bluuebunny/crossref_metadata_2025 huggingface dataset:
First Report of Anthracnose Caused by Colletotrichum acutatum on Mung Bean Sprouts in Taiwan
Authors: Shen Y. M. , Liu H. L. , Chang S. T. , Chao C. H.
Published: December 2009

Mung bean sprouts (Vigna radiata (L.) R. Wilczek) are a commonly consumed vegetable in Asian countries. Anthracnose lesions on mung bean sprouts (cv. You-lu-dou from Mayamar) were found in an indoor sprouting facility in Taichung County, Taiwan in May of 2009. The incidence of disease exceeded 90% in some lots. Infected hypocotyls had smooth, diamond-shaped to fusiform brown spots, which became further depressed and enlarged with age. A fungus was isolated on potato dextrose agar (PDA) from symptomatic hypocotyls after surface sterilization in 0.6% NaOCl. Fungal colonies were initially salmon to orange in color and became greenish gray on the surface within a week. Setae were not produced in acervuli that developed on PDA. Conidia in the acervuli were one-celled, cylindrical, and hyaline with an average length and width of 14.8 (7.0 to 23.7) × 4.9 (3.3 to 6.7) μm (n = 53). The internal transcribed spacer (ITS) region was amplified with primers ITS1 and ITS4 (4). The sequence was deposited in GenBank (Accession No. GQ889269). The sequence was 99% identical to that of ATCC 56816 strain of Glomerella acutata (Guerber & J. C. Correll), the teleomorph of Colletotrichum acutatum (J. H. Simmonds) over a 522-bp alignment. Thus, the mung bean pathogen was identified as C. acutatum based on morphological (1) and molecular characters. Pathogenicity of the strain was determined by inoculating mung bean seeds from I-Mei Foods Co. (imported from Australia). The seeds were disinfested in 0.6% NaOCl for 10 min, rinsed with sterile distilled water twice, and immersed in a conidial suspension (1.5 × 104 conidia/ml) of C. acutatum for 10 min. Fifteen inoculated seeds were placed on moistened paper towels, distributed into three flasks, and stored in the dark at 32°C. Symptoms similar to those observed on the original sprouts appeared 3 days later on the hypocotyls of the seedlings and all seedlings were infected by day five. Conidia of C. acutatum were produced on all lesions and colonies of C. acutatum were recovered from symptomatic tissues, fulfilling Koch's postulates. Controls (seed immersed in water) remained symptomless. The pathogenicity test was repeated with similar results. The pathogen has been recorded on mung bean sprouts in Korea (2) and on other Vigna spp. in India (3). To our knowledge, this is the first report of sprout rot of mung bean caused by C. acutatum in Taiwan.

References: (1) B. J. Dyko and J. E. M. Mordue. No. 630 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1979. (2) D. K. Kim et al. Plant Pathol. J. 19:203, 2003. (3) K. P. R. Prasanna. Seed Sci. Technol. 13:821, 1985. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990.
First Report of Limber Pine Dwarf Mistletoe (Arceuthobium cyanocarpum) on Sugar Pine (Pinus lambertiana) from California
Authors: Mathiasen R. , Daugherty C.
Published: December 2009

Limber pine dwarf mistletoe (Arceuthobium cyanocarpum (A. Nelson ex Rydberg) Coulter & Nelson; Viscaceae) severely parasitizes limber pine (Pinus flexilis James) and several other white pines, including western white pine (P. monticola Dougl. ex D. Don) and whitebark pine (P. albicaulis Engelm.), over an extensive geographic range in the western United States (1). However, limber pine dwarf mistletoe has not been previously reported on sugar pine (P. lambertiana Dougl.), another white pine found within the range of limber pine dwarf mistletoe (1). In August 2009, we found a sugar pine infected with limber pine dwarf mistletoe approximately 0.8 km northeast of Tahquitz Peak in the San Jacinto Mountains, California (33°45′24′′N, 116°40′24′′W; elevation 2,640 m). The infected sugar pine was 13 inches (33 cm) in diameter and had 13 infections on five of its lower branches. Ten of the infections were producing mature male and female mistletoe plants with open flowers or developing fruits, respectively. Two of the infected branches were forming witches' brooms in response to infection by limber pine dwarf mistletoe. The infected sugar pine was growing within 3 m of four limber pines severely infected with limber pine dwarf mistletoe. The male and female plants produced on the infected sugar pine were morphologically identical to those growing on the infected limber pines. Limber pine dwarf mistletoe can be distinguished from sugar pine dwarf mistletoe (A. californicum Hawksw. & Wiens), the principal dwarf mistletoe parasitizing sugar pine in California, by its smaller plants (mean height 3 cm versus 8 cm) and flowering period (August to September versus June to July). In an attempt to determine the relative susceptibility of sugar pine to limber pine dwarf mistletoe, we conducted a survey of the infested limber pine stand. Because there were no additional sugar pines growing in the area, it was impossible to assess the general susceptibility of sugar pine to limber pine dwarf mistletoe, but the production of many mature plants from 10 of the infections on the sugar pine suggests this tree species may be highly susceptible. However, this is currently the only known location where sugar pine co-occurs with limber pine dwarf mistletoe (1), so assessing the susceptibility of sugar pine to this dwarf mistletoe will depend on locating additional sites where they co-occur. It should be noted also that previous surveys in the San Jacinto Mountains failed to detect infection by limber pine dwarf mistletoe on sugar pine (1). Specimens of limber pine dwarf mistletoe on sugar pine were collected and deposited at the Deaver Herbarium (ASC), Northern Arizona University, Flagstaff (Accession No. 92697). To our knowledge, this is the first report of limber pine dwarf mistletoe parasitizing sugar pine.

