In silico insights into design of novel VEGFR-2 inhibitors: SMILES-based QSAR modelling, and docking studies on substituted benzo-fused heteronuclear derivatives.

Eight QSAR models (M1-M8) were developed from a dataset of 118 benzo-fused heteronuclear derivatives targeting VEGFR-2 by Monte Carlo optimization method of CORALSEA 2023 software. Models were generated with hybrid optimal descriptors using both SMILES and Graphs with zero- and first-order Morgan extended connectivity index from a training set of 103 derivatives. All statistical parameters for model validation were within the prescribed limits, establishing the models to be robust and of excellent quality. Among all models, split-2 of M5 was the best-fit as reflected by rvalidation2, Qvalidation2 and MAE. Mechanistic interpretation of this model assisted the identification of structural descriptors as promoters and hinderers for VEGFR-2 inhibition. These descriptors were utilized to design novel VEGFR-2 inhibitors (YS01-YS07) by bringing modifications in compound MS90 in the dataset. Docking of all designed compounds, MS90 and sorafenib with VEGFR-2 binding site revealed favourable binding interactions. Docking score of YS07 was higher than that of MS90 and sorafenib. Molecular dynamics simulation study revealed sustained interactions of YS07 with key amino acids of VEGFR-2 at a run time of 100 ns. This study concludes the development of a best fit QSAR model which can assist the design of new anticancer agents targeting VEGFR-2.

Imaging-led symptomatic breast clinics: a case for direct access to radiology.

Radiologists play a pivotal role in patient management in modern hospital medicine and more so with regard to breast imaging. The diagnosis of breast pathology hinges predominantly on mammography and ultrasound imaging. With reduced clinician confidence in clinical examination alone, virtually all patients with breast symptoms are being referred for imaging. The traditional 'one-stop' clinics are victims of their own success and demand outstrips availability. This article makes a case for imaging-led breast clinics to increase efficiency, reduce duplication of work and cost, and increase throughput of patients.

Digital breast tomosynthesis within a symptomatic "one-stop breast clinic" for characterization of subtle findings.

OBJECTIVE: The aim of the study was to evaluate the diagnostic accuracy of combination of full-field digital mammography [two dimension (2D)] and digital breast tomosynthesis [DBT, three dimension (3D)] by comparing the combination with 2D imaging in a symptomatic setting. METHODS: A retrospective analysis was conducted involving 103 patients who attended symptomatic breast clinics between March 2012 and September 2012. All had subtle signs on 2D images or ultrasound. Mammographic score distribution was compared between 2D imaging and 2D + 3D imaging, followed by comparison with the gold-standard histopathology. Receiver operative characteristic curves and area under curve (AUC) were calculated for 2D imaging and the combination imaging (2D + 3D). SPSS(®) v. 21 (IBM Corp., New York, NY; formerly SPSS Inc., Chicago, IL) was used for data analysis with p < 0.05 as statistically significant. RESULTS: M3 lesions were reduced from 91 (85.8%) to 18 (16.9%) with the combination imaging. The mean AUC ± 95% confidence interval for 2D images alone was 0.721 (0.662-0.905) and for combined 2D and 3D images was 0.901 (0.765-1.00). The difference in AUCs between the two modalities was 0.180. CONCLUSION: DBT (3D imaging) increases diagnostic accuracy in a symptomatic breast clinic setting and reduces the number of M3 mammograms, when used as an adjuvant to 2D images. Therefore, DBT has the potential to increase workflow efficiency in a symptomatic setting by reducing benign biopsies. ADVANCES IN KNOWLEDGE: DBT reduces the number of M3 mammograms when used in the symptomatic breast setting and has the potential to reduce benign biopsies.

MRI texture analysis predicts p53 status in head and neck squamous cell carcinoma.

