bluuebunny/crossref_metadata_2025 huggingface dataset:
Contrasting demographic and genetic estimates of dispersal in the endangered Coahuilan box turtle: a contemporary approach to conservation
Authors: HOWETH JENNIFER G. , MCGAUGH SUZANNE E. , HENDRICKSON DEAN A.
Published: August 2008

The evolutionary viability of an endangered species depends upon gene flow among subpopulations and the degree of habitat patch connectivity. Contrasting population connectivity over ecological and evolutionary timescales may provide novel insight into what maintains genetic diversity within threatened species. We employed this integrative approach to evaluating dispersal in the critically endangered Coahuilan box turtle (Terrapene coahuila) that inhabits isolated wetlands in the desert‐spring ecosystem of Cuatro Ciénegas, Mexico. Recent wetland habitat loss has altered the spatial distribution and connectivity of habitat patches; and we therefore predicted thatT. coahuilawould exhibit limited movement relative to estimates of historic gene flow. To evaluate contemporary dispersal patterns, we employed mark–recapture techniques at both local (wetland complex) and regional (intercomplex) spatial scales. Gene flow estimates were obtained by surveying genetic variation at nine microsatellite loci in seven subpopulations located across the species’ geographical range. The mark–recapture results at the local spatial scale reveal frequent movement among wetlands that was unaffected by interwetland distance. At the regional spatial scale, dispersal events were relatively less frequent between wetland complexes. The complementary analysis of population genetic substructure indicates strong historic gene flow (globalFST = 0.01). However, a relationship of genetic isolation by distance across the geographical range suggests that dispersal limitation exists at the regional scale. Our approach of contrasting direct and indirect estimates of dispersal at multiple spatial scales inT. coahuilaconveys a sustainable evolutionary trajectory of the species pending preservation of threatened wetland habitats and a range‐wide network of corridors.
SreA‐mediated iron regulation in Aspergillus fumigatus
Authors: Schrettl Markus , Kim H. Stanley , Eisendle Martin , Kragl Claudia , Nierman William C. , Heinekamp Thorsten , Werner Ernst R. , Jacobsen Ilse , Illmer Paul , Yi Hyojeong , Brakhage Axel A. , Haas Hubertus
Published: August 2008

Aspergillus fumigatus, the most common airborne fungal pathogen of humans, employs two high‐affinity iron uptake systems: iron uptake mediated by the extracellular siderophore triacetylfusarinine C and reductive iron assimilation. Furthermore, A. fumigatus utilizes two intracellular siderophores, ferricrocin and hydroxyferricrocin, to store iron. Siderophore biosynthesis, which is essential for virulence, is repressed by iron. Here we show that this control is mediated by the GATA factor SreA. During iron‐replete conditions, SreA deficiency partially derepressed synthesis of triacetylfusarinine C and uptake of iron resulting in increased cellular accumulation of both iron and ferricrocin. Genome‐wide DNA microarray analysis identified 49 genes that are repressed by iron in an SreA‐dependent manner. This gene set, termed SreA regulon, includes all known genes involved in iron acquisition, putative novel siderophore biosynthetic genes, and also genes not directly linked to iron metabolism. SreA deficiency also caused upregulation of iron‐dependent and antioxidative pathways, probably due to the increased iron content and iron‐mediated oxidative stress. Consistently, the sreA disruption mutant displayed increased sensitivity to iron, menadion and phleomycin but retained wild‐type virulence in a mouse model. As all detrimental effects of sreA disruption are restricted to iron‐replete conditions these data underscore that A. fumigatus faces iron‐depleted conditions during infection.
Evidence for photic‐zone euxinia in the Early Albian Santana Formation (Araripe Basin, NE Brazil)
Authors: Heimhofer U. , Hesselbo S. P. , Pancost R. D. , Martill D. M. , Hochuli P. A. , Guzzo J. V. P.
Published: August 2008

Organic matter from two black shale units in the Santana Formation, Araripe Basin, NE Brazil, has been studied to investigate the palaeoenvironmental conditions prevailing during formation of this famous fossil lagerstätte. Optical and organic geochemical analyses indicate a predominantly autochthonous source with very limited terrestrial input. Molecular fossils of green sulphur bacteria show that the water column was stratified with euxinic conditions reaching up into the photic zone. Thus, mass‐mortality events of fish may be explained by intrusion of sulphide‐containing waters. The low quantity of material derived from land plants in this interior marine setting is interpreted to reflect a consequence of enhanced marine productivity, resulting in dilution of the continental signal. Different depositional scenarios are proposed to explain our findings including (1) coastal erosion and nutrient leaching during a transgressive event, (2) intrusion of euxinic waters during an early Albian OAE or (3) an estuarine circulation pattern associated with thermohaline stratification.
Ophiolitic trondhjemites: a possible analogue for Hadean felsic 'crust'
Authors: Rollinson Hugh
Published: August 2008

