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Photoacoustic microscopy (PAM) leverages the optical absorption contrast of blood hemoglobin for high-resolution, multi-parametric imaging of the microvasculature in vivo. However, to quantify the blood flow speed, dense spatial sampling is required to assess blood flow-induced loss of correlation of sequentially acquired A-line signals, resulting in increased laser pulse repetition rate and consequently optical fluence. To address this issue, we have developed a sparse modeling approach for blood flow quantification based on downsampled PAM data. Evaluation of its performance both in vitro and in vivo shows that this sparse modeling method can accurately recover the substantially downsampled data (up to 8 times) for correlation-based blood flow analysis, with a relative error of 12.7 ± 6.1 % across 10 datasets in vitro and 12.7 ± 12.1 % in vivo for data downsampled 8 times. Reconstruction with the proposed method is on par with recovery using compressive sensing, which exhibits an error of 12.0 ± 7.9 % in vitro and 33.86 ± 26.18 % in vivo for data downsampled 8 times. Both methods outperform bicubic interpolation, which shows an error of 15.95 ± 9.85 % in vitro and 110.7 ± 87.1 % in vivo for data downsampled 8 times.

Photo acoustic microscopy (PAM) achieves high contrast, intravital imaging of the microvasculature by utilizing the specificity of endogenous optical absorption. PAM has been further augmented by using loss of correlation (LoC) methods to image blood flow. However, estimating blood flow using LoC methods necessitates dense spatial sampling which increases laser fluence. To address concerns over the increase in laser fluence, we develop a sparse modeling algorithm to reconstruct blood flow in PAM from downsampled data. The proposed method is superior to reconstruction by bicubic interpolation, exhibiting an error of 5.6 ± 3.4% in vivo compared to 33.4 ± 32.7% for bicubic interpolation, for data downsampled eight times.

Conventionally echocardiography uses plane or diverging transmit beams to monitor fast movements of hearts. However, reduced sensitivity and increased near field artifacts limits their applications in deep tissue. Tissue harmonic imaging, using focused ultrasound beams, is capable of improving image quality but is associated with a lower frame rate. In this study, we demonstrate the combination of hierarchical compressed sensing with tissue harmonic imaging enables increase in the frame rate. With 50%, 33.3% and 25% of original A-line number, our proposed method suppressed root mean squared error to 6.3%, 7.6% and 8.3%, respectively. In addition, the method was validated with a custom imaging sequence in vivo.

Nearly 80% of cancer patients do not have genetic mutation results available at initial oncology consultation; up to 25% of patients begin treatment before receiving their results. These factors hinder the ability to pursue optimal treatment strategies. This study validates a blood-based genome-testing service that provides accurate results within 72 hours. We focused on targetable variants in advanced non-small cell lung carcinoma-epidermal growth factor receptor gene (EGFR) variant L858R, exon 19 deletion (ΔE746-A750), and T790M; GTPase Kirsten ras gene (KRAS) variants G12C/D/V; and echinoderm microtubule associated protein like and 4 anaplastic lymphoma receptor tyrosine kinase fusion (EML4-ALK) transcripts 1/2/3. Test development included method and clinical validation using samples from donors with (n = 219) or without (n = 30) cancer. Clinical sensitivity and specificity for each variant ranged from 78.6% to 100% and 94.2% to 100%, respectively. We also report on 1643 non-small cell lung carcinoma samples processed in our CLIA-certified laboratory. Mutation results were available within 72 hours for 94% of the tests evaluated. We detected 10.5% mutations for EGFR sensitizing (n = 2801 samples tested), 13.8% mutations for EGFR resistance (n = 1055), 13.2% mutations in KRAS (n = 3477), and 2% mutations for EML4-ALK fusion (n = 304). This rapid, highly sensitive, and actionable blood-based assay service expands testing options and supports faster treatment decisions.

Photoacoustic microscopy acquires volumetric RF data to obtain high resolution, high contrast, images of the microvasculature but is associated with slow acquisition of data due to mechanical raster scanning across the image plane. Recent work has shown that the acquisition speed can be increased using compressive sampling methods and subsequent reconstruction. These methods use bases (dictionaries) learned from prior fully sampled acquisitions, or classical bases such as the Fourier or wavelet bases. In this study, we present the simultaneous learning of bases, and reconstruction using only subsampled data. The algorithm was validated at two different subsampling levels 50% and 75% downsampling, and compared to the ground truth reconstruction with fully sampled data by estimating the peak signal to noise ratio (PSNR). No significant difference in performance was observed between the fully sampled (20.0±3.0 dB), 50% (19.9±2.1 dB) and 75% (19.1±2.6 dB) subsampled data.