BACKGROUND AND PURPOSE: Head and neck cancer is common, and understanding the prognosis is an important part of patient management. In addition to the Tumor, Node, Metastasis staging system, tumor biomarkers are becoming more useful in understanding prognosis and directing treatment. We assessed whether MR imaging texture analysis would correctly classify oropharyngeal squamous cell carcinoma according to p53 status. MATERIALS AND METHODS: A cohort of 16 patients with oropharyngeal squamous cell carcinoma was prospectively evaluated by using standard clinical, histopathologic, and imaging techniques. Tumors were stained for p53 and scored by an anatomic pathologist. Regions of interest on MR imaging were selected by a neuroradiologist and then analyzed by using our 2D fast time-frequency transform tool. The quantified textures were assessed by using the subset-size forward-selection algorithm in the Waikato Environment for Knowledge Analysis. Features found to be significant were used to create a statistical model to predict p53 status. The model was tested by using a Bayesian network classifier with 10-fold stratified cross-validation. RESULTS: Feature selection identified 7 significant texture variables that were used in a predictive model. The resulting model predicted p53 status with 81.3% accuracy (P < .05). Cross-validation showed a moderate level of agreement (κ = 0.625). CONCLUSIONS: This study shows that MR imaging texture analysis correctly predicts p53 status in oropharyngeal squamous cell carcinoma with ∼80% accuracy. As our knowledge of and dependence on tumor biomarkers expand, MR imaging texture analysis warrants further study in oropharyngeal squamous cell carcinoma and other head and neck tumors.

How does semi-automated computer-derived CT measure of breast density compare with subjective assessments to assess mean glandular breast density, in patients with breast cancer?

OBJECTIVES: (a) To compare radiologists' breast mammographic density readings with CT subjective measures. (b) To correlate computer-derived measurement of CT density with subjective assessments. (c) To evaluate density distributions in this cohort of patients with breast cancer. METHODS: A retrospective review of mammograms and CT scans in 77 patients with breast cancer obtained within 1 year of each other was performed. Two radiologists independently reviewed both CT and mammograms and classified each case into four categories as defined by the breast imaging-reporting and data system of the American College of Radiology. Inter-reader agreements were obtained for both mammographic and CT density subjective evaluations by using the Cohen-weighted kappa statistic and Spearman correlation. The semi-automated computer-derived measurement of breast density was correlated with visual measurements. RESULTS: Inter-reader agreements were lower for subjective CT density grades than those for mammographic readings 0.428 [confidence interval (CI), 0.24-0.89] vs 0.571 (CI, 0.35-0.76). There was moderately good correlation between subjective CT density grades and the mammographic density grades for both readers (0.760 for Reader 1 and 0.913 for Reader 2). The semi-automated CT density measurement correlated well with the subjective assessments, with complete agreement of the density grades in 84.9% of patients and only one level difference in the rest. CONCLUSIONS: Semi-automated CT density measurements in the evaluation of breast density correlated well with subjective mammographic density measurement. ADVANCES IN KNOWLEDGE: There is good correlation between CT and mammographic density, but further studies are needed on how to incorporate semi-automated CT breast density measurement in the risk stratification of patients.

Online tuning of impedance matching circuit for long pulse inductively coupled plasma source operation--an alternate approach.

Impedance matching circuit between radio frequency (RF) generator and the plasma load, placed between them, determines the RF power transfer from RF generator to the plasma load. The impedance of plasma load depends on the plasma parameters through skin depth and plasma conductivity or resistivity. Therefore, for long pulse operation of inductively coupled plasmas, particularly for high power (∼100 kW or more) where plasma load condition may vary due to different reasons (e.g., pressure, power, and thermal), online tuning of impedance matching circuit is necessary through feedback. In fusion grade ion source operation, such online methodology through feedback is not present but offline remote tuning by adjusting the matching circuit capacitors and tuning the driving frequency of the RF generator between the ion source operation pulses is envisaged. The present model is an approach for remote impedance tuning methodology for long pulse operation and corresponding online impedance matching algorithm based on RF coil antenna current measurement or coil antenna calorimetric measurement may be useful in this regard.

Multiple delivery cesium oven system for negative ion sources.

Distribution of cesium in large negative ion beam sources to be operational in ITER, is presently based on the use of three or more cesium ovens, which operate simultaneously and are controlled remotely. However, use of multiple Cs ovens simultaneously is likely to pose difficulties in operation and maintenance of the ovens. An alternate method of Cs delivery, based on a single oven distribution system is proposed as one which could reduce the need of simultaneous operation of many ovens. A proof of principle experiment verifying the concept of a multinozzle distributor based Cs oven has been carried out at Institute for Plasma Research. It is also observed that the Cs flux is not controlled by Cs reservoir temperature after few hours of operation but by the temperature of the distributor which starts behaving as a Cs reservoir.

