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In this study, the optimized niosomal formulation containing paclitaxel using non-ionic surfactants and cholesterol was designed and its cytotoxic effects against different breast cancer cell lines and apoptosis gene expression analysis were also investigated.Due to enhancing equation variables, the Box-Behnken method has been applied. Lipid/drug molar ratio, the amounts of Span 60, and cholesterol were selected as the target for optimization. The particle size of niosome loaded paclitaxel and entrapment efficiency proportion have been considered in the role of dependent variables. Then the cytotoxic activity of the optimized formulation was evaluated using an MTT assay against different breast cancer cell lines including MCF-7, T-47D, SkBr3, and MDA-MB-231. The expression level of Bax and Bcl-2 apoptosis genes was determined by Real-Time PCR. In this study, the optimized niosomal formulation revealed that the synthesized niosomes had a spherical appearance and had an average size of 192.73 ± 5.50 nm so that the percentage of drug loading was 94.71 ± 1.56%. Moreover, this formulation showed a controlled and slowed release of paclitaxel at different pH (7.4, 6.5, and 5.4). The cytotoxicity results demonstrated that cell viability in all concentrations of niosome loaded paclitaxel had profound cytotoxic effects on all studied breast cancer cell lines compared to the free paclitaxel (p 0.05). In addition, the expression of apoptosis genes was much higher than that of free paclitaxel indicating the susceptibility of cells to apoptosis.As a result, niosomal formulations containing paclitaxel can be used as a new drug delivery system to increase cytotoxicity and treatment of breast cancer in the upcoming future.

The limbal stem cells niche (LSCN) is an optimal microenvironment that provides the limbal epithelial stem cells (LESCs) and strictly regulates their proliferation and differentiation. Disturbing the LSCN homeostasis can lead to limbal stem cell dysfunction (LSCD) and subsequent ocular surface aberrations, such as corneal stromal inflammation, persistent epithelial defects, corneal neovascularisation, lymphangiogenesis, corneal opacification, and conjunctivalization. As ocular surface disorders are considered the second main cause of blindness, it becomes crucial to explore different therapeutic strategies for restoring the functions of the LSCN. A major limitation of corneal transplantation is the current shortage of donor tissue to meet the requirements worldwide. In this context, it becomes mandatory to find an alternative regenerative medicine, such as using cultured limbal epithelial/stromal stem cells, inducing the production of corneal like cells by using other sources of stem cells, and using tissue engineering methods aiming to produce the three-dimensional (3D) printed cornea. Limbal epithelial stem cells have been considered the magic potion for eye treatment. Epithelial and stromal stem cells in the limbal niche hold the responsibility of replenishing the corneal epithelium. These stem cells are being used for transplantation to maintain corneal epithelial integrity and ultimately sustain optimal vision. In this review, we summarised the characteristics of the LSCN and their current and future roles in restoring corneal homeostasis in eyes with LSCD.

Adeno-associated virus (AAV)-based gene therapies, exemplified by the approved therapy for spinal muscular atrophy, have the potential to deliver disease-course-altering treatments for central nervous system (CNS) indications. However, several clinical trials have reported severe adverse events including patient deaths following high-dose systemic administration for muscle-directed gene transfer, highlighting the need to explore approaches utilizing lower doses when targeting the CNS. Animal models of disease provide insight into the response to new AAV therapies. However, translation from small to larger animals and eventually to humans is hampered by anatomical and biological differences across the species and their impact on AAV delivery. We performed a literature review of preclinical studies of AAV gene therapy biodistribution following cerebrospinal fluid (CSF) delivery (intracerebroventricular, intra-cisterna magna, and intrathecal lumbar). The reviewed literature varies greatly in the reported biodistribution of AAV following administration into the CSF. Differences between studies including animal model, vector serotype used, method used to assess biodistribution, and route of administration, among other variables, contribute to differing outcomes and difficulties in translating these preclinical results. For example, only half of the published AAV-based gene therapy studies report vector copy number, the most direct readout following administration of a vector; none of these studies reported details such as the empty:full capsid ratio and quality of encapsidated genome. Analysis of the last decade's literature focusing on AAV-based gene therapies targeting the CNS underscores limitations of the body of knowledge and room for continued research. In particular, there is a need to understand the biodistribution achieved by different CSF-directed routes of administration and determining if specific cell types/structures of interest will be transduced. Our findings point to a clear need for a more systematic approach across the field to align the assessments and elements reported in preclinical research to enable more reliable translation across animal models and into human studies.

Adeno-associated viruses (AAVs) are being increasingly used as gene therapy vectors in clinical studies especially targeting central nervous system (CNS) disorders. Correspondingly, host immune responses to the AAV capsid or the transgene-encoded protein have been observed in various clinical and preclinical studies. Such immune responses may adversely impact patients' health, prevent viral transduction, prevent repeated dosing strategies, eliminate transduced cells, and pose a significant barrier to the potential effectiveness of AAV gene therapy. Consequently, multiple immunomodulatory strategies have been used in attempts to limit immune-mediated responses to the vector, enable readministration of AAV gene therapy, prevent end-organ toxicity, and increase the duration of transgene-encoded protein expression. Herein we review the innate and adaptive immune responses that may occur during CNS-targeted AAV gene therapy as well as host- and treatment-specific factors that could impact the immune response. We also summarize the available preclinical and clinical data on immune responses specifically to CNS-targeted AAV gene therapy and discuss potential strategies for incorporating prophylactic immunosuppression regimens to circumvent adverse immune responses.

