Impacts of increased ocean temperatures on a low-latitude coral reef fish - Processes related to oxygen uptake and delivery.

Increasing temperatures are expected to significantly affect the physiological performance of ectotherms, particularly in tropical locations. The shape of an organism's thermal reaction norm can provide important information on its capacity to persist under climate change scenarios; however, difficulty lies in choosing a measurable trait that best depicts physiological performance. This study investigated the effects of elevated temperatures on processes related to oxygen uptake and delivery, including oxygen consumption, haematology, and tissue health for a low-latitude population of coral reef damselfish. Acanthochromis polyacanthus were collected from the Torres Strait (10°31-46'S, 142°20-35'E) and maintained at current average ocean temperatures (+0 °C; seasonally cycling), + 1.5 °C and + 3 °C higher than present day temperatures for 10 months. Aerobic performance indicated a limit to metabolic function at + 3 °C (33 °C), following an increase in aerobic capacity at + 1.5 °C (31.5 °C). Neither haematological parameters nor gill morphology showed the same improvement in performance at + 1.5 °C. Gill histopathology provided the first indicator of a decline in organism health, which corresponded with mortality observations from previous research. Findings from this study suggest thermal specialisation in this low-latitude population as well as variation in thermal sensitivity, depending on the physiological trait.

Treatment with methylnaltrexone is associated with increased survival in patients with advanced cancer.

BACKGROUND: Methylnaltrexone (MNTX), a peripherally acting µ-opioid receptor (MOR) antagonist, is FDA-approved for treatment of opioid-induced constipation (OIC). Preclinical data suggest that MOR activation can play a role in cancer progression and can be a target for anticancer therapy. PATIENTS AND METHODS: Pooled data from advanced end-stage cancer patients with OIC, despite laxatives, treated in two randomized (phase III and IV), placebo-controlled trials with MNTX were analyzed for overall survival (OS) in an unplanned post hoc analysis. MNTX or placebo was given subcutaneously during the double-blinded phase, which was followed by the open-label phase, allowing MNTX treatment irrespective of initial randomization. RESULTS: In two randomized, controlled trials, 229 cancer patients were randomized to MNTX (117, 51%) or placebo (112, 49%). Distribution of patients' characteristics and major tumor types did not significantly differ between arms. Treatment with MNTX compared with placebo [76 days, 95% confidence interval (CI) 43-109 versus 56 days, 95% CI 43-69; P = 0.033] and response (laxation) to treatment compared with no response (118 days, 95% CI 59-177 versus 55 days, 95% CI 40-70; P < 0.001) had a longer median OS, despite 56 (50%) of 112 patients ultimately crossing over from placebo to MNTX. Multivariable analysis demonstrated that response to therapy [hazard ratio (HR) 0.47, 95% CI 0.29-0.76; P = 0.002) and albumin ≥3.5 (HR 0.46, 95% CI 0.30-0.69; P < 0.001) were independent prognostic factors for increased OS. Of interest, there was no difference in OS between MNTX and placebo in 134 patients with advanced illness other than cancer treated in these randomized studies (P = 0.88). CONCLUSION: This unplanned post hoc analysis of two randomized trials demonstrates that treatment with MNTX and, even more so, response to MNTX are associated with increased OS, which supports the preclinical hypothesis that MOR can play a role in cancer progression. Targeting MOR with MNTX warrants further investigation in cancer therapy. CLINICAL TRIALS NUMBER: NCT00401362, NCT00672477.

Breath carbon stable isotope ratios identify changes in energy balance and substrate utilization in humans.

