bluuebunny/crossref_metadata_2025 huggingface dataset:
Women’s Views on Their Diagnosis and Management for Borderline Gestational Diabetes Mellitus
Authors: Han Shanshan , Middleton Philippa F. , Bubner Tanya K. , Crowther Caroline A.
Published: February 2015

Introduction. Little is known about women’s views relating to a diagnosis of borderline gestational diabetes mellitus (GDM) and the subsequent management. This study aimed to explore women’s experiences after being diagnosed with borderline GDM, their attitudes about treatment, and factors important to them for achieving any lifestyle changes.Methods. We conducted face-to-face, semistructured interviews with women diagnosed with borderline GDM.Results. A total of 22 women were interviewed. After a diagnosis of borderline GDM, 14 (64%) women reported not being concerned or worried. Management of borderline GDM was thought by 21 (95%) women to be very important or important. Eighteen (82%) women planned to improve their diet and/or exercise to manage their borderline GDM. The most frequently mentioned enabler for achieving intended lifestyle change was being more motivated to improve the health of their baby and/or themselves (15 women). The most frequent barrier was tiredness and/or being physically unwell (11 women).Conclusions. A diagnosis of borderline GDM caused some concern to one-third of women interviewed. The majority of women believed managing their borderline GDM was important and they planned to improve their lifestyle. Women’s own and their babies’ future health were powerful motivators for lifestyle change.
Frame Multiresolution Analysis on Local Fields of Positive Characteristic
Authors: Shah Firdous A.
Published: February 2015

We present a notion of frame multiresolution analysis on local fields of positive characteristic based on the theory of shift-invariant spaces. In contrast to the standard setting, the associated subspace V0 of L2(K) has a frame, a collection of translates of the scaling function φ of the form φ(·-u(k)):k∈N0, where N0 is the set of nonnegative integers. We investigate certain properties of multiresolution subspaces which provides the quantitative criteria for the construction of frame multiresolution analysis (FMRA) on local fields of positive characteristic. Finally, we provide a characterization of wavelet frames associated with FMRA on local field K of positive characteristic using the shift-invariant space theory.
Synthesis of N,N-Diethyldithiocarbamate Nitrile Ethyl and the Chelating Behaviors with Metal Ions
Authors: Zhang Zhonghui , Shi Lijun , Deng Wen , Jiang Dengbang , Lan Yaozhong
Published: February 2015

The N,N-diethyldithiocarbamate nitrile ethyl (NND) was the nonionic polar collector, and it can synthesize the NND in dimethyl sulfoxide solvent. This method can effectively reduce the reaction intensity and the coefficient of the synthetic risk. The purity of NND which we synthesized is 94.23%, and the yield is 91.06%. UV analysis shows that the characteristic absorption peak wavelength of the NND is 276 nm, and its absorbance is 0.901. Based on the interaction of NND + Mn+(Mn+= Fe3+, Cu2+, Zn2+, Pb2+) and the quantum chemical calculation analysis of the NND and ethyl xanthate, we can conclude that the flotation performance of NND should be better than that of ethyl xanthate.
Quantitative Proteomics and Lipidomics Analysis of Endoplasmic Reticulum of Macrophage Infected with Mycobacterium tuberculosis
Authors: Saquib Najmuddin Mohd , Jamwal Shilpa , Midha Mukul Kumar , Verma Hirdya Narain , Manivel Venkatasamy
Published: February 2015

Even though endoplasmic reticulum (ER) stress associated with mycobacterial infection has been well studied, the molecular basis of ER as a crucial organelle to determine the fate of Mtb is yet to be established. Here, we have studied the ability of Mtb to manipulate the ultrastructural architecture of macrophage ER and found that the ER-phenotypes associated with virulent (H37Rv) and avirulent (H37Ra) strains were different: a rough ER (RER) with the former against a smooth ER (SER) with the later. Further, the functional attributes of these changes were probed by MS-based quantitative proteomics (133 ER proteins) and lipidomics (8 phospholipids). Our omics approaches not only revealed the host pathogen cross-talk but also emphasized how precisely Mtb uses proteins and lipids in combination to give rise to characteristic ER-phenotypes. H37Ra-infected macrophages increased the cytosolic Ca2+ levels by attenuating the ATP2A2 protein and simultaneous induction of PC/PE expression to facilitate apoptosis. However, H37Rv inhibited apoptosis and further controlled the expression of EST-1 and AMRP proteins to disturb cholesterol homeostasis resulting in sustained infection. This approach offers the potential to decipher the specific roles of ER in understanding the cell biology of mycobacterial infection with special reference to the impact of host response.
Intra-Articular Giant Synovial Osteochondroma: Case Reports of the Ankle and Knee Joint
Authors: Fornaciari Paolo , Schai Pascal A. , Niehaus Richard , Exner Ulrich G.
Published: February 2015

Two cases of giant intra-articular osteochondromas (knee and ankle joint) are reported; pathologically they are rare representations of synovial chondromatosis. A 17-year-old man presented with a tumorous mass which had been localized in his left ankle for many years, increasing in volume during the last months. The lesion was removed by posteromedial ankle arthrotomy. The second case was observed in a 39-year-old woman with a slow-growing mass in her right knee joint. The lesion was removed from the Hoffa fat pad by open anteromedial arthrotomy.
Cellular Automata Based Modeling for Evaluating Different Bus Stop Designs in China
Authors: Ding Haoyang , Wang Wei , Luo Tianming , Yang Zhen , Li Ye , Li Zhibin
Published: February 2015

