310. A Single Skin Contact with Toluene Diiscoyante (Tdi) Causes a One-Year Persistence of Airway Sensitization, Demonstrable in Vivo and in Vitro

Background: [68Ga]Ga-RM2 is a potent Gastrin-Releasing Peptide-receptor (GRP-R) antagonist for imaging prostate cancer and breast cancer, currently under clinical evaluation in several specialized centers around the world. Targeted radionuclide therapy of GRP-R-expressing tumors is also being investigated. We here report the characteristics of a kit-based formulation of RM2 that should ease the development of GRP-R imaging and make it available to more institutions and patients. Methods: Stability of the investigated kits over one year was determined using LC/MS/MS and UV-HPLC. Direct 68Ga-radiolabeling was optimized with respect to buffer (pH), temperature, reaction time and shaking time. Conventionally prepared [68Ga]Ga-RM2 using an automated synthesizer was used as a comparator. Finally, the [68Ga]Ga-RM2 product was assessed with regards to hydrophilicity, affinity, internalization, membrane bound fraction, calcium mobilization assay and efflux, which is a valuable addition to the in vivo literature. Results: The kit-based formulation, kept between 2 °C and 8 °C, was stable for over one year. Using acetate buffer pH 3.0 in 2.5–5.1 mL total volume, heating at 100 °C during 10 min and cooling down for 5 min, the [68Ga]Ga-RM2 produced by kit complies with the requirements of the European Pharmacopoeia. Compared with the module production route, the [68Ga]Ga-RM2 produced by kit was faster, displayed higher yields, higher volumetric activity and was devoid of ethanol. In in vitro evaluations, the [68Ga]Ga-RM2 displayed sub-nanomolar affinity (Kd = 0.25 ± 0.19 nM), receptor specific and time dependent membrane-bound fraction of 42.0 ± 5.1% at 60 min and GRP-R mediated internalization of 24.4 ± 4.3% at 30 min. The [natGa]Ga-RM2 was ineffective in stimulating intracellular calcium mobilization. Finally, the efflux of the internalized activity was 64.3 ± 6.5% at 5 min. Conclusion: The kit-based formulation of RM2 is suitable to disseminate GRP-R imaging and therapy to distant hospitals without complex radiochemistry equipment.

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Characterization of Iron Core–Gold Shell Nanoparticles for Anti-Cancer Treatments: Chemical and Structural Transformations During Storage and Use

Materials ◽

10.3390/ma11122572 ◽

2018 ◽

Vol 11 (12) ◽

pp. 2572 ◽

Cited By ~ 6

Author(s):

Ya-Na Wu ◽

Dar-Bin Shieh ◽

Li-Xing Yang ◽

Hwo-Shuenn Sheu ◽

Rongkun Thordarson ◽

...

Keyword(s):

Au Nanoparticles ◽

Iron Core ◽

Cancer Treatments ◽

Shell Nanoparticles ◽

X Ray ◽

Anti Cancer ◽

One Year ◽

Means Of Transmission

Finding a cancer-selective drug that avoids damaging healthy cells and organs is a holy grail in medical research. In our previous studies, gold-coated iron (Fe@Au) nanoparticles showed cancer selective anti-cancer properties in vitro and in vivo but were found to gradually lose that activity with storage or "ageing.” To determine the reasons for this diminished anti-cancer activity, we examined Fe@Au nanoparticles at different preparation and storage stages by means of transmission electron microscopy combined with and energy-dispersive X-ray spectroscopy, along with X-ray diffraction analysis and cell viability tests. We found that dried and reconstituted Fe@Au nanoparticles, or Fe@Au nanoparticles within cells, decompose into irregular fragments of γ-F2O3 and agglomerated gold clumps. These changes cause the loss of the particles’ anti-cancer effects. However, we identified that the anti-cancer properties of Fe@Au nanoparticles can be well preserved under argon or, better still, liquid nitrogen storage for six months and at least one year, respectively.

