bluuebunny/crossref_metadata_2025 huggingface dataset:
Significant population genetic structure of yellowfin seabream Acanthopagrus latus in China
Authors: Xia J.‐H. , Huang J.‐H. , Gong J.‐B. , Jiang S.‐G.
Published: November 2008

The yellowfin seabream Acanthopagrus latus is widely distributed throughout the Indo‐West Pacific. The genetic analyses of mtDNA control region sequence variation in samples from Chinese waters revealed a pattern of genetic structure between southern and northern samples (P < 0·05) and high levels of genetic diversity in this species. Significant isolation by distance between southern and northern locations (Mantel test r = 0·35, P = 0·04) and among northern populations (r = 0·55, P = 0·01) indicated that ocean straits and other barriers substantially limit gene flow. Phylogeographical analysis also revealed two major divergent mtDNA lineages across Chinese waters. For conservation, A. latus populations in Chinese waters should be divided into at least two management units for protection.
The evolution of teleost pigmentation and the fish‐specific genome duplication
Authors: Braasch I. , Volff J.‐N. , Schartl M.
Published: November 2008

Teleost fishes have evolved a unique complexity and diversity of pigmentation and colour patterning that is unmatched among vertebrates. Teleost colouration is mediated by five different major types of neural‐crest derived pigment cells, while tetrapods have a smaller repertoire of such chromatophores. The genetic basis of teleost colouration has been mainly uncovered by the cloning of pigmentation genes in mutants of zebrafishDanio rerioand medakaOryzias latipes. Many of these teleost pigmentation genes were already known as key players in mammalian pigmentation, suggesting partial conservation of the corresponding developmental programme among vertebrates. Strikingly, teleost fishes have additional copies of many pigmentation genes compared with tetrapods, mainly as a result of a whole‐genome duplication that occurred 320–350 million years ago at the base of the teleost lineage, the so‐called fish‐specific genome duplication. Furthermore, teleosts have retained several duplicated pigmentation genes from earlier rounds of genome duplication in the vertebrate lineage, which were lost in other vertebrate groups. It was hypothesized that divergent evolution of such duplicated genes may have played an important role in pigmentation diversity and complexity in teleost fishes, which therefore not only provide important insights into the evolution of the vertebrate pigmentary system but also allow us to study the significance of genome duplications for vertebrate biodiversity.
A new species of the genus Mekongina Fowler, 1937 (Cypriniformes: Cyprinidae) from South China
Authors: Yang J. , Chen X. Y. , Yang J. X.
Published: November 2008

A new species of cyprinid fish, Mekongina lancangensis, is described from the upper Mekong River drainage in Southern Yunnan, China. The new species is distinguished from the other species of Mekongina occurring in the lower Mekong River drainage by possessing the following combination of characters: one pair of rostral barbels; two rows of tubercles irregularly scattered on the snout and cheeks, with two enlarged tubercles present at each side of anterior of the snout; 19–27 rostral marginal lappets; lateral line with 38–41 scales; 5·5 or 6·5 scales in transverse series from dorsal‐fin origin to lateral line; 18–20 circumpeduncular scales; snout length 31·9–36·9% head length; tip of depressed anal‐fin rays extending to the caudal‐fin base.
Genetic divergence between sympatric Arctic charr Salvelinus alpinus morphs in Gander Lake, Newfoundland: roles of migration, mutation and unequal effective population sizes
Authors: Gomez‐Uchida D. , Dunphy K. P. , O’Connell M. F. , Ruzzante D. E.
Published: November 2008

A suite of 10 microsatellite loci was used to examine genetic divergence between two sympatric morphs of Arctic charr Salvelinus alpinus (‘dark’ and ‘pale’) inhabiting Gander Lake, Newfoundland. Results can be summarized as follows: (1) the morphs are strongly reproductively isolated – gene flow–migration estimates were consistently low in long and short‐term evolutionary timescales of analysis; (2) intermorph divergence based on allele size (RST) was significantly larger than those based on allele state (θ) implying a cumulative effect of stepwise‐like mutations; (3) historical (coalescent) and current (linkage disequilibrium) point estimates of effective population size (Ne) were consistently higher for dark than for pale S. alpinus. The first and second findings lend support to the hypothesis that divergence between forms may have preceded the last glacial period (ending c. 12 000 years bp). The third finding argues for significant differences in habitat quantity and quality between morphs, which were emphasized in a previous study. Overall, these analyses underscore the importance of genetic assessment and monitoring in the conservation of fish diversity, with emphasis on ‘rare’ or under‐represented forms among temperate species pairs.
Evidence of multiple paternity in spiny dogfish (Squalus acanthias) broods based on microsatellite analysis
Authors: Lage C. R. , Petersen C. W. , Forest D. , Barnes D. , Kornfield I. , Wray C.
Published: November 2008

An understanding of alternative mating systems is a fundamental requirement for the effective management of vulnerable species. Microsatellite analysis of spiny dogfish Squalus acanthias broods from the north‐west Atlantic provides novel evidence of polyandry in this species.
Lack of genetic differentiation in the fat snook Centropomus parallelus (Teleostei: Centropomidae) along the Brazilian coast
Authors: Prodocimo V. , TschÁ M. K. , Pie M. R. , Oliveira‐Neto J. F. , Ostrensky A. , Boeger W. A.
Published: November 2008

