Time Point Of Administration Research Articles

The earlier, the better: the effects of different administration timepoints of sorafenib in suppressing the carcinogenesis of VEGF in rats

PurposeTo investigate the optimal starting time point of sorafenib therapy in suppressing the tumor-promoting effects of VEGF up-regulation, which is frequently found after local therapy in clinical practice.MethodsVEGF was intravenously injected to imitate the evaluated expression after local tumor therapy, such as TACE. A total of 40 SD rats bearing hepatic tumors were randomly divided into four groups and sorafenib was administered at different timepoints: (A) control group: VEGF injection only; (B) initiating sorafenib 72 h prior to VEGF injection; (C) initiating sorafenib simultaneously with VEGF injection; (D) initiating sorafenib 72 h post-VEGF injection. The rate of tumor growth, median survival time, expression of VEGF, and microvessel density (MVD), as determined by immunohistochemical (IHC) examination, were compared.ResultsThe results revealed that the tumor size and median survival time were significantly different between the three sorafenib groups compared to the control group (p < 0.05). Median survival times were 19.6 ± 1.78, 31.2 ± 6.99, 27.4 ± 4.9, and 26.5 ± 4.6 days in group A, B, C, and D, respectively. Furthermore, there was a difference in statistical significance between the two sorafenib groups B and D (p = 0.04). Tumors were collected for HE staining and IHC examination. The expression levels of VEGF in B, C, and D were 42.8 ± 7.96, 71.9 ± 15.73, and 73.6 ± 13.73, and all of them were significantly lower than that in the control group (88.3 ± 13.61). Furthermore, the level of MVD was 109.2 ± 8.98 in the control group, which was significantly higher than in the three sorafenib groups (45.7 ± 16.92, 77.1 ± 16.29, and 93.6 ± 12.87, all p < 0.05).ConclusionsAccording to our results, the most suitable regimen for the administration of sorafenib is before the increased expression of VEGF, which showed a potential advantage for controlling the tumor growth and prolonging the survival time of test animal via inhibiting VEGF-receptor expression through the bifunction of VEGF, and the reduction of tumor angiogenesis.

Sequential Delivery of Ionizing Radiation Mitigators Based on Plasma, Intestine, and Bone Marrow Protein Signatures

Total body irradiation (TBI) triggers acute injury of sensitive cell populations in the bone marrow and the intestine thus triggering a cascade of delayed damage responses causing massive cell death through diversified programmed mechanisms and penultimately leading to multiple organs failure and lethality. Based on this principle, the radiomitigative strategies should include precisely timed treatments with specific regulators of individual pathways dominating the pathogenic process at this particular stage. Both JP4-039, an anti-apoptotic GS-nitroxide (Rwigema, J.C., et al., Int. J Radiat Oncol Biol Phys, 80(3):860-868, 2011), and Necrostatin-1, an inhibitor of necroptosis (Huang Z., et al., Biochemica at Biophysica Acta, 1862:850-856, 2016), have been shown to be effective irradiation mitigators. We determined whether proteins detectably altered in the plasma, bone marrow, and intestine of total body irradiated mice could be used as signatures to direct sequenced administration of two radiation mitigators with distinct mechanisms of action. C57BL/6NTac adult female mice were irradiated to the LD50/30 dose of 9.25 Gy TBI, and were evaluated daily over 7 days in 4 groups: 1) Radiation Only; 2) 9.25 Gy + JP4-039 at 24 hrs post-irradiation; 3) 9.25 Gy + Necrostatin-1 at 48 hrs post-irradiation; and 4) 9.25 Gy + both JP4-039 at 24 hrs post-irradiation + Necrostatin-1 at a second delayed time point post-irradiation. Luminex immunoassays were run on plasma, intestine, and bone marrow for each group (n=4-5 each data point) at serial time points at day 0 (prior to irradiation), 1, 2, 3, 4, 5, 6, or 7 after TBI. We measured 33 proteins known to be associated with response of tissues to ionizing irradiation. Data was used to generate “heat maps” of all 33 proteins to compare each experimental group with radiation only control. Survival studies were carried out for each radiation mitigator combination to determine the best time point for sequential administration of the second radiation mitigator (n=15 for each group). Administration of JP4-039 at 24 hrs post-irradiation (I.V. 20 mg/kg in 100 μl F14 emulsion) delayed the elevation of plasma IL-6 to Day 3 which correlated with the time of increased bone marrow IL-1α signature for the onset of necroptosis. Increased survival was observed after JP4-039 at 24 hrs and when Necrostatin-1 was delayed until 72 hrs post-irradiation. This increased survival correlated with an increase in levels of plasma proteins including G-CSF, IL-6, KC, and eotaxin, which are indicators of stimulated cell proliferation and chemotaxis. Plasma levels of proteins did not always match changes in the bone marrow or intestine. Sequential administration of JP4-039 at 24 hrs and Necrostatin-1 at 72 hrs post-irradiation increased survival, and correlated with increased levels of plasma inflammatory proteins. Elevations in levels of some plasma proteins may reflect irradiation effects on organs other than bone marrow or intestine.

Efficacy and safety of sugammadex versus neostigmine in reversing neuromuscular blockade in adults.

Efficacy and safety of sugammadex versus neostigmine in reversing neuromuscular blockade in adults.

