bluuebunny/crossref_metadata_2025 huggingface dataset:
A new species ofAllogenus(Tricladida, Maricola, Uteriporidae) from South Georgia, Sub-Antarctica
Authors: Volonterio Odile , Brewin Paul E.
Published: December 2013

South Georgia is a remote sub-Antarctic island, considered a marine biodiversity ‘hotspot’ in the Southern Ocean. During a survey along the north coast of South Georgia several marine planarians were found. One of the specimens was a new species ofAllogenus(Uteriporidae), which is described here asAllogenus sluysisp. nov. The new species has the characteristics of the genus and can be distinguished from the type and only known species,Allogenus kerguelensis, by its smaller size, blackish-brown pigmentation, presence of three retinal cells in each eye cup, position of its testes half-way between the ventral and dorsal body surfaces or at a slightly more ventral position, and by having an ejaculatory duct that opens centrally at the tip of the penis papilla. The presently known geographical distribution ofAllogenus, the heterogeneous marine planarian species composition in South Georgia, and the distribution of these species in this region are in agreement with a previously proposed vicariance hypothesis, albeit that dispersal cannot be ruled out.
Effects of CPAP Treatment Interruption Due to Disasters: Patients with Sleep-disordered Breathing in the Great East Japan Earthquake and Tsunami Area
Authors: Mito Fumitaka , Nishijima Tsuguo , Sakurai Shigeru , Kizawa Tetsuya , Hosokawa Keisuke , Takahashi Susumu , Suwabe Akira , Akasaka Hiroshi , Kobayashi Sei-ichiro
Published: December 2013

Introduction

The 2011 Great East Japan Earthquake caused major disruptions in the provision of health care, including that for patients with sleep-disordered breathing (SDB) using a nasal continuous positive airway pressure (nCPAP) device. This study investigated the ability of SDB patients to continue using the nCPAP device in the weeks immediately following the earthquake, whether inability to use the nCPAP device led to symptom relapse, and measures that should be taken to prevent disruptions in nCPAP therapy during future disasters.

Hypothesis

If nCPAP devices cannot be used during disasters, SDB patients’ health will be affected negatively.

Methods

Within 14 days of the disaster, 1,047 SDB patients completed a questionnaire that collected data regarding ability to use, duration of inability to use, and reasons for inability to use the nCPAP device; symptom relapse while unable to use the nCPAP device; ability to use the nCPAP device use at evacuation sites; and recommendations for improvement of the nCPAP device.

Results

Of the 1,047 patients, 966 (92.3%) had been unable to use the nCPAP device in the days immediately following the earthquake. The most common reason for inability to use the nCPAP device was power failure, followed by anxiety about sleeping at night due to fear of aftershocks, involvement in disaster-relief activities, loss of the nasal CPAP device, and fear of being unable to wake up in case of an emergency. Among the 966 patients, 242 (25.1%) had experienced relapse of symptoms, the most common of which was excessive daytime sleepiness (EDS), followed by insomnia, headache, irritability, and chest pain.

Conclusion

Developing strategies for the continuation of nCPAP therapy during disasters is important for providing healthy sleeping environments for SDB patients in emergency situations.

MitoF, NishijimaT, SakuraiS, KizawaT, HosokawaK, TakahashiS, SuwabeA, AkasakaH, KobayashiS. Effects of CPAP treatment interruption due to disasters: patients with sleep-disordered breathing in the Great East Japan Earthquake and tsunami area. Prehosp Disaster Med. 2013;28(6):547-555.
Keratan sulfate expression in microglia is diminished in the spinal cord in experimental autoimmune neuritis
Authors: Matsui H , Ohgomori T , Natori T , Miyamoto K , Kusunoki S , Sakamoto K , Ishiguro N , Imagama S , Kadomatsu K
Published: December 2013

Experimental autoimmune neuritis (EAN) is an animal model of Guillain–Barré syndrome, an inflammatory demyelination disease of the peripheral nervous system. Although this disease has been extensively studied on peripheral nerves, the pathology of the central nervous system has not been fully understood. Previous studies demonstrate that expression of keratan sulfate (KS), the sugar chain of proteoglycan, is associated with activated microglia/macrophages accumulated after neuronal injuries. Unexpectedly, we found here that KS is rather diminished in rat EAN. KS was restrictively expressed in microglia in the spinal cord of normal rats. KS was positive in 50% microglia in the ventral horn and 20% in the dorsal horn. In EAN, microglia increased in number and expressed the activation marker CD68, but KS expression was abolished. Concomitantly, pro-inflammatory cytokines, i.e., interferon (IFN)-γ, interleukin (IL)-1β, and tumor necrosis factor (TNF)-α, were increased in the spinal cord of EAN rats, whereas anti-inflammatory cytokines, such as IL-4 and IL-10, were decreased. In addition, silencing of KSGal6ST attenuated KS expression on the primary cultured microglia and upregulated expression of some activation markers (TNF-α, IL-1β, and iNOS) under the stimulation with lipopolysaccharide and IFN-γ. This study demonstrates for the first time a close association of EAN and disappearance of KS on microglia. KS expression could be a useful marker to evaluate the status of polyneuropathy.
Monitoring Cerebral Oxygenation during Balloon Occlusion with Multichannel NIRS
Authors: Rummel Christian , Zubler Christoph , Schroth Gerhard , Gralla Jan , Hsieh Kety , Abela Eugenio , Hauf Martinus , Meier Niklaus , Verma Rajeev K , Andres Robert H , Nirkko Arto C , Wiest Roland
Published: December 2013