References: (1) F. G. Hawksworth and D. Wiens. USDA For. Serv. Agric. Handb. 709, 1996.
Linear stability of a solid–vapour interface
Authors: Style Robert W. , Worster M. Grae
Published: December 2009

We investigate a system consisting of a condensed phase in contact with its vapour. We derive similarity solutions for vapour and temperature profiles and calculate the condition for the presence of vapour supersaturation adjacent to the condensed phase. We analyse the linear stability of a solid–vapour interface with varying atmospheric conditions. The instability is qualitatively similar to the Mullins–Sekerka instability in binary alloys but the results highlight the important parameters for the solid/vapour problem. We derive the neutral stability condition and results are applied to frost flowers, which are small hoar-frost-like crystals that grow on sea ice, and to physical vapour deposition. The results are applicable to many problems in the wide field of condensed-phase/vapour systems.
SanG, a transcriptional activator, controls nikkomycin biosynthesis through binding to the sanN–sanO intergenic region in Streptomyces ansochromogenes
Authors: He Xihong , Li Rui , Pan Yuanyuan , Liu Gang , Tan Huarong
Published: December 2009

Streptomyces ansochromogenes SanG is a pathway-specific regulator that mainly controls the transcription of two transcriptional units involved in nikkomycin biosynthesis. SanG consists of three major functional domains: an N-terminal Streptomyces antibiotic regulatory protein (SARP) domain, a central ATPase domain, and a C-terminal half homologous to guanylate cyclases belonging to the LuxR family. SanG was expressed in Escherichia coli as a C-terminally His6-tagged protein. The purified SanG-His6 was shown to be a dimer in solution by dynamic light scattering. An electrophoretic mobility-shift assay showed that the purified SanG protein could bind to the DNA fragment containing the bidirectional sanN–sanO promoter region. The SanG-binding sites within the bidirectional sanN–sanO promoter region were determined by footprinting analysis and identified a consensus-directed repeat sequence 5′-CGGCAAG-3′. SanG showed significant ATPase/GTPase activity in vitro, and addition of ATP/GTP enhanced the affinity of SanG for target DNA, but ATP/GTP hydrolysis was not essential for SanG binding to the target DNA. However, real-time reverse transcription PCR showed that mutation of the ATPase/GTPase domain of SanG significantly decreased the transcriptional level of sanN–I and sanO–V. These results indicated that the ATPase/GTPase activity of SanG modulated the transcriptional activation of SanG target genes during nikkomycin biosynthesis.
Aerobic carboxydotrophy under extremely haloalkaline conditions in Alkalispirillum/Alkalilimnicola strains isolated from soda lakes
Authors: Sorokin Dimitry Yu. , Tourova Tatjana P. , Kovaleva Olga L. , Kuenen J. Gijs , Muyzer Gerard
Published: December 2009

Aerobic enrichments from soda lake sediments with CO as the only substrate resulted in the isolation of five bacterial strains capable of autotrophic growth with CO at extremely high pH and salinity. The strains belonged to theAlkalispirillum/Alkalilimnicolacluster in the Gammaproteobacteria, where the ability to oxidize CO, but not growth with CO, has been demonstrated previously. The growth with CO was possible only at an oxygen concentration below 5 % and CO concentration below 20 % in the gas phase. The isolates were also capable of growth with formate but not with H2. The carboxydotrophic growth occurred within a narrow pH range from 8 to 10.5 (optimum at 9.5) and a broad salt concentration from0.3 to 3.5 M total Na+(optimum at 1.0 M). Cells grown on CO had high respiration activity with CO and formate, while the cells grown on formate actively oxidized formate alone. In CO-grown cells, CO-dehydrogenase (CODH) activity was detectable both in soluble and membrane fractions, while the NAD-independent formate dehydrogenase (FDH) resided solely in membranes. The results of total protein profiling and the failure to detect CODH with conventional primers for thecoxLgene indicated that the CO-oxidizing enzyme in haloalkaliphilic isolates might differ from the classical aerobic CODH complex. A singlecbbLgene encoding the RuBisCO large subunit was detected in all strains, suggesting the presence of the Calvin cycle of inorganic carbon fixation. Overall, these results demonstrated the possibility of aerobic carboxydotrophy under extremely haloalkaline conditions.
The BAM complex subunit BamE (SmpA) is required for membrane integrity, stalk growth and normal levels of outer membrane β-barrel proteins in Caulobacter crescentus
Authors: Ryan Kathleen R. , Taylor James A. , Bowers Lisa M.
Published: December 2009

