bluuebunny/crossref_metadata_2025 huggingface dataset:
Streptozotocin‐Induced Maternal Hyperglycemia Increases the Expression of Antioxidant Enzymes and Mast Cell Number in Offspring Rat Ventral Prostate
Authors: Camargo Ana C. L. , Dos Santos Sérgio A. A. , Rinaldi Jaqueline C. , Constantino Flavia B. , Colombelli Ketlin T. , Scarano Wellerson R. , Felisbino Sérgio L. , Justulin Luis A.
Published: November 2016

Gestational diabetes mellitus (GDM) has increased in recent years. Although the cellular and molecular mechanisms involved in GDM‐increased risk factors to offspring remained poorly understood, some studies suggested an association between an increase in oxidative stress induced by maternal hyperglycemia and complications for both mothers and newborns. Here, we investigated the impact of maternal hyperglycemia followed by maternal insulin replacement during lactation on the expression of antioxidant enzymes and mast cell number in offspring ventral prostate (VP) at puberty. Pregnant rats were divided into three groups: control (CT); streptozotocin‐induced maternal hyperglycemia (MH); and MH plus maternal insulin replacement during lactation (MHI). Male offspring were euthanized at postnatal day (PND) 60 and the VP was removed and processed for histology and Western blotting analyses. Maternal hyperglycemia delayed prostate maturation, and increased mast cell number catalase (CAT), superoxide dismutase (SOD), glutatione‐s‐transferase (GST‐pi), and cyclooxygenase‐2 (Cox‐2) expression in the offspring of hyperglycemic dams. Maternal insulin replacement restored VP structure, mast cell number and antioxidant protein expression, except for Cox‐2, which remained higher in the MHI group. Thus, an increase in oxidative stress induced by intrauterine hyperglycemia impacts prostate development and maturation, which persists until puberty. The overall improvement of maternal metabolism after insulin administration contributes to the restoration of prostate antioxidant enzymes and secretory function. Taken together, our results highlighted that imbalanced physiological maternal‐fetal interaction contributes to the impairment of reproductive performance of the offspring from diabetic mothers. Anat Rec, 300:291–299, 2017. © 2016 Wiley Periodicals, Inc.
Ultraviolet B Irradiation Causes Stimulator of Interferon Genes–Dependent Production of Protective Type I Interferon in Mouse Skin by Recruited Inflammatory Monocytes
Authors: Sontheimer Clayton , Liggitt Denny , Elkon Keith B.
Published: November 2016

Objective

Photosensitivity is common in patients with systemic lupus erythematosus, although the mechanisms linking ultraviolet (UV) light to flares are not well understood. We undertook this study to determine whether repetitive UVB exposure could induce type I interferon (IFN) production in normal mouse skin, and to investigate the roles of inflammatory monocytes and plasmacytoid dendritic cells (PDCs) in type I IFN production and development of UVB irradiation–induced inflammation.

Methods

Mice were irradiated with UVB at 100 mJ/cm2 for 5 days, and cutaneous manifestations were examined by messenger RNA expression of inflammatory and type I IFN response genes, histology, and flow cytometry. Inflammatory monocyte and PDC depletion experiments were performed in CCR2–diphtheria toxin receptor (DTR)–transgenic mice and blood dendritic cell antigen 2–DTR–transgenic mice. The roles of type I IFN and of the adaptor protein stimulator of IFN genes (STING) in UVB irradiation–induced inflammation were investigated using IFN‐α/β/ω receptor (IFNAR)–knockout mice and STING‐knockout mice.

Results

Repeated UVB irradiation stimulated an inflammatory cell infiltrate and induction of type I IFN and proinflammatory cytokines. Interestingly, the type I IFN response was independent of PDCs but dependent on inflammatory monocytes, which were recruited following UVB irradiation. The adaptor protein STING was necessary for both type I IFN and proinflammatory cytokine expression in the skin. UVB‐irradiated IFNAR‐knockout mice showed increased levels of proinflammatory genes and more severe inflammation by histology, suggesting a protective role for type I IFN.

Conclusion

In wild‐type mice, repeated doses of UVB irradiation induce monocyte‐dependent and PDC‐independent expression of type I IFN together with expression of other proinflammatory cytokines. Induction is dependent on the adaptor protein STING. Surprisingly, studies using IFNAR‐deficient mice revealed that type I IFN protects against UVB irradiation–induced skin inflammation, in part by attenuating proinflammatory cytokine expression and limiting tissue damage.
Asymmetric Aldol Synthesis: Choice of Organocatalyst and Conditions
Authors: Nakashima Erika , Yamamoto Hisashi
Published: November 2016

This is the first report of a successful asymmetric aldol reaction of acetone and 4‐nitrobenzaldehyde catalyzed by only 0.01 mol % of proline tetrazole (5‐pyrrolidin‐2‐yltetrazole). Amazingly, 3 mol % of water as an additive has significantly improved the efficiency of the aldol reaction in terms of yield and enantioselectivity, up to a 0.1‐mol‐scale production (up to 99 % isolated yield, 84 % ee).
Delay‐Dependent Stability Criterion for Discrete‐Time Systems with Time‐Varying Delays
Authors: Hua Changchun , Wu Shuangshuang , Bai Zhenhua , Guan Xinping
Published: November 2016

The stability analysis problem is considered for linear discrete‐time systems with time‐varying delays. A novel summation inequality is proposed, which takes the double summation information of the system state into consideration. The inequality relaxes the recently proposed discrete Wirtinger inequality and its improved version. Based on construction of a suitable Lyapunov‐Krasovskii functional and the novel summation inequality, an improved delay‐dependent stability criterion for asymptotic stability of the systems is derived in terms of linear matrix inequalities. Numerical examples are given to demonstrate the advantages of the proposed method.
Arachidonic acid and lipoxin A4 attenuate alloxan‐induced cytotoxicity to RIN5F cells in vitro and type 1 diabetes mellitus in vivo
Authors: Gundala Naveen K.V. , Naidu Vegi G.M. , Das Undurti N.
Published: November 2016

