Standardized cotton swab sampling with nested quantitative polymerase chain reaction is effective for diagnosing ordinary scabies.

BACKGROUND: Low sensitivity of the PCR assay for diagnosing scabies has been noted because of the difficulty in obtaining tissue containing Sarcoptes scabiei DNA. AIM: To evaluate nested real-time quantitative PCR (nRT-qPCR) with nonexpert-dependent standardized cotton swab sampling (CSW) as a tool for diagnosing scabies. METHODS: All patients underwent dermoscopic and microscopic examination (MS) with scraped sampling (Sc). Patient samples were acquired with a single, dry swab rubbed across the flexor areas of both wrists as well as the eight interdigital spaces and on any suspected scabies lesions. nRT-qPCRs were performed with Sc and CSW samples. RESULTS: Out of 125 patients with suspected scabies, 120 patients were sampled, and 57 were positive (positive with: MS n = 53; nRT-qPCR with Sc n = 52; nRT-qPCR with CSW n = 46) and 63 were negative for scabies. The sensitivities of these tests were 93.0%, 91.2% and 80.7%, respectively, which were not different statistically (P > 0.05). However, upon subsequent monitoring after treatment, the sensitivity of nRT-qPCR with CSW was only 36.6%, which was significantly lower than 83.0% for MS and 92.7% for nRT-qPCR with Sc (P < 0.001). The obtained sequences showed 97%-100% homology with scabies sequences deposited in GenBank. CONCLUSION: CSW with nRT-qPCR shows sensitivity close to MS with scraping performed by experts for diagnosing scabies in an outpatient setting, but not for post-treatment monitoring. CSW with nRT-qPCR may be useful for physicians unfamiliar with a traditional diagnostic method, and for screening an outbreak in community facilities.

Cohnella cholangitidis sp. nov., a novel species of the genus Cohnella isolated from a clinical specimen in Korea.

A Gram-positive, aerobic, rod-shaped bacterium, designated as strain 1605-214T, was isolated from the blood sample of a patient with cholangitis. Based on its 16S rRNA gene sequence, the strain 1605-214T belonged to the genus Cohnella and exhibited 97.9% sequence identity with Cohnella luojiensis DSM 24270T (GQ214052). DNA-DNA hybridization, digital DNA-DNA hybridization, and average nucleotide identity values between the two species were 23% ± 1.9, 21.1%, and 77.2%, respectively. The cellular fatty acids of strain 1605-214T were mainly comprised of anteiso-C15:0 (36.1%), iso-C16:0 (16.5%), and C16:0 (15.1%). The predominant quinone was menaquinone-7; predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, and aminophospholipid-1. The cell wall peptidoglycan of strain 1605-214T contained meso-diaminopimelic acid. DNA G + C content of strain 1605-214T was 50.6 mol%. 5187 genes out of a total of 5413 (94.6%) were assigned putative functions using eggNOG v5.0. Based on genotypic characteristics and genomic sequence analysis results, strain 1605-214T was confirmed to represent a novel species of genus Cohnella, for which the name Cohnella cholangitidis sp. nov., was proposed.

Complexation of drug and hapten-conjugated aptamer with universal hapten antibody for pancreatic cancer treatment.

Owing to a lack of reliable markers and therapeutic targets, pancreatic ductal adenocarcinoma (PDAC) remains the most lethal malignant tumor despite numerous therapeutic advances. In this study, we utilized cell-SELEX to isolate a DNA aptamer recognizing the natural conformation of the target on the cell surface. PAp7T8, an aptamer optimized by size and chemical modification, exhibited specific targeting to pancreatic cancer cells and orthotopic xenograft pancreatic tumors. To confer therapeutic functions to the aptamer, we adopted a drug-conjugated oligobody (DOligobody) strategy. Monomethyl auristatin E was used as a cytotoxic drug, digoxigenin acted as a hapten, and the humanized anti-digoxigenin antibody served as a universal carrier of the aptamer. The resulting PAp7T8-DOligobody showed extended in vivo half-life and markedly inhibited tumor growth in an orthotopic pancreatic cancer xenograft model without causing significant toxicity. Therefore, PAp7T8-DOligobody represents a promising novel therapeutic delivery platform for PDAC.

Effect of GCSB-5, a Herbal Formulation, on Monosodium Iodoacetate-Induced Osteoarthritis in Rats.

