Development of in silico models to guide the experimental characterisation of penile tissue and inform surgical treatment of erectile dysfunction.

This paper presents a computational study to investigate the mechanical properties of human penile tissues. Different experimental testing regimes, namely indentation and plate-compression tests, are compared to establish the most suitable testing regime for establishing the mechanical properties of the different penile tissues. An idealised MRI-based geometry of the penis, containing different tissue layers, is simulated using the finite element (FE) method to enable realistic predictions of the deformation of the penis. Unlike the linear elastic models used in the literature to-date, hyperelastic isotropic/anisotropic material models are used to capture material nonlinearity and anisotropy. The influence of material properties, morphological variations, material nonlinearity and anisotropy are investigated. Moreover, the implantation of an inflatable penile prosthesis (IPP) is simulated to assess the effects of the implantation procedure, material nonlinearity, and anisotropy on tissue stresses. The results indicate that the interior layers of the penis do not affect the overall stiffness of the penis in the indentation test, while the plate-compression test is able to capture the effects of these layers. Tunica Albuginea (TA) is found to have the most significant contribution to the total stiffness of the penis under load. It can also be observed that buckling occurs in the septum of the penis during the compression tests, and different morphologies dictate different compressive behaviours. There is a clear need for future experimental studies on penile tissues given the lack of relevant test data in the literature. Based on this study, plate-compression testing would offer the most insightful experimental data for such tissue characterisation.

Self-reported vividness of tactile imagery for object properties and body regions: An exploratory study.

Mental imagery ability has been examined principally in the visual domain. Despite evidence for tactile mental representations in the absence of direct stimulation, this ability is poorly understood. We investigated tactile imagery for both active and passive tasks in a large sample (N = 118). Vividness of imagery was tested across two different tasks: somatosensory imagery (of body sensitivity) and tactile imagery (of object properties) in all participants. Evidence for vivid imagery across tactile and somatosensory dimensions was found with a positive, albeit weak, correlation in imagery strength between dimensions. Imagery ratings varied across objects and object properties in the tactile imagery task and across body sites in the somatosensory imagery task. These findings shed light on the capacity for, and characteristics of, tactile mental imagery in the general population and suggest that the ability to experience vivid tactile mental images may mediate performance across a number of perceptual tasks.

Pointing perception is precise.

The spontaneity and ease with which we point understates the gesture's significance to understanding cognition. Onset of pointing in infancy predicts early word acquisition and signals a capacity for shared intentionality. Yet, notwithstanding its importance, there is little research on the perception of pointing and its referents. Here we show that perceptual acuity for discerning where another person is pointing is remarkably accurate. Thresholds, as low as 0.5° of visual angle across an interpersonal distance of ∼2 m, are modulated by the referent's location in space and the hand used to point and remain constant when the pointer's eyes are occluded from view and when 'embodiment' cues are enhanced or minimized. Pointing with the index finger not only directs attention toward a general region of space but the morphology of arm, hand and finger can be used to discern the location of the pointer's attention with precision.

Characterization of the expression and inflammatory activity of NADPH oxidase after spinal cord injury.

Reactive oxygen species (ROS) and the NADPH oxidase (NOX) enzyme are both up-regulated after spinal cord injury (SCI) and play significant roles in promoting post-injury inflammation. However, the cellular and temporal expression profile of NOX isotypes, including NOX2, 3, and 4, after SCI is currently unclear. The purpose of this study was to resolve this expression profile and examine the effect of inhibition of NOX on inflammation after SCI. Briefly, adult male rats were subjected to moderate contusion SCI. Double immunofluorescence for NOX isotypes and CNS cellular types was performed at 24 h, 7 days, and 28 days post-injury. NOX isotypes were found to be expressed in neurons, astrocytes, and microglia, and this expression was dependent on injury status. NOX2 and 4 were found in all cell types assessed, while NOX3 was positively identified in neurons only. NOX2 was the most responsive to injury, increasing in both microglia and astrocytes. The biggest increases in expression were observed at 7 days post-injury and increased expression was maintained through 28 days. NOX2 inhibition by systemic administration of gp91ds-tat at 15 min, 6 h or 7 days after injury reduced both pro-inflammatory cytokine expression and evidence of oxidative stress in the injured spinal cord. This study therefore illustrates the regional and temporal influence on NOX isotype expression and the importance of NOX activation in SCI. This information will be useful in future studies of understanding ROS production after injury and therapeutic potentials.

