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Background Behçet’s disease (BD) is a systemic inflammatory vasculitis with both autoimmune and autoinflammatory properties. Renal involvement in BD and its spontaneous remission have been rare. We herein describe a case of parallel disease activity of BD with entero and renal involvements, followed by a spontaneous remission without corticosteroid treatment. Case presentation A 54-year-old woman who had a 4-year history of BD, maintained with colchicine treatment, suffered abdominal pain, hemorrhagic stool and diarrhea. Physical examination revealed strong tenderness in the entire abdomen. Laboratory test results showed increased levels of inflammation, and a computed tomography scan revealed edematous intestinal wall thickening with ascites. Blood and stool cultures showed no specific findings. Since she was suspected to have developed panperitonitis with acute enterocolitis, she started treatment with an antibacterial agent under bowel rest. Her abdominal symptoms gradually improved, while diarrhea and high levels of inflammatory reaction persisted. Colonoscopy revealed discontinuous abnormal mucosal vascular patterns and ulcerations in the whole colon except for the rectum, and histological analyses of the intestine demonstrated transmural mucosal infiltration of inflammatory cells without epithelioid granuloma or amyloid deposition. Based on these findings, she was diagnosed with entero BD. Meanwhile, pedal edema appeared during her hospitalization. Urinalysis results were consistent with nephrotic syndrome, thus a renal biopsy was performed. Light microscopy showed no obvious glomerular and interstitial abnormalities, whereas electron microscopy revealed foot process effacement without immune complex deposition or fibrillary structure, compatible with minimal change disease (MCD). Only with conservative therapy, her proteinuria decreased, followed by a complete remission in 3 weeks from the onset of edema. The coincident episode of MCD was finally diagnosed as renal BD that paralleled disease activity to entero BD. She started adalimumab administration, resulting in the further improvement of diarrhea and inflammatory levels. Conclusions This is the first report to demonstrate MCD as renal involvement of BD along with the disease activity of entero BD.

In this paper, reinforcement of dielectric nanocomposite was performed for polyvinyl fluoride (PVDF) as a matrix, which is a practical requirement for foldable and flexible electric energy devices. A fibrous nanofiller, 2,2,6,6-Tetramethylpiperidine 1-oxyl (TEMPO)-oxidized cellulose nanofiber (TOCN) was employed as a nanofiller for the PVDF nanocomposite incorporating highly dielectric barium titanate nanoparticles (BT NPs). Prior to the fabrication of nanocomposite films, a chemical treatment by HCl and a surface-modification with trifluoropropyl group were applied to TOCN and BT nanoparticles, respectively. Their treatments brought about both nanofillers well-dispersed in a polar aprotic solvent for PVDF. A low percentage incorporation of the treated TOCNs up to 0.5 wt% was found to be sufficient for enhancement on the mechanical strength of nanocomposites at a high content of surface-modified BT NPs (30 vol%). The permittivity of the nanocomposites was in the range of 12 to 14 that is twice higher than that in the PVDF film. The PVDF nanocomposites at the treated TOCN loading of 0.3 to 0.5 wt% exhibited superior mechanical strength to the PVDF film. The ternary combination of PVDF, the treated TOCN, and the surface-modified BT NPs showed an improvement on tensile stiffness with maintaining the intrinsic high dielectric properties of nanocomposite films. The present nanocomposite can be a promising ternary system for an advanced design of polymer dielectrics with excellent energy storage properties.

