human prostate carcinoma cell line du145  (ATCC)

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 1. Nude nu/numale mice groups treatment after implanted DU145 cells

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques:

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 2. Mortality of mice implanted with DU145 cells as groups I, II, III (treated by VK3:VC) and IV (control untreated).

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Control

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 3. a-e: pane with LM photomontages of DU145 ×enografts established on nu/nu mouse diaphragm one µm-thick, toluidine blue stained epoxy sections (a, b and d), SEM (c), and TEM (e) views. Early (a) to expanded size (b, d) and epithelial surface (c) show also the pleiomorphic population, including one TEM example of the active DU145 cells (in e) showing a huge branched nucleolar organizer center (NOR). Stars marked examples of scarce cytophagocytosis; black arrows marked small cells, and V for vascular components surrounded by connective cells.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Staining

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 4. a-e: pane with LM photomontages one µm-thick toluidine blue stained epoxy sections, and SEM of DU145 ×enografts grown on nu/nu mouse diaphragm (d) treated by oral (a and d), intraperitoneal (IP, in b), and by both oral and IP (c) VK3:VC combined. In the lower region, the installation on remnants of the diaphragm (d) and epimysium (open arrows in e) showed the carcinomas with cryptic demolitions as autolyzes (stars), mitotic cell arrest in telophase (te); v: vascular structures. In c: SEM aspect of d marked disrupted epithelial surface.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Staining

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 5. a-b: TEM fields of view of a larger than 200 µm-thick DU145 carcinoma after oral VK3:VC treatment. Most cells care altered throughout the tumor showing nucleus degenerations and cytoplasmic vacuolizations, comforting those noted in Figures 4(a-c), and those in Figures 6–17. Figures 5(c-d): comparative TEM aspect of untreated DU145 cell in prophase (c) and with enlarged chromosome parts (d).

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques:

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 6. TEM and SEM aspects after DU145 tumor oral VK3: VC treatment shows an altered surface epithelium with some remained microvilli as also noted with SEM (insert). Other disrupted intercellular contact junctions, diverse organelle swellings, included remnants of mitochondria (mt), lipid droplets (li), phagosomes (ph), and cell debris (small arrows); a contrasted, unidentified “belt” (c; connective ground substance?) appeared to join adjacent damaged cells. A remnant of cell corpse vacuolated is at d.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques:

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 7. TEM aspect of IP VK3:VC-treated mice bearing a DU145 carcinoma. Inner tissue location with cells alterations. Nucleus aspects (N) revealed apparent reduced and left relocated heterochromatin and underlined the outer inner membrane of envelope where examples of pore defects are noted (narrow arrows). One nucleus (thick white arrow) revealed an apoptotic-like stuffed nucleus with patches of obvious contrast heterochromatin among other ribonucleoproteins. Several injured cells showed most organelles dilated, endoplasm (ser) with RER with local patched, jammed ribonucleoprotein remains (see insert). Adjacent to nuclei, some rare mitochondria profiles (mt, black arrows) and huge phagosomes; n: nucleolus; ph: phagosome.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Membrane

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 9. a-n: TEM of a series of DU145 cell nuclei. a: control untreated cell nucleus (with enlarged nucleolus an) to compare with b-l aspects out of any combined VK3:VC treatment as they either underwent steps involving chromatin degradation by chromatolysis (b to h) and associated nucleoli where hetero chromatin (dense fibrillar part) left the fine fibrillar and the granular components as those parts got compacted together, shrunk and fragment; with remained heterochromatin as aggre gate grains and contrasted patches within the nucleoplasm (curved arrows), and alongside the inner nuclear membrane. Alternatively, other examples showed (i to m), as apoptotic- like chromatin pattern (i) followed by filled to morcellated nucleoplasm, and adjacent domains degenerated. Nucleoplasm chromatin degraded can appear as electron dense pond-like surrounded by vacuoles swellings out of misbuilt envelope that participated to the organelle degeneration. At the end, there is a complete nucleus structure annihilation (n).

