Development of protein kinase activators: AMPK as a target in metabolic disorders and cancer.

AMP-activated protein kinase (AMPK) is a cellular energy sensor activated by metabolic stresses that either inhibit ATP synthesis or accelerate ATP consumption. Activation of AMPK in response to an increase in the cellular AMP:ATP ratio results in inhibition of ATP-consuming processes such as gluconeogenesis and fatty acid synthesis, while stimulating ATP-generating processes, including fatty acid oxidation. These alterations in lipid and glucose metabolism would be expected to ameliorate the pathogenesis of obesity, type 2 diabetes and other metabolic disorders. Recently, AMPK has also been identified as a potential target for cancer prevention and/or treatment. Cell growth and proliferation are energetically demanding, and AMPK may act as an "energy checkpoint" that permits growth and proliferation only when energy reserves are sufficient. Thus, activators of AMPK could have potential as novel therapeutics both for metabolic disorders and for cancer, which together constitute two of the most prevalent groups of diseases worldwide.

The effectiveness of massage for reducing pregnant women's anxiety and depression; systematic review and meta-analysis.

OBJECTIVE: To critically appraise and synthesize the best available evidence on the effectiveness of massage to reduce antenatal women's anxiety and/ or depression. DESIGN: Systematic review with meta-analysis PARTICIPANTS, INTERVENTIONS: Pregnant women over the age of 18 years who receive massage interventions. MEASUREMENTS AND FINDINGS: Eight studies were included in the review; seven were randomized controlled trials. Data were collected via pregnant women's self-reported ratings of anxiety or depression using validated tools. Meta-analysis of four studies revealed a moderate effect of massage therapy on women's depressive symptoms as measured by the Center for Epidemiologic Studies Depression Scale (CES-D) (MD = -5.95, 95%CI = -8.11 to -3.80, I2 = 0%) compared with usual care. A moderate effect of massage interventions on women's anxiety were also found based on five studies using various measures (SMD = -0.59, 95%CI = -1.06 to -0.12, I2 = 75%) when compared with usual care. However, none of the trials had a low risk of bias. KEY CONCLUSIONS: Non-pharmacologic treatments for mental health symptoms are an important option for women to use during pregnancy. As shown in meta-analysed data, massage therapy might be more effective in reducing pregnant women's anxiety and depression than usual care, although the current results may be prone to bias. Further high-quality research is required to fully evaluate the impact of massage therapy on pregnant women's mental health symptoms in the immediate and also longer term. IMPLICATIONS FOR PRACTICE: Massage therapy may be an acceptable and feasible approach for pregnant women to employ to reduce their anxiety and depressive symptoms. More research evidence examining the safety and effectiveness of massage is required before practice recommendations can be made.

A systematic review and network meta-analysis of current and investigational treatments for active ankylosing spondylitis.

OBJECTIVE: To compare the relative efficacy of current and investigational biologic and oral small molecule (OSM) treatments for active ankylosing spondylitis (AS). METHODS: A systematic literature review was conducted to identify all phase 2/3 randomized trials of interest in patients with AS. Outcomes assessed were ≥ 20% improvement in the Assessment of Spondyloarthritis International Society Criteria (ASAS20) and change from baseline in Bath Ankylosing Spondylitis Functional Index (BASFI) and C-reactive protein (CRP) at weeks 12-16. Bayesian network meta-analyses were conducted for outcomes using a random effects model. Baseline-risk adjustment was also conducted to account for differences in placebo response across studies. Surface Under the Cumulative Ranking curve (SUCRA) values are reported, reflecting the relative probability that intervention was the best of all interventions. RESULTS: The investigational agent tofacitinib 5 mg was the top-ranked treatment (SUCRA, 93%) for ASAS20 response, followed by intravenous (IV) golimumab 2 mg/kg (90%). Golimumab IV 2 mg/kg and infliximab 5 mg/kg were the top two ranked treatments for change from baseline in BASFI (golimumab IV, 81%; infliximab, 80%) and change from baseline in CRP (infliximab, 90%; golimumab IV, 82%). CONCLUSIONS: Two approved therapies (golimumab IV, infliximab) and one investigational product ranked highest for efficacy in AS. Key Points • Although golimumab IV, infliximab, and tofacitinib ranked highest for efficacy in AS, differences in efficacy between approved and investigational therapies were not statistically significant.