Thrombomodulin (TM), which is predominantly expressed on the endothelium, plays an important role in maintaining vascular homeostasis by regulating the coagulation system. Intravascular injury and inflammation are complicated physiological processes that are induced by injured endothelium-mediated pro-coagulant signaling, necrotic endothelial- and blood cell-derived damage-associated molecular patterns (DAMPs), and DAMP-mediated inflammation. During the hypercoagulable state after endothelial injury, TM is released into the intravascular space by proteolytic cleavage of the endothelium component. Recombinant TM (rTM) is clinically applied to patients with disseminated intravascular coagulation, resulting in protection from tissue injury. Recent studies have revealed that rTM functions as an inflammatory regulator beyond hemostasis through various molecular mechanisms. More specifically, rTM neutralizes DAMPs, including histones and high mobility group box 1 (HMGB1), suppresses excessive activation of the complement system, physiologically protects the endothelium, and influences both innate and acquired immunity. Neutrophil extracellular traps (NETs) promote immunothrombosis by orchestrating platelets to enclose infectious invaders as part of the innate immune system, but excessive immunothrombosis can cause intravascular injury. However, rTM can directly and indirectly regulate NET formation. Furthermore, rTM interacts with mediators of acquired immunity to resolve vascular inflammation. So far, rTM has shown good efficacy in suppressing inflammation in various experimental models, including thrombotic microangiopathy, sterile inflammatory disorders, autoimmune diseases, and sepsis. Thus, rTM has the potential to become a novel tool to regulate intravascular injury via pleiotropic effects.

Objective: ANCA associated vasculitis (AAV) is characterized by systemic necrotizing vasculitis with the presence of ANCA. Although BPI-ANCA is one of the atypical ANCAs and is occasionally seen in patients with vasculitis, the pathogenicity of BPI-ANCA remains unclear. This study was performed to examine the pathogenic role of BPI-ANCA against neutrophils. Methods: A 76-year-old Japanese man showed BPI-ANCA positive systemic vasculitis with a medical history of Pseudomonas aeruginosa infection. BPI-ANCA IgGs were eluted from the patient serum using an immunoadsorbent column. In vitro experiment, healthy donor neutrophils were treated with BPI-AAV IgGs, MPO-AAV IgGs, healthy control IgGs under TNFα stimulation. After 3 h incubation, neutrophil extracellular trap (NET) was assessed by immunofluorescent imaging. To determine the pathogenicity of BPI-ANCA, TNFα-primed neutrophils were incubated with monoclonal BPI-ANCA in the presence or absence of recombinant BPI. Results: BPI-AAV IgGs-treated neutrophils showed NET formation with histone citrullination. Interestingly, the monoclonal BPI-ANCA did not induce NET, but the immune complexes (ICs) of recombinant BPI and BPI-ANCA induced TNFα-dependent NET formation with hypercitrullination. Furthermore, TNFα increased the expression of BPIs in neutrophils and the BPIs were translocated to cell surface. Conclusion: BPI-ANCA could affect neutrophils leading to NET formation and may play a role in the development of systemic vasculitis as pathogenic autoantibody.

Nephrotic syndrome without hematuria due to infection-related glomerulonephritis is uncommon. The present report describes a case of nephrotic syndrome due to infection-related glomerulonephritis without hematuria and hypertension in an older child. A 14-year-old boy was referred to our hospital because of a 5-day history of fever, nausea, weight gain and recent leg edema without hypertension. Laboratory data showed nephrotic-range proteinuria, hypoalbuminemia, mild hypocomplementemia and acute renal injury without hematuria. Although, due to the clinical presentation, minimal-change nephrotic syndrome was mostly suspected, a renal biopsy showed endocapillary hypercellularity mainly of mononuclear cells with segmental mesangiolytic changes. Fine granular IgG and C3 deposits were noted by an immunofluorescent study; many relatively small electron-dense deposits were observed electron-microscopically. These findings led to the diagnosis of nephrotic syndrome due to infection-related endocapillary proliferative glomerulonephritis, although the causative organism of his nephritis was not detected. He recovered with rest and dietary cure. When we examine an acute nephrotic child, infection-related glomerulonephritis should be considered as the differential diagnosis to avoid unnecessary use of corticosteroids.