It has been argued that >4.0 Ga detrital zircons preserved in sediments of the Jack Hills, western Australia, preserve evidence for a well‐developed continental crust on the Earth at 4.4–4.5 Ga ago. Here, it is shown that there are geochemical similarities between the Jack Hills zircons and the zircons found in trondhjemites in ophiolite sequences, suggesting that the Earth's first felsic crust may have formed in a manner analogous to modern ophiolitic trondhjemites. The trondhjemites of the Oman ophiolite were formed by the hydrous partial melting of the upper (hornblende) gabbros in the roof‐zone of an axial magma chamber. A similar hydrous melting of a mafic protolith may have operated during the Hadean, to create small volumes of felsic rocks within a dominantly mafic crust, obviating the need to postulate a felsic continental crust at 4.4–4.5 Ga.
The importance of context in malaria diagnosis and treatment decisions ‐ a quantitative analysis of observed clinical encounters in Tanzania
Authors: Chandler Clare I. R. , Chonya Semkini , Boniface Gloria , Juma Kaseem , Reyburn Hugh , Whitty Christopher J. M.
Published: August 2008

Objective  To gain a better understanding of the decision‐making context in the diagnosis of malaria in order to inform behaviour change strategies, using quantitative methods.

Methods  We observed hospital outpatient and inpatient consultations in northeast Tanzania where malaria testing was routinely available, recording potential influences on testing and prescribing decisions. We analysed the effects of variables at patient, clinical context and clinician levels on three key decisions in malaria diagnosis and treatment: decision to test for malaria, presumptive treatment and treatment of test‐negative patients with antimalarials.

Results  Observation of 2082 consultations took place during 120 clinics (different shifts on different days and in different departments) with 34 clinicians. Malaria tests were requested for 16.9% of patients. This decision was driven primarily by clinical symptoms. Of patients not tested for malaria, 36.0% were prescribed antimalarials, this decision being associated with both clinical and non‐clinical factors. In outpatients fever was a strong predictor of presumptive treatment [adjusted odds ratio (AOR): 45.9, 95% CI: 30–73], in inpatients this was less so (AOR: 2.7, 95% CI: 0.98–7.7). Outpatient clinicians who were working alone or who had attended <2 in‐service training sessions in the past year were more likely to prescribe antimalarials presumptively. The decision to prescribe antimalarials without also prescribing antibiotic treatment to 22.8% patients who tested negative for malaria was not driven by clinical symptoms but was associated with age over 5 years, lower patient load and male sex of clinician.

Conclusions  Non‐clinical factors are important in the overdiagnosis of malaria. Strategies to target antimalarials and antibiotics better need to use methods that address the context of clinical decision making in addition to the dissemination of conventional clinical algorithms.
Nested PCRs and sequencing of nuclear ITS‐rDNA fragments detect three Leishmania species of gerbils in sandflies from Iranian foci of zoonotic cutaneous leishmaniasis
Authors: Parvizi P. , Ready P. D.
Published: August 2008

Objective  To identify and understand the natural transmission cycles of Leishmania in Iranian sandflies.

Method  Nested PCR protocols were developed to amplify two regions of the ribosomal RNA amplicon of Leishmania (ITS1‐5.8S rRNA gene, and a microsatellite DNA region of ITS2), which were species‐specific by DNA sequence or fragment size.

Results  The PCR assays detected in Iranian sandflies not only Leishmania major but also for the first time L. turanica and L. gerbilli sensu lato, two other parasites of the great gerbil. All three parasites were found in the northeast and centre of Iran, in two foci of rural Zoonotic Cutaneous Leishmaniasis (ZCL) caused by L. major. Fifty infections were detected in common sandfly species: at least six geographically differentiated haplotypes of L. major in four to five sandfly species; one strain of L. gerbilli s.l. in five to six sandfly species and one strain of L. turanica in one sandfly species.

Conclusion  Past conclusions about the transmission cycles of L. major in Iran should be treated with caution. Careful molecular eco‐epidemiological investigations are essential for modelling the roles of different sandfly species in maintaining and spreading ZCL foci. Even if non‐pathogenic to humans, frequent inoculations of L. turanica by sandflies might alter the efficacy of vaccines against L. major. Phlebotomus papatasi is probably the key vector in many ZCL foci because of its abundance and high infection rates with both L. major and L. turanica.
Envenoming after carpet viper (Echis ocellatus) bite during pregnancy: timely use of effective antivenom improves maternal and foetal outcomes
Authors: Habib A. G. , Abubakar S. B. , Abubakar I. S. , Larnyang S. , Durfa N. , Nasidi A. , Yusuf P. O. , Garnvwa J. , Theakston R. D. G. , Salako L. , Warrell D. A. ,   for the EchiTab Study Group (Nigeria & UK)
Published: August 2008

The report describes successful management of 10 women in 2nd and 3rd pregnancy trimesters with EchiTab IgG antivenom after carpet viper (Echis ocellatus) envenoming. All women survived but foetal loss in a victim with delayed presentation and a case of mild hypersensitivity reaction were recorded. Excellent outcomes can be achieved in rural and semi‐nomadic populations without specialized care and immediate access and provision of effective antivenoms is paramount in curtailing snakebite maternal morbidity, mortality and foetal loss.
Introduction of Aedes albopictus in Gabon: what consequences for dengue and chikungunya transmission?
Authors: Vazeille Marie , Moutailler Sara , Pages Frédéric , Jarjaval Fanny , Failloux Anna‐Bella
Published: August 2008