Limbus-derived stromal/mesenchymal stem cells (LMSCs) are vital for corneal homeostasis and wound healing. However, despite multiple pre-clinical and clinical studies reporting the potency of LMSCs in avoiding inflammation and scarring during corneal wound healing, the molecular basis for the ability of LMSCs remains unknown. This study aimed to uncover the factors and pathways involved in LMSC-mediated corneal wound healing by employing RNA-Sequencing (RNA-Seq) in human LMSCs for the first time. We characterized the cultured LMSCs at the stages of initiation (LMSC−P0) and pure population (LMSC−P3) and subjected them to RNA-Seq to identify the differentially expressed genes (DEGs) in comparison to native limbus and cornea, and scleral tissues. Of the 28,000 genes detected, 7800 DEGs were subjected to pathway-specific enrichment Gene Ontology (GO) analysis. These DEGs were involved in Wnt, TGF-β signaling pathways, and 16 other biological processes, including apoptosis, cell motility, tissue remodeling, and stem cell maintenance, etc. Two hundred fifty-four genes were related to wound healing pathways. COL5A1 (11.81 ± 0.48) and TIMP1 (20.44 ± 0.94) genes were exclusively up-regulated in LMSC−P3. Our findings provide new insights involved in LMSC-mediated corneal wound healing.

Epileptic encephalopathies may arise from single gene variants. In recent years, next-generation sequencing technologies have enabled an explosion of gene identification in monogenic epilepsies. One such example is the epileptic encephalopathy SLC13A5 deficiency disorder, which is caused by loss of function pathogenic variants to the gene SLC13A5 that results in deficiency of the sodium/citrate cotransporter. Patients typically experience seizure onset within the first week of life and have developmental delay and intellectual disability. Current antiseizure medications may reduce seizure frequency, yet more targeted treatments are needed to address the epileptic and non-epileptic features of SLC13A5 deficiency disorder. Gene therapy may offer hope to these patients and better clinical outcomes than current available treatments. Here, we discuss SLC13A5 genetics, natural history, available treatments, potential outcomes and assessments, and considerations for translational medical research for an AAV9-based gene replacement therapy.

Introduction Although the clinical manifestations of severe haemophilia A (HA) are well studied, the challenges, if any, of living with mild HA are not clearly delineated to date. Aim To assess available evidence of clinical risks and societal/economic impacts of disease in adult patients with mild HA using a systematic literature review. Methods Prespecified study selection criteria were applied in a comprehensive literature search. Included studies varied in design and reported outcomes of interest for adults (≥13 years of age) with mild HA. Results Seventeen studies with a total of 3213 patients met eligibility criteria (published or presented in English, 1966‐2017). Most studies were observational, and the outcomes reported were too sparse and dissimilar to support a formal meta‐analysis. Mean annual bleeding rates ranged from 0.44 to 4.5 episodes per patient per year. Quality of life (QoL; SF‐36 General Health) was impacted compared to healthy controls. Health care costs and productivity were seldom assessed and no robust comparisons to healthy controls were available. Conclusion Quantifying outcomes for adult patients with mild HA remains challenging, with estimates of key QoL and cost data often based on small data sets and without comparison to population norms. Therefore, the clinical impact of mild haemophilia may be under‐represented and unmet needs may remain unaddressed. As paradigm‐changing therapies for HA emerge, stronger knowledge of mild HA can guide the development of care options that minimize burden and enhance the QoL for this segment of the haemophilia community, and for the haemophilia community in totality.

Corneal pathologies are a major cause of blindness and visual impairment, especially in the developing world. However, not only is there a global shortage of donor corneal tissue, but a significant proportion of these blinding pathologies also carry an unfavourable long-term prognosis for conventional corneal transplantation. In the last few decades, there has been a spurt of research on developing alternate approaches to address corneal blindness, including stem cell therapy. After the discovery of epithelial stem cells at the limbus, successful cell-based approaches to treat severe ocular surface disease were developed and have subsequently become widely practised across the world. More recently, mesenchymal stem cells were identified near the epithelial stem cells at the limbus, providing a unique opportunity to develop regenerative therapies for both corneal epithelial and stromal pathologies. This review firstly emphasises on qualifying limbal stem cells as either epithelial or mesenchymal and then summarises all the existing knowledge on both cell types and their individual roles in corneal regeneration. The review describes the history, indications, techniques, and outcomes of the different methods of limbal epithelial stem cell transplantation and elaborates on the potential applications of limbal mesenchymal stem cell therapy.

Introduction: The transparency of human cornea is critical for normal vision. It is maintained by a small population of stem cells present in the corneal limbus. Corneal diseases are a major cause of global blindness, second only to cataract in overall importance. They are usually treated by medications and surgical modalities that sometimes include expensive and potentially toxic medications with long-term side effects, particularly to prevent rejection of the corneal allograft. Therefore, cell based therapies may provide a promising alternative to overcome these limitations for the treatment of corneal diseases.Areas covered: This review article covers major sources, properties, and applications of mesenchymal stem cells in corneal reconstruction. This also includes a brief description of the structure, function, and immune privilege of the cornea; corneal stem cells, corneal diseases, available treatments, and potential of MSC-based cell therapy for corneal diseases.Expert opinion: Technical ad...