Anti-vascular endothelial growth factor treatment decreases bladder pain in cyclophosphamide cystitis: a Multidisciplinary Approach to the Study of Chronic Pelvic Pain (MAPP) Research Network animal model study.

To investigate whether treatment with anti-vascular endothelial growth factor (VEGF)-neutralizing antibodies can reduce pain and voiding dysfunction in the cyclophosphamide (CYP) cystitis model of bladder pain in mice. Adult female mice received anti-VEGF-neutralizing antibodies (10mg/kg i.p. B20-4.1.1 VEGF mAb) or saline (control) pre-treatment, followed by CYP (150mg/kg i.p.) to induce acute cystitis. Pelvic nociceptive responses were assessed by applying von Frey filaments to the pelvic area. Spontaneous micturition was assessed using the void spot assay. Systemic anti-VEGF-neutralizing antibody treatment significantly reduced the pelvic nociceptive response to CYP cystitis compared with control (saline). In the anti-VEGF pre-treatment group, there was a significant increase in pelvic hypersensitivity, measured by the area under the curve (AUC) using von Frey filaments at 5h post-CYP administration (P = 0.004); however, by 48h and 96h post-CYP administration, pelvic hypersensitivity had reduced by 54% and 47%, respectively, compared with the 5h post-CYP administration time point, and were no longer significantly different from baseline (P = 0.22 and 0.17, respectively). There was no difference in urinary frequency and mean voided volume between the two pre-treatment groups. Systemic blockade of VEGF signalling with anti-VEGF-neutralizing antibodies was effective in reducing pelvic/bladder pain in the CYP cystitis model of bladder pain. Our data support the further investigation of the use of anti-VEGF antibodies to manage bladder pain or visceral pain.

Identifying Students at Risk: An Examination of Computer-Adaptive Measures and Latent Class Growth Analysis

ABSTRACTMultitiered systems of support depend on screening technology to identify students at risk. The purpose of this study was to examine the use of a computer-adaptive test and latent class growth analysis (LCGA) to identify students at risk in reading with focus on the use of this methodology to characterize student performance in screening. Participants included 3,699 students in Grades 3–5. Three time points of administration (fall, winter, and spring) of the computer-adaptive reading measure were selected. LCGA results indicated 6–7 classes, depending on grade, informed by level and growth in student performance that significantly predicted failure on the statewide test administered at the end of the year. The lowest-performing classes had failure rates above 90% across all grades. The results indicate that identifying homogeneous groups of learners through LCGA may be valuable as an approach to determining students who need additional instruction. Practical implications and future directions are discussed.

Critical time point for apoptotic cell death in an experimental ischemia/reperfusion model and the effect of N-acetylcystein

Abstract Objective Kidney transplantation is an important treatment option in end stage renal failure. Tissue death may be an important problem when a kidney is removed from a cadaver and transported to a donor a long distance away. The purpose of this study is to determine the critical time point for apoptotic cell death in a renal ischemia/reperfusion model and determine the effects of N-acetylcystein on apoptosis induced by ischemia injury. Methods Apoptotic cell death after induced renal ischemia followed by reperfusion, was estimated in a group of Wistar albino rats by immunoflourescence and ELISA techniques. N-acetylcystein, an antioxidant agent, was given to the rats to study the effect on apoptosis. Tissues were examined immunohistochemically at 0, 1 h, 24 h, 5 days and 10 days for detection of apoptotic cells. Results Our results showed that an ischemia for 60 min followed by reperfusion for 60 min triggered apoptosis. Moreover, N-acetylcystein significantly diminished both the ischemia/reperfusion damage and apoptosis. Conclusion We anticipate our results would be important for kidney transplantation in estimating the critical time point for apoptosis and administration of N-acetylcystein prior to removal of the organ may be important in delaying the onset of apoptosis.

Abstract A28: Imaged-based computational predictions of imaging agent efficacy in pancreatic tumors expressing TLR2

