A Randomized Controlled Trial to Compare Peripherally Inserted Central Catheter Tunnel Lengths in Adult Patients With Cancer.

BACKGROUND: Evidence is insufficient on the effect of tunnel lengths on tunneled peripherally inserted central catheter (PICC) placement in adult patients with cancer. OBJECTIVES: The primary objective was to explore whether there is an optimal PICC tunnel length to reduce the risk of PICC-related complications. The secondary objective was to compare patients' pain and comfort levels during catheter placement with different tunnel lengths. METHODS: Two hundred patients were randomly assigned to groups based on PICC tunnel length. Data collected included baseline characteristics, catheter-related characteristics, PICC-related complications, and patients' pain and comfort levels. FINDINGS: Patients with 4 cm, 5 cm, and 6 cm PICC tunnel lengths had a longer catheter dwell time and fewer PICC-related complications. No significant differences were found among all groups regarding patients' pain and comfort levels. The results suggest that a tunneled PICC is safe and effective. A tunnel length longer than 4 cm is recommended for tunneled PICC placement.

Exosomal miR-196a-5p enhances radioresistance in lung cancer cells by downregulating NFKBIA.

Radiation therapy is recognized as an effective modality in the treatment of lung cancer, but radioresistance resulting from prolonged treatment reduces the chances of recovery. MicroRNAs (miRNAs) play a pivotal role in radiotherapy immunity. In this study, we aimed to investigate the mechanism by which miR-196a-5p affects radioresistance in lung cancer. The radioresistant lung cancer cell line A549R26-1 was established by radiation treatment. Cancer-associated fibroblasts (CAFs) and normal fibroblasts (NFs) were observed by microscopy, and the expression levels of CAF-specific marker proteins were detected by immunofluorescence. The shape of the exosomes was observed by electron microscopy. A CCK-8 assay was used to detect cell viability, while clone formation assays were used to detect cell proliferative capacity. Flow cytometry was performed to investigate apoptosis. The binding of miR-196a-5p and NFKBIA was predicted and further verified by the dual luciferase reporter experiment. qRT-PCR and western blotting were used to detect gene mRNA and protein levels. We found that exosomes secreted by CAFs could enhance lung cancer cell radioresistance. Moreover, miR-196a-5p potentially bound to NFKBIA, promoting malignant phenotypes in radioresistant cells. Furthermore, exosomal miR-196a-5p derived from CAFs increased radiotherapy immunity in lung cancer. Exosomal miR-196a-5p derived from CAFs enhanced radioresistance in lung cancer cells by downregulating NFKBIA, providing a new potential target for the treatment of lung cancer.

Identifying the impact of the Zone Insertion MethodTM (ZIMTM): A randomized controlled trial.

BACKGROUND: In 2011, Dawson proposed the Zone Insertion MethodTM (ZIMTM) to identify the optimal peripherally inserted central catheters (PICCs) insertion site in the upper arm. However, data on the effectiveness and safety of the ZIMTM in guiding PICC placement in Chinese population is limited. METHODS: In this randomized controlled trial, 120 cancer patients were randomly assigned to the upper portion of the red zone (RZ), the green zone (GZ) and the lower portion of the yellow zone (YZ) groups (at a 1:1:1 ratio). The aim was to compare the degree of patient comfort and the incidence of major PICC complications among the three insertion zones based on the ZIMTM in a Chinese Cancer Center. (Clinical Trials. Gov number, ChiCTR1900024111). RESULTS: A total of 118 catheters were inserted in 118 patients (2 patients were lost to follow-up). After the 1-month follow-up, patients randomly assigned to the YZ group had a higher degree of comfort with a lower score than those assigned to the other two zone groups: 30.21±3.16 in the YZ group versus 31.65±2.51 in the RZ group and 31.59±2.92 in the GZ group (P=.046). The incidence of thrombosis (10/40, 25%) and occlusion (4/40, 10%) in the RZ, which were significantly higher than those in the other two zone groups (χ2 =7.368, P=.02; χ2 =5.778, P =.03), whereas the risk in the GZ group was similar to that in the YZ group. The incidence of contact dermatitis in the GZ group was significantly higher than that of the other two zone groups (χ2=12.873, P=.001). CONCLUSIONS: This study found that the lower portion of YZ seems to be another suitable PICC insertion site for a higher degree of comfort and a lower risk of occlusion and thrombosis, which broadens the choice of PICC insertion sites in the upper arm for clinical practice.