The patterns of genetic variation in a fragment of the cytochrome b mtDNA gene in 130 individuals of the fat snook (Centropomus parallelus) did not show evidence of genetic differentiation along the Brazilian coast as shown in F‐statistics and AMOVA analyses, indicating the existence of high gene flow among the studied populations. In addition, the study of the evolutionary demography of the species using mismatch distribution analysis and statistical tests such as Tajima’s D and Fu’s F indicate that it experienced a pattern of population expansion during its recent evolutionary history.
Engagement of Penicillium marneffei conidia with multiple pattern recognition receptors on human monocytes
Authors: Srinoulprasert Yuttana , Pongtanalert Piyapong , Chawengkirttikul Runglawan , Chaiyaroj Sansanee C.
Published: November 2008

P. marneffei is a thermal dimorphic fungus which causes penicilliosis, an opportunistic infection in immunocompromised patients in South and Southeast Asia. Little is known about the innate immune response to P. marneffei infection. Therefore, the initial response of macrophages to P. marneffei conidia was evaluated by us. Adhesion between monocytes from healthy humans and fungal conidia was examined and found to be specifically inhibited by MAbs against PRR, such as MR, (TLR)1, TLR2, TLR4, TLR6, CD14, CD11a, CD11b, and CD18. To study the consequences of these interactions, cytokines were also examined by ELISA. Binding of P. marneffei conidia to monocytes was significantly inhibited, in a dose‐dependent manner, by MAbs against MR, TLR1, TLR2, TLR4, TLR6, CD14, CD11b and CD18. When monocytes were co‐cultured with the conidia, there was an increase in the amount of surface CD40 and CD86 expression, together with TNF‐α and IL‐1β production, compared to unstimulated controls. In assays containing anti‐TLR4 or anti‐CD14 antibody, reduction in the amount of TNF‐α released by monocytes stimulated with P. marneffei conidia was detected. In addition, it was found that production of TNF‐α and IL‐1β from adherent peripheral blood monocytes was partially impaired when heat‐inactivated autologous serum, in place of untreated autologous serum, was added to the assay. These results demonstrate that various PRR on human monocytes participate in the initial recognition of P. marneffei conidia, and the engagement of PRR could partly initiate proinflammatory cytokine production.
The influence of light and temperature on the germination of two Ugandan medicinal trees
Authors: Stangeland Torunn , Tabuti John R. S. , Lye Kåre A.
Published: November 2008

For reasons of the problems of establishment of some Ugandan trees in certain environments, we investigated the influence of temperature and light on germination and seedling growth of two locally threatened medicinal trees, Hallea rubrostipulata and Sarcocephalus latifolius, to facilitate their establishment. Field and controlled laboratory experiments were carried out to investigate the species germination requirements and seedling growth. Both species needed light to germinate. Hallea rubrostipulata had a temperature optimum of 25°C with 79% germination, while for S. latifolius, the total germination after 28 days was close to 60% at temperatures from 20 to 35°C. Seedlings of S. latifolius survived well at 35°C, while those of H. rubrostipulata died at this high temperature. Conversely, seedling of S. latifolius died at the low temperature of 15°C. However, in field experiment S. latifolius failed to germinate in the available degraded environments, probably because of predation and because the soil is not able to retain water long enough to support seedling growth. We, therefore, conclude that in this part of Uganda, nursery assistance is needed to establish healthy populations of Sarcocephalus and many other endangered trees.
Amplified in breast cancer 1 expression in breast cancer
Authors: Thorat M A , Turbin D , Morimiya A , Leung S , Zhang Q , Jeng M‐H , Huntsman D G , Nakshatri H , Badve S
Published: November 2008

Aims:  The amplified in breast cancer 1 (AIB1), steroid receptor co‐activator family member, acts as an oestrogen receptor (ER) co‐activator. Acting with HER‐2, it is thought to play a role in endocrine resistance by facilitating ER–growth factor crosstalk. The aim was to analyse AIB1 expression by immunohistochemistry and study its correlations with other prognostic variables in breast cancer and its effect on survival.

Methods:  A tissue microarray comprising tumours from 438 patients with 15.4 years’ median follow‐up was used. Interpretable AIB1 expression obtained in 395 patients was analysed along with other prognostic factors in breast cancer.

Results:  AIB1 expression scores ranged from 0 to 30; positive AIB1 expression (score > 14) was seen in 146/395 breast cancers; it correlated negatively with ER (P = 0.003) and progesterone receptor (PR) (P = 0.007), and positively with HER‐2 (P = 0.005) and tumour grade (P = 0.014). It did not correlate with nodal status (P = 0.437). Among ER+ patients, AIB1 expression showed a trend towards loss of PR expression (29% versus 20%; P = 0.14). AIB1 did not predict survival on univariate or multivariate analysis.

Conclusions:  AIB1 expression correlates with HER‐2 expression in breast cancer and shows a trend of association with loss of PR expression in ER+ tumours. Our study supports the postulated role of AIB1 in ER–growth factor interactions.
Quality aspects of the tissue microarray technique in a population‐based cohort with ductal carcinoma in situ of the breast
Authors: Wärnberg F , Amini R‐M , Goldman M , Jirström K
Published: November 2008

Aims:  Tissue microarray (TMA) is an efficient technique for analysis of molecular markers. Prospectively collected samples have been reported to give excellent concordance between TMA data and corresponding whole‐sections. The aim was to evaluate the usefulness of TMA in a population‐based cohort of 213 women with ductal carcinoma in situ of the breast (DCIS).

Methods and results:  We studied immunohistochemical HER2, oestrogen (ER) and progesterone (PR) receptor status. The prognostic impact was similar for all markers comparing whole sections and TMAs. The proportion of positive tumours was similar regarding HER2 and ER, whereas PR tumours were more frequently positive in the TMAs (P = 0.007). The concordance was 80% (κ value 0.63) between original sections and TMAs. The proportion of successfully analysed tumours was 70%. Smaller tumours had a lower ratio (P < 0.0001) and a larger proportion of mismatched results (P = 0.05).

