Search

Background: Cannabis (CAN) use has risen significantly over the last few decades. CAN has potent immunosuppressive properties, which could antagonize the effect of immunotherapy (IO). The impact of CAN use on clinical cancer outcomes remains unclear., Objectives: In this study, we evaluated the clinical effect of CAN use on clinical outcomes among patients with solid malignancies receiving IO., Design: This is a retrospective cohort study of all patients with solid malignancies receiving IO between August 2014 and August 2018., Methods: Patients were stratified based on CAN use to CAN users and CAN non-users. The primary outcome was overall survival (OS), and the secondary outcomes were progression-free survival (PFS) and disease control rate (DCR). Univariable and multivariable logistic and Cox regression analyses were performed to compare the outcomes between the two groups, adjusting for covariates., Results: The records of 106 patients were reviewed, 28 (26%) of whom were CAN users and 78 (74%) were CAN non-users. One patient was excluded. Most CAN users consumed dronabinol (82%). The median follow-up for OS and PFS was 29.2 months. Median OS in the CAN users was 6.7 months compared to 17.3 months in the CAN non-users (HR, 1.78; 95% CI, 1.06-2.97; p  = 0.029). The median PFS was 4.8 months in the CAN users compared to 9.7 months in the CAN non-users (HR, 1.74; 95% CI, 1.09-2.79; p  = 0.021). DCR was 11% among CAN users and 38% among CAN non-users (OR, 0.23; 95% CI; 0.06-0.68; p  = 0.007). An exploratory racial disparity analysis showed that this negative impact of CAN was primarily seen in White patients., Conclusion: In this single institutional experience, CAN use was associated with worse OS, PFS, and DCR among cancer patients receiving IO. Prospective trials are needed to further study this potential antagonistic interaction between CAN and IO and explore the racial disparities related to CAN exposure., (© The Author(s), 2024.)

Inotuzumab ozogamicin (IO), a novel therapeutic drug for relapsed or refractory acute lymphoblastic leukemia (RR)‑(ALL), is a humanized anti‑cluster of differentiation (CD) 22 monoclonal antibody conjugated with calicheamicin that causes DNA single‑ and double‑strand breaks. Although the efficacy of IO is significantly improved compared with that of conventional chemotherapies, the prognosis for RR‑ALL remains poor, highlighting the need for more effective treatment strategies. The present study examined the role of DNA damage repair inhibition using the poly (ADP‑ribose) polymerase (PARP) inhibitors olaparib or talazoparib on the enhancement of the antitumor effects of IO on B‑ALL cells in vitro . The Reh, Philadelphia (Ph) ‑ B‑ALL and the SUP‑B15 Ph + B‑ALL cell lines were used for experiments. Both cell lines were ~90% CD22 + . The half‑maximal inhibitory concentration (IC 50 ) values of IO were 5.3 and 49.7 ng/ml for Reh and SUP‑B15 cells, respectively. The IC 50 values of IO combined with minimally toxic concentrations of olaparib or talazoparib were 0.8 and 2.9 ng/ml for Reh cells, respectively, and 36.1 and 39.6 ng/ml for SUP‑B15 cells, respectively. The combination index of IO with olaparib and talazoparib were 0.19 and 0.56 for Reh cells and 0.76 and 0.89 for SUP‑B15 cells, demonstrating synergistic effects in all combinations. Moreover, the addition of minimally toxic concentrations of PARP inhibitors augmented IO‑induced apoptosis. The alkaline comet assay, which quantitates the amount of DNA strand breaks, was used to investigate the degree to which DNA damage observed 1 h after IO administration was repaired 6 h later, reflecting successful repair of DNA strand breaks. However, DNA strand breaks persisted 6 h after IO administration combined with olaparib or talazoparib, suggesting inhibition of the repair processes by PARP inhibitors. Adding olaparib or talazoparib thus synergized the antitumor effects of IO by inhibiting DNA strand break repair via the inhibition of PARP.

Background: Understanding how to modulate the microenvironment of tumors that are resistant to immune checkpoint inhibitors represents a major challenge in oncology.Here we investigate the ability of USP7 inhibitors to reprogram the tumor microenvironment (TME) by inhibiting secretion of vascular endothelial growth factor (VEGF) from fibroblasts., Methods: To understand the role played by USP7 in the TME, we systematically evaluated the effects of potent, selective USP7 inhibitors on co-cultures comprising components of the TME, using human primary cells. We also evaluated the effects of USP7 inhibition on tumor growth inhibition in syngeneic models when dosed in combination with immune checkpoint inhibitors (ICIs)., Results: Abrogation of VEGF secretion from fibroblasts in response to USP7 inhibition resulted in inhibition of tumor neoangiogenesis and increased tumor recruitment of CD8-positive T-lymphocytes, leading to significantly improved sensitivity to immune checkpoint inhibitors. In syngeneic models, treatment with USP7 inhibitors led to striking tumor responses resulting in significantly improved survival., Conclusions: USP7-mediated reprograming of the TME is not linked to its previously characterized role in modulating MDM2 but does require p53 and UHRF1 in addition to the well-characterized VEGF transcription factor, HIF-1α. This represents a function of USP7 that is unique to fibroblasts, and which is not observed in cancer cells or other components of the TME. Given the potential for USP7 inhibitors to transform "immune desert" tumors into "immune responsive" tumors, this paves the way for a novel therapeutic strategy combining USP7 inhibitors with immune checkpoint inhibitors (ICIs)., (© 2024 Almac Discovery Ltd. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.)

