Expression of integrin ανß6 differentiates perihilar cholangiocarcinoma (PHC) from benign disease mimicking PHC.

BACKGROUND: Approximately 15% of patients undergoing resection for presumed perihilar cholangiocarcinoma (PHC) have benign disease at final pathological assessment. Molecular imaging targeting tumor-specific biomarkers could serve as a novel diagnostic tool to reduce these futile surgeries. Imaging agents have been developed, selectively binding integrin ανß6, a cell receptor upregulated in pancreatobiliary malignancies, for both (preoperative) PET and (intraoperative) fluorescent imaging. Here, expression of integrin ανß6 is evaluated in PHC, intrahepatic cholangiocarcinoma (ICC), hepatocellular carcinoma (HCC) and benign disease mimicking PHC using immunohistochemistry. MATERIALS & METHODS: Three tissue microarrays (TMA) including 103 PHC tumor cores and sixty tissue samples were selected from resection specimens of pathologically proven PHC (n = 20), ICC (n = 10), HCC (n = 10), metastatic PHC lymph nodes (n = 10) and benign disease (presumed PHC with benign disease at pathological assessment, n = 10). These samples were stained for integrin ανß6 and quantified using the H-score. RESULTS: Immunohistochemical staining for integrin ανß6 showed membranous expression in all twenty PHC whole mount slides (100%) and 93 out of 103 (92%) PHC tumor cores. Mean H-score of PHC samples was 195 ± 71, compared to a mean H-score of 126 ± 57 in benign samples (p = 0.013). In both benign and PHC samples, inflammatory infiltrates and pre-existent peribiliary glands showed integrin ανß6 expression. The mean H-score across ten ICC was 33 ± 53, which was significantly lower compared to PHC (p < 0.001) but too weak to consistently discriminate ICC from HCC (H-score 0)(p = 0.062). CONCLUSION: Integrin ανß6 is abundantly expressed in PHC and associated metastatic lymph nodes. Expression is significantly higher in PHC as compared to benign disease mimicking PHC, ICC and HCC, emphasizing its potential as a target for tumor-specific molecular imaging.

An innovative technique for electronic transport model of group-III nitrides.

An optimized empirical pseudopotential method (EPM) in conjunction with virtual crystal approximation (VCA) and the compositional disorder effect is used for simulation to extract the electronic material parameters of wurtzite nitride alloys to ensure excellent agreement with the experiments. The proposed direct bandgap results of group-III nitride alloys are also compared with the different density functional theories (DFT) based theoretical results. The model developed in current work, significantly improves the accuracy of calculated band gaps as compared to the ab-initio method based results. The physics of carrier transport in binary and ternary nitride materials is investigated with the help of in-house developed Monte Carlo algorithms for solution of Boltzmann transport equation (BTE) including nonlinear scattering mechanisms. Carrier-carrier scattering mechanisms defined through Coulomb-, piezoelectric-, ionized impurity-, surface roughness-scattering with acoustic and intervalley scatterings, all have been given due consideration in present model. The direct and indirect energy bandgap results have been calibrated with the experimental data and use of symmetric and asymmetric form factors associated with respective materials. The electron mobility results of each binary nitride material have been compared and contrasted with experimental results under appropriate conditions and good agreement has been found between simulated and experimental results.

Effect of Mirtazapine on Gastric Emptying in Patients with Cancer-associated Anorexia.

BACKGROUND/AIMS: The tetracyclic antidepressant mirtazapine is widely used in cancer patients suffering from anorexia. Although it is known to restore appetite, the exact mechanism remains unknown. The aim of the study was to evaluate if mirtazapine has any effect on gastric emptying in patients suffering from cancer-related anorexia. MATERIALS AND METHODS: Solid-meal gastric-emptying study using radiolabeled meal was performed in 28 patients suffering from cancer anorexia once at baseline and repeated after 15 days of mirtazapine therapy. RESULTS: At baseline, only 7 (25%) patients had normal gastric motility (emptying >70% at 3 h postingestion) whereas after treatment, 18 (64.2%) patients achieved this limit. Mean % gastric emptying increased from 55.2% ±21.0% to 68.9% ±21.3% (P < 0.001). Mean gastric emptying time (t1/2) before intervention was 314.7 ± 421.0 min which decreased to 116.0 ± 106.7 min after intervention. Results were further analyzed by dividing the patients into two groups based on baseline gastric-emptying study. Group A (normal gastric emptying) consisted of seven patients, mean % gastric emptying at baseline and postintervention was 75.0% ±5.25% and 87.57% ±5.94%, respectively (P < 0.018). Group B (delayed gastric emptying) consisted of 21 patients, mean % gastric emptying at baseline and postintervention was 48.71% ±18.82% and 62.76% ±16.86%, respectively (P < 0.001). CONCLUSION: Mirtazapine significantly improves gastric emptying in patients of prostate and breast cancer suffering from cancer-associated anorexia.