Improving venous thromboembolic disease prophylaxis in medical inpatients: a role for education and audit.

BACKGROUND: Venous thromboembolic disease (VTED) prophylaxis is a key strategy in reducing preventable deaths in medical inpatients. We assessed compliance with internationally published guidelines for VTED prophylaxis in at-risk medical patients before and 1 month after an educational intervention to enhance compliance with such guidelines. RESULTS: One hundred and fifty patients were assessed on each occasion. Pre-intervention, VTED prophylaxis was prescribed in only 48% of at-risk cases. Compliance was best among patients under stroke services and worst for those under acute medical teams. Patients within specialist units were more likely to be prescribed prophylaxis than those in general wards (75 vs. 53%; p = 0.0019). Post-intervention, overall compliance improved to 63% (p = 0.041 for comparison). There was a significant improvement among general medical teams (48 vs. 75%; p = 0.001), and in general wards (52 vs. 74%; p = 0.003). CONCLUSIONS: Thromboprophylaxis is under-prescribed in medical inpatients, but compliance with international guidelines can be significantly enhanced with targeted educational intervention.

Urinary albumin and insulin as predictors of coronary artery disease: An angiographic study.

Microalbuminuria has been associated with cardiovascular risk factors, events, and mortality. It also clusters with hyperinsulinemia and the metabolic syndrome. How urinary albumin excretion and the fasting serum insulin level relate to coronary artery disease (CAD) has not been previously determined. In 308 patients undergoing elective coronary angiography, the albumin to creatinine ratio was measured in urine from an early morning void. The fasting serum insulin level was also determined. CAD was assessed by angiography. Urinary albumin excretion was 28 +/- 5 mg/g (mean +/- SE) in patients with CAD and 10 +/- 1 mg/g in those without CAD (P < 0.001). Fasting serum insulin levels were also greater in patients with CAD compared with those without CAD; 20 +/- 3 and 13 +/- 1 microU/mL, respectively (P = 0.016). Urinary albumin excretion and fasting serum insulin levels increased progressively with severity of CAD. In patients without diabetes (n = 255), significant relationships of urinary albumin excretion and the fasting serum insulin levels to CAD were observed, but they were more pronounced when patients with diabetes (n = 53) were included. In multiple regression analysis, the odds ratios for severe CAD were 2.2 (95% confidence interval, 1.1 to 4.5) for microalbuminuria and 2. 2 (95% confidence interval, 1.3 to 3.8) for hyperinsulinemia. In summary, urinary albumin excretion and the fasting serum insulin levels were directly related to angiographic evidence of CAD. Microalbuminuria and hyperinsulinemia predict a significantly elevated risk for coronary atherosclerosis.

Isolation of the GA-response mutant sly1 as a suppressor of ABI1-1 in Arabidopsis thaliana.

Seed dormancy and germination in higher plants are partially controlled by the plant hormones abscisic acid (ABA) and gibberellic acid (GA). ABA establishes dormancy during embryo maturation, whereas GA breaks dormancy and induces germination. Previous attempts to identify GA response genes were confounded because GA mutants are not expected to germinate and, unlike GA auxotrophs, should fail to be rescued by exogenous GA. Here, we describe a screen for suppressors of the ABA-insensitive mutant ABI1-1 that enriches for GA auxotrophs and GA-insensitive mutants. The vast majority (76%) of the suppressors of ABI1-1 strongly resemble GA auxotrophs in that they are severely dwarfed and have dark green foliage and flowers with underdeveloped petals and stamen. Three isolates were alleles of the GA auxotroph ga1. The remaining severe dwarves were not rescued by GA and belong to a single complementation group that we designate sly1 (Sleepy 1). The alleles of sly1 identified are the first recessive GA-insensitive mutations to reflect the full spectrum of GA-associated phenotypes, including the failure to germinate in the absence of the ABI1-1 lesion. Thus, we postulate that SLY1 is a key factor in GA reception.

A protein farnesyl transferase involved in abscisic acid signal transduction in Arabidopsis.