A cellular automaton model is proposed to simulate mixed traffic flow composed of motor vehicles and bicycles near bus stops. Three typical types of bus stops which are common in China are considered in the model, including two types of curbside bus stops and one type of bus bay stops. Passenger transport capacity of three types of bus stops, which is applied to evaluate the bus stop design, is calculated based on the corresponding traffic flow rate. According to the simulation results, the flow rates of both motor vehicles and bicycles exhibit phase transition from free flow to the saturation one at the critical point. The results also show that the larger the interaction between motor vehicle and bicycle flow is near curbside bus stops, the more the value of saturated flows drops. Curbside bus stops are more suitable when the conflicts between two flows are small and the inflow rate of motor vehicles is low. On the contrary, bus bay stops should be applied due to their ability to reduce traffic conflicts. Findings of this study can provide useful suggestions on bus stop selection considering different inflow rate of motor vehicles and bicycles simultaneously.
Determination of Matrine in Rat Plasma after Oral Administration of Novel Korean Herbal Medicine KIOM-MA128 and Application of PK
Authors: Back Hyun-moon , Song Byungjeong , Chae Jung-woo , Yun Hwi-yeol , Ma Jin Yeul , Kwon Kwang-il
Published: February 2015

KIOM-MA128 is a novel Korean herbal medicine with antiatopic, anti-inflammatory, and antiasthmatic effects. Matrine is thought to be a potential chemical marker of KIOM-MA128, but pharmacokinetic studies on KIOM-MA128 had not been performed. This study describes a simple and rapid method using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to determine the concentration of matrine in rats plasma after administration of KIOM-MA128. The isocratic mobile phase consisted of methanol and distilled water, and the flow rate was 0.15 mL/min. The accuracy and precision of the assay, as well as stability tests, were performed in accordance with FDA regulations for the validation of bioanalytical methods. The half-life andTmaxof matrine after administration of KIOM-MA128 were 4.29 ± 2.20 h and 1.8 ± 1.23 h, respectively.CmaxandAUCinfof matrine after administration of KIOM-MA128 at 4 g/kg and 8 g/kg were 595.10 ± 182.91 ng/mL, 5336.77 ± 1503.84 ng/mL·h and 850.46 ± 120 ng/mL, 9583.10 ± 888.92 ng/mL·h, respectively. The validated method was successfully applied to a pharmacokinetic study in rats after oral administration of KIOM-MA128.
Total Flavonoids fromClinopodium chinense(Benth.) O. Ktze Protect against Doxorubicin-Induced CardiotoxicityIn VitroandIn Vivo
Authors: Chen Rong Chang , Xu Xu Dong , Zhi Liu Xue , Sun Gui Bo , Zhu Yin Di , Dong Xi , Wang Jian , Zhang Hai Jing , Zhang Qiang , Sun Xiao Bo
Published: February 2015

Doxorubicin has cardiotoxic effects that limit its clinical benefit in cancer patients. This study aims to investigate the protective effects of the total flavonoids fromClinopodium chinense(Benth.) O. Ktze (TFCC) against doxorubicin- (DOX-) induced cardiotoxicity. Male rats were intraperitoneally injected with a single dose of DOX (3 mg/kg) every 2 days for three injections. Heart samples were collected 2 weeks after the last DOX dose and then analyzed. DOX delayed body and heart growth and caused cardiac tissue injury, oxidative stress, apoptotic damage, mitochondrial dysfunction, and Bcl-2 expression disturbance. Similar experiments in H9C2 cardiomyocytes showed that doxorubicin reduced cell viability, increased ROS generation and DNA fragmentation, disrupted mitochondrial membrane potential, and induced apoptotic cell death. However, TFCC pretreatment suppressed all of these adverse effects of doxorubicin. Signal transduction studies indicated that TFCC suppressed DOX-induced overexpression of p53 and phosphorylation of JNK, p38, and ERK. Studies with LY294002 (a PI3K/AKT inhibitor) demonstrated that the mechanism of TFCC-induced cardioprotection also involves activation of PI3K/AKT. These findings indicated the potential clinical application of TFCC in preventing DOX-induced cardiac oxidative stress.
Immediate Weight-Bearing after Ankle Fracture Fixation
Authors: Firoozabadi Reza , Harnden Emily , Krieg James C.
Published: February 2015

We believe that a certain subset of surgical ankle fracture patients can be made weight-bearing as tolerated immediately following surgery. Immediate weight-bearing as tolerated (IWBAT) allows patients to return to ambulation and activities of daily living faster and may facilitate rehabilitation. A prospectively gathered orthopaedic trauma database at a Level 1 trauma center was reviewed retrospectively to identify patients who had ORIF after unstable ankle injuries treated by the senior author. Patients were excluded if they were not IWBAT based on specific criteria or if they did meet followup requirement. Only 1/26 patients was noted to have loss of fixation. This was found at the 6-week followup and was attributed to a missed syndesmotic injury. At 2-week followup, 2 patients had peri-incisional erythema that resolved with a short course of oral antibiotics. At 6-week followup, 20 patients were wearing normal shoes and 6 patients continued to wear the CAM Boot for comfort. To conclude, IWBAT in a certain subset of patients with stable osteosynthesis following an ankle fracture could potentially be a safe alternative to a period of protected weight-bearing.
A Lipid Mediator Hepoxilin A3 Is a Natural Inducer of Neutrophil Extracellular Traps in Human Neutrophils
Authors: Douda David N. , Grasemann Hartmut , Pace-Asciak Cecil , Palaniyar Nades
Published: February 2015