Processable conducting graphene/chitosan hydrogels for tissue engineering.

Composites of graphene in a chitosan-lactic acid matrix were prepared to create conductive hydrogels that are processable, exhibit tunable swelling properties and show excellent biocompatibility. The addition of graphene to the polymer matrix also resulted in significant improvements to the mechanical strength of the hydrogels, with the addition of just 3 wt% graphene resulting in tensile strengths increasing by over 200%. The composites could be easily processed into three-dimensional scaffolds with finely controlled dimensions using additive fabrication techniques and fibroblast cells demonstrate good adhesion and growth on their surfaces. These chitosan-graphene composites show great promise for use as conducting substrates for the growth of electro-responsive cells in tissue engineering.

Syntheses and discovery of a novel class of cinnamic hydroxamates as histone deacetylase inhibitors by multimodality molecular imaging in living subjects.

Histone deacetylases (HDAC) that regulate gene expression are being explored as cancer therapeutic targets. In this study, we focused on HDAC6 based on its ability to inhibit cancerous Hsp90 chaperone activities by disrupting Hsp90/p23 interactions. To identify novel HDAC6 inhibitors, we used a dual-luciferase reporter system in cell culture and living mice by bioluminescence imaging (BLI). On the basis of existing knowledge, a library of hydrazone compounds was generated for screening by coupling cinnamic hydroxamates with aldehydes and ketones. Potency and selectivity were determined by in vitro HDAC profiling assays, with further evaluation to inhibit Hsp90(α/ß)/p23 interactions by BLI. In this manner, we identified compound 1A12 as a dose-dependent inhibitor of Hsp90(α/ß)/p23 interactions, UKE-1 myeloid cell proliferation, p21(waf1) upregulation, and acetylated histone H3 levels. 1A12 was efficacious in tumor xenografts expressing Hsp90(α)/p23 reporters relative to carrier control-treated mice as determined by BLI. Small animal (18)F-FDG PET/CT imaging on the same cohort showed that 1A12 also inhibited glucose metabolism relative to control subjects. Ex vivo analyses of tumor lysates showed that 1A12 administration upregulated acetylated-H3 by approximately 3.5-fold. Taken together, our results describe the discovery and initial preclinical validation of a novel selective HDAC inhibitor.

Light in and sound out: emerging translational strategies for photoacoustic imaging.

Photoacoustic imaging (PAI) has the potential for real-time molecular imaging at high resolution and deep inside the tissue, using nonionizing radiation and not necessarily depending on exogenous imaging agents, making this technique very promising for a range of clinical applications. The fact that PAI systems can be made portable and compatible with existing imaging technologies favors clinical translation even more. The breadth of clinical applications in which photoacoustics could play a valuable role include: noninvasive imaging of the breast, sentinel lymph nodes, skin, thyroid, eye, prostate (transrectal), and ovaries (transvaginal); minimally invasive endoscopic imaging of gastrointestinal tract, bladder, and circulating tumor cells (in vivo flow cytometry); and intraoperative imaging for assessment of tumor margins and (lymph node) metastases. In this review, we describe the basics of PAI and its recent advances in biomedical research, followed by a discussion of strategies for clinical translation of the technique.

MicroRNA-regulated non-viral vectors with improved tumor specificity in an orthotopic rat model of hepatocellular carcinoma.