A CCSD(T)//B3LYP method was employed to research the influence of monomolecular and bimolecular water molecules on the IO + CH 2 O reaction. H-abstraction and addition/elimination mechanisms have been located, and the H-abstraction mechanism occupied the whole reaction without water. The introduction of water complicates the reaction, but the final product remains unchanged. Water molecules influence each reaction pathway in six different pathways with monomolecular water and two different pathways with bimolecular water, respectively. The present calculations manifested that water has a positive influence on the IO + CH 2 O reaction with monomolecular water molecule, and plays negative catalytic influence on the IO + CH 2 O reaction with bimolecular water molecule. The effective rate constants are much lower than the IO + CH 2 O reaction without water., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Working with patients who end their lives through suicide is one of the greatest challenges and fears a clinician might face. This paper explores the experience of working with such patients in the course of psychiatric care and Jungian analysis. Jungian theory is used to explore a conceptual understanding of the dilemmas faced. Such theory points to an intractable conflict between Self and ego giving rise to unbearable distress, partly through difficulties arising in achieving individuation. The paper proposes that our difficulties in dealing with a patient's suicide might reflect a manifestation of relentless hope and, perhaps contentiously, these patients made a more considered decision in their final acts than prevailing clinical opinion suggests., (© 2022, The Society of Analytical Psychology.)

Intraosseous infusion (IO) remains an underutilized technique for obtaining vascular access in adults, despite its potentially life-saving benefits in trauma patients. In rural and remote areas, shortage of training equipment and human capacity (i.e., simulators) are the main contributors to the shortage of local training courses aiming at the development and maintenance of IO skills. Specifically, current training equipment options available for trainees include commercially available simulators, which are often expensive, or animal tissues, which lack human anatomical features that are necessary for optimal learning and pose logistical and ethical issues related to practice on live animals. Three-dimensional (3D) printing provides the means to create cost-effective, anatomically correct simulators for practicing IO where existing simulators may be difficult to access, especially in remote areas. This technical report aims to describe the development of maxSIMIO, a 3D-printed adult proximal tibia IO simulator, and present feedback on the design features from a clinical co-design team consisting of 18 end-point users.  Overall, the majority of the feedback was positive and highlighted that the maxSIMIO simulator was helpful for learning and developing the IO technique. The majority of the clinical team responders also agreed that the simulator was more anatomically accurate compared to other simulators they have used in the past. Finally, the survey results indicated that on average, the simulator is acceptable as a training tool. Notable suggestions for improvement included increasing the stability of the individual parts of the model (such as tightening the skin and securing the bones), enhancing the anatomical accuracy of the experience (such as adding a fibula), making the bones harder, increasing the size of the patella, making it more modular (to minimize costs related to maintenance), and improving the anatomical positioning of the knee joint (i.e., slightly bent in the knee joint). In summary, the clinical team, located in rural and remote areas in Canada, found the 3D-printed simulator to be a functional tool for practicing the intraosseous technique. The outcome of this report supports the use of this cost-effective simulator for simulation-based medical education for remote and rural areas anywhere in the world., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Sivanathan et al.)

