A comparative analysis of pulp-derived nanocelluloses for 3D bioprinting facial cartilages.

Nanocelluloses have attracted significant interest in the field of bioprinting, with previous research outlining the value of nanocellulose fibrils and bacterial nanocelluloses for 3D bioprinting tissues such as cartilage. We have recently characterised three distinct structural formulations of pulp-derived nanocelluloses: fibrillar (NFC), crystalline (NCC) and blend (NCB), exhibiting variation in pore geometry and mechanical properties. In light of the characterisation of these three distinct entities, this study investigated whether these structural differences translated to differences in printability, chondrogenicity or biocompatibility for 3D bioprinting anatomical structures with human nasoseptal chondrocytes. Composite nanocellulose-alginate bioinks (75:25 v/v) of NFC, NCC and NCB were produced and tested for print resolution and fidelity. NFC offered superior print resolution whereas NCB demonstrated the best post-printing shape fidelity. Biologically, chondrogenicity was assessed using real time quantitative PCR, dimethylmethylene blue assays and histology. All biomaterials showed an increase in chondrogenic gene expression and extracellular matrix production over 21 days, but this was superior in the NCC bioink. Biocompatibility assessments revealed an increase in cell number and metabolism over 21 days in the NCC and NCB formulations. Nanocellulose augments printability and chondrogenicity of bioinks, of which the NCC and NCB formulations offer the best biological promise for bioprinting cartilage.

Near-infrared Spectroscopy in Transport With a Patient in Multi-factorial Shock.

A 14-year-old male with a history of repaired truncus arteriosus presented to an outside hospital emergency room in respiratory distress. The triage report to the transport referral center included the following vital signs: temperature of 36.6°C, respiratory rate (RR) of 26 breaths/min, heart rate (HR) of 144 beats/min, and blood pressure (BP) of 113/52 mm Hg with peripheral capillary oxygen saturation (SpO2) of 95% on 4 L via an OxyMask (SouthMedic, Barrie, Ontario, Canada). Additional information indicated severe right ventricle to pulmonary artery conduit stenosis; anuria for 2 days; and cool, mottled extremities. The transport team was dispatched via helicopter. The vital signs upon arrival were as follows: temperature of 36.5°C, HR of 153 beats/min, RR of 48 breaths/min, BP of 81/52, mean arterial pressure of 62, and SpO2 of 96% on 8 L via an OxyMask. Physical assessment revealed the patient was alert and oriented, tachypneic, tachycardic, and displaying poor perfusion. An epinephrine drip was initiated while the patient was being prepared for transport. Near-infrared spectroscopy (NIRS) was initiated with cerebral NIRS of 71% and renal NIRS of 39%. The epinephrine drip was escalated, and norepinephrine was initiated and titrated up for continued poor perfusion and low renal NIRS. Vitals at the transfer of care at the receiving facility were HR of 142 beats/min, BP of 91/51 mm Hg, RR of 56 breaths/min, SpO2 of 99%, and cerebral NIRS of 75% and renal NIRS of 53%. The patient required mechanical circulatory support shortly after admission. NIRS monitoring was used to help measure perfusion and reassess interventions made during transport.

Characterization of pulp derived nanocellulose hydrogels using AVAP® technology.

Bioinspiration from hierarchical structures found in natural environments has heralded a new age of advanced functional materials. Nanocellulose has received significant attention due to the demand for high-performance materials with tailored mechanical, physical and biological properties. In this study, nanocellulose fibrils, nanocrystals and a novel mixture of fibrils and nanocrystals (blend) were prepared from softwood biomass using the AVAP® biorefinery technology. These materials were characterized using transmission and scanning electron microscopy, and atomic force microscopy. This analysis revealed a nano- and microarchitecture with extensive porosity. Notable differences included the nanocrystals exhibiting a compact packing of nanorods with reduced porosity. The NC blend exhibited porous fibrillar networks with interconnecting compact nanorods. Fourier transform infrared spectroscopy and X-ray diffraction confirmed a pure cellulose I structure. Thermal studies highlighted the excellent stability of all three NC materials with the nanocrystals having the highest decomposition temperature. Surface charge analysis revealed stable colloid suspensions. Rheological studies highlighted a dominance of elasticity in all variants, with the NC blend being more rigid than the NC fibrils and nanocrystals, indicating a double network hydrogel structure. Given these properties, it is thought that these materials show great potential in (bio)nanomaterial applications where careful control of microarchitecture, surface topography and porosity are required.