Dictionary learning is an unsupervised learning method to abstract image data into a set of learned basis vectors. In prior work, the efficacy of the K-SVD dictionary learning algorithm in suppressing reverberation in volumetric photoacoustic microscopy (PAM) data was demonstrated. In this work, we compare the K-SVD algorithm against the method of optimal directions (MOD). The generalization error and reverberation suppression performance of the two algorithms were compared. The K-SVD was found to have a lower average generalization error (5.69x104 ±9.09x103 (a.u.)) when compared to the MOD (8.27x104 ±1.33x104 (a.u.)) for identical training data, initialization, sparsity (3 atoms per A-line) and number of iterations (5). Both algorithms were observed to suppress the reverberation to a similar extent (18.8 ± 1.12 dB for the K-SVD and 18.3 ± 1.2 dB for the MOD). Our data show that irrespective of the method used, sparse dictionary learning can significantly suppress reverberations in PAM.

9 p., Background: Stroke disproportionately affects people in low-income and middle-income countries. Although improvements in stroke care and outcomes have been reported in high-income countries, little is known about practice and outcomes in low and middle-income countries. We aimed to compare patterns of care available and their association with patient outcomes across countries at different economic levels. Methods: We studied the patterns and effect of practice variations (ie, treatments used and access to services) among participants in the INTERSTROKE study, an international observational study that enrolled 13 447 stroke patients from 142 clinical sites in 32 countries between Jan 11, 2007, and Aug 8, 2015. We supplemented patient data with a questionnaire about health-care and stroke service facilities at all participating hospitals. Using univariate and multivariate regression analyses to account for patient casemix and service clustering, we estimated the association between services available, treatments given, and patient outcomes (death or dependency) at 1 month. Findings: We obtained full information for 12 342 (92%) of 13 447 INTERSTROKE patients, from 108 hospitals in 28 countries; 2576 from 38 hospitals in ten high-income countries and 9766 from 70 hospitals in 18 low and middle-income countries. Patients in low-income and middle-income countries more often had severe strokes, intracerebral haemorrhage, poorer access to services, and used fewer investigations and treatments (p

Echocardiographic image sequences are frequently corrupted by quasi-static artifacts known as “clutter”. These artifacts are superimposed on the moving myocardium and hinder useful diagnosis. Prior work has shown the efficacy of blind source separation methods to suppress clutter while retaining underlying signal. However, the same image may be corrupted with clutter from multiple mechanisms or sources. Consequently, the spatiotemporal characteristics of the clutter vary depending on the mechanism. In this paper, a method is proposed that progressively detects and removes clutter of increasing temporal rank using sparse coding. The performance was quantified in vitro by measuring the mean squared error with respect to a ground truth dataset with no clutter. The error with respect to the ground truth was reduced from 1.3×103 ± 4.7×102 a.u. to 2.2 × 10−2 ± 9.8×10−3 a.u. The average clutter reduction in vivo was 11.9±0.5 dB.

Photoacoustic Microscopy (PAM) offers a unique combination of high contrast from endogenous optical absorbers, and increased penetration to image microvasculature. However, images of the vasculature at increased depth are often corrupted by acoustic reverberation from superficial layers. In this paper, we present an algorithm using dictionary learning to remove the reverberant signal while preserving underlying microvascular anatomy. The algorithm was validated in vitro, using dyed beads embedded polydimenthylsiloxane (PDMS). Subsequently, we demonstrate suppression of reverberant artifacts by 20 ± 0.2 dB using in vivo PAM data acquired in a mouse brain.

It is recommended to utilize a dynamic multithreaded OS for WSN (Wireless Sensor Network) platforms. This operating system gives a helpful threading model, for Underwater Sensor network applications. Virtual reminiscence is bolstered by means of the collaboration of the compiler, consequently that the sensor network policies is to be able to implement code bigger than the corporeal code reminiscence they have. In the direction of empower firmware upgrade intended for sent sensor nodes, introduced operating system hold ups wireless reinventing. The proposed techniques don't build hardware price, along with are intended in the direction of oblige little progressions towards active functions. It has been created a compression algorithm appropriate to this application. The normally utilized library as well as the major legitimate configuration is isolated; every sensor system has a duplicate of the active File structure library inside the MMC (Multi Media Card), as well as consequently just the basic task along with user characterized subroutines can be updated through RF. A dynamic, proficient file system named Flash File System is likewise contained within OS. The code size as well as data size of OS including Flash File system is 8kB and 646 bytes, individually.

