bluuebunny/crossref_metadata_2025 huggingface dataset:
Floral morphology and pollination system in the native Australian perennial pasture legume Cullen australasicum (syn. Psoralea australasica)
Authors: Wang Yan-Jing , Nair Ramakrishnan M. , Mu Chun-Sheng , Dundas Ian S.
Published: August 2011

Cullen australasicum (syn. Psoralea australasica) is a native perennial legume with potential in the low-rainfall wheatbelt of southern Australia. The objective of this study was to investigate the reproductive biology of C. australasicum utilising five accessions. Glasshouse and field pollination experiments were conducted in Adelaide, South Australia. Floral morphology, stigma receptivity and pollen : ovule ratios were determined. Pollen tube growth and stigma morphology were examined using fluorescence and scanning electron microscopes. Glasshouse pollination studies indicated that four of the accessions showed the need for an external tripping agent to bring about pollination and that hand-tripping was the most efficient method. A scanning electron microscopy study revealed there are two types of stigmas in this species. Stigma receptivity was significantly lower at the early bud stage before anther dehiscence. The results show that C. australasicum is a self-compatible species comprising accessions with a wide range of outcrossing potential.
Phosphorus-efficient faba bean (Vicia faba L.) genotypes enhance subsequent wheat crop growth in an acid and an alkaline soil
Authors: Rose Terry J. , Damon Paul , Rengel Zed
Published: August 2011

Faba bean (Vicia faba L.) is a carboxylate-exuding legume that enhances the phosphorus (P) nutrition of subsequently grown cereals. In an earlier study we found variation in soil P acquisition among 50 faba bean genotypes, but little is known about the rhizosphere processes that may contribute to P efficiency and whether these processes impact on the growth of subsequent cereal crops. In this study, we investigated rhizosphere dynamics (P fractions depleted, pH and carboxylate exudation) in three P-inefficient and five P-efficient faba bean genotypes in a glasshouse study on soils differing in P dynamics. The results suggest that P efficiency in the acidic soil was not driven by rhizosphere processes, consistent with earlier findings that root growth parameters contributed to P efficiency in this soil. In contrast, in the alkaline soil the most P-efficient genotypes had the highest malate exudation, which might enhance P solubilisation. For the first time, we showed a faba bean genotype-specific enhancement of growth and P uptake of subsequently grown wheat plants. This genotypic variation could be exploited to further increase the benefit of faba beans in rotation with wheat on P-limited soils.
Arsenic speciation in bodily fluids of harbor seals (Phoca vitulina) and harbor porpoises (Phocoena phocoena)
Authors: Kuenstl Linda , Griesel Simone , Prange Andreas , Goessler Walter
Published: August 2011

Environmental context. Marine mammals play an important role in their ecosystem. As top predators they ingest a lot of arsenic from their food. In the present study bodily fluids and tissue samples of harbor seals and porpoises were investigated for arsenic speciation in order to obtain a clearer picture on their feeding habits and consequently a better understanding of the mass mortality of the animals in the North and Baltic Sea. Abstract. The total arsenic concentrations and arsenic speciation in various tissues and bodily fluids of harbor seals (Phoca vitulina) and harbor porpoises (Phocoena phocoena) were determined to obtain information about the feeding habits of these endangered marine mammals. Lower whole blood arsenic concentrations were found for fish-fed (median: 71 μg As L–1) than for free ranging seals (median: 190 μg As L–1). In porpoise liver the arsenic concentrations were higher from carcasses found in the North Sea (median: 421 μg As kg–1 wet mass) than from those inhabiting the Baltic Sea or found in the River Elbe (median: 250 μg As kg–1). The arsenic speciation in the urine, plasma, and gastric juice of seals and the urine of porpoises, collected from animals at different areas in the North and Baltic Seas, revealed the following picture: arsenobetaine was the predominant arsenic species in all measured bodily fluids. Plasma samples of seals contained only dimethylarsinic acid as additional species. In gastric juice arsenocholine and trimethylarsine oxide were found at trace concentrations. Several arsenic compounds were identified in mammals’ urine, the major being dimethylarsinic acid and thio-dimethylarsinic acid but high variability was observed in the relative proportions of each. No correlation between degree of decay and arsenic speciation in urine could be found. This is very useful information as older carcasses can also be included in future studies. Our preliminary results are promising to obtain an insight into feeding habits of seals and porpoises by the arsenic speciation in urine.
Hexabromocyclododecanes in human adipose tissue from Japan
Authors: Isobe Tomohiko , Oda Hiroko , Takayanagi Nozomi , Kunisue Tatsuya , Komori Hiroaki , Arita Norimasa , Ueda Norifumi , Nose Masato , Yamada Taketo , Takahashi Shin , Tanabe Shinsuke
Published: August 2011

Environmental context. Contamination by hexabromocyclododecanes (HBCDs), a group of brominated flame retardants, is of great concern due to their bioaccumulative nature and toxic implications. HBCD consumption in Japan is the highest among Asian countries and is still growing. In this regard, human exposure to HBCDs is of great concern in Japan. We present here the results of our investigation on Japanese human adipose tissue using LC-MS/MS. HBCDs were detected in all the samples analysed, and were generally higher in men than in women.
Abstract. Residue levels of hexabromocyclododecanes (HBCDs) were measured using LC-MS/MS in the Japanese human adipose tissue samples collected during 2003–2004 from Ehime prefecture (n = 26) and Kanto region (n = 9) of Japan. Concentrations of HBCDs (0.85–39 ng g–1 lipid) in the adipose tissue samples were 1–2 orders of magnitude lower than those of organochlorines (OCs). Regional differences between HBCD levels in Ehime prefecture and Kanto region were not significant. No age-dependent accumulation of HBCDs was observed. HBCD levels in men were significantly higher than those in women. Significant positive correlation between polybrominated diphenyl ethers (PBDEs) and HBCDs implies similar routes of exposure to these contaminants for Japanese citizens. α-HBCD was predominant among the three isomers, which is consistent with the other reports on bioaccumulation of α-HBCD in higher trophic animals.
Spatial and temporal variation in the structure of estuarine macroinvertebrate assemblages: implications for assessing the health of estuaries
Authors: Hirst A. J. , Kilpatrick R.
Published: August 2011