Pulmonary exacerbations in cystic fibrosis airways are accompanied by inflammation, neutrophilia, and mucous thickening. Cystic fibrosis sputum contains a large amount of uncleared DNA contributed by neutrophil extracellular trap (NET) formation from neutrophils. The exact mechanisms of the induction of NETosis in cystic fibrosis airways remain unclear, especially in uninfected lungs of patients with early cystic fibrosis lung disease. Here we show that Hepoxilin A3, a proinflammatory eicosanoid, and the synthetic analog of Hepoxilin B3, PBT‐3, directly induce NETosis in human neutrophils. Furthermore, we show that Hepoxilin A3‐mediated NETosis is NADPH‐oxidase‐dependent at lower doses of Hepoxilin A3, while it is NADPH‐oxidase‐independent at higher doses. Together, these results demonstrate that Hepoxilin A3 is a previously unrecognized inducer of NETosis in cystic fibrosis lungs and may represent a new therapeutic target for treating cystic fibrosis and other inflammatory lung diseases.
Does Probability Guided Hysteroscopy Reduce Costs in Women Investigated for Postmenopausal Bleeding?
Authors: Breijer M. C. , van Hanegem N. , Visser N. C. M. , Verheijen R. H. M. , Mol B. W. J. , Pijnenborg J. M. A. , Opmeer B. C. , Timmermans A.
Published: February 2015

Objective. To evaluate whether a model to predict a failed endometrial biopsy in women with postmenopausal bleeding (PMB) and a thickened endometrium can reduce costs without compromising diagnostic accuracy. Design, Setting, and Population. Model based cost‐minimization analysis. Methods. A decision analytic model was designed to compare two diagnostic strategies for women with PMB: (I) attempting office endometrial biopsy and performing outpatient hysteroscopy after failed biopsy and (II) predicted probability of a failed endometrial biopsy based on patient characteristics to guide the decision for endometrial biopsy or immediate hysteroscopy. Robustness of assumptions regarding costs was evaluated in sensitivity analyses. Main Outcome Measures. Costs for the different strategies. Results. At different cut‐offs for the predicted probability of failure of an endometrial biopsy, strategy I was generally less expensive than strategy II. The costs for strategy I were always € 460; the costs for strategy II varied between € 457 and € 475. At a 65% cut‐off, a possible saving of € 3 per woman could be achieved. Conclusions. Individualizing the decision to perform an endometrial biopsy or immediate hysteroscopy in women presenting with postmenopausal bleeding based on patient characteristics does not increase the efficiency of the diagnostic work‐up.
Physicochemical Properties and Antioxidant Activity of Chitin and Chitosan Prepared from Pacific White Shrimp Waste
Authors: Si Trung Trang , Bao Huynh Nguyen Duy
Published: February 2015

Chitin and chitosan, valuable marine biopolymers, recovered from shrimp waste, are an abundant by-product of the shrimp processing industry in Vietnam, at an estimated 200000 metric tons per year. The obtained chitin and chitosan are characterized by their purity and functional properties. The polymers show good quality with low residual ash and protein content (<1%). The antioxidant potency of chitosan is evaluated by several different in vitro systems, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, total reducing power, and inhibition of lipid peroxidation. The DPPH free radical scavenging, total reducing power, and lipid peroxidation inhibition activities of chitosan at varying concentration (0.125 to 1.0 mg/mL) range from 3.7 to 16.8%, 0.05 to 0.15, and 1.7 to 15.1%, respectively. This study demonstrates that chitin and chitosan, of good quality and having characteristics compatible with a broad range of applications, can be prepared from white shrimp waste.
Cutaneous Ulcer as Leading Symptom of Systemic Cytomegalovirus Infection
Authors: Guo Richard F. , Gebreab Frew H. , Tang Emily Hsiang-Ho , Piao Zhe , Lee Steve S. , Perez Mario L.
Published: February 2015

Cytomegalovirus (CMV) infection rarely manifests with skin ulcerations. We describe a case report of a 64-year-old woman with chronic immunosuppression for treatment of mixed connective tissue disease, presenting with new onset leg ulcerations after a recent change in immunosuppressive regimen. She subsequently developed fulminant hepatitis, encephalopathy, and pancytopenia and was found to have severe systemic CMV viremia. Skin ulcer biopsy was positive by immunohistochemical staining for CMV infected endothelial cells. Both systemic disease and skin ulcer rapidly improved after stopping immunosuppression and administering intravenous ganciclovir. New onset skin ulcers in an immunosuppressed individual, especially with recent changes in immunosuppressive regimen, should raise the suspicion of reactivation of CMV.
Antihypertensive Effect of Radix Paeoniae Alba in Spontaneously Hypertensive Rats and Excessive Alcohol Intake and High Fat Diet Induced Hypertensive Rats
Authors: Su-Hong Chen , Qi Chen , Bo Li , Jian-Li Gao , Jie Su , Gui-Yuan Lv
Published: February 2015

Radix Paeoniae Alba (Baishao, RPA) has long been used in traditional Chinese medicine formulation to treat hypertension by repression the hyperfunction of liver. However, whether the RPA itself has the antihypertensive effect or not is seldom studied. This study was to evaluate the protective effect of RPA on hypertensive rats. Alcohol in conjunction with a high fat diet- (ACHFD-) induced hypertensive rats and spontaneously hypertensive rats (SHR) was constantly received either RPA extract (25 or 75 mg/kg) or captopril (15 mg/kg) all along the experiments. As a result, RPA extract (75 mg/kg) could significantly reduce systolic blood pressure of both ACHFD-induced hypertensive rats and SHR after 9-week or 4-week treatment. In ACHFD-induced hypertensive rats, the blood pressure was significantly increased and the lipid profiles in serum including triglyceride, total cholesterol, LDL-cholesterol, and HDL-cholesterol were significantly deteriorated. Also, hepatic damage was manifested by a significant increase in alanine transaminase (ALT) and aspartate transaminase (AST) in serum. The RPA extract significantly reversed these parameters, which revealed that it could alleviate the liver damage of rats. In SHR, our result suggested that the antihypertensive active of RPA extract may be related to its effect on regulating serum nitric oxide (NO) and endothelin (ET) levels.
Maximum Likelihood Sequence Detection Receivers for Nonlinear Optical Channels
Authors: Maggio Gabriel N. , Hueda Mario R. , Agazzi Oscar E.
Published: February 2015

