Novel human interferon alpha 2b (hIFN<i>α</i>2b) muteins were developed by substituting cysteine residue (C) at positions 2 and 99 with aspartic acid residues (D). The mutein forms were then studied for pharmacokinetic profile. In addition, the influence of charge on the protein structure was tested <i>in vivo</i> for the biodistribution pattern. Codon substitutions were performed by Polymerase Chain Reaction (PCR)-based site-directed mutagenesis on a previously constructed synthetic hIFN<i>α</i>2b open reading frame (ORF) cloned in pET32b expression plasmid. The result of nucleotide sequencing analysis confirmed that all codons were replaced successfully without any additional mutation. Three mutant forms of hIFN<i>α</i>2b ORF were overexpressed in <i>Escherichia coli</i> BL21 (DE3) resulted in three muteins: hIFN<i>α</i>2b C2D, hIFN<i>α</i>2b C99D, hIFN<i>α</i>2b C2D C99D. To follow the kinetic and localization of the mutein interferon after intravenous administration, Tc^99m was used to label the proteins. In particular of elimination half-life, it was shown that hIFN<i>α</i>2b C2D C99D > hIFN<i>α</i>2bC2D > hIFN<i>α</i>2bC99D > wild type. hIFN<i>α</i>2b C2D C99D mutein showed highest blood accumulation after 30 minutes administration. Taken together, the charge of hIFN<i>α</i>2b seems to be responsible for the fate of hIFN<i>α</i>2b <i>in vivo</i>

Technetium-99m Methylene Diphosphonate (99mTc-MDP) has been utilized in a variety of clinical situations to identify bone areas due to the strong affinity of hydroxyapatite crystals in the mineral phase of the bone with the diphosphonate compounds. Osteoporosis is a disease characterized by decreased bone mass and increased fracture risk and represents a significant population health issue. It has been observed that 99mTc-MDP can be used for bone scintigraphy especially in case of bone cancer, but biodistribution study of 99mTc-MDP on ovariectomized mice for early monitoring of osteoporosis model remains unclear. Therefore, we aimed to investigate the biodistribution of 99mTc-MDP both in normal and ovariectomized mice. The experiment was performed on BALB/c mice weighing approximately 30 g. Mice were divided into a normal and ovariectomized group. After the first, second and third hours, mice were euthanized using the accepted protocol and the tissue of interest was collected. All tissue and blood were weighed using an analytical scale and counted for radioactivity using Automatic Gamma Counter with NaI(Tl) detector. Administration of 99mTc-MDP showed in normal mice compared with an animal model of osteoporosis, there are significant differences at 1 hour post-injection from (20.32±1.38) %ID/g decreased to (7.42±2.61) %ID/g, 2 hours from (13.75±0.01) %ID/g to (5.25±0.25) %ID/g and 3 hours from (12.18±1.44)%ID/g to (4.86±1.34) %ID/g uptake in the bones with (p<0.05). This study can be a consideration for the clinical application of 99mTc-MDP for early detection of osteoporosis conditions by looking at bone uptake and become a concern in the application for bone scintigraphy if the patient is indicated osteoporosis because it will affect visualization of the organ.

Mycobacterium tuberculosis merupakan bakteri tahan asam penyebab penyakit tuberkulosis (TB). Bakteri ini menyerang paru dan organ lain, seperti tulang, kulit, kelenjar getah bening, kelenjar tiroid, dan saluran urogenital. M. tuberculosis sangat mudah resisten terhadap obat anti tuberkulosis, sehingga berdampak pada sulitnya pengobatan penyakit TB secara tuntas. Resistensi yang terjadi umumnya diakibatkan oleh adanya mutasi. Deteksi resistensi M. tuberculosis terhadap INH dengan menggunakan teknik PCR (Polymerase Chain Reaction) untuk mengetahui adanya mutasi gen katG M. tuberculosis telah dilakukan pada penelitian sebelumnya. Kali ini dilakukan uji resistensi Mycobacterium tuberculosis terhadap kombinasi INH dan etambutol dengan menggunakan senyawa bertanda 99m Tc-etambutol secara in-vitro. Pengujian resistensi M. tuberculosis meliputi pemberian kombinasi obat INH dan etambutol pada M. tuberculosis, inkubasi, penambahan kadar antibiotik INH di minggu ke-2 pengamatan, penambahan senyawa bertanda 99m Tc-etambutol, dan pencacahan radioaktivitas. INH yang ditambahkan ke dalam kelompok tabung adalah 1 μg/mL, sedangkan kadar etambutol yang ditambahkan ke dalam kelompok tabung adalah 2 μg/mL, 4 μg/mL, 6 μg/mL, dan tanpa penambahan etambutol. Hasil penelitian menunjukkan bahwa M. tuberculosis yang telah diinduksi oleh INH dan etambutol masih dapat tumbuh secara bertahap selama 4 minggu inkubasi, sedangkan % uptake radioaktivitas 99m Tc-etambutol dan 99m Tc-perteknetat cenderung menurun dengan bertambahnya konsentrasi INH dan etambutol.

