References

Lund K, Olsen CE, Wong JJW, Olsen PA, Solberg NT, Høgset A, Krauss S, Selbo PK (2017) 5-FU resistant EMT-like pancreatic cancer cells are hypersensitive to photochemical internalization of the novel endoglin-targeting immunotoxin CD105-saporin. J Exp Clin Cancer Res 36(1):185.. doi: 10.1186/s13046-017-0662-6

Objective: Investigate resistance mechanisms and photochemical strategies to overcome 5-FU resistance in pancreatic adenocarcinoma.

Summary: Expression of CD105 was investigated using RT-qPCR, western blotting, flow cytometry, and fluorescence microscopy, and co-localization of TPCS2a and Anti-CD105-SAP was assessed using microscopy. MTS assay was used to investigate cytotoxic effects of photochemical internalization of Anti-CD105-SAP. For the first time, we demonstrate the promise of PCI-based targeting of CD105 in site-specific elimination of 5-FU resistant pancreatic cancer cells using Anti-CD105-SAP in vitro. PCI-based targeting of CD105 may represent a potent anti-cancer strategy and should be further evaluated in preclinical models.

Usage: Cells were seeded (3000/well) in 96-well plates and allowed to attach overnight. The cells were incubated with the Anti-CD105-saporin (2.4 nM) or Saporin as a control (6.48 nM; Saporin was added in a molecular ratio of 2.7:1 to the immunotoxin) giving an equal ratio of Saporin to immunotoxin), with or without the photosensitizer TPCS2a (0.35 μg/ml) for 18 h.

Related Products: Anti-CD105-SAP (Cat. #IT-80)

Thom G, Hatcher J, Hearn A, Paterson J, Rodrigo N, Beljean A, Gurrell I, Webster C (2018) Isolation of blood-brain barrier-crossing antibodies from a phage display library by competitive elution and their ability to penetrate the central nervous system. mAbs 10:304-314. doi: 10.1080/19420862.2017.1409320 PMID: 29182455

Summary: Fluorescence microvolume assay technology (FMAT) periprep extract or Nickel-purified scFvs from E. coli were transferred into assay plates containing 10 ml (1:1000 dilution of a 1 mg/ml stock solution) mouse anti-HIS MAb.)

Related Products: 6His Mouse Monoclonal (Cat. #AB-213)

Ullah K, Rosendahl AH, Izzi V, Bergmann U, Pihlajaniemi T, Maki JM, Myllyharju JA-Ohoo (2017) Hypoxia-inducible factor prolyl-4-hydroxylase-1 is a convergent point in the reciprocal negative regulation of NF-κB and p53 signaling pathways. Sci Rep 7:172200. doi: 10.1038/s41598-017-17376-0 PMID: 29222481

Usage: Western blot

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Brick RM, Sun AX, Tuan RSA-Ohoo (2018) Neurotrophically induced mesenchymal progenitor cells derived from induced pluripotent stem cells enhance neuritogenesis via neurotrophin and cytokine production. Stem Cells Transl Med 7:45-58. doi: 10.1002/sctm.17-0108 PMID: 29215199

Objective: To examine the potential utility of an alternative, mesenchymal-like cell (MSCs) source, derived from induced pluripotent stem cells, termed induced mesenchymal progenitor cells (MiMPCs) to produce bioactive factors and support nerve regeneration.

Summary: The findings suggest MiMPCs as a renewable, candidate source of therapeutic cells and a potential alternative to MSCs for peripheral nerve repair, in view of their ability to promote nerve growth by producing many of the same growth factors and cytokines as Schwann cells and signaling through critical neurotrophic pathways.

Usage: Immunofluorescence

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Zhang J, Wang H, Wu S, Liu Q, Wang N (2017) Regulation of reentrainment function is dependent on a certain minimal number of intact functional iprgcs in rd mice. J Ophthalmol 2017:6804853.. doi: 10.1155/2017/6804853

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Khan N, Muralidharan A, Smith MT (2017) Attenuation of the infiltration of angiotensin ii expressing CD3+ T-cells and the modulation of nerve growth factor in lumbar dorsal root ganglia – a possible mechanism underpinning analgesia produced by EMA300, an Angiotensin II type 2 (AT2) receptor antagonist. Front Mol Neurosci 10:389. doi: 10.3389/fnmol.2017.00389 PMID: 29200998

Objective: To investigate the cellular and molecular mechanism of action of selective small molecule angiotensin II type 2 (AT2) receptor antagonists in the alleviation of peripheral neuropathic pain.

Summary: The analgesic effect of EMA300 in CCI-rats involves multimodal actions that appear to be mediated at least in part by a significant reduction in the otherwise increased expression levels of Ang II as well as the number of Ang II-expressing CD3+ T-cells in the ipsilateral lumbar DRGs of CCI-rats.

Usage: Immunocytochemistry; specifically labeled HEK cells expressing the AT2 but not the AT1 receptor or the non-transfected HEK-cells.

