Generation and characterization of Clostridium septicum alpha toxin mutants and their use in diagnosing paroxysmal nocturnal hemoglobinuria

Glycosylphosphatidylinositol (GPI) anchors various proteins to the membrane of eukaryotic cells. Paroxysmal nocturnal hemoglobinuria (PNH) is a hematopoietic stem cell disorder that is primarily due to the lack of GPI-anchored proteins on the surface of blood cells. To detect the GPI-deficient cells in PNH patients, we modified alpha toxin, a pore-forming toxin of the Gram-positive bacterium Clostridium septicum. We first showed that aerolysin, a homologous toxin from Aeromonas hydrophila, bound to both of Chinese hamster ovary cells deficient of N-glycan maturation as well as GPI biosynthesis at a significant level. However, alpha toxin bound to the mutant cells of N-glycosylation, but not to GPI-deficient cells. It suggested that alpha toxin could be used as a specific probe to differentiate only GPI-deficient cells. As a diagnostic probe, alpha toxin must be the least cytotoxic while maintaining its affinity for GPI. Thus, we constructed several mutants. Of these, the mutants carrying the Y155G or S189C/S238C substitutions bound to GPI as well as the wild-type toxin. These mutants also efficiently underwent proteolytic activation and aggregated into oligomers on the cell surface, which are events that precede the formation of a pore in the host cell membrane, leading to cell death. Nevertheless, these mutants almost completely failed to kill host cells. It was revealed that the substitutions affect the events that follow oligomerization. The S189C/S238C mutant toxin differentiated GPI-deficient granulocyte and PMN, but not red blood cells, of a PNH patient from GPI-positive cells at least as sensitively as the commercial monoclonal antibodies that recognize the CD59 or CD55 GPI proteins on blood cells. Thus, this modified bacterial toxin can be employed instead of costly monoclonal antibodies to diagnose PNH patients.     

华北落叶松夜间树干液流特征及生长季补水格局

在宁夏六盘山北侧半干旱区的叠叠沟小流域,采用热扩散探针法在2011年生长季监测了华北落叶松(Larix principisrupprechtii)人工林的树干液流速率,分析了夜间树干液流和补水量的变化特征及与气象、土壤水分等环境因子的关系.结果表明:树干液流速率日变化表现为典型的单峰宽峰曲线,且整个生长季均存在微弱的夜间液流,一般表现为逐渐减小,特别是在晴天,且晴天的变幅显著大于雨天.除生长季中期雨天夜间液流平均速率显著高于晴天,生长季初期及末期雨天与晴天的差异并不显著.生长季内,夜间树干补水总量为11.03 mm,占总蒸腾量的7.22％;5月份的树干补水量最大(4.19mm),其他月份的树干补水量明显减小,在0.9-1.7mm的范围波动.但不同月份间的补水贡献率存在明显差异,表现为生长季末期(9、10月)＞初期(5月)＞中期(6-8月).相关分析表明,日补水量与各气象因子关系不大,仅与降水量显著正相关(P＜0.05),与土壤含水率、日间蒸腾量、日蒸腾总量极显著正相关(P＜0.01).夜间补水的月蒸腾贡献率与月均土壤含水率、月均气温、月均日间蒸腾量、月总蒸腾量等显著相关(P＜0.05);而夜间补水的日蒸腾贡献率与日最高气温、日均气温、日间蒸腾量、日均饱和水汽压差、日总蒸腾量、日均太阳辐射强度、日最低气温、日均空气相对湿度、日降水量、土壤含水率等极显著相关(P＜0.01),经逐步回归分析建立了日补水量蒸腾贡献率与环境因子的多元线性模型.     