Several key attributes of a 3D integrated chip structure are analyzed in this work. Critical features related to the effect of the size of the substrate, heat sink, device layer, through silicon vias (TSVs), thermal interface material (TIM), and the pitch and arrangement of core processors and TSVs as well as variation of thermal conductivity and total heat dissipation and distribution of power within the device layers core processors are investigated in depth. The effects of variation of pertinent features of the 3D integrated circuit (IC) structure on thermal hotspots are established and an optimization route for its reduction is clarified. In addition, a revealing analysis that shows the effect of the number of layers in the 3D structure is presented. Furthermore, the features that have insufficient effect on reduction of thermal hotspots are also established and discussed.

A critical investigation is presented in this work to study the effect of nanoparticle addition, temperature, and nanoparticle size-dependence on the specific heat capacity of both conventional and molten salt-based nanofluids. The effects of temperature and nanoparticle volume fraction on the specific heat capacity of conventional nanofluids are in agreement in all studies cited in this review. Different correlations based on the available data were developed as a function of temperature and volume concentration only. However, the effect of nanoparticle size-dependence was ignored in these correlations. A general correlation for Al2O3–water nanofluids, one of the most commonly studied nanofluids, that takes into account the effect of temperature, volume fraction, and nanoparticle size-dependence was developed and verified in this review. Disagreement was reported for the results of the specific heat capacity of molten salt-based nanofluids. A number of studies showed an enhancement in the specific heat capacity of nanofluids using 1% concentration of nanoparticles by weight only. However, other studies have shown deterioration in the specific heat capacity of nanofluids compared with the base mixture using various volume concentrations of nanoparticles. Moreover, very few studies have demonstrated the effect of nanoparticle size-dependence on the specific heat capacity of molten salt nanofluids and disagreement in the results was reported in these studies. Few models based on the conventional specific heat model were developed to determine the specific heat capacity of molten salt nanofluids. These models suffer from the lack of knowledge of many terms in these equations which make them impractical. Different mechanisms were assumed in the literature to explain the abnormal behavior of molten salt nanofluids. Additional theoretical and experimental research studies are required to clarify the mechanisms responsible for specific heat capacity enhancement or deterioration in nanofluids.

The phenomenon of rarefaction in a micro-annulus filled with a porous medium is analyzed in the slip-flow regime. A local thermal non-equilibrium (LTNE) model is utilized to represent the energy transport within the porous medium. Exact solutions are derived for both the fluid and solid temperature distributions within the annulus. Two distinct cases of the thermal boundary conditions are considered, namely a constant heat flux at the outer wall and adiabatic inner wall (Case I) and vice versa (Case II). By eliminating the temperature difference between the fluid and solid phases, the local thermal equilibrium (LTE) model is theoretically proved to be a special case of the LTNE counterpart. Analytical predictions indicate that although the rarefaction leads to a reduction in the heat transfer, the effects of other thermophysical parameters such as the Biot number, the effective thermal conductivity ratio, the porous media shape factor and the annulus aspect ratio also play an important role. The results suggest that the configuration of Case II is superior to that of Case I from the heat transfer point of view.

An investigation is presented of free convection in horizontal concentric annuli of varying with inner shape where the inner and outer surfaces are kept at a constant temperature. The simulation is categorized into four groups based on the shape of the inner entity, which is either cylindrical, elliptical, square, or triangular. Flow and thermal fields are exhibited by means of streamlines and isotherms. Overall heat transfer correlations incorporating thermal radiation are established and presented in terms of the Nusselt number. It is observed that the surface radiation and presence of corners and larger top space enhances the heat transfer rate. As the reference temperature increases, surface radiation plays a more prominent role in the overall heat transfer performance.

Effects of hyperthermia on transport of low-density lipoprotein (LDL) through a stenosed arterial wall are analyzed comprehensively in the present work. The realistic and pertinent aspects of an arterial wall is represented by a multi-layer model, with a proper representation of the thickened intima region due to the atherosclerotic plaque formation. Effects of external and internal hyperthermia on LDL concentration levels are established along with the range of influence of these effects. Various modules of the current work are comprehensively compared with pertinent literature and are found to be in excellent agreement. The effects of external and internal hyperthermia as well as the load level and the axial location of the plaque formation on LDL transport and accumulation for a stenosed artery are established in this work.

With the rapid growth of data centers worldwide and the global shift towards energy sustainability, deploying new cooling technologies has an utmost importance. Conventional cooling systems such as chilled water system, usually have high capital costs and relatively low energy efficiency, leading to a high PUE and TCO values. Indirect evaporative cooling is a promising technology, which offers air cooling with high efficiency, hygiene air quality, and lower total cost. This paper details the design of a proof-of-concept data center with indirect evaporative cooling, which will be eventually deployed at megawatt-scale Baidu datacenters. BIN data analysis and CFD simulation are performed to optimize the physical design and operating conditions. CFD analysis of the data center room is established to optimize rack placement, air flow management, and cold aisle hot aisle configuration. A comprehensive TCO analysis is established, which shows a total savings of 9% using IDEC technology compared to chilled water system for cooling. In addition, TCO analysis indicates small to negligible effect of air supply temperature. Hence, air supply to the cold aisle is set to 27 °C to improve cooling performance. Finally, ROI sensitivity analysis is performed to measure the sensitivity of ROI on power usage effectiveness of the IDEC unit.