The molecular pharmacology of AMD11070: an orally bioavailable CXCR4 HIV entry inhibitor.

In order to enter and infect human cells HIV must bind to CD4 in addition to either the CXCR4 or the CCR5 chemokine receptor. AMD11070 was the first orally available small molecule antagonist of CXCR4 to enter the clinic. Herein we report the molecular pharmacology of AMD11070 which is a potent inhibitor of X4 HIV-1 replication and the gp120/CXCR4 interaction. Using the CCRF-CEM T cell line that endogenously expresses CXCR4 we have demonstrated that AMD11070 is an antagonist of SDF-1α ligand binding (IC50 = 12.5 ± 1.3 nM), inhibits SDF-1 mediated calcium flux (IC50 = 9.0 ± 2.0 nM) and SDF-1α mediated activation of the CXCR4 receptor as measured by a Eu-GTP binding assay (IC50 =39.8 ± 2.5 nM) or a [(35)S]-GTPγS binding assay (IC50 =19.0 ± 4.1 nM), and inhibits SDF-1α stimulated chemotaxis (IC50 =19.0 ± 4.0 nM). AMD11070 does not inhibit calcium flux of cells expressing CXCR3, CCR1, CCR2b, CCR4, CCR5 or CCR7, or ligand binding to CXCR7 and BLT1, demonstrating selectivity for CXCR4. In addition AMD11070 is able to inhibit the SDF-1ß isoform interactions with CXCR4; and N-terminal truncated variants of CXCR4 with equal potency to wild type receptor. Further mechanistic studies indicate that AMD11070 is an allosteric inhibitor of CXCR4.

Design and synthesis of pyridin-2-ylmethylaminopiperidin-1-ylbutyl amide CCR5 antagonists that are potent inhibitors of M-tropic (R5) HIV-1 replication.

A series of CCR5 antagonists were optimized for potent inhibition of R5 HIV-1 replication in peripheral blood mononuclear cells. Compounds that met acceptable ADME criteria, selectivity, human plasma protein binding, potency shift in the presence of α-glycoprotein were evaluated in rat and dog pharmacokinetics.

Comparing the Ethnic Identity and Well-Being of Adopted Korean Americans with Immigrant/U.S.-Born Korean Americans and Korean International Students.

This study compared the ethnic identity and well-being of Korean Americans who were adopted internationally with immigrant/U.S.-born Korean Americans and Korean international students, as well as the relationship between ethnic identity and well-being for each group. One-hundred and seven college students completed measures of ethnic identity and subjective well-being. Immigrant/U.S.-born Korean Americans had higher ethnic identity scores than the other two groups. Immigrant/U.S.-born Korean Americans also had higher positive affect scores than international students. Ethnic identity was positively correlated with positive affect for all three groups (r's = .27 - .34), but was negatively correlated with negative affect for international students (r = -.44). Overall, the results suggest that ethnic identity, although slightly lower than non-adopted peers, is relevant to the well-being of adopted Korean American college students.

Pharmacology of AMD3465: a small molecule antagonist of the chemokine receptor CXCR4.

CXCR4 is widely expressed in multiple cell types, and is involved in neonatal development, hematopoiesis, and lymphocyte trafficking and homing. Disruption of the CXCL12/CXCR4 interaction has been implicated in stem cell mobilization. Additionally CXCR4 is a co-receptor for HIV. Selective small molecule antagonists of CXCR4 therefore have therapeutic potential. AMD3465 is an N-pyridinylmethylene monocyclam CXCR4 antagonist which can block infection of T-tropic, CXCR4-using HIV. Using the CCRF-CEM T-cell line which expresses CXCR4 we have demonstrated that AMD3465 is an antagonist of SDF-1 ligand binding (K(i) of 41.7+/-1.2nM), and inhibits SDF-1 mediated signaling as shown by inhibition of GTP binding, calcium flux, and inhibition of chemotaxis. AMD3465 is selective for CXCR4 and does not inhibit chemokine-stimulated calcium flux in cells expressing CXCR3, CCR1, CCR2b, CCR4, CCR5 or CCR7, nor does it inhibit binding of LTB(4) to its receptor, BLT1. The pharmacokinetics of AMD3465 was investigated in mice and dogs. Absorption was rapid following subcutaneous administration. AMD3465 was cleared from dog plasma in a biphasic manner with a terminal half-life of 1.56-4.63h. Comparison of exposure to the intravenous and subcutaneous doses indicated 100% bioavailability following subcutaneous administration. AMD3465 caused leukocytosis when administered subcutaneously in mice and dogs, with peak mobilization occurring between 0.5 and 1.5h following subcutaneous dosing in mice and with maximum peak plasma concentration of compound preceding peak mobilization in dogs, indicating that AMD3465 has the potential to mobilize hematopoietic stem cells. These data demonstrate the therapeutic potential for the CXCR4 antagonist AMD3465.