The outer membrane of Gram-negative bacteria is an essential compartment containing a specific complement of lipids and proteins that constitute a protective, selective permeability barrier. Outer membrane β-barrel proteins are assembled into the membrane by the essential hetero-oligomeric BAM complex, which contains the lipoprotein BamE. We have identified a homologue of BamE, encoded by CC1365, which is located in the outer membrane of the stalked alpha-proteobacterium Caulobacter crescentus. BamE associates with proteins whose homologues in other bacteria are known to participate in outer membrane protein assembly: BamA (CC1915), BamB (CC1653) and BamD (CC1984). Caulobacter cells lacking BamE grow slowly in rich medium and are hypersensitive to anionic detergents, some antibiotics and heat exposure, which suggest that the membrane integrity of the mutant is compromised. Membranes of the ΔbamE mutant have normal amounts of the outer membrane protein RsaF, a TolC homologue, but are deficient in CpaC*, an aggregated form of the outer membrane secretin for type IV pili. ΔbamE membranes also contain greatly reduced amounts of three TonB-dependent receptors that are abundant in wild-type cells. Cells lacking BamE have short stalks and are delayed in stalk outgrowth during the cell cycle. Based on these findings, we propose that Caulobacter BamE participates in the assembly of outer membrane β-barrel proteins, including one or more substrates required for the initiation of stalk biogenesis.
Connecting parts with processes: SubtiWiki and SubtiPathways integrate gene and pathway annotation for Bacillus subtilis
Authors: Lammers Christoph R. , Flórez Lope A. , Schmeisky Arne G. , Roppel Sebastian F. , Mäder Ulrike , Hamoen Leendert , Stülke Jörg
Published: December 2009

Bacillus subtilis is the model organism for a large group of Gram-positive bacteria, the Firmicutes. Several online databases have been established over time to manage its genetic and metabolic information, but they differ greatly in their rate of update and their focus on B. subtilis. Therefore, a European systems biology consortium called for an integrated solution that empowers its users to enrich online content. To meet this goal we created SubtiWiki and SubtiPathways, two complementary online tools for gene and pathway information on B. subtilis 168. SubtiWiki (http://subtiwiki.uni-goettingen.de/) is a scientific wiki for all genes of B. subtilis and their protein or RNA products. Each gene page contains a summary of the most important information; sections on the gene, its product and expression; sections concerning biological materials and laboratories; and a list of references. SubtiWiki has been seeded with key content and can be extended by any researcher after a simple registration, thus keeping it always up to date. As a complement, SubtiPathways (http://subtipathways.uni-goettingen.de/) is an online tool for navigation of the metabolism of B. subtilis and its regulation. Each SubtiPathways diagram presents a metabolic pathway with its participating enzymes, together with the regulatory mechanisms that act on their expression and activity, in an intuitive interface that is based on Google Maps. Together, SubtiWiki and SubtiPathways provide an integrated view of the processes that make up B. subtilis and its components, making it the most comprehensive web resource for B. subtilis researchers.
Biodegradation of the xenobiotic organic disulphide 4,4′-dithiodibutyric acid by Rhodococcus erythropolis strain MI2 and comparison with the microbial utilization of 3,3′-dithiodipropionic acid and 3,3′-thiodipropionic acid
Authors: Wübbeler Jan Hendrik , Bruland Nadine , Wozniczka Milena , Steinbüchel Alexander
Published: December 2009

Application of the non-toxic 3,3′-thiodipropionic acid (TDP) and 3,3′-dithiodipropionic acid (DTDP) as precursors for the microbial production of polythioesters (PTEs), a class of biologically persistent biopolymers containing sulphur in the backbone, was successfully established previously. However, synthesis of PTEs containing 4-mercaptobutyrate (4MB) as building blocks could not be achieved. The very harmful 4MB is not used as a PTE precursor or as the carbon source for growth by any known strain. As a promising alternative, the harmless oxidized disulfide of two molecules of 4MB, 4,4′-dithiodibutyric acid (DTDB), was employed for enrichments of bacterial strains capable of biodegradation. Investigation of novel precursor substrates for PTEs and comparison of respective strains growing on TDP, DTDP and DTDB as sole carbon source was accomplished. A broad variety of bacteria capable of using one of these organic sulphur compounds were isolated and compared. TDP and DTDP were degraded by several strains belonging to different genera, whereas all DTDB-utilizing strains were affiliated to the species Rhodococcus erythropolis. Transposon mutagenesis of R. erythropolis strain MI2 and screening of 7500 resulting mutants yielded three mutants exhibiting impaired growth on DTDB. Physiological studies revealed production of volatile hydrogen sulphide and accumulation of significant amounts of 4MB, 4-oxo-4-sulphanylbutanoic acid and succinic acid in the culture supernatants. Based on this knowledge, a putative pathway for degradation of DTDB was proposed: DTDB could be cleaved into two molecules of 4MB, followed by an oxidation yielding 4-oxo-4-sulphanylbutanoic acid. A putative desulphydrase probably catalyses the abstraction of sulphur, thereby generating succinic acid and hydrogen sulphide.
Epidemiology of cholangiocarcinoma: An update focusing on risk factors
Authors: Shin Hai‐Rim , Oh Jin‐Kyoung , Masuyer Eric , Curado Maria‐Paula , Bouvard Veronique , Fang Yue‐Yi , Wiangnon Surapon , Sripa Banchob , Hong Sung‐Tae
Published: December 2009

(Cancer Sci2010; 101: 579–585)