Background: The efficacy of tyrosine kinase inhibitor (TKI)-based therapy after previous immuno-oncology therapy (IO) failure has been addressed before. However, summary efficacy estimates have never been generated in these reports. We addressed this void., Material and Methods: We systematically examined TKI efficacy after IO-failure and generated weighted median progression-free survival (PFS) estimates for Pazopanib, Axitinib, Cabozantinib, Sunitinib. A systematic review according to PRISMA was conducted. PubMed and abstracts were queried. Only studies proving median PFS were included. Weighted medians were computed for each TKI alternative., Results: Of 245 articles, nine eligible studies were included in the current study with 952 analysed patients. Weighted PFS medians after any previous IO-based therapy were respectively 13.7 (range from 4.6 to 24.4), 8.1 (range from 4.7 to 13.2), 8.5 (range from 4.7 to 15.2) and 6.9 months (range from 2.9 to 11.6) for Pazopanib, Axitinib, Cabozantinib, Sunitinib. Specific second-line weighted PFS median was 14.8 months (range from 5.6 to 24.4), 10.1 months (range from 6.4 to 13.2), 8.7 months (range from 4.7 to 15.2) and 6.0 months (range from 2.9 to 8.0) for Pazopanib, Axitinib, Cabozantinib, Sunitinib, respectively, after first-line IO., Conclusion: Pazopanib results in the longest weighted median PFS, after previous IO-failure, regardless of treatment line, as well as in specific second-line, post-first-line IO failure settings. Pending novel studies, Pazopanib appears to represent the most promising treatment option after prior IO., Competing Interests: Conflict of interest statement The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Aim: Intraosseous (IO)-access plays an alternative route during resuscitation. Our study in preterm and term stillborns was performed to find alternative IO puncture sites beside the recommended proximal tibia., Methods: The cadavers used were legal donations. 20 stillborns (mean: 29.2weeks, IQR 27.1-39.6) were investigated. Spectral-CT were analysed to calculate the diameter and circumferences of: i) proximal humerus ii) distal femur iii) proximal tibia iv) diaphyseal tibial. Contrast medium was applied under video documentation to investigate the drainage into the vascular system., Results: In term newborns, diameter of the cortex of the proximal humeral head is 12.1 ± 1.8 mm, distal end of the femur 11.9 ± 3.4 mm and the proximal tibial bone 12.0 ± 2.4 mm with cross-sectional diameter of 113.5 ± 19.7 mm 2 , 120.6 ± 28.2 mm 2 and 111.6 ± 29.5 mm 2 , respectively. Regarding the preterm groups, there is a strong age-related growth in diameter and cross -sectional size. The diaphyseal area is the smallest in all measured bones with an age-dependent increase and is about half of that of metaphyseal diameters (proximal and distal) and about one third of that of metaphyseal cross sectional areas. The proximal femoral head region has the largest diameter of all measured bones with an egg-shaped formation with an extensive joint capsula. All investigated metaphyseal areas lack a clearly enclosed bone marrow cavity. Infusion of contrast medium into the distal femoral end and the proximal humerus head demonstrate the drainage of contrast medium into the central venous system within seconds., Conclusion: Proximal humeral head and distal femoral end might be alternative IO areas which may lead to further IO puncture sites in neonates., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

The traditional treatments for esophageal squamous cell carcinoma include surgery and radiation as local therapies, then chemotherapy and targeted therapy as systemic treatments. These treatments, either alone or in combination, however, are not satisfactory because of limited efficacy and unfavorable toxicity, calling for new therapeutic strategies. In recent years, immunotherapy, a new weapon in the arsenal against cancer, has shown substantial clinical benefits in patients with esophageal squamous cell carcinoma, particularly ones with locally advanced or metastatic disease. Importantly, accumulating evidence suggests that traditional radiation therapy functions as a powerful adjuvant for immunotherapy by contributing to systemic antitumor immunity, resulting in reduced recurrence risk and improved survival of patients. Here the authors summarize the emerging data on immunotherapy- and radiation therapy-based treatment of esophageal squamous cell carcinoma and discuss the pros and cons of different combinations, aiming at a comprehensive understanding of the proper rationale for the design of effective therapeutic regimens.