Echocardiographic image sequences are frequently corrupted by quasi-static “clutter” artifacts superimposed on the dynamic myocardium. Blind Source Separation (BSS) aIgorithms are often used to suppress clutter while retaining underlying tissue. In this paper, the sparse nature of reverberant artifacts in the time frequency domain is exploited for suppression to achieve reduced computational complexity and tissue signal loss. The performance of the method was validated on five datasets simulated in Field-II and three in vivo datasets and benchmarked with respect to the Singular Value Filter (SVF) algorithm. In simulated data, the proposed method yields a normalized average error of 1.15±0.09 a.u. over five simulated datasets, compared to the SVF, which yields an error of 14.13±1.98 a.u. on the same datasets. The suppressed signal energy was also estimated to quantify loss of ‘Tissue signal’. The average energy of the suppressed signal for the SVF was 35.60±3.27 a.u. The average energy of the suppressed signal of the proposed method was 25.44±1.95 a.u. Incorporating sparsity and signal characteristics allowed us to accelerate the clutter suppression by three orders of magnitude in this case.

Prostate cancer is a clinically heterogeneous and multifocal disease. More than 80% of patients with prostate cancer harbor multiple geographically discrete cancer foci at the time of diagnosis. Emerging data suggest that these foci are molecularly distinct consistent with the hypothesis that they arise as independent clones. One of the strongest arguments is the heterogeneity observed in the status of E26 transformation specific (ETS) rearrangements between discrete tumor foci. The clonal evolution of individual prostate cancer foci based on recent studies demonstrates intertumoral heterogeneity with intratumoral homogeneity. The issue of multifocality and interfocal heterogeneity is important and has not been fully elucidated due to lack of the systematic evaluation of ETS rearrangements in multiple tumor sites. The current study investigates the frequency of multiple gene rearrangements within the same focus and between different cancer foci. Fluorescence in situ hybridization (FISH) assays were designed to detect the four most common recurrent ETS gene rearrangements. In a cohort of 88 men with localized prostate cancer, we found ERG, ETV1, and ETV5 rearrangements in 51% (44/86), 6% (5/85), and 1% (1/86), respectively. None of the cases demonstrated ETV4 rearrangements. Mutual exclusiveness of ETS rearrangements was observed in the majority of cases; however, in six cases, we discovered multiple ETS or 5′ fusion partner rearrangements within the same tumor focus. In conclusion, we provide further evidence for prostate cancer tumor heterogeneity with the identification of multiple concurrent gene rearrangements.

Existing clinical imaging procedures lack sensitivity and specificity in detecting early prostate cancer bone me- tastatic lesions. In this study, we developed a highly reproducible bone metastasis xenograft model and identified possible molecular imaging candidates for detecting early bone metastatic lesions. Bone trophic human prostate cells (PC-3B1) were isolated and characterized for their ability to reach bone after intracardiac injection into SCID mice. The appearances of skeletal metastases were evaluated using digital radiographic imaging and confirmed by necropsy and histology. The PC-3B1 cells retain a bone homing phenotype after long term propagation in tissue culture and exhibit progressive bone lesions within 3 weeks following intracardiac injection. Comparative transcription signatures of PC-3 and PC-3B1 cells were determined using a cancer specific microarray and confirmed by RT-PCR analysis. The analysis identified increased expression of four cell surface molecules in PC-3B1 cells that may be suitable as molecular imaging candidates to detect bone micro metastases.

We investigated the aberrant promoter methylation status of 12 genes in skin lesions, both malignant (basal cell carcinomas (BCCs), n=68 and squamous cell carcinomas (SCCs), n=35) and non-malignant (tags, n=58) skin lesions and compared the results of lesions from sun exposed (SE) and sun protected (SP) regions. Methylation was studied using a methylation specific PCR (MSP) and methylation of CDH1 was also measured using a semi-quantitative fluorescence based real-time MSP method. The methylation index (MI) was calculated as the methylated fraction of the genes examined. In this report, we found high frequencies of methylation of several known or suspected tumor suppressor genes in tags and skin cancers. Among the 12 genes, for the cadherin genes CDH1 and CDH3 and for two of the laminin 5 encoding genes LAMA3 and LAMC2 methylation frequencies greater than 30% were noted in one or more specimen types. We investigated whether methylation was tumor related. Surprisingly, the differences in the methylation profile of genes among the three specimen types were modest, and the MI, indicators of overall methylation frequencies, was nearly identical. However, significant differences were noted in the frequencies of methylation among the three specimen types for the genes RASSF1A (P=0.002), CDH1 (P=0.007) and one or more of three CAD genes (P=0.02). Methylation was highly significantly related to sun exposure, and sun protected specimens had little or no methylation. As methylation of CDH1 was completely SE specific we analyzed all the skin samples using a semi-quantitative real-time PCR assay for the CDH1 gene. The concordance between standard MSP and real-time MSP for all the samples (n=161) was 75% (P