The 2007 outbreak of chikungunya in Gabon has indicated the potential of this disease to spread beyond its usual range ensuing from the expansion of the mosquito Aedes albopictus. A few cases of dengue (DEN) infection were also detected. Because little is known about the potential for Gabonese mosquito species to transmit both chikungunya and DEN viruses (DENV), we conducted studies to determine the susceptibility of Ae. albopictus and Aedes aegypti collected in Libreville to both viruses by experimental infections. Disseminated infection rates were high for Ae. albopictus infected with chikungunya virus (CHIKV) (66.7–86%) and low with DENV (13–21.4%). Moreover, Ae. aegypti sp. formosus was a less efficient vector of CHIKV than Ae. albopictus. The recent introduction and dissemination of chikungunya associated with the invasion of Ae. albopictus in Africa illustrates the potential for CHIKV to spread to other parts of the world.
Dynamics of dengue epidemics in urban contexts
Authors: Pongsumpun P. , Garcia Lopez D. , Favier C. , Torres L. , Llosa J. , Dubois M. A.
Published: August 2008

Dengue, similar to other arboviral diseases, exhibits complex spatiotemporal dynamics. Even at town or village level, individual‐based spatially explicit models are required to correctly reproduce epidemic curves. This makes modelling at the regional level (province, country or continent) very difficult and cumbersome. We propose here a first step to build a hierarchized model by constructing a simple analytical expression which reproduces the model output from macroscopic parameters describing each ‘village’. It also turns out to be a good approximation of real urban epidermic outbreaks. Subsequently, a regional model could be built by coupling these equations on a lattice.
Vertically acquired paediatric HIV infection: the challenges of providing comprehensive packages of care in resource‐limited settings
Authors: Little K. E. , Bland R. M. , Newell M. L.
Published: August 2008

The successes achieved in paediatric disease management in well‐resourced countries in recent years highlight the vast divide between the care options, and ultimately survival, between developed and developing areas of the world. Using an extensive literature review, we quantify recent achievements in terms of improved survival and quality of life, and examine current evidence of the effects of treatment on the survival and morbidity of HIV‐infected children in developing countries. When provided with the same care as their counterparts in developed countries, children in developing countries show similar improvements in survival and general health, with 1‐year survival rates exceeding 90% in many African settings. Despite the challenges of providing comprehensive packages of care in resource‐limited settings, there is an urgent need to scale up prevention and treatment of HIV infections in children, focussing on strengthening Prevention of Mother‐to‐Child Transmission programmes in order to reduce the numbers of infants who are infected in addition to reducing morbidity and mortality among their mothers.
Causes of deaths using verbal autopsy among adolescents and adults in rural western Kenya
Authors: Van Eijk A. M. , Adazu K. , Ofware P. , Vulule J. , Hamel M. , Slutsker L.
Published: August 2008

Objective  To establish causes and patterns of deaths among adolescents and adults (age >11 years) using verbal autopsy (VA) in a rural area of western Kenya where malaria and HIV are common.

Methods  Village reporters reported all deaths in Asembo and Gem (population 135 000), an area under routine demographic surveillance. After an interval of ≥1 month, a trained interviewer used a structured questionnaire to ask the caretaker about signs and symptoms that preceded death. Three clinical officers independently reviewed the interviews and assigned two unranked causes of death; a common cause was designated as the cause of death.

Results  In 2003, 1816 deaths were reported from residents; 48% were male and 72% were between 20 and 64 years of age. Most residents (97%) were ill before death, with 60% of illnesses lasting more than 2 months; 87% died at home. Care was sought by 96%; a health facility was the most common source, visited by 73%. For 1759 persons (97%), a common cause of death was designated. Overall, 74% of deaths were attributed to infectious causes. HIV (32%) and tuberculosis (TB) (16%) were the most frequent, followed by malaria, respiratory infections, anaemia and diarrhoeal disease (approximately 6% each). Death in a health facility was associated with young age, higher education, higher SES, a non‐infectious disease cause and a shorter duration of illness.

Conclusion  In this area, the majority of adult and adolescent deaths were attributed to potentially preventable infectious diseases. Deaths in health facilities were not representative of deaths in the community. Programmes to prevent HIV and TB infection and to decrease mortality have started. Their impact can be evaluated against this baseline information.
TB, HIV‐associated TB and multidrug‐resistant TB on Thailand’s border with Myanmar, 2006–2007
Authors: Hemhongsa Patjuban , Tasaneeyapan Theerawit , Swaddiwudhipong Witaya , Danyuttapolchai Junya , Pisuttakoon Kanoknart , Rienthong Somsak , McCarthy Kimberly , Varma Melissa J. , Whitmore Jacqueline , Varma Jay K.
Published: August 2008

Objective  To measure the burden and improve management of tuberculosis (TB), HIV‐associated TB and MDR TB in Tak Province, Thailand, which borders Myanmar.

Methods  From September 2006 to August 2007, we collected uniform data about TB cases and enhanced human immunodeficiency virus (HIV) counselling and testing. We provided mycobacterial culture and drug‐susceptibility testing in public or non‐governmental organization facilities. Patients were classified by nationality and, for non‐Thais, by migration status.

Results  Of 1662 TB cases in the 12‐month period, 1087 (65%) occurred in non‐Thais. Of non‐Thais, 415 (38%) lived in Myanmar but crossed the border for healthcare. HIV infection was diagnosed in 18% of Thais compared with 12% of non‐Thais (P < 0.01); HIV status was unknown for 22% of Thais and 27% of non‐Thais (P = 0.02). Overall, multidrug‐resistant (MDR) TB was diagnosed in 27 patients, 19 (70%) in non‐Thais. Among TB cases never previously treated for TB, no MDR cases were diagnosed in Thais or in Myanmar refugees, but six cases were diagnosed in migrants from Myanmar.