Background: Second-line immunotherapy (IO) has shown an overall survival benefit. However, only 18% to 20% of patients with advanced non-small-cell lung cancer (aNSCLC) will respond, with a median progression-free survival (PFS) of 2 to 4 months. Thus, biomarkers to select those patients most likely to benefit from IO are greatly needed., Patients and Methods: We conducted a retrospective analysis of 154 patients with aNSCLC who had received anti-programmed cell death 1 therapy as second line or further treatment. We assessed the absolute neutrophil, lymphocyte, monocyte, and eosinophil counts at baseline (T0) and the second (T1) and third (T2) cycles. The neutrophil/lymphocyte ratio (NLR), derived-NLR (dNLR), lymphocyte/monocyte ratio (LMR), and their percentage of change at T1 and T2 compared with T0 were evaluated. The clinical characteristics and lactate dehydrogenase (LDH) level were also considered. Univariate and multivariate analyses were performed. Significant biomarkers for PFS on multivariate analysis were combined in a prognostic score., Results: For overall survival, the negative prognostic biomarkers were Eastern Cooperative Oncology Group (ECOG) performance status (PS) 2, NLR at T0, and dNLR at T1; the LMR at T0, T1, and T2 was identified as a positive prognostic biomarker. For PFS, the negative prognostic biomarkers were ECOG PS 2, liver metastases, NLR at T0, dNLR at T1 and T2, and ≥ 30% increase of NLR from T0 to T1; the positive prognostic biomarkers were heavy smoking, LDH, and LMR at T2. The ≥ 30% increase of LMR from T0 to T1 and T0 to T2 correlated with the overall response rate. A prognostic score (EPSILoN score; smoking, ECOG PS, liver metastases, LDH, NLR) identified 3 prognostic groups (median PFS, 10.2, 4.9, and 1.7 months, respectively; P < .001)., Conclusions: The EPSILoN score combines 5 baseline clinical and blood biomarkers and can help to identify patients with aNSCLC who will most likely benefit from second-line IO. Further studies are warranted., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The Jovian polar regions produce X-rays that are characteristic of very energetic oxygen and sulfur that become highly charged on precipitating into Jupiter's upper atmosphere. Juno has traversed the polar regions above where these energetic ions are expected to be precipitating revealing a complex composition and energy structure. Energetic ions are likely to drive the characteristic X-rays observed at Jupiter (Haggerty et al., 2017, https://doi.org/10.1002/2017GL072866; Houston et al., 2018, https://doi.org/10.1002/2017JA024872; Kharchenko et al., 2006, https://doi.org/10.1029/2006GL026039). Motivated by the science of X-ray generation, we describe here Juno Jupiter Energetic Particle Detector Instrument (JEDI) measurements of ions above 1 MeV and demonstrate the capability of measuring oxygen and sulfur ions with energies up to 100 MeV. We detail the process of retrieving ion fluxes from pulse width data on instruments like JEDI (called "puck's"; Clark, Cohen, et al., 2016, https://doi.org/10.1002/2017GL074366; Clark, Mauk, et al., 2016, https://doi.org/10.1002/2015JA022257; Mauk et al., 2013, https://doi.org/10.1007/s11214-013-0025-3) as well as details on retrieving very energetic particles (>20 MeV) above which the pulse width also saturates., (©2019. The Authors.)

The following article elaborates on the need for the Other in order to delineate the self and one's identity, and the eternal psychological process of turning the Other into an enemy. A parallel process, which can be observed nowadays, leads to a disregard for the otherness of the Other that results in the blurring of identity and an internal loss of the differentiation of intrapsychic structures. Both psychological processes share a dehumanizing approach which leads to neurosis and alienation. In order to withstand these tendencies, a revision of analytical psychology's view of the psyche and of the practice of Jungian analysis is warranted: an alternative to the 'Fear of the Feminine' described by Erich Neumann in 1959. It is suggested that consciousness and unconsciousness intertwine but from a position of equality and reciprocity. Such an attitude does not inspire fear. Rather, it recognizes the need of one for the Other and the inevitability of this situation. Moreover, this need and interdependence on equal grounds nourish the wish to know the Other, to be aware of the differences, and yet, at the same time, to acknowledge how close the Other really is. Analytical psychological therapy based on this model stresses the power of the ego while also strengthening its capacity for introspection., (© 2019, The Society of Analytical Psychology.)

Background: Programmed death-ligand 1 (PD-L1) expression in metastatic renal cell carcinoma (RCC) correlates with a worse prognosis, but whether it also predicts responsiveness to anti-PD-1/PD-L1 therapy remains unclear. Most studies of PD-L1 are limited by evaluation in primary rather than metastatic sites, and in biopsy samples, which may not be representative. These limitations may be overcome with immuno-positron emission tomography (iPET), an emerging tool allowing the detection of cell surface proteins with radiolabeled antibodies. Here, we report iPET studies of PD-L1 in a preclinical tumorgraft model of clear cell RCC (ccRCC) from a patient who had a favorable response to anti-PD-1 therapy., Case Presentation: A 49-year-old man underwent a cytoreductive nephrectomy in 2017 of a right kidney tumor invading into the adrenal gland that was metastatic to the lungs and a rib. Histological analyses revealed a ccRCC of ISUP grade 4 with extensive sarcomatoid features. IMDC risk group was poor. Within two hours of surgery, a tumor sample was implanted orthotopically into NOD/SCID mice. Consistent with an aggressive tumor, a renal mass was detected 18 days post-implantation. Histologically, the tumorgraft showed sarcomatoid differentiation and high levels of PD-L1, similar to the patient's tumor. PD-L1 was evaluated in subsequently transplanted mice using iPET and the results were compared to control mice implanted with a PD-L1-negative tumor. We labeled atezolizumab, an anti-PD-L1 antibody with a mutant Fc, with zirconium-89. iPET revealed significantly higher 89 Zr-atezolizumab uptake in index than control tumorgrafts. The patient was treated with high-dose IL2 initially, and subsequently with pazopanib, with rapidly progressive disease, but had a durable response with nivolumab., Conclusions: To our knowledge, this is the first report of non-invasive detection of PD-L1 in renal cancer using molecular imaging. This study supports clinical evaluation of iPET to identify RCC patients with tumors deploying the PD-L1 checkpoint pathway who may be most likely to benefit from PD-1/PD-L1 disrupting drugs.