Abstract The current therapies continuously fail to provide successful results for pancreatic adenocarcinoma, one of the most deadly cancers with only 6% overall 5-year survival rate. The improvement of techniques for early detection, predicting of treatment efficacy and monitoring of tumor response during and after therapy remain under focused research. By combining computational modeling with intravital fluorescence microscopy we developed a novel method to assess intratumoral and intracellular distribution of imaging agents targeted to pancreatic cancer cells. Our long-standing goal is to provide a tool for optimizing efficacy of targeted agents based on individual patient data. Our team reported previously that, in addition to the important role the toll-like receptor 2 (TLR2) plays in the immune system response, TLR2 is also a bona fide cell-surface marker for targeting pancreatic cancer cells. TLR2 recognizes a vast number of biomolecules, including lipopeptides, such as a novel TLR2 ligand designed in our laboratory (TLR2L). We used intravital fluorescence microscopy methods for the real time in vivo imaging of TLR2L conjugated to near-infrared fluorescent dye, Cyanine 5 (TLR2L-Cy5), and its distribution in the tissue of pancreatic adenocarcinoma tumor xenografts in mice with endogenous expression of TLR2. In order to quantify the space- and time-dependent dynamics of TLR2L we combined intravital dorsal window chamber experiments with computational modeling of TLR2L-Cy5 diffusion and internalization following its intravenous administration. Our computational model is calibrated to the tissue histology of a particular tumor and accounts for explicitly defined individual tumor cells and expression of cell membrane receptors, as well as the extracellular matrix interpenetrated by the interstitial fluid, and tumor vasculature. The fluorescent imaging agent is also modeled as individual discrete molecules that extravasate via influx from blood capillaries, move through the tumor and become internalized by the cells. Our microscopic level computational model allowed for quantitative assessment of targeted imaging agent extravasation, interstitial diffusion and intracellular accumulation on a cell-to-cell basis. When accounting for various values of diffusion and binding affinity, we identified a cross-dependence between these physical characteristics of targeted agents. Our computational results revealed that agents of distinct affinities can reach similar efficacies of binding and internalization. Moreover, we identified the optimized dose-response protocols appropriate for any targeted agent with a priori known physical and biochemical properties. Finally, when considering different extravasation schemes of the agent we also showed that the time of crossing the capillary wall by the ligand plays an important role in effective binding and saturation of cell receptors and intracellular distribution. In addition to improvement in drug delivery schemes, our model predicts which biochemical and physical properties of targeted agents could be tuned for the maximum effect in patient-specific extracellular matrix environments, tumor topologies and receptor expression levels. We present an interdisciplinary approach to quantify diffusion and cellular uptake of an imaging agent targeted to pancreatic cancer cell lines expressing the TLR2 receptor. This integrated method can be used for optimizing the administration schedules and time points for data collection from individual human tumor xenografts in order to personalize the treatment and improve its efficacy. In the future, the model can be extended other targeted imaging or therapeutic agents, to other solid tumors, and to an individual patient's tumor histology. Citation Format: Aleksandra Karolak, Veronica Estrella, Tingan Chen, Amanda Huynh, David Morse, Katarzyna Rejniak. Imaged-based computational predictions of imaging agent efficacy in pancreatic tumors expressing TLR2. [abstract]. In: Proceedings of the AACR Special Conference on Engineering and Physical Sciences in Oncology; 2016 Jun 25-28; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2017;77(2 Suppl):Abstract nr A28.

A comparison in therapeutic efficacy of several time points of intravenous StemBell administration in a rat model of acute myocardial infarction

BackgroundAdipose-derived stromal cells (ASCs) are a promising new therapeutic option for patients with acute myocardial infarction (AMI). Previously, we found that ASCs coupled to antibody-targeted microbubbles (StemBells [StBs]) improved cardiac function when administered intravenously 7 days post-AMI in rats. In this study, we compared the efficacy of intravenous StB administration at different administration time points following AMI in rats. MethodsAMI, followed by reperfusion, was induced in four groups of male Wistar rats, which subsequently received an intravenous 1 × 106 StB bolus 1 day post-AMI (StB1; n = 8), 7 days post-AMI (StB7; n = 9), at both time points (StB1+7; n = 7) or neither (Control; n = 7). The effect onrdiac function was determined using echocardiography prior to AMI, 7 days post-AMI and 42 days post-AMI. The effect on infarct size and macrophages in the infarct core were determined (immuno)histochemically 42 days post-AMI. ResultsAt 42 days post-AMI, all three StB groups had a significantly improved fractional shortening compared with the control group. Between the StB-treated groups, the effects did not differ significantly at 42 days post-AMI. At 7 days post-AMI, the StB1 group had a significantly improved fractional shortening compared with the control and StB7 groups. No significant changes in infarct size or macrophage numbers were found compared with the control group for any StB group. ConclusionsStB administration resulted in long-term improvement of cardiac function, independent of the time point of administration. When administered at 1 day post-AMI, this improvement was already evident at 7 days post-AMI.

The Impact of a Practice-Teaching Professional Development Model on Teachers’ Inquiry Instruction and Inquiry Efficacy Beliefs

This study examined changes in middle school teachers’ beliefs about inquiry, implementation of inquiry practices, and self-efficacy to teach science through inquiry after participating in a year-long professional development program. The professional development model design was based on Bandura’s (1986) social cognitive theory of learning and literature on effective professional development. Using the Electronic Quality of Inquiry Protocol (EQUIP) before and after the program (Marshall, Smart & Horton, 2010), teachers’ quality of inquiry instruction significantly increased across all 4 EQUIP factors. To assess teachers’ inquiry teaching efficacy, the Teaching Science as Inquiry (TSI; Dira-Smolleck, 2004) instrument was administered 3 times to the teacher participants. Changes between time points of TSI administration were examined for personal self-efficacy and outcome expectancy across 5 essential features of classroom inquiry. Findings for the personal self-efficacy scale showed statistically significant gains from pre-Institute to 1-year follow-up for all 5 inquiry features. Teachers’ outcome expectancy beliefs significantly increased across the program for 3 of the 5 essential features of inquiry. The results indicate the effectiveness of the professional development model at increasing the participants’ inquiry efficacy as well as the quality of their inquiry instruction. Our study provides evidence to support the need to include collaborative practice-teaching and reflection opportunities within professional development models that aim to increase teachers’ use of inquiry-based instructional strategies.

