A test of the Energetics-Hormone Vocalization model in the green treefrog.

Male courtship displays may be regulated by, and affect the production of, circulating hormones. The Energetics-Hormone Vocalization (EHV) model, for example, posits that interactions among chorusing male anuran amphibians stimulate androgen production that then mediates an increase in vocal effort. Increased vocal effort is expected to deplete energy reserves and increase glucocorticoid levels that, in turn, negatively affect androgen levels and vocalization. Androgen levels, glucocorticoid levels, and vocal effort are thus expected to increase across and within nights of chorus activity and should be positively correlated in calling males; energy reserves should decline temporally and be inversely related to glucocorticoid levels. We tested predictions of the EHV model in the green treefrog, Hyla cinerea. Consistent with the model, both testosterone and dihydrotestosterone levels increased across the breeding season in calling males. However, testosterone levels decreased and dihydrotestosterone levels did not change within nights of chorus activity, suggesting that chorusing behavior did not drive the seasonal elevation in androgens. Corticosterone (CORT) level remained relatively stable across the breeding season and decreased within nights of chorus activity, contrary to model predictions. Body condition, the proxy for energetic state, was inversely correlated with CORT level but discrepancies between model predictions and temporal patterns of CORT production arose because there was no evidence of a temporal decrease in body condition or increase in vocal effort. Moreover, androgen and CORT levels were not positively correlated with vocal effort. Additional ecological and physiological measures may be needed to support predictions of the EHV model.

Regression of lung cancer by hypoxia-sensitizing ruthenium polypyridyl complexes.

The ruthenium (II) polypyridyl complexes (RPC), Δ-[(phen)2Ru(tatpp)]Cl2 (Δ-[3]Cl2) and ΔΔ-[(phen)2Ru(tatpp)Ru(phen)2]Cl4 (ΔΔ-[4]Cl4, are a new generation of metal-based antitumor agents. These RPCs bind DNA via intercalation of the tatpp ligand, which itself is redox-active and is easily reduced at biologically relevant potentials. We have previously shown that RPC 4(4+) cleaves DNA when reduced by glutathione to a radical species and that this DNA cleavage is potentiated under hypoxic conditions in vitro. Here, we show that 3(2+) also exhibits free radical-mediated DNA cleavage in vitro and that 3(2+) and 4(4+) both exhibit selective cytotoxicity toward cultured malignant cell lines and marked inhibition of tumor growth in vivo. The murine acute toxicity of RPCs 3(2+) and 4(4+) (maximum tolerable doses ~ 65 µmol/kg) is comparable with that for cisplatin (LD50 ~ 57 µmol/kg), but unlike cisplatin, RPCs are generally cleared from the body unchanged via renal excretion without appreciable metabolism or nephrotoxic side effects. RPCs 3(2+) and 4(4+) are shown to suppress growth of human non-small cell lung carcinoma (~83%), show potentiated cytotoxicity in vitro under hypoxic conditions, and induce apoptosis through both intrinsic and extrinsic pathways. The novel hypoxia-enhanced DNA cleavage activity and biologic activity suggest a promising new anticancer pharmacophore based on metal complexes with aromatic ligands that are easily reduced at biologically accessible potentials.

Characterization of renal ischemia using DLP hyperspectral imaging: a pilot study comparing artery-only occlusion versus artery and vein occlusion.