Objective: We studied whether polyunsaturated fatty acids (PUFAs) can protect rat insulinoma (RIN5F) cells against alloxan‐induced apoptosis in vitro and type 1 diabetes mellitus (type 1 DM) in vivo and if so, mechanism of this beneficial action. Material and Methods: In vitro study was conducted using RIN5F cells while in vivo study was performed in Wistar rats. The effect of PUFAs, cyclo‐oxygenase and lipoxygenase inhibitors, various eicosanoids and PUFAs metabolites: lipoxin A4 (LXA4), resolvin D2 and protectin against alloxan‐induced cytotoxicity to RIN5F cells and type 1 DM was studied. Expression of PDX1, P65 NF‐kB and IKB in RIN5F cells and Nrf2, GLUT2, COX2, iNOS protein levels in the pancreatic tissue and plasma glucose, insulin and tumor necrosis factor‐α and antioxidants, lipid peroxides and nitric oxide were measured. Results: Of all, arachidonic acid (AA) was found to be the most effective against alloxan‐induced cytotoxicity to RIN5F cells and preventing type 1 DM. Both cyclo‐oxygenase and lipoxygenase inhibitors did not block the beneficial actions of AA in vitro and in vivo. Alloxan inhibited LXA4 production by RIN5F cells and in alloxan‐induced type 1 DM Wistar rats. AA‐treatment restored LXA4 levels to normal both in vitro and in vivo. LXA4 protected RIN5F cells against alloxan‐induced cytotoxicity and prevented type 1 DM and restored expression of Nrf2, Glut2, COX2, and iNOS genes and abnormal antioxidants to near normal. Discussion: AA seems to bring about its beneficial actions against alloxan‐induced cytotoxicity and type 1 DM by enhancing the production of LXA4. © 2016 BioFactors, 43(2):251–271, 2017
Serum albumin as a local therapeutic agent in cell therapy and tissue engineering
Authors: Horváthy Dénes B. , Simon Melinda , Schwarz Charlotte M. , Masteling Mariana , Vácz Gabriella , Hornyák István , Lacza Zsombor
Published: November 2016

Albumin is a major plasma protein that has become ubiquitous in regenerative medicine research. As such, many studies have examined its structure and advantageous properties. However, a systematic and comprehensive understanding of albumin's role, capabilities and therapeutic potential still eludes the field. In the present work, we review how albumin is applied in tissue engineering, including cell culture and storage, in vitro fertilization and transplantation. Furthermore, we discuss how albumin's physiological role extends beyond a carrier for metal ions, fatty acids, pharmacons and growth factors. Albumin acts as a bacteriostatic coating that simultaneously promotes attachment and proliferation of eukaryotic cells. These properties with the combination of free radical scavenging, neutrophil activation and as a buffer molecule already make the albumin protein beneficial in healing processes supporting functional tissue remodeling. Nevertheless, recent data revealed that albumin can be synthesized by osteoblasts and its local concentration is raised after bone trauma. Interestingly, by increasing the local albumin concentration in vivo, faster bone healing is achieved, possibly because albumin recruits endogenous stem cells and promotes the growth of new bone. These data also suggest an active role of albumin, even though a specific receptor has not yet been identified. Together, this discussion sheds light on why the extravascular use of the albumin molecule is in the scope of scientific investigations and why it should be considered as a local therapeutic agent in regenerative medicine. © 2016 BioFactors, 43(3):315–330, 2017
Immobilized hematopoietic growth factors onto magnetic particles offer a scalable strategy for cell therapy manufacturing in suspension cultures
Authors: Worrallo Matthew J. , Moore Rebecca L.L. , Glen Katie E. , Thomas Robert J.
Published: November 2016

Hematopoietic therapies require high cell dosages and precise phenotype control for clinical success; scalable manufacturing processes therefore need to be economic and controllable, in particular with respect to culture medium and growth factor (GF) strategy. The aim of this work was to demonstrate the biological function, and integration within scalable systems, of a highly controllable immobilized growth factor (iGF) approach. GFs were biotinylated and attached to streptavidin coated magnetic particles. GF concentration during biotinylation, GF‐biotin ratio, and GF lysine content were shown to control iGF surface concentration and enable predictable co‐presentation of multiple GF on a single bead. Function was demonstrated for immobilized GMCSF, SCF, TPO and IL‐3 in GF dependent cell lines TF‐1 and M‐07e. Immobilized GMCSF (iGMCSF) was analyzed to show sustained activity over 8 days of culture, a 2–3 order of magnitude potency increase relative to soluble factor, and retained functionality under agitation in a micro‐scale stirred tank bioreactor. Further, short exposure to iGMCSF demonstrated prolonged growth response relative to soluble factor. This immobilization approach has the potential to reduce the manufacturing costs of scaled cell therapy products by reducing GF quantities and offers important process control opportunities through separation of GF treatments from the bulk media.
Metabolic engineering of Methanosarcina acetivorans for lactate production from methane
Authors: McAnulty Michael J. , Poosarla Venkata Giridhar , Li Jine , Soo Valerie W. C. , Zhu Fayin , Wood Thomas K.
Published: November 2016