In hepatocellular carcinoma (HCC), tumor specificity of gene therapy is of utmost importance to preserve liver function. MicroRNAs (miRNAs) are powerful negative regulators of gene expression and many are downregulated in human HCC. We identified seven miRNAs that are also downregulated in tumors in a rat hepatoma model (P<0.05) and attempted to improve tumor specificity by constructing a panel of luciferase-expressing vectors containing binding sites for these miRNAs. Attenuation of luciferase expression by the corresponding miRNAs was confirmed across various cell lines and in mouse liver. We then tested our vectors in tumor-bearing rats and identified two miRNAs, miR-26a and miR-122, that significantly decreased expression in liver compared with the control vector (6.40 and 0.26%, respectively; P<0.05). In tumor, miR-122 had a nonsignificant trend towards decreased (∼50%) expression, whereas miR-26 had no significant effect on tumor expression. To our knowledge, this is the first work using differentially expressed miRNAs to de-target transgene expression in an orthotopic hepatoma model and to identify miR-26a, in addition to miR-122, for de-targeting liver. Considering the heterogeneity of miRNA expression in human HCC, this information will be important in guiding development of more personalized vectors for the treatment of this devastating disease.

Patient-maintained propofol sedation using reaction time monitoring: a volunteer safety study.

Previous volunteer studies of an effect-site controlled, patient-maintained sedation system using propofol have demonstrated a risk of over-sedation. We have incorporated a reaction-time monitor into the handset of the patient-maintained sedation system to add an individualised patient-feedback mechanism. This study assessed if such reaction-time feedback modification would reduce the risk of over-sedation in 20 healthy volunteers deliberately attempting to over-administer themselves propofol. All the volunteers successfully sedated themselves without reaching any unsafe endpoints. All volunteers maintained verbal contact throughout, in accordance with the definition of conscious sedation. The mean (SD) lowest S(p) O(2) was 97 (1.7) % when breathing room air and no volunteer required supplementary oxygen. The mean (SD) maximum effect-site propofol concentration reached was 1.7 (0.4) µg.ml(-1) . The present system was found to be safer than its predecessors, allowing conscious sedation, but preventing over-sedation.

Accidental radiation exposure leading to non-healing ulcers.

Two patients with accidental radiation injury presented at the Sir Ganga Ram Hospital. The first patient, a 41-year-old male, presented with a large necrotic patch on his right gluteal region, which was debrided. Due to the progressive nature of the injury, he developed further necrosis and a non-healing ulcer over the right gluteal region, which was further debrided and covered with a tensor fascia lata flap. Further necrosis over the ischial region was debrided and covered with a pedicled gracilis muscle flap, following which he again required debridement and flap cover for the residual area, which then finally healed. The second patient had a history of radiation exposure followed by a non-healing ulcer on right arm. He was managed by debridement and flap coverage in a single sitting. These two cases suggest that wide excision, thorough debridement and coverage with vascularised tissue are effective in treating radiation ulcers and emphasise that all radiation sources and their management should be strictly controlled.

Comparison of glomerular filtration rate estimated by plasma clearance method with modification of diet in renal disease prediction equation and Gates method.

Glomerular filtration rate (GFR) prediction equations are widely used in clinical practice for quick assessment of kidney function. Gates method using radionuclide technique is an alternative to prediction equations for quick assessment of GFR. Aim of the study was to compare Gates method and modification of diet in renal disease (MDRD) equation in a sizeable patient population with wide range of renal function to evaluate their clinical utility. GFR was estimated in 897 subjects with wide range of renal function by gates method, and MDRD equation and results were compared against measured GFR. Subjects were divided in to 4 groups (0-30 ml, 31-60 ml, 61-90 ml, >90 ml) on the basis of measured GFR and comparison between two methods done through linear regression analysis. Analysis of R(2) indicated that 56% of the interindividual variability for Gates GFR was in accordance to variation in measured GFR, in the GFR range of (0-30 ml), this value dropped to 39% in the GFR range of 31-60 ml, 40% in the GFR range of 61-90 ml, 26.4% in the GFR range of >90 ml, the corresponding figure for MDRD GFR were 47.9%, 31.1%, 17.6% and 16.1%, respectively. Gates method is more precise for GFR estimation at all levels of renal function.

Factors predicting improvement of renal function after pyeloplasty in pediatric patients: a prospective study.