Cholangiocarcinoma is relatively rare, but high incidence rates have been reported in Eastern Asia, especially in Thailand. The etiology of this cancer of the bile ducts appears to be mostly due to specific infectious agents. In 2009, infections with the liver flukes,Clonorchis sinensisorOpistorchis viverrini,were both classified as carcinogenic to humans by the International Agency for Research on Cancer for cholangiocarcinoma. In addition, a possible association between chronic infection with hepatitis B and C viruses and cholangiocarcinoma was also noted. The meta‐analysis of published literature revealed the summary relative risks of infection with liver fluke (bothOpistorchis viverriniandClonorchis sinensis), hepatitis B virus, and hepatitis C virus to be 4.8 (95% confidence interval [95% CI]: 2.8–8.4), 2.6 (95% CI: 1.5–4.6), and 1.8 (95% CI: 1.4–2.4), respectively – liver fluke infection being the strongest risk factor for cholangiocarcinoma. Countries where human liver fluke infection is endemic include China, Korea, Vietnam, Laos, and Cambodia. The number of infected persons withClonorchis sinensisin China has been estimated at 12.5 million with considerable variations among different regions. A significant regional variation inOpistorchis viverriniprevalence was also noted in Thailand (average 9.6% or 6 million people). The implementation of a more intensive preventive and therapeutic program for liver fluke infection may reduce incidence rates of cholangiocarcinoma in endemic areas. Recently, advances have been made in the diagnosis and management of cholangiocarcinoma. Although progress on cholangiocarcinoma prevention and treatment has been steady, more studies related to classification and risk factors will be helpful to develop an advanced strategy to cure and prevent cholangiocarcinoma.
‘Pains of imprisonment’: narratives of the women partners and children of the incarcerated
Authors: Chui Wing Hong
Published: December 2009

This paper focuses on an important line of inquiry in contemporary criminal justice social work research: the collateral consequences of incarceration in the Chinese context. It gives an insight into the impact of incarceration on families from an insider's perspective. An analysis of interviews with 10 partners and 10 children yielded three major themes concerning the impact of imprisonment: the problems associated with the imprisonment of a family member, the coping strategies used and views on the importance of maintaining family ties. It is hoped that this study will give practical guidance regarding what should be done, and how, to reduce the negative consequences brought about by the absence (temporary or permanent) of the imprisoned on their families, including both their partners and vulnerable children.
Administration of a probiotic associated with nasal vaccination with inactivated Lactococcus lactis-PppA induces effective protection against pneumoccocal infection in young mice
Authors: Vintiñi E , Villena J , Alvarez S , Medina M
Published: December 2009

Streptococcus pneumoniae is a serious public health problem, especially in developing countries, where available vaccines are not part of the vaccination calendar. We evaluated different respiratory mucosa immunization protocols that included the nasal administration of Lactococcus lactis-pneumococcal protective protein A (PppA) live, inactivated, and in association with a probiotic (Lc) to young mice. The animals that received Lc by the oral and nasal route presented the highest levels of immunoglobulin (Ig)A and IgG anti-PppA antibodies in bronchoalveolar lavages (BAL) and IgG in serum, which no doubt contributed to the protection against infection. However, only the groups that received the live and inactivated vaccine associated with the oral administration of the probiotic were able to prevent lung colonization by S. pneumoniae serotypes 3 and 14 in a respiratory infection model. This would be related to a preferential stimulation of the T helper type 1 (Th1) cells at local and systemic levels and with a moderate Th2 and Th17 response, shown by the cytokine profile induced in BAL and by the results of the IgG1/IgG2a ratio at local and systemic levels. Nasal immunization with the inactivated recombinant strain associated with oral Lc administration was able to stimulate the specific cellular and humoral immune response and afford protection against the challenge with the two S. pneumoniae serotypes. The results obtained show the probiotic-inactivated vaccine association as a valuable alternative for application to human health, especially in at-risk populations, and are the first report of a safe and effective immunization strategy using an inactivated recombinant strain.
Flk-1+ mesenchymal stem cells aggravate collagen-induced arthritis by up-regulating interleukin-6
Authors: Chen B , Hu J , Liao L , Sun Z , Han Q , Song Z , Zhao R C
Published: December 2009

The immunomodulatory ability of mesenchymal stem cells (MSCs) may be used to develop therapies for autoimmune diseases. Flk-1+ MSCs are a population of MSCs with defined phenotype and their safety has been evaluated in Phase 1 clinical trials. We designed this study to evaluate whether Flk-1+ MSCs conferred a therapeutic effect on collagen-induced arthritis (CIA), an animal model of rheumatic arthritis, and to explore the underlying mechanisms. Flk-1+ MSCs, 1–2 × 106, were injected into CIA mice on either day 0 or day 21. The clinical course of arthritis was monitored. Serum cytokine profile was determined by cytometric bead array kit or enzyme-linked immunosorbent assay. Flk-1+ MSCs and splenocytes co-culture was conducted to explore the underlying mechanisms. Flk-1+ MSCs did not confer therapeutic benefits. Clinical symptom scores and histological evaluation suggested aggravation of arthritis in mice treated with MSCs at day 21. Serum cytokine profile analysis showed marked interleukin (IL)-6 secretion immediately after MSC administration. Results of in vitro culture of splenocytes confirmed that the addition of Flk-1+ MSCs promoted splenocyte proliferation and increased IL-6 and IL-17 secretion. Moreover, splenocyte proliferation was also enhanced in mice treated with MSCs at day 21. Accordingly, MSCs at low concentrations were found to promote lipopolysaccharide-primed splenocytes proliferation in an in vitro co-culture system. We propose that Flk-1+ MSCs aggravate arthritis in CIA model by at least up-regulating secretion of IL-6, which favours Th17 differentiation. When Flk-1+ MSCs are used for patients, we should be cautious about subjects with rheumatoid arthritis.
Thermal ramping rate influences evolutionary potential and species differences for upper thermal limits in Drosophila
Authors: Mitchell Katherine A. , Hoffmann Ary A.
Published: December 2009

1. Thermal tolerance is a key factor limiting insect distributions, but there is limited information on the ability of species to evolve different thermal limits. Recent studies indicate that the experimental protocol influences upper limits, with slower but ecologically relevant rates of warming lowering estimates of tolerance. These effects could also influence genetic and environmental variances that define evolutionary potential.