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Control, Membrane

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 10. DU145 cell located in the upper region of oral VK3:VC treated mice that exemplified one a large cytoplasm field enriched by ribonucleoproteins that demonstrated dilations of mitochondria inner cristae and matrix deposition of electron contrasted material amid interactive, swollen RER components and a few dispersed lipid droplets (li), also shown with similar in vitro treatment.70,71,75

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: In Vitro

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 11. a-d: TEM of a series mitochondria profiles found among treated DU145 cells. a: control untreated cell aspect that compared with b-d views observed out of any combined VK3:VC treatment. The micrograph set of sequential damages included intra matrix deposits, disappearance of cristae, and swelling toward ultimate envelope rupture with content remained as tiny debris; li: lipid droplets; mt: mitochondria’ N; nucleus; ph: phagosome; arrows mark ribonucleoproteins.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Control

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 13. a-d: LM (a-c) and TEM (d) of DU145 carcinoma from oral VK3:VC-treated mice. LM again emphasized the pleiomorphic- organized tumor tissue and revealed poorly intercellular connections between blocked cells at replication events (open arrows in a (telophase) and in b (prophase, metaphase, and telophase). Phagosomes (ph) and haphazard dysmorphology throughout the randomly cut fields of view suggested cell’s extensions (*) that underwent self-excisions (small arrows). d: example of stretched minute cell attachments or junctions; c: connective components (cells and ground substance); Li: lipid droplets; N: nucleus; v: vascular structures.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques:

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 14. a-c: TEM aspects of DU145 carcinoma tissue after mice had received oral treatment with VK3: VC combination. Pleiomorphic-organized tumor tissue again showed diverse autolytic damages revealed by nucleus profile changes, including those with “smeared” chromatin (a), nucleolus disappearance, swelled organelles with fused damage (a), and electron dense phagosomes (a and c). Insert of a (curved arrow) marked lysosomes associated with ongoing cytophagocytosis noted by distorted adjacent cells that formed a huge phagosome. Open arrow in B indicated a deeply indented nucleus while an adjacent, worse injured cell split.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques:

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 15. a-f: LM (a-d) and TEM (e-f) devastation aspects of DU145 carcinoma and cells after mice had received both oral and IP VK3: VC combination treatment. The overall dismantled aspect of a in luminal space (l) completed by the shown awkward epithelial surface (b), cytoplasm self-excisions (a, b small arrows, star in e). In a: a small area of malignant island surrounded nests of malignant cells, enlarged in d, showing with toluidine blue stain pattern at least 3 cell types (e: eosinophil; b: basophil, i: intermediate). In c: open arrows mark autophagocytosis; in E: nucleolus damage and internal swellings or endomembrane damages throughout all cells (e and f). In e and f, escaped erythrocytes from local damaged vessels among DU145 cells (stars).

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Staining

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 16. a-f: TEM aspects from one mouse DU145 gauged xenograft after treatment by oral or IP VK3: VC combination. The series of DU145 carcinoma cells showed disjoining and freed cells with high nuclear: cytoplasm ratio. Still attached cells (a and e-f) undergo apical swelling (*) and freed cells (b-d) drifted, engaged in removing hung and ballooned cytoplasm parts (*) that characterized autoschizis.52,59,75,90 in F, the magnified featured excision part of E involved cytoskeleton bands (small arrows) that barred the perinuclear cell region. However, within the ballooned excising region, fibrillar bits or segments of cytoskeleton (small arrows) can be seen without evident organelle. L: lumen; arrowed mt: mitochondria in insert of E; white star: erythrocyte.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques:

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 17. a-e: LM (a, b, d, e) and TEM (c) featured a dismantled region part of one xenografted DU145 tumor growing on the diaphragm of a nude mouse treated by both oral and IP VC: VK3 combination. Evident eroded and rift parts showed in a and b where luminal (l) aspect showed freed cells, corpses, or debris, with few erythrocytes (open arrows). Injured carcinoma cells detached revealed damaged nucleus (n) surrounded by rims of cytoplasm. Detached and injured carcinoma bulk tissue showed cell’s autoschizis aka self-excisions caused by protruding appendages extended away from the “cell bodies” (nucleus and cytoplasm) that resemble those found in vitro after treatment with VK3: VC in vitro.52,54,59,70,71,75,90–92 in b (LM) and c (TEM) featured extremely damaged cells, as in Figure 15(e-f), containing endomembrane damages, including a large central pool of lytic proteins (poorly basophilic in b) smeared, adjacent to a Golgi apparatus parts, while the karyolitic nucleus (n) contained nucleoli parts (n). d and e illustrated buddings while karyolitic nucleus preceded a “liquefaction” necrosis of the DU145 cells (black arrow in e).

