The Interaction of Equine Lysozyme:Oleic Acid Complexes with Lipid Membranes Suggests a Cargo Off-Loading Mechanism

The normal function of equine lysozyme (EL) is the hydrolysis of peptidoglycan residues of bacterial cell walls. EL is closely related to α-lactalbumins with respect to sequence and structure and further possesses the calcium binding site of α-lactalbumins. Recently, EL multimeric complexes with oleic acids (ELOAs) were shown to possess tinctorial and morphological properties, similar to amyloidal aggregates, and to be cytotoxic. ELOA's interactions with phospholipid membranes appear to be central to its biological action, similar to human α-lactalbumin made lethal to tumor cells. Here, we describe the interaction of ELOA with phospholipid membranes. Confocal scanning laser microscopy shows that ELOA, but not native EL, accumulates on the surface of giant unilamellar vesicles, without inducing significant membrane permeability. Quartz crystal microbalance with dissipation data indicated an essentially non-disruptive binding of ELOA to supported lipid bilayers, leading to formation of highly dissipative and “soft” lipid membrane; at higher concentrations of ELOA, the lipid membrane desorbs from the surface probably as bilayer sheets of vesicles. This membrane rearrangement occurred to a similar extent when free oleic acid (OA) was added, but not when free OA was removed from ELOA by prior incubation with bovine serum albumin, emphasizing the role of OA in this process. NMR data indicated an equilibrium between free and bound OA, which shifts towards free OA as ELOA is progressively diluted, indicating that OA is relatively loosely bound. Activity measurements together with fluorescence spectroscopy and circular dichroism suggested a conversion of ELOA towards a more native-like state on interaction with lipid membranes, although complete refolding was not observed. Altogether, these results suggest that ELOA may act as an OA carrier and facilitate OA transfer to the membrane. ELOA's properties illustrate that protein folding variants may possess specific functional properties distinct from the native protein.     

Generation of mouse conditional and null alleles of the type III sodium‐dependent phosphate cotransporter PiT‐1

Accelerated vascular calcification occurs in several human diseases including diabetes and chronic kidney disease (CKD). In patients with CKD, vascular calcification is highly correlated with elevated serum phosphate levels. In vitro, elevated concentrations of phosphate induced vascular smooth muscle cell matrix mineralization, and the inorganic phosphate transporter‐1 (PiT‐1), was shown to be required. To determine the in vivo role of PiT‐1, mouse conditional and null alleles were generated. Here we show that the conditional allele, PiT‐1^flox, which has loxP sites flanking exons 3 and 4, is homozygous viable. Cre‐mediated recombination resulted in a null allele that is homozygous lethal. Examination of early embryonic development revealed that the PiT‐1^{Δe3,4/Δe3,4} embryos displayed anemia, a defect in yolk sac vasculature, and arrested growth. Thus, conditional and null PiT‐1 mouse alleles have been successfully generated and PiT‐1 has a necessary, nonredundant role in embryonic development. genesis 47:858–863, 2009. © 2009 Wiley‐Liss, Inc.     

Photoperiodic and Thermal Regulation of Development and Cold Hardiness in Larvae of the Clover Leaf Weevil, Hypera punctata

Effects of photoperiod and temperature on the development and cold hardiness were investigated in larvae of Hypera punctata. At a relatively low temperature (15°C), the larvae fed less and developed more slowly under a 12L:12D (SD) photoperiod than under a 16L:8D photoperiod (LD). SD larvae had lower gut weight against the whole body weight and lower supercooling point (SCP) than the LD counterparts for the same instar and same body weight. This was because the larval SCP is markedly affected by the quantity of the gut content. Laboratory experiments indicated that the low temperature mortality of this larvae occurred mainly due to freezing irrespective of the photoperiod and temperature, suggesting that the lower lethal temperature (LLT) depends on the supercooling ability of larvae. The SD larvae tended to have a lower SCP and hence a lower LLT than the LD counterparts at 15 or 10°C, unlike at 20°C. Thus, the slower larval development under SD conditions at relatively low temperatures may prevent larvae from reaching the later instar, which have a higher SCP and thus less cold tolerance, during the coldest season. The suppressed feeding activity under SD conditions would lower the SCP, thereby reducing the possibility of lethal tissue freezing. Such a photoperiodic and thermal regulation of the larval development and the supercooling ability appear to represent adaptive mechanisms for winter survival in this beetle.     

Population persistence under high mutation rate: From evolutionary rescue to lethal mutagenesis

Populations may genetically adapt to severe stress that would otherwise cause their extirpation. Recent theoretical work, combining stochastic demography with Fisher's geometric model of adaptation, has shown how evolutionary rescue becomes unlikely beyond some critical intensity of stress. Increasing mutation rates may however allow adaptation to more intense stress, raising concerns about the effectiveness of treatments against pathogens. This previous work assumes that populations are rescued by the rise of a single resistance mutation. However, even in asexual organisms, rescue can also stem from the accumulation of multiple mutations in a single genome. Here, we extend previous work to study the rescue process in an asexual population where the mutation rate is sufficiently high so that such events may be common. We predict both the ultimate extinction probability of the population and the distribution of extinction times. We compare the accuracy of different approximations covering a large range of mutation rates. Moderate increase in mutation rates favors evolutionary rescue. However, larger increase leads to extinction by the accumulation of a large mutation load, a process called lethal mutagenesis. We discuss how these results could help design “evolution‐proof” antipathogen treatments that even highly mutable strains could not overcome.     

