β-Glucuronidase activation of latent aggression-promoting cues in mouse bladder urine

The present study was designed to investigate the chemical properties of the aggression-promoting cues present in bladder urine of male mice. The results of the first experiment confirmed earlier work by demonstrating the presence of an aggression-promoting chemosignal in bladder urine. In Experiment 2, behavioral assays were separately performed on the organic and aqueous layers of bladder urine obtained by repeated dichloromethane extractions. Only the combined organic layers of the initial three extractions demonstrated behavioral activity. A fourth extraction showed no behavioral activity for both organic and aqueous layers. However, the findings of Experiment 3 showed that incubation of the aqueous layer from the third CH₂Cl₂ extraction in β-glucuronidase can free additional aggression-promoting cues into a subsequent CH₂Cl₂ extraction. It is concluded that two forms of the aggression-promoting chemosignal are present in bladder urine. One is lipophilic and behaviorally active, whereas the other is conjugated, possessing latent chemosignal properties.     

Intestinal receptor for heat-stable enterotoxin of Escherichia coli is tightly coupled to a novel form of particulate guanylate cyclase.

A novel form of particulate guanylate cyclase tightly coupled by cytoskeletal components to receptors for heat-stable enterotoxin (ST) produced by Escherichia coli can be found in membranes from rat intestinal mucosa. Intestinal particulate guanylate cyclase was resistant to solubilization with detergent alone, with only 30% of the total enzyme activity being extracted with Lubrol-PX. Under similar conditions, 70% of this enzyme was solubilized from rat lung membranes. The addition of high concentrations of sodium chloride to the extraction buffer resulted in greater solubilization of particulate guanylate cyclase from intestinal membranes. Although extraction of intestinal membranes with detergent and salt resulted in greater solubilization of guanylate cyclase, a small fraction of the enzyme activity remained associated with the particulate fraction. This activity was completely resistant to solubilization with a variety of detergents and chaotropes. Particulate guanylate cyclase and the ST receptor solubilized by detergent retained their abilities to produce cyclic GMP and bind ST, respectively. However, ST failed to activate particulate guanylate cyclase in detergent extracts. In contrast, guanylate cyclase resistant to solubilization remained functional and coupled to the ST receptor since enzyme activation by ST was unaffected by various extraction procedures. The possibility that the ST receptor and particulate guanylate cyclase were the same molecule was explored. ST binding and cyclic GMP production were separated by affinity chromatography on GTP-agarose. Similarly, guanylate cyclase migrated as a 300,000-dalton protein, while the ST receptor migrated as a 240,000-dalton protein on gel filtration chromatography. Also, thiol-reactive agents such as cystamine and N-ethylmaleimide inhibited guanylate cyclase activation by ST, with no effect on receptor binding of ST. These data suggest that guanylate cyclase and the ST receptor are independent proteins coupled by cytoskeletal components in membranes of intestinal mucosa.     

Influence of age on the production and regulation of interleukin-1 in mice.

The decrease in T-cell proliferation with age is due, in part, to the decline in the production of IL-2. Since IL-1 is needed to trigger IL-2 production, we determined the IL-1 producing capacity of peritoneal macrophages of young (2-4 months) and old (24-26 months) BALB/c and C57BL/6 mice. Mice were stimulated with LPS, and their peritoneal macrophages were obtained 3 days later, purified, and assessed for IL-1 production by coculturing them with splenic T cells at a ratio of 1:5 in the presence of LPS. Supernatants were obtained 4 days later when the PGE2 and IL-2 activities were minimal and IL-1 activity maximal. IL-1 activity was assessed for their ability to augment the proliferative activity of indicator thymocytes in their response to PHA stimulation. The results revealed that (i) IL-1 production by cells of old BALB/c and C57BL/6 mice is reduced to about 40% and 30% that of young mice, respectively; (ii) indomethacin enhances IL-1 production by cells of both young and old mice to the same extent; and (iii) reduction in the IL-1 producing capacity by cells of old mice results from altered activities of both the IL-1 producing peritoneal macrophages and the augmenting T cells.     

Myocardial and coronary vascular responses to insulin in the diabetic lamb.

Diabetes mellitus was produced in 9 lambs by giving alloxan monohydrate, 150 mg/kg, 24 h prior to study, and these were compared with 12 control animals. Responses to insulin, 20 U/kg, were studied in each. Measurements of LV dP/dtmax, coronary sinus flow (CF) and myocardial extraction and uptake of O2, glucose and fatty acids (NEFA) were made using a hemodynamically controlled preparation described previously (Am. J. Physiol. 244: 1381, 1973). Initial arterial glucose (G) averaged 328 mg/dl in the diabetics compared with 110 mg/dl in controls. G fell to 160 mg/dl 90 min after insulin in the diabetics, and to 25 mg/dl in the controls. Initial LV dP/dtmax values were identical in both groups and showed similar increases of about 800 mmHg/s 30 min after insulin (P less than 0.001). However, initial CF was lower and resistance higher in diabetics than controls. After insulin, CF increased 50% in controls but only 10% in diabetics. Because inotropic responsiveness and O2 metabolism were the same in both groups, altered coronary vascular smooth muscle reactivity in the diabetics may explain these findings.     