A conceptual design is presented for a low complexity, heritage-based flyby mission to Io, Jupiter's innermost Galilean satellite and the most volcanically active body in the Solar System. The design addresses the 2011 Decadal Surveys recommendation for a New Frontiers class mission to Io and is based upon the result of the June 2012 NASA-JPL Planetary Science Summer School. A science payload is proposed to investigate the link between the structure of Io's interior, it's volcanic activity, it's surface composition, and it's tectonics. A study of Io's atmospheric processes and Io's role in the Jovian magnetosphere is also planned. The instrument suite includes a visible/near IR imager, a magnetic field and plasma suite, a dust analyzer and a gimbaled high gain antenna to perform radio science investigations. Payload activity and spacecraft operations would be powered by three Advanced Stirling Radioisotope Generators (ASRG). The primary mission includes 10 flybys with close-encounter altitudes as low as 100 km. The mission risks are mitigated by ensuring that relevant components are radiation tolerant and by using redundancy and flight-proven parts in the design. The spacecraft would be launched on an Atlas V rocket with a delta- v of 1.3 km/s. Three gravity assists (Venus, Earth, Earth) would be used to reach the Jupiter system in a 6-year cruise. The resulting concept demonstrates the rich scientific return of a flyby mission to Io.

Objective: To compare the time required and the success rate of personnel with 4 different levels of experience to place a humeral intraosseous (IO) catheter versus a jugular venous catheter (IV) in cadaver dogs., Design: Prospective study., Setting: Veterinary university teaching hospital., Interventions: Canine cadavers from recently euthanized dogs were obtained from the cadaver donation program between May and December 2014. Catheter placers (CPs) with varying clinical experience, including a first year emergency and critical care resident, a senior emergency veterinary technician (VTS certified), a final year veterinary student, and an ACVECC diplomate, participated in the study. Each CP catheterized a total of 6 dogs so that there was a total of 6 IO and 6 IV catheters placed, by automatic rotary insertion device (with an EZ-IO gun) and vascular cut-down technique, respectively, for each CP. Time for IO catheterization and IV catheterization was recorded and compared. The success of IO catheterization and IV catheterization was verified by visualization of an injection of iodinated contrast material under fluoroscopy within the medullary cavity or vessel., Animals: Twenty-four canine cadavers., Measurements and Main Results: Outcomes were analyzed using the Wilcoxon rank-sum test and the Kruskal-Wallis one-way analysis of variance. The median time for all IO catheterization operators was faster at 55.4 seconds (range 15.0-153.0 s) compared to the median time for all IV catherization operators at 217.3 seconds (range 55.6-614 s). The success rate for IO and IV was equal at 87.5%., Conclusion: IO catheterization using an automatic rotary insertion device was performed more rapidly and successfully than jugular venous catheterization using a cut-down technique in canine cadaver. These findings suggest IO catheterization may be more efficient for gaining vascular access in the appropriate emergency clinical situations when preexisting IV access does not exist., (© Veterinary Emergency and Critical Care Society 2017.)

Background & Aims: We investigated whether tissue elastography (TE) can be used as an alternative to magnetic resonance imaging (MRI) T2* analysis to determine the degree of iron overload in patients with thalassemia major., Methods: We conducted a prospective study of 154 patients (99 male; mean age, 12 ± 3.6 years) with thalassemia major requiring chronic blood transfusion and on iron chelator therapy. The study was performed at a tertiary hospital in India from January 2015 through June 2015. We performed routine blood sample analyses, measurements of serum levels of ferritin, and TE within 1 month of MRI T2* analysis of the liver. The Spearman correlation test and linear regression analysis were used to evaluate the correlation between TE liver stiffness measurements and R2* MRI results or serum ferritin levels., Results: The subjects' mean total serum levels of bilirubin, alanine aminotransferase, aspartate aminotransferase, and albumin were 1.4 ± 0.6 mg/dL, 65.0 ± 51.8 IU/L, 62.9 ± 44 IU/L, and 4.2 ± 0.2 g/d, respectively. Mean liver stiffness measurement, MRI T2* (3 T), corresponding MRI R2* (3 T), and ferritin values were 8.2 ± 4.4 kPa, 3.18 ± 2.6 milliseconds, 617.3 ± 549 Hz, and 4712 ± 3301 ng/mL, respectively. On the basis of MRI analysis, 67 patients (43.5%) had mild iron overload, 49 patients (31.8%) had moderate iron overload, and 22 patients (14.3%) had severe iron overload. Fibroscan liver stiffness measurements correlated with MRI R2* values (r = 0.85; P < .001). TE results identified the patients with severe, moderate, and mild iron overload with area under the receiver operating characteristic curve values of 94.8%, 84.5%, and 84.7%, respectively. Liver stiffness measurements greater than 13.5, 7.8, and 5.5 kPa identified patients with severe, moderate, and mild iron overload, respectively; the sensitivity and specificity values were 92% and 93% for severe overload, 82% and 82% for moderate overload, and 73% and 75% for mild overload. No correlation was found between TE results and serum level of ferritin (r = 0.19; P = .11)., Conclusions: Results of TE correlate with those from MRI T2* analysis. TE is cheaper and more available than MRI and might be used to estimate hepatic iron overload, especially moderate to severe overload in patients with thalassemia major who require chronic transfusion., (Copyright © 2017 AGA Institute. Published by Elsevier Inc. All rights reserved.)