Watery extracts of forty plant species commonly growing in across India have been screened for antisporulant activity against Sclerospora graminicola (Sacc.) Schroet., the causative agent of pearl millet downy mildew. The collection represented 38 genera of 30 families. The extracts of thirteen species did not show any effect, whereas the activity of extracts of Allium sativum, Clematis gouriana, Evolvulus alsinoides, Mimusops elengi, Parthenium hysterophorus, Piper nigrum and Tagetes erecta were commensurable to that of marketed botanical fungicides and Mikal 70 wp. The crude extracts of 12 species (Agave americana, Aloe vera, Artemisia parviflora, Citrus limon, Citrus sinensis, Eucalyptus globosus, Euphorbia hirta, Leucas aspera, Murraya koenigi, Ocimum sanctum, Santalum album and Zingiber offinale) completely inhibited the zoosprorangium formation while in the case of remaining 8 plants the crude extracts reduced only partially the sporulation. The antisporulant activity of commercialised Azadirachta preparation (Nutri-Neem) was more pronounced than that of Reynutria based one (Milsana) and Sabadilla (veratrin), however, these botanical preparations held off synthetic fungicides and the most active watery extracts.

In this paper, we present whole-organ histologic and genetic mapping studies using hypervariable DNA markers on chromosome 13 and then integrate the recombination- and single-nucleotide polymorphic sites (SNPs)-based deletion maps with the annotated genome sequence. Using bladders resected from patients with invasive urothelial carcinoma, we studied allelic patterns of 40 microsatellite markers mapping to all regions of chromosome 13 and 79 SNPs located within the 13q14 region containing the RB1 gene. A whole-organ histologic and genetic mapping strategy was used to identify the evolution of allelic losses on chromosome 13 during the progression of bladder neoplasia. Markers mapping to chromosomal regions involved in clonal expansion of preneoplastic intraurothelial lesions were subsequently tested in 25 tumors and 21 voided urine samples of patients with bladder cancer. Four clusters of allelic losses mapping to distinct regions of chromosome 13 were identified. Markers mapping to the 13q14 region that is flanked by D13S263 and D13S276, which contains the RB1 gene, showed allelic losses associated with early clonal expansion of intraurothelial neoplasia. Such losses could be identified in approximately 32% bladder tumor tissue samples and 38% of voided urines from patients with bladder cancer. The integration of distribution patterns of clonal allelic losses revealed by the microsatellite markers with those obtained by genotyping of SNPs disclosed that the loss within an approximately 4-Mb segment centered around RB1 may represent an incipient event in bladder neoplasia. However, the inactivation of RB1 occurred later and was associated with the onset of severe dysplasia/carcinoma in situ. Our studies provide evidence for the presence of critical alternative candidate genes mapping to the 13q14 region that are involved in clonal expansion of neoplasia within the bladder antecedent to the inactivation of the RB1 gene.

In the present study, Dravya ‐ an organic compound used for seed treatment along with the common fungicides to test its compatibility in the management of Alternaria padwickii in paddy. Dravya (a sea weed extract) was found highly compatible with fungicides like Bavistin and Dithane M‐45. Incidence of Alternaria padwickii and Bipolaris oryzae was also reduced to a greater extent in the paddy seed samples of Dravya treatment. On the other hand, it also enhanced the seed germination and seedling vigour. Seedlings of treated samples also showed enhanced activity of peroxidase upon challenge inoculation with Alternaria padwickii. The enzyme activity in the seedlings challenged with the pathogen was two fold more over control. The suppression in disease incidence in growing plants indicated the promising effect of Dravya and Dithane M‐45 under green‐ house conditions.

In order to trigger defense mechanisms in plants, abiotic or biotic factors can be used as inducers/elicitors. In the present study, Dravya (a sea weed extract) was evaluated for its compatibility with common synthetic fungicides like Bavistin and Dithane M-45. Dravya was found to be highly compatible with Dithane M-45, in which the incidence of Bipolaris oryzae and Alternaria padwickii was reduced to a greater extent in the paddy seed sample of Cv. IR-64, a popular variety for popped rice in South India. In parallel, seed germination and seedling vigour were also enhanced over control. Treated seedlings also indicated the enhanced peroxidase and phenylalanine ammonia-lyase activities upon challenge inoculation with Bipolaris oryzae. Challenged seedlings showed higher activity of enzymes on second and fourth day after inoculation. The suppression in disease incidence of the seedlings was also noticed in the growing plants indicated the promising effects of Dravya with Dithane M-45 under green house conditions.