Conclusions:  Retrospective analyses of tumours from cohorts with long‐term follow‐up are indispensable. We have shown that the TMA technique is a useful tool for high‐throughput analysis of DCIS. However, our study has pinpointed some technical hazards within a population‐based cohort, including many small lesions and the poor condition of some donor blocks. Mismatched results may be due to tumour heterogeneity.
Predictive value of needle core biopsy diagnoses of lesions of uncertain malignant potential (B3) in abnormalities detected by mammographic screening
Authors: El‐Sayed M E , Rakha E A , Reed J , Lee A H S , Evans A J , Ellis I O
Published: November 2008

Aims:  Breast needle core biopsy (NCB) is now a commonplace diagnostic procedure in breast cancer screening, providing accurate diagnoses of both benign and malignant lesions. However, NCB may result in the borderline diagnoses of lesion of uncertain malignant potential (B3) or suspicious of malignancy (B4). The aim was to study a large series of B3 cases from population‐based screening subjects in order to evaluate positive predictive values (PPVs) for malignancy.

Methods and results:  The results of 523 NCBs of women screened over a 7‐year period (1999–2006) in the East Midlands region, UK, with a B3 diagnosis who underwent surgical excision, were reviewed and compared with the final excision histology. Five percent of NCBs were reported as B3. The most frequent histological subtypes were atypical intraductal epithelial proliferation (AIDEP) and radial scar/complex sclerosing lesion (RS/CSL). Final excision histology was benign in 417 (80%) and malignant in 106 (20%) subjects (60 ductal carcinoma in situ and 46 invasive carcinoma). Lesion‐specific PPVs were as follows: AIDEP 32%; lobular neoplasia (LN) 30%; RS/CSL with AIDEP or LN 24%; RS/CSL without atypia 9%; papillary lesion with AIDEP or LN 36%; and papillary lesion without atypia 4%. Five of the 32 fibroepithelial lesions with cellular stroma were phyllodes tumours (four benign and one borderline). None of the five mucinous lesions on NCB was malignant.

Conclusions:  Our results show that approximately one‐fifth of NCB of screen‐detected breast lesions classified as B3 are malignant on excision, and the likelihood of malignancy varies substantially between different histological subtypes.
Intratumoural heterogeneity of intestinal expression reflects environmental induction and progression‐related loss of induction in undifferentiated‐type gastric carcinomas
Authors: Natsagdorj L , Sugihara H , Bamba M , Hattori T
Published: November 2008

Aims:  Gene expression in tumours is regulated by environmental as well as genetic/epigenetic factors. This study assessed the environmental factors in intestinal expression of gastric cancers.

Methods and results:  We immunohistochemically examined intratumoural heterogeneity in the expression of Cdx2, MUC2, MUC5AC and MUC6 in 39 intramucosal and 49 extramucosally invasive undifferentiated‐type gastric carcinomas (UGCs), consisting of signet ring cell carcinomas showing a layered structure (LS) in the mucosa and dedifferentiated tubular adenocarcinomas without LS and with minor tubular components (TC). The LS retains mucosal vertical polarity with superficial MUC5AC expression. Loss of this polarity was independent of intestinal expression and associated with extramucosal invasion. In LS+ UGCs, intestinal expression was enhanced as the size of mucosal spread increased and was significantly reduced with deeper extramucosal invasion, whereas, in LS−/TC+ UGCs, intestinal expression was frequent and predominant in the mucosa and was insignificantly reduced with deeper extramucosal invasion.

Conclusions:  In LS+ UGCs, intestinal expression showed dynamic alteration probably by environmental induction and progression‐related loss of induction, whereas it was relatively stable in LS−/TC+ UGCs. Thus, intestinal expression in UGCs is not useful as a marker of tumour progression because it is also affected by environmental factors and genetic lineage.
Annexin A1 subcellular expression in laryngeal squamous cell carcinoma
Authors: Alves V A F , Nonogaki S , Cury P M , Wünsch‐Filho V , De Carvalho M B , Michaluart‐Júnior P , Moyses R A , Curioni O A , Figueiredo D L A , Scapulatempo‐Neto C , Parra E R , Polachini G M , Silistino‐Souza R , Oliani S M , Silva‐Júnior W A , Nobrega F G ,   Head and Neck Genome Project/GENCAPO, Tajara E H , Zago M A
Published: November 2008

Aims:  Annexin A1 (ANXA1) is a soluble cytoplasmic protein, moving to membranes when calcium levels are elevated. ANXA1 has also been shown to move to the nucleus or outside the cells, depending on tyrosine‐kinase signalling, thus interfering in cytoskeletal organization and cell differentiation, mostly in inflammatory and neoplastic processes. The aim was to investigate subcellular patterns of immunohistochemical expression of ANXA1 in neoplastic and non‐neoplastic samples from patients with laryngeal squamous cell carcinomas (LSCC), to elucidate the role of ANXA1 in laryngeal carcinogenesis.

Methods and results:  Serial analysis of gene expression experiments detected reduced expression of ANXA1 gene in LSCC compared with the corresponding non‐neoplastic margins. Quantitative polymerase chain reaction confirmed ANXA1 low expression in 15 LSCC and eight matched normal samples. Thus, we investigated subcellular patterns of immunohistochemical expression of ANXA1 in 241 paraffin‐embedded samples from 95 patients with LSCC. The results showed ANXA1 down‐regulation in dysplastic, tumourous and metastatic lesions and provided evidence for the progressive migration of ANXA1 from the nucleus towards the membrane during laryngeal tumorigenesis.

