Root water uptake depth determines the hydraulic vulnerability of temperate European tree species during the extreme 2018 drought.

We took advantage of the European 2018 drought and assessed the mechanisms causing differences in drought vulnerability among mature individuals of nine co-occurring tree species at the Swiss Canopy Crane II site in Switzerland. Throughout the drought we monitored leaf water status and determined native embolism formation in the canopy of the trees as indicators of drought vulnerability. We also determined hydraulic vulnerability thresholds (Ψ12 -, Ψ50 - and Ψ88 -values), corresponding hydraulic safety margins (HSMs) and carbohydrate reserves for all species as well as total average leaf area per tree, and used stable isotopes to assess differences in root water uptake depth among the nine species as variables predicting differences in drought vulnerability among species. Marked differences in drought vulnerability were observed among the nine tree species. Six species maintained their water potentials above hydraulic thresholds, while three species, Fagus sylvatica, Carpinus betulus and Picea abies, were pushed beyond their hydraulic thresholds and showed loss of hydraulic conductivity in their canopies at the end of the drought. Embolism resistance thresholds and associated HSMs did not explain why the co-existing species differed in their drought vulnerability, neither did their degree of isohydry, nor their regulation of carbohydrate reserves. Instead, differences in structural-morphological traits, in particular root water uptake depth, were associated with the risk of reaching hydraulic vulnerability thresholds and embolism formation among the nine species. Our study shows that structural-morphological traits, such as root water uptake depth, determine how quickly different species approach hydraulic vulnerability thresholds during a drought event and can thus explain species differences in drought vulnerability among mature field-grown trees.

A systematic review of surgical site infections following day surgery: a frequentist and a Bayesian meta-analysis of prevalence.

BACKGROUND: Since 1990, several studies have focused on safety and patient satisfaction in connection with day surgery. However, to date, no meta-analysis has investigated the overall prevalence of surgical site infections (SSI). AIM: To estimate the overall prevalence of SSI following day surgery, regardless of the type of surgery. METHOD: A systematic review and a meta-analysis of the prevalence of SSI following day surgery, regardless of the type of surgery, was conducted, seeking all studies before June 2016. A pooled random effects model using the DerSimonian and Laird approach was used to estimate overall prevalence. A double arcsine transformation was used to stabilize the variance of proportions. After performing a sensitivity analysis to validate the robustness of the method, univariate and multi-variate meta-regressions were used to test the effect of date of publication, country of study, study population, type of specialty, contamination class, time of postoperative patient visit after day surgery, and duration of hospital care. FINDINGS: Ninety articles, both observational and randomized, were analysed. The estimated overall prevalence of SSI among patients who underwent day surgery was 1.36% (95% confidence interval 1.1-1.6), with a Bayesian probability between 1 and 2% of 96.5%. The date of publication was associated with the prevalence of SSI (coefficient -0.001, P = 0.04), and the specialty (digestive vs non-digestive surgery) tended to be associated with the prevalence of SSI (coefficient 0.03, P = 0.064). CONCLUSION: The meta-analysis showed a low prevalence of SSI following day surgery, regardless of the surgical procedure.

Small-scale patterns in snowmelt timing affect gene flow and the distribution of genetic diversity in the alpine dwarf shrub Salix herbacea.

Current threats to biodiversity, such as climate change, are thought to alter the within-species genetic diversity among microhabitats in highly heterogeneous alpine environments. Assessing the spatial organization and dynamics of genetic diversity within species can help to predict the responses of organisms to environmental change. In this study, we evaluated whether small-scale heterogeneity in snowmelt timing restricts gene flow between microhabitats in the common long-lived dwarf shrub Salix herbacea L. We surveyed 273 genets across 12 early- and late-snowmelt sites (that is, ridges and snowbeds) in the Swiss Alps for phenological variation over 2 years and for genetic variation using seven SSR markers. Phenological differentiation triggered by differences in snowmelt timing did not correlate with genetic differentiation between microhabitats. On the contrary, extensive gene flow appeared to occur between microhabitats and slightly less extensively among adjacent mountains. However, ridges exhibited significantly lower levels of genetic diversity than snowbeds, and patterns of effective population size (Ne) and migration (Nem) between microhabitats were strongly asymmetric, with ridges acting as sources and snowbeds as sinks. As no recent genetic bottlenecks were detected in the studied sites, this asymmetry is likely to reflect current meta-population dynamics of the species dominated by gene flow via seeds rather than ancient re-colonization after the last glacial period. Overall, our results suggest that seed dispersal prevents snowmelt-driven genetic isolation, and snowbeds act as sinks of genetic diversity. We discuss the consequences of such small-scale variation in gene flow and diversity levels for population responses to climate change.