Deregulation of cell cycle inhibition contributes to human carcinogenesis. Reprimo (for stop/repress) is a newly identified mediator of the p53-mediated cell cycle arrest at the G2 phase. Loss of Reprimo expression due to promoter methylation was recently identified in pancreatic cancer. We examined Reprimo expression by reverse transcription PCR (RT-PCR) and aberrant methylation of Reprimo by methylation specific PCR (MSP) in lung cancer cell lines (n=35) and primary tumors (n=167). We also correlated the p53 gene status with Reprimo methylation in cell lines. Aberrant methylation of Reprimo was present in 32% (six of 19) of non-small cell lung cancer (NSCLC) cell lines, 6% (one of 16) of small cell lung cancer (SCLC) cell lines, and 31% (51 of 167) of primary tumors. Methylation was absent in normal lymphocytes and was rare in corresponding nonmalignant lung tissues (7%; four of 57). Overall concordance between loss of expression and aberrant methylation of Reprimo was 94% (33 of 35) in cell lines. Reprimo expression was restored after treatment with the demethylating agent 5-aza-2'-deoxycytidine in all five-cell lines tested that lacked Reprimo expression. There was no significant correlation between p53 gene status and Reprimo methylation in cell lines. These data indicate that Reprimo methylation is frequent in lung cancers and occurs independently of p53 status. Methylation of Reprimo may play a role in the pathogenesis of lung cancers.

Methanolic extracts of forty plant species commonly growing across India were collected and have been screened for antisporulant activity against Sclerospora graminicola (Sacc.) Schroet., the causative agent of pearl millet downy mildew. The collection represented 38 genera of 30 families. The methanolic extracts of nine species did not show any effect, whereas the activity of the extracts of Clematis gouriana, Evolvulus alsinoides, Mimusops elengi, Allium sativum and Piper nigrum were commensurable to that of the marketed botanical fungicides. The extracts of 11 species (Agave americana, Artemisia pallens, Citrus sinensis, Dalbergia latifolia, Helianthus annus, Murraya koenigii, Ocimum basilicum, Parthenium hysterophorus, Tagetes erecta, Thuja occidentalis and Zingiber offinale) exhibited remarkable antisporulant effect even after 10-fold dilution of the crude extracts while in the case of remaining 15 plants the crude extracts loosed activity after 10-fold dilution. The antisporulant activity of commercialised Azadirachta preparation (Nutri-Neem) was more pronounced than that of Reynutria based one (Milsana) and Sabadilla (veratrin), however, these botanical preparations held off the extracts of C. gouriana and E. alsinoides and synthetic fungicides.

Despite major improvements in patient management, the prognosis for patients with lung cancer remains dismal. As our knowledge of the molecular biology of cancers has increased, new targets for therapeutic interventions have been identified. In this article, we discuss some of the more recent developments in this field. They include revisiting some of the established concepts, such as retinoid metabolism and the inhibition of cyclooxygenase-2 metabolism. In addition, newer targets, such as transforming growth factor-beta signaling, Janus-activated kinase/signal transducers and activators of transcription pathway, and cell invasion are discussed. These studies demonstrate that multiple, often overlapping, mechanisms of disruption are present in lung cancer cells, presenting a plethora of molecular targets.

Laminin-5 (LN5) anchors epithelial cells to the underlying basement membrane, and it is encoded by three distinct genes: LAMA3, LAMB3, and LAMC2. To metastasize and grow, cancer cells must invade and destroy the basement membrane. Our previous work has shown that epigenetic inactivation is a major mechanism of silencing LN5 genes in lung cancers. We extended our methylation studies to resected bladder tumors (n = 128) and exfoliated cell samples (bladder washes and voided urine; n = 71) and correlated the data with clinicopathologic findings. Nonmalignant urothelium had uniform expression of LN5 genes and lacked methylation. The methylation frequencies for LN5 genes in tumors were 21–45%, and there was excellent concordance between methylation in tumors and corresponding exfoliated cells. Methylation of LAMA3 and LAMB3 and the methylation index were correlated significantly with several parameters of poor prognosis (tumor grade, growth pattern, muscle invasion, tumor stage, and ploidy pattern), whereas methylation of LAMC2 and methylation index were associated with shortened patient survival. Of particular interest, methylation frequencies of LAMA3 helped to distinguish invasive (72%) from noninvasive (12%) tumors. These results suggest that methylation of LN5 genes has potential clinical applications in bladder cancers.