Mechanisms underlying the metabolic actions of galegine that contribute to weight loss in mice.

BACKGROUND AND PURPOSE: Galegine and guanidine, originally isolated from Galega officinalis, led to the development of the biguanides. The weight-reducing effects of galegine have not previously been studied and the present investigation was undertaken to determine its mechanism(s) of action. EXPERIMENTAL APPROACH: Body weight and food intake were examined in mice. Glucose uptake and acetyl-CoA carboxylase activity were studied in 3T3-L1 adipocytes and L6 myotubes and AMP activated protein kinase (AMPK) activity was examined in cell lines. The gene expression of some enzymes involved in fat metabolism was examined in 3T3-L1 adipocytes. KEY RESULTS: Galegine administered in the diet reduced body weight in mice. Pair-feeding indicated that at least part of this effect was independent of reduced food intake. In 3T3-L1 adipocytes and L6 myotubes, galegine (50 microM-3 mM) stimulated glucose uptake. Galegine (1-300 microM) also reduced isoprenaline-mediated lipolysis in 3T3-L1 adipocytes and inhibited acetyl-CoA carboxylase activity in 3T3-L1 adipocytes and L6 myotubes. Galegine (500 microM) down-regulated genes concerned with fatty acid synthesis, including fatty acid synthase and its upstream regulator SREBP. Galegine (10 microM and above) produced a concentration-dependent activation of AMP activated protein kinase (AMPK) in H4IIE rat hepatoma, HEK293 human kidney cells, 3T3-L1 adipocytes and L6 myotubes. CONCLUSIONS AND IMPLICATIONS: Activation of AMPK can explain many of the effects of galegine, including enhanced glucose uptake and inhibition of acetyl-CoA carboxylase. Inhibition of acetyl-CoA carboxylase both inhibits fatty acid synthesis and stimulates fatty acid oxidation, and this may to contribute to the in vivo effect of galegine on body weight.

Characterization of the Escherichia coli gcv operon.

The nucleotide (nt) sequence of the Escherichia coli gcvP gene was determined. The polypeptide deduced from the DNA sequence has an M(r) of 104,375 (957 amino acids). In a minicell system, gcvP encodes a polypeptide that migrates at 93.3 kDa on sodium dodecyl sulfate-polyacrylamide gels. After the coding region, there is a 39-nt sequence followed by a T-rich sequence within which transcription appears to terminate. This region is preceded by a G/C-rich sequence that could form a stable stem-loop structure once transcribed, and is characteristic of Rho-independent transcription terminators. A Northern analysis identified an approx. 4700-nt RNA molecule, large enough to encode the T-, H-and P-proteins of the glycine cleavage enzyme complex. Analyses of gcvP::lacZ fusions with and without stop codons in gcvT, the first gene in the operon, confirmed gcvT, gcvH and gcvP lie in an operon. RNA slot blot analyses indicated that induction of gcv by glycine, and PurR-mediated repression of gcv occur at the level of transcription.

Soluble fibrin augments platelet/tumor cell adherence in vitro and in vivo, and enhances experimental metastasis.