Conclusions  In Thailand, TB, HIV‐associated TB and MDR TB in migrants from Myanmar are important public health problems; they need to be resolved in both the countries.
Recurrent costs of HIV/AIDS‐related health services in Rwanda: implications for financing
Authors: Quentin Wilm , König Hans‐Helmut , Schmidt Jean‐Olivier , Kalk Andreas
Published: August 2008

Objective  To estimate recurrent costs per patient and costs for a national HIV/AIDS treatment programme model in Rwanda.

Methods  A national HIV/AIDS treatment programme model was developed. Unit costs were estimated so as to reflect necessary service consumption of people living with HIV/AIDS (PLWHA). Two scenarios were calculated: (1) for patients/clients in the year 2006 and (2) for potential increases of patients/clients. A sensitivity analysis was conducted to test the robustness of results.

Results  Average yearly treatment costs were estimated to amount to 504 US$ per patient on antiretroviral therapy (ART) and to 91 US$ for non‐ART patients. Costs for the Rwandan HIV/AIDS treatment programme were estimated to lie between 20.9 and 27.1 million US$ depending on the scenario. ART required 9.6 to 11.1 million US$ or 41–46% of national programme costs. Treatment for opportunistic infections and other pathologies consumed 7.1 to 9.3 million US$ or 34% of total costs.

Conclusion  Health Care in general and ART more specifically is unaffordable for the vast majority of Rwandan PLWHA. Adequate resources need to be provided not only for ART but also to assure treatment of opportunistic infections and other pathologies. While risk‐pooling may play a limited role in the national response to HIV/AIDS, considering the general level of poverty of the Rwandan population, no appreciable alternative to continued donor funding exists for the foreseeable future.
Tryptophan catabolites regulate mucosal sensitization to ovalbumin in respiratory airways
Authors: Odemuyiwa S. O. , Ebeling C. , Duta V. , Abel M. , Puttagunta L. , Cravetchi O. , Majaesic C. , Vliagoftis H. , Moqbel R.
Published: August 2008

Background:  Indoleamine 2,3 dioxygenase (IDO), the rate‐limiting enzyme in tryptophan catabolism, is important in generating tolerance at the foetal–maternal interface. Studies using 1‐methyl‐tryptophan (1‐MT), the specific inhibitor of IDO, showed that this enzyme is important in interferon‐gamma (IFN‐γ)‐dependent inhibition of allergic inflammation in the respiratory airway during immunotherapy.

Aims of study:  We investigated the role of IDO in the development of allergic sensitization, leading to allergic inflammation and airway hyper‐responsiveness (AHR).

Methods:  We used a mouse model to generate mucosal tolerance to lipopolysaccharide‐free ovalbumin (OVA) following repeated intranasal inoculation of OVA over a 3‐day period. We tested the successful induction of tolerance by subsequent intraperitoneal (i.p.) sensitization followed by intranasal challenge with OVA. A slow‐release pellet of 1‐MT implanted into mice was used to block IDO activity prior to repeated intranasal inoculation of OVA. We measured T‐cell proliferation in response to OVA, determined airway inflammation, and measured AHR to intranasal methacholine to investigate the role of IDO in sensitization to OVA.

Results:  Repeated intranasal administration of OVA generated tolerance and prevented a subsequent sensitization to OVA via the i.p. route. This response was inhibited in mice receiving a slow‐release pellet of 1‐MT. However, we successfully reconstituted tolerance in mice receiving 1‐MT following intra‐peritoneal injection of a mixture of kynurenine and hydroxyanthranilic acid.

Conclusion:  Our data suggest that, in addition to their role in IFN‐γ‐mediated inhibition of allergic airway inflammation, products of tryptophan catabolism play an important role in the prevention of sensitization to potential allergens in the respiratory airway.
Analysis of genotype by environment interaction for milk yield traits in first lactation of Simmental cattle
Authors: Gerber A. , Krogmeier D. , Emmerling R. , Götz K.‐U.
Published: August 2008

The breeding goal for Simmental cattle is derived for intensively managed dairy farms. Its suitability for extensive farms was addressed by analysing possible genotype by environment interaction (G × E) between the management levels for first lactation milk yield traits. A first analysis was performed with the data collected from 300 000 purebred daughters of 278 second crop bulls born in Bavaria in 1993 and 1994. The farms were classified by herd‐year‐effect, using the sum of fat and protein yields into two levels of management, either with 33 or 10% quantiles, corresponding to approximately 100 000 cows and 30 000 cows, respectively. The comparison was based on ‘daughter yield’ deviations (DYD). Correlations between DYD of extensive and intensive environments were 0.90, 0.91 and 0.87 for milk, fat and protein yield (kg) for 33% quantiles, respectively. Corresponding correlations for 10% quantiles were 0.85, 0.83 and 0.77. Despite high correlations, 50 out of 149 sires showed significant differences between DYD in different environments. Bulls with higher DYD for milk yield on intensive farms were superior in all environments. For the second analysis extensive and intensive farms in northern and southern Bavaria were chosen at random. Approximately 20 000 cows in each management class were used for the estimation of genetic parameters. In both regions phenotypic and additive‐genetic variances were higher in the intensively managed herds. Likewise heritabilities were higher for fat and protein yield, but not for milk where higher heritabilities were observed in 33% quantiles. Genetic correlations between extensive and intensive environments were 0.97 and above (33% quantiles). Ten per cent quantiles led to lower genetic correlations (0.90–0.95). Although no serious re‐ranking effects of sires were evident, the scale effect and the differences in genetic parameters should be taken into consideration in practical breeding.
Multiple trait genetic evaluation of ewe traits in Icelandic sheep
Authors: Árnason T. , Jónmundsson J.V.
Published: August 2008