Screen-detected breast cancer: does presence of minimal signs on prior mammograms predict staging or grading of cancer?

AIM: To investigate whether the presence of minimal signs on prior mammograms predict staging or grading of cancer. MATERIALS AND METHODS: The previous mammograms of 148 consecutive patients with screen-detected breast cancer were examined. Women with an abnormality visible (minimal signs) on both current and prior mammograms formed the study group; the remaining patients formed the control group. Age, average size of tumour, tumour characteristic, histopathology, grade, and lymph node status were compared between the two groups, using Fisher's exact test. Cases in which earlier diagnosis would have made a significant prognostic difference were also evaluated. RESULTS: Eighteen percent of patients showed an abnormality at the site of the tumour on previous mammograms. There was no statistically significant difference between the two groups with respect to age, average size of tumour, histopathology, grade or lymph node status with p-values being 0.609, 0.781, 0.938, and 0.444, respectively. The only statistically significant difference between the two groups was tumour characteristics with more microcalcifications associated with either mass or asymmetrical density seen in the study group (p=0.003). Five patients in the study group showed lymph node positivity and were grade 3, and therefore, may have had possible gain from earlier diagnosis. CONCLUSION: The present study did not demonstrate a statistical difference in grading or staging between the group that showed "minimal signs" on prior mammograms versus normal prior mammograms. Microcalcification seems to be the most common characteristic seen in the missed cancer and a more aggressive management approach is suggested for breast microcalcifications.

Estimation of Total Phenols and Flavonoids in Extracts of Actaea spicata Roots and Antioxidant Activity Studies.

Actaea spicata Linn. (Ranunculaceae) has been traditionally used for the treatment of various ailments such as rheumatism, inflammation, nerve diseases, lumbago, scrofula and chorea, but no systematic phytochemical and pharmacological work has ever been carried out on this potential plant. Preliminary phytochemical screening showed presence of phenols and flavonoids in A. spicata. Thus, the present investigation was undertaken to estimate total phenols and flavonoids in methanol extract of A. spicata roots, and its ethyl acetate fraction. In vitro antioxidant activity was also evaluated in the methanol extract and ethyl acetate fraction using DPPH method. Ethyl acetate fraction was found to contain twice the content of flavonoids and phenols in comparison to methanolic extract, whereas phenolic content in methanol extract was approximately similar to ethyl acetate fraction. A significant antioxidant activity, i.e., mean percentage inhibition of DPPH radical was observed in methanol extract and ethyl acetate fraction at the concentration of 10 µg/ml and 5 µg/ml respectively. Finally, it was suggested that polyphenols are responsible for antioxidant activity of A. spicata.

Digital radiography. A comparison with modern conventional imaging.

The development of computed radiography over the past two decades has transformed radiological imaging. The radiology departments in the 21st century will look very different from those in the preceding period. In this review, the development of digital radiography is presented with a description of its various forms and a comparison with screen film radiography.

In vitro generation of cloned populations of Toxoplasma gondii.

Cloned populations were generated from Indian isolates of Toxoplasma gondii by transferring single tissue cysts from the brains of chronically infected mice to confluent murine macrophage (J774A.1) monolayers. The clones were then maintained continuously as tachyzoites in culture. Physical rupture of the tissue cysts and release of bradyzoites prior to seeding was found to be necessary for establishment of the parasite in culture. Although intact tissue cysts seeded over monolayers released bradyzoites spontaneously, they did not succeed in setting up an infection in the monolayers. Random amplified polymorphic DNA (RAPD)-PCR, which revealed distinct patterns for a clone and its progenitor, further confirmed the efficiency of the technique. The cloning technique was found to be simple and rapid compared to those involving limiting dilutions.

A rapid MTT colorimetric assay to assess the proliferative index of two Indian strains of Theileria annulata.