Conclusions:  ANXA1 dysregulation was observed early in laryngeal carcinogenesis, in intra‐epithelial neoplasms; it was not found related to prognostic parameters, such as nodal metastases.
Comparison of histological parameters for the diagnosis of eosinophilic oesophagitis versus gastro‐oesophageal reflux disease on oesophageal biopsy material
Authors: Mueller S , Neureiter D , Aigner T , Stolte M
Published: November 2008

Aims:  Eosinophil infiltration of the oesophageal epithelium is the cardinal pathomorphological finding in eosinophilic oesophagitis (EO), but gastro‐oesophageal reflux disease (GORD) is also associated with increased eosinophils. The aim was to compare histological parameters for the diagnosis of EO versus GORD on routinely taken biopsy specimens.

Methods and results:  One hundred and five routine biopsy specimens with EO (n = 62), GORD (n = 24) and probable EO (n = 19) from 74 patients (52 men, 22 women; mean age 43.7 years) were analysed for numbers of eosinophils, mast cells, degranulation and qualitative changes of oesophageal epithelium using immunohistochemistry with monoclonal antibodies against eosinophil peroxidase and eosinophil major basic protein and mast cell tryptase. Eosinophil infiltration was significantly higher in EO than in GORD both on haematoxylin and eosin staining (54.8 versus 9.1; P < 0.05) and immunohistochemistry (77.5 versus 24.7; P < 0.05). Eosinophil degranulation was significantly more intense in EO than in GORD (1.16 versus 0.41; P < 0.05). Furthermore, eosinophilia‐codependent secondary qualitative changes of squamous epithelium in EO were generally more extensive than those in GORD.

Conclusions:  Histological differential diagnosis of EO and GORD should be based on eosinophil counts, secondary morphological changes of eosinophils and oesophageal squamous epithelium, especially in cases suspicious of EO.
Reg IV expression is associated with cell growth and prognosis of adenoid cystic carcinoma in the salivary gland
Authors: Sasahira T , Oue N , Kirita T , Luo Y , Bhawal U K , Fujii K , Yasui W , Kuniyasu H
Published: November 2008

Aims:  Regenerating islet‐derived family, member 4 (Reg IV) is associated with the progression of various cancers. The aim was to examine Reg IV expression in adenoid cystic carcinomas (ACCs) in salivary glands.

Methods and results:  Reg IV expression was detected by immunohistochemistry and compared with clinicopathological parameters. Expression of phosphorylated epidermal growth factor receptor (pEGFR), phosphorylated AKT (pAKT) and MUC2 was examined by immunohistochemistry. Reg IV function was assessed with Reg IV antisense S‐oligodeoxynucleotides (AS) in ACC3 human ACC cells. Reg IV was expressed by salivary duct epithelia and acinus myoepithelia, but not in squamous epithelia. Reg IV expression was found in 41% (17/41) of ACCs, but in none of 40 oral squamous cell carcinomas (OSCCs) and was associated with nodal metastasis (P = 0.047) and poor prognosis (P = 0.012) in ACCs. Reg IV expression was associated with pEGFR (14/17, 82%) in Reg IV+ ACCs, but had no relationship with pAKT or MUC2 expression in ACCs. Cell growth was inhibited by AS treatment in Reg IV+ ACC3 cells, but not in HSC‐4 OSCC cells, whereas in vitro invasion of neither cell types was affected by AS treatment.

Conclusions:  These results suggest that Reg IV might accelerate cell growth and disease progression of ACCs.
Effect of gamma irradiation and sulphitation treatments on keeping quality of white button mushroom Agaricus bisporus (J. Lge)
Authors: Wani Ali M. , Hussain Peerzada R. , Meena Raghuveer S. , Dar Mohmad A. , Mir Mohmad A.
Published: November 2008

Gamma irradiation, alone and in combination with sulphitation, was tested for preventing the browning and maintaining the quality attributes of the white button mushrooms. Mushrooms were subjected to treatment of gamma irradiation in the dose range of 0.5–2.0 kGy and to combination treatments of sulphitation at a concentration of 0.1% potassium metabisulphite (KMS) and gamma irradiation (dose range 0.5–2.0 kGy) followed by storage at 10 ± 2 °C (RH 85%). A dose of 2.0 kGy significantly reduced the weight loss, prevented browning and mould growth. Cap and veil opening of mushrooms was delayed by 9 days and shelf life was extended by 12 days at a dose level of 2.0 kGy. Sulphitation alone at a concentration of 0.1% KMS was effective in controlling browning only upto 3 days, beyond which both browning and cap opening increased significantly (P ≤ 0.05) and the samples were unacceptable after 6 days of storage. No synergistic effect of sulphitation and irradiation was observed with respect to the shelf‐life extension of mushroom.
Novel microwave–freeze drying of onion slices
Authors: Abbasi Soleiman , Azari Sara
Published: November 2008

The present study aimed to introduce the capability of a novel dehydration technique. To do so, slices with various thicknesses (3.5 and 7 mm) of white onions were dried using commercial freeze drier (abs. pressure 0.005 mbar, temp. 45 °C), our own designed microwave‐vacuum drier (abs. pressure down to 300 mbar) under various microwave powers (120–1200 W) and microwave‐vacuum–freeze drier (onion slices kept at −20 °C for 2 h). Then, their dehydration rates and some quality parameters, such as rehydration ratio, colour (L*, a* and b*) and micro‐structure were investigated. Our findings showed that microwave‐vacuum–freeze drier is practically a rapid, simple, efficient, economic and novel dehydration technique which can be used for dehydration of mainly foodstuffs. The quality properties of slices produced by this novel method were also completely comparable and competitive with commercial freeze drier with over 96% saving on processing time and enormous amount on energy and capital investments.
Significance of summer fog and overcast for drought stress and ecological functioning of coastal California endemic plant species
Authors: Fischer Douglas T. , Still Christopher J. , Williams A. Park
Published: November 2008