Therapeutic effects of GCSB-5 on osteoarthritis were measured by the amount of glycosaminoglycan in rabbit articular cartilage explants in vitro, in experimental osteoarthritis induced by intra-articular injection of monoiodoacetate in rats in vivo. GCSB-5 was orally administered for 28 days. In vitro, GCSB-5 inhibited proteoglycan degradation. GCSB-5 significantly suppressed the histological changes in monoiodoacetate-induced osteoarthritis. Matrix metalloproteinase (MMP) activity, as well as, the levels of serum tumor necrosis factor-α, cyclooxygenase-2, inducible nitric oxide synthase protein, and mRNA expressions were attenuated by GCSB-5, whereas the level of interleukin-10 was potentiated. By GCSB-5, the level of nuclear factor-κB p65 protein expression was significantly attenuated but, on the other hand, the level of inhibitor of κB-α protein expression was increased. These results indicate that GCSB-5 is a potential therapeutic agent for the protection of articular cartilage against progression of osteoarthritis through inhibition of MMPs activity, inflammatory mediators, and NF-κB activation.

Establishment of Patient-derived Preclinical Models for Invasive Papillary Cholangiocarcinoma.

BACKGROUND/AIM: Invasive papillary cholangio-carcinoma (IPC) is a minor subtype of extrahepatic cholangiocarcinoma. However, its etiology and characteristics remain unknown because of the unavailability of in vitro and in vivo models. We aimed to establish a novel preclinical model for translational research of IPC. MATERIALS AND METHODS: A patient-derived xenograft (PDX) was engrafted in NOG mice and the cell line National Cancer Center human IPC (NCChIPC) was subsequently established from the PDX tumors. Immunohistochemistry and RNA-sequencing were used to determine the retention of original characteristics of patient tissues. RESULTS: PDX tumors showed successful amplification, and the NCChIPC-derived xenograft largely retained the histopathological features of the original tumor with CK19, MUC1 and MUC5AC expression. Transcriptome analysis showed a high correlation between patient and preclinical models. Additionally, anticancer drugs response was analyzed in the NCChIPC PDX. CONCLUSION: These novel preclinical models here will help elucidate IPC etiology and facilitate translational research.

Hepatoprotective effect of pinoresinol on carbon tetrachloride-induced hepatic damage in mice.

Forsythiae Fructus is known to have diuretic, anti-bacterial, and anti-inflammatory activities. This study examined the hepatoprotective effects of pinoresinol, a lignan isolated from Forsythiae Fructus, against carbon tetrachloride (CCl(4))-induced liver injury. Mice were treated intraperitoneally with vehicle or pinoresinol (25, 50, 100, and 200 mg/kg) 30 min before and 2 h after CCl4 (20 microl/kg) injection. In the vehicle-treated CCl(4 )group, serum aminotransferase activities were significantly increased 24 h after CCl4 injection, and these increases were attenuated by pinoresinol at all doses. Hepatic glutathione contents were significantly decreased and lipid peroxidation was increased after CCl4 treatment. These changes were attenuated by 50 and 100 mg/kg of pinoresinol. The levels of protein and mRNA expression of inflammatory mediators, including tumor necrosis factor-alpha, inducible nitric oxide synthase, and cyclooxygenase-2, were significantly increased after CCl4 injection; and these increases were attenuated by pinoresinol. Nuclear translocation of nuclear factor-kappaB (NF-kappaB) and phosphorylation of c-Jun, one of the components of activating protein 1 (AP-1), were inhibited by pinoresinol. Our results suggest that pinoresinol ameliorates CCl4)-induced acute liver injury, and this protection is likely due to anti-oxidative activity and down-regulation of inflammatory mediators through inhibition of NF-kappaB and AP-1.

Sleep disturbance in women with eating disorder: prevalence and clinical characteristics.

The prevalence of sleep disturbance among 400 patients, diagnosed with anorexia nervosa (AN) or bulimia nervosa (BN), was assessed via structured interviews. Sleep disturbance was reported in 50.3% of patients, especially among those with the binge-eating/purging subtype. Patients with sleep disturbance had more disturbing symptoms; including higher binge frequency and vomiting frequency. Additional differences were also identified.

Protective effects of human placenta extract on cartilage degradation in experimental osteoarthritis.