As human impacts in estuaries are often pervasive (estuary-wide) and/or pre-existing, the identification of suitable reference points, from which to assess the extent of impacts, is problematic. One solution is to compare potentially degraded estuaries with estuaries deemed to be largely unmodified by human activities. However, there is a perception that individual estuaries are too spatially and temporally dynamic to allow valid comparisons to be made using such an approach. We tested this idea for a commonly used indicator, benthic macroinvertebrates, using a factorial design incorporating both temporal and spatial scales between and within three adjacent meso-tidal river estuaries in northern Tasmania. Variation in macroinvertebrate assemblage structure was analysed using permutational multivariate analysis of variance. Most variance occurred within estuaries (68–82% variance) relative to spatial differences between estuaries (24–14%) corresponding with a strong upstream estuarine gradient and small-scale spatial patchiness. Seasonal variation accounted for 9–4% of total variance indicating that temporal differences were relatively insignificant when contrasted against spatial variability within and between estuaries. We suggest that with sufficient spatial replication at the within estuary-scale, entire estuaries may act as whole reference systems, allowing studies to examine potential impacts within estuaries with spatially diffuse, pre-existing human impacts.
Variable growth band deposition leads to age and growth uncertainty in the western wobbegong shark, Orectolobus hutchinsi
Authors: Chidlow Justin A. , Simpfendorfer Colin A. , Russ Garry R.
Published: August 2011

Age and growth parameters of Orectolobus hutchinsi were estimated using micro-radiographs of sectioned vertebrae from 182 wild caught individuals. Two fluorochrome marker dyes, calcein and oxytetracycline, were used to validate the timing and periodicity of vertebral band formation in nine individuals held in the laboratory for between 423 and 472 days. Growth bands were difficult to interpret and final counts were obtained from only 98 (53.8%) individuals ranging in total length (TL) from 63 to 146 cm. The timing of growth band formation in the vertebrae of captive animals had no predictable temporal pattern, with formation occurring during all seasons of the year, making age validation difficult. Growth band formation was hypothesised to be influenced by non-periodic changes in centrum or somatic growth rather than seasonal growth, as observed in many other elasmobranch species. Growth rates of nine O. hutchinsi held in captivity varied considerably, ranging from 3.5 cm year–1 to 13.8 cm year–1 in total length (mean = 7.03 cm year–1). Although the periodicity of vertebral band formation in captive animals did not support a synchronous annual pattern, captive growth rates matched those predicted when an annual band pattern was assumed for wild caught individuals. Von Bertalanffy growth parameters estimated from vertebral analysis assuming an annual banding pattern and a mean size of birth of 24.1 cm were: L∞ = 149.45 cm and K = 0.117 year–1 for both sexes combined. These results illustrate the fundamental importance of validating the periodicity of growth band formation in elasmobranch age and growth studies as it has considerable implications for the management of fisheries that exploit shark and ray species that may exhibit asynchronous growth band deposition.
Do bacteria regenerate phosphorus while decomposing seston?
Authors: García-Ruiz R. , Lucena J. , Niell F. X.
Published: August 2011

The C:P ratio of both seston in suspension and seston collected by sediment
traps were systematically higher than the Redfield ratio of 106 : 1 (molar)
and decreased significantly with depth in the mesotrophic, phosphorus-limited
La Concepción reservoir. An aerobic decomposition experiment of seston
at different concentrations by a mixed population of reservoir bacteria was
performed in batch cultures to investigate the bacterial role in the changes
of the C:P ratio. Dissolved phosphorus was taken up during the bacterial
growth phase, while carbon was transformed to CO2 .
Therefore, the C:P ratio of the decomposing seston decreased during this
stage, and could partially explain the vertical distribution for the C:P ratio
in this reservoir. During the death phase of the bacteria, flagellate grazing
caused most of the bacterial phosphorus to be released to the medium.
Seasonal variation of surface water CO2 partial pressure in the Southland Current, east of New Zealand
Authors: Currie Kim I. , Hunter Keith A.
Published: August 2011

The partial pressure of CO2 in the surface waters
(pCO2sw) of
the Southland Front east of New Zealand was determined over one annual cycle
in 1992/93. Associated parameters of temperature, salinity and nutrients
were also measured. Both the modified subtropical waters of the Southland
Current and the cooler subantarctic surface waters were undersaturated with
CO2 with respect to the atmosphere. The maximum
ΔpCO2 of –150
µatm was measured in April 1993. A simple model taking into account the
effects of temperature, biological activity and air–sea exchange applied
to subantarctic surface water indicated that at different times of the year
changes in sea surface temperature, biological activity and mixing processes
are the dominant controls of
pCO2sw .
Getting a foot in the door: can expanding the role of podiatry assistant improve access to public podiatry services?
Authors: Bergin Shan
Published: August 2011