We report on oxygenation changes noninvasively recorded by multichannel continuous-wave near infrared spectroscopy (CW-NIRS) during endovascular neuroradiologic interventions requiring temporary balloon occlusion of arteries supplying the cerebral circulation. Digital subtraction angiography (DSA) provides reference data on the site, timing, and effectiveness of the flow stagnation as well as on the amount and direction of collateral circulation. This setting allows us to relate CW-NIRS findings to brain specific perfusion changes. We focused our analysis on the transition from normal perfusion to vessel occlusion, i.e., before hypoxia becomes clinically apparent. The localization of the maximal response correlated either with the core (occlusion of the middle cerebral artery) or with the watershed areas (occlusion of the internal carotid artery) of the respective vascular territories. In one patient with clinically and angiographically confirmed insufficient collateral flow during carotid artery occlusion, the total hemoglobin concentration became significantly asymmetric, with decreased values in the ipsilateral watershed area and contralaterally increased values. Multichannel CW-NIRS monitoring might serve as an objective and early predictive marker of critical perfusion changes during interventions—to prevent hypoxic damage of the brain. It also might provide valuable human reference data on oxygenation changes as they typically occur during acute stroke.
Characterization of the Ubiquitin-Modified Proteome Regulated by Transient Forebrain Ischemia
Authors: Iwabuchi Masahiro , Sheng Huaxin , Thompson JWill , Wang Liangli , Dubois Laura G , Gooden David , Moseley MArthur , Paschen Wulf , Yang Wei
Published: December 2013

Ubiquitylation is a posttranslational protein modification that modulates various cellular processes of key significance, including protein degradation and DNA damage repair. In animals subjected to transient cerebral ischemia, ubiquitin-conjugated proteins accumulate in Triton-insoluble aggregates. Although this process is widely considered to modulate the fate of postischemic neurons, few attempts have been made to characterize the ubiquitin-modified proteome in these aggregates. We performed proteomics analyses to identify ubiquitylated proteins in postischemic aggregates. Mice were subjected to 10 minutes of forebrain ischemia and 4 hours of reperfusion. The hippocampi were dissected, aggregates were isolated, and trypsin-digested after spiking with GG-BSA as internal standard. K- ε-GG-containing peptides were immunoprecipitated and analyzed by label-free quantitative liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis. We identified 1,664 peptides to 520 proteins containing at least one K- ε-GG. Sixty-six proteins were highly ubiquitylated, with 10 or more K- ε-GG peptides. Based on selection criteria of greater than fivefold increase and P<0.001, 763 peptides to 272 proteins were highly enriched in postischemic aggregates. These included proteins involved in important neuronal functions and signaling pathways that are impaired after ischemia. Results of this study could serve as an important platform to uncover the mechanisms linking insoluble ubiquitin aggregates to the functions of postischemic neurons.
Targeting microRNA-23a to inhibit glioma cell invasion via HOXD10
Authors: Hu Xing , Chen Dan , Cui Yanhui , Li Zhiyuan , Huang Jufang
Published: December 2013

Glioma is the most frequent primary brain tumor. Recently, the upregulation of microRNA (miR)-23a was found to be associated with glioma, but the molecular mechanism by which miR-23a promotes glioma growth remains to be unveiled. In the present study, we found that miR-23a was significantly upregulated in glioma tissues compared to their matched adjacent tissues. miR-23a was also highly expressed in glioma cell lines SHG44, U251 and U87 cells. Moreover, we identified homeobox D10 (HOXD10) as a novel target for miR-23a. The expression of HOXD10 was significantly reduced in glioma tissues and cell lines and miR-23a negatively regulates the protein expression of HOXD10 in U251 and U87 cells. We further showed that miRNA-23a promoted U251 and U87 cell invasion, at least partially, by directly targeting HOXD10 and further modulating MMP-14. These findings suggest that miR-23a may serve as a promising therapeutic target for glioma.
Active Anti-erosion Protection Strategy in Tamarisk (Tamarix aphylla)
Authors: Han Zhiwu , Yin Wei , Zhang Junqiu , Niu Shichao , Ren Luquan
Published: December 2013

Plants have numerous active protection strategies for adapting to complex and severe environments. These strategies provide endless inspiration for extending the service life of materials and machines. Tamarisk (Tamarix aphylla), a tree that thrives in raging sandstorm regions, has adapted to blustery conditions by evolving extremely effective and robust erosion resistant characteristics. However, the relationships among its surface cracks, internal histology and biomechanics, such as cracks, rings, cells, elasticity modulus and growth stress, which account for its erosion resistance, remain unclear. This present study reveals that the directionally eccentric growth rings of tamarisk, which are attributed to reduced stress and accelerated cell division, promote the formation of surface cracks. The windward rings are more extensive than the leeward side rings. The windward surfaces are more prone to cracks, which improves erosion resistance. Our data provide insight into the active protection strategy of the tamarisk against wind–sand erosion.
Synchrotron Radiation for Diagnosis of Skin Conditions
Authors: Cócera M. , Rodríguez G. , Rubio L. , Fernández E. , Barbosa-Barros L. , Labrador A. , Sabés M. , de la Maza A. , López O.
Published: December 2013

The skin acts as a physical barrier at the interface with the external environment. This barrier is designed to protect the organism against insults, including desiccant, mechanical, chemical, and microbial damage. It is primarily composed of three layers: the epidermis, dermis, and hypodermis. The epidermis, which contains the stratum corneum (SC), works as a barrier, preventing water loss from the body and protecting it from the environment. The dermis contains fibroblasts as the predominant cell type within a matrix of structural proteins (collagen or elastin), proteoglycans, nervous fibres and sebaceous glands. The hypodermis acts as a thermal and mechanical insulator. Skin contains non-crystalline material, such as collagen, SC lipids and fat, with characteristic scattering patterns that are altered under specific conditions. These changes can be useful for understanding the structure and state of the tissue.
Section Editorial–Dermal Formulation Development
Authors: Brown M. B.
Published: December 2013