The prolificacy of the ewes was measured as the number of lambs born per ewe mated (NLB) when the ewes were 1–4 years of age. The ewe productivity related to the same age interval was measured by special ewe production indices (EPI). The genetic parameters for these traits were estimated by a series of bivariate REML analyses using animal models. The material used for the genetic analysis contained records on 193 213 ewes. The heritability estimates for NLB were h2 = 0.17, 0.13, 0.11, 0.10 for the four respective age classes. Corresponding estimates for EPI were h2 = 0.16, 0.17, 0.17, 0.15. The genetic correlations among NLB at different ages ranged from 0.63 to 0.98 and among EPI from 0.82 to 0.99. The genetic correlations between NLB and EPI were generally low. The material used for estimating the breeding values by the MT‐BLUP Animal Model consisted of 1.5 million individuals in the pedigree file. In total 815 782 ewes had records for the NLB and 763 491 ewes had production index (at least 1 year). The records were registered in the years 1990–2006. All possible missing patterns were present in the data. In the iteration process expected values for missing traits were generated and solutions were obtained on canonical transformed scale. The genetic evaluations were run independently for NLB and EPI for computational convenience given the correlations between these traits were negligible.
Optimized management of genetic variability in selected pig populations
Authors: Colleau J.J. , Tribout T.
Published: August 2008

Controlling the increase of coancestry and inbreeding coefficients in selected populations is made possible through calculation of the optimal contributions allowed to breeding animals, given the current situation with regard to genetic diversity, and further, through optimal design of matings. The potential of such an approach for pig breeding was tested by retrospective optimization on the French Landrace population in reference to the matings actually carried out during a 21‐week test period. The major constraint was that the average overall estimated breeding value (EBV) should be the same as the observed one, for not decreasing short‐term genetic gain. Optimizing breeding allocations to boars would have led one to decrease coancestry and inbreeding coefficients by approximately 20%. This decrease would have even increased to approximately 30%, would have replacements and disposals been optimized after accounting for genetic variability, keeping the same constraint of genetic level identical to the observed one. These results showed the potential value, in the future, of completing each periodical calculation of EBVs by optimizations considering genetic variability and of releasing corresponding information to breeders, in order to enhance maintenance of genetic variability.
Transgenic zebrafish expressing fluorescent proteins in central nervous system neurons
Authors: Higashijima Shin‐ichi
Published: August 2008

Zebrafish is a powerful model system for investigations of vertebrate neural development. The animal has also become an important model for studies of neuronal function. Both in developmental and functional studies, transgenic zebrafish expressing fluorescent proteins in central nervous system neurons have been playing important roles. We review here the methods for producing transgenic zebrafish. Recent advances in transposon‐ or bacterial artificial chromosome‐based transgenesis greatly facilitate the creation of useful lines. We also present our study on alx‐positive neurons to reveal how transgenic zebrafish expressing fluorescent proteins in a specific class of neurons can be used to investigate their development and function.
In vivo DNA electrotransfer into muscle
Authors: Miyazaki Satsuki , Miyazaki Jun‐Ichi
Published: August 2008

Naked plasmid DNA injected into skeletal muscle is taken up by muscle cells and the genes in the plasmid are expressed. Among the non‐viral techniques for gene transfer in vivo, this method is especially simple, inexpensive, and safe. However, the relatively low expression levels attained by this method have limited its applications for uses other than as a DNA vaccine. We and other groups investigated the applicability of in vivo electroporation for gene transfer into muscle, using plasmid DNA vector. The results demonstrated that gene transfer into muscle by in vivo electroporation is far more efficient than simple intramuscular DNA injection and provides a potential approach to systemically delivering cytokines, growth factors, and other serum proteins for basic research and human gene therapy.
Gene transduction by sonoporation
Authors: Ohta Sho , Suzuki Kentaro , Ogino Yukiko , Miyagawa Shinichi , Murashima Aki , Matsumaru Daisuke , Yamada Gen
Published: August 2008

Gene transduction technologies are essential tools for understanding of gene functions or gene cascades underlying embryogenesis. In this review, we introduce a gene transduction method using microbubble and ultrasound (hereafter referred to as sonoporation). Sonoporation is carried out with relatively simple procedures and easily transduces genes into mesenchymal cells without significant damage to target tissues. Therefore, sonoporation is effective for gene transduction to study the molecular mechanisms of morphogenesis.
Practical guide of live imaging for developmental biologists
Authors: Kato Kagayaki , Hayashi Shigeo
Published: August 2008