Introduction: There is a lack of information regarding intraosseous (IO) administration of tranexamic acid (TXA). Our hypothesis was that a single bolus IO injection of TXA will have a similar pharmacokinetic profile to TXA administered at the same dose IV., Methods: Sixteen male Landrace cross swine (mean body weight 27.6±2.6kg) were divided into an IV group (n=8) and an IO group (n=8). Each animal received 30mg/kg TXA via an IV or IO catheter, respectively. Jugular blood samples were collected for pharmacokinetic analysis over a 3h period. The maximum TXA plasma concentration (C max ) and corresponding time as well as distribution half-life, elimination half-life, area under the curve, plasma clearance and volume of distribution were calculated. One- and two-way analysis of variance for repeated measures (time, group) with Tukey's and Bonferonni post hoc tests were used to compare TXA plasma concentrations within and between groups, respectively., Results: Plasma concentrations of TXA were significantly higher (p<0.0001) in the IV group during the TXA infusion. C max occurred at 4min after initiation of the bolus in the IV group (9.36±3.20ng/μl) and at 5min after initiation of the bolus in the IO group (4.46±0.49ng/μl). Plasma concentrations were very similar from the completion of injection onwards. There were no significant differences between the two administration routes for any other pharmacokinetic variables measured., Conclusion: The results of this study support pharmacokinetic bioequivalence of IO and IV administration of TXA., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Pompe disease is a clinically and genetically heterogeneous autosomal recessive disorder caused by lysosomal acid α-glucosidase (GAA) deficiency. We report on two affected members of a non-consanguineous Caucasian family, including a classical infantile-onset patient with severe cardiomyopathy (IO) and his paternal grandmother with the adult-onset (AO) form. Two compound heterozygous sequence variants of the GAA gene were identified in each patient by mutation analyses (IO=c.1211A>G and c.1798C>T; AO=c.1211A>G and c.692+5G>T). For this study, the biochemical phenotype resulting from the missense mutation c.1211A>G in exon 8, which converts a highly conserved aspartate to glycine (p.Asp404Gly), was of specific interest because it had not been reported previously. Western blotting revealed a robust expression of all GAA isoforms in quadriceps muscle of both patients (fully CRIM positive), while enzymatic activity was 3.6% (IO) and 6.6% (AO) of normal controls. To further validate these findings, the c.1211A>G sequence variant was introduced in wild type GAA cDNA and over-expressed in HEK293T cells. Site-directed mutagenesis analyses confirmed that the mutation does not affect processing or expression of GAA protein, but rather impairs enzyme function. Similar results were reported for c.1798C>T (p.Arg600Cys), which further supports the biochemical phenotype observed in IO. The third mutation (c.692+5G>T, in intron 3) was predicted to affect normal splicing of the GAA mRNA, and qPCR indeed verified a 4-fold lower mRNA expression in AO. It is concluded that the novel sequence variant c.1211A>G results in full CRIM but significantly lower GAA activity, which in combination with c.1798C>T leads to infantile-onset Pompe disease. We surmise that the difference in disease severity between the two family members in this study is due to a milder effect of the intronic mutation c.692+5G>T (vs. c.1798C>T) on phenotype, partially preserving GAA activity and delaying onset in the proband (paternal grandmother)., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Marine organisms have bioactive potential which has tremendous pharmaceutical promise. Emerging evidence highlights the importance of the interplay between bone and the immune system of which T lymphocytes and their product act as key regulators of bone resorption. In the present investigation we have analyzed the anti-osteoporotic effect of turbo methanol extract (TME) in the reversal of bone resoprtion. Forty-two female Swiss albino mice were used and randomly assigned into sham-operated group (sham) and six ovariectomized (OVX) subgroups, i.e. OVX with vehicle (OVX) that received daily oral administration of water ad libitum; OVX with estradiol (2mg/kg/day); and OVX with different doses of TME i.e. TME 100mg/kg, TME 50mg/kg, TME 25mg/kg and TME 12.5mg/kg. Oral administration of TME or estradiol started on the second week after ovariectomy for a period of 4weeks. We observed that the administration of TME increased the trabeculation in tibia and reduced the atrophy in the uterus. TME significantly decreased the serum alkaline phosphatase (ALP) and acid phosphatase (ACP) activity in OVX mice. Micro CT analysis revealed that the TME administration preserved the bone volume, connectivity density, trabecular number, trabecular thickness and trabecular separation in OVX mice. Bone mineralization was measured in different groups of mice by Raman spectroscopy. Reversal of bone resorption was observed in TME treated group of mice. To further investigate the mechanism of action of TME, we analyzed the T lymphocyte proliferation and profiles of cytokine TNFα and sRANKL in TME treated ovariectomized mice. Decrease in the elevation of T cell subsets was observed after the supplementation with TME. The extract significantly lowered the T cell proliferation responses to mitogens, phorbol 12-myristate 13-acetate (PMA) and ionomycin (Io) and phytohemagglutinin (PHA). A marked reduction in TNFα and sRANKL secretion in serum and TNFα in cell free supernatants of activated T lymphocytes was observed upon TME administration. TME could significantly inhibit the in vitro osteoclastogenesis and the bone resorption observed using artificial calcium coated slides. Collectively, these results indicate that TME has the potential to inhibit bone resorption and may prove to be a potential candidate for the development of an anti-osteoporosis drug., (© 2013.)

