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Citation: Takayama K, Kaneko H, Hwang S-J, et al. Increased ocular levels of microRNA-148a in cases of retinal detachment promote epithelialmesenchymal transition. Invest Ophthalmol Vis Sci. 2016;57:269957: -270557: . DOI:10.1167 PURPOSE. The purpose of this study was to determine microRNA expression in vitreous and subretinal fluid (SRF) samples from patients with retinal detachment (RD). The pathological importance of the identified microRNA transcript levels was analyzed in vitro.METHODS. Vitreous fluid was collected from 10 patients with macular hole (MH), vitreomacular traction syndrome (VMTS), or foveoschisis and from 11 patients with RD. Subretinal fluid was collected from 7 patients with RD. Of these, blood serum was collected in 4 patients. MicroRNA microarray profiling was performed to identify microRNA transcripts that were present in vitreous fluid, and more redundantly detected in SRF, of patients with RD, but not detected in control eyes. Western blotting and scratch assays were performed in ARPE-19 cells and primary human RPE cell lines transfected with microRNA to elucidate the effect of identified microRNA transcripts on epithelial-mesenchymal transition (EMT). RESULTS.MicroRNA microarray profiling revealed that hsa-miR-148a-3p was the most redundantly detected transcript in SRF and vitreous fluid from patients with RD, but not those with the other diseases. Expression levels of hsa-miR-148a-3p were higher in SRF samples than in blood serum samples in 3 out of 4 patients. Following hsa-miR-148a-3p mimic transfection, ARPE-19 and human RPE cells demonstrated increased expression of a-smooth muscle actin by Western blotting and increased migration ability during scratch assays. CONCLUSIONS.The results of the present study indicate that hsa-miR-148a-3p was specifically detected in RD and promotes EMT in RPE.

Contrast-enhanced ultrasonography has an important role in the detection of tumors in humans. The second-generation contrast agent Sonazoid has the ability of real-time contrast imaging along with parenchymal imaging. The purposes of this study were to determine the effect and duration of Sonazoid on the changes in gray-scale enhancement of canine spleen and to establish an appropriate protocol for contrast-enhanced ultrasonography of canine spleen. Six healthy beagles were injected with an intravenous bolus of Sonazoid. In the spleen parenchyma, the enhancement was maintained up to 30 min after injection. Moreover, for 5-22 s after injection, gray-scale enhancement of splenic arteries afforded arterial imaging. Perfusion of the kidney may be investigated from 3.6s to 3.5 min after injection of Sonazoid. These results suggest that Sonazoid is applicable to canine spleen parenchymal imaging and that the optimal time for the parenchymal imaging is 7-30 min after injection. The findings of this quantitative study should prove useful in the evaluation of diffuse or focal splenic and renal diseases in dogs.

Silicone oil (SO) is an intraocular surgical adjuvant that reduces the surgical complications in refractory retinal diseases, although membrane and cellular proliferation is often seen even in SO-filled eyes. We hypothesised that the fluid in the space between the SO and the retina, named the “sub-silicone oil fluid (SOF)”, enhances these biological responses. We proposed a safe method for SOF extraction. We also analysed inflammatory cytokine expressions and SOF osmotic pressures from eyes with rhegmatogenous retinal detachment (RRD), proliferative diabetic retinopathy (PDR), proliferative vitreoretinopathy (PVR) and macular hole-associated retinal detachment (MHRD). Interleukin (IL)-10, IL-12p40, IL-6, monocyte chemotactic protein-1, and vascular endothelial growth factor (VEGF) in the SOF with PVR were significantly higher than in those with RRD or MHRD. Fibroblast growth factor-2, IL-10, IL-12p40, IL-8, VEGF, and transforming growth factor beta 1 levels in eyes with exacerbated PDR indicated a significantly higher expression than those with simple PDR. IL-6 and tumour necrosis factor alpha in eyes with exacerbated PVR demonstrated a significantly higher expression than in those with simple PVR. However, there was no difference in SOF osmotic pressure between group of each disease. These studies indicate that disease-specific SOF is a significant reflection of disease status.

ABSTRACT. We attempted to develop a strain of Babesia gibsoni resistant to diminazene aceturate (DA), an anti-babesial drug, in vitro. Since the DA-sensitive B. gibsoni strain could survive and proliferate in culture medium containing 1 ng/ml DA, the concentration of DA was gradually increased from 1 to 200 ng/ml. The results showed that the parasites could survive and proliferate in the medium containing 200 ng/ml DA, which was much higher than the 50% inhibitory concentration (IC 50 ) of DA for B. gibsoni. Subsequently, these parasites were removed from erythrocytes and exposed directly to 200 ng/ml DA. They were able to survive and invade fresh erythrocytes, though the DA-sensitive B. gibsoni strain did not survive. Based on these results, the parasites cultured within 200 ng/ml DA were determined to be a DA-resistant B. gibsoni strain. In addition, the IC 50 levels of clindamycin, doxycycline and pentamidine for the DA-resistant B. gibsoni strain were determined. The IC 50 levels of clindamycin, doxycycline and pentamidine for the DA-resistant strain were higher than those for the DA-sensitive strain. The IC 50 of pentamidine for the resistant strain was much greater than that for the DA-sensitive strain. These results indicated that the DA-resistant B. gibsoni strain could have resistance not only to DA, but also to other anti-babesial drugs. In conclusion, we successfully developed a DA-resistant B. gibsoni strain in vitro. Canine babesiosis, a tick-borne hematozoan disease, is caused by Babesia gibsoni and B. canis. This disease is characterized by fever, lethargy, anemia and, in severe cases, death [9,11]. Diminazene aceturate (DA), an antibabesial drug, is an aromatic diamidine derivative. Currently, the mechanism of action of DA on B. gibsoni and B. canis is unknown. DA can temporarily improve the clinical signs of canine babesiosis [4,13]. However, this drug is unable to eliminate the parasites from infected dogs, and relapses often occur [11]. We believe that this is due to the development of drug resistance of B. gibsoni against DA. Collett [8] considered that B. canis surviving DA treatment could develop drug resistance against DA clinically. However, there is no report proving DA resistance of B. gibsoni and B. canis. In trypanosomiasis and leishmaniasis, it has been reported that DA can inhibit the DNA replication and mitochondrial respiratory activity of these pathogens [3,16]. The loss of P2 nucleoside transporter function in Trypanosoma brucei brucei has been implicated in resistance to DA [6]. Likewise, it is possible that B. gibsoni could develop drug resistance against DA.In reports about other anti-babesial drugs, including atovaquone, clindamycin, metronidazole, doxycycline and pentamidine, almost no single drug treatment or combined treatment could eliminate the parasites from the peripheral blood at the dosages used, and the possibility of relapse and development of resistant variants remained [11,13,24,27,32]. From those previous reports, it seems to be difficult to eliminate B. gibson...