Time‐lapse imaging of fluorescent proteins in living cells has become an indispensable tool in biological sciences. However, its application at the organismal level still faces a number of obstacles, such as large specimen sizes preventing illumination of internal tissues, high background fluorescence and uncontrollable movement of target tissues or embryos. Here we describe our solutions for these issues to obtain 4‐D fluorescent images from living Drosophila embryos using confocal microscopes. A computational procedure that detects and corrects the shift of moving objects to virtually stabilize them in time‐lapse movies (iSEMS) is presented. We discuss the importance of postimaging treatment of raw image stacks for the discovery of novel phenotypes that have previously escaped attention from the analyses of fixed specimens.
Electroporation as an efficient method of gene transfer
Authors: Odani Noritaka , Ito Kodai , Nakamura Harukazu
Published: August 2008

Gene transfer by electroporation is an indispensable method for the study of developmental biology, especially for the study using chick embryos. Here we briefly review the principles of the method, and its application to chick embryos. Methods of transient misexpression and long‐term misexpression by retrovirus vector or transposon system, and knockdown by small interference RNA are reviewed.
Application of heat shock promoter in transgenic zebrafish
Authors: Shoji Wataru , Sato‐Maeda Mika
Published: August 2008

The heat shock promoter is useful for regulating transgene expression in small water‐living organisms. In zebrafish embryos, downstream gene expression can be greatly induced throughout the body by raising the temperature from 28.5°C to 38.0°C. By manipulating the local temperature within an embryo, spatial control of transgene expression is also possible. One such way for inducing heat shock response in targeted cells is by using a laser microbeam under the microscope. In addition, random mosaic expression by transient gene expression and transplantation of the transgenic embryo into a wild type host can be considered a powerful tool for studying gene functions using this promoter. In this paper, we review the applications of the zebrafish heat shock protein promoter as a gene expression tool and for lineage labeling and transcription enhancer screening.
RNA interference by expressing short hairpin RNA in the Ciona intestinalis embryo
Authors: Nishiyama Aya , Fujiwara Shigeki
Published: August 2008

We carried out RNA interference by expressing short hairpin RNA (shRNA) in the Ciona intestinalis embryo. For this purpose, we identified a gene encoding U6 small nuclear RNA (snRNA) in the C. intestinalis genome. The 1‐kb sequence upstream of the U6 snRNA gene was sufficient for directing transcription of short RNA as revealed by Northern blot hybridization. An shRNA‐expressing plasmid vector was constructed, in which shRNA‐encoding oligonucleotides are inserted downstream of the U6 promoter. An shRNA that contained a sequence homologous to the C. intestinalis tyrosinase gene (Ci‐tyrosinase) suppressed melanization of pigment cells in the brain of morphologically normal tailbud embryos. An shRNA that perfectly matched the translated sequence of enhanced green fluorescent protein (EGFP) (a mutant type of Aequorea victoria green fluorescent protein) suppressed the expression of the coelectroporated EGFP transgene. These results suggest that the expression of shRNA interferes with functions of both endogenous and exogenous genes. The shRNA‐expressing plasmid constructed in the present study provides an easy and inexpensive alternative for the functional analysis of genes in ascidian embryos.
Gallera method of chick embryo culture in vitro supports better growth compared with original New method
Authors: Ghatpande Satish K.
Published: August 2008

An avian embryo is a valuable model system for vertebrate embryology. Easy availability, accessibility to various developmental stages and amenability of organ fields makes the chick embryo one of the favored model systems. Seminal discoveries regarding organogenesis and vertebrate morphogenesis have been made using chick embryos cultured in vitro. Dennis A.T. New revolutionized chick embryo culture methodology with his development of a single glass ring explantation technique. Many modifications and/or embellishments were introduced after the New era of embryo culture. A double glass ring method for chick embryo culture introduced by Gallera and Nicolet is compared with the original New method and the EASY method in this study. In addition, a video of culture methods is presented as a valuable tool in learning about and/or teaching techniques of chick embryo culture.
Do perineal exercises during pregnancy prevent the development of urinary incontinence? A systematic review
Authors: Lemos Andrea , de Souza Ariani Impieri , Ferreira Ana Laura Carneiro Gomes , Figueiroa José Natal , Cabral‐Filho José Eulálio
Published: August 2008

Objectives:  The aim of the current article was to conduct a systematic review of the performance of perineal exercises during pregnancy and their utility in the prevention of urinary incontinence.

Methods:  Randomized controlled studies (RCT) of a low‐risk obstetric population (primiparas or nulliparas) who had done perineal exercises only during pregnancy met the inclusion criteria. Articles published between 1966 and 2007 from periodicals indexed in the LILACS, SCIELO, PubMed/MEDLINE, SCIRUS and Cochrane Library databases were selected, using the following keywords: ‘urinary incontinence’, ‘pregnancy’, ‘pelvic floor’ and ‘exercise’. The Jadad scale was applied to assess the internal validity of the RCT and two meta‐analysis: one of fixed effects and the other of random effects were carried out with data extracted from the RCT, using the Stata 9.2 statistical software and adopting a significance level of 0.05.

Results:  Four RCTs with high methodological quality, involving a total of 675 women were included. They indicated that perineal muscle exercise significantly reduced the development of urinary incontinence from 6 weeks to 3 months after delivery (odds ratio = 0.45; confidence interval: 0.3 to 0.66). However, when evaluating this effect during the 34th and 35th gestational week, a meta‐analysis showed that the results were not significant (odds ratio = 0.13; confidence interval: 0.00 to 3.77).