There is considerable evidence for a relationship between hemostasis and malignancy. Since platelet adhesion to tumor cells has been implicated in the metastatic process and plasma levels of fibrinogen (Fg) and soluble fibrin (sFn) monomer are increased in cancer, we hypothesized that these molecules might enhance tumor-platelet interaction. We therefore studied binding of sFn monomer to tumor cells in a static microplate adhesion assay and determined the effect of pre-treating tumor cells with sFn on tumor cell-induced thrombocytopenia and experimental metastasis. Soluble fibrin (produced by adding thrombin to FXIII- and plasminogen-free Fg in the presence of Gly-Pro-Arg-Pro-amide (GPRP-NH2) significantly increased platelet adherence to tumor cells. This effect was primarily mediated by the integrins alphaIIb beta3 on the platelet and CD 54 (ICAM-1) on the tumor cells. Platelets adhered to untreated A375 cells (28 +/- 8 platelets/tumor cell) and this was not significantly affected by pre-treatment of the tumor cells with fibrinogen or GPRP-NH2. Although thrombin treatment increased adherence, pre-incubation of the tumor cells with sFn resulted in a further increase in platelet binding to tumor cells. In contrast to untreated tumor cells, intravenous injection of sFn-treated A 375 cells reduced the platelet count in anticoagulated mice, supporting the in vitro finding that sFn enhanced tumor cell-platelet adherence. In a more aggressive model of experimental metastasis, treating tumor cells with sFn enhanced lung seeding by 65% compared to untreated cells. Extrapolation of our data to the clinical situation suggests that coagulation activation, and subsequent increase in circulating Fn monomer, may enhance platelet adhesion to circulating tumor cells and thereby facilitate metastatic spread.

Choroidal neovascularization with granulomatous inflammation in ocular histoplasmosis syndrome.

PURPOSE: To report pathologic examination of an excised choroidal neovascular membrane in a patient with ocular histoplasmosis syndrome that demonstrated granulomatous inflammation. METHOD: Case report. A 50-year-old woman with sudden vision loss in her left eye demonstrated clinical and fluorescein angiographic findings characteristic of choroidal neovascularization secondary to ocular histoplasmosis syndrome. RESULTS: Histopathologic examination of the surgically excised choroidal neovascular membrane disclosed granulomatous inflammation. CONCLUSIONS: This case suggests an important role of mononuclear phagocytic cells as primary mediators of angiogenesis or modifiers of choroidal neovascularization. This association of choroidal neovascularization with granulomatous inflammation did not respond to treatment with systemic corticosteroids.

The application of high-throughput screening to novel lead discovery.

The ability to discover new lead compounds for novel therapeutic targets is a pivotal step in drug discovery programmes. High-throughput screening (HTS) utilises a number of platforms for the rapid screening of novel targets to accelerate this process. Key issues in HTS include assay configuration and the ability of a high-throughput screen to predict drug-target interactions accurately. This review highlights a number of issues in the HTS process and describes three key target areas that are likely to be sources of novel, therapeutically important drugs. Particular emphasis is placed on the mechanistic basis of drug-target interactions that are of prime importance in the design of HTS approaches. Critical aspects of information management related to HTS are summarised.

Effect of passive range of motion on intracranial pressure in neurosurgical patients.

PURPOSE: A prospective patient study was done to evaluate the effect of passive range of motion (PROM) on intracranial pressure (ICP) and cerebral perfusion pressure (CPP) in neurosurgical patients. MATERIALS AND METHODS: Twelve adult patients admitted to the neurological-neurosurgical intensive care unit of a community teaching hospital were enrolled in the study. The study patients all required ICP monitoring and they underwent a total of 20 PROM sessions. Six patients (10 PROM sessions) were mechanically ventilated, and six patients (10 PROM sessions) were breathing spontaneously. Mean arterial pressure (MAP), heart rate (HR), respiratory rate (RR), and ICP were measured at baseline and at 1-minute intervals during the physical therapy session. CPP was calculated as the difference between MAP and ICP. RESULTS: No significant changes were detected in MAP, ICP, HR, or RR during the study period. Calculated CPP remained unchanged. Mean duration of PROM intervention was 7 +/- 1 minute. Mean Glasgow Coma Scale (GCS) for mechanically ventilated patients was 7 +/- 1.3 and for spontaneously breathing patients 13 +/- 0.8. CONCLUSION: PROM results in no significant changes in ICP or CPP in stable, neurosurgical patients in the absence of intracranial hypertension.

