The Promise and Peril of Chemical Probe Negative Controls

2021 ◽  
Author(s):  
Jinyoung Lee ◽  
Matthieu Schapira
Keyword(s):  
Chemical Probe ◽  
Negative Controls

Cellular Localization of Enzymatically Active Thrombin in Intact Human Tissues by Hirudin Binding

1995 ◽  
Vol 73 (05) ◽  
pp. 793-797 ◽  
Author(s):  
Leo R Zacharski ◽  
Vincent A Memoli ◽  
William D Morain ◽  
Jean-Marc Schlaeppi ◽  
Sandra M Rousseau
Keyword(s):  
Tumor Cell ◽  
Cell Carcinoma ◽  
Cellular Localization ◽  
Thrombin Generation ◽  
Normal Lung ◽  
Cancer Tissue ◽  
Immunohistochemical Techniques ◽  
Tumor Types ◽  
Negative Controls

SummaryCellular sites of coagulation activation within complex, intact tissues have been studied by immunohistochemical techniques. Hirudin, a specific and high affinity inihibitor of the active site of thrombin, together with antibody to hirudin were applied to sections of AMeX-fixed specimens of normal lung, kidney, placenta, freshly incised skin and unperturbed skin obtained at fresh autopsy; to rheumatoid synovial tissue; and to malignant tissue from a variety of tumor types. Staining for thrombin was observed selectively on pulmonary alveolar, rheumatoid synovial, and placental macrophages that express an intact extrinsic coagulation pathway. Staining was also observed restricted to the endothelium of capillaries in freshly incised skin but not in either unperturbed skin or in aged incisions. Staining of tumor cell bodies was observed in small cell carcinoma of the lung, renal cell carcinoma, and malignant melanoma tissues that we found previously to show tumor cell-associated procoagulant activity. This staining occurred commonly on cells within the tumor mass that were distant from stromal fibrinogen/fibrin. By contrast, tumor-associated macrophage but not tumor cell staining was seen in adenocarcinoma and squamous cell carcinoma of the lung, and little or no staining was seen in colon cancer tissue. Negative controls in which either the hirudin probe or its antibody were omitted failed to show staining. These results are in accord with previous findings and suggest that such techniques may be useful for studying the cellular sites of thrombin generation in intact tissues. We postulate that administration of potent and specific thrombin antagonists, such as hirudin, to patients with relevant tumor types might be followed by homing of hirudin to tumor cells in vivo so that effects of local thrombin generation on malignant progression can be determined.

SGC-CK2-1: The First Selective Chemical Probe for the Pleiotropic Kinase CK2

2020 ◽  
Author(s):  
Carrow Wells ◽  
David Drewry ◽  
Julie E. Pickett ◽  
Alison D. Axtman
Keyword(s):  
Cell Lines ◽  
Small Molecule ◽  
Chemical Probe ◽  
High Quality ◽  
Extensive Evaluation ◽  
Key Driver ◽  
Selective Chemical ◽  
Kinase Ck2 ◽  
Ck2 Inhibitors

Building upon a wealth of published knowledge surrounding the pyrazolopyrimidine scaffold, we designed a small library around the most selective small molecule CK2 inhibitors reported. Through extensive evaluation of this library we identified inhibitor 24 (SGC-CK2-1) as a potent, selective, and cell-active CK2 chemical probe. Remarkably, despite years of research pointing to CK2 as a key driver in cancer, our probe did not elicit an antiproliferative phenotype in cell lines tested. While many publications have attempted tocharacterize CK2 function, CK2 biology is complex and a high-quality chemical tool like SGC-CK2-1 will aid in connecting CK2 functions to phenotypes.

Validity of a Serological Diagnostic Kit for SARS-CoV-2 Available in Iran

2020 ◽  
Vol 23 (9) ◽  
pp. 629-632
Author(s):  
Hamid Reza Shamsollahi ◽  
Mostafa Amini ◽  
Shaban Alizadeh ◽  
Saharnaz Nedjat ◽  
Ali Akbari-Sari ◽  
...  
Keyword(s):  
Diagnostic Techniques ◽  
Effective Control ◽  
Molecular Technique ◽  
Size Estimation ◽  
Medical Sciences ◽  
Serological Tests ◽  
Standard Case ◽  
Epidemic Size ◽  
Low Sensitivity ◽  
Negative Controls