PURPOSE: We assessed the specific preoperative sonographic and urinary factors that may be important in predicting renal function outcomes after pyeloplasty. MATERIALS AND METHODS: We prospectively studied 52 consecutive patients with ureteropelvic junction obstruction who underwent pyeloplasty at our tertiary care center between September 2009 and January 2011. Mean ± 2 SD patient age was 4.26 years (range 3 months to 18 years), and minimum followup was 6 months. Preoperative ultrasound findings recorded were pelvic anteroposterior diameter, pelvic cortical thickness, pelvic volume and pelvic cortical ratio. Spot urine protein-to-creatinine ratio from the renal pelvis and bladder was measured intraoperatively. Based on changes in differential renal function on diuretic renogram, patients were divided into 3 groups. Group 1 had stable differential renal function with less than 5% change, group 2 had improved differential renal function greater than 5% and group 3 had deterioration of differential renal function greater than 5%. Data were analyzed using SPSS®, version 17 with cross-tabulation, nonparametric tests and logistic regression. RESULTS: On ultrasound only anteroposterior diameter (p = 0.018) and pelvic cortical ratio (p = 0.038) were significantly different among the 3 groups. Difference in bladder sample protein-to-creatinine ratio was not significant (p = 0.69), while pelvic urine protein-to-creatinine ratio was significant (p = 0.001). Anteroposterior diameter, pelvic protein-to-creatinine ratio and pelvic cortical ratio were less than 50 mm, 0.5 and 15, respectively, in all patients with improved renal function. CONCLUSIONS: Sonographic and urinary biochemical parameters may predict improvement in renal function after pyeloplasty. Pelvic anteroposterior diameter, pelvic cortical ratio and pelvic urine protein-to-creatinine ratio are the most useful parameters.

A study to improve the image quality in low-dose computed tomography (SPECT) using filtration.

BACKGROUND: The output of the X-ray tube used in computed tomography (CT) provides a spectrum of photon energies. Low-energy photons are preferentially absorbed in tissue; the beam spectrum shifts toward the higher energy end as it passes through more tissue, thereby changing its effective attenuation coefficient and producing a variety of artifacts (beam-hardening effects) in images. Filtering of the beam may be used to remove low-energy photon component. The accuracy of attenuation coefficient calculation by bilinear model depends highly upon accuracy of Hounsfield units. Therefore, we have made an attempt to minimize the beam-hardening effects using additional copper filter in the X-ray beam. The quantitative evaluation were made to see the effect of additional filters on resulting CT images. MATERIALS AND METHODS: This study was performed on dual-head SPECT (HAWKEYE 4, GE Healthcare) with low-dose CT which acquires images at peak voltages of 120/140 kV and a tube current of 2.5 mA. For the evaluation of image quality, we used CT QA Phantom (PHILIPS) having six different density pins of Water, Polyethylene, Nylon (Aculon), Lexan, Acrylic (Perspex) and Teflon. The axial images were acquired using copper filters of various thicknesses ranging from 1 to 5 mm in steps of 1 mm. The copper filter was designed in such a manner that it fits exactly on the collimator cover of CT X-ray tube. Appropriate fixation of the copper filter was ensured before starting the image acquisition. As our intention was only to see the effect of beam hardening on the attenuation map, no SPECT study was performed. First set of images was acquired without putting any filter into the beam. Then, successively, filters of different thicknesses were placed into the beam and calibration of the CT scanner was performed before acquiring the images. The X-ray tube parameters were kept the same as that of unfiltered X-ray beam. All the acquired image sets were displayed using Xeleris 2 (GE Healthcare) on a high-resolution monitor. Moreover, Jaszak's SPECT Phantom after removing the spheres was used to see the different contrast intensities by inserting the different contrast materials of iodine and bismuth in water as background media. Images were analyzed for visibility, spatial resolution and contrast. RESULTS: Successive improvement in the image quality was noticed when we increased the filter thickness from 1 to 3 mm. The images acquired with 3-mm filter appeared almost with no artifacts and were visibly sharper. Lower energy photons from X-ray beam cause a number of artifacts, especially at bone-tissue interfaces. Additional filtrations removed lower energy photons and improved the image quality. Degradation in the image quality was noticed when we increased the filter thickness further to 4 and 5 mm. This degradation in image quality happened due to reduced photon flux of the resulting X-ray beam, causing high statistical noise. The spatial resolution for image matrix of 512 × 512 was found to be 1.29, 1.07, 0.64 and 0.54 mm for without filter, with 1, 2 and 3 mm filters, respectively. The image quality was further analyzed for signal-to-noise ratio (SNR). It was found to be 1.72, 1.78, 1.98 and 1.99 for open, with 1, 2 and 3 mm filters respectively. This shows that 3-mm filter results in an improvement of 15.7% in SNR. CONCLUSION: On the basis of this study, we could conclude that use of 3-mm copper filter in the X-ray beam is optimal for removing the artifacts without causing any significant reduction in the photon flux of the resulting X-ray beam. We also propose that as artifacts have been removed from the images, the value of Hounsfield units will be more accurate and hence the value of attenuation coefficients lead to better contrast and visualization of SPECT images.