Increased spring freezing vulnerability for alpine shrubs under early snowmelt.

Alpine dwarf shrub communities are phenologically linked with snowmelt timing, so early spring exposure may increase risk of freezing damage during early development, and consequently reduce seasonal growth. We examined whether environmental factors (duration of snow cover, elevation) influenced size and the vulnerability of shrubs to spring freezing along elevational gradients and snow microhabitats by modelling the past frequency of spring freezing events. We sampled biomass and measured the size of Salix herbacea, Vaccinium myrtillus, Vaccinium uliginosum and Loiseleuria procumbens in late spring. Leaves were exposed to freezing temperatures to determine the temperature at which 50% of specimens are killed for each species and sampling site. By linking site snowmelt and temperatures to long-term climate measurements, we extrapolated the frequency of spring freezing events at each elevation, snow microhabitat and per species over 37 years. Snowmelt timing was significantly driven by microhabitat effects, but was independent of elevation. Shrub growth was neither enhanced nor reduced by earlier snowmelt, but decreased with elevation. Freezing resistance was strongly species dependent, and did not differ along the elevation or snowmelt gradient. Microclimate extrapolation suggested that potentially lethal freezing events (in May and June) occurred for three of the four species examined. Freezing events never occurred on late snow beds, and increased in frequency with earlier snowmelt and higher elevation. Extrapolated freezing events showed a slight, non-significant increase over the 37-year record. We suggest that earlier snowmelt does not enhance growth in four dominant alpine shrubs, but increases the risk of lethal spring freezing exposure for less freezing-resistant species.

Unrestricted quality of seeds in European broad-leaved tree species growing at the cold boundary of their distribution.

BACKGROUND AND AIMS: The low-temperature range limit of tree species may be determined by their ability to produce and disperse viable seeds. Biological processes such as flowering, pollen transfer, pollen tube growth, fertilization, embryogenesis and seed maturation are expected to be affected by cold temperatures. The aim of this study was to assess the quality of seeds of nine broad-leaved tree species close to their elevational limit. METHODS: We studied nine, mostly widely distributed, European broad-leaved tree species in the genera Acer, Fagus, Fraxinus, Ilex, Laburnum, Quercus, Sorbus and Tilia. For each species, seeds were collected from stands close to optimal growth conditions (low elevation) and from marginal stands (highest elevation), replicated in two regions in the Swiss Alps. Measurements included seed weight, seed size, storage tissue quality, seed viability and germination success. KEY RESULTS: All species examined produced a lot of viable seeds at their current high-elevation range limit during a summer ranked 'normal' by long-term temperature records. Low- and high-elevation seed sources showed hardly any trait differences. The concentration of non-structural carbohydrates tended to be higher at high elevation. Additionally, in one species, Sorbus aucuparia, all measured traits showed significantly higher seed quality in high-elevation seed sources. CONCLUSIONS: For the broad-leaved tree taxa studied, the results are not in agreement with the hypothesis of reduced quality of seeds in trees at their high-elevation range limits. Under the current climatic conditions, seed quality does not constitute a serious constraint in the reproduction of these broad-leaved tree species at their high-elevation limit.

Detection and differentiation of sensorineural hearing loss in mice using auditory steady-state responses and transient auditory brainstem responses.

Sensorineural hearing loss (SNHL) comprises hearing disorders with diverse pathologies of the inner ear and the auditory nerve. To date, an unambiguous phenotypical characterization of the specific pathologies in an affected individual remains impossible. Here, we evaluated the use of scalp-recorded auditory steady-state responses (ASSR) and transient auditory brainstem responses (ABR) for differentiating the disease mechanisms underlying sensorineural hearing loss in well-characterized mouse models. We first characterized the ASSR evoked by sinusoidally amplitude-modulated tones in wild-type mice. ASSR were robustly elicited within three ranges of modulation frequencies below 200 Hz, from 200 to 600 Hz and beyond 600 Hz in most recordings. Using phase information we estimated the apparent ASSR latency to be about 3 ms, suggesting generation in the auditory brainstem. Auditory thresholds obtained by automated and visual analysis of ASSR recordings were comparable to those found with tone-burst evoked ABR in the same mice. We then recorded ASSR and ABR from mouse mutants bearing defects of either outer hair cell amplification (KCNQ4-knockout) or inner hair cell synaptic transmission (Bassoon-mutant). Both mutants showed an increase of ASSR and ABR thresholds of approximately 40 dB versus wild-type when investigated at 8 weeks of age. Mice with defective amplification displayed a steep rise of ASSR and ABR amplitudes with increasing sound intensity, presumably reflecting a strong recruitment of synchronously activated neural elements beyond threshold. In contrast, the amplitudes of ASSR and ABR responses of mice with impaired synaptic transmission grew very little with sound intensity. In summary, ASSR allow for a rapid, objective and frequency-specific hearing assessment and together with ABR and otoacoustic emissions can contribute to the differential diagnosis of SNHL.