Background: The Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemic broke out in December 2019 and is now characterized as a pandemic. Effective control of this infectious disease requires access to diagnostic techniques, for both case finding and epidemic size estimation. The molecular technique is routinely used worldwide. Although it is the "standard" case detection and management method, it has its own shortcomings. Thus, some easy-to-use rapid serological tests have been developed. Methods: One hundred and fourteen positive RT-PCR-diagnosed patients were tested by VivaDiag Kit, a brand of rapid serological kits available in hospitals affiliated to Tehran University of Medical Sciences (TUMS), Tehran, Iran. Frozen serum specimens taken from healthy people in summer and fall 2019 were also tested as negative controls. Results: Test sensitivity was 47.9% (95% confidence interval [CI]: 38.8-56.9) for IgM and 47.0% (95% CI: 38.0–56.0) for IgG. There was no difference between IgG and IgM seropositivity except in one case. Specificity was calculated as 99.0% (95% CI: 96.4–99.9) for IgM and of 100.0% (95% CI: 0.98.2–100.0) for IgG. Sensitivity was higher in men and older participants. Conclusion: This test can be used for epidemiological investigations, especially for the estimation of the level of infection in the community, after it is properly corrected for sensitivity and specificity. The low sensitivity could be attributed to the technical limitations of the kit or low levels of antibodies after infection. The different sensitivity in age and sex groups supports the hypothesis that different people show different immune responses to this virus.

Bevel Structure Based XPS Analysis as a Non‐Destructive Chemical Probe for Complex Interfacial Structures of Organic Semiconductors

Small Methods ◽  
2021 ◽  
pp. 2001264
Author(s):  
Dong‐Jin Yun ◽  
Seunghyup Lee ◽  
Seong Heon Kim ◽  
Changhoon Jung ◽  
Yong Su Kim ◽  
...  
Keyword(s):  
Organic Semiconductors ◽  
Xps Analysis ◽  
Chemical Probe ◽  
Interfacial Structures ◽  
Non Destructive

scholarly journals The course of subjective and objective chemosensory dysfunction in hospitalized patients with COVID-19: a 6-month follow-up

2021 ◽  
Author(s):  
Mattis Bertlich ◽  
Clemens Stihl ◽  
Enzo Lüsebrink ◽  
Johannes C. Hellmuth ◽  
Clemens Scherer ◽  
...  
Keyword(s):  
Long Term Effects ◽  
Initial Examination ◽  
Initial Cohort ◽  
Negative Controls ◽  
Chemosensory Function ◽  
Analogue Scales ◽  
Smell Identification ◽  
Snot 22

Abstract Purpose It has been established that the infection with SARS-CoV-2 may cause an impairment of chemosensory function. However, there is little data on the long-term effects of SARS-CoV-2 infection on chemosensory function. Methods Twenty three SARS-CoV-2-positive patients diagnosed in spring 2020 with subjective hyposmia (out of 57 positive patients, 40.3%) were compared to SARS-CoV-2-positive patients without hyposmia (n = 19) and SARS-CoV-2-negative patients (n = 14). Chemosensory function was assessed by the Brief Smell Identification Test (BSIT), Taste Strips (TS), Visual Analogue Scales (VAS), and the SNOT-22. The initial cohort with hyposmia were also examined at 8 weeks and 6 months after initial examination. Results There were no differences between the SARS-CoV-2-positive cohort without hyposmia and negative controls in terms of BSIT (8.5 ± 2.6 vs. 10.2 ± 1.8), TS (3.4 ± 0.6 vs. 3.9 ± 0.3) or VAS (2.1 ± 1.3 vs. 1.1 ± 0.5); yet the SNOT-22 was significantly elevated (27.7 ± 11.2 vs. 16.4 ± 10.8). The SARS-CoV-2-positive group with hyposmia performed significantly poorer in BSIT (4.0 ± 1.7 vs. 8.5 ± 2.6/10.2 ± 1.8), TS (2.6 ± 1.3 vs. 3.4 ± 0.6/3.9 ± 0.3), and VAS (7.9 ± 2.2 vs. 2.1 ± 1.3/1.1 ± 0.5) compared to both control groups. At week 8 and month 6 control, six and five patients, respectively, still suffered from subjectively and objectively impaired chemosensory function. The other patients had recovered in both respects. Conclusion SARS-CoV-2 patients with subjectively impaired chemosensory function regularly perform poorly in objective measurements. About 70% of patients suffering from olfactory dysfunction in SARS-CoV-2 quickly recover—the rest still suffers from considerable impairment 6 months after infection.