We previously demonstrated anaerobic conversion of the greenhouse gas methane into acetate using an engineered archaeon that produces methyl‐coenzyme M reductase (Mcr) from unculturable microorganisms from a microbial mat in the Black Sea to create the first culturable prokaryote that reverses methanogenesis and grows anaerobically on methane. In this work, we further engineered the same host with the goal of converting methane into butanol. Instead, we discovered a process for converting methane to a secreted valuable product, L‐lactate, with sufficient optical purity for synthesizing the biodegradable plastic poly‐lactic acid. We determined that the 3‐hydroxybutyryl‐CoA dehydrogenase (Hbd) from Clostridium acetobutylicum is responsible for lactate production. This work demonstrates the first metabolic engineering of a methanogen with a synthetic pathway; in effect, we produce a novel product (lactate) from a novel substrate (methane) by cloning the three genes for Mcr and one for Hbd. We further demonstrate the utility of anaerobic methane conversion with an increased lactate yield compared to aerobic methane conversion to lactate. Biotechnol. Bioeng. 2017;114: 852–861. © 2016 Wiley Periodicals, Inc.
Establishing very long‐chain fatty alcohol and wax ester biosynthesis in Saccharomyces cerevisiae
Authors: Wenning Leonie , Yu Tao , David Florian , Nielsen Jens , Siewers Verena
Published: November 2016

Wax esters (WEs) are neutral lipids and can be used for a broad range of commercial applications, including personal care products, lubricants, or coatings. They are synthesized by enzymatic reactions catalyzed by a fatty acyl reductase (FAR) and a wax ester synthase (WS). At present, commercially used WEs are mainly isolated from Simmondsia chinensis (jojoba), but the high extraction costs and limited harvest areas constrain their use. The use of FARs in combination with different WSs to achieve a synthesis of jojoba‐like WEs in bacteria and yeast has been reported previously, but the products were restricted to C28–C36 WEs. These rather short WEs make up only a very small percentage of the total WEs in natural jojoba oil. The synthesis of longer chain WEs (up to C44) in Saccharomyces cerevisiae has so far only been achieved after substrate feeding. Here we identified new routes for producing very long‐chain fatty alcohols (VLCFOHs) up to a chain length of C22 by heterologous expression of a FAR derived from Apis mellifera (AmFAR1) or Marinobacter aquaeolei VT8 (Maqu_2220) in S. cerevisiae and achieved maximum yields of 3.22 ± 0.36 mg/g cell dry weight (CDW) and 7.84 ± 3.09 mg/g CDW, respectively, after 48 h. Moreover, we enabled the synthesis of jojoba‐like WEs up to a chain length of C42, catalyzed by a combination of Maqu_2220 together with the WS from S. chinensis (SciWS) and the S. cerevisiae elongase Elo2p, with a maximum yield of 12.24 ± 3.35 mg/g CDW after 48 h. Biotechnol. Bioeng. 2017;114: 1025–1035. © 2016 Wiley Periodicals, Inc.
Shuttle‐mediated nanoparticle transport across an in vitro brain endothelium under flow conditions
Authors: Falanga Andrea P. , Pitingolo Gabriele , Celentano Maurizio , Cosentino Armando , Melone Pietro , Vecchione Raffaele , Guarnieri Daniela , Netti Paolo A.
Published: November 2016

The blood brain barrier (BBB) represents a challenge in the development of new nano‐delivery systems able to reach the central nervous system (CNS). In order to test the efficacy of these nanocarriers, it is fundamental to use in vitro models that resemble the in vivo cell culture conditions. Here, we demonstrate for the first time the ability of a membranotropic peptide, namely gH625, to transport a cargo—acting as a shuttle—across the BBB layer under flow conditions that mimic the blood flow rate. To this aim, a BBB microfluidic device was designed based on a transparent polyester porous membrane sandwiched between a top and a bottom overlying channel made of poly(methyl methacrylate) (PMMA). Our data clearly indicate that this microfluidic system allows the growth of brain endothelial bEnd.3 cells and the formation of a confluent layer at 7 days of culture that hinders the passage of nanoparticles compared to porous membrane alone. The device was validated at a 5 μL/min working flow rate, where the capability of the model to remain intact after nanoparticle passage was shown. Very interestingly, the decoration with the gH625 peptide enhances the adhesion of nanoparticles to the endothelial layer and the BBB crossing in flow conditions, thus confirming the efficacy of the gH625 as a delivery platform to the brain. Biotechnol. Bioeng. 2017;114: 1087–1095. © 2016 Wiley Periodicals, Inc.
Method of breast reconstruction and the development of lymphoedema
Authors: Lee K-T , Bang S I , Pyon J-K , Hwang J H , Mun G-H
Published: November 2016

Background

Several studies have demonstrated an association between immediate autologous or implant-based breast reconstruction and a reduced incidence of lymphoedema. However, few of these have ocused specifically on whether the reconstruction method affects the development of lymphoedema. The study evaluated the potential impact of breast reconstruction modality on the incidence of lymphoedema.

Methods

Outcomes of women with breast cancer who underwent mastectomy and immediate reconstruction using an autologous flap or a tissue expander/implant between 2008 and 2013 were reviewed. Arm or hand swelling with pertinent clinical signs of lymphoedema and excess volume compared with those of the contralateral side was diagnosed as lymphoedema. The cumulative incidence of lymphoedema was estimated by the Kaplan–Meier method. Clinicopathological factors associated with the development of lymphoedema were investigated by Cox regression analysis.

Results

A total of 429 reconstructions (214 autologous and 215 tissue expander/implant) were analysed; the mean follow-up of patients was 45·3 months. The two groups had similar characteristics, except that women in the autologous group were older, had a higher BMI, and more often had preoperative radiotherapy than women in the tissue expander/implant group. Overall, the 2-year cumulative incidence of lymphoedema was 6·8 per cent (autologous 4·2 per cent, tissue expander/implant 9·3 per cent). Multivariable analysis demonstrated that autologous reconstruction was associated with a significantly reduced risk of lymphoedema compared with that for tissue expander/implant reconstruction. Axillary dissection, a greater number of dissected lymph nodes and postoperative chemotherapy were also independent risk factors for lymphoedema.