An autopsy for a suicidal case of a male in his 40s, who had died of poisoning due to ingestion of a large amount of buformin, was performed at our department. Buformin is biganide class agent used for patients of diabetes mellitus, which can occasionally cause severe lactic acidosis. The autopsy was performed about 10 days after his death, and the direct cause of his death was judged as asphyxia due to the aspiration of stomach contents into the airway. The nine body fluids and eight solid tissues specimens were dealt with for investigating postmortem distribution/redistribution of buformin in a whole body; femoral vein blood, right and left heart blood, pericardial fluid, urine, bile, stomach contents, small intestine contents, cerebrospinal fluid, the brain, lung, heart muscle, liver, spleen, kidney and skeletal muscle were examined. For extracting buformin from specimens, a modified QuEChERS method including dispersive solid-phase extraction was employed, followed by the analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS). Buformin in various kinds of human matrices were quantified by the standard addition method in this study, which can overcome the matrix effects and recovery rates without use of blank human matrices. All concentrations of buformin in specimens examined in this case were extremely higher than those of previously reported poisoning cases. The concentrations of buformin in left and right heart blood and femoral vein blood specimens of this case were 399, 216 and 261μg/mL, respectively; although the direct cause of his death was judged as asphyxia due to occlusion of airway with stomach contents, the vomiting was thought to be provoked by buformin poisoning. In this study, marked differences of buformin concentrations between brain tissue and cerebral spiral fluids, and other specimens were observed, which suggested that its distribution was influenced also by blood-brain-barrier. Although a number of buformin poisoning cases were published so far, they gave sporadic data on its concentrations and/or distribution in some limited human specimens. This study is the first to describe detailed distribution/redistribution of buformin in a whole human body quantified by using LC-MS/MS.

A method to prevent sintering of photocatalytic particles in a calcination process is developed to allow their crystallization without a significant decrease in their specific surface area. A sacrificial layer of polymer to prevent the particle from sintering was employed for the crystallization of amorphous titania particles in the heat treatment at 500 °C. The heat treatment simultaneously removed the polymer layer by pyrolysis. The heat treatment using a sacrificial layer of polymer could suppress a decrease in the specific surface area of titania particles and also maintain colloidal stability in water, attaining a photocatalytic activity higher than that without using a sacrificial layer. These results indicate that the polymer-coating technique is effective for crystallization of photocatalytic particles without a loss of both colloidal stability and photocatalytic activity of the particles.

Background Enterovirus D68 (EV-D68) is a re-emerging, high-morbidity pathogen that causes severe respiratory infection and acute flaccid myelitis in children. EV-D68 is phylogenetically divided into 4 major groups—clade A through D—and each clade has subclades defined by genetic variant. Outbreaks of all strains have been reported in more than 10 countries. However, no study has compared the viral genomic characteristics of EV-D68 in contemporaneous outbreaks in different countries. Methods An outbreak of EV-D68 in children younger than 15 years occurred in Niigata, Japan, from October through November 2018. The patients were admitted to hospital with respiratory distress and wheezing episodes. RNA extracted from nasopharyngeal samples was tested by EV-D68-specific PCR. Clade was determined by semi-nested PCR analysis of the VP1 region, and the phylogenetic tree of the VP1 sequence was constructed. To clarify viral genomic characteristics, we compared the clade to those of outbreaks occurring during 2014–2018 in other countries. Results EV-D68 testing of 47 patients yielded positive results for 22 (47%) (median age 4.6 years; IQR, 2.9–6.7), and 15 (69%) had a past medical history of allergic disease. No patient developed acute flaccid myelitis. The VP1 sequences from all isolates belonged to clade B3, the same clade detected during the 2015 outbreak in Japan (figure). Interestingly, EV-D68 outbreaks were reported in France and Italy (clades D1/B3 and D1, respectively) during this period. Although the EV-D68 clades responsible for outbreaks in 2014 differed by region, clade B3 is now the dominant clade and has caused concurrent EV-D68 outbreaks in children since 2015 (table). Conclusion In 2018, an EV-D68 clade B3 outbreak occurred among children in Niigata, Japan. This clade is now dominant and periodically circulates around the world. Because EV-D68 can cause simultaneous outbreaks worldwide and is associated with high morbidity, detailed real-time epidemiological surveillance of EV-D68 is warranted. Disclosures All authors: No reported disclosures.