Wait Until Dark? Daily Activity Patterns and Nest Predation by Snakes

Predation involves costs and benefits, so predators should employ tactics that reduce their risk of injury or death and that increase their success at capturing prey. One potential way that predators could decrease risk and increase benefits is by attacking prey at night when risks may be reduced and prey more vulnerable. Because some snakes are facultatively nocturnal and prey on bird nests during the day and night, they are ideal for assessing the costs and benefits of diurnal vs. nocturnal predation. We used automated radiotelemetry and cameras to investigate predation on nesting birds by two species of snakes, one diurnal and the other facultatively nocturnal. We predicted that snakes preying on nests at night should experience less parental nest defence and capture more adults and nestlings. Rat snakes (Pantherophis obsoletus) were relatively inactive at night (23–36% activity) but nearly always preyed on nests after dark (80% of nest predations). Conversely, racers (Coluber constrictor) were exclusively diurnal and preyed on nests during the times of day they were most active. These results are consistent with rat snakes strategically using their capacity for facultative nocturnal activity to prey on nests at night. The likely benefit is reduced nest defence because birds defended their nests less vigourously at night. Consistent with nocturnal predation being safer, rat snake predation events lasted three times longer at night than during the day (26 vs. 8 min). Nocturnal nest predation did not make nests more profitable by increasing the likelihood of capturing adults or removing premature fledging of nestlings. The disconnect between rat snake activity and timing of nest predation seems most consistent with rat snakes locating prey during the day using visual cues but waiting until dark to prey on nests when predation is safer, although designing a direct test of this hypothesis will be challenging.     

Behavioral thermoregulation and the role of melatonin in a nocturnal snake

Daily and seasonal variations in hormone levels influence the complex interactions between behavior and physiology. Ectothermic animals possess the unique ability behaviorally to adjust body temperature (T(b)) to control physiological rate processes. Thus, a hormone may indirectly influence a physiological rate by directly influencing the behaviors that adjust or control that rate process. Although many hormonal influences on behavioral regulation of T(b) remain uninvestigated, melatonin (MEL) generally is considered a hormone that decreases mean preferred T(b). Many ectotherms demonstrate the selection of lower T(b)'s in response to increased MEL concentrations. Here, we examined the influence of MEL on the behavioral regulation of T(b) in the nocturnal African house snake Lamprophis fuliginosus. A series of experiments with two injection regimes of MEL had no significant effect on the mean preferred T(b) of L. fuliginosus. In addition, mean preferred T(b)'s during the photophase did not differ significantly from those during scotophase. Our findings suggest that L. fuliginosus does not respond to elevated concentrations of either endogenous or exogenous MEL. To verify that the African house snake is nocturnal, we investigated activity patterns of L. fuliginosus throughout the photoperiod. The activity period of L. fuliginosus occurs in the scotophase of the photoperiod, a pattern consistent with that of nocturnal species. This suggests that nocturnal organisms such as L. fuliginosus may not respond to MEL in the same manner as many diurnal species. Our results support the hypothesis that some animals, particularly nocturnal species, may have developed alternative responses to increased plasma concentrations of MEL.     

Decoding the ‘zeep’ complex: quantitative analysis of interspecific variation in the nocturnal flight calls of nine wood warbler species (Parulidae spp.)

ABSTRACT

The “zeep” complex consists of nine birds that produce nocturnal flight calls with similar acoustic features. Our inability to distinguish these calls inhibits the acoustic monitoring of these species. We test the hypothesis that flight calls of nine warblers in the “zeep” complex show sufficient acoustic differences to allow differentiation. We investigate divergence in these vocalizations by recording birds held for banding and collecting additional recordings from sound libraries. We used three approaches to compare calls between species: analysis of variance in acoustic properties, discriminant analysis of acoustic properties, and spectrographic cross-correlation. The first approach revealed five species that were different in one or more acoustic properties. The second approach revealed a level of assignment to the correct species (73%) that exceeded levels expected by chance (36%). The third approach revealed calls of seven species to be significantly more similar to conspecific calls than heterospecific calls. Our results suggest the calls of many members of the “zeep” complex exhibit species-specific differences in structure, which may allow differentiation of at least five “zeep” species based on call alone. We advocate for the combined use of these three approaches for the comparison of “zeep” calls in future flight call studies.     

Periodicity of infectivity of plasmodial gametocytes: the "Hawking phenomenon"

Five controlled experiments were performed to test the validity of the theory of nocturnal infectivity of gametocytes of Plasmodium. Two gave equivocal results probably because synchronicity of parasitaemia was low; the remaining three, in which a langur strain of P. cynomolgi was used, largely confirmed Hawking's work and in particular demonstrated the much greater production of oocysts after midnight as compared with midday feedings.     