There is increasing evidence that renal impairment modifies nonrenal drug clearance through drug metabolizing cytochrome P450 (CYP) enzymes. In this study, the direct inhibitory effect of serum from chronic renal failure (CRF) patients receiving dialysis was evaluated in CYP3A4 (testosterone) and CYP2B6 (bupropion) metabolism assays. Human liver microsomes were incubated with ultrafiltered serum collected pre- and post-hemodialysis from ten CRF patients. Additionally, several uremic toxins were evaluated in the CYP3A4 assay. In only three patients was there a significant decrease or increase in testosterone or bupropion metabolism post-dialysis. Urea, mannitol, guanidine, homocysteine, uridine and creatinine had no effect on CYP3A4 metabolism. CMPF, hippuric acid and p-cresol had IC50 values that fell within CRF patient plasma concentrations. The IC50 values for indoxyl sulfate and indole-3-acetic acid were greater than CRF plasma concentrations. The lack of a consistent effect on CYP3A4 or CYP2B6 metabolism by uremic serum may be due in part to the frequency of hemodialysis in these patients which reduced the accumulation of uremic toxins. CMPF, hippuric acid and p-cresol have the ability to inhibit CYP3A4 metabolism at clinical concentrations which may correspond to reports of changes in hepatic metabolism in some CRF patients.

2538 Background: There is a high unmet medical need for effective treatments for patients with recurrent, metastatic, or persistent cervical cancer. Most patients are young and survival rates are poor. ORR for second line therapies is between 4 and 14% for chemotherapy and recently approved immunotherapy. Adoptive cell transfer using tumor infiltrating lymphocytes (TIL) have demonstrated durable responses in some patients with recurrent cervical cancer thus offering the potential for long-term disease control. Methods: Study C-145-04 is an ongoing, open-label, multicenter Phase 2 clinical trial evaluating the safety and efficacy of LN-145 TIL therapy in patients with advanced cervical cancer who have undergone at least one prior line of chemotherapy. Prior checkpoint inhibitor therapy is an exclusion criterion. The primary endpoint is ORR per RECIST 1.1; secondary endpoints include duration of response (DOR), disease control rate (DCR), and LN-145 safety. Tumors surgically harvested at local institutions are shipped to central GMP facilities for TIL generation in a 22-day manufacturing process. Final LN-145 TIL product is cryopreserved and shipped to sites. Patients receive one week of preconditioning lymphodepletion (cyclophosphamide, fludarabine), a single LN-145 infusion, followed by up to 6 doses of IL-2 (600,000 IU/kg). Results: As of 4 Feb 2019, 27 efficacy-c patients have received Gen 2 of LN-145, with a mean age of 47 years and 2.6 mean prior lines of therapy. Preliminary efficacy results: ORR was 44% (1 CR, 9 PR, 2 uPR), DCR was 89% at 3.5-month median study follow-up with 11/12 patients maintaining their response. Improved responses were observed in 4 patients with longer follow-up. Mean TIL cells infused was 28x109. Median IL-2 doses administered was 6.0. The adverse event profile was generally consistent with the underlying advanced disease and the profile of the lymphodepletion and IL-2 regimens. Conclusions: LN-145 results in 44% ORR in previously treated cervical cancer patients with acceptable safety and efficacy profile. LN-145 offers patients a viable therapeutic option warranting further investigation. Clinical trial information: NCT03108495.

Female age influence on male reproductive performance and also female fitness traits. The male recorded such as courtship activities, mating latency, copulation duration, and female fitness traits, such as, fecundity, has been used from isofemale lines of Drosophila melanogaster. The female age including, 2-3 days aged old (young aged), 17-18 days aged old (middle aged) and 31-32 days aged old (old aged) mated with virgin male (5-6 days aged old) that all of the flies were exposure of middle temperature (22°c). The results of SPSS software analyzed that in almost all of experimental young aged old had significantly level than middle and old age. Courtship activities elements, in young age female were greater than middle and old age, when mated with young age male. This variability suggests that parental age effected on the male reproductive performance and female fitness traits. Therefore, the female age specific possibility occurrence on reproductive access as follows by: young aged> middle aged>old aged.

The female age including, 2-3 days aged old (young aged), 17-18 days aged old (middle aged) and 31-32 days aged (old aged) mated with virgin male (5-6 days aged old). The results by SPSS software analayzed that in almost all of experimental young aged old had significantly level than middle and old age. Fecundity elements, in young age was greater than middle and old age, also the female ovariole elements when mated with male young age was more than middle and old age. This variability suggests that female age effected on the male reproductive performance and female fitness traits. Therefore, the female age specific possibility occurrence on reproductive access as follows by: young > middle >old aged.

A comprehensive analysis and optimization of a three-dimensional integrated circuit (3D IC) structure and its thermophysical attributes are presented in this work. The thermophysical and geometrical attributes studied in this paper include the die, device layer, heat sink, and heat spreader, which are critical structures within a 3D IC. The effect of the power density of the device layer which is the source of heat generation within the chip as well as the through silicon vias (TSV) and microbumps is also considered in our investigation. The thermophysical and geometrical parameters that have a significant impact on the thermal signature of the 3D IC as well as those that have an insignificant impact were established. The comprehensive analysis of different geometrical and thermophysical attributes can guide the design and optimization of a 3D IC structure and decrease the cost.