Aim Fog drip is a crucial water source for plants in many ecosystems, including a number of global biodiversity hotspots. In California, dozens of rare, drought‐sensitive plant species are endemic to coastal areas where the dominant summer moisture source is fog. Low clouds that provide water to these semi‐arid ecosystems through fog drip can also sharply reduce evaporative water losses by providing shade. We quantified the relative hydrological importance of cloud shading vs. fog drip. We then examined how both factors influence the range dynamics of an apparently fog‐dependent plant species spanning a small‐scale cloud gradient.

Location The study area is on Santa Cruz Island off the coast of southern California. It is near the southern range limit of bishop pine (Pinus muricataD. Don), a tree endemic to the coasts of California and Baja, Mexico.

Methods We measured climate across a pine stand along a 7 km, coastal–inland elevation transect. Short‐term (1–5 years) monitoring and remote sensing data revealed strong climatic gradients driven primarily by cloud cover. Long‐term (102 years) effects of these gradients were estimated using a water balance model.

Results We found that shade from persistent low clouds near the coast reduced annual drought stress by 22–40% compared with clearer conditions further inland. Fog drip at higher elevations provided sufficient extra water to reduce annual drought stress by 20–36%. Sites located at both high elevation and nearer the coast were subject to both effects. Together, these effects reduced average annual drought stress by 56% and dramatically reduced the frequency of severe drought over the last century. At lower elevation (without appreciable fog drip) and also near the inland edge of the stand (with less cloud shading) severe droughts episodically kill most pine recruits, thereby limiting the local range of this species.

Main conclusions Persistent cloud shading can influence hydrology as much as fog drip in cloud‐affected ecosystems. Understanding the patterns of both cloud shading and fog drip and their respective impacts on ecosystem water budgets is necessary to fully understand past species range shifts and to anticipate future climate change‐induced range shifts in fog‐dependent ecosystems.
A hawkmoth crossroads? Species richness, seasonality and biogeographical affinities of Sphingidae in a Brazilian Cerrado
Authors: Amorim Felipe W. , DeÁvila Jr Rubem S. , De Camargo Amabílio J. A. , Vieira Antenor L. , Oliveira Paulo E.
Published: November 2008

Aim  The aim of the study was (1) to describe the biodiversity of the sphingid assemblage in a Cerrado area in the Triângulo Mineiro region, south‐east of Brazil; (2) to evaluate the seasonal variations in species composition; (3) to compare the faunistic relationships between the Cerrado biome and adjacent ecosystems; and (4) to analyse the biogeographical pattern of species distribution in the Neotropical region in a historical context.

Location  Panga Ecological Station (PES), 30 km south of the city of Uberlândia, and other areas of the Triângulo Mineiro region, Minas Gerais state, south‐eastern Brazil.

Methods  Moth richness and abundance were monitored monthly at the PES from August 2003 to July 2004, with additional collections at this locality in 2001/2002, 2005 and 2006. Complementary moth richness and abundance data were also collected in other areas of the Triângulo Mineiro region. All collections were made using light traps, and the hawkmoths were mounted and identified. Cluster analysis, rarefaction curves and estimators of total species richness were used to compare the Cerrado hawkmoth assemblage with assemblages derived from other surveys in the Neotropics.

Results  In total, 61 hawkmoth species were recorded for the study region and their occurrence was markedly seasonal. The hawkmoth assemblage in the study area presented the closest similarity with rain forest areas and with a tropical dry forest area in Central America. The area shared species with both rain forest and seasonally dry tropical forest (SDTF) ecosystems, including supposedly endemic species previously recorded only in the latter areas. Rarefaction curves and estimators of the total number of species showed species richness to be comparable with other highly diverse forest areas in the Neotropics, such as the Brazilian Amazon and Costa Rica.

Main conclusions  This short‐term study is the first systematic survey of hawkmoths in the Cerrado. It has recorded around 22% of the South American fauna and highlighted the high species richness of the region, which compares favourably with that in other rain forest ecosystems. The survey indicates high regional diversity, and has shown that the Cerrado harbours a hawkmoth fauna comprising both rain forest elements, probably distributed along humid gallery forest corridors, and SDTF elements, supporting the idea of a historical Pleistocene arc connecting the Caatinga domain and other seasonal dry forest areas across the Cerrado region.
A comparative neuropsychological test battery differentiates cognitive signatures of Fragile X and Down syndrome
Authors: Kogan C. S. , Boutet I. , Cornish K. , Graham G. E. , Berry‐Kravis E. , Drouin A. , Milgram N. W.
Published: November 2008

Background  Standardised neuropsychological and cognitive measures present some limitations in their applicability and generalisability to individuals with intellectual disability (ID). Alternative approaches to defining the cognitive signatures of various forms of ID are needed to advance our understanding of the profiles of strengths and weaknesses as well as the affected brain areas.

Aim  To evaluate the utility and feasibility of six non‐verbal comparative neuropsychological (CN) tasks administered in a modified version of the Wisconsin General Test Apparatus (WGTA) to confirm and extend our knowledge of unique cognitive signatures of Fragile X syndrome (FXS) and Down syndrome (DS).

Method  A test battery of CN tasks adapted from the animal literature was administered in a modified WGTA. Tasks were selected that have established or emerging brain–behaviour relationships in the domains of visual‐perceptual, visual‐spatial, working memory and inhibition.