Timing of predisposing factors is important in necrotic enteritis models

ABSTRACTSince the ban of antimicrobial growth promotors, the importance of necrotic enteritis in broilers increases. Reliable and reproducible infection models are required for pathogenesis studies and product screening. Two major predisposing factors in necrotic enteritis models are fishmeal supplementation to feed and Eimeria infection. However, many unsolved issues regarding these predisposing factors still exist. Therefore, the influence of timepoint of fishmeal administration (onset on day 8 or day 18), timing of coccidiosis challenge (day 15 or day 19) and strain of coccidiosis challenge (field strain vs. commercial vaccine) on the induction of necrotic enteritis lesions was investigated. The birds were inoculated with Clostridium perfringens three times per day for four consecutive days (day 17 until day 20) and were scored for the presence of necrotic enteritis on days 22, 23, 24, 25 and 26. Supplementation of the diet with fishmeal from day 8 onwards increased the likelihood of necrotic enteritis compared to supplementation from day 18 onwards. Birds challenged on day 19 with coccidiosis were more likely to have necrotic enteritis on scoring days 23 and 24 compared to birds challenged on day 15. Differences on other scoring days were less pronounced. Finally, the strain of coccidiosis challenge had little influence on the induction of necrotic enteritis. Findings of this study can help researchers to set up successful necrotic enteritis infection models.

Abstract B21: Using computational modeling to quantify targeted agent binding and internalization in pancreatic cancers

Abstract The pancreatic adenocarcinoma is one of the most deadly cancers with only 6% overall 5-year survival rate. Since the current therapies fail to provide successful results, the improvement of techniques for early detection, predicting of treatment efficacy and monitoring of tumor spread during and after surgical procedures remain under focused research. The purpose of this study is to develop a novel method to assess behavior (diffusion and binding within the tissues) of imaging agents targeted to pancreatic cancer cells. Our long-term goal is to design a prospective tool for the prediction of targeted imaging agent peak concentration and cellular uptake based on individual patient data. The toll-like receptor 2 (TLR2) is known to play an important role in the immune system response. In addition, our team reported that TLR2 is a bona fide cell-surface marker for targeting pancreatic cancer. TLR2 recognizes a vast number of biomolecules, including lipoproteins, such as designed by our team, novel TLR2 ligand (TLR2L). Recent development of an intravital fluorescence microscopy method allowed for the real time in vivo imaging of the TLR2L conjugated to near-infrared fluorescent dye, Cyanine 5 (TLR2L-Cy5) and its penetration through the tissue of pancreatic adenocarcinoma tumor xenografts in mice with endogenous expression of TLR2. In order to quantify the space- and time-dependent dynamics of TLR2L we combined intravital dorsal window chamber experiments with computational modeling of TLR2L-Cy5 diffusion and internalization following intravenous administration. Our computational model accounts for an explicitly defined tissue morphology composed of individual tumor cells, extracellular matrix interpenetrated by the interstitial fluid, and tumor vasculature. We also model individual molecules of a fluorescent imaging agent that extravasate via transmural influx from blood capillaries, spread through the tumor and become internalized by the cells. The process of agonist binding to the receptor is modeled by trapping the particle by the near-by cell boundary receptor, which mimics the distance dependent electrostatic and hydrophobic interactions initiating the recognition and binding in vivo. Our studies led to quantification of the TLR2L-Cy5 intratumoral transport including agent extravasation, diffusion and intracellular accumulation in relation to tumor tissue structure and vascular architecture. The performed computational simulations allowed for detailed continuous in time assessment of the targeted imaging agent penetration on the microscopic (cell-to-tissue) level. The calibrated in silico model revealed the time-dependent dynamics of TLR2L-Cy5 agent binding, internalization and intracellular distribution. Our quantitative approach also showed a non-uniform spatial saturation of the TLR2L-Cy5 on the plasma membrane and inside the cell. Moreover, continuous simulations allowed for prediction of the maximum concentration peak in the extracellular matrix and for the uptake peak in individual cells. In addition to quantitative analysis, we provide novel (two- and three-dimensional) visualization tools for imaging agent dynamical penetration, membrane binding and receptor-mediated endocytosis process in pancreatic cells. In conclusion, we present an interdisciplinary approach to quantify diffusion and cellular uptake of an imaging agent targeted to pancreatic cancer cell lines expressing the TLR2 receptor. This integrated approach can be used in the future for the development of other targeted imaging and therapeutic agents, for other solid tumors, and for optimizing the administration schedules and time points for data collection from individual human tumor xenografts in order to improve treatment efficacy. Citation Format: Aleksandra Karolak, Veronica Estrella, Tingan Chen, Amanda Huynh, David Morse, Katarzyna Rejniak. Using computational modeling to quantify targeted agent binding and internalization in pancreatic cancers. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr B21.