Conclusion

The method of breast reconstruction may affect subsequent development of lymphoedema.
Blog construction as an effective tool in biochemistry active learning
Authors: Cubas Rolim Estêvão , Martins de Oliveira Julia , Dalvi Luana T. , Moreira Daniel C. , Garcia Caldas Natasha , Fernandes Lobo Felipe , André Polli Démerson , Campos Élida G. , Hermes‐Lima Marcelo
Published: November 2016

To boost active learning in undergraduate students, they were given the task of preparing blogs on topics of clinical biochemistry. This “experiment” lasted for 12 teaching‐semesters (from 2008 to 2013), and included a survey on the blogs' usefulness at the end of each semester. The survey (applied in the 2008‐2010 period) used a Likert‐like questionnaire with eight questions and a 1‐to‐6 scale, from “totally disagree” to “fully agree.” Answers of 428 students were analyzed and indicated overall approval of the blog activity: 86% and 35% of the responses scored 4‐to‐6 and 6, respectively. Considering the survey results, the high grades obtained by students on their blogs (averaging 8.3 in 2008–2010), and the significant increase in average grades of the clinical biochemistry exam after the beginning of the blog system (from 5.5 in 2007 to 6.4 in 2008–2010), we concluded that blogging activity on biochemistry is a promising tool for boosting active learning. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(3):205–215, 2017.
Validated LC–MS/MS method for simultaneous determination of doxorubicin and curcumin in polymeric micelles in subcellular compartments of MCF‐7/Adr cells by protein precipitation–ultrasonic breaking method
Authors: Wang Jinling , Li Ying , Ma Wenzhuan , Wang Xiaohui , Tu Pengfei
Published: November 2016

A specific and sensitive LC–MS/MS with protein precipitation– ultrasonic breaking method has been developed and validated for simultaneous determination of doxorubicin (DOX) and curcumin (Cur) in DOX and Cur co‐loaded hyaluronic acid–vitamin E succinatemicelles [(DOX + Cur)–polymeric micelles (PMs)] in subcellular compartments of resistant MCF‐7/Adr cells. Sequential extraction of four subcellular protein fractions (cytosolic, membrane/organelle, nucleic and cytoskeleton) was performed directly from MCF‐7/Adr cells after incubation with (DOX + Cur)–PMs. An ultrasonic breaking–methanol precipitation method was used for extraction of the fractions, and the micelle breaking efficiency with methanol was 98.1 and 97.6% for DOX and Cur, respectively. The analytes were analyzed using positive electrospray ionization coupled with multiple reaction monitoring. The calibration curves were linear over a concentration range of 0.5–400 ng/mL for DOX and 2–2000 ng/mL for Cur, and the recovery for the two analytes were >85% with negligible matrix effect. The intra‐day and inter‐day precision was <10.80% and relative error was within ±7.70%. The developed method was successfully applied for subcellular determination of DOX and Cur in MCF‐7/Adr cells. Moreover, Cur and (DOX + Cur)–PMs had a marked promoting effect on the distribution of DOX in the nucleic protein fraction.
Application of a solvent‐free solid injection technique coupled with GC–MS for discrimination between the secondary metabolites of wild and cultivated South Korean medicinal foods
Authors: Truong Lieu T. B. , Abd El‐Aty A. M. , Kim Hyun Jin , Rahman Md. Musfiqur , Kim Sung‐Woo , Shin Ho‐Chul , Shim Jae‐Han
Published: November 2016

Solvent‐free solid injection was applied to differentiate between wild and cultivated South Korean medicinal foods, including dureup (Aralia elata), deodeok (Codonopsis lanceolata) and doraji (Platycodon grandiflorus). A number of compounds were identified in wild and cultivated dureup (53 and 46), deodeok (47 and 51) and doraji (43 and 38). Secondary metabolites, including butanal,2‐methyl‐, β‐caryophyllene, neoclovene, α‐humulene, γ‐curcumene, β‐bisabolene, and phytol, were identified in dureup with significantly (P < 0.05) different amounts between both types. In deodeok, squalene and other main components such as acetic acid, methyl ester, furan‐methyl‐furfural, 2‐furan‐methanol, and 5‐methyl‐furfural, were statistically different between the two types. Doraji has significantly different compounds such as furfural, 5‐methyl‐furfural, 2‐methoxy‐phenol, 2‐methoxy‐4‐(1‐propenyl)‐phenol, and 1‐(4‐hydroxy‐3‐methoxyphenyl)‐2‐propanone. Although we failed to confirm the key compounds, a new compound, namely desaspidinol, was synthesized for the first time and its retention index determined under the experimental conditions. This solventless, easy technique can be used as a simple way to discriminate between wild and cultivated types of medicinal plants via identification of volatile markers or specific fingerprints.
Heterologous expression of the plant cysteine protease bromelain and its inhibitor in Pichia pastoris
Authors: Luniak Nora , Meiser Peter , Burkart Sonja , Müller Rolf
Published: November 2016