Conclusion:  Pelvic floor muscle exercises may be effective at reducing the development of postpartum urinary incontinence, despite clinical heterogeneity among the RCT.
Effect of fasting during Ramadan on fetal development and maternal health
Authors: Dikensoy Ebru , Balat Ozcan , Cebesoy Bahar , Ozkur Ayhan , Cicek Hulya , Can Gunay
Published: August 2008

Aim:  The aim of the present study was to determine whether fasting during Ramadan causes ketonemia and/or ketonuria and their effects on fetal intrauterine development.

Methods:  Thirty‐six consecutive healthy women with uncomplicated pregnancies of ≥20 weeks of gestation who were fasting during Ramadan were included in the study group (group 1). The control group (group 2) consisted of 29 healthy pregnant women who were not fasting. Doppler ultrasonography was performed in all subjects in the beginning and at the end of Ramadan to evaluate the changes in the following measurements: fetal biparietal diameter; fetal femur length; and estimated fetal body weight. Fetal biophysical profile, amniotic fluid index, and umbilical artery systole/diastole ratio were measured in the beginning and at the end of Ramadan. Effects of fasting on the mother were evaluated by measuring serum concentrations of 3ß hydroxybutyrate and glucose, and urinary concentration of ketone. Subjects with any of the followings were excluded: diabetes; thyroid dysfunction; Cushing's syndrome; adrenal disease; pre‐eclampsia; and multiple pregnancy.

Results:  The mean duration of fasting in the study group was 18 ± 2.1 days. The mean maternal glucose level was significantly lower in the study group than in the control group (P = 0.003). No statistically significant differences were found between the two groups in the comparisons of other parameters.

Conclusion:  We concluded that fasting during Ramadan does not lead to maternal ketonemia or ketonuria in pregnant women. In addition, fasting during Ramadan has no significant adverse effect on intrauterine fetal development or the fetus's health.
Depth of invasion for appendiceal metastasis of ovarıan cancers: Does it have any clinical significance?
Authors: Ayhan Ali , Gultekin Murat , Sağlam Emine Arzu , Dursun Polat , Dogan Nasuh Utku , Aksan Guldeniz , Usubutun Alp , Yuce Kunter
Published: August 2008

Aim:  Depth of appendiceal metastasis in patients with epithelial ovarian carcinoma (EOC) has not been previously analyzed for a possible relationship with clinico‐pathological variables.

Methods:  Sixty‐nine patients with EOC treated at Hacettepe University were retrospectively analyzed. All of the patients had appendiceal metastasis. Pathological slides were re‐reviewed by the same pathologist. Appendiceal metastases were defined as serosal (if tumoral spread involved only serosa of the appendix) or sero‐mucosal (if tumoral spread also involved either muscular or mucosal surfaces of the appendix). Univariate and multivariate analysis did not reveal a significant difference with respect to the prognostic variables between the groups.

Results:  Thirty‐nine patients had serosal appendiceal metastasis (56.5%), while the remaining 30 patients (43.5%) had appendiceal metastasis extending toward the muscular layer (seromucosal metastasis: 16 within muscularis mucosa, 14 within the mucosa of the appendix). The mean age at the time of diagnosis was 54.58 years (range, 26‐88 years), with no significant difference between the groups (P = 0.9). Comparison of the survival rates between the two groups did not reveal a significant difference. Three‐year survival rates were 23.3% in the serosal metastasis and 27.9% in the seromucosal metastasis group (P = 0.9). This figure was 25% for patients with only muscular metastasis and 41.6% for patients with appendiceal metastasis extending to the mucosal layer (P = 0.2).

Conclusion:  This is the first report to analyze the metastatic pattern of EOC on the appendix with respect to depth of invasion which could not reveal a significant difference.
Serum ghrelin, leptin and resistin levels in adolescent girls with polycystic ovary syndrome
Authors: Bideci Aysun , Çamurdan Mahmut Orhun , Yeşilkaya Ediz , Demirel Fatma , Cinaz Peyami
Published: August 2008

Aim:  The aim of the present study was to investigate the levels of leptin, resistin and ghrelin in polycystic ovary syndrome (PCOS), and to assess their possible correlations with the hormonal and metabolic features of PCOS.

Methods:  Sixteen obese (ObPCOS) and 12 lean (LeanPCOS) subjects with PCOS and 19 obese control subjects were enrolled in the study.

Results:  Ghrelin, leptin and resistin concentrations were similar between groups when body mass index (BMI) was used as a covariate (P > 0.05). Mean androgen, SHBG, luteinizing hormone (LH) levels and luteinizing hormone/follicle‐stimulating hormone (LH/FSH) ratio tended to be similar between polycystic ovary syndrome (PCOS) groups. However, when compared with the control group, SHBG was lower and androgen, LH levels and LH/FSH ratio were higher in the PCOS groups. Free testosterone levels significantly correlated with resistin (r = −0.38), SHBG correlated significantly with body mass index (BMI) (r = −0.45) and resistin (r = −0.67), LH/FSH ratio was significantly correlated with ghrelin (r = −0.52) and estradiol (E2) levels (r = 0.51).

