Chemicals and Drugs Commons^™

Cross-Sector Review Of Drivers And Available 3rs Approaches For Acute Systemic Toxicity Testing, Troy Seidle, Sally Robinson, Tom Holmes, Stuart Creton, Pilar Prieto, Julia Scheel, Magda Chlebus

Troy Seidle, PhD

Acute systemic toxicity studies are carried out in many sectors in which synthetic chemicals are manufactured or used and are among the most criticized of all toxicology tests on both scientific and ethical grounds. A review of the drivers for acute toxicity testing within the pharmaceutical industry led to a paradigm shift whereby in vivo acute toxicity data are no longer routinely required in advance of human clinical trials. Based on this experience, the following review was undertaken to identify (1) regulatory and scientific drivers for acute toxicity testing in other industrial sectors, (2) activities aimed at replacing, reducing, or …

A Thousand Tiny Pieces: The Federal Circuit’S Fractured Myriad Ruling, Lessons To Be Learned, And The Way Forward, Jonathan R. K. Stroud

Jonathan R. K. Stroud

No abstract provided.

Erectogenic Effects Of Clerodendron Capitatum: Involvement Of Phosphodiesterase Type-5 Inhibition, Dr Syam Mohan

Syam Mohan

No abstract provided.

Clausena Excavata Burm. F. (Rutaceae): A Review Of Its Traditional Uses, Pharmacological And Phytochemical Properties., Dr Syam Mohan

Syam Mohan

The present review describes the morphological, traditional, phytochemical and pharmacological aspects of Clausena excavata (Burm. f., Rutaceae). The plant grows wild in the tropical and subtropical parts of Asia and represents one of the richest sources of medicinal natural products. It is intensively used currently in traditional medicine for treatment of different illnesses. One part or a mixture of two or more parts of the plant in the form of powder or liquid doze is usually given to the patient. Phytochemically, many pure compounds from several secondary metabolite groups of C. excavata have been isolated from different parts of the …

Chloroform Fraction Of Centratherum Anthelminticum (L.) Seed Inhibits Tumor Necrosis Factor Alpha And Exhibits Pleotropic Bioactivities: Inhibitory Role In Human Tumor Cells, Dr Syam Mohan

Syam Mohan

We investigated the antioxidant potential, cytotoxic effect, and TNF-α inhibition activity with NF-κB activation response in a chloroformfraction of Centratherum anthelminticum seeds (CACF). The antioxidant property of CACF was evaluated with DPPH, ORAC, and FRAP assays, which demonstrated significant antioxidant activity. The cytotoxicity of CACF was tested using theMTT assay; CACF effective inhibitory concentrations (IC50) for A549, PC-3, MCF-7, and WRL-68 cells were 31.42 ± 5.4, 22.61 ± 1.7, 8.1±0.9, and 54.93±8.3 μg/mL, respectively. CACF effectively and dose-dependently inhibited TNF-α release, in vitro and in vivo. CACF inhibited TNF-α secretion in stimulated RAW264.7macrophage supernatants with an IC50 of 0.012 μg/mL, …

In Vitro Anti-Inflammatory, Cytotoxic And Antioxidant Activities Of Boesenbergin A, A Chalcone Isolated From Boesenbergia Rotunda (L.) (Fingerroot), Dr Syam Mohan

Syam Mohan

The current in vitro study was designed to investigate the anti-inflammatory, cytotoxic and antioxidant activities of boesenbergin A (BA), a chalcone derivative of known structure isolated from Boesenbergia rotunda. Human hepatocellular carcinoma (HepG2), colon adenocarcinoma (HT-29), non-small cell lung cancer (A549), prostate adenocarcinoma (PC3), and normal hepatic cells (WRL-68) were used to evaluate the cytotoxicity of BA using the MTT assay. The antioxidant activity of BA was assessed by the ORAC assay and compared to quercetin as a standard reference antioxidant. ORAC results are reported as the equivalent concentration of Trolox that produces the same level of antioxidant activity as …