Personality-dependent dispersal: characterization, ontogeny and consequences for spatially structured populations.

Dispersal is one of the most fundamental components of ecology, and affects processes as diverse as population growth, metapopulation dynamics, gene flow and adaptation. Although the act of moving from one habitat to another entails major costs to the disperser, empirical and theoretical studies suggest that these costs can be reduced by having morphological, physiological or behavioural specializations for dispersal. A few recent studies on different systems showed that individuals exhibit personality-dependent dispersal, meaning that dispersal tendency is associated with boldness, sociability or aggressiveness. Indeed, in several species, dispersers not only develop behavioural differences at the onset of dispersal, but display these behavioural characteristics through their life cycle. While personality-dependent dispersal has been demonstrated in only a few species, we believe that it is a widespread phenomenon with important ecological consequences. Here, we review the evidence for behavioural differences between dispersers and residents, to what extent they constitute personalities. We also examine how a link between personality traits and dispersal behaviours can be produced and how personality-dependent dispersal affects the dynamics of metapopulations and biological invasions. Finally, we suggest future research directions for population biologists, behavioural ecologists and conservation biologists such as how the direction and the strength of the relationship between personality traits and dispersal vary with ecological contexts.

A penetrating inferior vena caval filter...

INTRODUCTION: The concept of vena caval interruption was first described by Armand Trousseau in 1865 [HANN CL. STREIFF MB. The role of vena caval filters in the management of venous thromboembolism. Blood Rev 2005;19(4):179-202]. Methods described have included surgical ligation of the inferior vena cava (IVC) to the development of percutaneous inferior vena caval filters which have been, up until recently, permanently placed in the vessel. These devices are not without risk. We describe a case of penetration of the duodenum by a standard stainless steel Greenfield vena caval filter. REPORT: An 83 year old woman presented with vague epigastric discomfort. She was noted to have intermittent globally deranged liver function tests and was investigated accordingly. Investigations included an endoscopic retrograde cholangio-pancreatography (ERCP) which revealed an inferior vena cava filter penetrating the wall of the small bowel. CONCLUSION: We describe this unique presentation of a penetrating vena caval filter and suggest that a clinician may be inclined to consider a simple endoscopy on occasions when this complication/diagnosis is considered.

Bacteriophage lambda DNA packaging: a mutant terminase that is independent of integration host factor.

Lambda+ is able to grow in Escherichia coli cells lacking integration host factor (IHF), producing a burst of approximately 25% that produced in IHF+ cells. In vitro, however, we find that the lambda DNA packaging enzyme terminase is strongly dependent on IHF in both cos cleavage reactions and DNA packaging reactions. The cos59 mutation renders lambda dependent on IHF in vivo. The cos59 mutation is a deletion of 3 base pairs at the XmnI site in the cohesive end site (cos) of lambda. Variants of lambda cos59 that were able to grow in the absence of IHF were isolated and found to carry a mutation, called ms1, in the Nu1 gene, which codes for the small subunit of terminase. The Nu1ms1 mutation results in a change of the 40th amino acid of the Nu1 gene product from leucine to phenylalanine. The Nu1ms1 terminase was independent of IHF in packaging reactions in vitro. The results indicate that the mutation either renders terminase: (1) able to utilize some host protein other than IHF, or (2) totally independent of host factors.

Depression and the accessibility of memories. A longitudinal study.