Mammalian pluripotent stem cells possess properties of self-renewal and pluripotency. These abilities are maintained by the strict regulation of pluripotent stem cell-specific transcription factor network and unique properties of chromatin in the stem cells. Although these major signaling pathways robustly control the characteristics of stem cells, other regulatory factors, such as metabolic pathways, are also known to modulate stem cell proliferation and differentiation. In this study, we fractionated protein samples from mouse embryonic stem (ES) cells cultured with or without the leukemia inhibitory factor (LIF). Protein expression was quantified by 2-dimensional differential gel electrophoresis (2D-DIGE). In total, 44 proteins were identified as being differentially expressed in the pluripotent stem cells and the differentiated cells. Surprisingly, half of the identified proteins were the proteins localized in mitochondria, which supply cellular energy and regulate cell cycle, development, and cell death. Some of these identified proteins are involved in the metabolic function and the regulation of pluripotency. Further analysis of the identified proteins could provide new information for the manipulation of pluripotency in ES cells.

The chemically-induced isomerization of stilbenes during the peroxyoxalate chemiluminescence (PO-CL) reactions was reinvestigated. The PO-CL reactions using bis(2,4,6-trichlorophenyl) oxalate in the presence of several stilbenes (type A reaction) produced cis-stilbenes in 0-4% yields, which was dependent on the singlet excitation energy of the stilbenes. On the other hand, the PO-CL reactions of the oxalates, containing the stilbene moieties in the molecules (type B reaction), produced cis-stilbenes 0-9.3% yields, some of which were much more effective than the type A reactions considering the amount of the oxalate moiety as the energy supplier., Article, TETRAHEDRON LETTERS. 55(3):619-622 (2014)

A rapid and sensitive detection method using matrix-assisted laser desorption ionization (MALDI) — time-of-flight (TOF) — mass spectrometry (MS) was developed for the analysis of four pyrrolidino cathinones, such as α-pyrrolidinopropiophenone (PPP), 4’-methyl-α-pyrrolidinopropiophenone (MPPP), α-pyrrolidinobutiophenone (PBP), and α-pyrrolidinovalerophenone (PVP). In this method, α-cyano-4-hydroxy cinnamic acid was used as the matrix to assist the ionization of cathinones. Each MALDI-TOF-MS spectrum of these cathinones showed not only protonated molecular ion [M + H]+, but also several fragment ions having comparable intensities to that of [M + H]+. Hence MPPP and PBP could be clearly discriminated by the mass spectra alone, although they have almost the same mass numbers in their protonated molecular ions. The quantification of MPPP, PBP or PVP was performed using PPP as internal standard, respectively, and that of PPP was performed using PBP as internal standard. The limit of detection was 1 ng/ml, and the quantification range was 2 – 200 ng/ml for the four cathinones using 20 μl of blood. In a fatal poisoning case in which PVP was abused, the PVP levels in whole blood samples obtained from the right heart, left heart and femoral vein were 0.597, 0.635 and 0.580 μg/ml, respectively. We recommend the MALDI-TOF-MS method without any chromatography for both identification and quantification of the pyrrolidino cathinones in various matrices in forensic toxicological analysis, because of the simplicity, rapidness and reliability of the method.

Background Parechovirus-A3 (PeV-A3) is an emerging pathogen causing sepsis and meningoencephalitis in neonates and young infants. We previously reported that maternal antibodies against PeV-A3 are important to protect neonates and young infants from the infection. Recent studies showed that (1) breastfeeding had a protective effect against enterovirus, which is closely-related virus to PeV-A, and (2) human breast milk (HBM) neutralized enterovirus in vitro. Currently, no report is available related to the antiviral effect of HBM against PeV-A3. Methods HBM (colostrum, 3–5 days after childbirth; mature milk, 1 month after childbirth) and serum (within ± 1 week of child’s birthday) samples were obtained from mothers at obstetrics clinic in Niigata, Japan. Neutralizing antibody titers (NATs) against PeV-A3 were measured using the Vero cells. Results The anti-PeV-A3 NATs of colostrum (n = 32) ranged from 1:8 to 1:2048, those ≥1:32 was 59% (19/32). Whereas, the anti-PeV-A3 NATs of mature milk ranged from 1:8 to 1:96. and those ≥1:32 was 20% (2/20) (P < 0.001). The median NATs anti-PeV-A3 was higher in colostrum (1:32) compared with mature milk (1:8) (P < 0.001). There was a strong positive correlation between the NATs of colostrum and serum (r = 0.604, P < 0.001, Figure). Conclusion This study showed that HBM had high NATs against PeV-A3, which was correlated with serum NATs. Further studies are necessary to investigate which components of HBM has antiviral effects against PeV-A3. Disclosures All authors: No reported disclosures.