There is limited information concerning the brain's oxygen supply and demand in patients with sickle cell disease.We measured near-infrared spectroscopy of brain oxygenation in 27 patients with sickle cell disease regardless of vaso-occlusive crisis, 14 normal healthy controls, and five anaemic patients without sickle cell disease. We also measured pre- and post-transfusion cerebral oximetry in 14 additional sickle cell disease patients who were on transfusion programmes.The mean cerebral oxygen saturation in the combined steady-state and vaso-occlusive crisis population was found to be significantly lower than that in the controls and in anaemic patients without sickle cell disease (47.7% vs. 61.3%, 59.8%, P0.0001). Cerebral oxygen saturation failed to correlate with the haemoglobin concentration (r = 0.51, P0.5). However, cerebral oxygen saturation increased from 40.4% to 49.6% (P = 0.01) and correlated significantly with the haemoglobin level (r = 0.553, P = 0.003) in 14 subjects studied before and after transfusions. In seven subjects who received simple transfusions, cerebral oxygen saturation correlated strongly and positively with the haemoglobin level (r = 0.811, P = 0.001) and with percent normal haemoglobin (r = 0.786, P = 0.002), and negatively with abnormal sickle haemoglobin (r = -0.775, P = 0.003). None of these correlations was found to be statistically significant in the seven subjects given exchange transfusions. Cerebral oxygen saturation measured in the sickle cell disease subjects after transfusions was still significantly lower than in the anaemic subjects without sickle cell disease and in the normal controls (49.6% vs. 59.8% and 61.3%, P = 0.001).We found that patients with sickle cell disease have subnormal values of cerebral oxygen saturation. Red cell transfusions significantly increased the brain oxygenation in these patients. Cerebral oximetry may be a useful, noninvasive method for assessing the effect of circulating normal red cells in sickle cell patients after transfusions.

Hydroxyurea (HU), a chemotherapeutic agent, used increasingly in the treatment of sickle cell disease (SCD) stimulates the release of a tumor necrosis factor (TNF-alpha) from human macrophages in vitro and the concentration of TNF-alpha is greater than normal in subjects affected by SCD. It is widely accepted that HU may inhibit vaso-occlusive crisis (VOC) by stimulating the production of fetal hemoglobin (HbF) and nitric oxide (NO) in SCD; however, the beneficial effects of HU in vivo may be counteracted by the release of TNF-alpha and, in turn, the expression of a vascular cell adhesion molecule (VCAM-1) on leukocytes. Previous studies have shown that the severity of SCD increases with the leukocyte count. Therefore, we examined the relationship between plasma levels of TNF-alpha and HbF in SCD patients during steady-state (StSt) conditions (in the absence of VOC) and during VOC conditions after the acute administration of HU. Venous blood was collected in SCD patients over 6 h after administering a single dose of HU. Plasma TNF-alpha was found to be greater in SCD subjects than in reported normal adult controls (p0.05). TNF-alpha in the StSt group was not significantly different than in the VOC group; however, the plasma TNF-alpha tended to greater in the VOC group (p0.1). An increase in the HbF concentration after acute administration of HU (p0.01) was not associated with a significant change in plasma TNF-alpha (p0.1). Contrary to the results of in vitro studies, HU did not increase the plasma concentration of TNF-alpha. These findings suggest that a HU-induced increase in TNF-alpha does not contribute to VOC and sickle cell patients can be counseled that the HU-induced increase in TNF-alpha does not counteract the beneficial effects of HU in SCD.

Purpose The purpose of the study was to evaluate safety and determine the maximum tolerated dose (MTD) of MEDI-575, a fully human monoclonal antibody that selectively binds to platelet-derived growth factor receptor-α (PDGFRα), in patients with advanced solid tumors. Methods This phase I multicenter, open-label, single-arm study enrolled adults in a 3 + 3 dose escalation design to receive MEDI-575 (3, 6, 9, 12, or 15 mg/kg) once weekly (QW) until toxicity or disease progression occurred. One 0.5-mg/kg dose was given before the first dose in the 3-mg/kg cohort to determine pharmacokinetics (PK) and pharmacodynamics under unsaturated conditions. After completion of dose escalation in the QW cohorts, patients were enrolled in two additional cohorts and received MEDI-575 25 or 35 mg/kg every 3 weeks (Q3W). Secondary measures included assessments of PK, immunogenicity, and antitumor activity. Results A total of 35 patients received MEDI-575 QW (n = 23) or Q3W (n = 12). Most treatment-related adverse events were grade 1 or 2 in severity across all dose levels, with fatigue (n = 12) and nausea (n = 8) being reported most frequently. With no reports of dose-limiting toxicities (DLTs), the MTD was not reached. MEDI-575 exhibited a nonlinear PK profile and increased plasma platelet-derived growth factor-AA levels in a dose-dependent manner with limited immunogenicity. Stable disease was reported as the best tumor response in 9 of 29 evaluable patients; however, no objective responses were reported. Conclusion Administration of MEDI-575 QW or Q3W resulted in a favorable safety profile, including a lack of DLTs, but without evidence of antitumor activity in patients with refractory solid tumors.

The mechanism of action of hydroxyurea (HU) in decreasing the frequency of pain crisis in sickle cell disease (SCD) has not been fully elucidated. In vitro and in vivo studies suggest that nitric oxide (NO), a potent vasodilator, may partly be responsible for the beneficial effect of HU. This study was designed to determine the effect of oral administration of HU on plasma levels of NO metabolites (NO(x) ) in sickle cell patients (SCP). The results indicate that during steady-state plasma levels of NO(x) were significantly higher in HU-treated patients compared to non HU-treated patients or normal controls (p

TPS3096Background: The success of immune checkpoint inhibitors in various cancers has triggered the identification of novel approaches to modulate the immune system. CD73, an ectonucleotidase, cata...

TPS3095Background: Toll-like receptors (TLRs) are proteins in the innate immune system that, once bound to their ligands, directly activate innate immune cells, including myeloid dendritic cells, p...