The hormone abscisic acid (ABA) modulates a variety of developmental processes and responses to environmental stress in higher plants. A collection of mutations, designated era, in Arabidopsis thaliana that confer an enhanced response to exogenous ABA includes mutations in the Era1 gene, which encodes the beta subunit of a protein farnesyl transferase. In yeast and mammalian systems, farnesyl transferases modify several signal transduction proteins for membrane localization. The era1 mutants suggest that a negative regulator of ABA sensitivity must be acted on by a farnesyl transferase to function.

Insulin secretion and DNA synthesis of cultured islets of Langerhans are influenced by the matrix.

To compare the effect of matrix on glucose-stimulated insulin release, we cultured neonatal (3- to 5-day-old) rat islets of Langerhans, devoid of mesenchymal cell, on fibronectin, Cell-Tak, or endothelial basement membranes, free-floating, or dispersed into single cells. We also examined the rate of DNA synthesis during the culture period. Compared to free-floating islets [0.386 +/- 0.03 (SEM) ng per 24 h/ng total], single-cell cultures had the lowest basal insulin release (0.159 +/- 0.03 ng per 24 h/ng total; p < 0.0001), which was also low in islets attached to endothelial basement membrane (0.294 +/- 0.02 ng per 24 h/ng total; p = 0.01). The spontaneous insulin release (1 h in medium with 2.7 mM glucose) was lowest in islets attached to endothelial basement membrane (0.003 +/- 0.00023 ng per h/ng total; p < 0.0001 vs. free-floating) and highest in single-cell cultures (0.01153 +/- 0.00259 ng per h/ng total; p = 0.039 vs. free-floating). The ability to increase insulin release following a glucose challenge (16.1 mM for 1 h) was highest in islets grown on endothelial basement membranes (16.4-fold) and fibronectin (12.6-fold) compared to free-floating islets (8.7-fold), Cell-Tak (7.9-fold), and single-cell cultures (5.4-fold).(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro growth of neonatal rat islet cells is stimulated by adhesion to matrix.

We examined DNA synthesis in non-enzymatically isolated neonatal rat pancreatic islets sub-cultured to eliminate fibroblast contamination, which was excluded both by demonstrating no effect of fibroblast growth factor (FGF) on 3H-thymidine incorporation and by immunofluorescence of attached islets using a mouse anti-fibroblast monoclonal antibody. 3H-thymidine incorporation in islets increased with increasing glucose up to a concentration of 21.1 mM in both free-floating islets (11,789 cpm/micrograms DNA +/- 1,610 SEM) and islets attached to fibronectin coated plastic (43,043 cpm/micrograms DNA +/- 9,203 SEM). These values were significantly higher when compared to 3H-thymidine incorporation in medium containing 11.1 mM glucose (p < 0.007, and p < 0.0001 for free-floating and attached islets respectively). 3H-thymidine incorporation was significantly higher in attached islets than in free-floating islets at all glucose concentrations tested (p < 0.005 at 11.1 mM, 16.1 mM, and 21.1 mM; and p < 0.01 at 26.1 mM). 5-bromo-2'-deoxyuridine (BrDU) staining of islets showed an increased number of positive nuclei in cells localised within attached islets (37.6 nuclei per islet +/- 5.1 SEM) compared to free-floating islets (7.62 nuclei per islet +/- 1.04 SEM, p < 0.001), indicating that attachment influenced proliferation of islet cells not physically in contact with the matrix. No difference in glucose-stimulated insulin release was observed between attached and free-floating islets. In conclusion, a fibroblast free islet culture was used to document the stimulatory effect of islet attachment on DNA synthesis, which was greater than the stimulation exerted by glucose alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Measurement of PAF in blood by radioimmunoassay. Examination of interfering factors and problems involved in accurate quantification.

A recently developed radioimmunoassay for PAF was applied to blood extracts with the aims of defining and overcoming problems that lead to erroneous results and establishing optimum conditions for the accurate determination of PAF levels. The high lipid content of blood was found to interfere with the assay, and it appeared that phosphatidylcholine and/or sphingomyelin might be among the lipids responsible. Interference was eliminated by either dilution or preparative TLC of blood extract prior to RIA although dilution is unlikely to be generally useful due to the low amounts of PAF normally present in human blood. Lipid extraction of whole blood followed by preparative TLC proved to be necessary in the preparation of samples prior to performance of the RIA. The problems encountered in the measurement of PAF levels in blood by RIA highlight the importance of determining the correct method of sample preparation for any tissue/fluid prior to its inclusion in the assay.