Rapid detection of shifts in substrate utilization and energy balance would provide a compelling biofeedback tool for individuals attempting weight loss. As a proof of concept, we tested whether the natural abundance of exhaled carbon stable isotope ratios (breath δ(13)C) reflects shifts between negative and positive energy balance. Volunteers (n=5) consumed a 40% energy-restricted diet for 6 days followed by 50% excess on day 7. Breath was sampled immediately before and 1 h and 2 h after breakfast, lunch and dinner. Exhaled breath δ(13)C values were measured by cavity ring-down spectroscopy. Using repeated measures analysis of variance (ANOVA) followed by Dunnett's contrasts, pre-breakfast breath values on days 2-6 were compared with day 1, and postprandial day 7 time points were compared with pre-breakfast day 7. Energy restriction diminished pre-breakfast breath δ(13)C by day 3 (P<0.05). On day 7, increased energy intake was first detected immediately before dinner (-23.8±0.6 vs -21.9±0.7‰, P=0.002 (means±s.d.)), and breath δ(13)C remained elevated at least 2 h post dinner. In conclusion, when shifting between negative and positive energy balance, breath δ(13)C showed anticipated isotopic changes. Although additional research is needed to determine specificity and repeatability, this method may provide a biomarker for marked increases in caloric intake.

Effect of clothing weight on body weight.

BACKGROUND: In clinical settings, it is common to measure weight of clothed patients and estimate a correction for the weight of clothing, but we can find no papers in the medical literature regarding the variability in clothing weight of adults with weather, season and gender. METHODS: Fifty adults (35 women) were weighed four times during a 12-month period with and without clothing. Clothing weights were determined and regressed against minimum, maximum and average daily outdoor temperature. RESULTS: The average clothing weight (±s.d.) throughout the year was significantly greater in men than in women (1.2±0.3 vs 0.8±0.3 kg, P<0.0001). The average within-person minimum and the average within-person maximum clothing weights across the year were 0.9±0.2 and 1.5±0.4 kg for men, and 0.5±0.2 and 1.1±0.4 kg for women, respectively. The within-person s.d. in clothing weight was 0.3 kg for both men and women. Over the 55 °C range in the lowest to the highest outdoor temperatures, the regressions predicted a maximal change in clothing weight of only 0.4 kg in women and 0.6 kg in men. CONCLUSION: The clothing weight of men is significantly greater than that of women, but there is little variability throughout the year. Therefore, a clothing adjustment of approximately 0.8 kg for women and 1.2 kg for men is appropriate regardless of outdoor temperature.

Vascular endothelial cells maintained in the absence of fibroblast growth factor undergo structural and functional alterations that are incompatible with their in vivo differentiated properties.

Vascular endothelial cells cultured in the presence of fibroblast growth factor (FGF) adopt at confluence a morphological appearance similar to that of the vascular endothelium in vivo. Similarly, their apical cell surface is, as in vivo, nonthrombogenic. In contrast, when the cultures are maintained in the absence of FGF, the cells undergo within two to three passages structural and functional alterations that are incompatible with their in vivo morphological appearance and physiological function. Cultures maintained in the absence of FGF no longer adopt, upon reaching confluence, the configuration of a monolayer composed of small closely apposed and nonoverlapping, cuboidal cells. Instead, confluent cultures deprived of FGF consist of large, overlapping cells which have lost the polarity of cell surface characteristic of the vascular endothelium. The apical cell surface becomes thrombogenic, as reflected by its ability to bind platelets, whereas fibronectin, which at confluence is normally associated only with the basal cell surface, can be found both on top of and underneath the cell layer. Among other changes, both sparse and confluent cultures maintained in the absence of FGF showed a greatly increased production of fibronectin. CSP-60, a cell surface protein whose appearance is correlative with the adoption of a cell monolayer configuration, can no longer be detected in cultures maintained in the absence of FGF. Overlapping endothelial cells maintained in the absence of FGF can also no longer function as a protective barrier against the uptake of ligands such as low density lipoprotein. Exposure of the culture to FGF induces a restoration of the normal endothelial characteristics concomitant with the adoption of a flattened cell monolayer morphology. These results demonstrate that, in addition to being a mitogen. FGF is involved in controlling the differentiation and phenotypic expression of the vascular endothelium. This is reflected by its effect on the morphological appearance, polarity of cell surfaces, platelet binding capacity, and barrier function of the vascular endothelium.