Effects of Endurance Exercise on Nocturnal Hormone Concentrations in Males

A preliminary study was conducted to evaluate the change in nocturnal concentrations of testosterone, luteinizing hormone, growth hormone, prolactin, thyroxine, and Cortisol following a control day (no exercise) and on a day in which exercise was performed. Exercise consisted of 90 min of cycling at 70% of each subject's maximal oxygen uptake. The exercise occurred from 1630 to 1800 hr on the exercise day while a comparable period of rest took place on the control day. Hormonal concentrations were evaluated at 2-hr intervals for a 12-hr period each night (2000-0800 hr). The subjects slept from 2255 (± 20 min [x ± S.E.M.]) until 0715 (± 15) during each night. All hormone responses were plotted and integrated for the 12-hr period. Analysis indicated that a significant augmentation of the prolactin and thyroxine responses occurred, while concurrently an attenuation of the growth hormone and Cortisol responses were observed. Contrastingly, no significant effects were found for the testosterone and luteinizing hormones responses. The physiological significance of these findings remained to be determined, but the results do suggest that further research is warranted in the area.     

Understanding the function of nocturnal song in ovenbirds: males do not countersing at night

Many diurnal bird species vocalize at night, however the function of nocturnal song is, generally, still poorly understood. Previous research has suggested that nocturnal song may serve a social function and is influenced by environmental factors. To test whether males attend to the nocturnal song of conspecifics, we experimentally exposed ovenbirds Seiurus aurocapilla to nocturnal flight songs, and recorded their response both during the night and during the following dawn chorus. We compared latency to song and vocal output before and after playback exposure to determine if males altered their vocalizations in response to exposure to flight songs from an unknown male. We found no evidence of counter singing or change in nocturnal song output, nor a change in vocal output during the dawn chorus following playback exposure. Our results suggest that, in ovenbirds, nocturnal song does not serve as an intraspecific social function. Nocturnal song, through rare, may be significant in the mating systems of some diurnal bird species, and requires additional study.     

Use of the Calmset 3 Biofeedback/Relaxation System in the Assessment and Treatment of Chronic Nocturnal Bruxism

Biofeedback and nocturnal alarms using electromyographic (EMG) activity of the masseter muscles have often been used to treat nocturnal bruxism. However, although use of EMG activated nocturnal alarms has been successful, the clinical utility of the devices reported in the literature to date is questionable. For instance, many of the devices are cumbersome and specifically designed and constructed by the investigators. Hence, the purpose of the present investigation was to test the clinical utility of the Calmset (Thought Technology Limited, Montreal, Canada), a commercially available, user-friendly, compact, and portable EMG biofeedback instrument that may be used as an EMG activated nocturnal alarm. To meet this objective, the Calmset was used both to facilitate assessment and to treat an individual with chronic nocturnal bruxism. The results indicated that the patient exhibited fewer bruxing episodes following treatment and that treatment gains were maintained 6 months following termination of treatment. Advantages and disadvantages of using the Calmset are discussed.     

川西亚高山草甸土壤呼吸的昼夜变化及其季节动态

采用LI-8100A土壤碳通量自动测量系统,于2011年4—11月对川西亚高山草甸的土壤呼吸进行了测定,分析了水热因子对草甸昼、夜间土壤呼吸特征及其季节动态变化的影响。结果表明:1)川西亚高山草甸昼、夜间土壤呼吸的变化格局不同,昼间呈双峰型,夜间呈抛物线型;整个观测期(4—11月)内,夜间土壤呼吸占总土壤呼吸的46.79%,其中草盛期(6—9月)的昼、夜土壤呼吸占有较大比例。2)8次夜间土壤呼吸测定的变化趋势基本相同,最低值均在06:30—07:00左右;昼、夜间土壤呼吸季节变化的主要影响因素为土壤5 cm温度,Van’t Hoff指数模型和Lloyd和Taylor方程均适合预测整个的土壤呼吸通量。3)返青期(4—5月)和枯黄期(10—11月)的昼、夜间土壤呼吸差异不显著,均低于草盛期;整个观测期,草甸昼、夜间土壤呼吸与土壤温度均有显著的指数相关,而与土壤水分含量存在显著线性相关则表现在草返青期夜间及枯黄期昼、夜间。4)整个观测期昼、夜间的Q10值分别为3.90和3.74;对Q10值的分析表明,返青期昼土壤呼吸的Q10值最大(4.14),草盛期夜间的Q10值最小(1.40)。研究结果说明,采用昼、夜间土壤呼吸的瞬时值来推算土壤呼吸的季节通量和年通量时,不仅需要加大观测期间夜间土壤呼吸的测定,还需要考虑昼、夜间土壤温度和土壤含水量及其它生物因子的影响。     

