Isolation of rat hepatocytes for pharmacological studies on metabotropic glutamate receptor (mGluR) subtype 5: a comparison between collagenase I versus collagenase IV.

Isolated hepatocytes can be obtained from the liver by collagenase infusion, a procedure that could affect cell isolation as well as the integrity of membrane receptors. In this respect we compared metabotropic glutamate subtype 5 receptor (mGluR5) protein expression and activity in rat hepatocytes isolated by two collagenases, type I and type IV. Hepatocytes were isolated from male Wistar rats (200-250 g) using collagenase I or colla-genase IV and after isolation, viability and morphology of rat hepatocytes were assessed measuring mGluR5 protein expression by Western blot analyses. mGluR5 activation was evaluated by inositol-1-phosphate (IP-1) accumulation after treatment with the mGluR5 orthosteric agonist ACPD or the selective antagonist MPEP. No difference in cellular viability and morphology was observed using collagenase I when compared with collage-nase IV. An increase in mGluR5 protein expression was observed in hepatocytes isolated using collagenase IV with respect to collagenase I. Moreover, hepatocytes treated with ACPD and with MPEP presented higher levels of IP-1 when isolated using collagenase IV compared tocollagenase I. These results indicate that collage-nase IV better preserves the activity of surface proteins such as mGluR5in isolated rat hepatocytes, an in vitro model useful to reduce the use of experimental animals, in line with the 3R ethical framework

Selective blockade of mglu5 metabotropic glutamate receptors is protective against acetaminophen hepatotoxicity in mice

BACKGROUND/AIMS: mGlu5 metabotropic glutamate receptor antagonists protect rat hepatocytes against hypoxic death. Here, we have examined whether mGlu5 receptor antagonists are protective against liver damage induced by oxidative stress. METHODS: Toxicity of isolated hepatocytes was induced by tert-butylhydroperoxide (t-BuOOH) after pretreatment with the mGlu5 receptor antagonists, MPEP, SIB-1757 and SIB-1893. The effect of these drugs was also examined in mice challenged with toxic doses of acetaminophen. RESULTS: Addition of tBuOOH (0.5 mM) to isolated hepatocytes induced cell death (70+/-5% at 3 h). Addition of MPEP or SIB-1893 to hepatocytes reduced both the production of reactive oxygen species (ROS) and cell toxicity induced by t-BuOOH (tBuOOH=70+/-5%; tBuOOH+MPEP=57+/-6%; tBuOOH+SIB-1893=40+/-4%). In mice, a single injection of acetaminophen (300 mg/kg, i.p.) induced centrilobular liver necrosis, which was detectable after 24 h. MPEP (20 mg/kg, i.p.) substantially reduced liver necrosis and the production of ROS, although it did not affect the conversion of acetaminophen into the toxic metabolite, N-acetylbenzoquinoneimine. MPEP, SIB-1893 and SIB-1757 (all at 20 mg/kg, i.p.) also reduced the increased expression and activity of liver iNOS induced by acetaminophen. CONCLUSIONS: We conclude that pharmacological blockade of mGlu5 receptors might represent a novel target for the treatment of drug-induced liver damage

Transient expression of reck under hepatic ischemia/reperfusion conditions is associated with mapk signaling pathways

In this study, we demonstrated the involvement of matrix metalloproteinases (MMPs) in hepatic ischemia/reperfusion (I/R) injury. Our aim is to evaluate the impact of reperfusion on I/R-related changes in RECK, an MMP modulator, and mitogen-activated protein kinase (MAPKs) pathways (ERK, p38, and JNK). Male Wistar rats were either subjected to 60 min partial-hepatic ischemia or sham-operated. After a 60 min or 120 min reperfusion, liver samples were collected for analysis of MMP-2 and MMP-9 by zymography and RECK, TIMP-1, and TIMP-2 content, MAPKs activation (ERK1/2, JNK1/2, and p38), as well as iNOS and eNOS by Western blot. Serum enzymes AST, ALT, and alkaline-phosphatase were quantified. A transitory decrease in hepatic RECK and TIMPs was associated with a transitory increase in both MMP-2 and MMP-9 activity and a robust activation of ERK1/2, JNK1/2, and p38 were detected at 60 min reperfusion. Hepatic expression of iNOS was maximally upregulated at 120 min reperfusion. An increase in eNOS was detected at 120 min reperfusion. I/R evoked significant hepatic injury in a time-dependent manner. These findings provide new insights into the underlying molecular mechanisms of reperfusion in inducing hepatic injury: a transitory decrease in RECK and TIMPs and increases in both MAPK and MMP activity suggest their role as triggering factors of the organ dysfunction

Assessment of peloids used for the Spa therapy.