Thyroid hormone modulates neuroglobin and cytoglobin in rat brain

Thyroid hormones (THs) are essential and crucial for brain development, playing a role in growth and differentiation. Two globins named neuroglobin (Ngb) and cytoglobin (Cygb) are located in the brain, and each one has different distribution and function: They seem to have similar action by providing O(2) for respiratory chain, and detoxification of reactive oxygen species (ROS) and nitric oxide (NO) protecting tissues against irreversible lesions. We aimed to investigate the influence of thyroid state in Ngb and Cygb metabolism in different brain regions and evaluate their responses in cerebellum, hippocampus and cerebral cortex (hereafter called as cortex) after supraphysiological doses at different time points of TH administration. Experiments were carried out in rats, divided in eight experimental groups Control (C), thyroidectomy (Tx), and thyroidectomy treated with jugular intravenous injection (i.v). T3 (100 μl/100 g) injection and sacrificed after 30, 60, 120 min and 6, 12 and 24 h. In cortex, we found increase in Ngb gene and protein expression in different time points compared to C group, however Cygb gene and protein expression were decreased. In hippocampus, Ngb and Cygb protein expression increased 24 h after i.v. T3 injection in comparison to Tx. In cerebellum, we found increased Ngb gene expression after 120 min, 6, 12 and 24 h after T3 administration compared to Tx, and in contrast, protein expression was found to be significantly increased only 12 and 24 h compared to Tx. Ngb and Cygb expression in brain is influenced by thyroid hormone state both by its lack or excess.

Quantitative Analysis of Small Molecular Weight Thiols and Disulfides in Blood from a Sickle Cell Disease Patient Infused with N-Acetyl-L-Cysteine

N-acetyl-L-cysteine (NAC) is an FDA approved drug used to treat acetaminophen overdose or as a mucolytic agent in respiratory disorders. The commonly accepted mechanism of action is that NAC undergoes deacetylation to cysteine, which is then used to synthesize glutathione (GSH), a major intracellular antioxidant. Like other thiol-containing compounds, NAC can also act as a reducing agent to break protein disulfide bonds or as a scavenger of reactive oxygen species. Due to its antioxidant properties, NAC has been proposed as a potential treatment for many diseases associated with oxidative stress, including sickle cell disease (SCD), neurological disorders, infectious diseases, and cancers. Though NAC has been widely studied, a full understanding of the mechanism by which NAC is effective in vivo has been limited by challenges in accurately quantifying NAC and its metabolites.As part of a clinical trial of NAC therapy in SCD, we have developed a liquid chromatography-mass spectrometry (LC-MS) based assay to quantify small molecule free thiols and disulfides using isotopically labeled internal standards. We applied this method to quantify small molecular thiols and disulfides in whole blood, red blood cells, and plasma from a SCD patient before (pre) and at 1, 8, 24 and 72 hr time points of intravenous administration of NAC at a dose of 300 mg/kg (a bolus infusion of 150 mg/kg for 1 hour followed by 150 mg/kg given over the next 7 hr). The cysteine concentration in whole blood increased to 286 μM at 1 hr from 97 μM at baseline, indicating that NAC is indeed rapidly metabolized (deacetylated) to cysteine. Interestingly, although cysteine concentration in RBCs increased over 4 fold at 1 hr and remained high compared to baseline, the highest concentration of total GSH in blood was observed at 24 hr (743 μM compared to 494 μM at baseline). Intracellular availability of cysteine is known as a rate-limiting step for GSH synthesis, and the delayed accumulation of GSH may suggest that NAC is involved in the extracellular deficit of reducing equivalents before it serves as a substrate in GSH synthesis.To explore this possibility, we quantitated NAC and its oxidation products, homo- and mixed disulfides. We found that total NAC concentration reached 1.58 mM in whole blood at 1 hr, but 44% of NAC was oxidized to N-acetyl-cystine (NAC-ss) or formed mixed disulfides with GSH (GS-ss-NAC) and Cys (Cys-ss-NAC), whereas the NAC used for infusion contained less than 0.5% in the oxidized form (NAC-ss). Concurrent with the formation of NAC disulfides, the levels of oxidized GSH (GSSG, GS-ss-Cys) and cysteine (cystine) were significantly decreased. These observations suggest that NAC administration of SCD not only increases GSH levels by raising the cysteine concentration, but also directly functions as an antioxidant to reduce oxidative stress. SCD patients are known to have low levels of GSH and frequently experience oxidative stress. NAC treatment is likely to address both issues. We plan to analyze the effects of NAC on blood small molecule thiol concentrations in several more SCD patients. DisclosuresNo relevant conflicts of interest to declare.

Commentary: Different immunological mechanisms govern protection from experimental stroke in young and older mice with recombinant TCR ligand therapy.