Glucocorticoid inhibition of vascular smooth muscle cell proliferation: influence of homologous extracellular matrix and serum mitogens.

We examined the influence of glucocorticoid hormones on the proliferation of cultured adult bovine aortic smooth muscle cells (BASM) using both primary mass cultures and a cloned strain. Cloned BASM cells maintained on plastic culture dishes were inhibited by approximately 40% by dexamethasone treatment but showed no inhibition when grown of homologous extracellular matrix (ECM) coated dishes. Dexamethasone inhibited growth of primary cultures by 73% on plastic and by 45% on ECM. The inhibitory effect was specific for the glucocorticoids, dexamethasone, corticosterone, and cortisol and was not observed with progesterone, aldosterone, estradiol or 17-alpha OH progesterone. In cloned cells, the abolition of glucocorticoid inhibition by ECM was independent of seeding density and serum concentration. The inhibition on plastic was dependent on serum concentrations greater than 1% and resulted in both a slow rate of proliferation and a lower saturation density. A specific subset of peptides detected on two-dimensional gels was induced by glucocorticoids under growth inhibitory conditions but was not induced when the cells were grown on ECM. Primary cultures grown on ECM and exposed to Dulbecco's modified Eagle's Medium (DME) containing high density lipoprotein and transferrin grew at 40% of the rate observed for cultures exposed to DME with 10% serum. Both conditions showed growth inhibition of 70% in the presence of dexamethasone. The addition of epidermal and platelet-derived growth factors in DME containing high density lipoprotein and transferrin to cells grown on ECM resulted in growth rates comparable to that observed with cultures exposed to 10% serum and were inhibited 45% by dexamethasone. These results suggest that glucocorticoids inhibit smooth muscle proliferation by decreasing the sensitivity of the cells to mitogenic stimulation by high density lipoprotein when the cells are maintained on a homologous substrate.

Glucocorticoids stimulate collagen and noncollagen protein synthesis in cultured vascular smooth muscle cells.

The effect of glucocorticoids on collagen synthesis was examined in cultured bovine aortic smooth muscle (BASM) cells. BASM cells treated with 0.1 microM dexamethasone during their proliferative phase (11 d) were labeled with [3H]proline for 24 h, and the acid-precipitable material was incubated with bacterial collagenase. Dexamethasone produced an approximate twofold increase in the incorporation of proline into collagenase-digestible protein (CDP) and noncollagen protein (NCP) in the cell layer and medium. The stimulation was present in both primary mass cultures and cloned BASM. An increase in CDP and NCP was detected at 0.1 nM, while maximal stimulation occurred at 0.1 microM. Only cells exposed to dexamethasone during their log phase of growth (1-6 d after plating) showed the increase in CDP and NCP when labeled 11 d after plating. The stimulatory effect was observed in BASM cells treated with the natural bovine glucocorticoid, cortisol, dexamethasone, and testosterone, but was absent in cells treated with aldosterone, corticosterone, cholesterol, 17 beta-estradiol, and progesterone. The increase in CDP and NCP was absent in cells treated with the inactive glucocorticoid, epicortisol, and totally abolished by the antagonist, 17 alpha-hydroxyprogesterone, suggesting that the response was mediated by specific cytoplasmic glucocorticoid receptors. Dexamethasone-treated BASM cells showed a 4.5-fold increase in the specific activity of intracellular proline, which was the result of a twofold increase in the uptake of proline and depletion of the total proline pool. After normalizing for specific activity, dexamethasone produced a 2.4- and 2.8-fold increase in the rate of collagen and NCP synthesis, respectively. Cells treated with dexamethasone secreted 1.7-fold more collagen protein in 24 h compared to control cultures. The BASM cells secreted 70% Type I and 30% Type III collagen into the media as assessed by two-dimensional gel electrophoresis. The ratio of these two types was not altered by dexamethasone. The results of the present study demonstrate that glucocorticoids can act directly on vascular smooth muscle cells to increase the synthesis and secretion of collagen and NCP.