Since the mid-1990s, pharmaceutical R&D productivity has experienced what at best can be described as a downturn. From 1998 to 2008 the number of new molecular entities has been in general decline whereas attrition rates, development times and expenditure have all increased
The Effect of Disulphide Bond Disruption on the Barrier Integrity of the Human Nail
Authors: Baraldi A. , Khengar R. H. , Murdan S. , Traynor M. J. , Jones S. A. , Brown M. B.
Published: December 2013

Onychomycosis is the fungal infection of the nail and accounts for approximately 50% of all nail disorders [1]. The prevalence of onychomycosis is increasing and currently up to 10 % of people in the UK and US are reported to be affected by this condition [2,3]. It can be both physically and psychologically debilitating during the most progressive stages of the disease [4,5]. Fungi (most frequently Trichophyton rubrum and Trichophyton mentagrophytes), moulds (Scytalidium spp, Scopulariopsis spp, Fusarium spp, Acremonium spp, Onychocola canadensis) and yeasts (Candida spp) can be co-resident in a diseased nail and hence not every nail can be considered as being populated by the same community of species.
Pickering Emulsions Stabilised by Biodegradable Particles Offer a Double Level of Controlled Delivery of Hydrophobic Drugs
Authors: Laredj-Bourezg F. , Bolzinger M.-A. , Pelletier J. , Rovere M.-R. , Smatti B. , Chevalier Y.
Published: December 2013

Pickering emulsions are stabilised by solid particles in place of surfactants. Their “surfactant-free” character makes them attractive for cosmetic and pharmaceutical applications because classical emulsions comprising surfactants often show adverse effects such as irritancy, hemolytic behavior, etc. Thus, biocompatible and biodegradable Pickering emulsions would provide an obvious benefit over current technology.
Foam – A Unique Topical Drug Delivery System
Authors: Tamarkin D.
Published: December 2013

Topical therapy is one of the foundations of a dermatologist's therapeutic tools as it is used for symptomatic relief, control, or cure of the underlying disease. The vehicles used to deliver topical therapy can considerably influence drug performance. The vehicle can have direct effects on disease; it can affect the delivery of the active agent, its physical appearance and contribute to sensory properties which affect compliance. While semi-solid compositions such as creams, lotions, gels and ointments are commonly used by consumers, new forms are desirable in order to achieve better control of the application, improve skin absorption and maintain or bestow the beneficial properties of such topical products.
Making Sense of the Cosmeceuticals Concept
Authors: Fernandes D.
Published: December 2013

It is well-known that conventional cosmetics should have no physiological effect on the skin and should make no changes. Theoretically they should only moisturise the stratum corneum and in that way make wrinkles and fine lines less apparent. They should have no effects deeper than that level.
Discussing Cosmetics Through a Functional Scope – A Potential Contribution to Facilitate Decisions and Procedures
Authors: Rodrigues L. M.
Published: December 2013

According to the last published report by Cosmetics Europe, The Personal Care Association, previously known as Colipa, the European Union (EU) cosmetic market represents over €66 billion, making it by far the most important consumer market in the world. It is responsible for an estimated employment (direct and indirect) of 1.7 million people, consisting of more than 4,000 manufacturers of which more than two thirds are small medium enterprises (SMEs).
The Application of Gaussian Processes in the Prediction of Permeability Across a Polydimethlysiloxane Membrane
Authors: Moss G. P. , Sun Y. , Davey N. , Adams R. G. , Wilkinson S. C. , Gullick D. R.
Published: December 2013

Polydimethylsiloxane (PDMS) silicone membranes, such as Silastic®, have been used widely in place of mammalian tissue in the determination of percutaneous absorption. While many experiments have shown correlations between the permeability across both membranes, Moss et al. demonstrated in a systematic study that PDMS membranes tend to exhibit greater permeability than mammalian skin, and that the relationship between permeability across both membranes was not found when the lipophilicity of the penetrant was greater than 3. Further, it was shown previously that when five commonly used physicochemical descriptors were applied to human, pig and rodent membranes they cannot represent the main characteristics of the PDMS dataset when using Gaussian Process (GP) regression to predict skin permeability. However, the previous study in which this process was modelled employed a small dataset (n=19, as part of the wider aims of that work to investigate the effect of dataset construction on model quality).
Section Editorial–Skin Protection
Authors: Hafeez F. , Maibach H. I.
Published: December 2013

Human skin provides a partially effective barrier against potentially toxic chemicals. Much of this barrier protection can be attributed to the stratum corneum (SC), the uppermost and nonviable layer of the epidermis because the rate-limiting step for the penetration of most substances is diffusion through the SC.
Human Scalp Decontamination After Chemical Warfare Agent Exposure
Authors: Rolland P. , Bolzinger M.-A. , Josse D. , Briançon S.
Published: December 2013