2. To determine the influence of experimental protocol on estimates of narrow sense heritability (h2n) and other measures of evolutionary potential in Drosophila melanogaster, we conducted family studies on knockdown time when flies were immediately exposed to a high temperature (static) or when temperature was increased to an upper limit (ramping).

3. Estimates of variance components in two populations were obtained using the animal model approach that incorporates information from all relationships among relatives. Coefficients of variation were higher when flies were exposed to a static stress, as were estimates of additive genetic variance and measures of evolvability where genetic variances were standardized by trait means. In contrast, levels of environmental variance were higher under ramping conditions. These effects mean that the narrow sense heritability of thermal resistance was low under slow ramping and did not differ significantly from zero.

4. Differences in thermal limits under both methods were detected among Drosophila species. There was a significant positive relationship between the fast and slow ramping estimates of thermal resistance across species after correction for phylogeny, suggesting similar underlying mechanisms or a history of correlated evolution. However, this result was caused by the strong influence of two taxa.

5. These results suggest that natural populations exhibit lower adaptive potential for upper thermal limits under ramping than estimated from traditional (static) estimates of heat resistance. Even the highly adaptable Drosophila melanogaster appears to have little evolutionary potential to extend its upper thermal range under ramping conditions although species have diverged for this measure.
Analysis of resin tags formation in root canal dentine: a cross sectional study
Authors: Malyk Y. , Kaaden C. , Hickel R. , Ilie N.
Published: December 2009

Aim  To evaluate the length, density and quality of resin tags formed by penetration of various types of adhesive systems into dentinal tubules at various cross section levels of the root canal in correlation to the density of dentinal tubules.

Methodology  Thirty mandibular premolars were instrumented and fibre posts were inserted with three different adhesive systems with and without activator: etch & rinse XP Bond and XP Bond/Self Cure Activator; self‐etch (two‐step) AdheSE and AdheSE/AdheSE DC Activator and self‐etch (one‐step) Hybrid Bond and Hybrid Bond/Hybrid Brushes. The resin tags were evaluated from slices obtained from sections perpendicular to the long axis of the teeth at 3, 6, and 9 mm from the root apex under a Confocal Laser Scanning microscope.

Results  In all groups, lack of continuity of resin tag length, density and quality was observed not only from the cervical to the apical region of each root canal, but also in a mesio‐distal direction to the long axis of the root. Application of etch & rinse adhesive in contrast to the self‐etch adhesives provided the formation of the shorter, but considerably denser, more homogeneous and not interrupted resin tags with similar length. Use of the activator for all types of adhesives significantly increased the completeness (P = 0.014) and continuity (P = 0.024) of resin tags.

Conclusions  None of the investigated adhesives were able to completely infiltrate the dentinal tubules in the entire root canal. Use of the etch & rinse adhesive system and the activators significantly increased the density and the quality of resin tags.
Apexogenesis after initial root canal treatment of an immature maxillary incisor – a case report
Authors: Kvinnsland S. R. , Bårdsen A. , Fristad I.
Published: December 2009

Aim  To present a case where a traumatized, immature tooth still showed capacity for continued root development and apexogenesis after root canal treatment was initiated based on an inaccurate pulpal diagnosis.

Summary  Traumatic dental injuries may result in endodontic complications. Treatment strategies for traumatized, immature teeth should aim at preserving pulp vitality to ensure further root development and tooth maturation. A 9‐year‐old boy, who had suffered a concussion injury to the maxillary anterior teeth, was referred after endodontic treatment was initiated in tooth 21 one week earlier. The tooth had incomplete root length, thin dentinal walls and a wide open apex. The pulp chamber had been accessed, and the pulp canal instrumented to size 100. According to the referral, bleeding from the root made it difficult to fill the root canal with calcium hydroxide. No radiographic signs of apical breakdown were recorded. Based on radiographic and clinical findings, a conservative treatment approach was followed to allow continued root development. Follow‐up with radiographic examination every 3rd month was performed for 15 months. Continued root formation with apical closure was recorded. In the cervical area, a hard tissue barrier developed, which was sealed with white mineral trioxide aggregate (MTA). Bonded composite was used to seal the access cavity. At the final 2 years follow‐up, the tooth showed further root development and was free from symptoms.

Key learning points 

Endodontic treatment of immature teeth may result in a poor long‐term prognosis.

The pulp of immature teeth has a significant repair potential as long as infection is prevented.

Treatment strategies of traumatized, immature permanent teeth should aim at preserving pulp vitality to secure further root development and tooth maturation.