The space-time whitened matched filter (ST-WMF) maximum likelihood sequence detection (MLSD) architecture has been recently proposed (Maggio et al., 2014). Its objective is reducing implementation complexity in transmissions over nonlinear dispersive channels. The ST-WMF-MLSD receiver (i) drastically reduces the number of states of the Viterbi decoder (VD) and (ii) offers a smooth trade-off between performance and complexity. In this work the ST-WMF-MLSD receiver is investigated in detail. We show that thespace compressionof the nonlinear channel is an instrumental property of the ST-WMF-MLSD which results in a major reduction of the implementation complexity in intensity modulation and direct detection (IM/DD) fiber optic systems. Moreover, we assess the performance of ST-WMF-MLSD in IM/DD optical systems with chromatic dispersion (CD) and polarization mode dispersion (PMD). Numerical results for a 10 Gb/s, 700 km, and IM/DD fiber-optic link with 50 ps differential group delay (DGD) show that the number of states of the VD in ST-WMF-MLSD can be reduced ~4 times compared to an oversampled MLSD. Finally, we analyze the impact of the imperfect channel estimation on the performance of the ST-WMF-MLSD. Our results show that the performance degradation caused by channel estimation inaccuracies is low and similar to that achieved by existing MLSD schemes (~0.2 dB).
Finite Element Assembly Using an Embedded Domain Specific Language
Authors: Janssens Bart , Bányai Támas , Limam Karim , Bosschaerts Walter
Published: February 2015

In finite element methods, numerical simulation of the problem requires the generation of a linear system based on an integral form of a problem. Using C++ meta-programming techniques, a method is developed that allows writing code that stays close to the mathematical formulation. We explain the specifics of our method, which relies on the Boost.Proto framework to simplify the evaluation of our language. Some practical examples are elaborated, together with an analysis of the performance. The abstraction overhead is quantified using benchmarks.
Integrated Production and Distribution Scheduling Problems Related to Fixed Delivery Departure Dates and Weights of Late Orders
Authors: Li Shanlin , Li Maoqin
Published: February 2015

We consider an integrated production and distribution scheduling problem faced by a typical make‐to‐order manufacturer which relies on a third‐party logistics (3PL) provider for finished product delivery to customers. In the beginning of a planning horizon, the manufacturer has received a set of orders to be processed on a single production line. Completed orders are delivered to customers by a finite number of vehicles provided by the 3PL company which follows a fixed daily or weekly shipping schedule such that the vehicles have fixed departure dates which are not part of the decisions. The problem is to find a feasible schedule that minimizes one of the following objective functions when processing times and weights are oppositely ordered: (1) the total weight of late orders and (2) the number of vehicles used subject to the condition that the total weight of late orders is minimum. We show that both problems are solvable in polynomial time.
Phytochemical Analysis and Modulation of Antibiotic Activity byLuehea paniculataMart. & Zucc. (Malvaceae) in Multiresistant Clinical Isolates ofCandidaSpp.
Authors: Calixto Júnior João T. , Morais Selene M. , Martins Clécio G. , Vieira Larissa G. , Morais-Braga Maria Flaviana B. , Carneiro Joara N. P. , Machado Antonio J. P. , Menezes Irwin R. A. , Tintino Saulo R. , Coutinho Henrique D. M.
Published: February 2015

The high incidence of fungal infections has led to the continuous search for new drugs. Extracts ofLuehea paniculata, a tree of multiple medicinal uses, were evaluated for anti-Candidaactivity, as well as its modulator potential of the Fluconazole antibiotic. Chemical prospecting of ethanol extracts of leaf and bark was carried out, the quantification of total phenols and flavonoids, characterized by the HPLC-DAD technique. The rosmarinic acid and the vitexin flavonoid were observed as major constituents in ELELP and ESWELP, respectively. Antioxidant activity was also evaluated by the method of scavenging the free radical DPPH, and quercetin was used as standard, obtaining IC50values: 0.341 (mg/mL) for ELELP and 0.235 (mg/mL) for ESWELP. The microdilution assay was performed for antifungal activity against strains ofCandida albicans, C. krusei, andC. tropicalisand showed minimum inhibitory concentrations values ≥1024 μg/mL. In the modulator action of extracts on Fluconazole against multiresistant clinical isolates ofCandida(subinhibitory concentration minimum of 128 μg/mL), a significant synergism was observed, indicating that the extracts potentiated the antifungal effect againstC. tropicalis, where antioxidant flavonoids could be responsible. This is the first report about modifying activity of the antibiotic action of a species of the genusLuehea.
Adenosine A2A Receptors Mediate Anti-Inflammatory Effects of Electroacupuncture on Synovitis in Mice with Collagen-Induced Arthritis
Authors: Li Qi-hui , Xie Wen-xia , Li Xiao-pei , Huang Ka-te , Du Zhong-heng , Cong Wen-jie , Zhou Long-hua , Ye Tian-shen , Chen Jiang-Fan
Published: February 2015

