Development of medium throughput muscarinic receptor binding assay

Muscarinic receptors are members of the large family of G protein-coupled receptors (GPCRs). These receptors are widely distributed throughout the body and are responsible for numerous vital functions in both the brain and autonomic nervous system (Lefkowitz et al., 1990)

Improved plasma stability and sustained release profile of gemcitabine via polypeptide conjugation

To enhance the stability of the anticancer drug gemcitabine (2′-deoxy-2′,2′-difluorocytidine), it was conjugated to poly-l-glutamic acid (PG-H) via a carbodiimide reaction. The synthesised poly-l-glutamic acid-gemcitabine (PG-G) was purified and characterised by using SDS-PAGE to estimate its molecular weight, HPLC to determine its purity and degree of drug loading, and NMR to elucidate the structure. In vitro aqueous hydrolytic studies showed that the gemcitabine release from the polymeric drug conjugate was pH dependent, and that the conjugation to PG-H improved its stability in human plasma. The release of the bound gemcitabine from PG-G in plasma was mediated by a hydrolytic process. It began with a lag phase, followed by linear release between 12 and 48 h, and reached equilibrium at 72 h with 51% of the gemcitabine released. In vitro cytotoxicity studies using MCF-7 and MDA-MB-231 human mammary cancer cells, as well as human dermal fibroblasts (HDF), showed that PG-G displayed a lower dose dependent cytotoxic effect with respect to the parent drug gemcitabine. On the other hand, in 4T1 mouse mammary tumour cells, PG-G and gemcitabine showed similar toxicities. Gemcitabine was more than likely released hydrolytically from PG-G and taken up by MCF-7, MDA-MB-231 and HDF, whereas both released gemcitabine and PG-G were taken up by 4T1 to mediate the observed cytotoxicities. The improved stability and extended sustained release profile may render PG-G a potential anticancer prodrug

Rosamines Targeting the Cancer Oxidative Phosphorylation Pathway

Reprogramming of energy metabolism is pivotal to cancer, so mitochondria are potential targets for anticancer therapy. A prior study has demonstrated the anti-proliferative activity of a new class of mitochondria-targeting rosamines. This present study describes in vitro cytotoxicity of second-generation rosamine analogs, their mode of action, and their in vivo efficacies in a tumor allografted mouse model. Here, we showed that these compounds exhibited potent cytotoxicity (average IC50<0.5 µM), inhibited Complex II and ATP synthase activities of the mitochondrial oxidative phosphorylation pathway and induced loss of mitochondrial transmembrane potential. A NCI-60 cell lines screen further indicated that rosamine analogs 4 and 5 exhibited potent antiproliferative effects with Log10GI50 = -7 (GI50 = 0.1 µM) and were more effective against a colorectal cancer sub-panel than other cell lines. Preliminary in vivo studies on 4T1 murine breast cancer-bearing female BALB/c mice indicated that treatment with analog 5 in a single dosing of 5 mg/kg or a schedule dosing of 3 mg/kg once every 2 days for 6 times (q2d×6) exhibited only minimal induction of tumor growth delay. Our results suggest that rosamine analogs may be further developed as mitochondrial targeting agents. Without a doubt proper strategies need to be devised to enhance tumor uptake of rosamines, i.e. by integration to carrier molecules for better therapeutic outcome

The Implicature in Flouting Maxim of Relation by the Main Character in Iron Man 2 Movie

Language takes an important role in human life. It is a tool for communication. People can get what they want and express their feelings by doing communication. In order to have good communication, the speaker and theThis study uses qualitative approach because the writer analyzes conversations in movie. The result of the study shows that there are 7 reasons why the main character flouts the maxim. This study also shows that there are 20 utterances containing flouting maxim of relation. Furthermore, this study alsoreveals that the using of flouting maxim of relation makes conversation moreinteresting.Hopefully, this study will inspire the next researchers to broaden their pointof view in order to look for the problems of the study. Furthermore, the writer alsosuggests the next researchers to look for other subject to be analyzed and use othertheories to analyze the data. Keywords: Cooperative principle, flouting maxim of relation, implicature, iron man 2 movie.hearer have to obey Cooperative Principle. However, there is a phenomenon when occasionally someone says one thing but he/ she means another. It usually happens with flouting maxim. One kind of flouting maxim is flouting maxim of relation. It occurs when a speaker or hearer responses which is very obviously irrelevant to the topic. The phenomenon is presented in the Iron Man 2 movie. This study aims to reveal the reasons why the main character flouts maxim of relation and what the impact to the conversation is