Recent studies suggest that nitric oxide (NO) may partly be responsible for the beneficial effect of hydroxyurea (HU) in sickle cell disease (SCD) patients. NO stimulates cyclic guanosine monophosphate (cGMP) production, which mediates vasodilatation. We investigated the association between NO, cGMP and fetal haemoglobin (HbF) levels after HU administration. Our data showed that chronic HU significantly increased NO, cGMP, and HbF levels in SCD. Recently it was shown that HbF production was stimulated by cGMP-dependent protein kinase. Our results suggest that NO stimulates cGMP production, which then activates a protein kinase and increases the production of HbF.

Interactions between Bacillus anthracis (B. anthracis) and host cells are of particular interest given the implications of anthrax as a biological weapon. Inhaled B. anthracis endospores encounter alveolar macrophages as the first line of defense in the innate immune response. Yet, the consequences of this interaction remain unclear. We have demonstrated that B. anthracis uses arginase, inherent in the endospores, to reduce the ability of macrophages to produce nitric oxide ((•)NO) from inducible nitric oxide synthase (NOS2) by competing for L-arginine, producing L-ornithine at the expense of (•)NO. In the current study, we used genetically engineered B. anthracis endospores to evaluate the contribution of germination and the lethal toxin (LT) in mediating signaling pathways responsible for the induction of NOS2 and ornithine decarboxylase (ODC), which is the rate-limiting enzyme in the conversion of L-ornithine into polyamines. We found that induction of NOS2 and ODC expression in macrophages exposed to B. anthracis occurs through the activation of p38 and ERK1/2 MAP kinases, respectively. Optimal induction of NOS2 was observed following exposure to germination-competent endospores, whereas ODC induction occurred irrespective of the endospores' germination capabilities and was more prominent in macrophages exposed to endospores lacking LT. Our findings suggest that activation of kinase signaling cascades that determine macrophage defense responses against B. anthracis infection occurs through distinct mechanisms.

Vascular occlusive diseases affect brain blood flow, brain metabolism and are associated with arterial ischemic stroke. This study was designed to measure the brain blood flow velocity, brain oxygenation, hemoglobin concentrations, hematocrit, and cell free hemoglobin at pre- and post-exchange red cell transfusion in an 18 year old male patient with sickle cell disease and moyamoya syndrome (MMS). Exchange transfusion increased cerebral oxygen saturation 12%, total hemoglobin concentration 2%, hemoglobin AA 80%, and reduced sickle (SS) hemoglobin 12%, arterializations 33%, and cell free hemoglobin 33%. Brain blood flow velocity values were unaffected by transfusion. These observations suggest that exchange transfusion increases the hemoglobin carrying capacity and reduces sickle hemoglobin and shunting of blood, which may improve the peripheral and cerebral oxygenation. Transfusion did not affect the brain blood flow in this patient. Therefore the risk of transient ischemic attack and arterial ischemic stroke from mms still exist.

TPS3088 Background: Programmed cell death-1 (PD-1) is an inhibitory regulator or checkpoint of T-cell activation. Recent data from several large phase I studies have shown that targeting the PD-1/p...

TPS3087 Background: The PD-1/PD-L1 pathway plays a key role in controlling T-cell activation and may be utilized by tumor cells to evade antitumor responses. Anti-PD-1 and anti-PD-L1 antibodies hav...

In patients with sickle cell disease (SCD), cerebral oxygen saturation (rSO(2)) has been reported to be below normal and to increase after red blood cell transfusion.This study was designed to determine the effects of long-term and short-term hydroxyurea (HU) treatment on cerebral oxygenation in patients with SCD.This open-label pilot study was conducted at the Department of Anesthesiology and the Center for Sickle Cell Disease, College of Medicine, Howard University, Washington, DC. Adult African American outpatients with SCD and hemoglobin (Hb) genotype HbSS (homozygous sickle Hb) who were receiving long-term (6 months) HU treatment (15-30 mg/kg . d PO) or who had never received this treatment (control group) were enrolled. Patients in the treated and control groups were matched for age, sex, race, and Hb genotype. Cerebral oximetry (near-infrared spectroscopy) was performed to determine rSO(2) index. In a separate analysis to determine the effects of short-term HU treatment on cerebral oxygenation, hospitalized patients with SCD and vaso-occlusive crisis (VOC)receiving long-term therapy with HU were enrolled. We performed cerebral and pulse (fingernail) oximetry to determine rSO (2)index and arterial oxygen saturation (SpO(2)) after the administration of a single oral dose of HU (500-mg tablet) alone and again after dosing concomitantly with inhaled oxygen.The study enrolled 11 patients in the HU group (6 women, 5 men; mean [SD] age, 37 [8] years) and 20 controls (8 women, 12 men; mean [SD] age, 35 [6] years). Mean (SD) rSO(2) index was significantly increased (but still low) in patients receiving long-term HU treatment compared with controls (46.1% [6.6%] vs 41.2% [7.6%]; P0.025). Hb concentration (9.6 [1.4] g/dL vs 8.5 [1.2] g/dL; P0.027), hematocrit (28% [3%] vs 24% [4%]; P0.028), and mean corpuscular volume (102% [7%] vs 89% [8%]; P0.027) also were significantly higher in the HU group compared with controls. In 8 patients with SCD and VOC (6 men, 2 women; mean [SD] age, 28 [5] years), single-dose HU, either alone or in combination with inhaled oxygen, did not significantly affect cerebral oxygenation, and SpO(2) failed to correlate with rSO(2) index in these patients.The results of this open-label pilot study in patients with SCD suggest that the low cerebral oxygenation in these patients is significantly improved but not normalized with long-term HU treatment. A single dose of HU, either alone or in combination with inhaled oxygen, did not appear to influence cerebral oxygenation in patients with VOC.