Results  Despite the fact that these tasks revealed cognitive signatures for the two ID groups, only some hypotheses were supported. Results suggest that whereas individuals with DS were relatively impaired on visual‐perceptual and visual‐spatial reversal learning tasks they showed strengths in egocentric spatial learning and object discrimination tasks. Individuals with FXS were relatively impaired on object discrimination learning and reversal tasks, which was attributable to side preferences. In contrast, these same individuals exhibited strengths in egocentric spatial learning and reversal tasks as well as on an object recognition memory task. Both ID groups demonstrated relatively poor performance for a visual‐spatial working memory task.

Conclusion  Performance on the modified WGTA tasks differentiated cognitive signatures between two of the most common forms of ID. Results are discussed in the context of the literature on the cognitive and neurobiological features of FXS and DS.
Sympathetic and cardiovascular activity during cataplexy in narcolepsy
Authors: DONADIO VINCENZO , PLAZZI GIUSEPPE , VANDI STEFANO , FRANCESCHINI CHRISTIAN , KARLSSON TOMAS , MONTAGNA PASQUALE , VETRUGNO ROBERTO , BUGIARDINI ENRICO , MIGNOT EMMANUEL , LIGUORI ROCCO
Published: November 2008

Autonomic nervous system activity changes have been described during cataplexy as playing a role in triggering it. To confirm these previous findings, we investigated the time course of sympathetic and cardiovascular activities during cataplexy. We made for the first time microneurographic recordings of 10 cataplectic episodes in three patients with hypocretin‐deficient narcolepsy. During microneurography, muscle sympathetic nerve activity (MSNA) was recorded simultaneously with heart rate (HR), respiratory movements, arterial finger blood pressure (BP), electroencephalography, electro‐oculogram and superficial electromyogram. Results showed no significant autonomic changes before the onset of the cataplectic episodes. Cataplexy was associated with a significant increase in MSNA and BP compared with baseline, whereas HR was markedly decreased. An irregular breathing pattern mainly characterized by apnea typically occurred during the attacks. In conclusion, our findings did not show significant changes in autonomic activity prior to cataplexy onset, ruling out a triggering role of the autonomic system. However, cataplexy was associated with co‐activation of sympathetic and parasympathetic autonomic systems, a pattern reminiscent of that reported during the vigilance reaction in animals.
Heterotrimeric G proteins‐mediated resistance to necrotrophic pathogens includes mechanisms independent of salicylic acid‐, jasmonic acid/ethylene‐ and abscisic acid‐mediated defense signaling
Authors: Trusov Yuri , Sewelam Nasser , Rookes James Edward , Kunkel Matt , Nowak Ekaterina , Schenk Peer Martin , Botella José Ramón
Published: November 2008

Heterotrimeric G proteins are involved in the defense response against necrotrophic fungi in Arabidopsis. In order to elucidate the resistance mechanisms involving heterotrimeric G proteins, we analyzed the effects of the Gβ (subunit deficiency in the mutant agb1‐2 on pathogenesis‐related gene expression, as well as the genetic interaction between agb1‐2 and a number of mutants of established defense pathways. Gβ‐mediated signaling suppresses the induction of salicylic acid (SA)‐, jasmonic acid (JA)‐, ethylene (ET)‐ and abscisic acid (ABA)‐dependent genes during the initial phase of the infection with Fusarium oxysporum (up to 48 h after inoculation). However, at a later phase it enhances JA/ET‐dependent genes such as PDF1.2 and PR4. Quantification of the Fusarium wilt symptoms revealed that Gβ‐ and SA‐deficient mutants were more susceptible than wild‐type plants, whereas JA‐ and ET‐insensitive and ABA‐deficient mutants demonstrated various levels of resistance. Analysis of the double mutants showed that the Gβ‐mediated resistance to F. oxysporum and Alternaria brassicicola was mostly independent of all of the previously mentioned pathways. However, the progressive decay of agb1‐2 mutants was compensated by coi1‐21 and jin1‐9 mutations, suggesting that at this stage of F. oxysporum infection Gβ acts upstream of COI1 and ATMYC2 in JA signaling.
Gene silencing by artificial microRNAs in Chlamydomonas
Authors: Zhao Tao , Wang Wei , Bai Xue , Qi Yijun
Published: November 2008

Chlamydomonas reinhardtii is a unicellular green alga. It is a model system for studying functions of the chloroplast, basal body and flagella. The completion of the Chlamydomonas genome sequence makes it possible to use reverse genetic approaches in this organism. Chlamydomonas contains a set of endogenous microRNAs (miRNAs) that down‐regulate their target gene expression through mRNA cleavage. Here we developed an artificial miRNA‐based strategy to knock down gene expression in Chlamydomonas. Using an endogenous Chlamydomonas miRNA precursor as the backbone, we constructed two artificial miRNAs (amiRNAs) targeting the MAA7 and RBCS1/2 genes, respectively. When overexpressed, these two amiRNAs could cleave their respective targets precisely at the predicted sites, resulting in greatly decreased accumulation of MAA7 and RBCS1/2 transcripts and expected mutant phenotypes. We further showed that the two amiRNAs could be produced simultaneously from a dimeric amiRNA precursor. We anticipate that the amiRNA technology developed in this study will be very useful in assessing the functions of individual genes and in genome‐wide approaches.
Conserved intragenic elements were critical for the evolution of the floral C‐function
Authors: Causier Barry , Bradley Desmond , Cook Holly , Davies Brendan
Published: November 2008

