AAV2-Mediated Subretinal Gene Transfer of hIFN-α Attenuates Experimental Autoimmune Uveoretinitis in Mice

BACKGROUND: Recent reports show that gene therapy may provide a long-term, safe and effective intervention for human diseases. In this study, we investigated the effectiveness of adeno-associated virus 2 (AAV2) based human interferon-alpha (hIFN-α) gene therapy in experimental autoimmune uveoretinitis (EAU), a classic model for human uveitis. METHODOLOGY/PRINCIPAL FINDINGS: An AAV2 vector harboring the hIFN-α gene (AAV2.hIFN-α) was subretinally injected into B10RIII mice at two doses (1.5×10(6) vg, 1.5×10(8) vg). AAV2 vector encoding green fluorescent protein (AAV2.GFP) was used as a control (5×10(8) vg). The expression of hIFN-α in homogenized eyes and serum was detected by ELISA three weeks after injection. The biodistribution of vector DNA in the injected eyes, contralateral eyes and distant organs was determined by PCR. EAU was induced by immunization with IRBP(161-180) three weeks following vector injections, and evaluated clinically and pathologically. IRBP-specific proliferation and IL-17 expression of lymphocytes from the spleen and lymph nodes were assayed to test the influence of the subretinal delivery of AAV2.hIFN-α on the systemic immune response. hIFN-α was effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2.hIFN-α vector. DNA of AAV2.GFP was observed only in the injected eyes, but not in the distant organs or contralateral eyes. Subretinal injection of both doses significantly attenuated EAU activity clinically and histologically. For the lower dose, there was no difference concerning lymphocyte proliferation and IL-17 production among the AAV2.hIFN-α, AAV2.GFP and PBS injected mice. However, the higher dose of AAV2.hIFN-α significantly suppressed lymphocyte proliferation and IL-17 production. CONCLUSIONS/SIGNIFICANCE: Subretinal delivery of AAV2.hIFN-α lead to an effective expression within the eye for at least three months and significantly attenuated EAU activity. AAV2.hIFN-α was shown to inhibit the systemic IRBP-specific immune response

Head Exposure to Cold during Whole-Body Cryostimulation: Influence on Thermal Response and Autonomic Modulation

Recent research on whole-body cryotherapy has hypothesized a major responsibility of head cooling in the physiological changes classically reported after a cryostimulation session. The aim of this experiment was to verify this hypothesis by studying the influence of exposing the head to cold during whole-body cryostimulation sessions, on the thermal response and the autonomic nervous system (ANS). Over five consecutive days, two groups of 10 participants performed one whole-body cryostimulation session daily, in one of two different systems; one exposing the whole-body to cold (whole-body cryostimulation, WBC), and the other exposing the whole-body except the head (partial-body cryostimulation, PBC).10 participants constituted a control group (CON) not receiving any cryostimulation. In order to isolate the head-cooling effect on recorded variables, it was ensured that the WBC and PBC systems induced the same decrease in skin temperature for all body regions (mean decrease over the 5 exposures: -8.6°C±1.3°C and -8.3±0.7°C for WBC and PBC, respectively), which persisted up to 20-min after the sessions (P20). The WBC sessions caused an almost certain decrease in tympanic temperature from Pre to P20 (-0.28 ±0.11°C), while it only decreased at P20 (-0.14±0.05°C) after PBC sessions. Heart rate almost certainly decreased after PBC (-8.6%) and WBC (-12.3%) sessions. Resting vagal-related heart rate variability indices (the root-mean square difference of successive normal R-R intervals, RMSSD, and high frequency band, HF) were very likely to almost certainly increased after PBC (RMSSD:+49.1%, HF: +123.3%) and WBC (RMSSD: +38.8%, HF:+70.3%). Plasma norepinephrine concentration was likely increased in similar proportions after PBC and WBC, but only after the first session. Both cryostimulation techniques stimulated the ANS with a predominance of parasympathetic tone activation from the first to the fifth session and in slightly greater proportion with WBC than PBC. The main result of this study indicates that the head exposure to cold during whole-body cryostimulation may not be the main factor responsible for the effects of cryostimulation on the ANS

