Insect morphology in the age of phylogenomics: innovative techniques and its future role in systematics

A brief account of the history of insect morphology is given. Different techniques and analytical methods used in current projects on insect morphology and phylogeny and their optimized combined application are described. These include fixation, dissection, maceration, histology (microtome sectioning), scanning electron microscopy (SEM), transmission electron microscopy (TEM), serial block‐face scanning electron microscopy (SBFSEM), focused ion beam scanning electron microscopy (FIB/SEM), confocal laser scanning microscopy (CLSM), bleaching, micro‐computed tomography (μCT), computer‐based three‐dimensional reconstruction, focus stacking of digital images, geometric morphometrics and the storage of morphological metadata. The role of insect morphology in the “age of phylogenomics” is discussed.     

Micro‐organisms in latex and natural rubber coagula of Hevea brasiliensis and their impact on rubber composition,structure and properties

Natural rubber, produced by coagulation of the latex from the tree Hevea brasiliensis, is an important biopolymer used in many applications for its outstanding properties. Besides polyisoprene, latex is rich in many nonisoprene components such as carbohydrates, proteins and lipids and thereby constitutes a favourable medium for the development of micro‐organisms. The fresh rubber coagula obtained by latex coagulation are not immediately processed, allowing the development of various microbial communities. The time period between tree tapping and coagula processing is called maturation, during which an evolution of the properties of the corresponding dry natural rubber occurs. This evolution is partly related to the activity of micro‐organisms and to the modification of the biochemical composition. This review synthesizes the current knowledge on microbial populations in latex and natural rubber coagula of H. brasiliensis and the changes they induce on the biochemistry and technical properties of natural rubber during maturation.     

Microdamage assessment of bone-cement interfaces under monotonic and cyclic compression

Bone-cement interface has been investigated under selected loading conditions, utilising experimental techniques such as in situ mechanical testing and digital image correlation (DIC). However, the role of bone type in the overall load transfer and mechanical behaviour of the bone-cement construct is yet to be fully quantified. Moreover, microdamage accumulation at the interface and in the cement mantle has only been assessed on the exterior surfaces of the samples, where no volumetric information could be obtained. In this study, some typical bone-cement interfaces, representative of different fixation scenarios for both hip and knee replacements, were constructed using mainly trabecular bone, a mixture of trabecular and cortical bone and mainly cortical bone, and tested under static and cyclic compression. Axial displacement and strain fields were obtained by means of digital volume correlation (DVC) and microdamage due to static compression was assessed using DVC and finite element (FE) analysis, where yielded volumes and strains (ε_zz) were evaluated. A significantly higher load was transferred into the cement region when mainly cortical bone was used to interdigitate with the cement, compared with the other two cases. In the former, progressive damage accumulation under cyclic loading was observed within both the bone-cement interdigitated and the cement regions, as evidenced by the initiation of microcracks associated with high residual strains (ε_{zz_res}).     

HS‐SPME‐GC‐FID method for detection and quantification of Bacillus cereus ATCC 10702 mediated 2‐acetyl‐1‐pyrroline

A rapid micro‐scale solid‐phase micro‐extraction (SPME) procedure coupled with gas‐chromatography with flame ionized detector (GC‐FID) was used to extract parts per billion levels of a principle basmati aroma compound “2‐acetyl‐1‐pyrroline” (2‐AP) from bacterial samples. In present investigation, optimization parameters of bacterial incubation period, sample weight, pre‐incubation time, adsorption time, and temperature, precursors and their concentrations has been studied. In the optimized conditions, detection of 2‐AP produced by Bacillus cereus ATCC10702 using only 0.5 g of sample volume was 85 μg/kg. Along with 2‐AP, 15 other compounds produced by B. cereus were also reported out of which 14 were reported for the first time consisting mainly of (E)?2‐hexenal, pentadecanal, 4‐hydroxy‐2‐butanone, n‐hexanal, 2–6‐nonadienal, 3‐methoxy‐2(5H) furanone and 2‐acetyl‐1‐pyridine and octanal. High recovery of 2‐AP (87 %) from very less amount of B. cereus samples was observed. The method is reproducible fast and can be used for detection of 2‐AP production by B. cereus. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1356–1363, 2014     

