Carrion flies of forensic interest: a study of seasonal community composition and succession in Lisbon,Portugal

Information on Diptera community, seasonality and successional patterns in every geographical region is fundamental for the use of flies as forensic indicators of time of death. In order to obtain these data from the Lisbon area (Portugal), experiments were conducted during the four seasons of the year, using piglet carcasses as animal models. Five stages were recognized during the decomposition process. The stages, besides visually defined, could be separated taking into account the occurrence and abundance of the specific groups of Diptera collected. In general, the bloated stage recorded higher abundance and species‐richness values. Seventy‐one species were identified, belonging to 39 families, in a total of 20 144 adult Diptera collected. Autumn yielded the highest values of species richness, whereas winter had the lowest. In all seasons of the year, Calliphoridae was the dominant family; Muscidae and Fanniidae were very abundant as well. Calliphora vicina Robineau‐Desvoidy, Calliphora vomitoria (Linnaeus), Chrysomya albiceps (Wiedemann), Lucilia ampullacea Villeneuve, Lucilia caesar (Linnaeus), Lucilia sericata (Meigen) (Calliphoridae), Hydrotaea ignava (Harris), Muscina prolapsa (Harris), Synthesiomyia nudiseta (van der Wulp) (Muscidae), Piophila megastigmata McAlpine, Stearibia nigriceps (Meigen) (Piophilidae) and Nemopoda nitidula (Fallén) (Sepsidae) were revealed to be very important members of the Diptera community collected. The necrophagous behaviour, demonstrated by their immatures, using carrion as a food source makes them useful forensic indicator species. Also of relevance is the presence of Chrysomya megacephala (Fabricius), S. nudiseta and P. megastigmata, foreign species established in the local carrion communities. This study also marks the first record of S. nudiseta in Portugal.     

High-resolution three-dimensional structure of horse heart cytochrome c

The 1.94 A resolution three-dimensional structure of oxidized horse heart cytochrome c has been elucidated and refined to a final R-factor of 0.17. This has allowed for a detailed assessment of the structural features of this protein, including the presence of secondary structure, hydrogen-bonding patterns and heme geometry. A comprehensive analysis of the structural differences between horse heart cytochrome c and those other eukaryotic cytochromes c for which high-resolution structures are available (yeast iso-1, tuna, rice) has also been completed. Significant conformational differences between these proteins occur in three regions and primarily involve residues 22 to 27, 41 to 43 and 56 to 57. The first of these variable regions is part of a surface beta-loop, whilst the latter two are located together adjacent to the heme group. This study also demonstrates that, in horse cytochrome c, the side-chain of Phe82 is positioned in a co-planar fashion next to the heme in a conformation comparable to that found in other cytochromes c. The positioning of this residue does not therefore appear to be oxidation-state-dependent. In total, five water molecules occupy conserved positions in the structures of horse heart, yeast iso-1, tuna and rice cytochromes c. Three of these are on the surface of the protein, serving to stabilize local polypeptide chain conformations. The remaining two are internally located. One of these mediates a charged interaction between the invariant residue Arg38 and a nearby heme propionate. The other is more centrally buried near the heme iron atom and is hydrogen bonded to the conserved residues Asn52, Tyr67 and Thr78. It is shown that this latter water molecule shifts in a consistent manner upon change in oxidation state if cytochrome c structures from various sources are compared. The conservation of this structural feature and its close proximity to the heme iron atom strongly implicate this internal water molecule as having a functional role in the mechanism of action of cytochrome c.     

Mediator structure and conformation change

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Ultrastructural Studies of Spermiogenesis of Conocephalus saltator (Saussure)

The acrosome of the spermatozoa is the organelle involved in its penetration through the ova membranes during the fertilization process. Several features of this process are considered to be related to the fertilization events e.g. some substance coats outside the membrane. During the maturation process the spermatozoa of Conocephalus saltator develop a coat of tubules and filaments which overly some membrane regions in a specific array. The mature spermatozoa are seen to adhere in these regions and form threads of 20 cells long and five to six wide. It is suggested that the external coat plays a role in this 'sticking' phenomenon.     

Responses of tropical cladocerans to a gradient of resource quality

1. The response of three tropical cladocerans to a gradient of resource quality was compared in a series of growth bioassays using seston collected from five lakes of different depth and trophic structure in Michigan, U.S.A. To assess the food quality in terms of digestibility, assimilation experiments were performed with ³²P‐labelled seston from the same lakes. Animals were also analysed for P‐content in their tissues at the end of these assays. 2. In general, assimilation efficiency was higher when animals fed on seston from shallow compared to deep lakes, and was significantly correlated with growth rates, suggesting that shallow lakes have the best food resources in terms of digestibility and P availability. 3. Results also showed that all cladoceran species responded similarly to the resource gradient, with lower growth rates in deep lakes and higher growth rates in shallow lakes, although the strength of response (sensitivity) was different among the species tested. 4. The cladoceran Moina micrura was the most sensitive species, and also displayed the highest P‐content and maximal growth rate, a pattern consistent with the growth rate hypothesis. 5. However, seston C : P ratio and growth rates in the different resources did not correlate with the animals’ P‐contents, showing an uncoupling between RNA‐phosphorus demands for growth and seston food quality. 6. In conclusion, our results support the idea that digestion resistance in algae is a major constraint to cladocerans in natural plankton communities.     

用RACE技术扩增并克隆牛BMP4基因3′端序列

RACE技术是一项扩增基因末端序列的新技术。该研究从牛BMP4基因出发,以牛软骨的RNA为模板,按照不同物种BMP4基因的相似性设计特异引物,运用PCR和RACE技术扩增并获得了特异片段,该片段经PCR、酶切和测序验证,证实所克隆序列为牛BMP4的3′端序列,包含有1170bp组成的开放读码框(ORF),编码389个氨基酸,3′非编码区121bp个核苷酸和poly(A)15。同源性分析结果表明,牛BMP4 cDNA最大开放读码框所推测的氨基酸序列与已知人、小鼠、大鼠、狗、羊和鸡等真核生物BMP4氨基酸序列进行比较,分别有94.5%、93.1%、91.9%、87.4%、94.2%、79%的同源性。这为克隆其他物种的BMP4基因提供了依据,同时牛骨形态发生蛋白的测序为我们更好的理解牛的生骨机理提供帮助。