Neuroendocrine carcinoma of the liver and gallbladder in a cow.

A 15-year-old Limousin-cross cow was presented for examination with neurological signs and serum biochemical changes consistent with liver disease. Necropsy revealed enlargement of the liver with multifocal firm, depressed, pale, circumscribed lesions throughout the parenchyma. Within the gallbladder there were exophytic and villiform mucosal masses. Microscopically, hepatic structure was displaced by neoplastic cells forming trabeculae, nests and rosettes. There was transmural infiltration of the gallbladder by similar cells. The histological pattern of growth of the neoplastic cells, the presence of silver-stained cytoplasmic granules within these cells and the immunohistochemical demonstration of chromogranin A supported the diagnosis of neuroendocrine carcinoma. Bovine liver and gallbladder neuroendocrine carcinomas are rare and this is the first detailed documentation of the disease in the United Kingdom.

Histological observations of bovine tuberculosis in lung and lymph node tissues from British deer.

Deer are recognized as hosts of Mycobacterium bovis and assessing the role of wild cervids in perpetuating tuberculosis among cattle has motivated extensive research on several continents. In this paper, the histopathology of lymph node and lung tuberculous granulomas in M. bovis positive British deer is presented. The overall aim was to seek further insights into the potential for onward transmission from infected deer to other species, including cattle. Samples were obtained from an extensive survey of wild mammals in South-West England and from statutory tuberculosis surveillance. M. bovis culture-positive samples were characterised microscopically as to their stage of lesion advancement, number of acid-fast bacilli and granuloma encapsulation. Seventy percent of the deer developed granulomas containing far greater numbers of M. bovis bacilli than typically reported in cattle. Red and fallow deer had the largest number of poorly encapsulated granulomas often containing many hundreds of bacilli. The results are consistent with infected wild British deer being a potential source of environmental contamination and onward transmission to other species. However, further work on levels of bacillary shedding is required before this can be confirmed.

Effects of probiotics on body weight, body mass index, fat mass and fat percentage in subjects with overweight or obesity: a systematic review and meta-analysis of randomized controlled trials.

A systematic review and meta-analysis of randomized controlled trials was conducted to examine the effects of probiotic supplementation on body weight, body mass index (BMI), fat mass and fat percentage in subjects with overweight (BMI 25-29.9 kg m-2 ) or obesity (BMI ≥30 kg m-2 ). MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials were searched for studies published between 1946 and September 2016. A meta-analysis, using a random effects model, was performed to calculate the weighted mean difference between the intervention and control groups. Of 800 studies identified through the literature search, 15 were finally included. The studies comprised a total of 957 subjects (63% women), with the mean BMI being 27.6 kg m-2 and the duration of the interventions ranging from 3 to 12 weeks. Administration of probiotics resulted in a significantly larger reduction in body weight (weighted mean difference [95% confidence interval]; -0.60 [-1.19, -0.01] kg, I2  = 49%), BMI (-0.27 [-0.45, -0.08] kg m-2 , I2  = 57%) and fat percentage (-0.60 [-1.20, -0.01] %, I2  = 19%), compared with placebo; however, the effect sizes were small. The effect of probiotics on fat mass was non-significant (-0.42 [-1.08, 0.23] kg, I2  = 84%).

Renal mechanism of action of rat atrial natriuretic factor.