A rapid screening method was developed for the determination of phenothiazines by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF–MS). In this method, α-cyano-4-hydroxy cinnamic acid (CHCA) was used as the matrix to assist the ionization of phenothiazines. The identification of 11 phenothiazines with heavy side chains was achieved by observing their protonated molecular ions [M+H]+ at m/z 340–447. Quantification was achieved by using triflupromazine at m/z 353 as the internal standard (IS). The relative ionization efficiencies of 11 phenothiazines and IS on MALDI-TOF–MS were different from those achieved by ESI-MS, but the product ion spectra from MALDI-MS–MS were quite similar to those from ESI-MS–MS except in the case of flupentixol. The limit of detection was 0.3 ng/ml with a quantification range of 1–50 ng/ml urine in the best case; the limit of detection was 5 ng/ml with a quantification range of 10–100 ng/ml urine in the worst case for 10 phenothiazines except thiethylperazine. The present method provides a simple and high-throughput method for the screening of these phenothiazines using only 20 μl of urine. To our knowledge, this study is the first trial to analyze phenothiazines by MALDI-TOF–MS (–MS).

A rapid and sensitive method was developed for simultaneous determination of perchlorate (ClO4 −), thiocyanate (SCN−), and iodide (I−). The ions were reacted with tris(1,10-phenanthroline)iron(II) chelating cation, [Fe(phen)3]2+, to produce their ternary complexes, [Fe(phen)2ClO4]+, [Fe(phen)2SCN]+, and [Fe(phen)2I]+ that were extracted together with nitrobenzene. Five microliters of the extract was injected directly into an electrospray ionization tandem mass spectrometer. Quantification of ClO4 −, SCN−, and I− was performed by selected reaction monitoring of the product ion [Fe(phen)O2]+ at m/z 268 that derived from the precursor ion [Fe(phen)2ClO4]+ at m/z 515, the product ion [Fe(phen)SCN]+ at m/z 294 that derived from precursor ion [Fe(phen)2SCN]+ at m/z 474, and the product ion [Fe(phen)I]+ at m/z 363 that derived from precursor ion [Fe(phen)2I]+ at m/z 543, respectively. Perchlorate, SCN−, and I− were measured in the range of 0.1–10 μM with the limits of detection at 0.03 μM by using a 50-μl sample aliquot containing the three ions within 10 min. The SCN− level in gastric fluid of a victim of NaCN poisoning was 137 ± 23 μM, which is slightly higher than the results from control subjects (n = 7), 72 ± 39 μM.

A very simple and sensitive method for the simultaneous analysis of naphthalene and p-dichlorobenzene in human whole blood and urine by headspace capillary gas chromatography-mass spectrometry (GC-MS) is presented. The advantages of the method were that as much as 1 mL of headspace vapor could be injected into a GC port in the splitless mode, and that the addition of deuterated naphthalene and p-dichlorobenzene as internal standards resulted in much better headspace extraction efficiencies, which resulted in high sensitivity. The detection limits for both naphthalene and p-dichlorobenzene were 1 ng mL(-1) for whole blood and 0.5 ng mL(-1) for urine. Validation data, such as the linearity of calibration curves, reproducibility and recovery rates, were all satisfactory. Using this method, both compounds could actually be detected from whole blood samples of a male volunteer after the inhalation of each gas of the compounds.

A high-performance liquid chromatography (HPLC) method has been developed for the assays of five barbiturates in human urine using a new thermoresponsive polymer separation column, which is composed of N-isopropylacrylamide polymer. According to elevating the column temperature from 10 ℃ to 50 ℃, five barbiturates, such as metharbital, primidone, phenobarbital, mephobarbital and pentobarbital, became well separated by this method. Five barbiturates showed good linearity in the range of 0.2-10 mg/ml. Good accuracy, precision and recoveries for these drugs were obtained at 1 and 5 μg/ml urine. The method with the new-type column seems to have high potential to be extensively used in forensic toxicology for analysis of many drugs and poisons by HPLC and HPLC-mass spectrometry (MS) (-MS).