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Development and test for long-term stability of a synthetic standard for a quantitative Cryptosporidium parvum LightCycler™ PCR assay

Journal of Water and Health ◽

10.2166/wh.2005.0002 ◽

2005 ◽

Vol 3 (1) ◽

pp. 15-25 ◽

Cited By ~ 1

Author(s):

Renata Filkorn-Kaiser ◽

Konrad Botzenhart ◽

Albrecht Wiedenmann

Keyword(s):

Cryptosporidium Parvum ◽

Surrogate Marker ◽

Target Sequence ◽

Positive Trend ◽

Long Term Stability ◽

Synthetic Standard ◽

One Year

A recently described quantitative rapid cycle real time PCR (LightCycler™) assay detects Cryptosporidium parvum after in vitro excystation, which is a surrogate marker for the viability of the organisms. In the original assay the quantification standard is a dilution series of C. parvum oocysts with a microscopically determined excystation rate. The need to keep suspensions of viable oocysts in stock and to continuously monitor their excystation rate, however, renders the assay impracticable for routine application. A synthetic standard was developed to replace the in vivo standard and was calibrated using oocysts with known excystation rates. The standard consists of a 486 bp DNA segment ranging from 229 bp upstream to 79 bp downstream of the actual PCR target site. Aliquots of the standard were frozen and stored at −20 °C and at −70 °C or lyophilised and stored at room temperature in the dark. For a period of one year samples preserved with each of the three methods were restored every four or five weeks. They were amplified in the LightCycler™ and the crossing points (CP) were monitored. No significant trend in the raw CP values could be observed for any of the three storage methods. However, when the methods were compared to each other by calculating the CP ratios (−20 °C/−70 °C; −20 °C/lyophilised; −70 °C/lyophilised) at the 10 monitoring dates, the CP ratios −20 °C/−70 °C and −20 °C/lyophilised showed a highly significant positive trend (p<0.0001) while the CP ratio −70 °C/lyophilised did not differ from the null hypothesis (p=0.53). It can be concluded that the latter two preservation methods are both appropriate, while storage at −20 °C is less advisable. Calculations based on the molecular weight of the standard and on the assumption of an average yield of three sporozoites per oocyst led to the conclusion that the target sequence is probably located on a double copy gene

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A Review of Multimatrix System (MMX) Mesalazine in the Management of Ulcerative Colitis

Clinical Medicine Therapeutics ◽

10.4137/cmt.s38 ◽

2009 ◽

Vol 1 ◽

pp. CMT.S38

Author(s):

A Barney Hawthorne

Keyword(s):

High Strength ◽

Mucosal Healing ◽

In Vivo Studies ◽

Systemic Absorption ◽

Pharmacokinetic Data ◽

Once Daily ◽

Maintenance Of Remission ◽

One Year

MMX™ mesalazine is a novel delayed release mesalazine formulation with a high-strength 1.2 g tablet for the treatment of active mild to moderate colitis and maintenance of remission. In vitro and in vivo studies show initiation of drug release in the terminal ileum and caecum with gradual release of 5-ASA as the tablet passes through the colon. Pharmacokinetic data are comparable to other 5-ASA formulations with low systemic absorption and high levels in feces and in mucosal biopsies in the left colon. Clinical trials have shown high rates of clinical and endoscopic remission in active mild to moderate colitis over 8 weeks with a 2.4 g once daily dose. There do not appear to be higher remission rates with the 4.8 g dose. Prolongation of treatment with a further 8 weeks of 2.4 g twice daily can induce remission in those failing the initial 8 weeks of therapy. A maintenance study enrolling patients who achieved remission in the acute studies showed high rates of remission maintained at one year with 1.2 g twice daily (68.5%) and 2.4 g once daily (64.4%) using the strict definition of remission (including mucosal healing) that was used in the active treatment trials. The drug is safe and effective in colitis. The high tablet strength, and once-daily dosage make this formulation a welcome addition to therapy options for patients with colitis.

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Vitis Methods to Understand and Develop Strategies for Diagnosis and Sustainable Control of Grapevine Trunk Diseases

Phytopathology ◽

10.1094/phyto-09-18-0349-rvw ◽

2019 ◽

Vol 109 (6) ◽

pp. 916-931 ◽

Cited By ~ 6

Author(s):

P. Reis ◽

R. Pierron ◽

P. Larignon ◽

P. Lecomte ◽

E. Abou-Mansour ◽

...