BACKGROUND AND PURPOSE: Renal artery-only (AO) occlusion, as opposed to artery and vein (AV) occlusion, has demonstrated some benefit in reducing renal insufficiency during warm ischemia. In this pilot study, we used digital light projection hyperspectral imaging (HSI) to construct a "real time" tissue oxygenation "map" to determine whether there are differences in renal tissue oxygenation during vascular occlusion with AO vs AV. MATERIALS AND METHODS: Renal vascular occlusion with either AO or AV was performed for 60 minutes in seven porcine renal units. Using HSI, the percentage of oxyhemoglobin (%HbO(2)) in the renal cortex was determined at 4-minute increments throughout the ischemic period and for 30 minutes after reperfusion. RESULTS: Average baseline %HbO(2) in all animals was approximately 70%. After vascular occlusion in both cohorts, %HbO(2) decreased by one third within 2 to 5 minutes, with a gradual decline in %HbO(2) over the remaining 55 minutes. Oxyhemoglobin profiles for AO and AV occlusion diverged significantly between 16 and 24 minutes after vascular occlusion (P = 0.0001 and 0.036, respectively), with a merging of the two curves occurring after approximately 36 minutes (P = 0.093). During reperfusion, average %HbO(2) improved to 72.4% after 25 to 30 minutes. CONCLUSION: In this pilot study, we demonstrate that renal tissue oxygenation drops rapidly after occlusion of the renal vasculature and returns to near baseline 30 minutes after reperfusion. In the porcine model, the %HbO(2) differs significantly between AO and AV occlusion for up to 35 minutes after ischemia onset, indicating a possible "ischemic window" in which AO occlusion may provide benefit over AV occlusion.

Intraoperative bile duct visualization using near-infrared hyperspectral video imaging.

BACKGROUND: Current methodologies for imaging the biliary system during cholecystectomy are cumbersome and do not eliminate the risk of bile duct injury. We describe an approach to intraoperative biliary imaging that will enable surgeons to see through the hepatoduodenal ligament and visualize the anteriorly placed biliary system. METHODS: A laparoscopic-capable, near-infrared, hyperspectral imaging system was built. Reflected light passes through a liquid crystal filter that is continuously tunable in the near-infrared spectrum (650-1,100 nm). Spectroscopic image data are collected from laparoscopic surgery images onto array detectors formatted into a 3-dimensional hyperspectral data cube having spatially resolved images in the x-y plane and wavelength data in the z plane. Deconvoluting and color-coding the spatial and spectral information provides an image representative of inherent chemical properties to the imaged tissue. RESULTS: Images of porcine biliary structures were obtained. The common duct-reflected spectra displayed a characteristic lipid shoulder at 930 nm and a strong water peak at 970 nm. Venous structures had absorption peaks at 760 nm (deoxyhemoglobin), 800 nm (oxyhemoglobin), and 970 nm (water). Arterial vessels had absorption peaks at 800 nm and 970 nm that would be expected for oxyhemoglobin and water. CONCLUSIONS: We have designed and constructed a device to significantly enhance intraoperative biliary imaging. This system should enable surgeons to see through the hepatoduodenal ligament and image the anteriorly placed biliary system without the need for dissection of the cystic duct, as is needed with intraoperative cholangiography. Because the biliary system can be seen before any dissection is performed, this dimensional imaging technology has the potential for eradicating bile duct injury.

Exploration of structure-antifouling relationships of capsaicin-like compounds that inhibit zebra mussel (Dreissena polymorpha) macrofouling.

Macrofouling of aquatic man-made structures by zebra mussels (Dreissena polymorpha) poses significant economic burdens on commercial freshwater shipping and facilities utilising raw water. The negative environmental impact of some current antifouling technologies has limited their use and prompted investigation of non-organometallic and non-oxidising antifoulants as possible environment-friendly alternatives. The plant-derived natural product capsaicin and 18 other compounds with one or more capsaicin-like structural features were tested for their potential to inhibit zebra mussel byssal attachment at a single high concentration of 30 microM. Of these, three compounds displaying the highest levels of attachment inhibition where selected for further concentration-response testing. This testing revealed that capsaicin (8-methyl-N-vanillyl-trans-6-nonenamide), N-vanillylnonanamide, and N-benzoylmonoethanolamine benzoate all inhibited byssal attachment with potency values (EC(50)) in the micromolar range. None of these compounds were lethal to adult specimens of the water flea, Daphnia magna, at concentrations that inhibited mussel byssal attachment.