To study the role of adenosine A2A receptor (A2AR) in mediating the anti-inflammatory effect of electroacupuncture (EA) on synovitis in collagen-induced arthritis (CIA), C57BL/6 mice were divided into five treatment groups:Sham-control, CIA-control, CIA-EA, CIA-SCH58261(A2ARantagonist), andCIA-EA-SCH58261. All mice except those in theSham-controlgroup were immunized with collagen II for arthritis induction. EA treatment was administered using the stomach 36 and spleen 6 points, and stimulated with a continuous rectangular wave for 30 min daily. EA treatment and SCH58261 were administered daily from days 35 to 49 (n=10). After treatment, X-ray radiography of joint bone morphology was established at day 60 and mouse blood was collected for ELISA determination of tumor necrosis factor alpha (TNF-α) levels. Mice were sacrificed and processed for histological examination of pathological changes of joint tissue, including hematoxylin-eosin staining and immunohistochemistry ofA2ARexpression. EA treatment resulted in significantly reduced pathological scores, TNF-αconcentrations, and bone damage X-ray scores. Importantly, the anti-inflammatory and tissue-protective effect of EA treatment was reversed by coadministration of SCH58261. Thus, EA treatment exerts an anti-inflammatory effect resulting in significant protection of cartilage by activation ofA2ARin the synovial tissue of CIA.
Recent Advances in the Diagnosis and Treatment of Niemann-Pick Disease Type C in Children: A Guide to Early Diagnosis for the General Pediatrician
Authors: Alobaidy Hanna
Published: February 2015

Niemann-Pick disease (NP-C) is a lysosomal storage disease in which impaired intracellular lipid transport leads to accumulation of cholesterol and glycosphingolipids in various neurovisceral tissues. It is an autosomal recessive disorder, caused by mutations in theNPC1orNPC2genes. The clinical spectrum is grouped by the age of onset and onset of neurological manifestation: pre/perinatal; early infantile; late infantile; and juvenile periods. The NP-C Suspicion Index (SI) screening tool was developed to identify suspected patients with this disease. It is especially good at recognizing the disease in patients older than four years of age. Biochemical tests involving genetic markers and Filipin staining of skin fibroblast are being employed to assist diagnosis. Therapy is mostly supportive and since 2009, the first specific therapy approved for use was Miglustat (Zavesca) aimed at stabilizing the rate of progression of neurological manifestation. The prognosis correlates with age at onset of neurological signs; patients with early onset form progress faster. The NP-C disease has heterogeneous neurovisceral manifestations. A SI is a screening tool that helps in diagnostic process. Filipin staining test is a specific biomarker diagnostic test. Miglustat is the first disease-specific therapy.
Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European BatLyssavirusType 1
Authors: Picard-Meyer Evelyne , Peytavin de Garam Carine , Schereffer Jean Luc , Marchal Clotilde , Robardet Emmanuelle , Cliquet Florence
Published: February 2015

This study evaluates the performance of five two-step SYBR Green RT-qPCR kits and five one-step SYBR Green qRT-PCR kits using real-time PCR assays. Two real-time thermocyclers showing different throughput capacities were used. The analysed performance evaluation criteria included the generation of standard curve, reaction efficiency, analytical sensitivity, intra- and interassay repeatability as well as the costs and the practicability of kits, and thermocycling times. We found that the optimised one-step PCR assays had a higher detection sensitivity than the optimised two-step assays regardless of the machine used, while no difference was detected in reaction efficiency,R2values, and intra- and interreproducibility between the two methods. The limit of detection at the 95% confidence level varied between 15 to 981 copies/µL and 41 to 171 for one-step kits and two-step kits, respectively. Of the ten kits tested, the most efficient kit was the Quantitect SYBR Green qRT-PCR with a limit of detection at 95% of confidence of 20 and 22 copies/µL on the thermocyclers Rotor gene Q MDx and MX3005P, respectively. The study demonstrated the pivotal influence of the thermocycler on PCR performance for the detection of rabies RNA, as well as that of the master mixes.
Intercalary Reconstruction after Wide Resection of Malignant Bone Tumors of the Lower Extremity Using a Composite Graft with a Devitalized Autograft and a Vascularized Fibula
Authors: Ogura Koichi , Miyamoto Shimpei , Sakuraba Minoru , Fujiwara Tomohiro , Chuman Hirokazu , Kawai Akira
Published: February 2015

Introduction. Although several intercalary reconstructions after resection of a lower extremity malignant bone tumor are reported, there are no optimal methods which can provide a long-term reconstruction with fewest complications. We present the outcome of reconstruction using a devitalized autograft and a vascularized fibula graft composite.Materials and Methods.We conducted a retrospective review of 11 patients (7 males, 4 females; median age 27 years) undergoing reconstruction using a devitalized autograft (pasteurization (n=6), deep freezing (n=5)) and a vascularized fibula graft composite for lower extremity malignant bone tumors (femur (n=10), tibia (n=1)).Results. The mean period required for callus formation and bone union was 4.4 months and 9.9 months, respectively. Four postoperative complications occurred in 3 patients: 2 infections (1 pasteurized autograft, 1 frozen autograft) and 1 fracture and 1 implant failure (both in pasteurized autografts). Graft removal was required in 2 patients with infections. The mean MSTS score was 81% at last follow-up.Conclusions.Although some complications were noted in early cases involving a pasteurized autograft, our novel method involving a combination of a frozen autograft with a vascularized fibula graft and rigid fixation with a locking plate may offer better outcomes than previously reported allografts or devitalized autografts.
Fast HPLC-MS/MS Method for Determining Penicillin Antibiotics in Infant Formulas Using Molecularly Imprinted Solid-Phase Extraction
Authors: Díaz-Bao Mónica , Barreiro Rocío , Miranda José Manuel , Cepeda Alberto , Regal Patricia
Published: February 2015