Application and effect evaluation of multidisciplinary team management model: On central venous access device associated skin impairment based on Delphi method.

OBJECTIVE: To establish a multidisciplinary management model based on Delphi method to guide nursing practice and reduce the incidence of CVAD-associated Skin Impairment (CASI) in tumor patients. METHODS: On the basis of literature review and focus group interview, the initial item pool of CASI management model for cancer patients was determined. The Delphi method was used to conduct two rounds of letter consultation with 36 authoritative and representative experts to determine the content and weight of indicators of CASI multidisciplinary management model for cancer patients. RESULTS: Most of the research group were experts with bachelor degree or above. More than 90% of experts have worked for more than 10 years; Areas of expertise include oncology care, venous therapy, wound stomatology, and dermatology. The recovery rate of the two rounds of expert correspondence questionnaire was 100%. The authority coefficient of experts was 0.898, indicating a good degree of authority. Kendall's harmony coefficients were 0.193 and 0.250, with statistically significant differences (p < 0.001). After two rounds of expert letter consultation, a multidisciplinary management model of CASI for cancer patients was initially formed, which included 15 first-level prevention indexes and 38 second-level prevention indexes of CASI for cancer patients. There were 9 first-level indexes and 16 second-level indexes of CASI treatment in tumor patients. CONCLUSION: Cancer patients based on Delphi method to construct CASI multidisciplinary management model has high reliability and scientificity, multidisciplinary management model in the management of patients with tumor CASI exploration will provide new methods for central venous catheter nursing and the new way of thinking, will also be intravenous fluids will provide a scientific basis for professional development and quality improvement and practical experience.

Identification of the Immune Cell Infiltration Landscape in Head and Neck Squamous Cell Carcinoma (HNSC) for the Exploration of Immunotherapy and Prognosis.

It is generally believed that the majority of head and neck cancers develop in the mucosal epithelial cells of the mouth, pharynx, and larynx, which is collectively known as head and neck squamous cell carcinoma (HNSC). As a complex pathological process, HNSC develops through a variety of cellular and molecular events. Cancerous cells and immune cells infiltrating tumors are the main components of the tumor microenvironment. However, infiltration of HNSCs by the immune system has not been determined to date. In this work, we proposed computational algorithms to identify different immune subtypes. An analysis of the Cancer Genome Atlas (TCGA) database revealed gene expression profiles and corresponding clinical information. In HNSC patients, two immune-related genes (ZAP70 and IGKV2D-40) may be targets for immunotherapy, and these genes appear to be closely related to the prognosis. Several immunological subtypes were associated with immune function, immune checkpoints, and prognostic factors in HNSCs. Furthermore, ZAP70 is closely related to the overall survival (OS), progress-free interval (PFI), and disease-specific survival (DSS) of HNSC patients. The potential pathways that are associated with ZAP70 were found to have included adaptive immune response, response to oxidative stress, DNA replication, and lipid binding. This study provides a theoretical foundation for developing immunotherapy drugs for HNSC patients. By evaluating larger cohorts, we can gain a deeper understanding of immunotherapy and provide direction for current research on immunotherapy strategies in HNSCs.

Effect of video-assisted education on informed consent and patient education for peripherally inserted central catheters: a randomized controlled trial.

OBJECTIVE: To evaluate the effects of a video-assisted education intervention on informed consent and patient education for peripherally inserted central catheters (PICCs). METHODS: We conducted a randomized controlled trial comparing the effects on informed consent of video-assisted patient education and traditional face-to-face discussion in a catheter outpatient ward of a cancer centre in Guangzhou, China, in 2018. Participants were 140 patients randomly allocated (1:1 ratio) to two groups: video-assisted or traditional intervention. General information, patient retention of PICC-related information, working time spent by nurses on the procedure, and patient and nurse satisfaction with the procedure were assessed. RESULTS: The time used for informed consent was significantly shorter in the experimental group (1.02 ± 0.24 minutes) than in the control group (6.87 ± 1.10 minutes). The time used for PICC-related education was significantly shorter in the experimental group (1.03 ± 0.28 minutes) than in the control group (5.11 ± 0.57 minutes). Nurses' degree of satisfaction with the procedure was significantly higher in the experimental group (4.10 ± 0.57) than in the control group (2.60 ± 0.70). CONCLUSION: The use of video-assisted informed consent and patient education in this cancer centre decreased nurses' working time and improved nurses' satisfaction.Clinical trial registration number: ChiCTR1800015664.