Keyword(s):

Single Agent ◽

Gray Mold ◽

Economic Losses ◽

Advantages And Disadvantages ◽

Grapevine Trunk Diseases ◽

Trunk Diseases ◽

One Year ◽

Integrative Levels

Vitis vinifera is affected by many diseases every year, depending on causal agents, susceptibility of cultivars, and climate region. Some are caused by a single agent, such as gray mold caused by Botrytis cinerea or powdery mildew caused by Erysiphe necator. Others result from the actions of a complex of pathogens such as grapevine trunk diseases (GTDs). GTDs are presently among the most devastating diseases in viticulture worldwide because both the economic losses and the long-term sustainability of vineyards are strongly affected. The complexity of GTDs results from the diversity of associated fungi, the undetermined period of latency within the vine (asymptomatic status), the erratic foliar symptom expression from one year to the next, and, probably correlated with all of these points, the lack of efficient strategies to control them. Distinct methods can be beneficial to improve our knowledge of GTDs. In vitro bioassays with cell suspensions, calli, foliar discs, full leaves, or plantlets, and in vivo natural bioassays with cuttings, grafted plants in the greenhouse, or artificially infected ones in the vineyard, can be applied by using progressive integrative levels of in vitro and in vivo, depending on the information searched. In this review, the methods available to understand GTDs are described in terms of experimental procedures, main obtained results, and deliverable prospects. The advantages and disadvantages of each model are also discussed.

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39. CHARACTERIZING NOVEL INHIBITORS OF BRAIN METASTASIS-INITIATING CELLS

Neuro-Oncology Advances ◽

10.1093/noajnl/vdaa073.027 ◽

2020 ◽

Vol 2 (Supplement_2) ◽

pp. ii7-ii7

Author(s):

Agata Kieliszek ◽

Chitra Venugopal ◽

Blessing Bassey-Archibong ◽

Nikoo Aghaei ◽

Fred Lam ◽

...

Keyword(s):

At Risk ◽

Brain Metastasis ◽

Primary Tumor ◽

Unmet Need ◽

Standard Of Care ◽

Current Standard ◽

One Year ◽

Pdx Models

Abstract BACKGROUND The incidence of brain metastases (BM) is tenfold higher than primary brain tumors. BM commonly originate from primary lung, breast, and melanoma tumors with a 90% mortality rate within one year of diagnosis. Current standard of care for BM includes surgical resection with concurrent chemoradiation, but does not extend median survival past 16 months, posing a large unmet need to identify novel therapies against BM. METHODS From a large in-house biobank of patient-derived BM cell lines, the Singh Lab has generated murine orthotopic patient-derived xenograft (PDX) models of lung, breast, and melanoma BM that recapitulate the stages of BM progression as seen in human patients. Using these three PDX models, we identified a population of “pre-metastatic” brain metastasis-initiating cells (BMICs) that are newly arrived in the brain but have yet to form detectable tumors. Pre-metastatic BMICs are not detectable in human patients but are important therapeutic targets with the potential to prevent BM in at-risk patients. RESULTS RNA sequencing of pre-metastatic BMICs from all three PDX primary tumor models with subsequent Connectivity Map analysis identified novel compounds that have the potential of killing all three types of BMICs. In particular, we identified two compounds that have selective killing of BMICs in vitro from all three primary tumor cohorts while sparing non-cancerous cells. We further characterized their ability to inhibit the self-renewal and proliferative properties of BMICs. Ongoing in vivo work will investigate the compounds’ preclinical utilities in preventing BM. CONCLUSION Identification of novel small molecules that target BMICs could prevent the formation of BM completely and dramatically improve the prognosis of at-risk cancer patients.

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Bacteriostasis of Escherichia coli by milk. V. The bacteriostatic properties of milk of West African mothers in The Gambia: in-vitro studies

Journal of Hygiene ◽

10.1017/s0022172400063427 ◽

1980 ◽

Vol 85 (3) ◽

pp. 347-358 ◽

Cited By ~ 7

Author(s):

Jean M. Dolby ◽

Pauline Honour ◽

M. G. M. Rowland

Keyword(s):