Quantitation by radioimmunoassay of PAF in human saliva.

Approximately 75% of the PAF present in saliva is recovered on extraction of whole saliva (0.8 vol) with chloroform/methanol/water (2:2:1, v/v/v). PAF levels, determined by our recently developed radioimmunoassay, in saliva extracts ranged from 0.5-21 ng/mL with 59% between 2-6 ng/mL. These figures, for apparently healthy subjects, are higher than previously reported levels obtained by platelet assays. The validity of our radioimmunoassay results was checked by isolating and quantitating the PAF fraction from whole saliva. In addition, when we examined our saliva samples by platelet aggregation, low levels of PAF, comparable with the values found in the literature, were detected. Investigations revealed the presence of a substance(s) which inhibited PAF-induced platelet aggregation but which did not affect the radioimmunoassay.

Antibodies to platelet-activating factor (PAF) inhibit PAF-induced platelet aggregation.

Platelet-activating factor (PAF) is the most potent platelet agonist known. PAF-induced platelet aggregation was blocked by pre-incubation of PAF with the immunoglobulin fraction from sheep or rabbit anti-PAF anti-serum. Inhibition was specific for PAF and was dependent upon immunoglobulin and PAF concentrations. Antibody-mediated inhibition of PAF-activity may be a valuable method for studying the biological effects of PAF in some systems.

Stability of platelet activating factor (PAF) in human saliva. Quantitation by radioimmunoassay.

Platelet activating factor (PAF) is thought to mediate many inflammatory processes and its involvement in health and disease may be clarified by examining PAF levels in human secretions. The known presence of PAF, the ease of obtaining samples and the relative stability of PAF in saliva, makes this fluid a preferred source for examination of PAF in health and disease. The activity of PAF-acetylhydrolase (the PAF degrading enzyme) in saliva was 1,000-fold lower than that found in human plasma. Extraction of saliva with chloroform/methanol/water resulted in 70-90% recovery of PAF. Using the radioimmunoassay (RIA), PAF levels in the range 0.5-21 ng/ml were found in normal human salivas. These values were significantly higher than those reported from bioassay studies based on washed platelets. The validity of the RIA was checked by isolating and quantitating the PAF fraction from whole saliva extract, and by treatment of the extracts with the enzyme phospholipase A2. Direct comparison of salivary PAF levels, determined by both platelet aggregation (PA) and RIA confirmed our original finding that values obtained were lower using the bioassay method. Furthermore, these bioassay values compared favourably with those in the literature. Investigations revealed the presence of a substance(s) in saliva which inhibited PAF-induced platelet aggregation but which did not affect the radioimmunoassay.

Examination for platelet-activating factor production by preimplantation mouse embryos using a specific radioimmunoassay.

A specific and highly sensitive radioimmunoassay was used to measure platelet-activating factor (PAF) production by preimplantation mouse embryos in vitro. Levels of PAF greater than 1 pg per embryo were not observed in 24-h culture medium from 2-cell embryos, compacted morulae or blastocysts, or in extracts from these embryos. Synthetic PAF added to embryos at the start of culture could be almost totally recovered after the incubation period, indicating negligible degradation of PAF during culture. PAF was also not detected in embryo samples using a washed rabbit-platelet aggregation assay. It can be concluded that mouse embryos do not produce substantial levels of PAF, or any of the biologically active analogues of PAF detected by the assay.

Combining site specificities of rabbit antibodies to platelet-activating factor (PAF).

Anti-PAF sera from six different rabbits, immunized with C12- or C6-PAF as immunogen, were examined in hapten inhibition experiments in an attempt to define the fine structural recognition specificities of the antibody combining sites. Using a selection of naturally occurring lipids and PAF analogues, no significant cross-reactivity was observed with the lipids or with the inactive metabolite, lyso-PAF. Comparison of the structural specificity requirements of the antibodies from each rabbit showed some heterogeneity, with one antiserum demonstrating a different recognition specificity at position 1 on the glycerol backbone of the PAF molecule. A second rabbit antiserum showed a large degree of tolerance for analogues with increasing acyl chain length at position 2. In general, an ether group at position 1 and an acetyl at position 2 were required for inhibitory activity and a degree of tolerance was demonstrated at position 3, where the main structural requirement was for one or more methyl groups on the nitrogen atom of the phosphocholine moiety.