Changements de l'epithélium médian des bourgeons palatins de souris au stade de préfusion

Résumé Avant d'entrer en contact, les surfaces de fusion des processus palatins subissent des modifications. Certaines cellules épithéliales superficielles deviennent turgescentes, alors que d'autres conservent un aspect dense et aplati, elles se lysent et éclatent. Des noyaux semblables à ceux des assises profondes se retrouvent en surface, paraissant migrer vers le bourgeon antagoniste. Des projections filamenteuses apparaissent, quelques unes établissant un contact avec l'épithélium opposé, d'autres restant seulement sous forme d'expansion. Ces observations sont concordantes en microscopie photonique et en microscopie électronique à balayage.

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Modification of the medial epithelium of the palatal shelves of mice at the prefusion stage. A light and scanning electron microscopy study

Summary The fusion surfaces of the palatal processes are modified before coming into contact. Several superficial epithelial cells become hypertrophied, undergo lysis and burst, whereas others remain flattened and densely stained. Nuclei, similar to those localized in deeper layers are found on the surface and seem to migrate towards the opposite process. Filamentous projections appear and some establish a linkage with the opposite epithelium, whereas others expand freely from the surface. Light microscope observations agree with scanning electron microscopy findings.

     

绿原酸提高舞毒蛾核型多角体病毒(LdNPV)的致病力

【目的】明确绿原酸(chlorogenic acid,CA)对舞毒蛾核型多角体病毒Lymantria dispar necleopolyhydrovirus(Ld NPV)致病力的影响,为舞毒蛾的防治提供参考依据。【方法】采用食料给毒法进行生物测定,测定舞毒蛾2龄幼虫对单独Ld NPV及添加绿原酸的Ld NPV(CA+Ld NPV)的剂量及致死时间响应。【结果】CA+Ld NPV与单独Ld NPV对舞毒蛾2龄幼虫的剂量及时间响应间有显著差异,二者对舞毒蛾2龄幼虫的致死中浓度(LC50)分别为161.8 OBs/μL(95%的置信区间为105.6~235.3 OBs/μL)和264.4 OBs/μL(95%的置信区间为178.6~384.0 OBs/μL),前者对舞毒蛾的致病力较后者强。CA+Ld NPV对舞毒蛾2龄幼虫的致死中时间(LT_(50))较Ld NPV的短,当Ld NPV浓度为590 OBs/μL,CA+Ld NPV及Ld NPV对舞毒蛾2龄幼虫的LT_(50)分别为9.9 d和12.3 d;当Ld NPV浓度为5 900 OBs/μL时,则LT_(50)分别为6.9 d和8.0 d。绿原酸降低了Ld NPV对舞毒蛾幼虫的致死中浓度,缩短了致死中时间。【结论】绿原酸可提高Ld NPV对舞毒蛾2龄幼虫的致病力,其机理有待进一步研究。     

Mediterranean flour moth Ephestia kuehniella eggs and larvae exposed to a static magnetic field and preference by Trichogramma embryophagum

This study investigated the effect of strong magnetic fields as insecticidal activity on Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae) larvae and eggs at different stages of development and their preference by the egg parasitoid, Trichogramma embryophagum Hartig (Hymenoptera: Trichogrammatidae). Eggs ranging in age from 24-h to 48-h and 72-h-old and larvae (1 to 2 days) were exposed to 1.4 Tesla (T) magnetic fields from a DC power supply at 50 Hz for different time periods (3, 6, 12, 24, 48 and 72 h). Twelve hours of exposure at 1.4 T was toxic to 24-h-old eggs of E. kuehniella. The 72-h-old host eggs treated with 1.4 T for 6–72 h were not significantly preferred by T. embryophagum. The magnetic field was toxic to 24-h-old eggs of E. kuehniella exposed to 1.4 T for 12. The treatment of magnetic fields on the 72-h-old host egg with 1.4 T at 6–72 h was not significantly preferred by T. embryophagum. Magnetization of 24-h-old eggs of E. kuehniella for 3 h could be effectively used with T. embryophagum as sterilised host eggs. These eggs were markedly preferred by T. embryophagum. The LT₅₀ and LT₉₉ values of magnetic fields at different egg stages of E. kuehniella, and larvae were measured. A level of 1.4 T at 72 h completely prevented the development of the larvae. There was no significant effect on larval survival at 1.4 T at 48 and 72 h. Increasing magnetic fields exposure times for eggs that were 24-h, 48-h and 72-h-old prevented larval emergence and increased their mortality rate. Consequently, magnetic fields could be used in controlling stored-product pest eggs and larvae of E. kuehniella.     

Air and surface soil samples – two different pairs of shoes? Comparing the pollen spectrum on different days of the pollen season

The pollen spectra of air and surface soil samples from a rooftop (at 14 m) and from ground level (at 1.6 m) in the suburbs of Vienna (Austria) were compared. Two soil samples and two air samples were taken on four different days to account for possible differences: in winter when no pollination occurred (reference day), in spring during the main flowering of Betula (birch day), in spring/summer during the main flowering of Poaceae (grass day), and in autumn during the main flowering of Ambrosia (ragweed day). Thirty-five different pollen types were used to describe the pollen spectra. Frequencies of certain pollen types reflect a seasonal impact on both the surface soil and air samples and show a similarity between air and soil samples on most of the days. However, the seasonal impact is higher in the air samples and shows a high consistency for ground and rooftop level. Kendall’s tau correlation coefficients further substantiate the similarities of the samples especially for the pollen season days. Exceptions include the winter day when pollination was low and the air samples recorded nearly no pollen at all, and the ragweed day when Ambrosia pollen was abundant in three of four samples but not in the ground surface soil sample. Thus, (1) air and surface pollen samples record similar signals during the pollen season but not during the ragweed and winter season and (2) air and surface pollen samples show the impact of local vegetation also in pollen traps located at different heights.