A study was undertaken to compare the proliferative index of macroschizont-infected lymphoblastoid cells of two Indian strains [Izatnagar (IZT) and Parbhani (PBN)] of Theileria annulata by an in vitro MTT [3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide], colorimetric assay. Culture conditions were standardized to define the optimal cell concentration in 96-well microculture plates to yield nearly 100% living cells for measurement of the metabolized formazan activity. A cell concentration of 1.5x10(5) cells/ml was found to be optimal for effective discrimination of the parasite strains. On the basis of conversion of MTT by the actively proliferating lymphoblastoid cells, the PBN strain of T. annulata stimulated a 2.5-fold increase in formazan activity in comparison to the IZT strain. The in vitro MTT assay was found to be a simple and convenient method for assessing the cell activation rate and growth, obviating the need for radioactive material for the assay. The results of the proliferation assay are discussed in relation to previously documented information on the biological characteristics of this important pathogen of cattle.

Evaluation of newer diagnostic methods for the detection and differentiation of Entamoeba histolytica in an endemic area.

We evaluated the colorimetric polymerase chain reaction (PCR)-based method for the detection and differentiation of Entamoeba spp. and compared the efficacy of E. histolytica-specific antigen detection in faeces with the detection of specific antibodies to E. histolytica-specific antigen in faeces, by enzyme-linked immunosorbent assay. Faecal samples were obtained from patients attending hospital in Chandigarh, India, from March 2001 to February 2002. The PCR-based colorimetric method was found to be the most sensitive (100%) and it could differentiate between pathogenic and non-pathogenic Entamoeba spp. The present study also emphasized that the antigen detection system may prove to be a better diagnostic tool than the antibody detection system in endemic areas.

Studies of apoptosis and bcl-2 in experimental atherosclerosis in rabbit and influence of selenium supplementation.

Apoptosis is a ubiquitous physiological mechanism of cell death regulating tissue mass and architecture. An attempt was made in the present study to see the occurrence of apoptotic cell death in three different treatment groups of rabbits viz. Control, HFD fed and HFD + Selenium fed. Apoptotic activity as checked by in situ DNA end labelling (TUNEL Assay) revealed excessive staining, mostly concentrated in plaque region both in fibrous as well as atheromatous plaque in HFD fed animals. However, in selenium (Se) supplemented animals, very little TUNEL staining could be seen, and even that confined to endothelial cells only. The control group on the other hand was totally devoid of any staining. Transmission Electron Microscopic (TEM) study also depicted the occurrence of apoptosis in aortic cells of HFD fed animals and very little in Se supplemented animals. Apoptotic activity has been discussed in relation to oxidative stress in HFD fed group. bcl-2, though an antiapoptotic oncoprotein, was found to be expressed more in atherogenic group as compared to control/HFD + Se treatment. On the whole, the study highlighted the occurrence of apoptotic process in atherosclerosis and the role of Se, a potent antioxidant, in inhibition of apoptotic process in HFD fed animals.

Giant-cell tumors of the patella.

This study describes the clinical, radiographic, and histopathologic features, and discusses the management options of 11 cases of benign giant-cell tumors of the patella seen over 20 years. Fine needle aspiration cytology was a useful preoperative diagnostic tool. Ptellectomy and reconstruction of the extensor apparatus is the favored treatment option with curettage and bone grafting restricted to selected localized lesions.

Influence of 7beta-hydroxycholesterol on glutathione status and nitric oxide production in macrophages: in vitro studies.

Incubation of murine peritoneal macrophages with 7beta-hydroxycholesterol (7beta-OH) for 24 hr led to dose-dependent reduction in cellular glutathione content as well as nitrite levels in the medium. Treatment with an inorganic form of selenium, sodium selenite which is a potent antioxidant, elevated the cellular glutathione levels and decreased nitrite levels. Our results suggest that 7beta-OH may exert its pro-atherogenic effect by inhibiting glutathione synthesis and nitric oxide production by macrophages present in the arterial wall and thus, impair the cellular antioxidant defense system.

Effect of 7beta-hydroxycholesterol on cellular redox status and heat shock protein 70 expression in macrophages.