This study investigated the effect of human placenta extract (HPE) on cartilage degradation in vitro MG-63 cells, articular cartilage explants, and in vivo monoiodoacetate (MIA)-induced osteoarthritis (OA). Matrix metalloproteinase (MMP)-2 activity was measured in HPE-treated osteoblastic MG-63 cells. Articular cartilage explants in rabbit were cultured, and the degree of proteoglycan (PG) degradation was assessed by measuring the amount of glycosaminoglycan (GAG) released into the culture medium. Experimental osteoarthritis was induced by intra-articular injection of 3 mg MIA in rats. Beginning 14 d post-MIA injection, HPE was administered intra-articularly once a day for 14 d. The knee joints were assessed by roentgenography, histology, and gelatinase activity. HPE inhibited PG degradation in articular cartilage explants. HPE significantly reduced deformity of knee joints and suppressed the histological change in MIA-induced OA. HPE inhibited MMP-2 activity in MG-63 cells. MMP-2 and -9 activities were also reduced in the cartilages of HPE-treated knee joints. Our results indicate that HPE has therapeutic effects on OA by protecting cartilage.

Intradiploic epidermoid cyst in a dog.

CASE DESCRIPTION: A 10-year-old 7-kg (15.4-lb) neutered male Shih Tzu was referred for evaluation because of a sudden onset of dullness and intermittent vomiting of 1 to 2 weeks' duration. Two days prior to evaluation, clinical signs had worsened and 1 seizure was reported. CLINICAL FINDINGS: A 3.5 × 2.5-cm soft, dome-shaped mass of the right occipital region of the head was noted on physical examination. Radiography and CT confirmed the presence of the mass and indicated its intra- and extracranial expansion. The MRI images showed compression of the cerebellum by the mass, with distinct margins of hypointensity on both T1- and T2-weighted images. TREATMENT AND OUTCOME: Surgery was performed, and the mass was completely excised. The dog had mild ataxia, hypermetria, and head tremors after surgery. Histologic examination of the mass yielded a diagnosis of intradiploic epidermoid cyst. On examination 3 weeks after surgery, the previous neurologic signs had resolved. On examination 25 months after surgery, the dog remained free of clinical signs. CLINICAL RELEVANCE: The present report described the clinical signs, diagnostic imaging results, and successful surgical removal of an intradiploic epidermoid cyst in a dog. Long-term prognosis may be good with complete removal of intradiploic epidermoid cysts in dogs.

A Variant of SLC1A5 Is a Mitochondrial Glutamine Transporter for Metabolic Reprogramming in Cancer Cells.

Glutamine is an essential nutrient that regulates energy production, redox homeostasis, and signaling in cancer cells. Despite the importance of glutamine in mitochondrial metabolism, the mitochondrial glutamine transporter has long been unknown. Here, we show that the SLC1A5 variant plays a critical role in cancer metabolic reprogramming by transporting glutamine into mitochondria. The SLC1A5 variant has an N-terminal targeting signal for mitochondrial localization. Hypoxia-induced gene expression of the SLC1A5 variant is mediated by HIF-2α. Overexpression of the SLC1A5 variant mediates glutamine-induced ATP production and glutathione synthesis and confers gemcitabine resistance to pancreatic cancer cells. SLC1A5 variant knockdown and overexpression alter cancer cell and tumor growth, supporting an oncogenic role. This work demonstrates that the SLC1A5 variant is a mitochondrial glutamine transporter for cancer metabolic reprogramming.

Measurement of Vibrating Tympanic Membrane in an In Vivo Mouse Model Using Doppler Optical Coherence Tomography.

Optical coherence tomography (OCT) has a micro-resolution with a penetration depth of about 2 mm and field of view of about 10 mm. This makes OCT well suited for analyzing the anatomical and internal structural assessment of the middle ear. To study the vibratory motion of the tympanic membrane (TM) and its internal structure, we developed a phase-resolved Doppler OCT system using Kasai's autocorrelation algorithm. Doppler optical coherence tomography is a powerful imaging tool which can offer the micro-vibratory measurement of the tympanic membrane and obtain the micrometer-resolved cross-sectional images of the sample in real-time. To observe the relative vibratory motion of individual sections (malleus, thick regions, and the thin regions of the tympanic membrane) of the tympanic membrane in respect to auditory signals, we designed an experimental study for measuring the difference in Doppler phase shift for frequencies varying from 1 to 8 kHz which were given as external stimuli to the middle ear of a small animal model. Malleus is the very first interconnecting region between the TM and cochlea. In our proposed study, we observed that the maximum change in Doppler phase shift was seen for the 4 kHz acoustic stimulus in the malleus, the thick regions, and in the thin regions of the tympanic membrane. In particular, the vibration signals were higher in the malleus in comparison to the tympanic membrane.

Detection of Gram-negative bacterial outer membrane vesicles using DNA aptamers.