13C labelling reveals different contributions of photoassimilates from infructescences for fruiting in two temperate forest tree species.

The pathways of currently fixed carbon in fruit bearing branchlets were investigated in two temperate forest tree species (CARPINUS BETULUS and FAGUS SYLVATICA), which differ in texture of their vegetative infructescence tissues (leaf-like in CARPINUS vs. woody in FAGUS). During late spring, (13)C pulse-labelling was conducted on girdled, defoliated, girdled plus defoliated and untreated fruiting branchlets of mature trees IN SITU, to assess changes in C relations in response to the introduced C source-sink imbalances. At harvest in early August, 75 - 100 % of the recovered (13)C label was bound to infructescences (either fruits or vegetative infructescence tissue), revealing them as the prime C sinks for current photoassimilates. Leaves on girdled branchlets were not stronger labelled than on ungirdled ones in both species, indicating no upregulation of the leaves' photosynthetic capacity in response to the prevention of phloemic transport, which was also supported by measurements of light saturated photosynthesis. In contrast, (13)C labels tended to be higher after complete defoliation in the vegetative infructescence tissues of CARPINUS, suggesting enhanced net photosynthesis of green infructescence parts as compensation for the loss of regular leaves. The total labelling-derived (13)C content of whole infructescences was very similar between foliated and defoliated CARPINUS branchlets. Cupulae of FAGUS, on the other hand, remained almost unlabelled on defoliated branchlets, indicating the photosynthetic inactivity of this woody infructescence tissue. Consequently, CARPINUS still produced relatively high fruit masses on girdled plus defoliated branchlets, while in FAGUS fruit development ceased almost completely at this most severe treatment. Our results highlight that green vegetative infructescence tissue assimilates substantial amounts of C and can partly substitute regular leaves as C sources for successful fruit development.

Effects of Glyptapanteles liparidis (Hym.: Braconidae) parasitism, polydnavirus, and venom on development of microsporidia-infected and uninfected Lymantria dispar (Lep.: Lymantriidae) larvae.

Effects of parasitism, polydnavirus, and venom of the endoparasitoid Glyptapanteles liparidis on Lymantria dispar larvae infected with the microsporidium Vairimorpha sp. and uninfected hosts were studied. We tested the impact on growth and development of hosts, as well as on microsporidian infection. Both parasitism and polydnavirus/venom treatment alone caused a slight increase in growth rate and relative growth rate in uninfected fourth instar hosts. This effect was more pronounced with the addition of Vairimorpha infection. With no parasitism, however, infection reduced host growth markedly. Microsporidiosis delayed larval molts of L. dispar, and additional polydnavirus/venom treatment or parasitization induced significantly earlier molting. Polydnavirus/venom treatment of uninfected L. dispar resulted in prolonged larval development due to supernumerary molts and in higher pupal mortality. Infected larvae treated with polydnavirus/venom died earlier than infected larvae that were not treated and produced more Vairimorpha spores per unit fresh mass of the host.

Interactions between an entomopathogenic microsporidium and the endoparasitoid Glyptapanteles liparidis within their host, the gypsy moth larva.

Interactions in the host-parasitoid-pathogen system, Lymantria dispar L. (Lep., Lymantriidae)-Glyptapanteles liparidis (Bouché) (Hym., Braconidae)-Vairimorpha sp. (Protista, Microspora), were investigated. Host selection experiments revealed that G. liparidis females did not discriminate between infected and uninfected host larvae for oviposition. Transmission of the microsporidium from infected to uninfected hosts by stinging female wasps could not be ascertained. Females that developed in infected L. dispar larvae did not transmit the pathogen via oviposition. Vairimorpha infection of the host negatively affected the performance of the braconid, when inoculation took place either before or after parasitization. Microsporidiosis of the host caused delayed development, reduced pupation and adult eclosion, reduction in size and weight, and reduction of adult longevity of G. liparidis. Parasitoids themselves were not systemically infected by Vairimorpha sp., but braconid larvae did ingest microsporidian spores at the end of their endoparasitic development and accumulated the undigested and ungerminated spores in the blind midgut. Negative effects of host infection on parasitoid larvae were detectable from the beginning of parasitoid larval development. Lethal time was reduced when L. dispar larvae were infected and parasitized, often at the expense of the parasitoid when G. liparidis were unable to complete endoparasitic development before the host died. Intensity of infection, measured as number of spores produced per milligram fresh weight of L. dispar larva, was slightly higher in parasitized and infected hosts than in unparasitized and infected hosts.