The dairy cattle may suffer from different infections relatively often, but the inflammation of the mammary gland is very important to the farmer. These infections are frequently treated with penicillin antimicrobial drugs. However, their use may result in the presence of residues in animal products, such as milk powder and/or infant formulas, and it represents a potential risk for consumers. To monitor this, the EU has defined safe maximum residue limits (MRLs) through Commission Regulation (EU) number 37/2010. Although LC-MS is a trustful option for confirmation and quantification of antibiotics, the analysis of real samples with complex matrices frequently implies previous clean-up steps. In this work, precipitation polymerization has been used and different molecularly imprinted polymer (MIP) sorbents were tested and optimized for the fast and simultaneous solid-phase extraction (MISPE) of eight common penicillins (ampicillin, amoxicillin, oxacillin, penicillin G, penicillin V, cloxacillin, dicloxacillin, and nafcillin). The extracts were analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) and the applicability of these polymers as sorbents for the extraction of penicillins at MRL levels in milk powder (infant formulas) was proved. The limits of detection and quantification were below the legal tolerances, except for LOQ for oxacillin and cloxacillin.
Suppressing TGFβ Signaling in Regenerating Epithelia in an Inflammatory Microenvironment Is Sufficient to Cause Invasive Intestinal Cancer
Authors: Oshima Hiroko , Nakayama Mizuho , Han Tae-Su , Naoi Kuniko , Ju Xiaoli , Maeda Yusuke , Robine Sylvie , Tsuchiya Kiichiro , Sato Toshiro , Sato Hiroshi , Taketo Makoto Mark , Oshima Masanobu
Published: February 2015

Genetic alterations in the TGFβ signaling pathway in combination with oncogenic alterations lead to cancer development in the intestines. However, the mechanisms of TGFβ signaling suppression in malignant progression of intestinal tumors have not yet been fully understood. We have examined ApcΔ716 Tgfbr2ΔIEC compound mutant mice that carry mutations in Apc and Tgfbr2 genes in the intestinal epithelial cells. We found inflammatory microenvironment only in the invasive intestinal adenocarcinomas but not in noninvasive benign polyps of the same mice. We thus treated simple Tgfbr2ΔIEC mice with dextran sodium sulfate (DSS) that causes ulcerative colitis. Importantly, these Tgfbr2ΔIEC mice developed invasive colon cancer associated with chronic inflammation. We also found that TGFβ signaling is suppressed in human colitis–associated colon cancer cells. In the mouse invasive tumors, macrophages infiltrated and expressed MT1-MMP, causing MMP2 activation. These results suggest that inflammatory microenvironment contributes to submucosal invasion of TGFβ signaling–repressed epithelial cells through activation of MMP2. We further found that regeneration was impaired in Tgfbr2ΔIEC mice for intestinal mucosa damaged by DSS treatment or X-ray irradiation, resulting in the expansion of undifferentiated epithelial cell population. Moreover, organoids of intestinal epithelial cells cultured from irradiated Tgfbr2ΔIEC mice formed “long crypts” in Matrigel, suggesting acquisition of an invasive phenotype into the extracellular matrix. These results, taken together, indicate that a simple genetic alteration in the TGFβ signaling pathway in the inflamed and regenerating intestinal mucosa can cause invasive intestinal tumors. Such a mechanism may play a role in the colon carcinogenesis associated with inflammatory bowel disease in humans. Cancer Res; 75(4); 766–76. ©2015 AACR.
A Rolling Circle Amplification Screen for Polyomaviruses Other than BKPyV in Renal Transplant Recipients Confirms High Prevalence of Urinary JCPyV Shedding
Authors: Kluba Jeanette , Linnenweber-Held Silvia , Heim Albert , Ang Angella M. , Raggub Lubna , Broecker Verena , Becker Jan U. , Schulz Thomas F. , Schwarz Anke , Ganzenmueller Tina
Published: February 2015

Objectives: Multiple novel human polyomaviruses (HPyVs) have been discovered in the last few years. These or other, unknown, nephrotropic HPyVs may potentially be shed in urine. Methods: To search for known and unknown HPyVs we investigated BKPyV-negative urine samples from 105 renal transplant recipients (RTR) by rolling circle amplification (RCA) analysis and quantitative JCPyV PCR. Clinical data was analysed to identify risk factors for urinary polyomavirus shedding. Results: In 10% (11/105) of the urine samples RCA with subsequent sequencing revealed JCPyV, but no other HPyV sequences. Using quantitative JCPyV PCR, 24% (25/105) of the samples tested positive. Overall sensitivities of RCA of 44% (11/25) in detecting JCPyV in JCPyV DNA-positive urine and 67% (10/15) for samples with JCPyV loads >10,000 copies/ml can be assumed. Despite frequent detectable urinary shedding of JCPyV in our cohort, this could not be correlated with clinical risk factors. Conclusion: Routine urinary JCPyV monitoring in BKPyV-negative RTR without suspected polyomavirus-associated nephropathy might be of limited diagnostic value. As RCA works in a sequence-independent manner, detection of novel and known polyomaviruses shed in sufficient quantities is feasible. High-level shedding of HPyVs other than BKPyV or JCPyV in the urine of RTR is unlikely to occur.
Meconium Aspiration Syndrome: Possible Pathophysiological Mechanisms and Future Potential Therapies
Authors: Haakonsen Lindenskov Paal Helge , Castellheim Albert , Saugstad Ola Didrik , Mollnes Tom Eirik
Published: February 2015

