E. I. Medine

Isolation of hydroxytyrosol from olive leaves extract, radioiodination and investigation of bioaffinity using in vivo/in vitro methods

Summary It is known that medicinal plants like olive have biological activities due to their flavonoid content such as olueropein, tyrosol, hydroxytyrosol etc. In current study, hydroxytrosol (HT) which is one of the major phenolic compounds in olive, olive leaves and olive oil, was isolated after methanol extraction and purification of olive leaves which are grown in the northern Anatolia region of Turkey. The isolated HT was radiolabeled with 131I (131I-HT) and the bioaffinity of this radiolabeled component of olive leaves extract was investigated by using in vivo/in vitro methods. It was found that HT could be radiolabeled with 131I in yields of 95.6±4.4% (n = 8), and in vivo studies showed that 131I-HT is taken up by urinary bladder, stomach, small intestine, large intestine, breast and prostate. Significant incorporation of activity was observed in cell lines via in vitro studies.

L-Carnitine Protection Against Cisplatin Nephrotoxicity In Rats: Comparison with Amifostin Using Quantitative Renal Tc 99m DMSA Uptake

Objective: In this study, we aimed to investigate the cytoprotective effect of L-carnitine against cisplatin-induced nephrotoxicity and to compare its efficacy with that of amifostin by quantitative renal Tc 99m DMSA uptake. Material and Methods: Male Wistar rats were randomly divided into six groups of six animals each. 1) Control (saline; 5 ml/kg intraperitoneally); 2) L-carnitine (CAR; 300 mg/kg intraperitoneally); 3) Amifostine (AMI; 200 mg /kg intraperitoneally); 4) Cisplatin (CIS;7 mg/kg intraperitoneally); 5) Cisplatin plus L-carnitine (CIS + CAR); 6) Cisplatin plus amifostine (CIS + AMI). L-carnitine and amifostine were injected 30 minutes before cisplatin in Group 5 and 6. Tc 99m DMSA, 7.4 MBq/0.2 ml, was injected through the tail vein 72 hours after the drug administration. Rats were killed and kidneys removed by dissection 2 hours after the injection of the radiopharmaceutical. The percentage of the injected dose per gram of kidney tissue (%ID/g) was calculated. Renal function was monitored by measuring BUN and plasma levels of creatinine. Lipid peroxidation and glutathione content were determined by measuring malondialdehyde (MDA) and reduced glutathione (GSH) in kidney tissue homogenates. Results: Tc 99m DMSA uptake per gram tissue of the kidney as %ID/g was 29.54±4.72, 29.86 ± 7.47 and 26.37 ± 4.54 in the control, CAR and AMI groups respectively. %ID/g was the lowest of all the groups, 11.60±3.59 (p<0.01), in the cisplatin group. Carnitine or amifostine administration 30 minutes before cisplatin injection resulted a significant increase in %ID/g, 21.28±7.73 and 18.97±3.24 respectively, compared to those of cisplatin-treated rats (p<0.002). A marked increase in plasma BUN and creatinine indicating nephrotoxicity and acute renal failure was observed in the cisplatin-treated group. MDA and GSH levels were concordant with cisplatin-induced oxidative stress in the kidney tissue. Conclusion: The results showed that L-carnitine significantly attenuates the cisplatin-induced nephrotoxicity as amifostin. Conflict of interest:None declared.

Radiolabeling of methanol extracts of yarrow (Achillea millefolium l) in rats

PURPOSE Current study is focused on extraction with methanol, purification, labeling with (131)I using iodogen method of the yarrow plant and investigating in vivo biological activity using biodistribution and imaging studies on healthy animal models. The aim of the study is to contribute plant extracts to discover new drugs in the diagnosis and treatment of several diseases. METHODS Nine female and nine male healthy Wistar albino rats, which were approximately 100-150 g in weight, were used for biodistribution studies. For imaging studies four healthy male Balb-C mice were used. Quality control studies were done utilizing thin layer radio chromatography (TLRC) and high performance liquid chromatography (HPLC) methods. For biodistribution studies, (131)I radiolabeled Peak 7 ((131)I-Peak 7) was sterilized and injected into the tail veil of rats and imaging studies were obtained using Kodak FX PRO in vivo Imaging System. RESULTS The radiolabeling yield of each purified the bioactive extracts of the yarrow plant, seven peaks was between 79 and 92%. The highest radiolabeling yield was calculated for (131)I radiolabeled seventh peak ((131)I-Peak 7) (92.78 ± 5.04, n=5). For this reason the biodistribution and imaging studies were done for (131)I-Peak 7. Thats why; these studies with Peak 7 were carried out. CONCLUSION Peak 7 was radiolabeled with (131)I in high yield for using imaging and therapeutic studies in nuclear medical applications.