A very simple method for the analysis of methamphetamine and amphetamine in human whole blood and urine by headspace gas chromatography (GC) has been presented. It neither needs solid-phase microextraction nor cryogenic trapping devices, but only a conventional capillary GC instrument with flame ionization detection (FID). The two special points to be mentioned in this method are the in-matrix derivatization of amphetamines for vaporization and the capability of injection of as large as 5 mL of the headspace vapor into a GC instrument in the splitless mode for sensitive detection. After heating a whole blood or urine sample containing amphetamines, α-methylbenzylamine (internal standard, IS) and heptafluoro-n-butyryl chloride under alkaline conditions in a 7.0-mL vial at 90 °C for 20 min, 5 mL of the headspace vapor was drawn with a glass syringe and injected into the gas chromatograph. During injection the column was at 40 °C to trap the analytes, and then the oven temperature was programmed up to 320 °C. Sharp peaks were obtained for each analyte and IS, and only a relatively small number of background impurity peaks for the whole blood and urine samples. The detection limits for each amphetamine were estimated to be 0.1 μg mL−1 for whole blood and 0.03 μg mL−1 for urine. Precision and linearity were also tested to confirm the reliability. Methamphetamine and amphetamine could be determined from whole blood and urine obtained at autopsy in three methamphetamine poisoning cases. The identity of each peak appearing in the gas chromatograms was confirmed by GC/mass spectrometry.

Background Human parechovirus (HPeV) type 3 and enteroviruses (EVs) are the common viruses causing severe diseases in neonates and young infants. Given no specific therapy is currently available and the morbidity and mortality of the diseases are high, understanding the detailed transmission routes is critical for the prevention. Methods This prospective study evaluated viral etiologies of neonates and young infants < 4 months suspecting sepsis and/or meningoencephalitis in 2016 in Niigata, Japan. DNA and RNA from serum/cerebrospinal fluid (CSF) were analyzed for HPeVs, EVs, and/or herpes simplex virus using real-time PCR. Bacterial infection was excluded based on blood/CSF culture results and their clinical course of the patients. To investigate the source of infection, we collected stool samples from available family members of the patients regardless of their symptoms. The stool samples were checked for each virus and if they were positive, the VP1 region for HPeVs and VP1/VP4-2 regions for EVs, were sequenced and typed. Furthermore, clinical information on symptomatic family members of HPeVs- and EVs-infected patients was compared. Results In total, 23/54 (43%) were positive for HPeVs (12/54, 22%) and EVs (11/54, 20%). Among the available family members for HPeVs and EVs, symptomatic family members were observed 28% (15/54) and 25% (13/53), and their stool samples were positive; 87% (13/15) and 100% (13/13), respectively. For available stool samples from asymptomatic family members, 36% (14/39) was positive for HPeV3 and 50% (20/40) were positive for EVs. Genetic sequence analyses confirmed the identical HPeVs and EVs; 92% (12/13) and 92% (12/13) in symptomatic patients, and 64% (9/14) and 70% (14/20) in asymptomatic patients, respectively. Rhinorrhea was the only symptom commonly observed in symptomatic family members infected with HPeVs (12/13, 92%) compared with those infected with EVs (4/13, 31%) (P = 0.004). Conclusion Symptomatic, and even asymptomatic children and adults can be a source of HPeVs- and EVs-infected neonates and young infants. Careful hand hygiene and other standard precaution may contribute to decrease the risk of the transmission during the outbreak of HPeV3 and EVs. Disclosures All authors: No reported disclosures.