HIN-1 (high in normal-1) is a putative cytokine with growth inhibitory activities and is downregulated by aberrant methylation in breast cancers. We studied HIN-1 methylation status in many types of adult and pediatric malignancies and cell lines. We examined the expression of HIN-1 mRNA in 52 cell lines and the promoter methylation status in the cell lines and in over 800 primary tumors representing 17 tumor types using methylation specific PCR. Promoter methylation was observed in 73% of breast cancer, 67% of nonsmall cell lung cancer (NSCLC), 30% of small cell lung cancer (SCLC) and 57% of malignant mesothelioma (MM) cell lines, and methylation was completely correlated with loss of expression. Expression negative cell lines restored HIN-1 expression after treatment with 5-aza-2'-deoxycytidine. Promoter methylation of HIN-1 was found in 90% of retinoblastomas, 73% of Wilms' tumors, 61% of rhabdomyosarcomas, 57% of breast cancers, 52% of prostate cancers, 40% of MMs, 28% of NSCLCs and 27% of lymphomas. Methylation frequencies in colorectal cancers, cervical cancers, bronchial carcinoids, SCLCs, neuroblastomas, osteosarcomas, leukemia, medulloblastomas and bladder cancers were lower (4-21%), while hepatoblastomas lacked methylation. HIN-1 methylation was rarely detected in nonmalignant tissues (8 of 165, 5%). Aberrant methylation of HIN-1 with loss of expression is a common event and may contribute to the pathogenesis of many types of human malignancies.

We have previously reported that the key apoptosis related gene caspase 8 (CASP8) is frequently silenced in small cell lung cancer (SCLC) tumors and cell lines usually, but not always, by aberrant promoter methylation. Because CASP8 is a key component of the death-inducing signaling complex (DISC) when specific death receptors (including DR4, DR5, FAS) are activated by their specific ligands (TRAIL/FASL), we examined expression of the components of the DISC complex in lung cancer cell lines. MYC family members are frequently amplified (MYC+ve) in SCLC, and MYC is a potent inducer of apoptosis. We examined 34 SCLC lines (12 of which were MYC+ve) and 22 NSCLC lines. CASP8 gene expression was frequently lost (79%) at message and protein levels in SCLC but not in non-SCLC (NSCLC). MYC amplification was present in 45% of SCLC cell lines, which had lost CASP8 expression, but not in any of the CASP8 positive lines. The frequency of CASP8 loss was significantly higher in MYC+ve SCLC compared to MYC-ve SCLC or in NSCLC. Analyses of other DISC components showed significantly higher rates of loss of expression of CASP10, DR5, FAS and FASL in SCLC compared to NSCLC. The loss of expression of proapoptotic DISC components was significantly higher in MYC+ve SCLC cell lines and these lines were completely resistant to TRAIL. Expression of CASP10 (a caspase closely related to CASP8) was frequently absent at the protein level in both SCLC and NSCLC lines. Expression of c-FLIP (proteolytically inactive homolog of CASP8) was inversely related to expression of CASP8. Our major conclusions are: (a) The death receptor pathway is differently inactivated at multiple levels in lung cancer cell lines; and (b) MYC amplification in SCLC is associated with inactivation of most components of the DISC complex, with resistance to TRAIL and with expression of c-FLIP. These findings may have considerable clinical and therapeutic implications.

The nucleosomal chromatin structure within genes is disrupted upon transcription by RNA polymerase II. To determine whether this disruption is caused by transcription per se as opposed to the RNA polymerase source, we engineered the yeast chromosomal HSP82 gene to be exclusively transcribed by bacteriophage T7 RNA polymerase in vivo. Interestingly, we found that a fraction of the T7-generated transcripts were 3' end processed and polyadenylated at or near the 3' ends of the hsp82 and the immediately downstream CIN2 genes. Surprisingly, the nucleosomal structure of the T7-transcribed hsp82 gene remained intact, in marked contrast to the disrupted structure generated by much weaker, basal level transcription of the wild type gene by RNA polymerase II under non-heat shock conditions. Therefore, disruption of chromatin structure by transcription is dependent on the RNA polymerase source. We propose that the observed RNA polymerase dependence for transcription-induced nucleosome disruption may be related either to the differential recruitment of chromatin remodeling complexes, the rates of histone octamer translocation and nucleosome reformation during polymerase traversal, and/or the degree of transient torsional stress generated by the elongating polymerase.

During the manufacture of austenitic stainless steel hypodermic needle tubes by sinking operation, unacceptable inner surface roughness was developed. By analysing the process conditions the cause was attributed to the uncontrolled plastic flow of the material during the reduction process. By using a moving mandrel or introducing an intermediate drawing operation with suitable mandrel this could be controlled to get the acceptable quality product.