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Liu A, Aoki S, Wickens J (2017) A streamlined method for the preparation of gelatin embedded brains and simplified organization of sections for serial reconstructions. Bio-protocol 7(22):e2610.. doi: 10.21769/BioProtoc.2610

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Lovatt M, Yam GH-F, Peh GS, Colman A, Dunn NR, Mehta JS (2018) Directed differentiation of periocular mesenchyme from human embryonic stem cells. Differentiation 99:62-69. doi: 10.1016/j.diff.2017.11.003 PMID: 29239730

Objective: Pluripotent stem cells are attractive sources of cells for regenerative medicine, because large numbers of therapeutically useful cells can be generated. However, a detailed understanding of how to differentiate clinically relevant cell types from stem cells is fundamentally required.

Summary: Identification of cells resembling periocular mesenchyme (POM) cells in the adult cornea, located in a niche between the trabecular meshwork and peripheral endothelium. The generation and expansion of POM is an important step in the generation of a number of cells types that could prove to be clinically useful for a number of diseases of the cornea.

Usage: 1:200 for flow cytometry and immunofluorescence.

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Horner KA, Logue JB, Jenrette TA (2017) Patch compartment lesions reduce habitual sucrose consumption. Neuroscience 2017 Abstracts 689.16 / II23. Society for Neuroscience, Washington, DC.

Summary: The striatum mediates habit formation and reward association. The striatum can be divided into the patch and matrix compartment, which are two neurochemically and anatomically distinct regions that may sub-serve different aspects of behavior. For example, the patch compartment may mediate reward-related behaviors, while the matrix compartment may mediate adaptive motor functions. Furthermore, previous studies have shown that enhanced relative activation of the patch versus matrix compartment is associated with inflexible behaviors, such as stereotypy. Habitual behaviors are also inflexible in nature, but whether enhanced activation of the patch compartment contributes to habitual behavior is not known. The goal of the current study was to examine the role of patch compartment neurons in the development of habit formation. We used dermorphin-saporin to specifically ablate neurons of the patch compartment prior to training animals to self-administer sucrose on a random interval schedule of reinforcement, which has been shown to foster habit formation. Our data showed that destruction of the neurons of the patch compartment prevented the reinstatement of sucrose self-administration after sucrose devaluation, indicating that absence of the patch compartment interrupted the development of habitual behavior. Our data also indicate that c-Fos levels were decreased in the dorsolateral striatum (DLS) and sensorimotor cortex (SMC), but increased in dorsomedial striatum (DMS) and prefrontal cortex (PFC) in patch-lesioned animals that did not develop habitual behavior, indicating that diminished habit formation is associated with decreased activation of regions that participate in habitual behavior, and increased in regions associated with goal-directed behaviors. Together, these data indicate that the patch compartment participates in habit formation by altering the flow of information through basal ganglia circuits.

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Falquetto B, Oliveira LM, Moreira TS, Takakura AC (2017) Role of orexinergic neurons in the chemosensory control of breathing in a Parkinson’s disease model. Neuroscience 2017 Abstracts 779.08 / HH1. Society for Neuroscience, Washington, DC.

Summary: Parkinson´s disease (PD) is a neurodegenerative disorder characterized by progressive loss of dopaminergic neurons in the substantia nigra compacta (SNpc). Non-motor symptoms such as neuropsychiatric, sleep and breathing disorders are also observed in PD. Previous study has already demonstrated that in 6-hydroxydopamine (6-OHDA)-model of PD there is a reduction in the number of phox2b neurons in the retrotrapezoid nucleus (RTN) and a decrease in the respiratory response to hypercapnia. Here, we tested the involvement of orexin cells from lateral hypothalamus/perifornical area (LH/PeF) on breathing in this model of PD. 6-OHDA (24 µg/µl) injections into the striatum reduced the number of catecholaminergic (40 days: 128 ± 10 and 60 days: 116 ± 13 vs. vehicle: 938 ± 15 neurons) and orexin-B-ir neurons (40 days: 310 ± 9 and 60 days: 258 ± 15 vs. vehicle: 412 ± 13 neurons). The injection of anti-Orexin-B saporin into the LH/PeF produces a further reduction in the number of orexinergic neurons in PD animals (79 ± 8 vs. control: 427 ± 14 neurons). The respiratory frequency (fR) at rest and in response to hypercapnia (7% CO2) was assessed 60 days after bilateral 6-OHDA or vehicle injections into the striatum and anti-Orexin-B saporin or IgG saporin into the LH/PeF during sleep and wakefulness in the dark and light phases of the diurnal cycle. Sixty days after 6-OHDA, we observed a reduction of fR at rest during sleep in the light phase only in PD animals (56 ± 2 vs. control: 66 ± 2 bpm). During the dark phase, there is a reduction in fR response to hypercapnia in PD animals with depletion of orexinergic neurons during wakefulness (119 ± 6 vs. control: 152 ± 3 bpm) and sleep (128 ± 7 vs. control: 147 ± 5 bpm). Our data suggest that orexinergic neurons are important to restore chemoreceptor function in a rat model of PD during sleep and wakefulness in rats.

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