Synthesis Of Chalcones With Anticancer Activities., Dr Syam Mohan

Syam Mohan

Several chalcones were synthesized and their in vitro cytotoxicity against various human cell lines, including human breast adenocarcinoma cell line MCF-7, human lung adenocarcinoma cell line A549, human prostate cancer cell line PC3, human adenocarcinoma cell line HT-29 (colorectal cancer) and human normal liver cell line WRL-68 was evaluated. Most of the compounds being active cytotoxic agents, four of them with minimal IC50 values were chosen and studied in detail with MCF-7 cells. The compounds 1, 5, 23, and 25 were capable in eliciting apoptosis in MCF-7 cells as shown by multiparameter cytotoxicity assay and caspase-3/7, -8, and -9 activities …

Cucurbitacin L 2-O- Glucoside Demonstrates Apoptogenesis In Colon Adenocarcinoma Cells (Ht-29): Involvement Of Reactive Oxygen And Nitrogen Species Regulation, Dr Syam Mohan

Syam Mohan

Emerging evidence suggests that reactive oxygen (ROS) and nitrogen (RNS) species can contribute to diverse signalling pathways of inflammatory and tumour cells. Cucurbitacins are a group of highly oxygenated triterpenes. Many plants used in folk medicine to treat cancer have been found to contain cucurbitacins displaying potentially important anti-inflammatory actions. The current study was designed to investigate the anti-ROS and -RNS effects of cucurbitacin L 2-O-β-glucoside (CLG) and the role of these signaling factors in the apoptogenic effects of CLG on human colon cancer cells (HT-29). This natural cucurbitacin was isolated purely from Citrullus lanatus var. citroides (Cucurbitaceae). The results …

Dentatin Induces Apoptosis In Prostate Cancer Cells Via Bcl-2, Bcl-Xl, Survivin Downregulation, Caspase-9, -3/7 Activation And Nf-Kb Inhibition., Dr Syam Mohan

Syam Mohan

This study was set to investigate antiproliferative potential of dentatin (a natural coumarin isolated from Clausena excavata Burm. F) against prostate cancer and to delineate the underlying mechanism of action. Treatment with dentatin dose-dependently inhibited cell growth of PC-3 and LNCaP prostate cancer cell lines, whereas it showed less cytotoxic effects on normal prostate epithelial cell line (RWPE-1). The inhibitory effect of dentatin on prostate cancer cell growth was due to induction of apoptosis as evidenced by Annexin V staining and cell shrinkage. We found that dentatin-mediated accumulation of reactive oxygen species (ROS) and downregulated expression levels of antiapoptotic molecules …

The Growth Suppressing Effects Of Girinimbine On Hepg2 Involve Induction Of Apoptosis And Cell Cycle Arrest, Syam Mohan

Syam Mohan

Murraya koenigii is an edible herb widely used in folk medicine. Here we report that girinimbine, a carbazole alkaloid isolated from this plant, inhibited the growth and induced apoptosis in human hepatocellular carcinoma, HepG2 cells. The MTT and LDH assay results showed that girinimbine decreased cell viability and increased cytotoxicity in a dose-and time-dependent manner selectively. Girinimbine-treated HepG2 cells showed typical morphological features of apoptosis, as observed from normal inverted microscopy and Hoechst 33342 assay. Furthermore, girinimbine treatment resulted in DNA fragmentation and elevated levels of caspase-3 in HepG2 cells. Girinimbine treatment also displayed a time-dependent accumulation of the Sub-G(0)/G(1) …

Ppar Agonists Down-Regulate The Expression Of Atp10c Mrna During Adipogenesis, A Peretich, Maria Cekanova Ms, Rndr, Phd, S Hurst, Sj Baek, Madhu Dahr

Maria Cekanova MS, RNDr, PhD

No abstract provided.