The floral C‐function, which specifies stamen and carpel development, played a pivotal role in the evolution of flowers. An important aspect of this was the establishment of mechanisms regulating the temporal and spatial expression domain of the C‐function genes. Transcription of the Arabidopsis C‐function gene AGAMOUS (AG) is tightly controlled by factors that interact with cis‐elements within its large second intron. Little is known about the regulatory role of intragenic elements in C‐function genes from species other than Arabidopsis. We show that a binding site for the LEAFY (LFY) transcription factor, present in the AG intron, is conserved in the introns of diverse C‐function genes and is positioned close to other conserved motifs. Using an in planta mutagenesis approach, we targeted evolutionarily conserved sequences in the intron of the Antirrhinum PLENA (PLE) gene to establish whether they regulate PLE expression. Small sequence deletions resulted in a novel class of heterochronic C‐function mutants with delayed onset of PLE expression and loss of stamen identity. These phenotypes differ significantly from weak C‐function mutant alleles in Antirrhinum and Arabidopsis. Our findings demonstrate that the PLE intron contains regulatory cis‐elements, including a LFY‐binding site, critical for establishing the correct C‐function expression domain. We show that the LFY site, and other conserved intron elements, pre‐date the divergence of the monocot and dicot lineages, suggesting that they were a determinant in the evolution of the C‐function, and propose a threshold model to explain phenotypic divergence observed between C‐function mutants.
Insecticide, sugar, and diet effects on feeding and mortality in Rhagoletis indifferens (Dipt., Tephritidae)
Authors: Yee W. L.
Published: November 2008

The effects of spinosad bait and various insecticides, the presence of sugar in insecticides, and diet on feeding responses and mortality in western cherry fruit fly, Rhagoletis indifferens Curran (Dipt., Tephritidae), were determined. Numbers of feeding events on insecticides with sugar were greater than on insecticides alone, but there was only a small effect of diet on feeding responses to insecticides with sugar. Feeding durations on imidacloprid, thiamethoxam and acetamiprid with sugar were shorter than on sugar water and spinosad bait, as the neonicotinoids paralysed flies quickly. Flies that fed on sugar only (nitrogen‐starved) suffered higher mortalities when exposed to spinosad, thiamethoxam and azinphos‐methyl than to imidacloprid, acetamiprid and indoxacarb, and mortality in between these two groups of treatments when exposed to spinosad bait. Mortalities were greater when sugar was added to insecticides, and were higher in nitrogen‐starved than fully‐fed (yeast extract + sugar fed) flies. Flies that fed once on thiamethoxam were killed more quickly than those that fed once on spinosad bait and spinosad. Results suggest that thiamethoxam is comparable to spinosad in its effects on mortality, and that using it with sugar in bait may also have similar results as using spinosad bait or spinosad. One benefit of using thiamethoxam with sugar may be that it kills flies more quickly, before they can oviposit, than spinosad bait, although whether a fly will feed on it may depend on how much sugar or nitrogenous food it has eaten.
Germany/Australia Index of Sperm Sex Sortability in Elephants and Rhinoceros
Authors: Behr B , Rath D , Hildebrandt TB , Goeritz F , Blottner S , Portas TJ , Bryant BR , Sieg B , Knieriem A , Graaf SP de , Maxwell WMC , Hermes R
Published: November 2008

Contents

Flow cytometric sexing of spermatozoa followed by application in artificial insemination or in vitro fertilization provides a unique opportunity to predetermine the sex of offspring and might enhance the conservation management of endangered species in captivity such as the elephant and rhinoceros. To obtain an indication of the sortability of spermatozoa from these species, the relative DNA differences between X and Y chromosome bearing spermatozoa (fresh, frozen thawed, epididymal) from three rhinoceros species [white (Ceratotherium simum), black (Diceros bicornis), Indian (Rhinoceros unicornis)] and both elephant species, the Asian and the African elephant (Elephas maximus, Loxodonta Africana), were determined through separation of spermatozoa into X and Y chromosome bearing populations, using a modified high speed flow cytometer. The head profile areas of spermatozoa from all five species were measured using light microscopy. By multiplying the relative DNA differences and the head profile areas, the sperm sorting indices were calculated to be 47, 48 and 51 for white, black and Indian rhinoceros respectively. The calculated sorting index for the Asian elephant was 66. In the African elephant, we determined the highest sorting index of 76. These results indicate the practicability of flow cytometric sex sorting of spermatozoa from the tested rhinoceros species and both elephant species. The lower sorting indices in rhinos indicate that sex sorting of spermatozoa from the rhinoceros will be more challenging than in elephants.
Identification of Sperm Subpopulations in Stallion Ejaculates: Changes after Cryopreservation and Comparison with Traditional Statistics
Authors: Ortega‐Ferrusola C , Macías García B , Suárez Rama V , Gallardo‐Bolaños JM , González‐Fernández L , Tapia JA , Rodríguez‐Martinez H , Peña FJ
Published: November 2008

Contents

In an attempt to improve the information obtained after computer‐assisted sperm analysis (CASA), data from five stallions (three ejaculates from each) were analysed before (fresh, extended semen) and after cryopreservation using traditional statistics as well as a cluster analysis. The data matrix consisted of 13 987 observations of individual spermatozoa for fresh, extended semen, and 8305 for frozen–thawed samples. As expected, freezing and thawing resulted in a marked decrease of CASA‐derived variables of sperm kinematics. All sperm velocities were significantly lower in frozen–thawed samples than in samples before cooling. Using sperm velocities, six sperm subpopulations were identified in fresh semen (S1–S6). As such, subpopulations S1 and S2 were characterized by low sperm velocities, subpopulations S3 and S4 corresponded to spermatozoa depicting medium speed values, and finally, subpopulations S5 and S6 were those depicting the highest velocities. After freezing and thawing, four sperm subpopulations were identified, listed as nr FT1 to FT4. While subpopulations FT1–FT3 were characterized by low sperm velocities, and thus corresponded speed‐wise to those listed as S1–S4 for fresh, extended semen, the one called number FT4 in frozen semen was characterized by high velocities, of the same range as that of the subpopulations S5 and S6 for fresh spermatozoa. The sperm subpopulation structure varied among stallions, but the cluster analysis hereby assayed was able to provide valuable information about the freezability of the samples that the customary statistics did not reveal.
Factors Affecting Dystocia in Cattle
Authors: Zaborski D , Grzesiak W , Szatkowska I , Dybus A , Muszynska M , Jedrzejczak M
Published: November 2008