Radiographic interpretation of the periapical area of immature teeth may be confused by the un‐mineralized radiolucent zone surrounding the dental papilla.
Resource partitioning or reproductive isolation: the ecological role of body size differences among closely related species in sympatry
Authors: Okuzaki Yutaka , Takami Yasuoki , Sota Teiji
Published: December 2009

1. Body size differences among coexisting related species are common, but the actual effect of these differences in mitigating interspecific interactions, such as resource competition and reproductive interference, is poorly understood.

2. Local assemblages of the ground beetle genus Carabus (subgenus Ohomopterus) typically consist of two or more species of varying sizes. Through foraging and mating experiments using four Ohomopterus species in parapatry and sympatry, we examined whether interspecific body size differences are effective in partitioning food resources or reducing reproductive interference.

3. Because larval Ohomopterus feed exclusively on earthworms, body size differences may be related to partitioning earthworms of different sizes. However, larvae did not exhibit differences in selectivity or attack success on earthworms of different sizes based on larval body size, indicating little possibility of partitioning food by body size.

4. In contrast, interspecific mating behaviours, such as mate recognition, mounting, and copulation, were hindered when body size differences were large; copulation was frequently accomplished between parapatric species with smaller body size differences.

5. These results suggest that body size differences between species effectively reduce reproductive interference, rather than resource competition. Although body size differences in coexisting closely related species have been considered to function in resource partitioning, they may function primarily in reproductive isolation and thereby facilitate coexistence of species.
The Response of Glucose‐Excited Neurones in the Ventromedial Hypothalamus to Decreased Glucose is Enhanced in a Murine Model of Type 2 Diabetes Mellitus
Authors: Cotero V. E. , Zhang B. B. , Routh V. H.
Published: December 2009

Obesity and type 2 diabetes mellitus (T2DM) are associated with dysfunctional insulin signalling and impaired central glucose sensing. Glucose sensing neurones reside in key areas of the brain involved in glucose and energy homeostasis (e.g. ventromedial hypothalamus; VMH). We have recently shown that insulin attenuates the ability of glucose‐excited (GE) neurones to sense decreased glucose. We hypothesise that this effect of insulin on VMH GE neurones is impaired during T2DM when insulin signalling is dysfunctional. To test our hypotheses, we used whole cell patch clamp recording techniques to evaluate the effects of insulin on VMH GE neurones in brain slices from wild‐type and diabetic (db/db) mice. The effects of decreasing glucose from 2.5 to 0.1 mm on VMH GE neurones were similar in wild‐type and db/db mice. However, decreasing glucose from 2.5 to 0.5 mm decreased the action potential frequency, membrane potential and input resistance of VMH GE neurones to a significantly greater extent in db/db versus wild‐type mice. Furthermore, insulin (5 nm) blunted the effects of decreased glucose in wild‐type, but not db/db mice. These differences in both glucose and insulin sensitivity between wild‐type and db/db mice were completely ameliorated by the insulin sensitiser, Compound 2 (300 nm). These data are consistent with our hypothesis that impaired insulin signalling in T2DM sensitises VMH GE neurones to decreased glucose.
Effects of Photoperiod on Kisspeptin Neuronal Populations of the Ewe Diencephalon in Connection with Reproductive Function
Authors: Chalivoix S. , Bagnolini A. , Caraty A. , Cognié J. , Malpaux B. , Dufourny L.
Published: December 2009

Kisspeptin (Kiss) is a key regulator of reproductive function in both prepubertal and adult mammals. Its expression appears to vary throughout the year in seasonal species. We aimed to determine the impact of a change of photoperiod on the size of Kiss neuronal populations found in the preoptic area (POA) and arcuate nucleus (ARC) of the ewe brain. Using immunocytochemistry, we first examined the proportion of neurones expressing Kiss, using HuC/D as a neuronal marker, at different time‐points after transition from long days (LD; 16 : 8 h light/dark cycle) to short days (SD; 8 : 16 h light/dark cycle). Luteinising hormone (LH) secretion was measured in ovariectomised oestradiol replaced ewes from the month preceding the transition to SD until the sacrifice of the animals at days 0, 45 and 112 from this photoperiodic transition. High LH levels were only observed in animals killed at day 112. The number of Kiss neurones/mm2 doubled in the caudal ARC at day 112. The percentage of neurones showing Kiss immunoreactivity increased significantly in both the POA and ARC in the day 112 group. In a second experiment, ewes kept in LD received an i.c.v. injection of colchicine 20 h before sacrifice. Colchicine treatment increased the number and the percentage of neurones with Kiss in both the POA and caudal ARC. The data obtained suggest that the increase in Kiss neurones detected in the POA and caudal ARC after transition to SD stemmed from an increase in Kiss synthesis. This up‐regulation of Kiss content under the shorter day condition appears to be a late event within the cascade activated by a longer secretion of melatonin, which is a critical factor in switching gonadotrophin‐releasing hormone secretion to a breeding season profile.
A Comparison of Two Repeated Restraint Stress Paradigms on Hypothalamic‐Pituitary‐Adrenal Axis Habituation, Gonadal Status and Central Neuropeptide Expression in Adult Male Rats
Authors: Gray M. , Bingham B. , Viau V.
Published: December 2009