菹草茎叶微界面O2时空动态

采用高分辨率光纤氧微电极测定了富营养化水体中沉水植物菹草(Potamogeton crispus)茎叶微界面(0—2.0 mm)氧(O_2)。菹草叶微界面O_2浓度梯度具明显的时空变化。时间上,菹草叶微界面O_2浓度具有明显的生长阶段变化和昼夜变化。幼苗期和快速生长期微界面O_2浓度增加幅度较小,稳定期叶表O_2浓度梯度增加幅度最大,衰亡期叶微界面O_2浓度受附着物影响具明显的空间梯度。菹草叶微界面O_2表现为昼高夜低的单峰变化模式,主要受光照和温度的影响。空间上,越接近茎叶表面,O_2浓度越高。顶部幼叶微界面O_2浓度梯度增加较平缓,中部成熟叶微界面O_2浓度梯度变化最陡,波动幅度最大,中部茎和基部衰老叶微界面O_2浓度梯度由于受密集附着物的影响,在附着物表面达到最大值,进入附着层后略有下降。结果表明,菹草茎叶微界面O_2时空变化主要受附着物和植物光合放氧能力的影响。光纤微电极是一种分析植物叶微界面氧时空分布的理想工具,对深入研究植物微界面在富营养化水体中养分的迁移转化具有重要意义,可为水生植物生理生态研究提供有力工具。     

A biophotonic approach to measure pH in small volumes in vitro: Quantifiable differences in metabolic flux around the cumulus‐oocyte‐complex (COC)

Unfertilised eggs (oocytes) release chemical biomarkers into the medium surrounding them. This provides an opportunity to monitor cell health and development during assisted reproductive processes if detected in a non‐invasive manner. Here we report the measurement of pH using an optical fibre probe, OFP1, in 5 μL drops of culture medium containing single mouse cumulus oocyte complexes (COCs). This allowed for the detection of statistically significant differences in pH between COCs in culture medium with no additives and those incubated with either a chemical (cobalt chloride) or hormonal treatment (follicle stimulating hormone); both of which serve to induce the release of lactic acid into the medium immediately surrounding the COC. Importantly, OFP1 was shown to be cell‐safe with no inherent cell toxicity or light‐induced phototoxicity indicated by negative DNA damage staining. Pre‐measurement photobleaching of the probe reduced fluorescence signal variability, providing improved measurement precision (0.01‐0.05 pH units) compared to previous studies. This optical technology presents a promising platform for the measurement of pH and the detection of other extracellular biomarkers to assess cell health during assisted reproduction.     

3D heart morphological changes in response to developmental temperature in zebrafish: More than ventricle roundness

A new, three‐dimensional geometric morphometric approach was assessed to study the effect of developmental temperature on fish heart shape utilizing geometric morphometrics of three‐dimensional landmarks captured on digitally reconstructed zebrafish hearts. This study reports the first three‐dimensional analysis of the fish heart and demonstrates significant shape modifications occurring after three developmental temperature treatments (T_D = 24, 28 or 32°C) at two distinct developmental stages (juvenile and adult fish). Elevation of T_D induced ventricle roundness in juveniles, males and females. Furthermore, significant differences that have not been described so far in heart morphometric indices (i.e., ventricle sphericity, bulbus arteriosus elongation and relative location, heart asymmetry) were identified. Sex proved to be a significant regulating factor of heart shape plasticity in response to T_D. This methodology offers unique benefits by providing a more precise representation of heart shape changes in response to developmental temperature that are otherwise not discernable with the previously described two‐dimensional methods. Our work provides the first evidence of three‐dimensional shape alterations of the zebrafish heart adding to the emerging rationale of temperature‐driven plastic responses of global warming and seasonal temperature disturbances in wild fish populations and in other ectothermic vertebrates as well (amphibians and reptiles).     

Knockdown of AMPKα decreases ATM expression and increases radiosensitivity under hypoxia and nutrient starvation in an SV40-transformed human fibroblast cell line,LM217

Background

Presence of unperfused regions containing cells under hypoxia and nutrient starvation contributes to radioresistance in solid human tumors. It is well known that hypoxia causes cellular radioresistance, but little is known about the effects of nutrient starvation on radiosensitivity. We have reported that nutrient starvation induced decrease of mTORC1 activity and decrease of radiosensitivity in an SV40-transformed human fibroblast cell line, LM217, and that nutrient starvation induced increase of mTORC1 activity and increase of radiosensitivity in human liver cancer cell lines, HepG2 and HuH6 (Murata et al., BBRC 2015). Knockdown of mTOR using small interfering RNA (siRNA) for mTOR suppressed radiosensitivity under nutrient starvation alone in HepG2 cells, which suggests that mTORC1 pathway regulates radiosensitivity under nutrient starvation alone. In the present study, effects of hypoxia and nutrient starvation on radiosensitivity were investigated using the same cell lines.