Contents

The paper reviews the various factors affecting dystocia in cattle. It is based mainly on the recent studies found in the literature of the subject but refers occasionally to some older papers as well. The factors are grouped into four main categories: direct factors, phenotypic factors related to calf and cow, non‐genetic and genetic factors. The first group includes malpresentations and uterine torsion. The second one includes: calf birth weight, multiple calvings, perinatal mortality, cow pelvic area, cow body weight and body condition at calving, gestation length. The non‐genetic factors are: cow age and parity, year and season of calving, place of calving, maintenance practises, disorders, calf sex and nutrition. Other non‐genetic factors are the level of hormones in the periparturient period, in vitro production of embryos and embryo cloning. Finally, the genotypes of cow, bull and calf, inbreeding, muscular hypertrophy, selection and quantitative trait loci form the fourth group of genetic factors.
Development of Procedures for Sex‐sorting Frozen–Thawed Bovine Spermatozoa
Authors: Underwood SL , Bathgate R , Maxwell WMC , Evans G
Published: November 2008

Contents

Dairy bull sperm may be sex‐sorted, frozen and used to artificially inseminate heifers with acceptable fertility if the herd is well‐managed. One drawback to the technology is that donor bulls must be located within a short distance of the sorting facility in order to collect semen, which limits the number of bulls from which sorted sperm are available. A successful method used to overcome this limitation in sheep is sex‐sorting from frozen–thawed semen and refreezing for artificial insemination. This technique is attractive to the dairy industry, and therefore a series of three experiments was designed to investigate the optimal methods to prepare, sex‐sort and re‐freeze frozen–thawed bovine sperm. Sperm were prepared for sorting by density gradient separation in either PureSperm® or BoviPure™, followed by staining in one of three diluents (Androhep®, Bovine Sheath Fluid + 0.3% BSA or TALP buffer). Sperm were sorted and collected into Test yolk buffer, and frozen in an extender containing 0, 0.25, 0.375 or 0.5% Equex STM Paste. Frozen–thawed sperm were better orientated (p = 0.006) and had fewer damaged membranes (8.7 ± 0.6% vs 19.5 ± 2.4%; p = 0.003) after centrifugation in PureSperm® rather than BoviPure™ gradients. Sperm orientation (p < 0.05) and motility (69.9 ± 3.0 vs 55.6 ± 4.0; p < 0.001) were highest after staining in Androhep® rather than in TALP buffer. Sperm were more motile (58.2 ± 4.7 vs 38.7 ± 3.5; p < 0.001) and had better acrosome integrity (74.3 ± 2.9 vs 66.8 ± 2.0; p < 0.001) after freezing in an extender containing 0.375% Equex STM Paste than in extender without Equex. Hence, a protocol has been developed to allow frozen–thawed bull sperm to be sex‐sorted with high resolution between the sexes, then re‐frozen and thawed with retention of motility and acrosome integrity.
Influence of Apoptosis in Bovine Embryo’s Development
Authors: Antunes G , Chaveiro A , Santos P , Marques A , Jin HS , Moreira da Silva F
Published: November 2008

Contents

The correlation between apoptosis and early bovine embryonic loss is still not fully elucidated. In the present study, the relationship between the arrest of bovine embryos at the different stages of development and apoptosis was evaluated. We used embryos 7 days after in vitro maturation and fertilization, and morphologic and biochemical apoptotic analyses were performed by using a phase contrast microscope and by the terminal transferase dUTP nick end‐labelling respectively. For the statistic, the apoptotic cell ratio (ACR) was determined as the percentage of apoptotic cells per embryo. To evaluate the relation between ACR and fragmentation pattern, embryos were divided into five groups, groups I–V. To assess the relation between ACR and cytoplasmatic fragmentation, embryos were divided into three groups, according to the fragmentation percentage (<5%; 5–15% and >15%). Of the total 139 embryos included, 65 arrested at 2–8 cells; 14 arrested at 9–16 cells; 18 compacted morula and 42 were non‐arrested blastocysts. The average number of embryonic fragmentation at different stages of the development, 2–8 cells, 9–16 cells, compacted morula and blastocyst, was 16.0 ± 1.5, 28.7 ± 4.4, 4.4 ± 2.4 and 1 ± 0.3 respectively. The embryos at the stage of arrested 9–16 cells and compacted morula had higher ACR than those at the blastocyst stage, excluding the stage of 2–8 cells (the genome is not yet active). The correlation detected between embryonic development and ACR was 0.92 (p < 0.01). It was observed that embryos possessing high fragmentation showed the higher ACR value (r = 0.98, p < 0.05). Comparing the results between fragmentation percentage and ACR, it was observed that the embryos with higher percentage of fragmentation corresponded to higher ACR (r = 0.97, p < 0.01). These results clearly demonstrated that bovine embryonic arrest at different stages of development is correlated with the apoptotic mechanisms.