A group of depressed patients and a group of normal subjects were assessed on two occasions. On each they were required to recall past real life experiences associated with a series of stimulus words. Subjects were subsequently required to rate each experience for happiness and pleasantness. The depressives showed an increased probability of recalling sad memories; within this group there was a significant relationship between severity of depressed mood and the percentage of sad memories recalled. The extent of reduction in depressed mood from occasion 1 to occasion 2 was significantly related to the increase in accessibility of happy memories and the decrease in recall of sad memories. For those patients who improved between the two occasions there was a significant reduction in the probability of recall of a very unhappy memory. The results extend the previous findings on the effect of mood state on the accessibility of memories of differing affective tone. The findings are discussed in terms of context specific encoding and retrieval.

Multiple regions in the Rous sarcoma virus src gene intron act in cis to affect the accumulation of unspliced RNA.

Retrovirus replication requires the production of both spliced and unspliced viral RNA from a single RNA transcript. In contrast, cellular RNA precursors with introns almost completely spliced. The cis elements and virus-coded trans factors which determine the extent to which Rous sarcoma virus RNA is spliced to src mRNA were investigated by transfecting chicken embryo fibroblasts with cloned wild-type and mutant DNA followed by the analysis of viral RNA. Two cis-acting regions important in the negative control of splicing were detected. Cell cultures transfected with a clone deleted in 80% of the src intron (nucleotide 1149 to nucleotide 6574) demonstrated only a 2-fold reduction in the ratio of unspliced to spliced RNA relative to the wild-type clone, whereas cultures transfected with clones which were further deleted in the gag gene region (between nucleotide 630 and nucleotide 5258) demonstrated an approximate 20-fold reduction. Cell cultures which were transfected with clones deleted only between nucleotides 543 and 1806 demonstrated only a three- to fourfold reduction in the unspliced-to-spliced RNA ratio, suggesting that at least one other region of the src intron can partially compensate for the deletion of the gag region. Both regions appeared to act in cis on the distribution of unspliced and spliced RNA since the ratios were not altered by cotransfection with helper virus DNA.

Speech rate as a measure of short-term variation in depression.

Experimental investigation of the state of depression is at present hindered by the lack of simple, objective, repeatable measures of components of the state. Covariation between self-report measures of affect and simple measure of speech rate was investigated in a series of studies of both clinical and normal subjects, using single-case and group designs. Slowing of speech was significantly and substantially correlated with self-reports of increased unpleasant affect in certain situations. This suggests that the speech measure may usefully reflect a behavioural component of a state of retarded-type depression.

The arterial supply to lymph nodes in sheep.

The aim of this study was to describe how lymph nodes in sheep receive their blood supply, and to provide some information on the pathways followed by arteries and their branches within the nodes. It was found that considerable variation exists in the arterial supply to lymph nodes in sheep. Each node may receive branches from one, two or three major arteries; the nodes may be up to 10-15 cm from these arteries, but some arteries actually pass through lymph nodes. Most branches enter at a hilus, which varies in shape, size and location, but some penetrate the capsule directly. Within the node, arteries may either proceed directly towards the cortex or extend longitudinally through the medulla; in each case branches are given off to supply the medulla and, in greater numbers, the cortex. Some arteries then leave the node through the capsule and either supply perinodal fat and/or more distal structures, or join other arteries. Anastomoses may also occur before the arteries enter a node, or within the node or at its surface. These direct anastomoses and arterial circles could play a part in maintaining the blood supply to a node when it is being bombarded with microorganisms and inflammatory products from an area of infection.

Purification of insect vitellogenin and vitellin by gel-immobilized ferric chelate.

Vitellogenin and vitellin of Manduca sexta and some other insect species were purified by immobilized metal ion affinity chromatography. Ferric ion was chosen as the immobilized metal ion. Agarose-bound carboxymethylpicolylamine was used as the chelating adsorbent for the ferric ion. Vitellogenin and vitellin, both phosphorylated lipoproteins, were shown to bind specifically to the iron. The general applicability of immobilized ferric ion affinity chromatography for the purification of insect vitellogenin and vitellin is suggested.