Pegembangan Model untuk Pembelajaran Keterampilan Menulis di Sekolah Dasar

Pembelajaran Bahasa Indonesia seringkali dianggap mudah oleh sebagian besar peserta didik, hal tersebut menjadikan tujuan pembelajaran dalam suatu kegiatan belajar mengajar tidak dapat dicapai secara optimal, salah satunya keterampilan menulis. Banyaknya model pembelajaran yang ditawarkan dalam dunia pendidikan nyatanya belum mampu mengatasi kekurangan tersebut. Tujuan dari penelitian ini adalah 1) untuk mengetahui pembelajaran keterampilan menulis di SD, 2) untuk mengetahui tingkat keterampilan menulis siswa, 3) untuk mengetahui kebutuhan terhadap model pembelajaran Bahasa Indonesia yang dapat meningkatkan keterampilan menulis siswa. Jenis penelitian yang dilakukan adalah penelitian kualitatif yang dilakukan melalui studi kasus dan studi literature. Subjek penelitiannya meliputi siswa kelas III Sekolah Dasar. Teknik pengumpulan data menggunakan triangulasi data/sumber yaitu data wawancara yang dilakukan dengan guru kelas III di sekolah yang berbeda dan triangulasi metode yaitu data wawancara, dan observasi. Analisis data dilakukan mulai dari pengumpulan data, reduksi data, penyajian data sampai verifikasi. Hasil penelitian pada studi pendahuluan ini adalah 1) kurikulum yang diterapkan di empat sekolah tersebut adalah Kurikulum Tingkat Satuan Pendidikan, 2) model pembelajaran yang digunakan guru sudah mengarah pada student center, 3) media yang digunakan berupa gambar yang sudah tertera pada buku, 4) penilaian dilakukan secara menyeluruh (mencakup empat keterampilan), 5)sebagian siswa belum mampu mencapai KKM. Kesimpulan dari studi pendahuluan ini adalah perlu dikembangkannya model pembelajaran Bahasa Indonesia yang dapat meningkatkan keterampilan menulis siswa

Tropomyosin Receptor Kinase C Targeted Delivery of a Peptidomimetic Ligand-Photosensitizer Conjugate Induces Antitumor Immune Responses Following Photodynamic Therapy

Tropomyosin receptor kinase C (TrkC) targeted ligand-photosensitizer construct, IYIY-diiodo-boron-dipyrromethene (IYIY-I(2)-BODIPY) and its scrambled counterpart YIYI-I(2)-BODIPY have been prepared. IYIY-I(2)-BODIPY binds TrkC similar to neurotrophin-3 (NT-3), and NT-3 has been reported to modulate immune responses. Moreover, it could be shown that photodynamic therapy (PDT) elevates antitumor immune responses. This prompted us to investigate the immunological impacts mediated by IYIY-I(2)-BODIPY in pre- and post-PDT conditions. We demonstrated that IYIY-I(2)-BODIPY (strong response) and YIYI-I(2)-BODIPY (weak response) at 10 mg/kg, but not I(2)-BODIPY control, increased the levels of IL-2, IL-4, IL-6 and IL-17, but decreased the levels of systemic immunoregulatory mediators TGF-β, myeloid-derived suppressor cells and regulatory T-cells. Only IYIY-I(2)-BODIPY enhanced the IFN-γ(+) and IL-17(+) T-lymphocytes, and delayed tumor growth (~20% smaller size) in mice when administrated daily for 5 days. All those effects were observed without irradiation; when irradiated (520 nm, 100 J/cm(2), 160 mW/cm(2)) to produce PDT effects (drug-light interval 1 h), IYIY-I(2)-BODIPY induced stronger responses. Moreover, photoirradiated IYIY-I(2)-BODIPY treated mice had high levels of effector T-cells compared to controls. Adoptive transfer of immune cells from IYIY-I(2)-BODIPY-treated survivor mice that were photoirradiated gave significantly delayed tumor growth (~40–50% smaller size) in recipient mice. IYIY-I(2)-BODIPY alone and in combination with PDT modulates the immune response in such a way that tumor growth is suppressed. Unlike immunosuppressive conventional chemotherapy, IYIY-I(2)-BODIPY can act as an immune-stimulatory chemotherapeutic agent with potential applications in clinical cancer treatment