Synthesis of a theranostic agent: radioiodinated PEGylated PLGA-indocyanine capsules and in vitro determination of their bioaffinity on ovarian, cervical and breast cancer cells

The aim of current study is to synthesize a theranostic (multi-functional) agent, which is targeted on ovary, cervical and breast cancer types with diagnosis and treatment potential and to determine its bioaffinity by using in vitro methods. In conclusion; the designed compound (IPPP), which has fluorescence capability (from Indocyanine), encapsulated structure (with PEGylated PLGA), included an anticancer drug (Paclitaxel) for targeting and radionuclidic tracer (131I) content for tracing, has bioaffinity and promise for diagnosis and therapy on ovarian, cervical and breast cancer cell lines.

Investigation of Therapeutic Efficiency of Bleomycin and Bleomycin-Glucuronide Labeled with 131I on the Cancer Cell Lines

The aim of this study is to determine the incorporations of radiolabeled bleomycin ((131)I-BLM) and bleomycin-glucuronide ((131)I-BLMGLU) on PC-3 (human prostate carcinoma cell line), Caco-2 (human colon adenocarcinoma cell line), Hutu-80 (Human Duodenum adenocarcinoma cell line), and A549 (Human lung adenocarcinoma epithelial cell line) cancerous cell lines. For this purpose, BLM and BLMGLU enyzmatically synthesized were labeled with (131)I, quality control studies were done and the incorporation yields of (131)I-BLM and (131)I-BLMGLU on these cell lines were measured. Quality-control studies showed that the radiolabeling yields were obtained as 95% and 90% for (131)I-BLM and (131)I-BLMGLU, respectively. Also, as a result of the cell culture studies, it was found that (131)I-BLM and (131)I-BLMGLU had higher incorporation on PC-3 cells than that of other cell lines. In addition to this, it was reported that the incorporation yield of (131)I-BLMGLU was higher than that (131)I-BLM. At the end of the study, cytotoxicities of BLM and BLMGLU on PC-3 cancerous cell line were inspected and fluorescent images of BLM and BLMGLU were taken on PC-3 cells by using fluorescein isothiocyanate. In conclusion, cell culture studies demonstrated that the incorporation values of (131)I-BLMGLU on the four cell lines were about five to six times higher than (131)I-BLM. Radiolabeled glucuronide derivatives can be used in cancer therapy and tumor imaging, depending on the properties of radioiodine for the β-glucuronidase-rich tissues because glucuronidation leads to rapid and higher incorporation on adenocarcinoma cells.

Synthesis, radiolabeling and biodistribution of morphine glucuronide (mor-glu)

The aim of this study was to synthesize a glucuronide conjugated morphine derivative which could be labeled with 131I, as a radiopharmaceutical, and to investigate its radiopharmaceutical potential using biodistribution studies in male Albino Wistar rats. Morphine was extracted from dry capsules of the opium poppy (Papaver somniferum L.). It was conjugated with UDP-glucuronic acid by using UDP-glucuronyl transferase (UDPGT) enzyme rich microsomes, purified by high performance liquid chromatography (HPLC) and characterized by nuclear magnetic resonance (NMR), infrared (IR) spectroscopy and liquid chromatography mass spectroscopy (LC-MS/MS). Normal and receptor blockage biodistribution studies were performed in male Albino Wistar rats. The results of the tissue distribution studies showed that 131I labeled morphine glucuronide (131I-mor-glu) uptake in the small intestine, large intestine and urinary bladder was higher than in the other tissues of the rats in the blocked receptor and unblocked receptor. A greater uptake of the radio labeled substance was observed in the hypothalamus and mid brain than in the other branches of the rats’ brains.

Extraction, radiolabeling, and biodistribution of thebaine in rats

The goal of this study was to determine the radiopharmaceutical potential of radioiodinated thebaine. Thebaine was extracted from dry capsules of opium poppy (Papaver somniferum L.), purified using high-performance liquid chromatography, and characterized with nuclear magnetic resonance and infrared spectroscopy. The purified thebaine was labeled with 131I using the iodogen method. Normal and receptor-blockage biodistribution studies were performed in male Albino Wistar rats. The results of the tissue distribution studies showed that the uptake of 131I-thebaine in the stomach, large intestine, spinal cord, and prostate was higher than in the other tissues. A greater uptake of radiolabeled thebaine in the rat brain was observed in the midbrain and hypothalamus. We concluded that (1) the labeling yield of 131I-thebaine was high, (2) a large amount of 131I-thebaine remained in the midbrain and hypothalamus, and (3) 131I-thebaine had enough stability for diagnostic scanning.