This study clarified the neural mechanisms underlying jaw movements in pharyngolaryngeal reflexes such as swallowing in rats. After retrograde tracer injections into the ventromedial division (Vmovm) of the trigeminal motor nucleus (Vmo) containing jaw-opening (JO) motoneurons or into the dorsolateral division (Vmodl) of Vmo containing jaw-closing (JC) motoneurons, JO and JC premotoneurons were labeled with an ipsilateral predominance in the medial and intermediate subnuclei of the rostrocaudal middle two-thirds of the nucleus of the solitary tract (Sol); JC premotoneurons were also in the lateral subnucleus of Sol. After anterograde tracer injections into the Sol, axons were labeled with an ipsilateral predominance in the Vmovm and Vmodl, prominently in the ipsilateral Vmovm. After transganglionic tracer applications to the superior laryngeal nerve (SLN) or the cervical trunk of the glossopharyngeal nerve (GpN-ct), labeled afferents were seen in the medial, intermediate, lateral and interstitial subnuclei of Sol at the rostral three-fourths of Sol, indicating considerable overlap with the JO and JC premotoneurons in the Sol. Double labeling experiments demonstrated contacts between the afferent terminals and the JO and JC premotoneurons. The present study has for the first time revealed the differential distribution of JO and JC premotoneurons in the Sol and features of their projections from the Sol, as well as their connections with SLN and GpN-ct afferent inputs. The JO and JC premotoneurons in the Sol may play an important role in generation and organization of jaw movements in pharyngolaryngeal reflexes evoked by SLN and GpN-ct inputs, such as swallowing., (© 2013 Published by Elsevier B.V.)

Gene electrotransfer (GET) enhances delivery of DNA vaccines by increasing both gene expression and immune responses. Our lab has developed the multi-electrode array (MEA) for DNA delivery to skin. The MEA was used at constant pulse duration (150 ms) and frequency (6.67 Hz). In this study, delivery parameters including applied voltage (5-45 V), amount of plasmid (100-300 μg), and number of treatments (2-3) were evaluated for delivery of a DNA vaccine. Mice were intradermally injected with plasmid expressing Bacillus anthracis protective antigen with or without GET and αPA serum titers measured. Within this experiment no significant differences were noted in antibody levels from varying dose or treatment number. However, significant differences were measured from applied voltages of 25 and 35 V. These voltages generated antibody levels between 20,000 and 25,000. Serum from animals vaccinated with these conditions also resulted in toxin neutralization in 40-60% of animals. Visual damage was noted at MEA conditions of 40 V. No damage was noted either visually or histologically from conditions of 35 V or below. These results reflect the importance of establishing appropriate electrical parameters and the potential for the MEA in non-invasive DNA vaccination against B. anthracis., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Cerebral ischemia is encompassed by cerebrovascular apoptosis, yet the mechanisms behind apoptosis regulation are not fully understood. We previously demonstrated inhibition of endothelial apoptosis by vascular endothelial growth factor (VEGF) through upregulation of poly(ADP-ribose)-polymerase (PARP) expression. However, PARP overactivation through oxidative stress can lead to necrosis. This study tested the hypothesis that neuropilin-1 (NP-1), an alternative VEGF receptor, regulates the response to cerebral ischemia by modulating PARP expression and, in turn, apoptosis inhibition by VEGF. In endothelial cell culture, NP-1 colocalized with VEGF receptor-2 (VEGFR-2) and acted as its coreceptor. This significantly enhanced VEGF-induced PARP mRNA and protein expression demonstrated by receptor-specific inhibitors and VEGF-A isoforms. NP-1 augmented the inhibitory effect of VEGF/VEGFR-2 interaction on apoptosis induced by adhesion inhibition through the αV-integrin inhibitor cRGDfV. NP-1/VEGFR-2 signal transduction involved JNK and Akt. In rat models of permanent and temporary middle cerebral artery occlusion, the ischemic cerebral hemispheres displayed endothelial and neuronal apoptosis next to increased endothelial NP-1 and VEGFR-2 expression compared to non-ischemic cerebral hemispheres, sham-operated or untreated controls. Increased vascular superoxide dismutase-1 and catalase expression as well as decreased glycogen reserves indicated oxidative stress in the ischemic brain. Of note, protein levels of intact PARP remained stable despite pro-apoptotic conditions through increased PARP mRNA production during cerebral ischemia. In conclusion, NP-1 is upregulated in conditions of imminent cerebrovascular apoptosis to reinforce apoptosis inhibition and modulate VEGF-dependent PARP expression and activation. We propose that NP-1 is a key modulator of VEGF maintaining cerebrovascular integrity during ischemia. Modulating the function of NP-1 to target PARP could help to prevent cellular damage in cerebrovascular disease., (© 2013.)