Observer variability in the histopathological reporting of abnormal rectal biopsy specimens.

AIMS--To study the consistency of reporting of abnormal rectal biopsy specimens, especially in the differentiation of inflammatory bowel disease from other causes of abnormality. METHODS--Sixty rectal biopsy specimens were identified from patients presenting with bloody diarrhoea. These were then circulated to the 11 consultant pathologists in the study who filled in a proforma with a list of 12 diagnostic categories and 22 features. RESULTS--Forty one of the 60 cases were examples of inflammatory bowel disease. In 33 of these cases nine or more pathologists had made the diagnosis. Further categorisation into ulcerative colitis and Crohn''s disease showed better recognition of ulcerative colitis. In the 19 cases of non-inflammatory bowel disease recognition of pseudomembranous colitis and solitary rectal ulcer syndrome was good, but the results were poorer in the case of infective colitis. CONCLUSION--The findings suggest that a group of consultant pathologists can differentiate between inflammatory bowel disease and other causes of an abnormal rectal biopsy specimen and can also recognise pseudomembranous colitis and solitary rectal ulcer syndrome satisfactorily.     

Effects of Purified Staphylococcal Alpha Toxin on the Ultrastructure of Human and Rabbit Erythrocytes

Previous studies have suggested that the primary site of action of purified staphylococcal alpha toxin is the cell membrane. Scanning and transmission electron microscopy studies were undertaken, therefore, to define toxin-induced alterations in the surface morphology of rabbit and human red blood cells. During the prelytic lag phase, scanning electron microscopy revealed multiple discrete blisters on the surface of rabbit red blood cells; during hemolysis, cellular collapse and ghosts were seen, but most striking was the separation of large fragments of cell membrane from red blood cell surfaces. In contrast, alterations in less sensitive human red blood cells were limited to occasional fingerlike protrusions during the period of accelerated lysis. Transmission electron microscopy substantiated these changes. These studies have provided further evidence that the cell membrane is the primary site of action of staphylococcal alpha toxin.     

Controlled elimination of intracellular H(2)O(2): regulation of peroxiredoxin, catalase, and glutathione peroxidase via post-translational modification

The predominant enzymes responsible for elimination of hydrogen peroxide (H(2)O(2)) in cells are peroxiredoxins (Prxs), catalase, and glutathione peroxidases (GPxs). Evidence suggests that catalytic activities of certain isoforms of these H(2)O(2)-eliminating enzymes are extensively regulated via posttranslational modification. Prx I and Prx II become inactivated when phosphorylated on Thr(90) by cyclin B-dependent kinase Cdc2. In addition, the active-site cysteine of Prx I-IV undergoes a reversible sulfinylation (oxidation to cysteine sulfinic acid) in cells. Desulfinylation (reduction to cysteine) is achieved by a novel enzyme named sulfiredoxin. c-Abl and Arg nonreceptor protein tyrosine kinases associate with catalase in cells treated with H(2)O(2) by mechanisms involving the SH3 domains of the kinases and the Pro(293)PheAsnPro motif of catalase and activate catalase by phosphorylating it on Tyr(231) and Tyr(386). Similarily, GPx1 is activated by c-Abl- and Arg-mediated phosphorylation. The tyrosine phosphorylation is critical for ubiquitination-dependent degradation of catalase.     

Weighted expectation maximization reconstruction algorithms with application to gated megavoltage tomography

We propose and investigate weighted expectation maximization (EM) algorithms for image reconstruction in x-ray tomography. The development of the algorithms is motivated by the respiratory-gated megavoltage tomography problem, in which the acquired asymmetric cone-beam projections are limited in number and unevenly sampled over view angle. In these cases, images reconstructed by use of the conventional EM algorithm can contain ring- and streak-like artefacts that are attributable to a combination of data inconsistencies and truncation of the projection data. By use of computer-simulated and clinical gated fan-beam megavoltage projection data, we demonstrate that the proposed weighted EM algorithms effectively mitigate such image artefacts.     

Xanthii Fructus Inhibits Inflammatory Responses in LPS-Stimulated Mouse Peritoneal Macrophages

Xanthii Fructus (XF) is an herb widely used in medicine for the treatment of a variety of inflammatory pathologies. In this study, using mouse peritoneal macrophages, we have examined whether XF affects nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin (IL)-12p40 production induced by interferon (IFN)-γ and lipopolysaccharide (LPS). XF inhibits IFN-γ and LPS-induced NO production in a dose dependent manner. The decrease in NO synthesis was reflected as a decreased amount of inducible NO synthase protein. Furthermore, we also found that XF inhibits pro-inflammatory cytokine TNF-α production. However, treatment of XF in peritoneal macrophages had no effect on IL-12p40 production. These findings suggest that XF may be used in controlling macrophages-mediated inflammatory diseases.