Seminal Vesicle Involvement by Carcinoma In Situ of the Bladder: Clonal Analysis Using Next-Generation Sequencing to Elucidate the Mechanism of Tumor Spread / Journal of the Korean Cancer Association, 대한암학회지

We present a rare case of urothelial carcinoma in situ (CIS), which invades the prostate and seminal vesicle (SV). A 70-year-old man underwent transurethral resection of bladder (TURB), and the pathologic examination revealed multiple CIS. Although the patient received intravesical bacillus Calmette-Guerin (BCG) therapy following TURB, recurrence of CIS was confirmed in the bladder and left distal ureter at 3 months following BCG. Radical cystectomy was performed due to BCG-refractory CIS. Microscopically, CIS was found throughout the mucosa of the bladder, left ureter, prostatic duct, and both SVs. Next-generation sequencing revealed significant differences in tumor clonality between bladder and SV CIS cells. Among 101 (bladder CIS) and 95 (SV CIS) somatic mutations, only two were shared, and only one gene (ARHGAP23) was common exon coding region gene. In conclusion, multicentric genetic changes, in line with the field-cancerization effect, may result in SV involvement by CIS of the bladder.

Analysis of Changes in Retinal Photoreceptors Using Optical Coherence Tomography in a Feline Model of Iodoacetic Acid-induced Retinal Degeneration

PURPOSE: We investigated structural changes in the retina by using optical coherence tomography (OCT) in a feline model of retinal degeneration using iodoacetic acid (IAA).METHODS: We examined 22 eyes of 11 felines over 2 years of age. The felines had fasted for 12 hours and were intravenously injected with IAA 20 mg/kg of body weight. OCT (Spectralis OCT) was performed at the point where the ends of the retinal vessels collected in the lateral direction from the optic nerve head and area centralis. Similarly, OCT was performed four times at 1-week intervals following injections, at which point the felines were sacrificed and histologic examinations were performed. Using OCT, the thickness of each layer of the retina was measured.RESULTS: The average body weight of the three male and eight female felines investigated in this study was 1.61 ± 0.19 kg. The mean total retinal thickness of the felines before injection was 221.32 ± 9.82 µm, with a significant decrease in the retinal thickness at 2, 3, and 4 weeks following injections of 186.41 ± 35.42, 174.56 ± 31.94, and 175.35 ± 33.84 µm, respectively (p = 0.028, 0.027, and 0.027, respectively). The thickness of the outer nuclear layer was 57.49 ± 8.03 µm before injection and 29.26 ± 17.87, 25.62 ± 13.88, and 31.60 ± 18.38 µm at 2, 3, and 4 weeks, respectively, after injection (p = 0.028, 0.028, 0.046, respectively).CONCLUSIONS: In a feline model of retinal degeneration using IAA, the total retinal thickness and the thickness of the outer nuclear layer were shown to decrease significantly on OCT.

Rapid Progression of the Epiretinal Membrane after Intravitreal Aflibercept Injection

PURPOSE: To report a case of rapid progression of the epiretinal membrane following intravitreal aflibercept injection in a patient with exudative age-related macular degeneration. CASE SUMMARY: An 82-year-old female presented with a complaint of decreased visual acuity in her left eye for 1 month. The initial best-corrected visual acuity was 0.2 in the left eye. Fundus examination and optical coherence tomography revealed soft drusen with retinal hemorrhage of the macula and a transparent epiretinal membrane in the left eye. Fluorescein angiography and indocyanine green angiography showed retinal angiomatous proliferation (RAP) of the left eye, so intravitreal aflibercept injection was performed. One month after the first injection, intraretinal cystic macular edema decreased, while transparency of the epiretinal membrane decreased and reflectivity and thickness of the membrane increased. After two additional injections of aflibercept, RAP showed improvement, whereas the epiretinal membrane progressed. Visual acuity of the left eye decreased to 0.1 and vitrectomy of the membrane was performed. CONCLUSIONS: Careful observation for potentially rapid progression of epiretinal membrane is needed after intravitreal aflibercept injection in the management of exudate age-related macular degeneration accompanied by epiretinal membrane.

National Practice Pattern and Time Trends in Treatment of Upper Urinary Tract Calculi in Korea: a Nationwide Population-Based Study

Despite high prevalence of upper urinary tract calculi (UUTC), there are few studies regarding patterns of care in Asian populations. We investigated treatment patterns and time trends in patients with newly diagnosed UUTC in Korea using the National Health Insurance database that includes de-identified claims from a random 2% sample of the entire population (> 1 million people). A total of 14,282 patients who received active treatments, including shock wave lithotripsy (SWL), ureteroscopic surgery (URS), percutaneous nephrolithotomy (PNL), and uretero/pyelolithotomy (UPL), for newly diagnosed UUTC between 2003 and 2013 were included. The number of primary and all treated cases of UUTC significantly (43% and 103.3%, respectively) increased over the 10-year period. While patients undergoing SWL, URS, PNL, and UPL as primary treatment increased by 43.7%, 31.9%, 87.5%, and 0%, respectively, the relative proportion undergoing each treatment remained constant over the 10 years (SWL > 90%, URS 4.5% to 7.8%, PNL 0.4% to 1.0%, and UPL 40 years (compared to age < 30 years) was significantly associated with URS, PNL, and UPL, rather than SWL, while patients living in urban or suburban/rural areas (compared to metropolitan) were significantly less likely to undergo URS and PNL. In summary, the majority of Korean patients underwent SWL as primary treatment for UUTC, and the predominant use of SWL remained steady over a 10-year period in Korea. Our results will be valuable in examining treatment patterns and time trends in Korean UUTC patients.