Conclusion:  ObPCOS and LeanPCOS groups having higher LH/FSH ratios and lower SHBG levels suggest that there could be factors other than adiposity responsible for the clinical features of PCOS patients. In the light of our results, those factors can be suggested as ghrelin and E2 for the elevated LH/FSH ratio and resistin for the lowered SHBG.
Unresolved issues in the management of ulcers of the foot in diabetes
Authors: Jeffcoate W. J. , Lipsky B. A. , Berendt A. R. , Cavanagh P. R. , Bus S. A. , Peters E. J. G. , Van Houtum W. H. , Valk G. D. , Bakker K. ,   on behalf of the three systematic review working parties of the International Working Group on the Diabetic Foot
Published: August 2008

Management of diabetic foot ulcers presents a major clinical challenge. The response to treatment is often poor and the outcome disappointing, while the costs are high for both healthcare providers and the patient. In such circumstances, it is essential that management should be based on firm evidence and follow consensus. In the case of the diabetic foot, however, clinical practice can vary widely. It is for these reasons that the International Working Group on the Diabetic Foot has published guidelines for adoption worldwide. The Group has now also completed a series of non‐systematic and systematic reviews on the subjects of soft tissue infection, osteomyelitis, offloading and other interventions designed to promote ulcer healing. The current article collates the results of this work in order to demonstrate the extent and quality of the evidence which is available in these areas. In general, the available scientific evidence is thin, leaving many issues unresolved. Although the complex nature of diabetic foot disease presents particular difficulties in the design of robust clinical trials, and the absence of published evidence to support the use of an intervention does not always mean that the intervention is ineffective, there is a clear need for more research in the area. Evidence from sound clinical studies is urgently needed to guide consensus and to underpin clinical practice. It is only in this way that patients suffering with these frequently neglected complications of diabetes can be offered the best hope for a favourable outcome, at the least cost.
Quality of life in children with Type 1 diabetes: a comparison of general and diabetes‐specific measures and support for a unitary diabetes quality‐of‐life construct
Authors: Nansel T. R. , Weisberg‐Benchell J. , Wysocki T. , Laffel L. , Anderson B. ,   for the Steering Committee of the Family Management of Diabetes Study
Published: August 2008

Aims  To assess the factor structure of the Pediatric Quality of Life Inventory (PedsQL) Diabetes Module and to compare the PedsQL general and diabetes‐specific quality of life (QOL) measures regarding psychometric properties and relations to relevant outcomes.

Methods  The instruments were completed by 447 children age 9 to 15.5 years with Type 1 diabetes > 1 year from four US paediatric diabetes clinics; parents completed the parallel parent‐proxy measures. Principal components factor analysis was used to examine the factor structure of the PedsQL diabetes module. Analyses of the generic and diabetes QOL measures included psychometric properties, parent–child correlations and correlations with depression, adherence and glycated haemoglobin (HbA1c).

Results  The factor structure of the PedsQL diabetes module did not support the original five subscales. Both one‐ and two‐factor models were supported; however, parallel parent and child subscales did not emerge. While the generic and diabetes‐specific measures of QOL were moderately to highly correlated with each other, the constructs were differentially associated with relevant diabetes outcomes. Generic QOL was more highly associated with depression than diabetes QOL. Conversely, diabetes QOL was more highly associated with adherence and HbA1c, although this was seen to a greater extent for parent‐proxy report than for child report.

Conclusions  Factor analysis of the PedsQL diabetes module supports the use of a total diabetes QOL score. Findings regarding the associations of the generic and diabetes modules with diabetes outcomes underscore the unique contribution provided by both generic and diabetes QOL.
Serum Th1 (CXCL10) and Th2 (CCL2) chemokine levels in children with newly diagnosed Type 1 diabetes: a longitudinal study
Authors: Antonelli A. , Fallahi P. , Ferrari S. M. , Pupilli C. , D’Annunzio G. , Lorini R. , Vanelli M. , Ferrannini E.
Published: August 2008

Aims   Cell‐mediated immunity and pro‐inflammatory cytokines are implicated in the pathogenesis of Type 1 diabetes. The aim of this study was to investigate whether circulating chemokines involved in T‐helper 1 (CXCL10) and T‐helper 2 (CCL2) autoimmunity are increased in children with Type 1 diabetes at onset and follow‐up.

Methods   Serum CXCL10 and CCL2 were measured in 96 children with newly diagnosed Type 1 diabetes, 59 age‐matched first‐degree relatives of diabetic children and 40 age‐matched non‐diabetic children with no family history of diabetes. In the diabetic children, an additional serum sample was obtained a median of 16 months after diagnosis.

Results   Serum CXCL10 levels were significantly higher in Type 1 children than in relatives or control children (P < 0.001); 44.7% of patients had a serum CXCL10 level ≥ 2 standard deviation above the mean value of the control group vs. 3.4% of relatives (P < 0.0001). In contrast, serum CCL2 levels were similar in patients, relatives and control subjects. In the Type 1 diabetic patients at follow‐up, CXCL10 was significantly reduced vs. baseline (P = 0.01), while CCL2 did not change.

Conclusions   In children with newly diagnosed Type 1 diabetes, raised serum CXCL10 and normal CCL2 concentrations signal a predominant T‐helper 1‐driven autoimmune process, which shifts toward T‐helper 2 immunity over the first 1–2 years from diagnosis.