The occurrence of exposure to chemical warfare agents is not limited to the military. The Tokyo subway attack in 1995 showed that civilians could also be exposed to highly toxic nerve agents. The primary routes of exposure to these compounds are inhalation and skin absorption. The persistent nerve agent VX is one of the most highly toxic compounds following skin exposure (estimated human LD50 ∼10 mg). It has a low volatility, i.e. 8–10 mg m−3 at 25°C, and thus mainly presents a skin contact hazard. VX was shown to rapidly penetrate the skin both using in vivo and in vitro skin models. The clinical signs of nerve agent poisoning (resulting from inhibition of acetylcholinesterase) were demonstrated to rapidly appear in domestic swine challenged with VX, with death occurring within one to two hours.
The Effect of Sulfur Mustard Exposure and Freezing on Transdermal Penetration of Tritiated Water Through Ex Vivo Pig Skin
Authors: Payne O. J. , Graham S. J. , Dalton C. H. , Spencer P. M. , Mansson R. , Jenner J. , Azeke J. , Braue E.
Published: December 2013

Sulfur mustard (SM) is a well known and potent vesicating agent that has been used in many conflicts during the twentieth century. Cutaneous sulfur mustard injuries are often painfully debilitating, requiring surgical intervention (e.g., tissue debridement) and specialist burn care support. Wounds can take months to heal and place a substantial burden on the medical chain. Topical pharmaceutical formulations as medical countermeasures to cutaneous sulfur mustard injuries carry important advantages, including ease and non-invasiveness of dermal application, and effectiveness in reducing SM associated skin damage.
Modulation of Sulphur Mustard Skin Absorption by Topical Delivery of Fatty Acids and Cholesterol
Authors: Chilcott R. P. , Hutton O. J. , Jenner J.
Published: December 2013

Sulphur mustard (SM) is a chemical warfare agent that causes severe skin lesions. Previous work has shown that 80% of an applied liquid droplet of SM will vaporise from the skin surface before absorption can occur. There are a number of potential methods to increase the vapour loss of SM from the skin surface (thereby decreasing the dose received by the skin). These include use of an appropriate barrier cream or topical application of compounds that limit partitioning of SM into the skin.
Protective and Temporal Effects of Clothing on the In Vitro Absorption of Chemical Warfare Agents and Simulants
Authors: Matar H. , Price S. C. , Chilcott R. P.
Published: December 2013

The purpose of the study was to investigate the effect of common clothing fabrics on the percutaneous absorption of three chemical warfare agents (sulphur mustard (HD), soman (GD) and VX) and a chemical warfare agent simulant (methyl salicylate; MS).
Production of hydrogen by electrocatalysis: making the H–H bond by combining protons and hydrides
Authors: Bullock R. Morris , Appel Aaron M. , Helm Monte L.
Published: December 2013

Electrocatalytic production of hydrogen by nickel complexes is reviewed, with an emphasis on heterocoupling of protons and hydrides.
Rationale for the sluggish oxidative addition of aryl halides to Au(i)
Authors: Livendahl Madeleine , Goehry Charles , Maseras Feliu , Echavarren Antonio M.
Published: December 2013

Linear 2-coordinate d10 Au(i) complexes could form Au(iii) species by oxidative addition with reasonable energies, although the barriers to access them are prohibitively high.
Molecular dynamics approaches to the design and synthesis of PCB targeting molecularly imprinted polymers: interference to monomer–template interactions in imprinting of 1,2,3-trichlorobenzene
Authors: Cleland Dougal , Olsson Gustaf D. , Karlsson Björn C. G. , Nicholls Ian A. , McCluskey Adam
Published: December 2013

Molecular dynamic simulations identify MeOH as disrupting the FM–T interactions and reducing imprinting efficacy with 1,2,3-trichlorobenzene (2).
Spatially addressable design of gradient index structures through spatial light modulator based holographic lithography
Authors: Ohlinger Kris , Lutkenhaus Jeff , Arigong Bayaner , Zhang Hualiang , Lin Yuankun
Published: December 2013

In this paper, we present an achievable gradient refractive index in bi-continuous holographic structures that are formed through five-beam interference. We further present a theoretic approach for the realization of gradient index devices by engineering the phases of the interfering beams with a pixelated spatial light modulator. As an example, the design concept of a gradient index Luneburg lens is verified through full-wave electromagnetic simulations. These five beams with desired phases can be generated through programming gray level super-cells in a diffractive spatial light modulator. As a proof-of-concept, gradient index structures are demonstrated using synthesized and gradient phase patterns displayed in the spatial light modulator.
Deep subwavelength plasmonic waveguide switch in double graphene layer structure
Authors: Iizuka Hideo , Fan Shanhui
Published: December 2013

Graphene provides excellent prospects of the dynamic tunability, low propagation loss, and extreme mode confinement for plasmonic excitations in the infra-red and terahertz frequencies. We show that in a deep subwavelength double graphene layer structure, graphene plasmons can be routed between two different graphene waveguides by relatively small chemical potential tuning. We develop a coupled mode theory that completely accounts for the switching behavior observed in numerical simulations. Such a deep subwavelength 1 × 2 device is a crucial enabling component towards large-scale integrated deep-subwavelength electromagnetic circuits.
Anisotropy of effective electron masses in highly doped nonpolar GaN
Authors: Feneberg Martin , Lange Karsten , Lidig Christian , Wieneke Matthias , Witte Hartmut , Bläsing Jürgen , Dadgar Armin , Krost Alois , Goldhahn Rüdiger
Published: December 2013