Characterization of the molecular pharmacology of AMD3100: a specific antagonist of the G-protein coupled chemokine receptor, CXCR4.

The chemokine receptor CXCR4 is widely expressed on different cell types, is involved in leukocyte chemotaxis, and is a co-receptor for HIV. AMD3100 has been shown to be a CXCR4 receptor antagonist, and to block HIV infection of T-tropic, X4-using, virus in vitro and in vivo. AMD3100 is an effective mobilizer of hematopoietic stem cells and is being investigated in clinical trials in multiple myeloma and non-Hodgkins lymphoma patients. Using the CCRF-CEM T-cell line that constitutively expresses CXCR4 we confirmed that AMD3100 was an antagonist of SDF-1/CXCL12 ligand binding (IC50=651+/-37 nM). We have also shown that AMD3100 inhibits SDF-1 mediated GTP-binding (IC50=27+/-2.2 nM), SDF-1 mediated calcium flux (IC50=572+/-190 nM), and SDF-1 stimulated chemotaxis (IC50=51+/-17 nM). AMD3100 did not inhibit calcium flux against cells expressing CXCR3, CCR1, CCR2b, CCR4, CCR5 or CCR7 when stimulated with their cognate ligands, nor did it inhibit receptor binding of LTB4. AMD3100 did not, on its own, induce a calcium flux in the CCRF-CEM cells, which express multiple GPCRs including CXCR4, CCR4 and CCR7. Furthermore, AMD3100 neither stimulated GTP-binding, an assay for GPCR activation, in CEM cell membranes; nor chemotaxis of CCRF-CEM cells. These data therefore demonstrate that AMD3100 is a specific antagonist of CXCR4, is not cross-reactive with other chemokine receptors, and is not an agonist of CXCR4.

Imaging of the intracranial venous system with a contrast-enhanced volumetric interpolated examination.

A contrast-enhanced interpolated, three-dimensional (3D) gradient-echo MR sequence with asymmetric k-space sampling, which we refer to as volumetric interpolated brain examination (VIBE), was evaluated for its depiction of the normal intracranial venous system and compared with two-dimensional (2D) time-of-flight (TOF) MR venography (MRV). Fifteen subjects underwent contrast-enhanced VIBE imaging (TR/TE 8 ms/4.4 ms, flip angle 18 degrees, acquisition time, 2 min 20 s, voxel size approximately 1.5 mm(3)) and standard 2D TOF MRV (TR/TE 27 ms/9 ms, flip angle 35 degrees ). The presence of 19 venous structures per subject was assessed on maximum intensity projections (MIP) of the whole data set (whole-brain MIP) and on MIP images reconstructed spontaneously from source images (interactive MIP/source images). Results from a consensus reading where all imaging techniques and display modalities were available were taken as the standard of reference for the presence of venous structures. In addition, 10 subjects underwent both unenhanced and enhanced VIBE imaging. The value of subtracted data sets (unenhanced VIBE subtracted from enhanced VIBE) was then evaluated. Overall, VIBE provided a superior visualization of the cerebral veins than 2D TOF MRV (VIBE, sensitivity (reader 1/reader 2): 98%/99%, negative predictive value 64%/71%; TOF sensitivity: 85%/84%, negative predictive value 15%/15%; Wilcoxon signed-rank test VIBE vs TOF, p<0.001 for both readers). The VIBE interactive MIP/source images were superior to whole-brain MIP reconstructions. Image subtraction was not necessary for delineation of venous structures but improved small vein conspicuity. Contrast-enhanced VIBE acquisitions are faster and enable a visualization of the normal intracranial venous system superior to that of 2D TOF MRV.