<p>Diabetes Melitus (DM) merupakan penyakit metabolik yang ditandai dengan tingginya kadar glukosa dalam darah (hiperglikemia). Penelitian ini bertujuan untuk mengetahui aktivitas ekstrak etanol herba suruhan sebagai antihiperglikemia dan peningkatan sensitivitas insulin. Hewan uji yang digunakan adalah tikus Wistar jantan yang dibagi ke dalam 6 kelompok (n=5), terdiri dari kelompok kontrol tikus normal, kontrol negatif, kontrol positif (metformin 45 mg/kg BB tikus), dan 3 kelompok dosis ekstrak etanol herba suruhan 50, 100, dan 150 mg/kgBB. Induksi hewan model hiperglikemia dilakukan dengan pemberian pakan tinggi lemak dan karbohidrat pada masing-masing kelompok selama 60 hari, kecuali kelompok kontrol tikus normal. Semua kelompok hewan dipuasakan selama 18 jam, kemudian kadar gula darahnya diukur. Pada hari ke 61 pengujian dilakukan pada masing-masing kelompok yang sudah diberi sediaan uji secara oral selama 7 hari, setelah itu kadar gula darah puasa akhir tikus diukur. Tes toleransi insulin dilakukan pada hari ke 71 setelah perlakuan. Hasil uji dengan membandingkan nilai konstanta tes toleransi insulin (KTTI) kelompok perlakuan terhadap kelompok tikus resistensi insulin menunjukkan terjadinya penurunan kadar glukosa darah dan peningkatan sensitivitas insulin yang signifikan (p≤0,05). Nilai KTTI pada ketiga kelompok dosis ekstrak etanol herba suruhan diperoleh berturut-turut sebesar 95,87; 74,44; dan 84,77 lebih besar dibandingkan dengan kelompok kontrol tikus resistensi insulin.</p>

Piperine, as one of the secondary metabolites of pepper, shows many pharmacological activities in various studies, including as antiinflammatory, antimicrobial, hepatoprotective, antioxidant, and potentially anticancer agents. Cancer is still one of the leading causes of death in the world due to the absence of specific symptoms in early-stage of cancer. The development of drugs for early diagnosis and selective treatment of cancer cells is considered one of the best solutions to reduce mortality. The activity of piperine that could induce apoptosis of 4T1 breast cancer cells and HT-29 colon cancer cells at in vivo trials prove it as a potential compound that can carry radioactive atoms (as labeledcompound) to cancer cells. This study was conducted to determine the optimal conditions of labeling of piperine with iodine-131 in order to obtain a 131 I-piperine which can later be used as a radiotracer in the development of cancer drugs. The labeling of piperine was performed by Chloramine-T iodination method and followed by purification with ion exchange chromatography. The optimal labeling results of piperine with 43.99% ± 1.23 radiochemical purity were obtained with piperine 3mg/300μL, chloramine-T 50 μg as an oxidizer, incubation time 30 min at room temperature, and 100 μg sodium metabisulphite as a reductor. To improve the purity of the preparation, purification by ion exchange chromatography method with Dowex 1X8 mesh 100-200 as resin was used until obtained 93.26% ± 0.94 of radiochemical purity. Further research is necessary before 131 I-piperine can be used as a radiotracer for cancer diagnosis in nuclear medicine.