The aim of this pilot was to evaluate the impact delegation of identified tasks to a qualified podiatry assistant (PA) had on accessibility to footcare services for targeted client groups. The role of PA was defined for the purposes of the pilot. The PA underwent theoretical and practical training and the role was implemented within a community based podiatry unit. Outcome measures included: total number of clients seen, change in available (qualified) podiatry hours through delegation of tasks to the assistant, changes to waiting times for initial and follow-up appointments, adverse events and patient satisfaction. The PA provided footcare to 41 clients during the evaluation period (December 2005 to January 2006 inclusive). Available (qualified) podiatry hours increased by 10 h per week for the same period and waiting times for an initial appointment were reduced from 12 to 2 weeks for the target group and from an ‘indefinite’ waiting period to 12 weeks for follow-up appointments. No adverse events were reported during the pilot. The findings of this pilot have implications for improving affordability and accessibility of footcare services to a significant proportion of the community and reducing the burden on current community based services.
Doctor supply in Australia: rural - urban imbalances and regulated supply
Authors: Kamalakanthan Abhaya , Jackson Sukhan
Published: August 2011

We review the debate on the supply of doctors in Australia from an economic perspective. We focus on the supply between urban and rural areas and on Australia’s reliance on foreign-born overseas-trained doctors. Documented evidence shows that doctors are concentrated in cities and rural Australians have relatively poor access; and there is heavy reliance on the recruitment of foreign doctors. We suggest that besides training more local doctors, policy-making should include innovations to resolve the supply imbalance such as physician assistants and community pharmacy care in areas where access to general practitioners is often limited.
Capitalising on the social resources within culturally and linguistically diverse communities for mental health promotion: stories of Australian Chinese people
Authors: Chan Bibiana
Published: August 2011

Social capital, generally observed in Chinese cultures, can be considered as the glue that holds together all the norms, trust and social networks that enhance mutual benefit of a collectivistic society. The aim of this paper is to explore the best way to tap into these social resources in mental health promotion. A mixed-method study, consisting of 16 community focus groups and a quantitative survey (n = 528), was conducted among Chinese Australians (recruited from general practices) in metropolitan Sydney. Although the focus groups explored help-seeking behaviours during emotional distress, the survey assessed the prevalence of such episodes. Chinese informants identified ‘family and friends’, ‘cultural values’, ‘spirituality’ and ‘self’ as common ‘helpers’ for managing depressive episodes before professionals were consulted. These ‘helpers’ function as the ‘social capital’ within the Chinese community. Focus group narratives of the low-acculturated Chinese revealed that they turned to close friends and family for help during crises. Highly acculturated Chinese found ‘family values’ most helpful when facing adversity. Survey findings indicated that the self-report rate of lifetime depression in low-acculturated Chinese was significantly lower than that of Australians. These results resonate with Cullen and Whiteford’s (2001) proposition that a higher level of social capital decreases the incidence of depression. Although the links between social capital and mental health require further investigation, the current findings support its role in counteracting emotional distress. There are implications of the current research findings to wider culturally and linguistically diverse communities.
114 BOVINE EMBRYO-MATERNAL RECOGNITION MAY OCCUR EARLIER IN AI THAN IN EMBRYO TRANSFER (ET)
Authors: Ideta A. , Nakamura Y. , Tsuchiya K. , Fujiwara H. , Yamaguchi T. , Imakawa K. , Aoyagi Y.
Published: August 2011

Embryo-maternal recognition is known to be mediated by the trophoblast and/or its secretory product during the preimplantation period. However, it was recently proposed that embryo-maternal recognition (along with the maternal immune system) could be activated by zona pellucida-degradation products including oligosaccharide chains from very early stage embryos (Fujiwara et al. 2009 J. Reprod. Immunol. 81, 1–8). If this is true, we predict that mammalian mothers recognise the presence of an embryo immediately after fertilization in vivo. The purpose of the present study was to compare the expressions of immune tolerance genes in bovine endometrial-luminal lymphoid (ELL) cells collected from animals after AI or embryo transfer (ET). ELL cells were collected from recipient cows inseminated with frozen–thawed semen on Day 0 (=oestrus, n = 8) and from cows that received a transferred embryo on Day 7 (n = 8) of the oestrous cycle. On Days 9 and 11, ELL cells ipsilateral to the corpus luteum were flushed non-surgically from four cows of each group using Ringer's solution (500 mL). To eliminate large cell clusters of endometrium and the embryo (blastocyst), the flushing medium was passed through an embryo collection filter (EmCon filter, 75 μm stainless steel screen). A single viable blastocyst was collected from each recipient cow. The recovered flushing medium containing ELL cells was centrifuged. The ELL cells were suspended in PBS and separated by Lympholyte-H (Cedarlane, Ontario, Canada) gradient centrifugation. Using RT-real-time PCR, transcripts for four immune tolerance genes in ELL cells from AI or ET examined were transforming growth factor-β (TGF-β), macrophage-colony stimulating factor (M-CSF), interleukin-10 (IL-10), and forkhead box P3 (Foxp3). Data were normalized to that of an endogenous control (glyceraldehyde-3-phosphate dehydrogenase) and analysed using ANOVA followed by Scheffe′s test. Transcripts for TGF-β, M-CSF, IL-10, and Foxp3 were detected in all RT-samples derived from ELL cells on Days 9 and 11 of the oestrous cycle. Transcript levels of TGF-β, M-CSF and IL-10 in the AI group were the same as those of the ET group (P > 0.05), although the transcript level of Foxp3, a master regulator of the function of regulatory T-cells, in the AI group was significantly higher than that of the ET group on Days 9 and 11 (P < 0.05). Furthermore, the transcript level of Foxp3 in the ET group on Day 11 was significantly higher than that of the ET group on Day 9 (P < 0.05). These results indicate that the embryo-maternal recognition in AI occurs earlier than in ET, and suggest that cows recognise an embryo earlier than those generally believed.
144 HEAT TOLERANCE OF DEMI-EMBRYOS FROM A TROPICAL-ADAPTED BOS TAURUS BREED
Authors: Alvarez R. H. , Martinez A. C. , Pires R. M. L.
Published: August 2011