Expression of proteases in heterologous hosts remains an ambitious challenge due to severe problems associated with digestion of host proteins. On the other hand, proteases are broadly used in industrial applications and resemble promising drug candidates. Bromelain is an herbal drug that is medicinally used for treatment of oedematous swellings and inflammatory conditions and consists in large part of proteolytic enzymes. Even though various experiments underline the requirement of active cysteine proteases for biological activity, so far no investigation succeeded to clearly clarify the pharmacological mode of action of bromelain. The potential role of proteases themselves and other molecules of this multi‐component extract currently remain largely unknown or ill defined. Here, we set out to express several bromelain cysteine proteases as well as a bromelain inhibitor molecule in order to gain defined molecular entities for subsequent studies. After cloning the genes from its natural source Ananas comosus (pineapple plant) into Pichia pastoris and subsequent fermentation and purification, we obtained active protease and inhibitor molecules which were subsequently biochemically characterized. Employing purified bromelain fractions paves the way for further elucidation of pharmacological activities of this natural product. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:54–65, 2017
Improving Intergroup Relations with Extended Contact among Young Children: Mediation by Intergroup Empathy and Moderation by Direct Intergroup Contact
Authors: Vezzali Loris , Hewstone Miles , Capozza Dora , Trifiletti Elena , Bernardo Gian Antonio Di
Published: November 2016

A correlational study investigated extended contact as a strategy to improve outgroup attitudes and stereotyping and to prepare children for future contact. Additional aims were to investigate when and why the effects of extended contact occur. In particular, intergroup empathy was tested as a mediator and direct contact (i.e. cross‐group friendship) as a moderator of extended contact. Participants were Italian and immigrant elementary school children. Results showed that extended contact was associated with improved intergroup empathy, which, in turn, was associated with more positive outgroup attitudes, stereotypes and behavioural intentions. These effects were significant only among participants with a low or moderate level of direct contact. The theoretical and practical implications of findings are discussed. Copyright © 2016 John Wiley & Sons, Ltd.
Understand legal aspects of game day protests involving the national anthem
Authors: O'Brien Timothy
Published: November 2016

The increasing number of protests during the national anthem by student‐athletes and others associated with athletic contests during the last two months has resulted in a renewed focus on the legal implications of these actions and on the ability to regulate such conduct.
Reduce liability, boost safety of team road trips by reviewing policies, procedures
Authors: McCarthy Claudine
Published: November 2016

It's every athletics administrator's worst nightmare. A vehicle carrying student‐athletes, coaches, and staff to an away competition winds up in an accident, resulting in injuries or even deaths.
Anti‐HSV‐1 and HSV‐2 Flavonoids and a New Kaempferol Triglycoside from the Medicinal Plant Kalanchoe daigremontiana
Authors: Ürményi Fernanda Gouvêa Gomes , Saraiva Georgia do Nascimento , Casanova Livia Marques , Matos Amanda dos Santos , de Magalhães Camargo Luiza Maria , Romanos Maria Teresa Villela , Costa Sônia Soares
Published: November 2016

Kalanchoe daigremontiana (Crassulaceae) is a medicinal plant native to Madagascar. The aim of this study was to investigate the flavonoid content of an aqueous leaf extract from K. daigremontiana (Kd), and assess its antiherpetic potential. The major flavonoid, kaempferol 3‐O‐β‐d‐xylopyranosyl‐(1 → 2)‐α‐l‐rhamnopyranoside (1), was isolated from the AcOEt fraction (Kd‐AC). The BuOH‐soluble fraction afforded quercetin 3‐O‐β‐d‐xylopyranosyl‐(1 → 2)‐α‐l‐rhamnopyranoside (2) and the new kaempferol 3‐O‐β‐d‐xylopyranosyl‐(1 → 2)‐α‐l‐rhamnopyranoside‐7‐O‐β‐d‐glucopyranoside (3), named daigremontrioside. The crude extract, Kd‐AC fraction, flavonoids 1 and 2 were evaluated using acyclovir‐sensitive strains of HSV‐1 and HSV‐2. Kd‐AC was highly active against HSV‐1 (EC50 = 0.97 μg/ml, SI > 206.1) and HSV‐2 (EC50 = 0.72 μg/ml, SI > 277.7). Flavonoids 1 and 2 showed anti‐HSV‐1 (EC50 = 7.4 μg/ml; SI > 27 and EC50 = 5.8 μg/ml; SI > 8.6, respectively) and anti‐HSV‐2 (EC50 = 9.0 μg/ml; SI > 22.2 and EC50 = 36.2 μg/ml; SI > 5.5, respectively) activities, suggesting the contribution of additional substances to the antiviral activity.
Comparative expression analysis of hypoxia‐inducible factor‐alpha and its natural occurring antisense in breast cancer tissues and adjacent noncancerous tissues
Authors: Tasharrofi Behnoosh , Soudyab Mohammad , Nikpayam Elahe , Iranpour Mostafa , Mirfakhraie Reza , Sarrafzadeh Shaghayegh , Geranpayeh Lobat , Azargashb Eznollah , Sayad Arezou , Ghafouri‐Fard Soudeh
Published: November 2016

Hypoxia‐inducible factors (HIFs) have been shown to be upregulated in tumor tissues and linked with tumor progression and metastasis in breast cancer. Among regulatory mechanisms for HIF expression is a natural occurring antisense named aHIF, which has been shown to be overexpressed in breast cancer and influence the level of the HIF‐1α transcript. In the present study, we analyzed the expression of HIF‐1α and aHIF in breast cancer tissues versus adjacent noncancer tissues (ANCTs) in relation with the clinical and biological behavior of the tumors. aHIF has been shown to be expressed in 67.4% of invasive ductal carcinoma samples, while none of ANCTs showed its expression. HIF‐1α has been expressed in all of tumors and 90% of ANCTs. Comparison of HIF‐1α expression level between tumor and ANCT tissues showed a total upregulation in tumor samples. No statistically significant association has been found between the level of HIF‐1α expression in tumor samples and clinicopathologic and demographic characteristics such as age, tumor size, estrogen receptor status, progesterone receptor status, HER2/neu expression level, lymph node status, histological grade, and stage except for a weak correlation between HIF‐1α expression and Ki‐67 status. Besides, we could not detect any significant correlation between relative expression of HIF‐1α and aHIF in tumor samples. Collectively, these data suggest that aHIF overexpression can be used as a potential biomarker in breast cancer. However, further studies are needed for the evaluation of its mechanism of action in regulation of HIF‐1α expression in different pathological conditions. HIF‐1α overexpression results in the upregulation of several genes that participated in cancer‐associated pathways such as proliferation, angiogenesis, and glucose metabolism. We showed that HIF‐1α is upregulated in breast tumor samples compared with adjacent noncancerous tissues. Its expression has been associated with Ki‐67 status. Its natural occurring antisense is only expressed in tumor tissues. Thus, it can be used as a potential biomarker in breast cancer.
RUNX3 inhibits laryngeal squamous cell carcinoma malignancy under the regulation of miR‐148a‐3p/DNMT1 axis
Authors: Jili Su , Eryong Lu , Lijuan Lu , Chao Zhang
Published: November 2016