Infection of various pathogenic bacteria causes severe illness to human beings. Despite the research advances, current identification tools still exhibit limitations in detecting Gram-negative bacteria with high accuracy. In this study, we isolated single-stranded DNA aptamers against multiple Gram-negative bacterial species using Toggle-cell-SELEX (systemic evolution of ligands by exponential enrichment) and constructed an aptamer-based detection tool towards bacterial secretory cargo released from outer membranes of Gram-negative bacteria. Three Gram-negative bacteria, Escherichia coli DH5α, E. coli K12, and Serratia marcescens, were sequentially incubated with the pool of random DNA sequences at each SELEX loop. Two aptamers selected, GN6 and GN12, were 4.2-times and 3.6-times higher binding to 108 cells of Gram-negative bacteria than to Gram-positive bacteria tested, respectively. Using GN6 aptamer, we constructed an Enzyme-linked aptamer assay (ELAA) to detect bacterial outer membrane vesicles (OMVs) of Gram-negative bacteria, which contain several outer membrane proteins with potent immunostimulatory effects. The GN6-ELAA showed high sensitivity to detect as low as 25 ng/mL bacterial OMVs. Aptamers developed in this study show a great potential to facilitate medical diagnosis and early detection of bacterial terrorism, based on the ability to detect bacterial OMVs of multiple Gram-negative bacteria.

Acid-base chemistry at the ice surface: reverse correlation between intrinsic basicity and proton-transfer efficiency to ammonia and methyl amines.

Proton transfer from the hydronium ion to NH(3), CH3NH2, and (CH3)2NH is examined at the surface of ice films at 60 K. The reactants and products are quantitatively monitored by the techniques of Cs+ reactive-ion scattering and low-energy sputtering. The proton-transfer reactions at the ice surface proceed only to a limited extent. The proton-transfer efficiency exhibits the order NH3>(CH3)NH2=(CH3)2NH, which opposes the basicity order of the amines in the gas phase or aqueous solution. Thermochemical analysis suggests that the energetics of the proton-transfer reaction is greatly altered at the ice surface from that in liquid water due to limited hydration. Water molecules constrained at the ice surface amplify the methyl substitution effect on the hydration efficiency of the amines and reverse the order of their proton-accepting abilities.

Bicyclist injury severities in bicycle-motor vehicle accidents.

This research explores the factors contributing to the injury severity of bicyclists in bicycle-motor vehicle accidents using a multinomial logit model. The model predicts the probability of four injury severity outcomes: fatal, incapacitating, non-incapacitating, and possible or no injury. The analysis is based on police-reported accident data between 1997 and 2002 from North Carolina, USA. The results show several factors which more than double the probability of a bicyclist suffering a fatal injury in an accident, all other things being kept constant. Notably, inclement weather, darkness with no streetlights, a.m. peak (06:00 a.m. to 09:59 a.m.), head-on collision, speeding-involved, vehicle speeds above 48.3 km/h (30 mph), truck involved, intoxicated driver, bicyclist age 55 or over, and intoxicated bicyclist. The largest effect is caused when estimated vehicle speed prior to impact is greater than 80.5 km/h (50 mph), where the probability of fatal injury increases more than 16-fold. Speed also shows a threshold effect at 32.2 km/h (20 mph), which supports the commonly used 30km/h speed limit in residential neighborhoods. The results also imply that bicyclist fault is more closely correlated with greater bicyclist injury severity than driver fault.

Spatial regression analysis of traffic crashes in Seoul.

Traffic crashes can be spatially correlated events and the analysis of the distribution of traffic crash frequency requires evaluation of parameters that reflect spatial properties and correlation. Typically this spatial aspect of crash data is not used in everyday practice by planning agencies and this contributes to a gap between research and practice. A database of traffic crashes in Seoul, Korea, in 2010 was developed at the traffic analysis zone (TAZ) level with a number of GIS developed spatial variables. Practical spatial models using available software were estimated. The spatial error model was determined to be better than the spatial lag model and an ordinary least squares baseline regression. A geographically weighted regression model provided useful insights about localization of effects. The results found that an increased length of roads with speed limit below 30 km/h and a higher ratio of residents below age of 15 were correlated with lower traffic crash frequency, while a higher ratio of residents who moved to the TAZ, more vehicle-kilometers traveled, and a greater number of access points with speed limit difference between side roads and mainline above 30 km/h all increased the number of traffic crashes. This suggests, for example, that better control or design for merging lower speed roads with higher speed roads is important. A key result is that the length of bus-only center lanes had the largest effect on increasing traffic crashes. This is important as bus-only center lanes with bus stop islands have been increasingly used to improve transit times. Hence the potential negative safety impacts of such systems need to be studied further and mitigated through improved design of pedestrian access to center bus stop islands.