Long-time selection of autochthonous Bos taurus breeds in a tropical environment is known to result in some physiological changes, including gamete thermotolerance. The aim of this study was to evaluate the resistance to heat shock (HS) of demi-embryos from tropical-adapted B. taurus cattle. The experiment was carried out in a tropical wet climate, at the experimental station of Instituto de Zootecnia (latitude 22°46′S; longitude 47°17′W) from November to February (average maximum temperature = 30.0 ± 0.8°C, and average absolute precipitation = 153.1 ± 78.8 mm3). Embryos (morulae) from 37 superovulated Caracu (a local B. taurus breed) cows were collected by uterine flushing on Day 6 after AI. Sixty-two grade I morulae were split with a metal microblade coupled to a micromanipulator, and each demi-embryo was placed separately in a 35-μL drop of culture medium [SOF supplemented with modified Eagle medium amino acids, sodium citrate, myo-inositol (SOFaaci), and 10% FCS] under mineral oil in a humidified atmosphere of 5% CO2, at 38.5°C for 24 h (control) or at 41°C for 12 h and a further 12 h at 38.5°C (HS). Halves of morulae that developed to the blastocyst stage (showing a central cavity) after a 24-h culture were transferred to recipient heifers previously synchronized with the donors. Pregnancy was monitored by ultrasound scanning 40 days after transfer, and data were analysed by chi-square test. There was a significant difference in the development to the blastocyst stage between control (44/62, 71.0%) and HS (30/62, 48.4%) demi-embryos (P < 0.01). Pregnancy rate obtained from control (14/44, 31.8%) and HS (8/30, 26.7%) blastocysts was not different (P > 0.05). We conclude that HS affected embryonic development from the morulae to blastocyst stage of cultivated demi-embryos, although fertility of blastocysts that survived to HS was not further compromised. Further studies are necessary to elucidate whether Caracu embryos are more thermotolerant than other tropically sensitive B. taurus breeds.

Supported by FAPESP.
149 AN INEXPENSIVE LABORATORY PRACTICE TO TEACH EMBRYO COLLECTION AND TRANSFER
Authors: Martinez A. , Tovar G. , Estrada C. , Esperon E. , Romo S.
Published: August 2011

The objective of this work is to describe a hands-on laboratory session for surgical embryo collection and transfer using rabbits as a model. Rabbits are characterised for their low purchase price, easy handling, and low maintenance, thus allowing their use as a tool for embryo transfer teaching and for its potential application in other species, such as the bovine, ovine, and caprine. These are important reasons in countries where the use of low-cost techniques and animal models is a must. Two females of the New Zealand breed were used as donors and one female of the Chinchilla breed was used as recipient. All females were healthy and multiparous. Three days after receiving natural service by a male, the donors were anesthetized (by injection of xylazine hydrochloride, 1.1 mg kg–1 IM, then sodium pentobarbital, 30 mg kg–1 IV), and ovariohysterectomy surgery was performed on each one. Each uterine horn was flushed 3 times with Ringer’s lactate (Hartmann) solution. Using a stereoscopic microscope, we searched for embryos and then placed them in a Petri dish for evaluation and selection. Once selected, embryos were moved to a 6-well dish containing 0.5 mL of a commercially available holding solution and washed 3 times. Twenty-one grades 1, 2, and 3 embryos were recovered and evaluated, showing developmental stages 1 to 6, according to IETS. The recipient female was anesthetized, and by laparotomy, her uterine horns were exteriorized to transfer a total of 15 grade 1 embryos with development stages 4 to 6. Pregnancy diagnosis was performed by real-time ultrasound on Days 10 and 20 post-transfer, with positive results. The transferred embryos induced a 34-day pregnancy, including 3 days of embryo development within the donors. Parturition was normal and without problems; 2 live offspring were born, with normal weight and size. The technique, used for teaching purposes, was successfully performed by 3 graduate students directed by 2 professors. The use of rabbits as a teaching model was efficient and the total cost of the laboratory practice was low, as compared with the use of other domestic species. The handling of the females and their offspring was easy, the feeding costs were low, and the practical laboratory session was fulfilled. It is proposed that the rabbit species be used for teaching surgical embryo collection and transfer to graduate and undergraduate students in underdeveloped and developing countries, where low-cost teaching models are of the utmost importance.
19 FACTORS AFFECTING PREGNANCY RATE AFTER ARTIFICIAL INSEMINATION WITH EQUINE SEMEN: PRODUCTION OF THE FIRST FOAL USING FROZEN SEMEN IN KOREA
Authors: Park Y. S. , Cho Y. G. , Cho K. H. , Cho G. J.
Published: August 2011