Laryngeal squamous cell carcinoma (LSCC) is a highly aggressive malignant cancer and accounts for 1% to 2% of all malignancies diagnosed worldwide. Runt‐related transcription factor 3 (RUNX3), an important tumor suppressor, is known to related to lymph node metastasis and the development of LSCC. However, the biological roles and potential mechanisms RUNX3 expression was not well understood. In this study, we reported that the RUNX3 was significantly downregulated and highly methylated in LSCC compared with their matched normal. The enforced expression of RUNX3 inhibited LSCC cell migration, invasion, and proliferation, whereas the inhibition of RUNX3 did the opposite. We identified that RUNX3 was regulated by miR‐148a‐3p and found that the expression level of miR‐148‐3p was significantly decreased and positively related with the expression of RUNX3 in LSCC. We also identified that DNA methyltransferase enzyme DNA (cytosine‐5‐)‐methyltransferase 1 (DNMT1) was targeted by miR‐148a‐3p in LSCC. The knockdown of DNMT1 promoted the expression of RUNX3 and inhibited migration, invasion, and proliferation in LSCC cells. In summary, our study demonstrated that miR‐148a‐3p may regulate RUNX3 expression through the modulation of DNMT1‐dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC. LSCC is a highly aggressive malignant cancer and accounts for 1% to 2% of all malignancies diagnosed worldwide. In this study, we reported that RUNX3, an important tumor suppressor, was significantly downregulated and highly methylated in LSCC compared with their matched normal. The overexpression of RUNX3 inhibited LSCC cell migration, invasion, and proliferation, whereas the inhibition of RUNX3 did the opposite. Moreover, RUNX3 was regulated by miR‐148a‐3p, which targeted DNA methyltransferase enzyme DNMT1 in LSCC cells. Therefore, miR‐148a‐3p may regulate RUNX3 expression through the modulation of DNMT1‐dependent DNA methylation in LSCC, providing a novel target and a potential therapeutic pathway against LSCC.
Improving the Binding Affinity of in‐Vitro‐Evolved Cyclic Peptides by Inserting Atoms into the Macrocycle Backbone
Authors: Wilbs Jonas , Middendorp Simon J. , Heinis Christian
Published: November 2016

Cyclic peptides binding to targets of interest can be generated efficiently with powerful in vitro display techniques, such as phage display or mRNA display. The cyclic peptide libraries screened with these methods are generated by altering in a combinatorial fashion the amino acid sequence of the peptides, the number of amino acids in the macrocycle rings, and the cyclization chemistry. A structural element that cannot easily be varied in the cyclic peptides is the backbone, which is built from amino acids, each of which contributes three atoms to the macrocyclic ring structure. Here, we proposed to improve the affinity of a phage‐selected bicyclic peptide inhibitor of coagulation factor XII (FXII) by screening variants with one or two carbon atoms inserted into different positions of the backbone, and thus tapping into a structural space that was not sampled by phage display. Two mutants showed 4.7‐ and 2.5‐fold improved Ki values. The better one blocked FXII with a Ki of 1.5±0.1 nm and inhibited activation of the intrinsic coagulation pathway (EC2x 1.7 μm). The strategy of ring size variation by one or several atoms should be generally applicable for the affinity maturation of in‐vitro‐evolved cyclic peptides.
Efficient Conjugation of Oligosaccharides to Polymer Particles through Furan/Maleimide Diels–Alder Reaction: Application to the Capture of Carbohydrate‐Binding Proteins
Authors: Petrelli Antoine , Samain Eric , Pradeau Stéphanie , Halila Sami , Fort Sébastien
Published: November 2016

Glycan–protein interactions play a crucial role in physiological and pathological events. Hence, improving the isolation of carbohydrate‐binding proteins (i.e., lectins and anti‐glycan antibodies) from complex media might not only lead to a better understanding of their function, but also provide solutions for public health issues, such as water contamination or the need for universal blood plasma. Here we report a rapid and efficient method for producing carbohydrate‐based affinity adsorbents combining enzymatic synthesis and metal‐free click chemistry. Both simple and complex glycans (maltose, blood group antigens A, B, and H) were readily modified by the addition of a furyl group at the reducing end without the need for protecting groups and were then efficiently conjugated to maleimide‐activated Sepharose particles through Diels–Alder cycloaddition. These neoglycoconjugates showed high efficiency for the purification of lectins (concanavalin A and Ulex europaeus agglutinin), as well as for the capture of anti‐A and anti‐B blood group antibodies, opening new prospects for glycoproteomics and for the development of universal blood plasma.
FOXP1 enhances tumor cell migration by repression of NFAT1 transcriptional activity in MDA‐MB‐231 cells
Authors: Oskay Halacli Sevil
Published: November 2016