Nocturnal incubation recess and flushing behavior by duck hens

Incubating birds must balance the needs of their developing embryos with their own physiological needs, and many birds accomplish this by taking periodic breaks from incubation. Mallard (Anas platyrhynchos) and gadwall (Mareca strepera) hens typically take incubation recesses in the early morning and late afternoon, but recesses can also take place at night. We examined nocturnal incubation recess behavior for mallard and gadwall hens nesting in Suisun Marsh, California, USA, using iButton temperature dataloggers and continuous video monitoring at nests. Fourteen percent of all detected incubation recesses (N = 13,708) were nocturnal and took place on 20% of nest‐days (N = 8,668). Video monitoring showed that hens covered their eggs with down feathers when they initiated a nocturnal recess themselves as they would a diurnal recess, but they left the eggs uncovered in 94% of the nocturnal recesses in which predators appeared at nests. Thus, determining whether or not eggs were left uncovered during a recess can provide strong indication whether the recess was initiated by the hen (eggs covered) or a predator (eggs uncovered). Because nest temperature decreased more rapidly when eggs were left uncovered versus covered, we were able to characterize eggs during nocturnal incubation recesses as covered or uncovered using nest temperature data. Overall, we predicted that 75% of nocturnal recesses were hen‐initiated recesses (eggs covered) whereas 25% of nocturnal recesses were predator‐initiated recesses (eggs uncovered). Of the predator‐initiated nocturnal recesses, 56% were accompanied by evidence of depredation at the nest during the subsequent nest monitoring visit. Hen‐initiated nocturnal recesses began later in the night (closer to morning) and were shorter than predator‐initiated nocturnal recesses. Our results indicate that nocturnal incubation recesses occur regularly (14% of all recesses) and, similar to diurnal recesses, most nocturnal recesses (75%) are initiated by the hen rather than an approaching predator.     

Diel activity patterns of juvenile Atlantic salmon in rivers with summer water temperature near the temperature‐dependent suppression of diurnal activity

Direct day and night underwater observations of juvenile Atlantic salmon Salmo salar during summer and autumn showed a duality in response to temperature between 7 and 11° C for young‐of‐the‐year (YOY) Atlantic salmon. They were predominantly diurnal in early summer and nocturnal in late summer although water temperatures were similar. Post‐YOY Atlantic salmon did not show a strong response to temperature as they were mostly nocturnal during the study period. It is suggested that the difference in activity patterns between YOY and post‐YOY Atlantic salmon can be explained by size‐dependent trade‐off between growth and predation risk.     

Diurnal and nocturnal ranging behaviour of Stone‐curlews Burhinus oedicnemus nesting in river habitat

Studying a species under a range of conditions is essential for fully understanding its ecology and for predicting its response to human impacts on the environment. We investigated the spatial behaviour and the habitat characteristics of foraging areas of Eurasian Stone‐curlews breeding in an important but poorly investigated habitat, gravel riverbed, throughout the full 24‐h cycle. The data collected for 17 radiotagged birds nesting in the Taro River Regional Park (Parma, Italy) showed a clear split between diurnal and nocturnal spatial behaviour. Almost all diurnal fixes and about two‐thirds of nocturnal ones were located in the gravel riverbed, which not only provided suitable breeding territories but probably part of the food resources needed for reproduction. Nocturnal excursions from breeding sites to feeding areas (mostly farmland), sometimes of a few kilometres, indicated that these resources do not cover all of the birds’ needs. Night spotlight counts of foraging birds in the agricultural area indicated that Stone‐curlews preferred recently harvested crops (mainly forage and wheat) and piles of farmyard manure. These habitat preferences are closely linked to the predominant agricultural activity of the study area, which is characterized by a high density of dairy farms for the production of Parmigiano Reggiano cheese. It seems likely that the proximity of gravel riverbed and farmland habitats is one of the main causes of the high breeding density recorded in the study area. We propose that the conservation of Stone‐curlews at this site could potentially be achieved only by a synergistic management of both natural and agricultural habitats.