Research in the area of equine artificial insemination (AI) has led to its increased application in field trials (Loomis and Graham, 2008). However, procedures for equine semen collection, cooling and freezing of semen and artificial insemination need further improvement. This study examined the effect of semen preparation and ovulation on equine AI success. Stallion semen was collected via artificial vagina, diluted with skim-milk extender, and preserved by either cooling to 4°C or cryopreservation and storage in liquid nitrogen. Mares were examined by ultrasound for follicle development with every 12 h during oestrus. During AI, semen was inserted into uterine horn adjacent to the ovulating follicle sized over 4 cm and irregular shape. In experiment 1, mares were inseminated with either cooled, cooled-transported, or frozen–thawed semen. Cooled semen was used immediately after collection and cooling. Cooled-transported semen was used 4 h after semen collection and dilution. Semen freezing procedure was followed by Moore et al. (2006). In experiment 2, we examined the effect of buffer (skim-milk extender), which was infused into the uterus at the time of AI with frozen–thawed semen. In experiment 3, we compared AI pregnancy rates for mares ovulating spontaneously v. after treatment with hCG. Pregnancy data were analysed Chi square test where P-values < 0.05 were considered significantly different. In experiment 1, the pregnancy rates after AI using cooled, cooled-transported, and frozen–thawed semen were 60 (10/6), 50 (8/4), and 37.5% (8/3), respectively, and similar among treatments. In experiment 2, the pregnancy rate of mares infused with buffer at AI was 37.5% (8/3) which was higher than that with no buffer (0%; 6/0). In experiment 3, the pregnancy rates of mares were similar between ovulated spontaneously (25%; 8/2) and ovulated with hCG (50%; 8/4). The results suggest that equine semen that has been cooled, cooled and transported or frozen can be successfully used to establish AI pregnancy. Also, the pregnancy rates after AI can be increased by infusing buffer into the uterus at AI or by inducing ovulation with hCG. These experiments resulted in the first foal produced in Korea with frozen–thawed equine semen.

This research was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.
196 GLUTATHIONE S-TRANSFERASE THETA1 (GSTT1) IS DIFFERENTIALLY REGULATED FROM OTHER GLUTATHIONE S-TRANSFERASES IN GRANULOSA CELLS
Authors: Muraki M. , Takahashi Y. , Ishii T. , Kyuwa S. , Yoshikawa Y.
Published: August 2011

Oxidative stress is a major source of aging that damages genomic and mitochondrial DNA, causing female reproductive disorders. Glutathione S-transferases (GST) are known to detoxify the metabolites of genotoxic molecules to more water-soluble and readily excretable forms. Therefore, most GST have been considered to be down-regulated by aging. We recently demonstrated that only GSTT1 is up-regulated in granulosa cells from aged, infertile patients and aged mice, although other kinds of GST are down-regulated in these cells (Ito et al. 2008 Fertil. Steril. 90, 1026–1035). Measurement of the GSTT1 level in granulosa cells binding to the patient’s oocytes could be used in the selection of oocytes of high quality, meaning that the GSTT1 level in granulosa cells could be a good indicator of age-related infertility. To do that, an understanding of the concrete relationship between aging and GSTT1 up-regulation is necessary. However, the regulation mechanism of GSTT1 in granulosa cells remains unclear. In the present study, we attempted to examine the difference in expression mechanisms between GSTT1 and other GST. First, the expression patterns of GSTT1 and GSTP (a major member of the GST) in aged granulosa cells from patients were observed by immunostaining. The GSTT1 was up-regulated (young: 25 to 34 years old, N = 9; aged: 38 to 43 years old, N = 12; P < 0.05) but the GSTP was down-regulated (young: 28 to 30 years, N = 5; aged: 38 to 42 years, N = 7; P < 0.05) in the aged group. Nuclear factor erythroid 2 p45-related factor 2 (Nrf2) is a transcription factor that regulates the expression of most GST, so in the next experiment, we examined the expression patterns of GSTT1 and GSTP in granulosa cells from Nrf2-disrupted (Nrf2–/–) mice by immunostaining. The GSTP was down-regulated in granulosa cells of Nrf2–/– mice compared with those of Nrf2+/– mice, although a significant difference was not observed in GSTT1. In addition, we confirmed these results by using mouse embryonic fibroblasts (MEF) from Nrf2–/– and Nrf2+/– mice, and the same results were obtained. Our previous experiment showed that GSTT1 was induced by FSH in human granulosa-like tumour cells (KGN cells). We analysed the expression level of other genes in FSH-stimulated KGN cells. The transcription levels of GSTP and Nrf2 were not induced by FSH. These results suggest that the expression of GSTT1 is not regulated by Nrf2 and is differentially regulated from other GST. The difference in regulation mechanisms between GSTT1 and other GST may result in the up-regulation of GSTT1 in the aging granulosa cells, and it may contribute to the constitution of the diagnosis of oocyte quality using the GSTT1 measurement.
300 ESTABLISHMENT AND CHARACTERIZATION OF RAT MESENCHYMAL STEM CELLS
Authors: Kim T. H. , Jeon B. G. , Lee S. L. , Rho G. J.
Published: August 2011

Mesenchymal stem cells (MSC) are regarded as an attractive source for tissue engineering and regeneration, and bone marrow extract has been commonly used as a source of pluripotent MSC. However, skin tissue has recently been identified as a convenient alternative source of MSC. The present study was focused on the effect of characterised MSC derived from rat on expression of early transcriptional factors, alkaline phosphate (AP) activity, and in vitro differentiation into selected cell lineages. The MSC were isolated from 8-week-old s.d. rat’s ear skin and cultured in advanced DMEM supplemented with 10% fetal bovine serum at 37°C in a humidified atmosphere of 5% CO2 in air. To evaluate AP activity, cells were fixed with 3.7% formaldehyde solution and stained with Western Blue® (Promega, Madison, WI, USA). Expressions of early transcriptional factors (Oct-4, Sox2, and Nanog) were evaluated by RT-PCR. Differentiation into distinct mesenchymal lineages such as adipogenic, osteogenic, and neuron was done by following previously described protocols and assessed by lineage-specific stains. The specific genes in the osteocytes (osteocalcin, osteonectin, osteopontin, and Runx2), adipocytes (pparγ2, adiponectin, and aP2) or neuron (nestin, neurogenin 1, β-tublin, and nerve growth factor) were characterised by RT-PCR. The MSC were positive for AP activity and expressed Oct-4, Sox2, and Nanog. Following induction, MSC were successfully differentiated into adipocytes, osteocytes, and neurons. As adipocytes markers, aP2, pparγ2, and adiponectin were strongly detected in the adipocyte induced cells. Osteonectin, osteocalcin, Runx2, and osteopontin were expressed in the adipocyte induced cells. Futhermore, neuron-specific markers were clearly expressed in the neuronal differentiated cells. In conclusion, MSC have the capability of differentiation into multilineages including adipocytes, osteocytes, and neurons under the specific induction conditions. Skin tissue in rat can serve as an easily accessible and expandable alternative source for MSC harvesting and preclinical applications using an animal model.