Radioiodination of cyclin dependent kinase inhibitor Olomoucine loaded Fe@Au nanoparticle and evaluation of the therapeutic efficacy on cancerous cells

Abstract Magnetic nanoparticles have promising biomedical applications such as drug delivery, novel therapeutics and diagnostic imaging. Magnetic drug delivery combination works on the delivery of magnetic nanoparticles loaded with drug to the target tissue by means of an external magnetic field. Gold coated iron oxide (Fe@Au) nanoparticles can provide useful surface chemistry and biological reactivity. Covalent conjugation to the Fe@Au nanoparticles through cleavable linkages can be used to deliver drugs to tumor cells, then the drug can be released by an external. In this paper, purine based cyclin dependent kinases (CDKs) inhibitor Olomoucine (Olo) [2-(Hydroxyethylamino)-6-benzylamino-9-methylpurine] was loaded on gold coated iron oxide (Fe@Au) nanoparticles and radiolabeled with 131I to combine magnetic targeted drug delivery and radiotherapy. Fe@Au nanoparticles were synthesized by microemulsion method. The characterization of nanoparticles was examined by TEM, VSM and XRD. Amine activation was utilized by cysteamine hydrochloride and then CDI was used for conjugation of Olomoucine. Antiproliferative effect and cytotoxicity of Olomoucine loaded Fe@Au nanoparticles (Fe@Au-Olo) were investigated on MCF7 and A549 cell lines. Proliferation rate was decreased while uptake of Fe@Au-Olo on both cell lines was high in comparison with Olomoucine. Also, enhanced incorporation ratio was observed under external magnetic field.

Multifunctional molecular imaging probes for estrogen receptors: 99mTc labeled diethylstilbestrol (DES) conjugated, cuinp quantum dot nanoparticles (DESCIP)

A theranostic nanoparticle was synthesized based on diethylstilbestrol conjugated with phosphate, copper, and indium (DESCIP) and labelled with 99mTc which can be used for SPECT imaging of ER-enriched cancers. In vitro biological activity of 99mTc-DESCIP was examined in breast adenocarcinoma cells (MCF-7), prostatic carcinoma cells (PC-3), and pulmonary epithelial cells (A-549). In vivo lymph node imaging was performed in normal and receptor blocked female New Zealand rabbits. Results demonstrated that 99mTc-DESCIP and DESCIP has potential for imaging ER-enriched tumors such as breast and prostate tumors, and their metastases in the lung, as well as improving management for their therapies.

INVESTIGATION OF 99MTC-LABELED CLIOQUINOL DERIVATIVE ON AMYLOID PLAQUE SPECIFICITY BY USING AN ANIMAL MODEL OF ALZHEIMER'S DISEASE

Objective: Alzheimer’s disease (AD), which comes up an important problem due to the aging population, has two hallmarks defined as amyloid plaques and neurofibrillary tangles. Despite the fact that AD affects a wide audience adversely, definitive diagnosis of AD still requires postmortem examination of the brain through histological staining of these hallmarks. Nowadays, these two hallmarks are playing crucial role in researches for diagnosis and treatment of AD. In addition, deficiency of AD diagnosis agents and limitation of assessing the blood-brain barrier (BBB) are main reasons of the current researches focusing on brain agents with plaque or tangle imaging potential. Metal-protein attenuating compounds (MPACs) such as clioquinol and cloxyquin have a great increasing promise on amyloid plaques. Yurt Kilcar et al. radiolabeled Bioquin-Hexa methyl propylene amine oxime (HMPAO) with 99mTc (99mTc-Bioquin-HMPAO) and evaluated the biodistribution on healthy male Balb/C mice (unpublished data). In current study, it is aimed to investigate amyloid plaque specifity of the 99mTc-BioquinHMPAO by using biodistribution studies on animal model of AD. Methods: Bioquin-HMPAO was labeled with 99mTc according to previous study. In accordance with the objectives of the study, animal model of AD created on healthy male Sprague Dawley rats with intrahippocampal stereotaxic injection of amyloid beta 1-42 (Aβ1-42). Intrahippocampal administration was applied to the CA1 area of hippocampus bilaterally (Aβ142 to the left side and PBS to the right side as control). Additionally, healthy male Sprague Dawley rats were utilized as naive group. Twenty days after surgery, biodistribution studies of 99mTc-Bioquin-HMPAO on animal model of AD and control group Susanne Granegger1, Lindsay Brammen2, Helmut Sinzinger1 1Institute ATHOS, Atherosclerosis Research, Vienna, Austria 2Vienna Medical University, Department of Nuclear Medicine, Vienna, Austria Elevated Lipoprotein(A) Impairs Platelet Radiolabeling Yield