Effects Of Angiotensin 1-7 On The Actions Of Angiotensin Ii In The Renal And Mesenteric Vasculature Of Hypertensive And Streptozotocin-Induced Diabetic Rats, Mohd Rais Mustafa, Murugan Dharmani, Francis I. Achike, Meng-Kwoon Sim

Mohd Rais Mustafa

Angiotensin 1-7, a heptapeptide derived from metabolism of either angiotensin I or angiotensin II, is a biologically active peptide of the renin–angiotensin system. The present study investigated the effect of angiotensin 1-7 on the vasopressor action of angiotensin II in the renal and mesenteric vasculature of Wistar-Kyoto (WKY) rats, spontaneously hypertensive rats (SHR) and streptozotocin-induced diabetic rats. Angiotensin II-induced dose-dependent vasoconstrictions in the renal vasculature. The pressor response was enhanced in the SHR and reduced in the streptozotocin-diabetic rat compared to WKY rats. Angiotensin 1-7 attenuated the angiotensin II pressor responses in the renal vasculature of WKY and SHR rats. …

Modulation Of Vascular Reactivity In Normal, Hypertensive And Diabetic Rat Aortae By A Non-Antioxidant Flavonoid, Mohd Rais Mustafa, Macha Ajay, Fracis I. Achike

Mohd Rais Mustafa

In this study, we report the effects of a non-antioxidant flavonoid flavone on vascular reactivity in Wistar–Kyoto (WKY) rat isolated aortae. Whether flavone directly modulates vascular reactivity in spontaneously hypertensive rat (SHR) and streptozotocin-induced diabetic-WKY rat isolated aortae was also determined. Thoracic aortic rings were mounted in organ chambers and exposed to various drug treatments in the presence of flavone or its vehicle (DMSO), which served as control. Pretreatment with flavone enhanced relaxant effects to endothelium-dependent vasodilator acetylcholine (ACh) and attenuated contractile effects to 1-receptor agonist phenylephrine (PE) in WKY aortae compared to those observed in control aortic rings. Flavone …

Effects Of Ascorbic Acid On Impaired Vascular Reactivity In Aortas Isolated From Age-Matched Hypertensive And Diabetic Rats, Mohd Rais Mustafa, Macha Ajay

Mohd Rais Mustafa

Impaired vascular reactivity is a hallmark of several cardiovascular diseases that include hypertension and diabetes. This study compared the changes in vascular reactivity in age-matched experimental hypertension and diabetes, and, subsequently, tested whether these changes could be affected directly by ascorbic acid (10 microM). Endothelium-derived nitric oxide (NO) modulation of ascorbic acid effects was also investigated. All the experiments were performed in the presence of a cyclooxygenase inhibitor, indomethacin (10 microM). Results showed that the endothelium-dependent and -independent relaxations induced by acetylcholine (ACh) and sodium nitroprusside (SNP), respectively, were blunted to a similar extent in isolated aortic rings from age-matched …

Bioassay-Guided Isolation Of A Vasorelaxant Active Compound From Kaempferia Galanga L, Mohd Rais Mustafa, Khalijah Awang, Mustafa Ali Mohd, Rozana Othman, Halijah Ibrahim

Mohd Rais Mustafa

Bioassay-guided fractionation was performed on a crude dichloromethane extract of Kaempferia galanga L. using chromatography techniques. Screening of the extract for biological activity started with the brine shrimp lethality bioassay, followed by the study of its antihypertensive activity on anaesthetized rats, which involved monitoring of the extract's effect on mean arterial blood pressure. The components of the fractions obtained from the separation procedures were analyzed using gas chromatography (GC). The yield of the CH2Cl2 extract was 0.29% of the crude plant extract. Analysis of the data for brine shrimp lethality test using the Finney computer program showed that this extract …

Expression Of G-Protein Inwardly Rectifying Potassium Channels (Girks) In Lung Cancer Cell Lines, Howard Plummer 3rd, Madhu Dhar, Maria Cekanova Ms, Rndr, Phd, Hildegard Schuller