This work was supported by Grant No. 2007031034040 from Bio-organ and 200908FHT010204005 from Biogreen21, Republic of Korea.
55 CONSTRUCTION AND IN VITRO DEVELOPMENT OF FOLLISTATIN TRANSGENIC PORCINE EMBRYO BY SOMATIC CELL NUCLEAR TRANSFER
Authors: Mu Y. S. , Jiang D. D. , Huan Y. J. , Zhu J. , Li H. , Liu Z. H.
Published: August 2011

Data from mice showed that follistatin can block myostatin activity in vivo and follistatin overexpressed in transgenic mice can induce dramatic increases in muscle mass. In order to determine whether this effect of follistatin have the same results in swine, we design experiment to make a follistatin transgenic porcine by somatic cell nuclear transfer. We hope to get a follistatin transgenic porcine that has double muscle mass. The present study aimed to construct follistatin transgenic embryos by somatic cell nuclear transfer. A follistatin (FST) expression vector, pFST-1, which contains a porcine FST cDNA (with modification of codon usage) driven by the human cytomegalovirus (CMV) promoter, was constructed. A SV40-neo expression cassette as a selection marker was inserted into pFST-1 to generate pST101, which was transfected into early-passage male primary porcine fetal fibroblast cells by the lipofection method; the transfected cells were selected with 400 mg mL–1 G418. The G418-resistant colonies were pooled. Cell genome DNA was extracted and PCR analysis showed that CMV promoter, follistatin CDS, and SV40 polyA can be detected, indicating that the expression construct had integrated in cell genome. RT-PCR analysis showed that pST101 cells contained higher amounts of FST cDNA compared with the non-transfected cells, indicating that the follistatin was overexpressed in the primary porcine cells. Then, pST101 cell as donor cell and non-transfected cells as control donor cell were used to construct somatic cell nuclear transfer (SCNT) embryos. All of these SCNT embryos were cultured in Porcine Zygote Medium-3. The cleavage rate and blastocyst rate was assessed on 48 h and Day 6. All data were subjected to a Generalized Linear Model Procedure (PROC-GLM) of Statistical Analysis System (SAS, SAS Institute Inc., Cary, NC, USA). The cleavage rate of pST101 cell and control cell derived SCNT embryos were 83.6 ± 2.0% (267/318) and 84.0 ± 2.2% (139/167) respectively, there were no significant (P > 0.05) differences between them. The blastocyst rate from pST101 cell and control cell derived SCNT embryos were 21.4 ± 1.8% (65/318) and 15.0 ± 3.0% (26/167) respectively, the SCNT embryos derived from pST101 cell showed a higher (P < 0.05) blastocyst rate. These results indicated that follistatin transgenic porcine embryo can be successfully constructed and can develop in vitro to blastocyst with a higher rate than control SCNT embryo. Further study will be focused on the full development in vivo of the transgenic embryos and phenotype of transgenic piglets.

This work was supported in part by the National High-tech Research and Development Program of China (2008AA101006) and Major Projects for Transgenic Animals of China (2008ZX08006-002).
94 EFFECT OF OVARIAN SIZE ON THE VIABILITY OF CRYOPRESERVED SYRIAN HAMSTER OVARIES
Authors: Suzuki O. , Koura M. , Noguchi Y. , Uchio-Yamada K. , Matsuda J.
Published: August 2011

Ovaries can be used in place of gametes as a resource for cryopreservation. Thus far, we have shown that in the Syrian hamster, pups can be successfully produced through the transplantation of fresh and cryopreserved ovaries. In this study, we attempted to simplify the ovarian cryopreservation procedures to facilitate the easy application of these techniques. In particular, the procedure would be easier and quicker if the ovaries did not have to be cut into small pieces before ovarian cryopreservation. Therefore, we examined the effects of ovarian fragment size on subsequent offspring production. Ovaries were cryopreserved by vitrification according to methods described for mouse ovaries (Migishima et al. 2003 Biol. Reprod. 68, 881–887). In brief, ovaries were collected from 3-week-old J2N-n strain (white coat) females. Because we had previously confirmed that quarter-sized ovaries could be used for cryopreservation, intact (∼3.5 × 2 mm) and half-sliced ovaries (∼2 × 1.5 mm) were cryopreserved by vitrification using DAP213 solution (2 M dimethyl sulfoxide, 1 M acetamide, and 3 M propylene glycol; Nakagata 1989 J. Reprod. Fertil. 87, 479–483). Thawed ovaries were transferred into the ovarian bursae (single intact ovary or two half-sliced ovarian pieces per ovarian bursa) of 3-week-old recipient females with agouti-coat (Slc:Syrian) under isoflurane anaesthesia. Not all of the recipients’ ovaries were removed immediately before transfer; both the donor’s and a small portion of the recipient’s ovaries were present in each ovarian bursa. Successful ovarian transplantation was verified by examining the coat colour (non-agouti colour) of offspring produced in test matings between mature recipients and J2N-n males. Of the 3 and 6 recipients that received intact and sliced ovaries, respectively, 2 and 5 recipients delivered pups. Based on the coat colour of the pups, 1 of the 5 recipients delivered a pup produced from transplanted half-sliced ovaries. Our results indicate that half-sliced-size ovaries (smaller than a 2-mm cube) can be successfully used for cryopreservation in the Syrian hamster. Specifically, it is not required to slice the ovary into smaller pieces.