Does meconium cause meconium aspiration syndrome (MAS) or is meconium discharge only a marker of fetal hypoxia? This dispute has lasted for centuries, but since the 1960s, detrimental effects of meconium itself on the lungs have been demonstrated in animal experiments. In clinical MAS, persistent pulmonary hypertension of the newborn is the leading cause of death in MAS. Regarding the complex chemical composition of meconium, it is difficult to identify a single agent responsible for the pathophysiology. However, considering that meconium is stored in the intestines, partly unexposed to the immune system, aspirated meconium could be recognized as ‘danger', representing damaged self. The common denominator in the pathophysiology could therefore be activation of innate immunity. Thus, a bulk of evidence implies that meconium is a potent activator of inflammatory mediators, including cytokines, complement, prostaglandins and reactive oxygen species. We hypothesize that the two main recognition systems of innate immunity, the Toll-like receptors and the complement system, recognize meconium as ‘danger', which leads not only to lung dysfunction but also to a systemic inflammatory response. This might have therapeutic implications in the future.
Integrative Cardiac Reserve
Authors: Xue Jingui , Zhao Feiran , Wang Yuqi , Gu Jialin , Gao Junjie , Wang Xiaolong , Zhou Hua
Published: February 2015

Recent progress suggests that integrative cardiac reserve is important in coronary artery disease, chronic heart failure, cardiomyopathy, and other cardiovascular diseases. Integrative cardiac reserve includes coronary flow reserve, myocardial reserve, energy reserve, and heart rate reserve. In the case of stress reaction, coronary flow reserve can provide excessive blood and energy supply and increase the heart rate. Thus, the dysfunctional myocardium can recover its contractile characteristics. In this study, we review the current knowledge on integrative cardiac reserve in order to provide an additional cure and strategy for the treatment of heart diseases.
Generating Conditionally Immortalised Podocyte Cell Lines from Wild-Type Mice
Authors: Keir Lindsay S. , Firth Rachel , May Carl , Ni Lan , Welsh Gavin I. , Saleem Moin A.
Published: February 2015

Background: Understanding podocyte biology is key to deciphering the pathogenesis of numerous glomerular diseases. However, cultivation of primary podocytes results in de-differentiation with loss of specialised architecture. Human conditionally immortalised podocytes partly overcome this problem, utilising a temperature-sensitive transgene. Conditionally immortalised murine podocytes exist, but are derived from the Immortomouse. Methods: Using retroviral temperature-sensitive SV40 transfection, we created a conditionally immortalised podocyte cell line from wild-type mice. Results: These cells develop characteristic mature podocyte morphology and robustly express slit diaphragm proteins. Functionally, these cells demonstrate comparable responses in motility and glucose uptake to human conditionally immortalised podocytes. Conclusion: Podocyte-specific transgenic mice are extensively used to study glomerular disease and this technique could be used to make podocyte cell lines from any mouse, allowing study at the cellular level. This will help characterise these disease models and add to the laboratory resources used to study podocytopathies and glomerular disease.
Effects of Food-Derived Collagen Peptides on the Expression of Keratin and Keratin-Associated Protein Genes in the Mouse Skin
Authors: Le Vu Phuong , Takatori Ryo , Iwamoto Taku , Akagi Yutaka , Satsu Hideo , Totsuka Mamoru , Chida Kazuhiro , Sato Kenji , Shimizu Makoto
Published: February 2015

Oral ingestion of collagen peptides (CP) has long been suggested to exert beneficial effects on the skin, but the molecular events induced by CP on the skin remain unclear. Here, we investigated the effects of oral CP administration on gene expression in hairless mouse skin and of prolyl-hydroxyproline (Pro-Hyp), a collagen-derived dipeptide, on gene expression in a coculture of mouse skin keratinocytes and fibroblasts. Using microarray analysis, we found that oral administration of CP to hairless mice for 6 weeks induced increased expression of Krtap and Krt genes in the skin. Annotation analysis using DAVID revealed that a group of the up-regulated genes, Gprc5d, Sprr2a1, Krt27 and Krtap16-7, is associated with the development of the epidermis and the hair cycle. In addition, the presence of Pro-Hyp (200 μM) induced an increase in the expression of Krtap16-7, Krtap15, Krtap14 and Krtap8-2 in keratinocytes in coculture, partially resembling the in vivo result. The Pro-Hyp-induced up-regulation of these genes was not observed when keratinocytes were cultured without fibroblasts, suggesting that the presence of fibroblasts is essential for the effects of Pro-Hyp. Our study presents new insights into the effects of CP on the skin, which might link to the hair cycle.
Pulmonary Rehabilitation and Noninvasive Ventilation in Patients with Hypercapnic Interstitial Lung Disease
Authors: Dreher Michael , Ekkernkamp Emelie , Schmoor Claudia , Schoenheit-Kenn Ursula , Winterkamp Sandra , Kenn Klaus
Published: February 2015