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: In Vitro

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 18. a-b: diagrams to attempt illustrating DU145 cell nucleus that bore cuts of DNA (heterochromatin filaments) as attached to inner membrane from: a: interphase cell where DNA, supplied in NOR of nucleolus (n) became recoiled parts at inner membrane attachment sites of envelope and cut coiled parts appeared as nucleoplasm grouped dots in ultrathin sections. b: G2M or S phase nucleus that resulted in amassed replicated DNA oligomers with other lysed compounds in a nucleoplasm reconstructed with damaged envelope. Open arrowheads: DNase I, and black arrow heads : DNase II. Figure 18 a-g Supplement: Histochemical detec tion of nucleases by Gomori modified technique in rat hepatoma induced by nitrosamine. Frozen section counterstained by eosin. In a and d: Control tissue with no nuclease activity induction. b: Alkaline DNase revealed with menadione bisulfite at the level of the nucleus envelopes and in cytoplasm (mitochondria). In c: Alkaline RNase can be seen localized in nucleoli (arrows). e-g : Nucleases after menadione and ascorbate stimulus; e: Acid RNase in cytoplasm; f and g: Acid DNase revealed in nucleus envelope and content,; note the enzyme nucleus locations is especially associated with the frequent replicated cell chromo somes (white arrows). Obtained out of collection of Taper’s micrographs after 187,189 and 193 published manuscripts were prepared.

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Membrane, Modification, Control, Activity Assay

Journal: Ultrastructural pathology

Article Title: Reactivation of nucleases with peroxidation damages induced by a menadione: ascorbate combination devastates human prostate carcinomas: ultrastructural aspects.

doi: 10.1080/01913123.2024.2379300

Figure Lengend Snippet: Figure 19. Cell structure damaged and associated time lapses to show highest alkaline and acid nuclease histochemical activities from each vitamin treatment as adapted from29 that agreed also with our fine structure data. Figure 19 Supplement: Pane illustrating the reactivation of DNases in excised implanted DU145 human prostate tumor on diaphragm of nude mice after one single-dose administration of IP and oral vitamins C and K3 (or Group III). Left column: Alkaline DNase (DNase I); little. or no alkaline DNase activity found in the Sham-treated tumor (C or Group IV). However, there is distinct alkaline DNase activity primarily in the nuclei 1hour after injection. This activity decreased during the second hour (2 h) and became difficult to appreciate at later time points (4 h, 8 h, 24 h). Right column: Acid DNase; DNase II activity observed in the tumor after the same vitamin treatment. Little or no activity observed in the Sham-treated tumor (C or Group IV). Progressive activation of DNase II occurred over 2 hr after vitamin treatment, and DNase II activity was at a maximum after 4 h post IP. This activity decreases after 8 h and is still evident after 24 h. Scale bar is 20 µm. Copy of Figure 1 (page 112), from Taper HS, Jamison JM, Gilloteaux J, Gwin CA, Gordon T, Summers JL. In Vivo Reactivation of DNases in Implanted Human Prostate Tumors After Administration of a Vitamin C/K3 Combination. Journal of Histochemistry & Cytochemistry. 2001;49(1):109-119. doi: 10.1177/002215540104900111 (with permission of Sage Publications India Pvt Ltd 248,001 Uttarakhand, India).

Article Snippet: DU145 human androgen-independent prostate carcinoma cell line, obtained from the American Type Culture Collection (Rockville, MD) and were grown in a humidified atmosphere at 37°C in 5% CO2 in McCoy’s 5A medium (M5A, Gibco, Grand Island, NY) and supplemented with 10% fetal bovine serum (FBS; Gibco; 2.2 g/L NaHCO3 and 50 μg/mL gentamicin sulfate (Fisher Scientific, Pittsburgh, PA).

Techniques: Activity Assay, Injection, Activation Assay, In Vivo

Journal: Food Bioscience

Article Title: Assessment of the in-vitro toxicity and in-vivo therapeutic capabilities of Juglans regia on human prostate cancer and prostatic hyperplasia in rats

doi: 10.1016/j.fbio.2023.103539

Figure Lengend Snippet: Fig. 3. Real-time, label-free, and high-throughput monitoring of Juglans regia fresh fruits extract effects on DU145 prostate carcinoma and PNT1A normal prostate human cells with iCELLigence system. The yellow arrow indicates the treatment point (~24th hour). A. DU145 cells B. PNT1A cells. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: DU145 human prostate carcinoma cell line (Catalogue no: HTB-81) was purchased from American Type Culture Collection (ATCC, Maryland, USA).

Techniques: High Throughput Screening Assay