Gold nanoparticles: a revival in precious metal administration to patients.

Gold has been used as a therapeutic agent to treat a wide variety of rheumatic diseases including psoriatic arthritis, juvenile arthritis, and discoid lupus erythematosus. Although the use of gold has been largely superseded by newer drugs, gold nanoparticles are being used effectively in laboratory based clinical diagnostic methods while concurrently showing great promise in vivo either as a diagnostic imaging agent or a therapeutic agent. For these reasons, gold nanoparticles are therefore well placed to enter mainstream clinical practice in the near future. Hence, the present review summarizes the chemistry, pharmacokinetics, biodistribution, metabolism, and toxicity of bulk gold in humans based on decades of clinical observation and experiments in which gold was used to treat patients with rheumatoid arthritis. The beneficial attributes of gold nanoparticles, such as their ease of synthesis, functionalization, and shape control are also highlighted demonstrating why gold nanoparticles are an attractive target for further development and optimization. The importance of controlling the size and shape of gold nanoparticles to minimize any potential toxic side effects is also discussed.

Potent, tumor-specific gene expression in an orthotopic hepatoma rat model using a Survivin-targeted, amplifiable adenoviral vector.

Ideal cancer gene therapies should have high tumor specificity and efficacy, and allow systemic administration to target metastases. We recently developed a bi-directional, two-step transcriptional amplification (TSTA) system driven by the tumor-specific Survivin promoter (pSurv) to amplify the correlated expression of both the reporter gene firefly luciferase (FL) and therapeutic gene tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Here, we compare the specificity and potency of an adenovirus carrying this system (Ad-pSurv-TSTA-TRAIL-FL) to a nonspecific vector (Ad-pCMV-FL) in an orthotopic hepatocellular carcinoma (HCC) rat model after systemic administration. At 24 h after injection of Ad-pCMV-FL, bioluminescence imaging revealed a trend (P=0.30) towards greater FL expression in liver versus tumor. In striking contrast, Ad-pSurv-TSTA-TRAIL-FL showed increased FL activity within the tumor compared with the liver (P<0.01), a strong trend towards reduced liver expression compared with Ad-pCMV-FL (P=0.07), and importantly, similar FL levels within tumor compared with Ad-pCMV-FL (P=0.32). Hence, this vector shows potent, tumor-specific transgene expression even after extensive liver transduction and may be of significant value in avoiding hepatotoxicity in HCC patients. Future studies will explore the benefits of tumor-specific TRAIL expression in this model, the potential to target metastases and the extension of this vector for the treatment of other Survivin-positive tumors is warranted.

Molecular imaging: current status and emerging strategies.