The anisotropic effective electron masses in wurtzite GaN are determined by generalized infrared spectroscopic ellipsometry. Nonpolar (112¯0) oriented thin films allow accessing both effective masses, m⊥* and m∥*, by determining the screened plasma frequencies. A n-type doping range up to 1.7 × 1020 cm−3 is investigated. The effective mass ratio m⊥*/m∥* is obtained with highest accuracy and is found to be 1.11 independent on electron concentration up to 1.2 × 1020 cm−3. For higher electron concentrations, the conduction band non-parabolicity is mirrored in changes. Absolute values for effective electron masses depend on additional input of carrier concentrations determined by Hall effect measurements. We obtain m⊥*=(0.239±0.004)m0 and m∥*=(0.216±0.003)m0 for the parabolic range of the GaN conduction band. Our data are indication of a parabolic GaN conduction band up to an energy of approximately 400 meV above the conduction band minimum.
Ultrasound biomicroscopy: a non-invasive approach for in vivo evaluation of oocytes and small antral follicles in mammals
Authors: Pfeifer L. F. M. , Adams G. P. , Pierson R. A. , Singh J.
Published: December 2013

The use of ultrasonography has changed our understanding of the ovarian function in live animals. However, most of the studies that have used ultrasonography to image the ovary have provided data only of structures >1 mm in diameter. The recent availability of high-resolution ultrasound technology with high-frequency transducers (25–70 MHz), offers the potential to examine the developmental dynamics of small antral follicles and the cumulus–oocyte complex (COC) in vivo. In this review we provide data from a series of studies performed by Veterinary Biomedical Sciences Laboratory describing the advantages and disadvantages, as well as image characteristics, of ultrasound biomicroscopy (UBM) to study ovarian biology in mammals. Data and images of small ovarian structures in rabbits, cattle, mice and humans are shown. The UBM technique allowed visualisation of small antral follicles ranging in size from 300 to 700 μm in all species examined, as well as COC within follicles in rabbits, cattle and humans. Furthermore, UBM permitted clear distinction of the follicular wall from the surrounding ovarian stroma in cattle and humans. At present, the limited depth of penetration of UBM restricts the use of this technique to an experimental setting. In that regard, further studies using UBM will probably result in a greater understanding of the pattern and control of early antral folliculogenesis and oogenesis.
10 INFLUENCE OF ADMINISTRATION OF HUMAN CHORIONIC GONADOTROPIN ON DAY 4 AFTER FIXED-TIME AI ON CORPUS LUTEUM SIZE, FUNCTION, AND PREGNANCY RATE IN BEEF COWS
Authors: Borges J. B. S. , Thedy D. X.
Published: December 2013

Successful establishment of pregnancy in the cow depends on early rise of progesterone (P4) to improve embryo development. The present study was carried out to determine the corpus luteum (CL) function and pregnancy rate following the administration of 1.500 IU of hCG in suckled beef cows on Day 4 after fixed-time AI (FTAI). Crossbred Angus cows (n = 429), 40 to 70 days postpartum, were synchronized using 2 mg of oestradiol benzoate (EB) i.m. and an intravaginal progesterone-releasing device containing 1 g of P4 for 7 days. At device removal, prostaglandin F2a (150 mg of D-cloprostenol) was injected i.m.; 24 h later, 1 mg of EB was given and the FTAI were done 30 to 36 h after EB using semen of just 1 bull. Cows were assigned alternatively to receive 1.500 IU of hCG i.m. (n = 200) or 1 mL of saline i.m. (n = 229) on Day 4 after FTAI. Ovaries were examined by transrectal ultrasonography (n = 71) on Days 4 and 7 to detect the presence and to measure the area of CL. Blood samples were collected on Days 4, 7, 10, 14, and 30 post-oestrus to determine P4 concentration by RIA. Pregnancy diagnoses were done 30 days after FTAI. The effects of treatments on CL area and serum P4 levels were analysed by ANOVA (Proc GLM, SAS Institute Inc., Cary, NC, USA) and pregnancy rates compared by chi-square (Proc CATMOT, SAS). The induction of accessory CL determined on Day 7 was 44% in cows treated with hCG and not observed in the control group. The area of the CL on Day 7 was significantly higher (P < 0.01; 3.37 ± 2.56 v. 2.56 ± 0.22 cm2) in the hCG group. Despite the increase in CL size, the mean levels of P4 on Days 4, 7, 10, 14, and 30 post-AI indicated no difference (P > 0.05) in CL function between control and hCG-treated cows. On Day 30, pregnancy rates were not different (P = 0.08), at 46.7 and 55% for control and hCG-treated cows, respectively. In conclusion, administration of 1.500 IU of hCG on Day 4 post-FTAI did not improve pregnancy rate in suckled beef cows.
102 INCLUSION OF A RECOMBINANT BOVINE TRYPSIN INTO THE IETS SANITARY PROTOCOL FOR DISINFECTION OF BOVINE EMBRYOS TO PREVENT BOVINE HERPESVIRUS 1 (BHV 1) TRANSMISSION VIA EMBRYO TRANSFER TO RECIPIENTS AND CALVES
Authors: Bielanski A. , Algire J. , Lalonde A. , Garceac A.
Published: December 2013