This work was supported by a grant from the Ministry of Health, Labour and Welfare, Japan and Grant-in-Aid for Scientific Research (C).
Conservation risks of exotic chukars (Alectoris chukar) and their associated management: implications for a widely introduced phasianid
Authors: Larsen Randy T. , Flinders Jerran T. , Mitchell Dean L. , Perkins Ernest R.
Published: August 2011

Chukars (Alectoris chukar) have been widely introduced throughout the world. Their introductions and associated management for sport hunting have the potential to affect native ecosystems in a variety of ways. Our specific objectives were: (1) to document species using water developments designed to benefit chukar populations to determine whether, and at what prevalence, exotic species appear to use, and presumably benefit from, additional watering points; (2) to describe chukar diet with specific reference to cheatgrass and other exotic plant seeds; and (3) to determine whether chukars are a likely vector for dispersal of cheatgrass and/or other plant seeds via passage through the gut. In total, 27 different wildlife species were photographed across all 36 sampled water developments. Three exotic species were photographed to include chukars, rock dove (Columba livia), and red fox (Vulpes vulpes), with the latter two species photographed at only two and one site respectively. Mean number of species photographed (5.69 ± 1.09) ranged from 1 to 13, but was estimated near 10 after accounting for sampling time. Cheatgrass seed was found in 76.3% of crops and constituted 45.2% of dry weight. Thirteen plants germinated from 503 chukar faecal droppings. We found no evidence of widespread use of water points designed for chukars by other exotic species or dispersal of cheatgrass seed via passage through the gut. Chukars appear (at least initially) benign and they are not likely to be major vectors in plant seed dispersal. Furthermore, chukars could foster localised plant diversity in that they consume large quantities of primarily exotic plant seed and do not show a propensity for dispersal of seeds through faecal droppings.
Gluconacetobacter kakiaceti sp. nov., an acetic acid bacterium isolated from a traditional Japanese fruit vinegar
Authors: Iino Takao , Suzuki Rei , Tanaka Naoto , Kosako Yoshimasa , Ohkuma Moriya , Komagata Kazuo , Uchimura Tai
Published: August 2011

Two novel acetic acid bacteria, strains G5-1T and I5-1, were isolated from traditional kaki vinegar (produced from fruits of kaki, Diospyros kaki Thunb.), collected in Kumamoto Prefecture, Japan. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains G5-1T and I5-1 formed a distinct subline in the genus

Gluconacetobacter

and were closely related to

Gluconacetobacter swingsii

DST GL01T (99.3 % 16S rRNA gene sequence similarity). The isolates showed 96–100 % DNA–DNA relatedness with each other, but <53 % DNA–DNA relatedness with closely related members of the genus

Gluconacetobacter

. The isolates could be distinguished from closely related members of the genus

Gluconacetobacter

by not producing 2- and 5-ketogluconic acids from glucose, producing cellulose, growing without acetic acid and with 30 % (w/v) d-glucose, and producing acid from sugars and alcohols. Furthermore, the genomic DNA G+C contents of strains G5-1T and I5-1 were a little higher than those of their closest phylogenetic neighbours. On the basis of the phenotypic characteristics and phylogenetic position, strains G5-1T and I5-1 are assigned to a novel species, for which the name Gluconacetobacter kakiaceti sp. nov. is proposed; the type strain is G5-1T ( = JCM 25156T = NRIC 0798T = LMG 26206T).
Winogradskyella ulvae sp. nov., an epiphyte of a Pacific seaweed, and emended descriptions of the genus

Winogradskyella

and

Winogradskyella thalassocola

,

Winogradskyella echinorum

,

Winogradskyella exilis

and

Winogradskyella eximia
Authors: Nedashkovskaya Olga I. , Kukhlevskiy Andrey D. , Zhukova Natalia V.
Published: August 2011

A Gram-negative, facultatively anaerobic, slightly halophilic bacterium that was motile by gliding, designated KMM 6390T, was isolated from the coastal green alga Ulva fenestrata collected from the Sea of Japan. A comparative analysis of 16S rRNA gene sequences revealed that strain KMM 6390T was a member of the genus

Winogradskyella

, with

Winogradskyella echinorum

KMM 6211T as its closest relative (97.8 % 16S rRNA gene sequence similarity). 16S rRNA gene sequence similarities between the isolate and other members of the genus

Winogradskyella

were 95.4–96.7 %. DNA–DNA relatedness between the isolate and

W. echinorum

KMM 6211T was 27 %. Strain KMM 6390T was able to grow with 0.5–4.0 % NaCl and at 4–37 °C. The isolate decomposed casein, gelatin, starch and DNA and produced acid from galactose, d-glucose, lactose and maltose. The major fatty acids were iso-C15 : 0, iso-C15 : 1, anteiso-C15 : 0 and C15 : 0. The polar lipid profile comprised phosphatidylethanolamine and two unknown aminolipids. The DNA G+C content was 34.2 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic investigations, strain KMM 6390T represents a novel species of the genus