Serum exosomal miR-7977 as a novel biomarker for lung adenocarcinoma.

Exosomal microRNAs (miRNAs) have great potentials as a novel biomarker to predict lung cancer. We applied a miRNA microarray to identify aberrantly expressed serum exosomal miRNAs as candidate biomarkers for patients with lung adenocarcinoma (LUAD). Compared with the normal control, 31 exosomal miRNAs were found to be upregulated and 29 exosomal miRNAs were downregulated in the serum of LUAD respectively. Then, 10 dysregulated exosomal miRNAs expression levels in serum were further validated via qRT-polymerase chain reaction. Notably, exosomal miR-7977 was highest expressed and miR-98-3p was lowest expressed in the patients with LUAD, and exosomal miR-7977 showed significant correlation with the N stage and TNM stage with patients with LUAD (P < .05). Receiver operating characteristic curve showed that the abundant level of exosomal miR-7977 may predict LUAD with an area of under the curve (AUC) of 0.787. In comparison with exosomal miR-7977, exosomal miR-98-3p had a smaller area (0.719). The combination of exosomal miR-7977 and miR-98-3p improved the AUC to 0.816. Furthermore, in vitro experiments revealed that inhibition of miR-7977 enhanced the proliferation, invasion, and inhibited apoptosis in A549 cells, the opposite results were performed by miR-7977 mimics. In conclusion, exosomal miR-7977 was identified as a novel biomarker for patients with LUAD and may play as a tumor suppressor in lung cancer.

Circular RNAs expression profiles in plasma exosomes from early-stage lung adenocarcinoma and the potential biomarkers.

This study aimed to identify differential circular RNA (circRNA) in the plasma exosomes of patients with lung adenocarcinoma (LUAD) using high-throughput sequencing. First, exosomes were isolated using an exosome isolation kit and confirmed by Western blotting, transmission electron microscopy, and NanoSight Assay. Subsequently, plasma circRNA expression profiles were screened by high-throughput sequencing and confirmed by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR) and Sanger sequencing. Finally, the circRNA-miRNA-mRNA network was performed to forecast the potential function of circRNAs. The result of high-throughput sequencing data documented that 182 differentially expressed exosomal circRNAs in all were screened, which included 105 that were upregulated and 78 that were downregulated in LUAD patients plasma compared with controls. The four upregulated circRNAs including circ_0001492, circ_0001346, circ_0000690, and circ_0001439 were identical to the sequencing data by qRT-PCR, and their latent circRNA-miRNA-mRNA interactions were exhibited. Taken together, our study firstly revealed the altered exosomal circRNA expression from plasma samples in patients with LUAD and supports the need for exploring their potential as biomarkers and the pathological effects of lung cancer.

[Human THP-1 macrophages activated by exosomes derived from lung adenocarcinoma cells promote lung cancer cell invasion].

Objective To examine the effect of exosomes derived from lung adenocarcinoma cells on macrophage polarization and the change of cytobiological behaviors in lung cancer cells induced by activated macrophages. Methods Exosomes derived from lung adenocarcinoma cells were extracted by exosomes extraction kit. The expression of exosomal markers including CD9 and CD63 was detected by Western blot analysis. After THP-1 cells were treated with 100 ng/mL phorbol ester (PMA) for 48 hours, the macrophage marker of CD68 was detected by real-time quantitative PCR (RT-qPCR). Following 24-hour treatment of macrophages with the exosomes (200 µg/mL), the mRNA levels of transforming growth factor ß (TGF-ß), tumor necrosis factor α (TNF-α), inducible nitric oxide synthase (iNOS) and CD163 were detected by RT-qPCR, and the protein levels of IL-6, IL-8 and IL-10 were measured by IMMULITE 1000. The macrophages after exosome treatment were co-cultured with A549 or H1299 cells. The invasion of lung adenocarcinoma cells was tested by TranswellTM assay and the mRNA levels of MMP9, MMP2 in lung adenocarcinoma cells were detected by RT-qPCR. Results CD9 and CD63 were highly expressed in exosomes. The THP-1 cells after PMA induction produced a high level of CD68. After the macrophages were treated with exosomes, the expression of iNOS decreased and the expression of CD163, TNF-α, IL-6, IL-8 and IL-10 significantly increased in the macrophages. The co-culture of macrophages with A549 and H1299 after exosome treatment enhanced significantly the invasion ability of lung adenocarcinoma cells and increased the levels of MMP2 and MMP9. Conclusion The exosomes derived from lung adenocarcinoma cells can activate macrophages to exhibit a mixed M1/M2 phenotype, thus promot the invasion of lung cancer cells.