Background Human parechovirus (HPeV) type 3 (HPeV3) is an emerging pathogen causing sepsis and meningoencephalitis in neonates and young infants. However, specific treatment for HPeV3 infection is currently unavailable. The application of antisense technology, such as peptide nucleic acids (PNAs), to viral infection has opened a new era of therapeutics. The aim of this study is to develop PNAs inhibiting HPeV3 gene expression in an in vitro model. Methods We designed four PNAs that target domains I, J (base and head of domain J structure), and K of an internal ribosomal entry site (IRES) region within the 5’ untranslated region of HPeV3. The IRES region is needed for the cap-independent translation. The PNAs were conjugated to cell-penetrating peptide (RXR)4XB (R = L-arginine, X = 6-aminohexanoic acid, B = β-alanine). LLC-MK2 cells were treated with 0.1–10µM of each PNA or water-containing growth medium for 4h. The cells were then infected with HPeV3 at the multiplicity of infection (MOI) of 10 for 1h. The infected cells were incubated for 7 days at 37ºC in 5% CO2. Extracellular levels of HPeV3 RNA were measured by real-time PCR on days 0 and 7. Results Without any treatment, an extracellular level of HPeV3 RNA increased to 8.2 × 106 copies/µL on day 7. When the cells were treated with 10µM of PNA targeting the domain I of IRES, an extracellular level of HPeV3 RNA was suppressed to 4.7 × 104 copies/µL (−99%) on day 7. Using the same PNA with lower concentrations, 1 µM and 0.1 µM of the PNA suppressed 24% and 0% of extracellular levels of HPeV3 RNA, respectively, which demonstrated the effect is dose-dependent. In contrast, 10µM of PNAs targeting domain J (base), J (head), and K suppressed 94%, 92%, and 20% of extracellular levels of HPeV3 RNA, respectively, compared with control. Conclusion The PNA-(RXR)4XB targeting the domain I of IRES suppressed extracellular levels of HPeV3 RNA in an in vitro model in a dose-dependent manner. Thus, PNA treatment may be a therapeutic candidate for HPeV3-infected patients. This novel therapy could target other HPeV genotypes given that the target sequence used in this study is identical to those of other clinically significant HPeVs. Disclosures All authors: No reported disclosures.

Background Human parechovirus type 3 (HPeV3) and enteroviruses (EVs) are the major viral pathogens causing severe diseases in neonates and young infants. Although two infections typically occur in summer, and initial clinical presentations are similar, differences in clinical and laboratory characteristics of HPeV3- and EV- infections are not well described. Methods From January 2014 to December 2016, we prospectively obtained serum and/or cerebrospinal fluid (CSF) samples from febrile neonates and young infants < 4 months suspecting sepsis and meningoencephalitis in Niigata, Japan. RNA/DNA were extracted from the samples and viral etiologies including HPeV3, EVs, and/or herpes simplex virus were tested using the real-time PCR, followed by genetic sequencing and typing. The comparison was made between HPeV3- and EVs-infected patients based on their medical records and subsequent questionnaires using appropriate statistical analyses. Results In total, we obtained 212 serum and 170 CSF samples from 222 patients. HPeV3 and EVs were detected for 56 (25%) and 43 patients (19%) (Figure), and their median ages were 29 and 24 days (P = 0.58), respectively. HPeV3-infected patients presented with higher median body temperature (38.9 °C vs 38.5 °C, P < 0.001), heart rates (184/minute vs. 161/minute, P = 0.002), peripheral coldness (72% vs. 34%, P < 0.001), skin mottling (65% vs. 23%, P < 0.001), and grunting (22% vs. 5%, P = 0.014) than EVs-infected patients. Additionally, systemic inflammatory response syndrome (SIRS) was more frequently observed in HPeV3-infected patients than in EVs-infected patients (82% vs. 58%, P = 0.009). In the laboratory data, median white blood cell count was lower in HPeV3-infected patients than those in EVs-infected patients (5,200/µL vs. 8,900/µL, P < 0.001). The pleocytosis was observed 58% of EVs-infected patients, but none of the HPeV3-infected patients (P < 0.001); however, HPeV3 RNA was frequently detected than EVs in CSF (90% vs 66%, P = 0.012). Conclusion This study showed that HPeV3-infected neonates and young infants presented more severe clinical manifestations than those infected with EVs. Significant tachycardia, poor peripheral circulation, and fulfilling SIRS criteria at the presentation may be the clues considering HPeV3 infection rather than EVs infection. Disclosures All authors: No reported disclosures.