Hemodynamic effects of HPMA copolymer based doxorubicin conjugate: A randomized controlled and comparative spectral study in conscious rats

Conjugation of Doxorubicin (DOX) to N-(2-hydroxypropyl) methylacrylamide copolymer (HPMA) has significantly reduced the DOX-associated cardiotoxicity. However, the reports on the impact of HPMA-DOX conjugates on the cardiovascular system such as blood pressure (BP) and heart rate (HR) were in restrained animals using tail cuff and/or other methods that lacked the resolution and sensitivity. Herein, we employed radiotelemetric-spectral-echocardiography approach to further understand the in vivo cardiovascular hemodynamics and variability post administration of free DOX and HPMA-DOX. Rats implanted with radio-telemetry device were administered intravenously with DOX (5mg/kg), HPMA-DOX (5mg DOX equivalent/kg) and HPMA copolymer and subjected to continuous cardiovascular monitoring and echocardiography for 140 days. We found that DOX-treated rats had ruffled fur, reduced body weight (BW) and a low survival rate. Although BP and HR were normal, spectral analysis indicated that their BP and HR variabilities were reduced. All rats exhibited typical signs of cardiotoxicity at histopathology. In contrast, HPMA-DOX rats gained weight over time and survived. Although BP, HR and related variabilities were unaffected, the left ventricular end diastolic volume (EDV) of these rats, as well as of the HPMA copolymer-treated rats, was found increased at the end of observation period. Additionally, HPMA copolymer caused microscopic injury of the heart tissue. All of these suggest the necessity of caution when employing HPMA as carrier for prolonged drug delivery. The current study also indicates the potential of radiotelemetric-spectral-echocardiography approach for improved preclinical cardiovascular risk assessment of polymer-drug conjugate and other nano-sized-drug constructs

Hemodynamic effects of HPMA copolymer based doxorubicin conjugate:A randomized controlled and comparative spectral study in conscious rats

<p>Conjugation of Doxorubicin (DOX) to <i>N</i>-(2-hydroxypropyl) methylacrylamide copolymer (HPMA) has significantly reduced the DOX-associated cardiotoxicity. However, the reports on the impact of HPMA–DOX conjugates on the cardiovascular system such as blood pressure (BP) and heart rate (HR) were in restrained animals using tail cuff and/or other methods that lacked the resolution and sensitivity. Herein, we employed radiotelemetric-spectral-echocardiography approach to further understand the <i>in vivo</i> cardiovascular hemodynamics and variability post administration of free DOX and HPMA–DOX. Rats implanted with radio-telemetry device were administered intravenously with DOX (5 mg/kg), HPMA–DOX (5 mg DOX equivalent/kg) and HPMA copolymer and subjected to continuous cardiovascular monitoring and echocardiography for 140 days. We found that DOX-treated rats had ruffled fur, reduced body weight (BW) and a low survival rate. Although BP and HR were normal, spectral analysis indicated that their BP and HR variabilities were reduced. All rats exhibited typical signs of cardiotoxicity at histopathology. In contrast, HPMA–DOX rats gained weight over time and survived. Although BP, HR and related variabilities were unaffected, the left ventricular end diastolic volume (EDV) of these rats, as well as of the HPMA copolymer-treated rats, was found increased at the end of observation period. Additionally, HPMA copolymer caused microscopic injury of the heart tissue. All of these suggest the necessity of caution when employing HPMA as carrier for prolonged drug delivery. The current study also indicates the potential of radiotelemetric-spectral-echocardiography approach for improved preclinical cardiovascular risk assessment of polymer–drug conjugate and other nano-sized-drug constructs.</p