Translational research to discover therapeutic targets for stroke has not fared well in developing new treatments for patients. The three most pressing issues are the choice of therapeutic target, animal model, and timepoint of administration of treatment. This manuscript “Different immunological mechanisms govern protection from experimental stroke in young and older mice with recombinant TCR ligand therapy” provides a fresh approach toward developing stroke treatments (Offner et al., 2014). This study examines treating with recombinant TCR ligand (RTL) in both young and older mice 4 hours after MCAO. RTL consist of the specific domains of MHC II molecule and inhibits activation of T cells toward inflammation. The novelties of this study are several fold including: (1) inhibition of a specific immunoinflammatory pathway to avoid total immunosuppression, (2) use of elderly mice to mimic the population of human patients and (3) administration at a clinically relevant timepoint. For a number of years, researchers have reported the entry of immune cells into the area of the infarct, which leads to further neurodegeneration. However, targeting this immune response as a treatment for stroke remains elusive (Iadecola and Anrather, 2011). In the past several years, the spleen has been reported to be a focal point for the immune system to mount an inflammatory response that exacerbates stroke-induced neurodegeneration (Offner et al., 2006; Vendrame et al., 2006). In fact, ablation of the spleen reduces inflammation and neural cell death in the rodent brain after experimental stroke (Ajmo et al., 2008; Ostrowski et al., 2012; Jin et al., 2013). Blockade of interferon gamma signaling has been reported to be neuroprotective in experimental stroke (Liesz et al., 2009, 2011; Seifert et al., 2014) although other groups have reported alternative results in their model systems (Chu et al., 2000; Lambertsen et al., 2004). T cells are the main effector cell of this neurodegenerative response and responsible for the release of the proinflammatory interferon gamma, which is a potent activator of microglia becoming neurotoxic (Boehm et al., 1997; Mebius and Kraal, 2005). Moreover, addition of interferon gamma reverses the neuroprotection provided by splenectomy, demonstrating that this cytokine plays a major role in the spleen eliciting response resulting in further neural death after stroke (Seifert et al., 2012). As shown in this study, administration of RTL directly inhibits the T cell response to stroke, which concomitantly deactivates the splenic response as well in both young and old mice. This treatment reduced infarct volumes in both age groups by altering the immune response to this neurological insult. Most interestingly, the recombinant TCR ligand differentially affected components of the immune response in the elderly and young mice but still resulted in a blunted immune response reducing neurodegeneration. This study illuminated differences in the immune cell composition and inflammatory expression between these two age groups in response to stroke. This is one of the few studies that enlists elderly animals which begins to provide insight into the differences in the physiological responses to stroke between young and the aged animals. Such differences in young and elderly could be responsible for the failure to translate findings at the bench to those in the clinic since the vast majority of studies use greatly cheaper young ones. More studies using elderly rodents will further supply additional insight in translating stroke treatment from the preclinical research to the clinical setting. This study sets the basis for future ones to develop new therapeutic approaches for a treatment for stroke.

Optimizing Performance of Endogenous Neural Stem Cell Therapy for Ischemic Stroke: A Neuroinformatics and Neuroimaging Approach to Translational Medicine

Event Abstract Back to Event Optimizing Performance of Endogenous Neural Stem Cell Therapy for Ischemic Stroke: A Neuroinformatics and Neuroimaging Approach to Translational Medicine Suhela Kapoor1*, VPS Rallabandi1 and Prasun Roy1 1 National Brain Research Centre, India Aim of study: To design an in-silico model to develop a translational-medicine basis of endogenous neural stem-cell-based therapy for ischemic stroke using pharmacological growth factors and by utilizing neuroimaging tools with in-vivo verification to translate the results to patients. Most stroke patients incur residual disability despite treatment, thus needing newer approaches of neurorestoration/regenerative medicine. In our study, the phenomena of neurogenesis, synaptogenesis and progenitor cell migration from the sub-ventricular zone of the brain towards the penumbra surrounding the ischemic lesion, under influence of therapeutic growth factor introduced interventionally, were employed as model parameters. Material & Methods: A predictive mathematical model was designed to discretize the steps involved in neural stem cell proliferation, migration and differentiation leading to neurorestorative recovery. MATLAB algorithms were run to compute the optimal dosage and time-point of drug administration. To verify the accuracy of the design, a robust rodent ischemia model using the Middle Cerebral Arterial Occlusion (MCAO) technique was established. The effect of multiple combinations such as erythropoeitin derivatives, brain-derived neurotrophic factor and insulin-like growth factor (IGF-1) versus control is checked. MRI & Diffusion-weighted imaging is done to ensure similar ischemic lesions across patients and also reduction of hypoxic volume post-therapy. Behavioral monitoring using a battery of sensory-motor tests is done to correlate with biochemical and cellular changes. Results: On analysis of the effect of applying the computed dose of therapeutic agent at an optimal time point, on neural progenitor dynamics, we observed a strong peak of synaptic recovery. Findings based on animal experiments, MRI and histopathology provide empirical corroboration, thus establishing this approach to be useful for optimizing recovery in ischemic stroke. Conclusions: Given that the ischemic brain has evolved an incisive way to partly recoup itself by increasing the production of endogenous stem cell niches, the proposed approach can enable maximal/optimal recovery. Our efforts can be seen as the 1st endeavour of incorporating endogenous stem-cell processing influenced by neuro-modulators as a robust neuroinformatics template that allows for incorporation of patient specific parameters, thereby enabling one to optimize recovery using image-guided drug-scheduling. Keywords: computational neuroscience Conference: 5th INCF Congress of Neuroinformatics, Munich, Germany, 10 Sep - 12 Sep, 2012. Presentation Type: Poster Topic: Neuroinformatics Citation: Kapoor S, Rallabandi V and Roy P (2014). Optimizing Performance of Endogenous Neural Stem Cell Therapy for Ischemic Stroke: A Neuroinformatics and Neuroimaging Approach to Translational Medicine. Front. Neuroinform. Conference Abstract: 5th INCF Congress of Neuroinformatics. doi: 10.3389/conf.fninf.2014.08.00126 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 21 Mar 2013; Published Online: 27 Feb 2014. * Correspondence: Dr. Suhela Kapoor, National Brain Research Centre, Haryana, India, suhela.kapoor@nbrc.ac.in Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Suhela Kapoor VPS Rallabandi Prasun Roy Google Suhela Kapoor VPS Rallabandi Prasun Roy Google Scholar Suhela Kapoor VPS Rallabandi Prasun Roy PubMed Suhela Kapoor VPS Rallabandi Prasun Roy Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Pharmacokinetics of Piperacillin and Ciprofloxacin in Critically Ill Patients Undergoing Continuous Venovenous Haemodialysis or Haemodiafiltration