The proatherogenic effects of oxidized low-density lipoproteins are attributed to oxysterols, particularly 7beta-hydroxycholesterol. In the present study, we have examined the effect of 7beta-hydroxycholesterol on cellular redox status and heat shock protein 70 (hsp70) expression. Murine peritoneal macrophages were incubated for 24 hours with increasing concentrations of 7beta-hydroxycholesterol (0-25 microg/ml). Intracellular reduced glutathione levels decreased and oxidized glutathione levels increased, thus increasing the redox ratio. Hsp70 expression in macrophages decreased in a dose-dependent fashion. It is suggested that 7beta-hydroxycholesterol exerts oxidative stress on macrophages and makes them more susceptible to its proatherogenic effects by impairing the cellular defense system consisting of antioxidants and stress proteins.

Detection of Theileria annulata carrier cattle by PCR.

A simple method for treating bovine blood samples for direct detection of T. annulata in carriers, after polymerase chain reaction (PCR) amplification of small subunit ribosomal RNA (SSU rRNA) gene is described. The threshold of detection of the PCR-assay was an erythrocytic parasitaemia of 0.00008% corresponding to 16 infected bovine erythrocytes. In 50 known carriers, 42 were positive in PCR, in which 8 cattle revealed presence of T. annulata in stained blood smear under microscope.

Safety and immunogenicity of recombinant Bacille Calmette-Guérin (rBCG) expressing Borrelia burgdorferi outer surface protein A (OspA) lipoprotein in adult volunteers: a candidate Lyme disease vaccine.

This phase I clinical trial was designed to determine the feasibility of using rBCG as a live bacterial vaccine vector for the outer surface protein A (OspA) of Borrelia burgdorferi and as model for other vaccines based on a rBCG vector. To construct the vaccine, a signal peptide derived from a mycobacterial lipoprotein was used to direct the export, and membrane-associated surface expression, of OspA in a standard strain of BCG (Connaught). The rBCG OspA vaccine was safe and immunogenic in several animal species, and protective in a mouse model of Lyme borreliosis. An intradermal injection (0.1 ml) of rBCG OspA was administered to 24 healthy adult volunteers sequentially at one of four dose levels, ranging from 2.0 x 10(4) CFU to 2 x 10(7) CFU, using a dose-escalation design. All volunteers were initially PPD-skin test and OspA antibody negative, and they were monitored for 2 years after immunization. Three volunteers had mild flu-like reactions 1-2 days after vaccination. Local ulceration and drainage at the site of injection, which occurred in 50% and 83% of volunteers in the two highest dose groups, persisted for 1-70 days before the ulcers healed. Most of the drainage samples yielded rBCG colonies that contained the OspA plasmid. Thirteen of 24 vaccinees, principally in the two highest dose groups, converted their PPD skin tests from negative to positive. None of the 24 volunteers developed OspA antibody. In conclusion, the current rBCG vaccine construct, the first such construct tested in humans, had a safety profile comparable to that of licensed BCG, but it did not elicit primary humoral responses to the vectored antigen.

Increased expression of fibroblast growth factor 8 in human breast cancer.

Fibroblast growth factor 8 (FGF8) is an important developmental protein which is oncogenic and able to cooperate with wnt-1 to produce mouse mammary carcinoma. The level of expression of FGF8 mRNA was measured in 68 breast cancers and 24 non-malignant breast tissues. Elevated levels of FGF8 mRNA were found in malignant compared to non-malignant breast tissues with significantly more malignant tissues expressing FGF8 (P=0.019) at significantly higher levels (P=0.031). In situ hybridization of breast cancer tissues and analysis of purified populations of normal epithelial cells and breast cancer cell lines showed that malignant epithelial cells expressed FGF8 mRNA at high levels compared to non-malignant epithelial and myoepithelial cells and fibroblasts. Although two of the receptors which FGF8 binds to (FGFR2-IIIc, FGFR3-IIIc) are not expressed in breast cancer cells, an autocrine activation loop is possible since expression of fibroblast growth factor receptor (FGFR) 4 and FGFR1 are retained in malignant epithelial cells. This is the first member of the FGF family to have increased expression in breast cancer and a potential autocrine role in its progression.