There has been conflict as to whether crude extracts of atrial natriuretic factor increase renal solute excretion by a hemodynamic mechanism or by direct inhibition of tubular transport. To investigate this issue, seven rats were studied during a euvolemic control period and following continuous administration of pure, synthetic 24 amino acid atrial natriuretic factor. A 10-25-fold increase in urinary sodium and chloride excretion occurred with a brisk kaliuresis but little bicarbonaturia. Atrial natriuretic factor caused whole kidney glomerular filtration rate to increase from 1.17 +/- 0.04 to 1.52 +/- 0.07 ml/min (P less than 0.005). A parallel increase in single nephron glomerular filtration rate, from 34 +/- 1 to 44 +/- 2 nl/min (P less than 0.001), and from 26 +/- 1 to 37 +/- 2 nl/min (P less than 0.005) was measured at the end-proximal and early distal nephron sites, respectively. Appropriate for the higher flows were an increase in absolute proximal and loop reabsorptive rates for bicarbonate, chloride, and water, with a slight decrease in fractional solute and volume reabsorption in proximal and loop segments. To exclude the possibility that atrial natriuretic factor increased filtration rate only in anesthetized animals, eight unanesthetized rats were studied. Glomerular filtration rate increased by 45%, from 2.04 +/- 0.17 to 2.97 +/- 0.27 ml/min (P less than 0.005) without significant change in renal plasma flow, as reflected by 14C-para-aminohippurate clearance (5.4 +/- 0.5-5.6 +/- 0.9 ml/min). The clearance and micropuncture data did not preclude changes in relative blood flow distribution to or in transport by deep nephron segments. In conclusion, atrial natriuretic factor appears to increase renal solute excretion predominantly by a hemodynamic mechanism without directly inhibiting superficial tubular transport.

Evaluation of the antiinflammatory and phospholipase-inhibitory activity of calpactin II/lipocortin I.

We have examined the ability of a highly purified 38-kD phospholipase-inhibitory protein (p38) isolated from human placental membranes that is also a preferred substrate for the epidermal growth factor-urogastrone (EGF-URO) receptor/kinase, to block the release of arachidonate from zymosan-stimulated murine peritoneal macrophages in vitro and to exhibit antiinflammatory activity in a carrageenin rat paw edema test in vivo. The ability of glucocorticoids to increase the amounts of this protein in macrophage cultures was also examined. p38 represents the naturally occurring, intact, NH2-terminally blocked human placental form of the protein termed calpactin II (or lipocortin I), for which partial amino acid sequence data and a complete amino acid sequence deduced from cDNA analysis have been reported. Our data demonstrated that, whereas p38 was an effective inhibitor of pancreatic phospholipase A2 in vitro, it was unable to inhibit either the release of arachidonate from cultured zymosan-stimulated mouse peritoneal macrophages or inflammation in a rat paw edema test. At comparatively high protein concentrations, p38 enhanced either arachidonate release from intact macrophages in vitro (0.5-10 micrograms/ml) or carrageenin-induced paw swelling in vivo (2.5 or 25 micrograms per injection). Furthermore, we were unable to detect induced amounts of p38 in cultures of glucocorticoid-treated peritoneal macrophages obtained from either mice or rats. Our data indicate that the antiphospholipase activity of p38 in vitro and the ability of p38 to serve as a receptor/kinase substrate may in no way relate to the putative ability of the protein to modify eicosanoid release from macrophages in vivo, so as to modulate the inflammatory process. Our data also raise the possibility that p38 (calpactin II) may not be a true representative of the lipocortin family of glucocorticoid-inducible antiinflammatory proteins, despite its ability to inhibit phospholipase A2 in vitro.

Scanning optical pyrometer for measuring temperatures in hollow cathodes.

Life-limiting processes in hollow cathodes are determined largely by the temperature of the electron emitter. To support cathode life assessment, a noncontact temperature measurement technique which employs a stepper motor-driven fiber optic probe was developed. The probe is driven inside the hollow cathode and collects light radiated by the hot interior surface of the emitter. Ratio pyrometry is used to determine the axial temperature profile. Thermocouples on the orifice plate provide measurements of the external temperature during cathode operation and are used to calibrate the pyrometer system in situ with a small oven enclosing the externally heated cathode. The diagnostic method and initial measurements of the temperature distribution in a hollow cathode are discussed.

Risk factors for estrogen receptor-rich and estrogen receptor-poor breast cancers.