In sepsis an early time point of administration, adequate choice of an antibiotic drug and correct dosage are crucial for survival. Acute renal failure in severe sepsis can be treated by continuous renal replacement therapy but interferes with the pharmacokinetics of antibiotic drugs. The aim of the present study was to investigate the efficacy and safety of an antibiotic therapy with piperacillin/tazobactam and ciprofloxacin. In a single-center, prospective, open-label study a total of 24 patients with acute renal failure treated with continuous venovenous haemodialysis (CVVHD) or haemodiafiltration (CVVHDF) were enrolled in a clinical trial. Serum concentrations (Cmax, Cmin) and pharmacokinetic parameters of piperacillin and ciprofloxacin were analyzed. Optimum exposure to piperacillin is expected when serum concentrations are maintained 4-5 times higher than the minimum inhibitory concentration (MIC), i.e. above 64 mg/l. Optimum exposure to ciprofloxacin is given when the ratio (AUIC) of AUC and MIC is ≥ 125 h per dosing interval. In addition the Cmax/MIC ratio should amount to ≥ 10. Plasma concentrations lower than 64 mg/l were determined in 10 out of 21 patients treated with piperacillin. Nine out of 20 patients treated with ciprofloxacin had a calculated AUIC ≥ 125 h and a Cmax/MIC ratio ≥ 10. In critically ill patients undergoing CVVD or CVVDF piperacillin/tazobactam dosing should be increased to 4/0.5 g four times daily and ciprofloxacin dosing to 400 mg twice daily. Therapeutic drug monitoring of antibiotic therapies would be reasonable in these patients. The trial is registered at clinicaltrialsregister.eu ID: 2010-021369-66.

Topoisomerase I Inhibitors, Shikonin and Topotecan, Inhibit Growth and Induce Apoptosis of Glioma Cells and Glioma Stem Cells

Gliomas, the most malignant form of brain tumors, contain a small subpopulation of glioma stem cells (GSCs) that are implicated in therapeutic resistance and tumor recurrence. Topoisomerase I inhibitors, shikonin and topotecan, play a crucial role in anti-cancer therapies. After isolated and identified the GSCs from glioma cells successfully, U251, U87, GSCs-U251 and GSCs-U87 cells were administrated with various concentrations of shikonin or topotecan at different time points to seek for the optimal administration concentration and time point. The cell viability, cell cycle and apoptosis were detected using cell counting kit-8 and flow cytometer to observe the inhibitory effects on glioma cells and GSCs. We demonstrated that shikonin and topotecan obviously inhibited proliferation of not only human glioma cells but also GSCs in a dose- and time-dependent manner. According to the IC50 values at 24 h, 2 μmol/L of shikonin and 3 μmol/L of topotecan were selected as the optimal administration concentration. In addition, shikonin and topotecan induced cell cycle arrest in G0/G1 and S phases and promoted apoptosis. The down-regulation of Bcl-2 expression with the activation of caspase 9/3-dependent pathway was involved in the apoptosis process. Therefore, the above results showed that topoisomerase I inhibitors, shikonin and topotecan, inhibited growth and induced apoptosis of GSCs as well as glioma cells, which suggested that they might be the potential anticancer agents targeting gliomas to provide a novel therapeutic strategy.

Emerging cell and cytokine targets in rheumatoid arthritis

Despite major progress in the treatment of rheumatoid arthritis (RA), strong unmet medical need remains, as only a minor proportion of patients reach sustained clinical remission. New approaches are therefore necessary, and include manipulation of regulatory T cells, which might be able to restore the disturbed immune system and could even lead to a cure if this restored regulation were to prove sustainable. Logistical and conceptual problems, however, beset this attractive therapeutic approach, including difficulties with ex vivo expansion of cells, specificity of targeting and the optimal time point of administration. Therefore, alternative avenues are being investigated, such as targeting B-cell effector functions and newly identified proinflammatory cytokines. On the basis of success with B-cell depleting therapy using anti-CD20 agents, further treatment modalities are now exploring direct or indirect interference in B-cell-mediated immunity with the use of agents directed against other B-cell surface molecules. Novel approaches target intracellular B-cell signalling and regulatory B cells. New cytokine-directed therapies target important proinflammatory mediators such as GM-CSF, new members of the IL-1 family, IL-6 and its receptor, IL-17, IL-20, IL-21, IL-23 as well as synovium-specific targets. This article reviews these emerging cell and cytokine targets with special focus on biologic agents, some of which might reach the clinic soon whereas others will require considerable time in development. Nevertheless, these exciting new approaches will considerably enhance our repertoire in the battle against this potentially devastating disease.