A comprehensive risk assessment of human inhalation exposure to atmospheric halogenated flame retardants and organophosphate esters in an urban zone.

Inhalation exposure to flame retardants used as additives to minimize fire risk and plasticizers is ubiquitous in human daily activities, but has not been adequately assessed. To address this research gap, the present study conducted an assessment of human health risk for four age groups through inhalation exposure to size fractionated particle-bound and gaseous halogenated flame retardants (polybrominated diphenyl ethers (PBDEs) and alternative halogenated flame retardants (AHFRs)) and organophosphate esters (OPEs) at indoor and outdoor environments (school, office, and residence) in three districts of a megacity (Guangzhou, China). Results demonstrated that OPEs were the dominant components among all targets. Indoor daily intakes of PBDEs and OPEs were 13-16 times greater than outdoor levels for all age groups. Gaseous OPEs contributed significantly greater than particle-bound compounds to daily intakes of all target compounds. Based on the different life scenarios, hazard quotient (HQ) and incremental life cancer risk (ILCR) from adults exposure to PBDEs and OPEs in indoor and outdoor settings were the greatest, followed by adolescents, children, and seniors. The estimated HQ and ILCR for all age groups both indoors and outdoors were lower than the safe level (HQ = 1 and ILCR = 10-6), indicating that the potential health risk for local residents in Guangzhou via inhalation exposure to atmospheric halogenated flame retardants and OPEs was low.

Prognostic roles of mRNA expression of peroxiredoxins in lung cancer.

BACKGROUND: The peroxiredoxin (PRDX) protein family is involved in cancer cell invasion and metastasis, but its prognostic value in lung cancer remain elusive. METHODS: In this report, we accessed the overall survival (OS) of each individual PRDX mRNA expression through the Kaplan-Meier plotter (KM plotter) database, in which updated gene expression data and survival information include a total of 1,926 lung cancer patients. RESULTS: Our results indicated that PRDX1 and PRDX2 mRNA expressions were associated with improved OS in all lung cancer patients especially in lung adenocarcinoma patients, whereas PRDX5 and PRDX6 mRNA expressions were associated with poor OS in all lung cancer patients. In addition, the prognostic value of PRDXs in the different clinicopathological features according to smoking status, pathological grades, clinical stages, and chemotherapeutic treatment of lung cancer patients was further assessed in the KM plotter database by the multivariate cox regression analysis. CONCLUSION: Our finding will elucidate the prognostic role of PRDXs in lung cancer and might promote development of PRDX-targeted inhibitors for the treatment of lung cancer.

Challenging pharmaceutical analyses by gas chromatography with vacuum ultraviolet detection.

Vacuum ultraviolet (VUV) detector for gas chromatography (GC) provides qualitative spectral information from 125 nm to 240 nm. In this article, this information was applied to facilitate the development of a GC method for challenging pharmaceutical applications. Seven organic solvents were screened for trace level water content using VUV detection at 168 nm, and the results were used to identify n-hexane as a suitable diluent for 4-ethoxy-1,1,1-trifluoro-3-buten-2-one (ETFBO), a water reactive compound. Selective detection of compounds of interest was demonstrated by varying detection wavelengths. All compounds were detected at 145 nm except for one unknown impurity, which co-eluted with n-hexane solvent. This impurity was detected at 225 nm, where n-hexane has no absorbance. In addition, the VUV spectra were used to: 1) accurately track peaks during early method development; 2) detect co-eluting peaks; 3) match peak identity in a sample vs. a standard; and 4) assess peak purity. With the universal detectability, qualitative spectral information and ease of use, VUV will become a versatile tool for GC for both method development and routine analysis.

Assessment of airborne polycyclic aromatic hydrocarbons in a megacity of South China: Spatiotemporal variability, indoor-outdoor interplay and potential human health risk.