These studies were designed as two experiments. Experiment 1 was performed to validate the hypothesis that oxygen saturation of the venous blood may be a marker for vaso-occlusive crisis (VOC) in sickle cell patients undergoing hydroxyurea (HU) treatments. Experiment 2 was performed to test the hypothesis that an acute increase in the blood nitric oxide (NO) concentration by administering HU modulates the perception of pain in sickle cell subjects in VOC. The percent saturations of oxyhemoglobin (%OPRE2/PREHb), reduced hemoblogin (%RHb), carboxy-hemoglobin (%COHb), met-hemoglobin (%MHb), fetal hemoglobin (HbF), and nitric oxide metabolites were measured in venous blood samples collected from sickle cell disease (SCD) who were on and off HU and OPRE2/PREat steady state and during VOC. The results showed the ratio of %OPRE2/PREHb/RHb in VOC+HU was significantly higher than patients in the steady state who were on and off of HU (p0.05). The %COHb was higher in all SCD groups, %COHb values were significantly different in SCD at steady state who were on HU. HU and OPRE2/PREtreatment did not play important role on venous blood %OPRE2/PREHb and pain scores in SCD during VOC. A single oral dose of HU was associated with a significant increase in the venous concentration of nitric oxide metabolites (NOx), p0.05. These findings suggest that the ratio %OPRE2/PREHb/RHb in venous blood and pain scores differentiate HU-untreated and HU-treated at steady state subjects from HU-treated subjects in VOC; however, the acute increase in venous NOx produced by administering HU to HU-treated subjects in VOC does not explain this difference.

Recent studies suggest that nitric oxide (NO) may partly be responsible for the beneficial effect of hydroxyurea (HU) in sickle cell disease (SCD) patients. NO stimulates cyclic guanosine monophosphate (cGMP) production, which mediates vasodilatation. We investigated the association between NO, cGMP and fetal haemoglobin (HbF) levels after HU administration. Our data showed that chronic HU significantly increased NO, cGMP, and HbF levels in SCD. Recently it was shown that HbF production was stimulated by cGMP-dependent protein kinase. Our results suggest that NO stimulates cGMP production, which then activates a protein kinase and increases the production of HbF.

3012 Background: MEDI3617 (M) is an investigational monoclonal antibody that inhibits angiogenesis by preventing the interaction of angiopoietin-2 (Ang2) ligands with the Tie2 receptor. Methods: Th...

The mechanism of action of hydroxyurea (HU) in decreasing the frequency of pain crisis in sickle cell disease (SCD) has not been fully elucidated. In vitro and in vivo studies suggest that nitric oxide (NO), a potent vasodilator, may partly be responsible for the beneficial effect of HU. This study was designed to determine the effect of oral administration of HU on plasma levels of NO metabolites (NO(x) ) in sickle cell patients (SCP). The results indicate that during steady-state plasma levels of NO(x) were significantly higher in HU-treated patients compared to non HU-treated patients or normal controls (p.05). In five inpatients in mild pain plasma levels of NO(x) increased significantly after 2 h of HU administration (p.05); however, in three inpatients in persistent pain with significantly lower baseline NO(x) there was a minimal NO(x) response to HU at 2 h (p.01). These observations indicate that HU administration is associated with the production of NO in some SCP, but that further study of the pharmacodynamics of this effect is necessary.

e18543 Background: Mesothelioma is a rare malignancy primarily attributed to asbestos exposure and has limited life expectancy. This study examined patient characteristics, and treatment patterns in commercially insured mesothelioma patients in real world setting. Methods: Patients aged ≥18 yrs with mesothelioma diagnosis were identified using ICD-9 and CPT codes/clinical algorithm between 7/1/2006 and 4/30/2011 from HealthCore Integrated Research Database. Patients with other cancers and without continuous insurance enrollment for ≤ 6-mos before or 12-mos post-diagnosis were excluded. Treatments were grouped into surgery, systemic chemotherapy (Ctx), and best supportive care (BSC) and were assessed descriptively. Results: A total of 184 patients were identified (mean age 64 yrs with 41% patients between 45 and 64 yrs old and 48% ≥65 yrs old, 64% males) with diagnosis of mesothelioma. 61% patients had pleural, 25% had peritoneal and 14% had pericardial disease. Only 10% patients received surgery, 27% received Ctx, while 50% received BSC (including radiation). Among patients who received 1st line Ctx (mean duration 90 days), almost half of patients received pemetrexed based regimen {pemetrexed + cisplatin (18%) or carboplatin (10%) and pemetrexed alone (20%)} followed by carboplatin monotherapy (42%). Among 41% who received 2nd line Ctx regimens (mean duration 105 days), most commonly used (65%) were pemetrexed based regimens {pemetrexed alone (35%) and pemetrexed + cisplatin (15%) or carboplatin (15%)} followed by single agent gemcitabine (15%). No patients received vinorelbine (0%). In patients who received Ctx, 35% died before receiving 2nd line Ctx and an additional 15% of patients died before receiving 3rd line Ctx. Only 16% received 3rdline Ctx. Around 41% patients had at least one emergency room visit and 80% had at least one hospitalization. Conclusions: Mesothelioma is predominant in males and older adults in commercially insured patients in the United States. Systemic chemotherapy remains the main treatment strategy with pemetrexed based therapy being most commonly used regimen. This study highlights limited treatment options and high unmet need in this population.