A population-based case-control study was conducted in King County, WA, to investigate whether risk factors for estrogen receptor (ER)-rich and ER-poor breast cancers differ. Responses to interviews with 329 women with breast cancer who were between 25 and 54 years of age at the time of diagnosis were compared to responses of 332 women of similar age who were selected from female residents of King County by random digit dialing. Of the 329 interviewed cases, 143 had ER-rich tumors, 97 had ER-poor tumors, and 89 had tumors that were not assayed for receptors. The relative risks of ER-rich and ER-poor breast cancers were similar with respect to late menarche, single marital status, history of extended lactation, menopause before age 40, history of benign breast disease, positive family history of breast cancer, obesity, and history of oral contraceptive and noncontraceptive estrogen use. Late age at first full-term pregnancy was a risk factor for ER-rich breast cancer but not for ER-poor breast cancer. This finding suggests that different causal mechanisms operate for these two types of breast cancer and supports the hypothesis that an early first birth protects against breast cancer by reducing the level of ERs in the mammary epithelial cells from which carcinomas develop.

Developmental transitions between chromatin-bound and soluble RNA polymerase subspecies in the soybean hypocotyl.

RNA polymerase enzymes isolated from soybean hypocotyl tissue during successive developmental stages (2-8 days old) have been fractionated by Sephadex column isoelectric focusing. Both the enzymes bound to chromatin and those enzymes free in the soluble phase were investigated during development with respect to their distribution within these two pools. All observed activites were classified according to their alpha-amanitin sensitivity and isoelectric points. Two Class I subspecies (Ia, Ib) and two Class III subspecies (IIIa, IIIb) were continually present bound to chromatin throughout the developmental sequences except the IIb form which was absent at the latest stage. However, a great multiplicity (9 total) of Class II activities (totally inhibited by alpha-amanitin) were observed to be bound to chromatin at the 2nd day stage. These forms were first released from the chromatin complex and recovered in a soluble pool (4th day stage). Subsequent hypocotyl development was accompanied by the gradual disappearance of these Class II subspecies from this pool (6th day) until only two soluble species and one chromatin-bound Class II activity remained (8th day). These observations indicate that the early development of this tissue is accompanied by a dramatic alteration in the conplexity of chromatin-bound RNA polymerase subspecies. Such events may in part determine the domain of RNA secies synthesized at successive developmental stages.

Synthesis of growth hormone, prolactin, and proopiomelanocortin by intact adult ovine pituitary tissue in vitro.

Synthesis of GH, PRL, and proopiomelanocortin (POMC) by the adult ovine pituitary was examined by culturing intact tissue explants in vitro and analyzing newly synthesized proteins by two-dimensional gel electrophoresis. Spots on autoradiographs of two-dimensional gels were identified by comigration with known standards, by analysis of tryptic peptides, or both. GH and PRL are the predominant proteins synthesized in the adult ovine pituitary, but their syntheses could not be detected in the neurointermediate lobe of the pituitary, the cerebral cortex, hypothalamus, mammillary body, or placenta. As quantified by cell-free translation in rabbit reticulocyte lysate, 14% of female ovine pituitary mRNA encodes pre-GH and 30% encodes pre-PRL. As determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, ovine pre-GH and pre-PRL have molecular weights of 24,500 and 26,000, respectively, indicating they contain leader peptides of 26-30 amino acids, as found in bovine, rat, and human pre-GH and pre-PRL. POMC, the precursor to corticotropin, endorphin, and other peptides, was synthesized primarily in the neurointermediate lobe of the pituitary, but synthesis was also detected in the anterior lobe. POMC synthesis was not detected in the hypothalamus, cerebral cortex, mammillary body, or placenta.

Differential regulation by glucocorticoids of proopiomelanocortin mRNA levels in the anterior and intermediate lobes of the rat pituitary.

Glucocorticoids negatively regulate de novo synthesis of proopiomelanocortin (POMC) and the steady state POMC mRNA levels in the rat pituitary anterior lobe. The POMC system of the intermediate lobe is far less sensitive to alterations in circulating glucocorticoid levels. Possible mechanisms for this differential regulation of POMC gene expression in the two tissues are discussed.