The available evidence continues to illustrate an inhibitory influence of male gonadal activity on the hypothalamic‐pituitary‐adrenal (HPA) axis under acute stress. However, far less is known about how these systems interact during repeated stress. Because HPA output consistently declines across studies examining repeated restraint, the potential mechanisms mediating this habituation are often inferred as being equivalent, even though these studies use a spectrum of restraint durations and exposures. To test this generalisation, as well as to emphasise a potential influence of the male gonadal axis on the process of HPA habituation, we compared the effects of two commonly used paradigms of repeated restraint in the rodent: ten daily episodes of 0.5 h of restraint and five daily episodes of 3 h of restraint. Both paradigms produced comparable declines in adrenocorticotrophic hormone and corticosterone between the first and last day of testing. However, marked differences in testosterone levels, as well as corticotrophin‐releasing hormone (CRH) and arginine vasopressin (AVP) expression, occurred between the two stress groups. Plasma testosterone levels remained relatively higher in animals exposed to 0.5 h of restraint compared to 3 h of restraint, whereas forebrain gonadotrophin‐releasing hormone (GnRH) cell counts increased in both groups. AVP mRNA was increased after 3 h, but not after 0.5 h of repeated restraint, in the medial parvicellular paraventricular nucleus and in the posterior bed nucleus of the stria terminalis (BST), and increased with 0.5 h of repeated restraint in the medial amygdala. CRH mRNA was increased after 3 h, but not after 0.5 h of repeated restraint, in the central amygdala and anterior BST. The data obtained illustrate that, despite comparable declines in HPA responses, the pathways recruited for stress adaptation appear to be distinct between restraint groups. Given the extreme sensitivity of limbic AVP to testosterone, and conversely CRH to circulating glucocorticoids, whether differences in endocrine profiles might explain these neuropeptide differences remains to be seen. Nonetheless, the present study provides several new entry points for testing gonadal influences on stress‐specific HPA habituation.
The impact of epistatic selection on the genomic traces of selection
Authors: OTTO SARAH P. , WHITLOCK MICHAEL C.
Published: December 2009

The rapid accumulation of genomic data has led to an explosion of studies searching for signals of past selection left within DNA sequences. Yet the majority of theoretical studies investigating the traces of selection have assumed a simple form of selection, without interactions among selectively fixed sites. Fitness interactions—‘epistasis’—are commonplace, however, and take on a myriad of forms (Whitlock et al. 1995; Segrèet al. 2005; Phillips 2008). It is thus important to determine how such epistasis would influence selective sweeps. On p. 5018 of this issue, Takahasi (2009) explores the effect of epistasis on genetic variation neighbouring two sites that interact in determining fitness, finding that such epistasis has a dramatic impact on the genetic variability in regions surrounding the interacting sites.
Ribosome reactivation by replacement of damaged proteins
Authors: Pulk Arto , Liiv Aivar , Peil Lauri , Maiväli Ülo , Nierhaus Knud , Remme Jaanus
Published: December 2009

Ribosomal functions are vital for all organisms. Bacterial ribosomes are stable 2.4 MDa particles composed of three RNAs and over 50 different proteins. Accumulating damage to ribosomal RNA or proteins can disturb ribosome functioning. Organisms could benefit from degrading or possibly repairing inactive or partially active ribosomes. Reactivation of chemically damaged ribosomes by a process of protein replacement was studied in vitro. Ribosomes were inactivated by chemical modification of Cys residues. Incubation of modified ribosomes with total ribosomal proteins led to reactivation of translational activity. Intriguingly, ribosomal proteins extracted by LiCl are equally active in the restoration of ribosome function. Incubation of 70S ribosomes with isotopically labelled r‐proteins followed by separation of ribosomes was used to identify exchangeable proteins. A similar set of proteins was found to be exchanged in vivo under stress conditions in the stationary phase. We propose that repair of damaged ribosomes might be an important mechanism for maintaining protein synthesis activity following chemical damage.
Functional specialization and differential regulation of short‐chain carboxylic acid transporters in the pathogen Candida albicans
Authors: Vieira Neide , Casal Margarida , Johansson Björn , MacCallum Donna M. , Brown Alistair J.P. , Paiva Sandra
Published: December 2009

The major fungal pathogen Candida albicans has the metabolic flexibility to assimilate a wide range of nutrients in its human host. Previous studies have suggested that C. albicans can encounter glucose‐poor microenvironments during infection and that the ability to use alternative non‐fermentable carbon sources contributes to its virulence. JEN1 encodes a monocarboxylate transporter in C. albicans and we show that its paralogue, JEN2, encodes a novel dicarboxylate plasma membrane transporter, subjected to glucose repression. A strain deleted in both genes lost the ability to transport lactic, malic and succinic acids by a mediated mechanism and it displayed a growth defect on these substrates. Although no significant morphogenetic or virulence defects were found in the double mutant strain, both JEN1 and JEN2 were strongly induced during infection. Jen1‐GFP (green fluorescent protein) and Jen2‐GFP were upregulated following the phagocytosis of C. albicans cells by neutrophils and macrophages, displaying similar behaviour to an Icl1‐GFP fusion. In the murine model of systemic candidiasis approximately 20–25% of C. albicans cells infecting the kidney expressed Jen1‐GFP and Jen2‐GFP. Our data suggest that Jen1 and Jen2 are expressed in glucose‐poor niches within the host, and that these short‐chain carboxylic acid transporters may be important in the early stages of infection.
Deploying artemether‐lumefantrine with rapid testing in Ethiopian communities: impact on malaria morbidity, mortality and healthcare resources
Authors: Lemma Hailemariam , Byass Peter , Desta Alem , Bosman Andrea , Costanzo Gianfranco , Toma Luigi , Fottrell Edward , Marrast Anne‐Claire , Ambachew Yohannes , Getachew Asefaw , Mulure Nathan , Morrone Aldo , Bianchi Angela , Barnabas Gebre Ab
Published: December 2009

Objective  To assess the impact and feasibility of artemether‐lumefantrine deployment at community level, combined with phased introduction of rapid diagnostic tests (RDTs), on malaria transmission, morbidity, and mortality and health service use in a remote area of Ethiopia.