Until now, forkhead box P1 (FOXP1) has been identified as a tumor suppressor in several correlation studies in breast cancer. Although FOXP1 is defined as a transcriptional repressor that interacts with other transcription factors in various mechanistic studies, there is no study that explains its repressor functions in breast cancer biology. This study demonstrated the repressor function of FOXP1 on nuclear factor of activated T cells (NFAT1) and the migratory effect of this repression in MDA‐MB‐231 breast cancer cells. Co‐immunoprecipitation experiments were performed for the investigation of protein–protein interaction between two transcription factors. Protein–protein interaction on DNA was investigated with EMSA and transcriptional effects of FOXP1 on NFAT1, luciferase reporter assay was performed. Wound healing assay was used to analyze the effects of overexpression of FOXP1 on tumor cell migration. This study showed that FOXP1 has protein–protein interaction with NFAT1 on DNA and enhances breast cancer cell migration by repressing NFAT1 transcriptional activity and FOXP1 shows oncogenic function by regulating breast cancer cell motility.
Comparison of transradial and transfemoral access in patients undergoing percutaneous coronary intervention for complex coronary lesions
Authors: Koifman Edward , Gaglia Michael A. , Escarcega Ricardo O. , Bernardo Nelson L. , Lager Robert A. , Gallino Robert A. , Ben‐Dor Itsik , Pichard Augusto D. , Baker Nevin C. , Lipinski Michael J. , Kiramijyan Sarkis , Negi Smita I. , Torguson Rebecca , Gai Jiaxiang , Suddath William O. , Satler Lowell F. , Waksman Ron
Published: November 2016

Objective

Comparison of transradial versus transfemoral access for complex percutaneous coronary intervention (PCI) with regard to both complications and long‐term outcomes.

Background

Radial access has been shown to confer superior results in patients undergoing PCI, especially in patients with acute coronary syndromes. However, radial access has limitations of sheath and device size, which may increase procedure time and result in inferior outcomes.

Methods

Patients undergoing PCI for complex lesions, defined as type C according the ACC/AHA classification system, were included in this study. Propensity matching was performed to adjust for differences in baseline characteristics. Transradial patients were then compared to transfemoral patients in regard to procedural, in‐hospital, and 6‐month outcomes.

Results

Among 2142 patients with 2591 lesions treated, 1876 had femoral access and 267 had radial access. Radial access patients were more likely to be male (75% vs. 66%, P = 0.003) and less likely to present with acute myocardial infarction (27% vs. 42%, P < 0.001). Procedural characteristics demonstrated lower use of heparin in the femoral group (17% vs. 73%, P < 0.001) with similarly low use of glycoprotein inhibitors (5.6% vs. 3.4%, P = 0.14). Patients in the femoral group had higher rates of transfusions (3.7% vs. 0%, P = 0.004) and vascular complications (1.7% vs. 0%, P = 0.03). Following propensity matching, there was no difference in mid‐term outcomes between radial and femoral groups.

Conclusions

In patients with complex coronary lesions undergoing PCI, the radial approach demonstrates similar mid‐term outcomes as the femoral approach with a potentially lower rate of complications. © 2016 Wiley Periodicals, Inc.
Safety and efficacy of rotational atherectomy for the treatment of undilatable underexpanded stents implanted in calcific lesions
Authors: Ferri Luca A. , Jabbour Richard J. , Giannini Francesco , Benincasa Susanna , Ancona Marco , Regazzoli Damiano , Mangieri Antonio , Montorfano Matteo , Colombo Antonio , Latib Azeem
Published: November 2016

Objectives

Coronary stent underexpansion is a known risk factor for in‐stent restenosis and stent thrombosis. There are limited options once noncompliant balloons have failed to achieve optimal stent expansion. Excimer Laser Coronary Angioplasty with contrast medium injection is one possibility, but not readily available. Rotational atherectomy is an alternative, and has been described in case reports, but concerns exist regarding safety.

Methods

All consecutive patients undergoing rotational atherectomy for symptomatic in‐stent restenosis due to stent underexpansion resistant to noncompliant balloon postdilatation between January 2005 and December 2015 were analysed.

Results

A total of 16 patients underwent treatment during the study period and the procedure was successful in 14 cases (87.5%). The mean postprocedural minimal lumen diameter increased by 2.3 ± 0.8 mm and percentage diameter stenosis decreased from 82.17% ± 17.2% to 11.9% ± 9.1%. Intraprocedural complications occurred in two patients (burr entrapment successfully managed percutaneously and periprocedural myocardial infarction). At 1‐year follow‐up, the incidence of target lesion revascularisation was 13.3% (2 out of 15 patients), and one patient died from noncardiac death.

Conclusion

In this small series of underexpanded stents, rotational atherectomy was an effective treatment for resistant stent underexpansion with acceptable outcomes. © 2016 Wiley Periodicals, Inc.
Coronary artery spatial distribution of chronic total occlusions: Insights from a large US registry
Authors: Garcia Santiago , Alraies M. Chadi , Karatasakis Aris , Yannopoulos Demetris , Karmpaliotis Dimitri , Alaswad Khaldoon , Jaffer Farouc A. , Yeh Robert W. , Patel Mitul P. , Bahadorani John , Karacsonyi Judit , Kalsaria Pratik , Danek Barbara , Banerjee Subhash , Brilakis Emmanouil S.
Published: November 2016

Objective

To assess the spatial distribution of chronic total occlusions (CTOs) within the coronary arteries and describe procedural strategies and outcomes during CTO percutaneous coronary intervention (PCI).

Background

Acute occlusions due to plaque rupture tend to cluster within the proximal third of the coronary artery.