Winogradskyella

, for which the name Winogradskyella ulvae sp. nov. is proposed; the type strain is KMM 6390T ( = KCTC 23626T = LMG 26444T). Emended descriptions of the genus

Winogradskyella

and of

W. thalassocola

,

W. echinorum

,

W. exilis

and

W. eximia

are also provided.
Williamsia limnetica sp. nov., isolated from a limnetic lake sediment
Authors: Sazak Anil , Sahin Nevzat
Published: August 2011

An actinomycete, strain L1505T, was isolated from a limnetic lake sediment and found to have morphological, biochemical, physiological and chemotaxonomic properties consistent with its classification in the genus

Williamsia

. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain L1505T formed a distinct lineage within the genus

Williamsia

. The isolate belonged to a cluster containing W. muralis
MA140/96T, W. marianensis
MT8T and W. faeni
N1350T, with which the isolate shared 99.0–98.2 % 16S rRNA gene sequence similarity. Genotypic and phenotypic data also indicated that the isolate was different from known members of the genus

Williamsia

. On the basis of these data, strain 1505T is considered to represent a novel species of the genus

Williamsia

, for which the name Williamsia limnetica sp. nov. is proposed (type strain L1505T = DSM 45521T = NRRL B-24829T = KCTC 19981T).
Kloeckera taiwanica sp. nov., an ascomycetous apiculate yeast species isolated from mushroom fruiting bodies
Authors: Chang Chin-Feng , Huang Li-Yin , Chen Shan-Fu , Lee Ching-Fu
Published: August 2011

Three apiculate yeast strains, EJ7M09T, GJ5M15 and GJ15M04, isolated from mushrooms in Taiwan were found to represent a novel species of the genus Kloeckera. The phylogenetically closest relative of this novel species is Hanseniaspora occidentalis, but the type strain of H. occidentalis differed by 4.6 % divergence (25 substitutions; 5 gaps) in the sequence of the D1/D2 domain of the large subunit rRNA gene. This difference clearly suggests that the three strains represent a distinct species. As none of the strains that were examined in this study produced ascospores or exhibited conjugation on common sporulation medium either alone or in a pairwise mixture, this species could be considered as an anamorphic member of the genus Hanseniaspora, and a novel species, Kloeckera taiwanica sp. nov., is proposed, with EJ7M09T ( = BCRC 23182T = CBS 11434T) as the type strain.
Classification of

Pasteurella

species B as Pasteurella oralis sp. nov.
Authors: Christensen Henrik , Bertelsen Mads F. , Bojesen Anders Miki , Bisgaard Magne
Published: August 2011

Pasteurella

species B has so far only been reported from the oral cavity of dogs, cats and a ferret. In the present study, information from 15 recent isolates from different sources, including African hedgehogs (Atelerix albiventris), banded mongoose (Mungos mungo), Moholi bushbabies (Galago moholi) and pneumonia of a cat, were compared to five strains investigated previously from bite wounds in humans inflicted by a cat and dog and from gingiva of a cat. rpoB gene sequence comparison showed that 17 isolates, including the reference strain (CCUG 19794T), had identical sequences, whereas two were closely related and demonstrated 97.9 and 99.6 % similarity to strain CCUG 19794T, respectively; the type strain of

Pasteurella stomatis

was the most closely related strain, with 92.3 % similarity. This is within the mean range (76–100 %) of rpoB gene sequence similarity between species of the same genus within the family

Pasteurellaceae

. 16S rRNA gene sequencing of four strains selected based on rpoB sequence comparison showed at least 99.7 % similarity between strains of

Pasteurella

species B, with 96.2 % similarity to the type strain of the closest related species (

Pasteurella canis

), indicating that

Pasteurella

species B should have separate species status. Separate species status was also documented when recN sequence comparisons were converted to a genome similarity of 93.7 % within

Pasteurella

species B and 59.0 % to the type strain of the closest related species (

P. canis

). Based on analysis of the phylogenetic and phenotypic data, and since most isolates originate from the oral cavities of a diverse group of animals, it is suggested that these bacteria be classified as Pasteurella oralis sp. nov.; the type strain is P683T ( = CCUG 19794T = CCM 7950T = strain 23193T = MCCM 00102T), obtained from a cat. Previous reports of the type strain have shown ubiquinone-8, demethylmenaquinone-8 and menaquinone-8 as the major quinones. Polyamines in the type strain were reported as diaminopropane, putrescine, cadaverine, sym-norspermidine, spermidine and spermine in a previous investigation, and the major fatty acids of the type strain were reported to be C16 : 0, C16 : 1ω7c and C14 : 0, with minor amounts of C18 : 0 and C18 : 1ω9c. The DNA G+C content of the type strain has been reported to be 40.0 mol%.
An Emerging Role for the Delta Opioid Receptor in the Regulation of Mu Opioid Receptor Function
Authors: Rozenfeld Raphael , Abul-Husn Noura S. , Gomez Ivone , Devi Lakshmi A.
Published: August 2011

Morphine and related opiates are commonly used in the clinical management of various types of pain. However, the antinociceptive properties of morphine are often overshadowed by the development of tolerance and dependence following its chronic use. The mechanisms underlying opiate tolerance are not fully understood, but appear to involve numerous and complex physiological adaptations. Recently, a role for the heterodimerization of mu and delta opioid receptors in the development of morphine tolerance has been proposed. This novel mechanism could help us to understand several observations, such as the critical role of delta opioid receptor regulation, the impact of delta opioid receptor binding site occupancy, and the participation of beta-arrestin2, in the development of morphine tolerance.