Although a number of studies have assessed the occurrence of atmospheric polycyclic aromatic hydrocarbons (PAHs) in indoor environment, few studies have systemically examined the indoor-outdoor interplay of size-dependent particulate PAHs and potential health risk based on daily lifestyles. In the present study, size-dependent particle and gaseous samples were collected both indoors and outdoors within selected schools, offices and residences located in three districts of Guangzhou, China with different urbanization levels during the dry and wet weather seasons. Results from measurements of PAHs showed that higher total PAH concentrations occurred in residential areas than in other settings and in indoor than in outdoor environments. Compositional profiles and size distribution patterns of particle-bound PAHs were similar indoors and outdoors, predominated by 4-and 5-ring PAHs and the 0.56-1.0 µm particle fraction. Statistical analyses indicated that outdoor sources may have contributed to 38-99% and 62-100% of the variations for indoor particle-bound and gaseous PAH concentrations, respectively. Incremental life cancer risk (ILCR) from human exposure to indoor and outdoor PAHs based on different lifestyles followed the order of adults > children > adolescents > seniors. All average ILCR values for four age groups were below the lower limit of the Safe Acceptable Range (10-6). In addition, the ILCR value for adults (average: 7.2 × 10-7; 95% CI: 5.4 × 10-8‒2.5 × 10-6), estimated from outdoor air PAH levels with 24-h exposure time, was significantly higher than our assessment results (average: 5.9 × 10-7; 95% CI: 6.3 × 10-8‒1.9 × 10-6), suggesting the significance of assessing human inhalation exposure risks of indoor and outdoor PAHs in urban air based on daily lifestyles.

Liver Microvascular Injury and Thrombocytopenia of Antibody-Calicheamicin Conjugates in Cynomolgus Monkeys-Mechanism and Monitoring.

Purpose: Adverse reactions reported in patients treated with antibody-calicheamicin conjugates such as gemtuzumab ozogamicin (Mylotarg) and inotuzumab ozogamicin include thrombocytopenia and sinusoidal obstruction syndrome (SOS). The objective of this experimental work was to investigate the mechanism for thrombocytopenia, characterize the liver injury, and identify potential safety biomarkers.Experimental Design: Cynomolgus monkeys were dosed intravenously at 6 mg/m2/dose once every 3 weeks with a nonbinding antibody-calicheamicin conjugate (PF-0259) containing the same linker-payload as gemtuzumab ozogamicin and inotuzumab ozogamicin. Monkeys were necropsied 48 hours after the first administration (day 3) or 3 weeks after the third administration (day 63).Results: PF-0259 induced acute thrombocytopenia (up to 86% platelet reduction) with nadirs on days 3 to 4. There was no indication of effects on megakaryocytes in bone marrow or activation of platelets in peripheral blood. Microscopic evaluation of liver from animals necropsied on day 3 demonstrated midzonal degeneration and loss of sinusoidal endothelial cells (SECs) associated with marked platelet accumulation in sinusoids. Liver histopathology on day 63 showed variable endothelial recovery and progression to a combination of sinusoidal capillarization and sinusoidal dilation/hepatocellular atrophy, consistent with early SOS. Among biomarkers evaluated, there were early and sustained increases in serum hyaluronic acid (HA) that correlated well with serum aspartate aminotransferase and liver microscopic changes, suggesting that HA may be a sensitive diagnostic marker of the liver microvascular injury.Conclusions: These data support the conclusion that target-independent damage to liver SECs may be responsible for acute thrombocytopenia (through platelet sequestration in liver sinusoids) and development of SOS. Clin Cancer Res; 23(7); 1760-70. ©2016 AACR.

Potential health risk for residents around a typical e-waste recycling zone via inhalation of size-fractionated particle-bound heavy metals.

Health risk of residents dwelling around e-waste recycling zones has been a global concern, but has not been adequately examined. The present study was intended to evaluate the potential health risk of residents through inhalation exposure to size-fractionated particle-bound heavy metals in a typical e-waste recycling zone, South China. Anthropogenic metals (Zn, Se, Pb, Sb, As, and Cd) were predominantly enriched in fine particles (Dp<1.8µm), whereas the crustal elements (Ti, Fe, and Co) tended to accumulate in coarse particles (Dp>1.8µm). Although the daily inhalation intakes of the target metals were significantly lower than those through food consumption and ingestion of house dust, the hazard quotients of total metals for adults (95% CI: 1.0-5.5) and children (95% CI: 3.0-17) were greater than 1. Moreover, the incremental lifetime cancer risks of five carcinogenic metals (Cr, Co, Ni, As, and Cd) for adults and children were 1.3×10(-3) (95% CI: 4.1×10(-4)-3.0×10(-3)) and 3.9×10(-3) (95% CI: 1.3×10(-3)-8.6×10(-3)), respectively, substantially higher than the acceptable cancer risk range of 10(-6)-10(-4). All these findings suggested that health risks were high for local residents dwelling around the e-waste recycling zone through inhalation exposure to particle-bound heavy metals, for both adults and children.