Using a genetic strategy designed to find proteins involved in the function of the Saccharomyces cerevisiae transcriptional activator GAL4, we isolated mutants in two genes which rescue a class of gal4 activation domain mutants. One of these genes, SUG1, encodes a member of a large family of putative ATPases, the Conserved ATPase containing Domain (CAD) proteins (also known as AAA proteins) that are involved in a wide variety of cellular functions. Subsequently, SUG1 was identified as a subunit of the 26 S proteasome. We have now cloned the gene defined by the second complementation group. SUG2 encodes an essential 49-kDa protein that is also a member of the CAD family and is 43% identical to SUG1. The mutation in sug2-1, like that in sug1-1, is found in the CAD near the highly conserved ATPase motif. We present biochemical and genetic evidence that SUG2 is associated in vivo with SUG1 and is a novel CAD protein subunit of the 26 S proteasome. With its highly conserved mammalian homologs, human p42 and ground squirrel CADp44, SUG2 defines a new class of proteasomal CAD proteins.

The failure analysis of prematurely failed chassis beam of a light commercial vehicle was carried out. The metallographic examination of the material indicated high levels of slag (sulphide and silicate) inclusions and uneven distribution of ferrite and pearlite phases. The failure has essentially taken place through the bolt holes. The analysis suggested that inferior quality material led to the failure, whereas the bolt holes might have acted only as stress concentration centres for the initiation of the cracks.

The intersegmental muscles (ISMs) of the tobacco hawkmoth Manduca sexta participate in the emergence behavior of the adult moth at the end of metamorphosis and then die during the subsequent 30-hr period. The trigger for this death is a decline in the circulating titer of the insect molting hormone 20-hydroxyecdysone (20-HE). Previous work has demonstrated that the ability of the ISMs to die is dependent on new gene expression. Using a differential hybridization cloning strategy, a cDNA library made from the ISMs committed to die was screened, and four up-regulated clones were isolated. One clone, 18-56, was selected for this study. Northern and Western analysis demonstrated that while clone 18-56 was expressed in all tissues examined and during every stage of ISM development, there was a dramatic increase in expression at both mRNA and protein levels when the ISMs became committed to die. If ISM death was delayed by an injection of 20-HE on the day proceeding adult emergence, 18-56 expression remained at basal levels. Immunocytochemistry demonstrated that 18-56 protein was located predominantly in nuclei prior to the commitment of the ISMs to die and then accumulated to high levels in cytoplasm at the time of cell death. DNA sequence analysis revealed that 18-56 protein shares 74% identity with yeast SUG1 and 92% with human Trip1, both of which are members of the conserved CAD (Conserved ATPase-containing Domain) family of putative transcriptional regulators. To verify that these genes shared functional as well as sequence homology, Manduca clone 18-56 was transformed into a yeast mutant for SUG1 function. Manduca 18-56 was able to both complement the lethal SUG1 phenotype and to suppress the transcriptional activity of a SUG1 mutation in yeast. Taken together, these data support the hypothesis that members of the phylogenetically conserved CAD family participate in important basal and developmental processes.

The insecticidal crystal protein gene cryIA(a) from Bacillus thuringiensis HD1 has been cloned as a single 3.765 kb Ndel fragment on the expression vector pRE1. The pBR322 based clone pES1 was digested with restriction endonuclease Ndel and the 3.765 kb fragment carrying the intact gene was eluted and cloned on pUC18 to confirm its functional integrity. This Ndel fragment was then cloned on the vector pRE1 carrying strong promoter PL of lambda upstream to the cloning site. The recombinant construct pUSR14.1 carried crystal protein (CP) gene under PL and was temperature inducible at 42°C in MZ1 host strain of Escherichia coli because of temperature sensitive CI857 gene carried by it as lysogen. Dilution based insect bioassays showed hyper-expression of toxin in these constructs. SDS-PAGE analysis indicated that polypeptides corresponding to 132 kD and 66 kD bands of HD1 endotoxin constituted 20.1% of the total soluble protein in this recombinant strain to be delta-endotoxin.

A rare case of gastric duplication cyst of tongue is reported.

Investigations were carried out on the oxidation characteristics of artificially layered Fe/Al and Fe/Mg thin films using X-ray diffraction and Raman spectroscopy at 303, 473, 673 and 773 K in the laboratory atmosphere. X-ray diffractograms and Raman spectra reveal diffusion phenomena occurring in Fe/Al thin films. Individual oxides and a spinel-phase MgFe2O4, were the oxidation products in Fe/Mg multilayers while intermetallics were found to be present in the Fe/Al system at 773 K, in addition to FeAl2O4.