In vivo molecular imaging has a great potential to impact medicine by detecting diseases in early stages (screening), identifying extent of disease, selecting disease- and patient-specific treatment (personalized medicine), applying a directed or targeted therapy, and measuring molecular-specific effects of treatment. Current clinical molecular imaging approaches primarily use positron-emission tomography (PET) or single photon-emission computed tomography (SPECT)-based techniques. In ongoing preclinical research, novel molecular targets of different diseases are identified and, sophisticated and multifunctional contrast agents for imaging these molecular targets are developed along with new technologies and instrumentation for multi-modality molecular imaging. Contrast-enhanced molecular ultrasound (US) with molecularly-targeted contrast microbubbles is explored as a clinically translatable molecular imaging strategy for screening, diagnosing, and monitoring diseases at the molecular level. Optical imaging with fluorescent molecular probes and US imaging with molecularly-targeted microbubbles are attractive strategies as they provide real-time imaging, are relatively inexpensive, produce images with high spatial resolution, and do not involve exposure to ionizing irradiation. Raman spectroscopy/microscopy has emerged as a molecular optical imaging strategy for ultrasensitive detection of multiple biomolecules/biochemicals with both in vivo and ex vivo versatility. Photoacoustic imaging is a hybrid of optical and US techniques involving optically-excitable molecularly-targeted contrast agents and quantitative detection of resulting oscillatory contrast agent movement with US. Current preclinical findings and advances in instrumentation, such as endoscopes and microcatheters, suggest that these molecular imaging methods have numerous potential clinical applications and will be translated into clinical use in the near future.

Indirect imaging of cardiac-specific transgene expression using a bidirectional two-step transcriptional amplification strategy.

Transcriptional targeting for cardiac gene therapy is limited by the relatively weak activity of most cardiac-specific promoters. We have developed a bidirectional plasmid vector, which uses a two-step transcriptional amplification (TSTA) strategy to enhance the expression of two optical reporter genes, firefly luciferase (fluc) and Renilla luciferase (hrluc), driven by the cardiac troponin T (cTnT) promoter. The vector was characterized in vitro and in living mice using luminometry and bioluminescence imaging to assess its ability to mediate strong, correlated reporter gene expression in a cardiac cell line and the myocardium, while minimizing expression in non-cardiac cell lines and the liver. In vitro, the TSTA system significantly enhanced cTnT-mediated reporter gene expression with moderate preservation of cardiac specificity. After intramyocardial and hydrodynamic tail vein delivery of an hrluc-enhanced variant of the vector, long-term fluc expression was observed in the heart, but not in the liver. In both the cardiac cell line and the myocardium, fluc expression correlated well with hrluc expression. These results show the vector's ability to effectively amplify and couple transgene expression in a cardiac-specific manner. Further replacement of either reporter gene with a therapeutic gene should allow non-invasive imaging of targeted gene therapy in living subjects.

Evaluation of intracranial space-occupying lesion with Tc99m-glucoheptonate brain single photon emission computed tomography in treatment-naïve patients.

BACKGROUND: Glucoheptonate is a glucose analog with strong affinity for neoplastic brain tissues. Though extensively used as a tracer for detection of brain tumor recurrence, it's utility for characterization of intracranial lesions as neoplastic or otherwise has not been evaluated in treatment-naïve patients. AIM: The study was conducted to determine if glucoheptonate has sufficient specificity for neoplastic lesions of brain so that it can be utilized as a single photon emission computed tomography (SPECT)-tracer for differentiating neoplastic intracranial lesions from non-neoplastic ones in treatment-naïve patients. SETTINGS AND DESIGN: A cross-sectional analysis of treatment-naïve patients with intracranial space-occupying lesion done in a tertiary care hospital. MATERIALS AND METHODS: Fifty-four consecutive patients with clinical and radiological features of space-occupying lesion were included in this study. Glucoheptonate brain SPECT was performed before any definitive therapeutic intervention. Histopathological verification of diagnosis was obtained in all cases. STATISTICAL ANALYSIS USED: Descriptive statistics and student's 't' test. RESULT: Increased glucoheptonate uptake over the site of radiological lesion was noted in 41 patients and no uptake was noticed in 13 patients. Histopathology of 12 out of the 13 glucoheptonate non-avid lesions turned out to be non-neoplastic lesion; however, one lesion was reported as a Grade-2 astrocytoma. Histology from all the glucoheptonate concentrating lesions was of mitotic pathology. The sensitivity, specificity and accuracy of glucoheptonate for neoplastic lesion was 97.6%, 100% and 98.1%. CONCLUSIONS: Glucoheptonate has high degree of specificity for neoplastic tissues of brain and may be used as a tracer for SPECT study to differentiate neoplastic intracranial lesions from non-neoplastic ones.