Isolation and characterization of three TaYUC10genes from wheat.

YUCCA protein participates in a key rate-limiting step in the tryptophan-dependent pathway for auxin biosynthesis and is involved in numerous processes during plant development. In this study, the genomic and cDNA sequences of three TaYUC10 homoeologous genes were isolated. These sequences showed a very high conservation in coding region and the exon/intron structure, whereas their intron lengths were different. The cDNA and polypeptide chains of the three TaYUC10 genes were highly similar. These genes were most homologous to BdYUC10. Location analysis showed that TaYUC10.1 was present in chromosome 5BL. TaYUC10.3 was expressed in all parts of the wheat, but was predominant in the reproductive organs of mature wheat, such as flowering spikelets or fertilized embryos. In the fertilized embryos 28d post-anthesis, expression of TaYUC10.3 was clearly increased with the development of seeds. This indicates that TaYUC genes may play a vital role in seed development. TaYUC10.3 overexpressed in Arabidopsis had a typical phenotype, excessive auxin accumulation also seen in higher plants, and showed increased spacing of silique and downward curling of the blade margin. Sterility was observed in adult transgenic plants, becoming more severe in late development. The floral structures of sterile plants were not integrated. TaYUC10 may be required for numerous wheat growth processes, including flower and seed development.

Effects of age on noninvasive assessments of vascular function in nonhuman primates: implications for translational drug discovery.

BACKGROUND: Endothelium-dependent flow mediated dilation (FMD) and pulse-wave velocity (PWV), are used as measures of vascular health and predictors of cardiovascular risk in clinical studies, and both are age-dependent. Numbers of circulating endothelial microparticles (EMPs) and endothelial progenitor cells (EPCs) are also associated with cardiovascular risk, but independent of age in humans. The use of these measurements for pre-clinical assessment of drug cardiovascular safety and efficacy in non-human primates (NHPs) may promote the translation of drug-induced effects on vascular function to clinic outcomes. However, in NHPs, the age effects on the non-invasive measurements of FMD and PWV and the relationship of EMPs/EPCs with FMD are unknown. METHODS: A non-invasive, clinically-relevant approach to assess FMD and PWV was used to examine their relationship with age and with EMPs/EPCs in NHPs. The effects on FMD of nicotine and rosiglitazone were evaluated in senescent primates in an effort to validate our FMD method for pre-clinical assessment of vascular function. RESULTS: FMD and PWV methods were established in a colony (n = 25) of metabolically healthy, cynomolgus monkeys ranging in age from 6 to 26 years. FMD, defined as the percent change, at 1 min of cuff release, from baseline vascular diameter (0.15 ± 0.03 cm), had a strong, negative correlation with age (r = -0.892, p < 0.0001), ranging from 6% to 33%. PWV positively correlated with age (r = 0.622, p < 0.002) in the same healthy monkeys. Nicotine and rosiglitazone, were evaluated in subsets of senescent primates (mean age 16.3 ± 1.5[SEM] years). Rosiglitazone significantly improved FMD (21.0 ± 1.6% vs. vehicle 16.3 ± 1.6%, p < 0.01) without changing baseline diameters, and coincided with a significant increase in circulating numbers of endothelial progenitor cells (CD45-CD31 + CD34 + VEGFR2+ 7.1 ± 1.3 vs. 4.8 ± 1.1 counts/µl) and a decrease in endothelial microparticles (CD45-CD42a-CD54+ 26.7 ± 11.1 vs. 62.2 ± 9.8 counts/µl)(p < 0.05). Conversely, FMD was significantly reduced with nicotine (8.7 ± 1.4% vs. vehicle 20.1 ± 2.2%, p < 0.05). CONCLUSIONS: Adult NHPs demonstrate the characteristic linear relationship between age and vascular function using the non-invasive clinically-related measurements of FMD and PWV. However, numbers of circulating EMPs and EPCs did not correlate with age. Endothelial function assessed with FMD, together with EMPs/EPCs assessment, may serve as a novel approach for translational research and therapeutic discovery. Age should be considered in the study design or data analyses when FMD or PWV is used in NHPs.