Clinical Characteristics and Prognostic Factors in Pediatric Chronic Rhinosinusitis with Polyps / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Chronic rhinosinusitis with nasal polyps is an uncommon pathology in the pediatric population and a challenging problem to otolaryngologists. In this study, we aimed to assess the clinical characteristics and postoperative results of children who underwent sinus surgery due to nasal polyps. SUBJECTS AND METHOD: We retrospectively reviewed medical records of 45 pediatric patients who had sinus surgery from 2009 to 2012. We studied the relationship between clinical parameters and postoperative results. RESULTS: Forty-five patients (18 women and 27 men with an age range of 8 to 17 years) were treated surgically in our hospital. We found statistically significant correlation between the preoperative CT scores (p=0.043), the nasal obstruction symptom scores (p=0.032) and postoperative recurrence, but not between other parameters. CONCLUSION: In this study, prognostic factors affecting the postoperative outcome were preoperative CT score and nasal obstruction symptom score.

Analysis of the Expression Pattern of microRNA of KSHV in KSHV-infected Human Cells

Kaposi's sarcoma associated herpesvirus (KSHV) is subdivided into gamma-herpesvirus and causes Kaposi's sarcoma in human immunodeficiency virus (HIV)-infected patients. A defining feature of herpesviral biology is the presence of two alternative genetic lifestyles - a latent infection and a lytic replicative cycle. Almost all herpesviruses examined so far have been shown to express viral miRNAs in latently and/or productively infected cells. KSHV encodes an array of 15 distinct miRNAs, all of which are expressed at readily detectable levels in latently KSHV infected cells. The expression of an array of these viral miRNAs in KSHV-infected cells suggests that down-regulation of host cell mRNAs by miRNA-mediated RNA interference may represent a critical step in the establishment and/or maintenance of latent infections by KSHV. To investigate KSHV miRNAs that are expressed in KSHV-infected cells, KSHV-infected human umbilical cord vein endothelial cells (HUVECs) and BCBL-1 cells were used and their miRNAs were analyzed by a modified real-time PCR method. Some KSHV miRNAs were detected in KSHV-infected HUVECs and their expression was affected by genetic life cycles. In addition, KSHV miRNAs were also detected in BCBL-1 and their expression was not related to treatment of sodium butyrate. These results indicate that KSHV infection in cells inducing KSHV miRNAs expression would be increased upon entry into latent replication.

Novel Swine-Origin Influenza A (H1N1) Viral Encephalitis

The World Health Organization declared that a new strain of novel swine-origin influenza A (H1N1) virus was responsible for the pandemic infection in June 2009. We report a case of encephalitis diagnosed as the H1N1 virus infection. We describe a 17-year-old patient who had a seizure attack, diagnosed with a H1N1 virus infection via real time reverse-transcriptase polymerase chain reaction (RT-PCR). The H1N1 virus infection can be causative of the encephalitis, as with other influenza virus infections. Careful monitoring is essential for reducing complications.

Establishement of Antibody Selection by Ribosome Display

BACKGROUND: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than 109 members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to 1014 molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. METHODS: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. RESULTS: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. CONCLUSION: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

Terminal Protein-specific scFv Production by Phage Display

BACKGROUND: One of the important factors in the prognosis of chronic hepatitis B patient is the degree of replication of hepatitis B virus (HBV). It has been known that HBV DNA polymerase plays the essential role in the replication of HBV. HBV DNA polymerase is composed of four domains, TP (Terminal protein), spacer, RT (Reverse transcriptase) and RNaseH. Among these domains, tyrosine, the 65th residue of TP is an important residue in protein-priming reaction that initiates reverse transcription. If monoclonal antibody that recognizes around tyrosine residue were selected, it could be applied to further study of HBV replication. METHODS: To produce TP-specific scFv (single-chain Fv) by phage display, mice were immunized using synthetic TP-peptide contains 57~80th amino acid residues of TP domain. After isolation of mRNA of heavy-variable region (VH) and light-chain variable region (VL) from the spleen of the immunized mouse, DNA of VH and VL were obtained by RT-PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a scFv DNA fragments. ScFv DNA fragments were cloned into a phagemid vector. ScFv was expressed in E.coli TG1 as a fusion protein with E tag and phage gIII. To select the scFv that has specific affinity to TP-peptide from the phage-antibody library, we used two cycles of panning and colony lift assay. RESULTS: The TP-peptide-specific scFv was isolated by selection process using TP-peptide as an antigen. Selected scFv had 30 kDa of protein size and its nucleotide sequences were analyzed. Indirect- and competitive-ELISA revealed that the selected scFv specifically recognized both TP-peptide and the HBV DNA polymerase. CONCLUSION: The scFv that recognizes the TP domain of the HBV DNA polymerase was isolated by phage display.