Cyclosporine A and FK506 produce immunosuppression by blocking calcineurin phosphatase activity and consequently activation of cytosolic Nuclear Factor of Activated T-cell (NFATc) transcription factor. Due to the chronic toxicity associated with their administration, the development of more specific immunosuppressants is currently an important unmet medical need. In this context, an immunosuppressant peptide derived from the CIC motif of the human Regulators of Calcineurin (RCAN) proteins has been shown to inhibit NFATc signaling without affecting general phosphatase activity of calcineurin. Here we show that protein kinase CK2 phosphorylates a conserved serine residue within the CIC motif of vertebrate RCANs, which increases its affinity for calcineurin and consequently its inhibition of NFATc-dependent gene expression in activated T-cells. Molecular modeling studies have led us to identify a positively charged interaction site on the surface of calcineurin where the phosphorylated serine residue of the CIC motif would normally locate. Finally, we have also identified RCAN3 as a new phosphoprotein with multiple phosphorylation sites. Therefore, our findings reveal for the first time a novel molecular mechanism underlying the regulation of calcineurin-NFATc signaling by means of phosphorylation of the CIC motif of RCAN proteins. The knowledge of how RCAN proteins modulate the calcineurin-NFATc pathway paves the way for the development of potent novel selective immunosuppressant drugs., (Copyright © 2013. Published by Elsevier B.V.)

The family of EF-hand calcium binding proteins is composed of more than 250 members. In search for other neuronal markers, we studied the expression pattern of Necab-1, -2 and -3 in the Ammons horn of adult mice at the gene- and protein levels using in-situ hybridization and immunohistochemistry. The genes for the three Necab's were expressed in specific, non-overlapping areas of the hippocampus. A minority of the Necab-positive interneurons were GABA-ergic, and they virtually never coexpressed one of the classical calcium binding proteins (calretinin, calbindin D-28k and parvalbumin). Necab's are promising new neuronal markers in the brain., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Large cholinergic synaptic boutons called "C-terminals" contact motoneurons and regulate their excitability. C-terminals in the spinal somatic motor nuclei originate from cholinergic interneurons in laminae VII and X that express a transcription factor Pitx2. Cranial motor nuclei contain another type of motoneuron: branchiomotor neurons. Although branchiomotor neurons receive abundant C-terminal projections, the neural source of these C-terminals remains unknown. In the present study, we first examined whether cholinergic neurons express Pitx2 in the reticular formation of the adult mouse brainstem, as in the spinal cord. Although Pitx2-positive cholinergic neurons were observed in the magnocellular reticular formation and region around the central canal in the caudal medulla, none was present more rostrally in the brainstem tegmentum. We next explored the origin of C-terminals in the branchiomotor nuclei by using biotinylated dextran amine (BDA). BDA injections into the magnocellular reticular formation of the medulla and pons resulted in the labeling of numerous C-terminals in the branchiomotor nuclei: the ambiguous, facial, and trigeminal motor nuclei. Our results revealed that the origins of C-terminals in the branchiomotor nuclei are cholinergic neurons in the magnocellular reticular formation not only in the caudal medulla, but also at more rostral levels of the brainstem, which lacks Pitx2-positive neurons., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)