Methods

We examined the clinical and procedural characteristics of 1,348 patients according to lesion location within the coronary tree.

Results

A total of 1,369 lesions in 1,348 patients (mean age 66 ± 10 years, 85% male) were included. CTO PCI of proximal segments (n = 633, 46%) was more common than of mid (n = 557, 41%) and distal segments (n = 179, 13%). Patients undergoing CTO PCI of proximal segments were more likely to be smokers (P < 0.01), have prior coronary artery bypass graft surgery (P = 0.03) and lower ejection fraction (P = 0.04). CTOs occurring in proximal segments had longer length (P <0.01), proximal cap ambiguity (P < 0.01), and moderate/severe calcification (P < 0.01) compared to mid or distally located CTOs. Interventional collaterals were more often present in CTO PCI of proximal segments (64%, 53%, 56%, P < 0.01) consistent with the higher use of retrograde approach (47%, 33%, 37%, P < 0.01) relative to antegrade wire escalation (67%, 82%, 82%, P < 0.01). Procedural complexity was higher in CTO PCI of proximal segments (vs. mid and distal): contrast volume= 275 ml (200–375), 260 ml (200–350), 250 ml (175–350), P = 0.01; fluoroscopy time 53 minutes (32–83), 39 minutes (24–65), 40 minutes (22–72), P < 0.01. However, procedural success (87%, 90%, 85%, P = 0.1), technical success (89%, 91%, 88%, P = 0.24), and complications rates (2.8%, 2.5%, 2.2%, P = 0.88) were not different.

Conclusions

The most common target vessel location for CTO PCI is the proximal coronary segment. PCI of proximal occlusions is associated with adverse clinical and angiographic characteristics and often requires use of the retrograde approach, but can be accomplished with high procedural and technical success and low complication rates. © 2016 Wiley Periodicals, Inc.
Closure of very large PDA with pulmonary hypertension: Initial clinical case‐series with the new Occlutech® PDA occluder
Authors: Lehner Anja , Ulrich Sarah , Happel Christoph M. , Fischer Marcus , Kantzis Marinos , Schulze‐Neick Ingram , Haas Nikolaus A.
Published: November 2016

Objective

We report our recent experience with the new Occlutech® PDA Occluder, which seems suitable especially for closure of large patent ductus arteriosus (PDA) sizes with elevated pulmonary artery pressure (PAH).

Background

Although considered a standard procedure in pediatric cath labs, closure of very large PDA (PDA/Aorta ratio > 0.5) and PAH remains a challenge.

Methods/Results

The Occlutech® PDA occluder was used in eight children and adolescents (age 4–16, median 10.75 years), body weight from 14 to 54 kg (median 21 kg) with very large PDA and PAH: PDA diameter was 5–13 mm (median 10 mm); PAP/Aortic pressure ratio was 0.58–0.96. Sufficient occlusion of the PDA was documented by angiography and/or echocardiography in all cases. Four different device sizes were selected: Minimal diameter 6–14 mm (median 12 mm), length 6.3–16 mm (median 14 mm), aortic disc 13–24 mm (median 20 mm), size at the pulmonary end 8–18 mm (median 15 mm). A balloon test occlusion of the PDA was performed in five patients in order to evaulate the decline of pulmonary artery pressure or to delineate the exact anatomy of the PDA. Before release of the device, a careful “wiggle manoevre” was performed to assess the stability of the implanted device and to proof the inability to embolize to the aorta.

Conclusions

With the new Occlutech® PDA Occluder closure of very large PDAs and PAH is feasible and efficient. The wider pulmonary artery end of the Occlutech® PDA Occluder offers enhanced stability and reduces the risk of embolization. © 2016 Wiley Periodicals, Inc.
Feasibility and 1‐Year outcomes of subintimal revascularization with supera® stenting of long femoropopliteal occlusions in critical limb ischemia
Authors: Palena Luis M. , Diaz‐Sandoval Larry J. , Sultato Enrico , Brigato Cesare , Candeo Alessandro , Brocco Enrico , Manzi Marco
Published: November 2016

Background

Stent‐based revascularization of long femoro‐popliteal (FP) lesions has been mainly studied in claudicants and compromised by restenosis and stent fractures. The Supera® stent's biomimetic design allows enhanced fracture resistance. Data for Supera® stenting to treat long chronic total occlusions (CTOs) in patients with critical limb ischemia (CLI), are scarce.

Objective

To assess long‐term outcomes of subintimal revascularization with Supera® stenting, for long FP CTOs in patients with CLI.

Methods

Prospective, single‐center, single‐arm study of 34 consecutive CLI patients with FP TASC C and D CTOs, who underwent Supera® stenting after subintimal crossing. Primary efficacy endpoint was 1‐year patency and freedom from target lesion revascularization (TLR). Primary safety endpoint was the composite rate of freedom from death from any cause, major amputations, and TLR at a year. Secondary endpoints were stent integrity, clinical improvement, amputation free‐survival, quality of life, and cost‐efficiency.

Results

Mean lesion length was 27.9 ± 10.2 cm. Acute technical success was 100%. Primary patency was 94.1%. Freedom from TLR was 97.1%. Limb salvage was 100%. Clinical improvement was observed in 100% of patients: TCPO2 increased from 12.7 ± 6.2 to 54.8 ± 8.4 mm Hg (p < 0,0001); and 100% of patients experienced a shift in Rutherford to class 0 (p < 0.0001). There were no stents fractures. Amputation free‐survival was 82.4%.

Conclusions

Subintimal revascularization with Supera® stenting in CLI patients with long FP occlusions, is feasible and superior to validated efficacy performance goals. Larger multicenter studies are needed to validate the safety and efficacy of this novel alternative approach. © 2016 Wiley Periodicals, Inc.