Targeting of 4-1BB by monoclonal antibody PF-05082566 enhances T-cell function and promotes anti-tumor activity.

4-1BB (CD137, TNFRSF9) is a costimulatory receptor expressed on several subsets of activated immune cells. Numerous studies of mouse and human T cells indicate that 4-1BB promotes cellular proliferation, survival, and cytokine production. 4-1BB agonist mAbs have demonstrated efficacy in prophylactic and therapeutic settings in both monotherapy and combination therapy tumor models and have established durable anti-tumor protective T-cell memory responses. PF-05082566 is a fully human IgG2 that binds to the extracellular domain of human 4-1BB with high affinity and specificity. In preclinical studies, this agonist antibody demonstrated its ability to activate NF-κB and induce downstream cytokine production, promote leukocyte proliferation, and inhibit tumor growth in a human PBMC xenograft tumor model. The mechanism of action and robust anti-tumor efficacy of PF-05082566 support its clinical development for the treatment of a broad spectrum of human malignancies.

The Melanoma Vascular Mimicry Phenotype Defined in Gene Expression and Microsome Sequencing Analysis.

The phenomenon of vasculogenic mimicry in melanoma has been recently described to be an important factor relating to melanoma progression. Large scale gene expression profiling by real-time quantitative RT-QPCR of a panel of 40 normal tissues and 54 cancer cell lines revealed that two genetically related melanoma cell lines, one derived from a primary lesion Hs.688(A) and one derived from a lymph node metastasis Hs.688(B), displayed a unique expression pattern when compared to other cancer cell lines and tissue samples in the panel. Quantitative-RT-PCR data indicated that these melanoma cells expressed a number of activated endothelial cell-associated genes such as tissue inhibitors of matrix metalloproteinases TIMP-2, matrix metalloproteinase (MMP-1, MMP-2), thrombospondin 1 (TSP1), proto-oncogene c-MET and vascular endothelial growth factor (VEGF). To examine the gene expression profile of these unique melanoma cells in greater depth, cDNA libraries were made from isolated microsome complexes to enrich those transcripts that were destined to be translated into cell surface or secreted proteins. High throughput sequencing analysis revealed that this library contained over 7000 cDNAs and was enriched by over 80% of secreted or membrane-bound proteins. The presence in the cDNA library of genes such as acetyl LDL receptor, tumor endothelial markers-1, 5 and 8 (TEMs), flow-induced endothelial G protein coupled receptor-1 and VEGF-related protein (VRP), all of which are known to be expressed uniquely by endothelial cells, supported the hypothesis that Hs.688(A) and Hs.688(B) cells were mimicking an activated vascular phenotype. Ultimately the goal is to investigate the biological roles of endothelial cell-associated genes in the behavior of Hs.688(A) and Hs.688 (B) melanoma cells.

Simultaneous gene expression analysis of steady-state and actively translated mRNA populations from osteosarcoma MG-63 cells in response to IL-1alpha via an open expression analysis platform.

Pro-inflammatory cytokines play a key role in various forms of metabolic bone diseases, including osteopenia and osteoporosis. Human MG-63 cells treated with IL-1alpha were used as a model system to identify potential marker genes that are differentially expressed. This study is designed to quantitate gene expression of actively translated mRNAs as compared to the steady-state mRNA population. Both steady-state mRNAs and actively translated mRNAs from control MG-63 cells and MG-63 cells treated with IL-1alpha were isolated and converted to cDNA. The gene expression analysis from these samples was then quantitated with an open expression analysis platform with no requirement for a priori knowledge of sequence information. As a result, many differentially regulated genes were discovered via IL-1alpha treatment. Some of the genes have been described previously as playing important roles in the regulation of inflammation and cell adhesion. These comparisons provided a panoramic overview of gene expression at both the total transcript and post-transcriptional levels. In addition, the quantitation of actively translated mRNAs associated with polysomes also provided a better estimation of protein expression levels. This methodology allows for the identification of genes acutely regulated during translation. Furthermore, the process may aid in the identification of new drug targets or biomarkers.