Maria Cekanova MS, RNDr, PhD

BACKGROUND: Previous data from our laboratory has indicated that there is a functional link between the beta-adrenergic receptor signaling pathway and the G-protein inwardly rectifying potassium channel (GIRK1) in human breast cancer cell lines. We wanted to determine if GIRK channels were expressed in lung cancers and if a similar link exists in lung cancer. METHODS: GIRK1-4 expression and levels were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. GIRK protein levels were determined by western blots and cell proliferation was determined by a 5-bromo-2'-deoxyuridine (BrdU) assay. RESULTS: GIRK1 mRNA was expressed in three of six small …

Effects Of Des-Aspartate-Angiotensin I On The Actions Of Angiotensin Iii In The Renal And Mesenteric Vasculature Of Normo-And Hypertensive Rats, Mohd Rais Mustafa, Murugan Dharmani, Kunheen N.K, Meng-Kwoon Sim

Mohd Rais Mustafa

An earlier study showed that des-aspartate-angiotensin I (DAA-I) attenuated the pressor action of angiotensin III in aortic rings of the spontaneously hypertensive rat (SHR) but not the normotensive Wistar Kyoto (WKY) rat. The present study investigated similar properties of DAA-I in isolated perfused kidneys and mesenteric beds of WKY and SHR. In the renal vasculature, angiotensin III induced a dosedependent pressor response, which was more marked in the SHR than WKY in terms of significant greater magnitude of response and lower threshold. DAA-I attenuated the pressor action of angiotensin III in both the WKY and SHR. The attenuation in SHR …

Multiple Luteinizing Hormone Receptor (Lhr) Protein Variants, Interspecies Reactivity Of Anti-Lhr Mab Clone 3b5, Subcellular Localization Of Lhr In Human Placenta, Pelvic Floor And Brain, And Possible Role For Lhr In The Development Of Abnormal Pregnancy, Pelvic Floor Disorders And Alzheimer's Disease, A Bukovsky, K Indrapichate, H Fujiwara, Maria Cekanova Ms, Rndr, Phd, Me Ayala, R Dominguez, Mr Caudle, J Wimalsena, Rf Elder, P Copas, Jf Foster, Ri Fernando, Dc Henley, Nb Upadhyaya

Maria Cekanova MS, RNDr, PhD

Distinct luteinizing hormone receptor (LHR) protein variants exist due to the posttranslational modifications. Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages. The 3B5 mouse monoclonal antibody was raised against purified rat LHR. In rat, porcine and human ovaries, the 3B5 identified six distinct LHR bands migrating at approximately 92, 80, 68, 59, 52 and 48 kDa. Characteristic LHRI was detected in rat, human and porcine corpora lutea. During cellular differentiation, subcellular LHR distribution changed from none to granular cytoplasmic, perinuclear, surface, nuclear and no staining. …

Expression And Localization Of Estrogen Receptor-Alpha Protein In Normal And Abnormal Term Placentae And Stimulation Of Trophoblast Differentiation By Estradiol, A Bukovsky, Maria Cekanova Ms, Rndr, Phd, Mr Caudle, J Wimalasena, Js Foster, Dc Henley, Rf Elder

Maria Cekanova MS, RNDr, PhD

Estrogens play an important role in the regulation of placental function, and 17-beta-estradiol (E2) production rises eighty fold during human pregnancy. Although term placenta has been found to specifically bind estrogens, cellular localization of estrogen receptor alpha (ER-alpha) in trophoblast remains unclear. We used western blot analysis and immunohistochemistry with h-151 and ID5 monoclonal antibodies to determine the expression and cellular localization of ER-alpha protein in human placentae and cultured trophoblast cells. Western blot analysis revealed a ~65 kDa ER-alpha band in MCF-7 breast carcinoma cells (positive control). A similar band was detected in five normal term placentae exhibiting strong …

Placental Expression Of Estrogen Receptor Beta And Its Hormone Binding Variant – Comparison With Estrogen Receptor Alpha And A Role For Estrogen Receptors In Asymmetric Division And Differentiation Of Estrogen-Dependent Cells, Antonin Bukovsky, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Romaine I. Fernando, Jay Wimalasena, James S. Foster, Donald C. Henley, Robert F. Elder