The Utility of the KJOC Score in Professional Baseball in the United States

Background: The Kerlan-Jobe Orthopaedic Clinic (KJOC) Shoulder and Elbow questionnaire has been shown by previous studies to be more sensitive than other validated subjective measurement tools in the detection of upper extremity dysfunction in overhead-throwing athletes. Purpose: The primary objective was to establish normative data for KJOC scores in professional baseball players in the United States. The secondary objectives were to evaluate the effect of player age, playing position, professional competition level, history of injury, history of surgery, and time point of administration on the KJOC score. Study Design: Cross-sectional study; Level of evidence, 3. Methods: From 2011 to 2012, a total of 203 major league and minor league baseball players within the Houston Astros professional baseball organization completed the KJOC questionnaire. The questionnaire was administered at 3 time points: spring training 2011, end of season 2011, and spring training 2012. The KJOC scores were analyzed for significant differences based on player age, injury history, surgery history, fielding position, competition level, self-reported playing status, and time point of KJOC administration. Results: The average KJOC score among healthy players with no history of injury was 97.1 for major league players and 96.8 for minor league players. The time point of administration did not significantly affect the final KJOC score (P = .224), and KJOC outcomes did not vary with player age (r = −0.012; P = .867). Significantly lower average KJOC scores were reported by players with a history of upper extremity injury (86.7; P < .001) and upper extremity surgery (75.4; P < .0001). The KJOC results did vary with playing position (P = .0313), with the lowest average scores being reported by pitchers (90.9) and infielders (91.3). Conclusion: This study establishes a quantitative baseline for the future evaluation of professional baseball players with the KJOC score. Age and time of administration had no significant effect on the outcome of the KJOC score. Missed practices or games within the previous year because of injury were the most significant demographic predictors of lower KJOC scores. The KJOC score was shown to be a sensitive measurement tool for detecting subtle changes in the upper extremity performance of the professional baseball population studied.

Pre- and postconditioning effect of Sevoflurane on myocardial dysfunction after cardiopulmonary resuscitation in rats

Post-resuscitation myocardial dysfunction is an important cause of death in the intensive care unit after initially successful cardiopulmonary resuscitation (CPR) of pre-hospital cardiac arrest (CA) patients. Volatile anaesthetics reduce ischaemic–reperfusion injury in regional ischaemia in beating hearts. This effect, called anaesthetic-induced pre- or postconditioning, can be shown when the volatile anaesthetic is given either before regional ischaemia or in the reperfusion phase. However, up to now, little data exist for volatile anaesthetics after global ischaemia due to CA. Therefore, the goal of this study was to clarify whether Sevoflurane improves post-resuscitation myocardial dysfunction after CA in rats.Following institutional approval by the Governmental Animal Care Committee, 144 male Wistar rats (341±19g) were randomized either to a control group or to one of the 9 interventional groups receiving 0.25 MAC, 0.5 MAC or 1 MAC of Sevoflurane for 5min either before resuscitation (SBR), during resuscitation (SDR) or after resuscitation (SAR). After 6min of electrically induced ventricular fibrillation CPR was performed. Before CA (baseline) as well as 1h and 24h after restoration of spontaneous circulation (ROSC), continuous measurement of ejection fraction (EF), and preload adjusted maximum power (PAMP) as primary outcome parameters and end systolic pressure (ESP), end diastolic volume (EDV) and maximal slope of systolic pressure increment (dP/dtmax) as secondary outcome parameters was performed using a conductance catheter.EF was improved in all Sevoflurane treated groups 1h after ROSC in comparison to control, except for the 0.25 MAC SDR and 0.25 MAC SAR group (0.25 MAC SBR: 38±8, p=0.02; 0.5 MAC SBR: 39±7, p=0.04; 1 MAC SBR: 40±6, p=0.007; 0.5 MAC SDR: 38±7, p=0.02; 1 MAC SDR: 40±6, p=0.006; 0.5 MAC SAR: 39±6, p=0.01; 1 MAC SAR: 39±6, p=0.002, vs. 30±7%). Twenty-four hours after ROSC, EF was higher than control in all interventional groups (p<0.05 for all groups). EF recovered to baseline values 24h after ROSC in all SBR and SAR groups. PAMP was improved in comparison to control (4.6±3.0mW/μl2) 24h after ROSC in 0.5 MAC SBR (9.4±6.9mW/μl2, p=0.04), 1 MAC SBR (8.9±4.4mW/μl2, p=0.04), 1 MAC SDR (8.0±5.7mW/μl2, p=0.04), and 1 MAC SAR (7.3±3.5mW/μl2, p=0.04). ESP, EDV, and dP/dtmax was not different from control 1h as well as 24h after ROSC with the exception of 1 MAC SDR with a reduced ESP 1h after ROSC (89±16 vs. 103±22mmHg, p=0.04). Sevoflurane treatment did not affect survival rate.This animal study of CA and resuscitation provides the hypothesis that pharmacological pre- or postconditioning with the volatile anaesthetic Sevoflurane – administered before CA, during resuscitation or after ROSC – results in an improved myocardial inotropy 24h after ROSC. Sevoflurane treatment seems to improve EF even in the early phase of reperfusion 1h after ROSC. Therefore further targeted studies on the optimal dose and time point of administration of Sevoflurane in cardiopulmonary resuscitation seem to be worthwhile (Institutional protocol number: 35-9185.81/G-24/08).