Background: Pulmonary rehabilitation (PR) has a positive impact on functional status and quality of life in patients with interstitial lung disease (ILD). Objectives: This study investigated the effects of PR in hypercapnic ILD patients receiving nighttime noninvasive positive pressure ventilation (NPPV). Methods: Consecutive ILD patients referred to a specialized inpatient PR center were included. All participated in a PR program. Those with hypercapnia received NPPV (NPPV group; n = 29); the remaining patients served as comparison group (n = 319). Results: PR improved the 6-min walk distance by 64.4 ± 67.1 m versus baseline (p < 0.0001) in NPPV patients and by 43.2 ± 55.1 m (p < 0.0001) in the comparison group (difference 21.1 m, 95% confidence interval 0.5-41.8; p = 0.045). There was no change in total lung capacity during PR in NPPV recipients or the comparison group. Forced vital capacity significantly increased from baseline in the comparison, but not the NPPV group. NPPV recipients were significantly more likely than the comparison group to have improved dyspnea during PR (p = 0.049). There was no improvement in the 36-item Short Form (SF-36) physical component score in the NPPV group after PR, but there was in the comparison group. PR improved the SF-36 mental component score versus baseline in both groups. Conclusion: An individually tailored PR plus nighttime NPPV appears feasible in hypercapnic ILD patients and significantly improves exercise capacity and quality of life.
Semicircular Lipoatrophy: An Electrostatic Hypothesis
Authors: Linares-García Valdecasas Rafael , Cuerda-Galindo Esther , Ramiro Bargueño Julio , Naranjo Garcia Pablo , Vogelfrang-Garncarz Daniel , Palomar-Gallego M. Angustias
Published: February 2015

Background: Semicircular lipoatrophy (SL) is a benign pathology characterized by subcutaneous adipose tissue atrophy. Objective: To determine causes of SL. Methods: We performed research on 42 cases of SL who had developed symptoms of lipoatrophy with a technical study of the building and clinical study of the affected patients. Results: Electromagnetic field and electrostatic charge measurements were performed. No signals different from typical radio, TV and other wireless communication devices were recorded. Electrostatic charges were recorded in floors, environments and on different furniture; they were positive, and all charges disappeared on contact with the worker's skin. Data about the workplace were collected, and clinical examinations were performed, including blood and biochemistry tests and 18 ultrasound skin tests. Conclusions: As no abnormal electromagnetic measurements were found and electrostatic changes and occupational behaviors showed some relevant data and as after changes in order to control the electrostatic environment all of them improved, we can conclude that electrostatic charges but not electromagnetic ones are the main cause of SL.
Transcriptional Analysis of Host Responses to Marek's Disease Virus Infection in Chicken Thymus
Authors: Hu Xuming , Qin Aijian , Xu Wencai , Wu Genghua , Li Dan , Qian Kun , Shao Hongxia , Ye Jianqiang
Published: February 2015

Marek's disease virus (MDV) is a cell-associated alpha-herpesvirus that causes T-cell lymphomas and nervous disorders in chickens. Different from other lymphoid organs, the thymus is the site of T-cell maturation and differentiation. However, the transcriptional response to MDV infection in the chicken thymus is still not known. In this study, we performed genome-wide expression analysis in thymus tissues of RB1B-infected chickens at different time points to investigate the molecular mechanisms of MDV pathogenesis. The number of differentially expressed genes with 2-fold or higher changes (>2) are as follows: 1,250 genes (7 dpi), 834 genes (14 dpi), 1,958 genes (21 dpi), and 2,306 genes (28 dpi). Gene ontology enrichment analysis revealed that the upregulated genes were involved in immune and inflammatory response at 7 dpi; angiogenesis, cytoskeleton organization, cell adhesion, and signal transduction showed different expressions at 21 and 28 dpi. The expression pattern of 18 randomly selected genes was confirmed by real-time RT-PCR. Several differently expressed host genes associated with tumor development are discussed. We identified the global host-gene expression pattern in the thymus of chickens that responded to MDV infection. The present data may provide groundwork for future investigation in the biology and pathogenesis of MDV.
Murine Gammaherpesvirus (MHV-68) Transforms Cultured Cells in vitro
Authors: Mrázová Veronika , Betáková Tatiana , Kúdelová Marcela , Šupolíková Miroslava , Lachová Veronika , Lapuníková Barbora , Golais František
Published: February 2015

Human dermal fibroblasts and mouse NIH/3T3 cells acquired the transformed phenotype (‘criss-cross' pattern of growth) after infection with ultraviolet-irradiated murine gammaherpesvirus (MuHV-4 strain 68; MHV-68). These cells with changed phenotype could be serially cultured for 5-6 passages (35-40 days), and then they entered into crisis and most of them died. In a small number of cultures, however, foci of newly transformed cells appeared from which two stable cell lines were derived. After 6-9 cell culture passages of the MHV-68 transformed cell lines, MHV-68 DNA and virus antigen could be detected by PCR and immunofluorescence assay along with the disappearance of actin bundles, indicating that both transformed cell lines might be oncogenic.
In vivo Reflectance Confocal Microscopy Characterization of Field-Directed 5-Fluorouracil 0.5%/Salicylic Acid 10% in Actinic Keratosis
Authors: Ulrich Martina , Alarcon Ivette , Malvehy Josep , Puig Susana
Published: February 2015

Actinic keratosis (AK), a frequently diagnosed cutaneous neoplasm in individuals with chronic sun exposure or fair skin, is a risk factor for squamous cell carcinoma. AK presents as clinically visible lesions and/or as subclinical lesions where an entire field of area (field cancerization) contains lesions of various grades. The diagnosis and surveillance of subclinical AK is challenging. We report a new AK management approach, including subclinical AK, with noninvasive in vivo reflectance confocal microscopy (RCM) monitoring of field-directed topical 5-fluorouracil 0.5%/salicylic acid 10.0% (5-FU/SA; currently approved for single lesions). In this case series, eight patients with primarily recurrent, multiple AKs received ≤6 weeks of field-directed 5-FU/SA; complete clearance of clinical/subclinical AKs on various body areas was shown in most patients using RCM. RCM facilitated the detection/characterization of subclinical AKs in the setting of field cancerization. Topical field-directed 5-FU/SA monitored with RCM is a promising management approach for subclinical AKs.