Methods  Two‐year pilot study in two districts: artemether‐lumefantrine was prescribed after parasitological confirmation of malaria in health facilities in both districts. In the intervention district, artemether‐lumefantrine was also made available through 33 community health workers (CHWs); of these, 50% were equipped with RDTs in the second year.

Results  At health facilities; 54 774 patients in the intervention and 100 535 patients in the control district were treated for malaria. In the intervention district, 75 654 patients were treated for malaria by community health workers. Use of RDTs in Year 2 excluded non‐Plasmodium falciparumin 89.7% of suspected cases. During the peak of malaria transmission in 2005, the crude parasite prevalence was 7.4% (95% CI: 6.1–8.9%) in the intervention district and 20.8% (95% CI: 18.7–23.0%) in the control district. Multivariate modelling indicated no significant difference in risk of all‐cause mortality between the intervention and the control districts [adjusted incidence rate ratio (aIRR) 1.03, 95%CI 0.87–1.21, P = 0.751], but risk of malaria‐specific mortality was lower in the intervention district (aIRR 0.60, 95%CI 0.40–0.90, P = 0.013).

Conclusions  Artemether‐lumefantrine deployment through a community‐based service in a remote rural population reduced malaria transmission, lowered the malaria case burden for health facilities and reduced malaria morbidity and mortality during a 2‐year period which included a major malaria epidemic.
Collaborative study to establish a replacement World Health Organization International Standard for parvovirus B19 DNA nucleic acid amplification technology (NAT)‐based assays
Authors: Baylis S. A. , Chudy M. , Blümel J. , Pisani G. , Candotti D. , José M. , Heath A. B.
Published: December 2009

Background and objectives  The aim of the study was to replace the 1st World Health Organization International Standard for parvovirus B19 DNA for nucleic acid amplification technique (NAT)‐based assays (code 99/800). Two lyophilized preparations (coded 99/800 and 99/802) had been evaluated in the original collaborative study. The present study re‐evaluates these two preparations in terms of potency, stability and encapsidation of virus DNA.

Materials and methods  The 1st International Standard (99/800) and 99/802 were re‐coded as Samples 1 and 2, respectively. The samples were distributed to six laboratories and assayed on four separate occasions. Accelerated thermal degradation samples of the two preparations were examined after storage at 20°C for 7 years. Nuclease treatment was used to investigate the encapsidation of virus DNA.

Results  Data were returned from a total of six different quantitative NAT‐based assays. The results of the present study confirm those of the original, with no significant differences found in estimated international units (IU)/ml for the 1st International Standard (Sample 1 in this study) and the proposed replacement preparation, Sample 2 (99/802). Accelerated thermal degradation studies demonstrate that both samples are very stable, with no loss of potency after storage at 20°C for 7 years. Both lyophilized preparations contained the majority of B19V DNA encapsidated in virions.

Conclusions  On the basis of the data presented in this collaborative study, Sample 2 (code number 99/802) was established as the 2nd International Standard for parvovirus B19 DNA for NAT‐based assays with a potency of 106 IU/ml (500 000 IU/vial).
No Correlation Between Song and Circulating Testosterone Levels During Multiple Broods in the Domesticated Canary (Serinus canaria)
Authors: Voigt Cornelia , Leitner Stefan
Published: December 2009

In anticipation of the breeding season male songbirds of the temperate zones undergo gonadal recrudescence in early spring that lead to elevated circulating testosterone (T) levels, positively correlated with an increase in aggressive and song behaviour. However, besides seasonal changes there are also marked fluctuations of T levels and song production within the breeding season. In many species, T levels and singing activity drop after pairing or after the first clutch is laid. Domesticated canaries (Serinus canaria) are multiple‐brooded with an extended breeding season, and males continue to sing after egg‐laying. So far, studies have mainly focused on the seasonality of T levels and song behaviour whereas the pattern of change throughout the breeding period is unknown. Here, we focused on the first and on the last brood of the breeding season. We measured plasma T levels in males at the different breeding stages and assessed song characteristics of males at both times. T levels fluctuated significantly throughout brood 1, being highest during the nest building stage compared with egg‐laying and feeding of young. No such changes occurred during the last brood. Temporal song characteristics changed between brood 1 and brood 3 with song length being the main contributor to explain these changes. Our data suggest that T mainly plays a role in mate attraction and initial nesting site selection but that elevated levels are not necessary for subsequent breeding attempts. Furthermore, temporal song characteristics are maintained independently of T levels, suggesting a threshold effect. Our results demonstrate behavioural and physiological plasticity of domesticated canaries during the breeding season and are consistent with previous findings in wild songbirds.