BACKGROUND: Finger millet is a major food crop as well as feed and fodder for livestock, especially in regions of southern India. A sturdy crop to fluctuating environmental conditions, it can be cultivated in all seasons of the year. Leaf, neck and finger blast caused by Pyricularia grisea Sacc. and Bipolaris setariae (Saw.) Shoem, as well as leaf spot disease, Bipolaris nodulosa (Berk & M.A.Curtis) Shoem, are major production constraints in southern India. Apart from environmental conditions, the use of harvested seeds by farmers is a major reason for disease prevalence. Benzophenone analogues have been investigated for controlling phytopathogenic fungi. In addition, the most important applications of azetidin-2-ones are as antibiotics. Based on this information, the present study was conducted to explore the antifungal activity of integrated 2-azetidinonyl and 1,3,4-oxadiazoles moieties into a benzophenone framework. RESULTS: A simple high-yielding method for the integration of heterocyclic rings, namely 2-azetidinonyl, at the benzophenone nucleus has been achieved, starting from substituted 2-hydroxybenzophenones under mild conditions on a wet solid surface using microwave irradiation. In the present study, an array of newly synthesised compounds, 2-azetidinonyl-5-(2-benzoylphenoxy)methyl-1,3,4-oxadiazoles, were screened for their antifungal property against blast and leaf spot causing fungi associated with the seeds of finger millet, cv. Indof-9. CONCLUSION: Two of the newly synthesised compounds showed promising effects in depleting the incidence of seed-borne pathogenic fungi of finger millet. The suppression of Pyricularia grisea and Bipolaris setariae resulted in enhanced seed germination and seedling growth. (C) 2009 Society of Chemical Industry

A multi-marked strain ofRhizobium meliloti was developed by the co-mutation method and employed to contribute to the genetic map ofR. meliloti chromosome. Seven loci were placed at 5 sites in theleu region in the orderman-aba, fix, leu-cro-azt, ost-thi.

Oxidation kinetics of ferrochromium (Cr 72 wt%, Fe 28 wt%) films of different thicknesses were followed at room temperature in the laboratory atmosphere by noting the conductivity of vacuum-deposited films as a function of time for a period of 150 minutes. A logarithmic oxidation growth with two different rate constants is obtained. X-ray diffraction analysis of 50 nm thick films at various temperatures indicated a solid solution of chromium and iron-chromium oxides. A decrease of lattice spacing observed with temperature increase is attributed to composition/structural changes near the metal-oxide interface. Microstructures of these film surfaces reveal the formation of bluish green crystals of Cr2O3 at 773 K.

Approximately one quarter to one third of patients with cancer are either not candidates for biopsies, or have insufficient tissue samples gathered during their initial biopsy. These factors may limit the ability to provide a required diagnosis at the molecular level and recommend EGFR TKI therapy. The goal of this study was to develop and validate rapid, robust and highly sensitive blood-based assays that identify the EGFR sensitizing mutations L858R and exon 19 deletion (E746 - A750) in circulating DNA isolated from the plasma of patients previously diagnosed with non-small cell lung cancer (NSCLC). Additional studies were conducted to similarly detect the EGFR resistance mutation, T790M which could have implications for 3rd generation EGFR TKI therapies. The developed test is comprised of three components: 1. a whole-blood specimen collection kit that ships at ambient temperature; 2. cell free DNA (cfDNA) isolated from patient plasma samples and processed using a droplet digital PCR workflow and analyses, and 3. LIMS software for report generation. The EGFR mutation status result (classification label) is reported within 72 hours of sample shipment. Components of the assay development for the EGFR sensitizing and resistance mutation tests included: (a) ambient temperature ship stability of the sample collection kit, (b) the test system performance verification/method development studies with pre-qualified DNA standards (n = 10) and human donor samples (n = 248) and (c) sensitivity and specificity analyses with tissue. The lower limit of detection of the test system was established at 0.02% minor allele frequency. The precision (inter-day, intra-day performance) was established with validated standards as well as with human donor samples (SD < 10% overall for all three assays). The robustness for all assays was evaluated over 20 consecutive days of testing, with 100% of tests passing (SD < 2.0%). Sensitivity for the EGFR sensitizing and resistance mutations was shown to be 95.7% and 86.7% respectively. Specificity of the assays for their targets was established at 100% with no cfDNA in any of the healthy donors, and no cases of known tissue negative cancer donors yielding positive results to date (n = 159). In conclusion, we have developed three highly sensitive blood-based assays to identify sensitizing and resistance mutations for EGFR (L858R and exon 19 deletion E746 - A750, and T790M, respectively) in circulating DNA in plasma from patients previously diagnosed with NSCLC. These results indicate that plasma cfDNA is a reliable source for EGFR mutation analysis in clinical practice, especially for those patients unable to provide tissue-based samples. The results can be delivered to physicians within 72 hours from sample shipment and may facilitate more immediate patient management. Citation Format: Trudi Foreman, Kristina Koch, Amanda Weaver, Nicholas Dupuis, Steven Arrivo, Ubaradka Sathyanarayana, Hestia Mellert, Gary Pestano. Development of a rapid blood-based test for EGFR sensitizing and resistance mutations in NSCLC. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B137.