Nella Medicina Termale i fanghi o peloidi sono sicuramente tra i presidi terapeutici più utilizzati. Gli effetti della loro applicazione sono suddivisi in locali (nella sede d'applicazione) e generali (a livello dell' intero organismo). Gli effetti locali originano da stimoli indotti dalla fangoterapia su singoli distretti corporei, ma determinano una risposta di tutto l'organismo ed il coinvolgimento di numerosi organi, apparati e sistemi metabolici. A loro volta le reazioni generali indotte dalla fango terapia possono influire sulla risposta locale. Vi è quindi un rapporto biunivoco tra i due meccanismi ed una serie di azioni e reazioni. Un' altra possibile suddivisione è quella che distingue gli effetti specifici e aspecifici della fangoterapia. I primi sono legati per lo più al particolare tipo di acqua minerale contenuta nel fango (azione attribuita ai singoli mineralizzatori ed ai gas disciolti nell' acqua) e alla $ microflora che in esso si sviluppa durante la maturazione. I secondi effetti, aspecifici, sono da ricondursi sia all'azione biologica e terapeutica del calore (termoterapia) che alle altre proprietà fisiche proprie della metodica e del mezzo impiegato. Questi effetti specifici ed aspecifici possono a loro volta esplicarsi a livello locale e generale. Il risultato terapeutico complessivo della fangoterapia è comunque influenzato in parte dalla reattività individuale del paziente ed in parte dipende dal tipo di fango e dalle modalità di applicazione. Gli effetti biologici e terapeutici sono fondamentalmente riconducibili a: - azione antinfiammatoria; azione analgesica; - azione miorilassante; - azione eutrofica; - aumento delle resistenze verso le noxae patogene esogene ed endogene; - azione di stimolo in numerosi processi metabolici

Recent biotechnological developments in the use of peroxidases

Peroxidases are ubiquitous oxidoreductases that use hydrogen peroxide or alkyl peroxides as oxidants. Advances have recently been made in using them to prepare, under mild and controlled conditions, chiral organic molecules that are valuable for the chemoenzymatic synthesis of a wide range of useful compounds. Horseradish peroxidase can be converted into a peroxygenative enzyme by molecular engineering. Chloroperoxidase, the most versatile peroxidase, behaves like a 'true' monooxygenase in sulfoxidations with molecular oxygen and an external reductant, with substantial increases in enantioselectivity and enzyme stability

Effects of a salsobromoiodine water on patients treated with photodynamic therapy for acne

Skripsi ini mengkaji tentang Arsitektur Masjid Jami’ Gresik. Adapun masalah yang dibahas pada skripsi ini adalah sebagai berikut: 1) Bagaimana latar belakang fungsi pembangunan Masjid Jami’? 2) Bagaimana dinamika peradaban Islam diwilayah sekitar masjid Jami’ Gresik? 3) Apa makna yang terdapat dalam arsitektural Masjid Jami’ Gresik ? Untuk menjawab permasalahan tersebut, penulis menggunakan metode sejarah. Metode ini menggunakan empat tahap penelitian yaitu, Heuristik (Pengumpulan sumber), Verifikasi (Kritik Sumber), Interpretasi (Penafsiran Sumber), dan Historiografi (Penulisan Sejarah). Penelitian ini menggunakan pendekatan etnografi, pendekatan ini bertujuan mendapatkan deskripsi dan analisis mendalam berkaitan dengan perubahan sosial dan kebudayaan manusia. Serta menyajikan fakta secara sistematis tentang obyek yang diteliti yaitu Analisis, Makna dan Simbol dari Arsitektur Masjid Jami’ Gresik. Adapun teori yang digunakan dalam skripsi ini adalah Continuity and Change. Dengan teori ini penulis berharap dapat menguraikan analisis, makna dan simbol arsitektur Masjid Jami’ Gresik. Dengan rumusan yang ada serta dari beberapa penelusuran yang yang penuis lakukan dari sumber-sumber primer dan sekunder, membuktikan bahwa: 1) Masjid Jami’Gresik awalnya merupakan mushola kecil yang dibangun pada masa islamisasi Maulana Malik Ibrahim. Awal mulanya, latar belakang fungsi pembangunan Masjid Jami’Gresik adalah tidak lain untuk menampung jamaah yang semakin bertambah banyak dan tersebar diberbagai wilayah; 2) Masjid Jami’ Gresik merupakan bagian dari tata letak kota yang sudah ada sejak masa awal penyebaran Islam di Indonesia; 3) Masjid Jami’ Gresik merupakan salah satu masjid tua yang ada di Kabupaten Gresik, keberadaannya dijadikan sebagai warisan budaya