This study dealt with the potential for introduction of infectious agents through the use of animal-derived products. The efficacy of a recombinant bovine trypsin (RBTr) as a replacement for porcine pancreatic trypsin as a disinfectant for bovine herpesvirus-1 (BHV-1)-infected embryos was investigated according to the sanitary guidelines of the IETS. Treatment of in vivo-fertilised embryos (n = 147) contaminated with BHV-1 [105 50% tissue culture infectious dose (TCID50)/mL] in the presence of RBTr (525 U mL–1) for 120 s effectively removed the infectious virus (100%) compared with untreated and washed embryos [9/55 (16%); P < 0.05]. Transfer of in vivo-fertilised and disinfected embryos to BHV-1 seronegative recipients (n = 24) resulted in 14 pregnancies and 11 calves born free of BHV-1. In contrast, transfer of unwashed/undisinfected embryos to 4 recipients resulted in seroconversion and no pregnancies at term. The same treatment applied to in vivo-fertilised contaminated embryos removed the infectious virus from all but 1 [1/100 (99%)] compared with untreated and unwashed embryos [16/28 (57%); P < 0.05]. It was concluded that the use of RBTr can be considered as an alternative method of rendering embryos free of bovine herpesvirus and thus reduce the potential risk of disease transmission to embryo recipients and offspring.
120 IDENTIFICATION OF THE RELATIVELY PREDOMINANT BUFFALO INTERFERON tau ISOFORM AND ITS EXPRESSION IN ESCHERICHIA COLI
Authors: Saugandhika S. , Malik H. N. , Singhal D. K. , Singhal R. , Dubey A. , Boateng S. , Singh A. K. , Kumar S. , Kaushik J. K. , Mohanty A. K. , Upadhayay R. C. , Malakar D.
Published: December 2013

The maternal pregnancy recognition factor interferon tau (IFN-τ) is expressed in multiple isoforms in all pecoran ruminant species. Interferon-τ, as the first pregnancy signaling molecule, performs a significant role in implantation as well as establishment of pregnancy. Due to low reproductive efficiency of buffalo compared with bovine and IFN-τ being the key molecule of reproductive physiology in ruminants, the objective of our study was framed to identify the various IFN-τ transcripts in buffalo embryonic trophoblast cells, to know their relative abundance to identify the relatively predominant isoform, and lastly to clone and express it in a heterologous host. Following total cellular RNA extraction from primary trophectodermal cells, RT-PCR was performed using gene-specific primers designed against known bovine IFN-τ sequence. Cloning of the amplified product and screening of the recombinant colonies gave 13 distinct cDNA variants that encoded for 8 distinct buffalo IFN-τ isoforms. These buffalo IFN-τ isoforms have a greater nucleotide and amino acid homology with caprine IFN-τ (98–100% and 96–100%) than ovine (94–97% and 90–95%) and bovine (89.6–90.6% and 82–86%), respectively. The novel buffalo IFN-τ isoforms showed pronounced nucleotide and amino acid sequence identity with one another (99.1–99.8% and 98–99%) but only moderate identity with previously identified buffalo IFN-τ (90–92% and 82–86%). All the 13 transcript sequences were accepted in GenBank. Out of 8 isoforms, buffalo IFN-τ1 has been found to be the relatively predominant, which was subcloned into expression vector pET 22b without signal sequence from pJET cloning vector and expressed in competent BL21 (DE3) Escherichia coli strain. Expression of the recombinant protein in soluble form was induced by isopropyl β-D-1-thiogalactopyranoside (0.1 mM) at 30°C for 6 h. The recombinant BuIFN-τ obtained was confirmed by Western blot using anti-HIS antibody. A new 20-kDa protein was detected coinciding the molecular weight of IFN-τ reported earlier in literature. In conclusion, the current study revealed that there are 8 different isoforms of IFN-τ that are expressed in trophectodermal out-growths during early pregnancy of buffalo. Predominantly found isoform IFN-τ 1 was expressed in pET 22b vector, and recombinant soluble protein was confirmed by Western blot.
125 BISPHENOL A-INDUCED GROWTH OF OVARIAN CANCER CELLS WAS REVERSED BY A PHYTOESTROGEN, GENISTEIN, BY INHIBITION OF A CROSSTALK BETWEEN ESTROGEN RECEPTOR AND INSULIN-LIKE GROWTH FACTOR 1 RECEPTOR IN IN VITRO AND XENOGRAFT MOUSE MODELS
Authors: Hwang K.-A. , Kim S.-H. , Choi K.-C.
Published: December 2013

It has been shown that oestrogen (E2) up-regulated the expression of components of insulin-like growth factor-1 (IGF-1) signaling pathway and induced the downstream of mitogenic signaling cascades via phosphorylation of insulin receptor substrate-1 (IRS-1). An interaction between oestrogen receptor (ER) and IGF-1 receptor (IGF-1R) signaling pathway plays an important role in proliferation of and resistance to endocrine therapy to oestrogen-dependent cancers (i.e. breast and endometrial cancers). In the present study, we evaluated xenoestrogenic effect of bisphenol A (BPA) and antiproliferative activity of genistein (GEN) in accordance with the influence on this crosstalk. The gene expressions in mRNA and protein levels were examined by semiquantitative RT-PCR and Western blot analysis, in which the primers for ERα, IGF-1R, and GAPDH and the antibodies against pIRS-1, pAkt, and GAPDH were used, respectively. Total RNA and protein samples were isolated from BG-1 cells treated with dimethyl sulfoxide (DMSO), estradiol (E2; 10–9 M), BPA (10–5 M), E2 (10–9 M) + GEN (10–4 M), and BPA (10–5 M) + GEN (10–4 M). The DMSO was used a vehicle of E2, BPA, and GEN in in vitro experiments. All in vitro experiments were done in triplicates. The effects on tumour growth and immunohistologic alterations were identified in in vivo mouse models. The mice were injected subcutaneously with corn oil (vehicle, n = 6), E2 (n = 6), BPA (n = 6), E2+GEN (n = 6), and BPA+GEN (n = 6) for 8 weeks. The BPA treatment resulted in up-regulation of ERα and IGF-1R mRNA, and induced phosphorylation of IRS-1 and Akt proteins compared with a control (DMSO) in BG-1 ovarian cancer cells as E2 did in triplicates. In the mouse model xenografted with BG-1 cells, BPA significantly increased a tumour burden of mice and expressions of ERα, pIRS-1, and cyclin D1 in tumour mass compared with the vehicle (corn oil), indicating that BPA induces ovarian cancer growth by promoting the crosstalk between ER and IGF-1R signals. On the other hand, GEN effectively reversed estrogenicity of BPA by reversing mRNA and protein expressions of ERα, IGF-1R, pIRS-1, and pAkt induced by BPA in cellular model with triplicates. The GEN also significantly decreased tumour growth and in vivo expressions of ERα, pIRS-1, and pAkt in a xenografted mouse model. Also, GEN was confirmed to have an antiproliferative effect by inducing apoptotic signaling cascades. Taken together, these results suggest that GEN effectively reversed the increased proliferation of BG-1 ovarian cancer by suppressing the crosstalk between ER and IGF-1R signaling pathways up-regulated by BPA or E2.
This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ009599), Rural Development Administration, Republic of Korea.
129 UTERINE EXPRESSION OF TRANSIENT RECEPTOR POTENTIAL MELASTATIN 2 CHANNEL AND ITS REGULATION BY SEX STEROID HORMONES
Authors: Ahn C. , Hong E. J. , Jeung E. B.
Published: December 2013