GFAP-Driven GFP Expression in Activated Mouse Muller Glial Cells Aligning Retinal Blood Vessels Following Intravitreal Injection of AAV2/6 Vectors

Background: Muller cell gliosis occurs in various retinal pathologies regardless of the underlying cellular defect. Because activated Muller glial cells span the entire retina and align areas of injury, they are ideal targets for therapeutic strategies, including gene therapy.Methodology/Principal Findings: We used adeno-associated viral AAV2/6 vectors to transduce mouse retinas. The transduction pattern of AAV2/6 was investigated by studying expression of the green fluorescent protein (GFP) transgene using scanning-laser ophthalmoscopy and immuno-histochemistry. AAV2/6 vectors transduced mouse Muller glial cells aligning the retinal blood vessels. However, the transduction capacity was hindered by the inner limiting membrane (ILM) and besides Muller glial cells, several other inner retinal cell types were transduced. To obtain Muller glial cell-specific transgene expression, the cytomegalovirus (CMV) promoter was replaced by the glial fibrillary acidic protein (GFAP) promoter. Specificity and activation of the GFAP promoter was tested in a mouse model for retinal gliosis. Mice deficient for Crumbs homologue 1 (CRB1) develop gliosis after light exposure. Light exposure of Crb1(-/-) retinas transduced with AAV2/6-GFAP-GFP induced GFP expression restricted to activated Muller glial cells aligning retinal blood vessels.Conclusions/Significance: Our experiments indicate that AAV2 vectors carrying the GFAP promoter are a promising tool for specific expression of transgenes in activated glial cells

Parasympathetic Activity and Blood Catecholamine Responses Following a Single Partial-Body Cryostimulation and a Whole-Body Cryostimulation

The aim of this study was to compare the effects of a single whole-body cryostimulation (WBC) and a partial-body cryostimulation (PBC) (i.e., not exposing the head to cold) on indices of parasympathetic activity and blood catecholamines. Two groups of 15 participants were assigned either to a 3-min WBC or PBC session, while 10 participants constituted a control group (CON) not receiving any cryostimulation. Changes in thermal, physiological and subjective variables were recorded before and during the 20-min after each cryostimulation. According to a qualitative statistical analysis, an almost certain decrease in skin temperature was reported for all body regions immediately after the WBC (mean decrease±90% CL, -13.7±0.7°C) and PBC (-8.3±0.3°C), which persisted up to 20-min after the session. The tympanic temperature almost certainly decreased only after the WBC session (-0.32±0.04°C). Systolic and diastolic blood pressures were very likely increased after the WBC session, whereas these changes were trivial in the other groups. In addition, heart rate almost certainly decreased after PBC (-10.9%) and WBC (-15.2%) sessions, in a likely greater proportion for WBC compared to PBC. Resting vagal-related heart rate variability indices (the root-mean square difference of successive normal R-R intervals, RMSSD, and high frequency band, HF) were very likely increased after PBC (RMSSD: +54.4%, HF: +138%) and WBC (RMSSD: +85.2%, HF: +632%) sessions without any marked difference between groups. Plasma norepinephrine concentrations were likely to very likely increased after PBC (+57.4%) and WBC (+76.2%), respectively. Finally, cold and comfort sensations were almost certainly altered after WBC and PBC, sensation of discomfort being likely more pronounced after WBC than PBC. Both acute cryostimulation techniques effectively stimulated the autonomic nervous system (ANS), with a predominance of parasympathetic tone activation. The results of this study also suggest that a whole-body cold exposure induced a larger stimulation of the ANS compared to partial-body cold exposure