Characterization of a near-infrared laparoscopic hyperspectral imaging system for minimally invasive surgery.

We developed and characterized a new imaging platform for minimally invasive surgical venues, specifically a system to help guide laparoscopic surgeons to visualize biliary anatomy. This platform is a novel combination of a near-infrared hyperspectral imaging system coupled with a conventional surgical laparoscope. Intraoperative tissues are illuminated by optical fibers arranged in a ring around a center-mounted relay lens collecting back-reflected light from tissues to the hyperspectral imaging system. The system consists of a focal plane array (FPA) and a liquid crystal tunable filter, which is continuously tunable in the near-infrared spectral range of 650-1100 nm with the capability of passing light with a mean bandwidth of 6.95 nm, and the FPA is a high-sensitivity back-illuminated, deep depleted charge-coupled device. Placing a standard resolution target 5.1 cm from the distal end of the laparoscope, a typical intraoperative working distance, produced a 7.6-cm-diameter field of view with an optimal spatial resolution of 0.24 mm. In addition, the system's spatial and spectral resolution and its wavelength tuning accuracy are characterized. The spectroscopic images are formatted into a three-dimensional hyperspectral image cube and processed using principle component analysis. The processed images provide contrast based on measured spectra associated with chemically different anatomical structures helping identify the main molecular chromophores inherent to each tissue. The principal component images were found to image swine gallbladder and biliary structures from surrounding tissues, in real time, during cholecystectomy surgery. Furthermore, it is shown that surgeons can interrogate selected image subregions for their molecular composition identifying biliary anatomy during surgery and before any invasive action is undertaken.

Sex-biased gene expression in a ZW sex determination system.

Studies of the transcriptome have shown that a substantial fraction of interspecific differences in gene expression is the result of sex-biased gene expression. These results suggest that sex-dependent selection may be an important force in generating differences between species but to date all studies have focused on Drosophila. We examined a sample of the transcriptome in the gonads of two species of Xenopus to provide an additional test of how sex-biased gene expression may contribute to differences in gene expression between species. In contrast to Drosophila, Xenopus provides an example of a ZW system with morphologically indistinguishable sex chromosomes. About 81% of the transcriptome was differentially expressed between X. laevis and X. muelleri and there were more genes that were male-biased compared to the number of genes that were female-biased or non-sex-biased. While there were more genes up-regulated in males of Xenopus, the largest magnitude difference in expression between species occurred in female-biased genes, and male-biased genes were proportionally more abundant for the smallest changes in expression between species. Our results suggest that more genes have a role in the function of the testis compared to the ovary and female-biased genes play a principle role in expression divergence between species. These results differ from those in the Drosophila XY system in that more female-biased genes had >4-fold difference of expression between species than male-biased genes, suggesting that ZW sex chromosomes may facilitate enhanced gene expression divergence between species.

Dorsal horn neuron response patterns to graded heat stimuli in the rat.

Sensory input from various receptors in the periphery first becomes integrated in the spinal cord dorsal horn. The response of the spinal cord dorsal horn neurons to mechanical stimuli are classified as low threshold, high threshold, and wide dynamic range neurons. However, the response pattern of deep dorsal horn cells to heat has not been well described. In this study, the response of the spinal cord dorsal horn neurons to graded heat stimuli were characterized in 147 neurons in rats by extracellular single cell recording. After a differentiable cell was identified, the Peltier heat stimulator was applied to the receptive field and the base temperature was set at 30 degrees C. The heat stimulus was delivered for 10 s from 37-51 degrees C in 2 degrees C increments, with an inter-stimulus interval of 30 s. Out of the 147 neurons, five statistically distinguishable response patterns were identified by latent class cluster analysis. It is concluded that variation of temperature may account for the observed results and indicate functionally different subsets of heat-responsive cells in the deep dorsal horn.