e13075 Background: 153Sm and ZA are often employed to manage complications related to BM. Although 153Sm and ZA bind to overlapping sites within the bone, there is a paucity of information on potential pharmacokinetic (PK) and pharmacodynamic interactions between the two. In this study we report the pharmacokinetics of 153Sm when administered in combination with ZA. Methods: The study was carried out as two independent phase I trials assessing the effects of escalating dose 153Sm (0.5, 0.75, 1.0, and 1.25 µCi/kg) administered prior to or immediately after intravenous ZA 4 mg. Study 1 recruited 12 ZA-naive PC pts, while Study 2 enrolled 10 PC pts who had received at least 3 months of ZA prior to trial entry. On study, pts received 2 doses of 153Sm given 3 months apart and 6 doses of ZA given monthly. In Cycle 1 of both trials, 153Sm was administered on day1 and ZA on day 2. In Cycle 2, ZA was administered 1 hour prior to 153Sm on the same day. Blood samples for radioactive analysis were obtained at 5 min, and 0.5, 1, 2, 4, and 24 h post 153Sm dosing. PK analysis was performed by both model independent and PK modeling methods using WinNonlin. Results: The PK parameters of 153 Sm were highly variable between patients. The coefficient of variation of the Cmax/Dose was greater than 50% in all subgroups. A dose related increase in exposure to 153Sm was observed with high variability as well. The estimated terminal half-life (t1/2) was similar across all dose levels within each study. The t1/2 for Cycle 1 was 4.73 ± 1.27 for Study 1 and 6.73 ± 3.92 h for Study 2. The dose normalized Cmax (Cmax/Dose) was 0.068 ± 0.036 and 0.061 ± 0.043 µCi/mL for Study 1 and Study 2, respectively. Pharmacokinetic parameters for Cycle 2 were similar to those calculated for Cycle 1 within each study. Conclusions: The Cmax and t1/2 reported here are similar to previous studies of 153Sm with doses ranging from 1.0 to 3.0 µCi/kg in ZA-untreated pts. Although not statistically significant, the subtle differences in the pharmacokinetics of 153Sm merit further investigation.

Recently we reported the use of a non-invasive near-infrared optical spectroscopy technique to measure cerebral oxygenation (cerebral oximetry, rSO2%) in agroup of 27 adult patients with sickle cell disease (Eur J Clin Invest, 34:143,2004). The sickle cell patients’ rSO2 values were significantly lower (mean 47.7%) than those in normal subjects (mean 61.3%) even though none of the patients had clinical evidence of stroke or cerebral ischemia. We included patients with all Hb phenotypes, and regardless of hydroxyurea (HU) treatment. Transfusions improved cerebral oxygen saturation but the post-transfusion values still did not reach normal levels. Our findings were corroborated independently by Raj et al. who studied 25 children with sickle cell disease (J Pediat Hematol Oncol 26:279,2004). In order to determine if long-term HU treatment affects rSO2, we analyzed cerebral oximetry results in a subset of 31 patients with sickle cell anemia (Hb SS). Eleven of them were on long-term (more than 6 months) HU treatment at stable doses (1000–1500 mg/day). The table shows that the mean rSO2, Hb, Hct, and MCV in HU-treated patients were significantly higher than those in sickle cell anemia (SCA) patients not on HU. The rSO2 in HU-treated patients was 12.5% higher than in SS patients not on this drug. By comparison, we previously reported a 24% increment in rSO2 following transfusions. A group of 8 patients who were on long-term HU treatment were given also single 1000 mg oral doses of HU and their rSO2 was measured for 12 hours without noticeable change in cerebral oxygenation. Nor did rSO2 change after oxygen inhalation (3L/min). The cause of the low rSO2 in sickle cell patients is unknown and still under investigation. It is probably not related exclusively to the anemia, since, as previously reported, anemic subjects without sickle cell disease appear to have normal rSO2. These preliminary results indicate that chronic HU treatment is associated with higher rSO2 values in SCA. If validated in a larger number of patients, our findings suggest that cerebral oximetry could be a useful, non-invasive method for assessing a new in vivo effect of HU and red cell transfusion in sickle cell disease: increased blood oxygen saturation in the cerebral vasculature. HYDROXYUREA AND CEREBRAL OXYGEN SATURATION IN PATIENTS WITH SICKLE CELL DISEASE | | NO HYDROXYUREA (N=20) | HYDROXYUREA (N=11) | P value* | |:-----------------------------------------------------------------------------:| --------------------- | ------------------ | -------- | | rSO2 = cerebral oxygen saturation. *= t-test. Plus/minus figures represent SD | | Mean rSO2 (%) | 41 ± 6.6 | 46 ± 7.6 | 0.025 | | Mean Hb (g/dl) | 8.4 ± 1.4 | 9.68 ± 1.2 | 0.029 | | Mean Hct (%) | 24± 3.4 | 28± 4.4 | 0.027 | | Mean MCV (fl) | 89± 8 | 102± 7 | 0.028

Meeting abstracts Despite improvements in diagnosis, surgical techniques, and multidisciplinary approaches to patient care, the median survival of patients with metastatic gastroesophageal adenocarcinoma is less than one year. Chemotherapy is the mainstay of treatment in patients with metastatic