Maria Cekanova MS, RNDr, PhD

During human pregnancy, the production of 17-beta-estradiol (E2) rises steadily to eighty fold at term, and placenta has been found to specifically bind estrogens. We have recently demonstrated the expression of estrogen receptor alpha (ER-alpha) protein in human placenta and its localization in villous cytotrophoblast (CT), vascular pericytes, and amniotic fibroblasts. In vitro, E2 stimulated development of large syncytiotrophoblast (ST) aggregates. In the present study we utilized ER-beta affinity purified polyclonal (N19:sc6820) and ER-alpha monoclonal (clone h-151) antibodies. Western blot analysis revealed a single ~52 kDa ER-beta band in chorionic villi (CV) protein extracts. In CV, strong cytoplasmic ER-beta immunoreactivity …

Variability Of Placental Expression Of Cyclin E Low Molecular Weight Variants, A Bukovsky, Maria Cekanova Ms, Rndr, Phd, Mr Caudle, J Wimalasena, Js Foster, Ja Keenan, Rf Elder

Maria Cekanova MS, RNDr, PhD

No abstract provided.

Vasorelaxant Effects Of Ethyl Cinnamate Isolated From Kaempferia Galanga On Smooth Muscles Of The Rat Aorta, Mohd Rais Mustafa, Rozana Othman, Halijah Ibrahim, Mustafa Ali Mohd, Khalijah Awang, Anwar-Ul Hassan Gilani

Mohd Rais Mustafa

From the rhizomes of Kaempferia galanga, ethyl cinnamate (EC) was isolated and its vasorelaxant effect was examined on the rat aorta. EC inhibited the tonic contractions induced by high K+ and phenylephrine (PE) in a concentration-dependent manner, with respective IC50 values of 0.30 ± 0.05 mM and 0.38 ± 0.04 mM. The relaxant effect against PE-induced contractions was greater in the presence of endothelium. Pre-treatment of the aorta with methylene blue and indomethacin significantly reduced the relaxant effect. These results suggest that the inhibitory effects of EC may involve inhibition of Ca2+ influx into vascular cells and release of nitric …

Abnormal Expression Of P27kip1 Protein In Levator Ani Muscle Of Aging Women With Pelvic Floor Disorders – A Relationship To The Cellular Differentiation And Degeneration, Antonin Bukovsky, Pleas Copas, Michael R. Caudle, Maria Cekanova Ms, Rndr, Phd, Tamara Dassanayake, Bridgett Asbury, Stuart E. Van Meter, Robert F. Elder, Jeffrey B. Brown, Stephanie B. Cross

Maria Cekanova MS, RNDr, PhD

Background Pelvic floor disorders affect almost 50% of aging women. An important role in the pelvic floor support belongs to the levator ani muscle. The p27/kip1 (p27) protein, multifunctional cyclin-dependent kinase inhibitor, shows changing expression in differentiating skeletal muscle cells during development, and relatively high levels of p27 RNA were detected in the normal human skeletal muscles.

Methods Biopsy samples of levator ani muscle were obtained from 22 symptomatic patients with stress urinary incontinence, pelvic organ prolapse, and overlaps (age range 38–74), and nine asymptomatic women (age 31–49). Cryostat sections were investigated for p27 protein expression and type I (slow …

Oscillations And Waves Of Intracellular [Ca2+] In Cardiac-Muscle-Cells, Mohd Rais Mustafa

Mohd Rais Mustafa

When cardiac muscle cells become overloaded with Ca2+, spontaneous releases of Ca2+ from the sarcoplasmic reticulum can occur. Such spontaneous releases normally originate within a localized region of the cell and propagate as a wave throughout the cell. The amplitude and frequency of these Ca2+ releases increase with the degree of Ca2+ loading of the cell, and are abolished by interventions which inhibit sarcoplasmic reticulum function. It is still not clear whether these releases are triggered by a rise in the [Ca2+] in the lumen of the sarcoplasmic reticulum or by a rise in cytoplasmic [Ca2+]. Such releases may, however, …