scholarly journals Macrophagic Inflammatory Response Next to Dental Implants with Different Macro- and Micro-Structure: An In Vitro Study

2021 ◽  
Vol 11 (12) ◽  
pp. 5324
Author(s):  
Maria Menini ◽  
Francesca Delucchi ◽  
Domenico Baldi ◽  
Francesco Pera ◽  
Francesco Bagnasco ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Dental Implants ◽  
In Vitro Study ◽  
Titanium Implants ◽  
Implant Surface ◽  
Bone Implant ◽  
Optimal Outcomes ◽  
Negative Controls ◽  
Inflammatory Effect

(1) Background: Intrinsic characteristics of the implant surface and the possible presence of endotoxins may affect the bone–implant interface and cause an inflammatory response. This study aims to evaluate the possible inflammatory response induced in vitro in macrophages in contact with five different commercially available dental implants. (2) Methods: one zirconia implant NobelPearl® (Nobel Biocare) and four titanium implants, Syra® (Sweden & Martina), Prama® (Sweden & Martina), 3iT3® (Biomet 3i) and Shard® (Mech & Human), were evaluated. After 4 h of contact of murine macrophage cells J774a.1 with the implants, the total RNA was extracted, transcribed to cDNA and the gene expression of the macrophages was evaluated by quantitative PCR (qPCR) in relation to the following genes: GAPDH, YWHAZ, IL1β, IL6, TNFα, NOS2, MMP-9, MMP-8 and TIMP3. The results were statistically analyzed and compared with negative controls. (3) Results: No implant triggered a significant inflammatory response in macrophages, although 3iT3 exhibited a slight pro-inflammatory effect compared to other samples. (4) Conclusions: All the samples showed optimal outcomes without any inflammatory stimulus on the examined macrophagic cells.

scholarly journals Nanocarriers of Miconazole or Fluconazole: Effects on Three-Species Candida Biofilms and Cytotoxic Effects In Vitro

2021 ◽  
Vol 7 (7) ◽  
pp. 500
Author(s):  
Anne Caroline Morais Caldeirão ◽  
Heitor Ceolin Araujo ◽  
Laís Salomão Arias ◽  
Wilmer Ramírez Carmona ◽  
Gustavo Porangaba Miranda ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Artificial Saliva ◽  
Cytotoxic Effects ◽  
Promising Alternative ◽  
Colony Forming Units ◽  
Mtt Reduction ◽  
Diphenyl Tetrazolium Bromide ◽  
Negative Controls ◽  
Positive Controls

The contribution of different Candida species in oral fungal infections has stimulated the search for more effective therapies. This study assessed the antibiofilm effects of nanocarriers of miconazole (MCZ) or fluconazole (FLZ) on Candida biofilms, and their cytotoxic effects on murine fibroblasts. Three-species biofilms (Candida albicans/Candida glabrata/Candida tropicalis) were formed on 96-well plates, and they were treated with nanocarriers (iron oxide nanoparticles coated with chitosan—“IONPs-CS”) of MCZ or FLZ at 39/78/156 µg/mL; antifungals alone at 156 µg/mL and artificial saliva were tested as positive and negative controls, respectively. Biofilms were analyzed by colony forming units (CFU), biomass, metabolic activity, and structure/viability. The cytotoxicity (L929 cells) of all treatments was determined via 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) reduction assay. Data were submitted to one- or two-way ANOVA, followed by Tukey’s or Fisher LSD’s tests (p < 0.05). IONPs-CS-MCZ at 78 µg/mL promoted similar antibiofilm and cytotoxic effects compared with MCZ at 156 µg/mL. In turn, IONPs-CS-FLZ at 156 µg/mL was overall the most effective FLZ antibiofilm treatment, surpassing the effects of FLZ alone; this nanocarrier was also less cytotoxic compared with FLZ alone. It can be concluded that both nanocarriers are more effective alternatives to fight Candida biofilms compared with their respective positive controls in vitro, being a promising alternative for the treatment of oral fungal infections.

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