High Activity ◽

Similar Degree ◽

Bacteriostatic Activity ◽

E Coli ◽

Highly Active ◽

High Incidence ◽

Iron Binding Protein ◽

One Year

SUMMARYBacteriostatic activity was measured in 244 specimens of milk collected during 1977 throughout lactation of up to one year from 78 mothers; the activity varied from very good to fair and only seven were inactive. There was a wider range of activity than was found previously in milk from English mothers. Activity usually fell slowly during lactation but some of the Gambian mothers produced milk of very high activity, like that of colostrum into the second week of lactation, and two mothers did so at six and nine months; other mothers produced good-activity milk throughout lactation. The bacteriostatic activity varied little with the season but slight decreases from that expectcd were found after the high incidence of infant diarrhoea towards the end of the rainy season.The bacteriostatic activity of most of the milk tested could be prevented by iron salts but that of colostrum and some of the milks with high activity could not. Only these highly active colostra and milks were inhibitory in vitro when the inoculum was increased from 104 to 106 organisms per ml. These and less active milks were able to inhibit the smaller, standard inoculum for longer than 3 h with the addition of bicarbonate and extra iron-binding protein at the concentrations likely to be present in vivo. Both commensal and pathogenic E. coli were inhibited to a similar degree by these milks and there was no evidence of serotype specificity.

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In Vitro Glioblastoma Models: A Journey into the Third Dimension

Cancers ◽

10.3390/cancers13102449 ◽

2021 ◽

Vol 13 (10) ◽

pp. 2449

Author(s):

Mayra Paolillo ◽

Sergio Comincini ◽

Sergio Schinelli

Keyword(s):

Tumor Cells ◽

Single Cell Analysis ◽

3D Models ◽

Primary Brain Tumor ◽

Molecular Features ◽

Third Dimension ◽

One Year ◽

The Impact

Glioblastoma multiforme (GBM) is the most lethal primary brain tumor in adults, with an average survival time of about one year from initial diagnosis. In the attempt to overcome the complexity and drawbacks associated with in vivo GBM models, together with the need of developing systems dedicated to screen new potential drugs, considerable efforts have been devoted to the implementation of reliable and affordable in vitro GBM models. Recent findings on GBM molecular features, revealing a high heterogeneity between GBM cells and also between other non-tumor cells belonging to the tumoral niche, have stressed the limitations of the classical 2D cell culture systems. Recently, several novel and innovative 3D cell cultures models for GBM have been proposed and implemented. In this review, we first describe the different populations and their functional role of GBM and niche non-tumor cells that could be used in 3D models. An overview of the current available 3D in vitro systems for modeling GBM, together with their major weaknesses and strengths, is presented. Lastly, we discuss the impact of groundbreaking technologies, such as bioprinting and multi-omics single cell analysis, on the future implementation of 3D in vitro GBM models.

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Transcriptional modulation of Interleukin-11 by the coronaviruses.

10.31219/osf.io/r4syv ◽

2020 ◽

Author(s):

Shahan Mamoor

Keyword(s):

Differential Expression ◽

Differentially Expressed ◽

The Novel ◽

Transcriptional Modulation ◽

One Year ◽

Microarray Datasets ◽

Interleukin 11 ◽

In Cells

The novel SARS-CoV-2 has infected nearly 20,000,000 people worldwide in less than one year (1). We mined published and public microarray datasets (2-7) to identify genes most differentially expressed in cells and tissues infected with a number of coronavirus. We describe differential expression of the cytokine interleukin-11 following infection with MERS-CoV in vitro and SARS-CoV-1 in vivo.

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An Implantable Microfabricated Drug Delivery System

Applied Mechanics and Biomedical Technology ◽

10.1115/imece2003-43186 ◽

2003 ◽

Cited By ~ 2

Author(s):

John M. Maloney

Keyword(s):

Drug Delivery ◽

Drug Delivery System ◽

Delivery System ◽

In Vivo Studies ◽

Multiple Drug ◽

Hermetic Storage ◽

Continuous Release ◽

One Year

We report on the development of a fully implantable drug delivery system capable of delivering hundreds of individual doses. This product is intended for the controlled release of potent therapeutic compounds that might otherwise require frequent injections. Our system has the following capabilities: • Stable, hermetic storage of therapeutic drugs in solid, liquid, or gel form; • Individual storage of discrete doses for multiple-drug regimens; • Wireless communication with an external controller for device monitoring and therapy modification; • Choice of preprogrammed release or release on command; • Controlled pulsatile or continuous release. MicroCHIPS’ drug release technology has been successfully demonstrated in vitro and in vivo. We are proceeding with long-term in vivo studies of a fully implantable device containing one hundred individual doses. A future device intended for human clinical trials will contain four hundred doses, enough for a daily release of drug for more than one year.

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