The transient potential receptor (TRP) channels are membrane-binding proteins that are non-selectively permeable for cations, such as Ca2+ and Mg2+, in numerous mammalian cells. The extracellular or intracellular ions play key roles in physiological function, including muscle contraction, cytokine production, insulin release, and apoptosis. Although TRPM channels have been implicated in the brain, bone marrow, and spleen, the presence of TRPM2 has been reported in the endometrium of the uterus. To determine whether expression of the TRPM2 gene in the uterus is due to gonadal steroid hormones or a hormone-independent effect, the uterine TRPM2 gene was monitored in mature rats during the oestrous cycle and in immature rats after treatment with gonadal steroid oestrogen (E2), progesterone (P4) with/without their antagonist, ICI 182,780, and RU486. Dramatic induction of the level of TRPM2 mRNA occurs at proestrus, followed by a drop to baseline levels at metestrus, and its level is restored at diestrus. Furthermore, the immune-reactive TRPM2 is observed in stromal cells of the myometrium and endometrium, and changes during the oestrus cycle. In addition, E2-induced TRPM2 is inhibited by co-treatment with P4. Taken together, these results imply that TRPM2 expression levels in the uterus are regulated by gonadal steroid hormones E2 and P4. Results of this study suggest possible involvement of TRPM2 in reproductive function during the oestrous cycle in female rats.
134 EFFECT OF REPLACING FETAL BOVINE SERUM BY DIFFERENT GROWTH FACTORS IN THE PRE-IMPLANTATION DEVELOPMENT OF BOVINE EMBRYOS GENERATED BY IN VITRO FERTILIZATION
Authors: Felmer R. , Vargas T. , Sanchez R. , Arias M. E.
Published: December 2013

Different culture systems have been studied that support pre-implantation development of bovine embryos up to the blastocyst stage. However, the use of chemically defined culture systems has been less studied. The objective of the present study was to evaluate the effect, in the developmental potential of in vitro-produced bovine embryos, of replacing fetal bovine serum (FBS) by different growth factors in the maturation and embryo culture media. In experiment 1, oocytes collected by aspiration of ovaries from a local slaughterhouse were matured in standard TCM-199 culture medium at 38.5°C, 5% CO2, and saturation humidity. The effect of insulin-like growth factor 1 (100 ng mL–1), epidermal growth factor (10 ng mL–1), and fibroblast growth factor 2 (500 ng mL–1) was evaluated at 24 h by the presence of a polar body after removal of cumulus-oocyte complexes. In experiment 2, oocytes matured in vitro in the presence of FBS were fertilized by co-incubation with commercial sperm (mL) for 18 h in standard fertilization medium (Fert-TALP). The presumptive zygotes were denuded and randomly allocated in a chemically defined culture medium based on KSOM supplemented with polyvinyl alcohol (PVA), fructose, and each of the growth factors listed previously. Undefined cultured medium was based on KSOM supplemented with 5% FBS. Embryos were cultured at 38.5°C in a mixture of gases and saturation humidity. Cleavage and blastocyst rates were recorded on Days 3 and 7, respectively. Analysis of variance was used to test for statistically significant differences between groups (P < 0.05) using Stat Graphics Plus 2 Software. In cases where statistically significant differences were observed, a multiple comparison test was run using Tukey's test. In experiment 1, a similar maturation rate was observed in all treatments relative to the undefined maturation medium (range = 88–91%). In experiment 2, no differences were observed in the cleavage (79, 87, 85, and 85%) and the blastocyst rates (24, 25, 26, and 30%) for the epidermal growth factor, insulin-like growth factor 1, fibroblast growth factor 2, and FBS treatments, respectively. In conclusion, we demonstrated that maturation of bovine oocytes can be achieved in chemically defined conditions by replacing FBS by each of the growth factors evaluated herein. Furthermore, chemically defined KSOM medium supplemented by any of these growth factors can generate a similar rate of blastocyst than the undefined medium containing FBS. Analyses are under way to evaluate the effect of completely defined culture conditions (maturation and embryo culture) on the pre-implantation development of embryos produced in the presence of these growth factors.
14 CONCEPTION RATE OF BUFFALO HEIFERS TREATED WITH OESTRADIOL CYPIONATE TO INDUCE OVULATION IN TIMED ARTIFICIAL INSEMINATION PROGRAMS
Authors: Souza D. G. , Martins A. , Oba E. , Camargos A. S. , Moreira E. P. , Gallego C.
Published: December 2013

