFQ) are used study the association between dietary intake and disease. An instructional video may potentially offer a low cost, practical method of dietary assessment training for participants thereby reducing recall bias in FFQs. There is little evidence in the literature of the effect of using instructional videos on FFQ-based intake. Objective. This analysis compared the reported energy and macronutrient intake of two groups that were randomized either to watch an instructional video before completing an FFQ or to view the same instructional video after completing the same FFQ. Methods. In the parent study, a diverse group of students, faculty and staff from Houston Community College were randomized to two groups, stratified by ethnicity, and completed an FFQ. The "video before" group watched an instructional video about completing the FFQ prior to answering the FFQ. The "video after" group watched the instructional video after completing the FFQ. The two groups were compared on mean daily energy (Kcal/day), fat (g/day), protein (g/day), carbohydrate (g/day) and fiber (g/day) intakes using descriptive statistics and one-way ANOVA. Demographic, height, and weight information was collected. Dietary intakes were adjusted for total energy intake before the comparative analysis. BMI and age were ruled out as potential confounders. Results. There were no significant differences between the two groups in mean daily dietary intakes of energy, total fat, protein, carbohydrates and fiber. However, a pattern of higher energy intake and lower fiber intake was reported in the group that viewed the instructional video before completing the FFQ compared to those who viewed the video after. Discussion. Analysis of the difference between reported intake of energy and macronutrients showed an overall pattern, albeit not statistically significant, of higher intake in the video before versus the video after group. Application of instructional videos for dietary assessment may require further research to address the validity of reported dietary intakes in those who are randomized to watch an instructional video before reporting diet compared to a control groups that does not view a video.^

Proteómica de la madurez del tomate : identificación de dos estados de madurez del fruto por perfiles proteicos totales del pericarpio en RILs de tomate

Durante la madurez del fruto se producen cambios morfológicos, fisiológicos y bioquímicos provocados por la expresión regulada de diferentes genes. El objetivo de este trabajo fue verificar si la presencia de polipéptidos totales del pericarpio en los estados verde maduro (VM) y rojo maduro (RM) permite caracterizar la madurez del tomate. Se analizaron 18 líneas endocriadas recombinantes obtenidas por selección antagónica-divergente de un cruzamiento entre la cv. Caimanta (Solanum lycopersicum) y la entrada LA722 (S. pimpinellifolium), que fueron incluidas junto a la F1 como testigos experimentales. Los extractos proteicos se obtuvieron de dos muestras independientes de cada estado según el protocolo estándar y se resolvieron en SDS-PAGE. Se analizó la presencia/ausencia de bandas por genotipos y por estado, detectándose 26 en VM y 29 en RM. Algunas bandas fueron comunes entre estados, mientras que otras resultaron propias de VM o RM, respectivamente. Se calcularon las distancias de Jaccard y se realizó un análisis de conglomerados según el método UPGMA. En el dendrograma (correlación cofenética = 0,43) se distinguieron dos grandes grupos definidos por el estado de madurez. Se concluye que los perfiles proteicos del pericarpio son una herramienta postgenómica apropiada para identificar dos estados de madurez del fruto de tomate.

Biogeochemistry of sediments in the Aegean Sea

Biochemical composition of sedimentary organic matter (OM), vertical fluxes and bacterial distribution were studied at 15 stations (95-2270 m depth) in the Aegean Sea during spring and summer. Downward fluxes of labile OM were significantly higher in the northern than in the southern part and were higher in summer than in spring. Primary inputs of OM were not related to sedimentary OM concentrations, which had highest values in summer. Sedimentary chlorophyll-a concentrations were similar in the northern and southern parts. Carbohydrates, the main component of sedimentary OM, were about 1.2 times higher in the southern part than in the northern, without significant temporal changes. Total proteins were higher in summer and about double in the northern part. Sedimentary proteins appeared more dependent upon the downward flux of phytopigment than of proteins. Sedimentary OM was characterised by a relatively large fraction of soluble compounds and showed better quality in the northern part. The lack of a depth-related pattern in sedimentary OM and the similar concentrations in the two areas suggest that differences in sedimentary OM quality in the Aegean basin are dependent on system productivity; the bulk of sedimentary OM is largely conservative. Sedimentary bacterial density was about double in the northern part and higher in spring than in summer, but bacterial size was about three times higher in summer, resulting in a larger bacterial biomass in summer. Bacterial density was coupled with total and protein fluxes, indicating a rapid bacterial response to pelagic production. Bacterial biomass was significantly correlated with sedimentary protein and phytopigment concentrations, indicating a clear response to accumulation of labile OM in the sediments. In all cases bacteria accounted for <5% of the organic C and N pools. The efficiency of benthic bacteria in exploiting protein pools, estimated as amounts of protein available per unit bacterial biomass, indicates a constant ratio of about 70 µg proteins/µg C. This suggests a similar bacterial efficiency all over the area studied, unaffected by different trophic conditions.