EFEKTIVITAS PENGAWASAN UNIT KERJA ANTI FRAUD PADA BANK MUAMALAT INDONESIA

Perkembangan perbankan syari‟ah di Indonesia demikian pesat yang ditandai dengan berdirinya Bank Muamalat Indonesia. Perkembangan ini berimplikasi pada besarnya tantangan perbankan syari‟ah di Indonesia terutama dalam mempertahankan identitasnya sebagai perusahaan yang bergerak berlandaskan prinsip-prinsip syari‟ah. Sejak berdirinya perbankan syariah,berbagai kontroversi muncul dari masyarakat, masalah yang paling banyak disorot adalah pelekatan label syariah pada institusi keuangan Islam yang masih dianggap belum layak. Keraguan masyarakat tersebut seolah terjawab dengan munculnya kasus yang cukup menggemparkan yakni kasus fraud (tindak kecurangan) yang terjadi di lembaga syariah. Bank Muamalat Indonesia merupakan bank syari‟ah pertama yang muncul dengan gagasan bank murni syari‟ah. Akan tetapi, bank Muamalat Indonesia juga tak luput dari kasus fraud yang dilakukan oleh karyawan bank tersebut. Berdasarkan Laporan Tahunan BMI menyebutkan bahwa telah terjadi peningkatan kasus fraud dari tahun sebelumnya yang berjumlah 18 kasus menjadi 82 kasus pada tahun 2016. Padahal perusahaan yang menggunakan identitas syariah seharusnya dapat lebih meminimalisir bahkan meniadakan resiko terjadinya kasus fraud dengan adanya internal control perusahaan. Dari latar belakang tersebut, peneliti berusaha mendalami peran pengawasan Unit Kerja Anti Fraud dalam fraud preventive pada Bank Muamalat Indonesia. Penelitian ini merupakan penelitin pustaka yang bersifat deskriptif analisis dengan pendekatan kualitatif. Adapun sumber bahan hukum primer yang dipakai yaitu berdasarkan Laporan Tahunan Bank Muamalat Indonesia Tahun 2016. Sedangkan sumber bahan hukum sekunder berupa buku-buku, jurnal,karya ilmiah, artikel, terkait dengan strategi anti fraud perbankan syariah. Dari hasil penelitian dikemukakan bahwa peningkatan kasus fraud yang terjadi pada Bank Muamalat Indonesia disebabkan kurang efektifnya pengawasan Unit Kerja Anti Fraud. Hal ini dikarenakan kegiatan yang dilakukan selama tahun 2016 belum menujukkan adanya usaha preventif terhadap kasus fraud. Sedangkan pencegahan merupakan pilar penting dalam keefektivan sebuah pengawasan. Tujuan perusahaan dalam mencegah fraud dapat tercapai, jika fungsi pengawasan dilakukan sebelum terjadinya penyimpangan-penyimpangan sehingga lebih bersifat mencegah (prefentive control). Oleh karena itu, keefektivan pengawasan Unit Kerja Anti Fraud diharapkan dapat meminimalisir tindak kecurangan demi mewujudkan perusahaan yang patuh terhadap ketentuan syariah sesuai dengan identitas perusahaan. vii Usaha pencegahan terjadinya kasus pada Bank Muamalat Indonesia diharapkan dapat menjadi bukti terlaksananya tatakelola perusahaan (Good Corporate Governance) pada Bank Syari‟ah dengan baik. Hal ini berdasarkan dalam dalam perbankan syariah dikenal adanya prinsip-prinsip syariah yang mendukung bagi terlaksananya prinsip GCG yakni keharusan bagi subjek hukum termasuk bank untuk menerapkan prinsip kejujuran (shiddiq), edukasi kepada masyarakat (tabligh), kepercayaan (amanah), dan pengelolaan secara profesional (fathanah)

2017 HRS/EHRA/ECAS/APHRS/SOLAECE expert consensus statement on catheter and surgical ablation of atrial fibrillation: executive summary.

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