Reducing animal handling and costs in timed artificial insemination (TAI) systems are of great interest in animal production, regardless of the species. Thus, this study was carried out to investigate the effects of administering oestradiol cypionate (EC) or hCG to induce ovulation in buffalo (Bubalus bubalis) in TAI programs. The 79 Bufalipso heifers housed at Institute of Animal Science, San José de las Lajas, Cuba, were, on average, 24 months of age with a body weight of 310 to 380 kg at the beginning of the experiment. The investigation was conducted out of the breeding season (25 June through 28 July 2012). Heifers were randomly assigned to 1 of 3 treatment groups; either hCG (control group) or EC1 and EC2, in which TAI was performed at either 48 or 56 h after the EC injection, respectively. On the first day of the treatment regime (Day 0), animals received oestradiol benzoate (2 mg i.m.; Sincrodiol®, Ourofino, Brazil), followed by the insertion of an intravaginal device containing 1 g of progesterone (Primer®, Tecnopec, São Paolo, Brazil). On Day 9, an intramuscular luteolytic dose of cloprostenol (500 μg; Ciosin®, Intervet, Brazil) and eCG (500 UI; Novormon®, Intervet) were given in all heifers along with the removal of the intravaginal device. On the same day, 0.5 mg of EC (ECP®, Pfizer, Brazil) was administered intramuscularly in the EC1and EC2 groups. Animals in the hCG group received hCG (1000 IU i.m.; Vetecor®, Hertape Calier, Spain) on Day 11 and were inseminated 14 h later. Pregnancy diagnosis was performed by ultrasound 35 days after TAI. Data were analysed by a chi-squared test, with P < 0.05 taken as significant. There were no differences among treatment groups, with values of 57.7 (15/26), 60.7 (17/28), and 64% (16/25) for the EC1, EC2, and hCG groups, respectively. In conclusion, oestradiol cypionate was effective at inducing ovulation in buffalo heifers in TAI programs comparable to the control group, as evidenced by the conception rates. In addition, animals subjected to the EC protocol pass just 3 times through the corral instead of 4 when receiving hCG, which decreases the animal handling and costs.
153 ZINC INSUFFICIENCY DURING PORCINE OOCYTES IN VITRO MATURATION CAUSED MEIOTIC BLOCK AND DEVELOPMENTAL FAILURE
Authors: Kim E. , Jeon Y. , Yoon J. D. , Cai L. , Hwang S. U. , Lee S. , Hyun S. H.
Published: December 2013

The objective was to investigate the effects of zinc (Zn) insufficiency during in vitro maturation (IVM) of porcine oocytes. Zinc insufficiency was induced by treatment of Zn chelator, N,N,N′,N′-tetrakis-(2-pyridylmethyl)-ethylendiamine (TPEN). In experiment 1, we investigated the effect of duration of Zn insufficiency in IVM on oocytes maturation and subsequent embryonic development after parthenogenetic activation (PA). First, 10 μM TPEN was added to the IVM medium for 0, 7, 15, or 22 h. After TPEN treatment, 10 μM Zn were supplemented on IVM medium except in the 0 h group. Reductions in the nuclear maturation rates were dependent on TPEN duration. The 0-h-treated oocytes showed 83.9 ± 3.9% metaphase II (MII) rate; the 7-h-treated oocytes had significantly lower MII rate (44.8 ± 3.0%) than 0-h-treated oocytes. The majority of 15- and 22-h-treated oocytes were arrested at metaphase I (MI rate: 98.0 ± 1.0 and 97.2 ± 1.7%, MII rate: 0 and 0%, respectively). Embryonic developmental competence was similar to maturation results. Reduction in cleavage and blastocyst (BL) rates were also dependent on duration of TPEN treatment (cleavage rate: 65.3 ± 1.4, 42.6 ± 4.8, 2.6 ± 0.1, and 3.0 ± 1.6%; BL formation rate: 29.3 ± 2.8, 9.2 ± 1.5, 0, and 0% for 0, 7, 15, and 22 h). Total cell number of BL was also significantly different. Total cell number of BL in the 0-h-treated group (51.4 ± 4.5) was significantly higher than that in the 7-h-treated group (23.2 ± 1.6). In experiment 2, to confirm that the Zn insufficiency caused oocyte immaturities and loss of developmental competence in TPEN-treated oocytes, we investigated nuclear maturation and subsequent embryonic development following 3 groups: (1) non treatment (control); (2) 10 μM TPEN treatment during 22 h of IVM; (3) 10 μM TPEN + 10 μM Zn treatment during 22 h of IVM. Only TPEN-treated oocytes and TPEN+Zn-treated oocytes showed contrasting results. Oocyte maturation rates and subsequent embryonic development competence in TPEN with Zn-treated oocytes were similar to control (MII rate: 93.0 ± 1.2 and 92.7 ± 1.8%, BL formation rate: 42.0 ± 6.7 and 40.0 ± 7.5% for TPEN+Zn-treated oocytes and control). These results were significantly different compared with only TPEN-treated oocytes’ results (MII rate: 0.61 ± 0.61%, BL formation rate: 0%). In conclusion, Zn is an essential element for successful oocyte maturation and embryo development in porcine. Zinc insufficiency caused meiotic block and had lasting effects on early embryo development.
This work was supported, in part, by a grant from the Next-Generation BioGreen 21 Program (No. PJ00956901), Rural Development Administration, and the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2012R1A1A4A01004885, NRF-2013R1A2A2A04008751), Republic of Korea.