Seasonal changes in labile organic matter composition on the continental shelf and bathyal sediments of the Cretan Sea

Bacterial abundance, biomass and cell size were studied in the oligotrophic sediments of the Cretan Sea (Eastern Mediterranean), in order to investigate their response to the seasonal varying organic matter (OM) inputs. Sediment samples were collected on a seasonal basis along a transect of seven stations (ranging from 40 to 1570 m depth) using a multiple-corer. Bacterial parameters were related to changes in chloroplastic pigment equivalents (CPE), the biochemical composition (proteins, lipids, carbohydrates) of the sedimentary organic matter and the OM flux measured at a fixed station over the deep basin (1570 m depth). The sediments of the Cretan Sea represent a nutrient depleted ecosystem characterised by a poor quality organic matter. All sedimentary organic compounds were found to vary seasonally, and changes were more evident on the continental shelf than in deeper sediments. Bacterial abundance and biomass in the sediments of the Cretan Sea (ranging from 1.02 to 4.59 * 10**8 cells/g equivalent to 8.7 and 38.7 µgC/g) were quite high and their distribution appeared to be closely related to the input of fresh organic material. Bacterial abundance and biomass were sensitive to changes in nutrient availability, which also controls the average cell size and the frequency of dividing cells. Bacterial abundance increased up to 3-fold between August '94 and February '95 in response to the increased amount of sedimentary proteins and CPE, indicating that benthic bacteria were constrained more by changes in quality rather than the quantity of the sedimentary organic material. Bacterial responses to the food inputs were clearly detectable down to 10 cm depth. The distribution of labile organic compounds in the sediments appeared to influence the vertical patterns of bacterial abundance and biomass. Cell size decreased significantly with water depth. Bacterial abundance and biomass were characterised by clear seasonal changes in response to seasonal OM pulses. The strong coupling between protein flux and bacterial biomass together with the strong bacterial dominance over the total biomass suggest that the major part of the carbon flow was channelled through the bacteria and the benthic microbial loop.

Performance data of the coral Pocillopora verrucosa and environmental conditions along the Saudi Arabian Red Sea coast as assessed during 3 expeditions in the years 2011 and 2012

Large scale patterns of ecologically relevant traits may help identify drivers of their variability and conditions beneficial or adverse to the expression of these traits. Antimicrofouling defenses in scleractinian corals regulate the establishment of the associated biofilm as well as the risks of infection. The Saudi Arabian Red Sea coast features a pronounced thermal and nutritional gradient including regions and seasons with potentially stressful conditions to corals. Assessing the patterns of antimicrofouling defenses across the Red Sea may hint at the susceptibility of corals to global change. We investigated microfouling pressure as well as the relative strength of 2 alternative antimicrofouling defenses (chemical antisettlement activity, mucus release) along the pronounced environmental gradient along the Saudi Arabian Red Sea coast in 2 successive years. Microfouling pressure was exceptionally low along most of the coast but sharply increased at the southernmost sites. Mucus release correlated with temperature. Chemical defense tended to anti-correlate with mucus release. As a result, the combined action of mucus release and chemical antimicrofouling defense seemed to warrant sufficient defense against microbes along the entire coast. In the future, however, we expect enhanced energetic strain on corals when warming and/or eutrophication lead to higher bacterial fouling pressure and a shift towards putatively more costly defense by mucus release.