Purification and characterization of stachyose synthase from lentil (Lens culinaris) seeds: galactopinitol and stachyose synthesis.

Stachyose synthase (STS) (EC 2.4.1.67) was purified 313-fold from mature seeds of lentil. The final preparation had a specific activity of 9.09 nkat stachyose formed per milligram of protein. The enzyme was a monomeric protein with a molecular mass of 88.6 kDa (SDS-PAGE) and an isoelectric point of 4.8 (chromatofocusing). Western analysis revealed cross-reactivity of polyclonal antibodies raised against STS from adzuki bean with the lentil enzyme. The purified enzyme catalyzed a range of different galactosyl transfer reactions. In addition to the genuine STS reaction (raffinose + galactinol --> stachyose + myo-inositol), the enzyme catalyzed the reversible galactosyl transfer from galactinol to d-pinitol (1d-3-O-methyl-chiro-inositol), yielding galactopinitol A (O-alpha-d-galactopyranosyl-(1 --> 2)-4-O-methyl-d-chiro-inositol) and myo-inositol. Galactopinitol A could be further galactosylated by STS to give ciceritol (O-alpha-d-galactopyranosyl-(1 --> 6)-O-alpha-d-galactopyranosyl-(1 --> 2)-4-O-methyl-d-chiro-inositol). Enzymatic synthesis of galactopinitol A and ciceritol is a new observation. However, STS was not only able to utilize galactopinitol A as galactosyl acceptor, but also as galactosyl donor to form stachyose from raffinose. The role of STS in the metabolism of galactosyl cyclitols and oligosaccharides in plant seeds is discussed.

The development of experimental autoimmune encephalomyelitis in the mouse requires alpha4-integrin but not alpha4beta7-integrin.

Because monoclonal antibodies (mAbs) directed against alpha4-integrin and VCAM-1 inhibit the development of experimental autoimmune encephalomyelitis (EAE) in vivo, it has been concluded that the successful therapeutic effect is due to interference with alpha4beta1/VCAM-1-mediated interaction of autoaggressive T cells with the blood-brain barrier. A possible role for alpha4beta7-integrin, or interference with other T cell mediated events during the pathogenesis of EAE, has not been considered. We have compared the effects of mAb therapy on the development of EAE in the SJL/N mouse, using a large panel of mAbs directed against alpha4, beta7, the alpha4beta7-heterodimer, and against VCAM-1. Although encephalitogenic T cells express both alpha4-integrins, mAbs directed against the alpha4beta7-heterodimer or against the beta7-subunit did not interfere with the development of EAE. In contrast, mAbs directed against alpha4 and VCAM-1 inhibited or diminished clinical or histopathological signs of EAE. Our data demonstrate for the first time that alpha4beta7 is not essential for the development of EAE. Furthermore, our in vitro studies suggest that the therapeutic effect of anti-alpha4-treatment of EAE might also be caused by inhibition of antigen-specific T cell proliferation.

T cell interaction with ICAM-1-deficient endothelium in vitro: essential role for ICAM-1 and ICAM-2 in transendothelial migration of T cells.

Transendothelial migration is a crucial step in the complex process of lymphocyte extravasation during lymphocyte homing, immunosurveillance and inflammation. However, little is known about the precise role of cell adhesion molecules (CAM) involved in this particular event. To define the CAM involved in T cell adhesion versus transendothelial migration, we have previously established an in vitro transendothelial migration system using mouse T cells and mouse endothelioma cells. We demonstrate here that, using ICAM-1-deficient endothelioma cells derived from ICAM-1 mutant mice, transendothelial migration of T cells was inhibited to a much greater extent when compared to migration across wild-type cells treated with a blocking anti-ICAM-1 monoclonal antibody. This unexpected result was confirmed by a rescue experiment using retroviral transfer of wild-type ICAM-1 into ICAM-1-deficient endothelial cells. Additional experiments showed that, in the absence of functional ICAM-1, only ICAM-2 was involved in transendothelial migration, but not PECAM-1, VCAM-1, or E-selectin. Taking this novel approach, we show that ICAM-1 and ICAM-2 are essential for transendothelial migration of T cells.

Immunosurveillance modelled in vitro: naive and memory T cells spontaneously migrate across unstimulated microvascular endothelium.

As a model for T cell immigration into non-lymphoid tissue we set up an in vitro assay that would allow us to investigate the phenotype of T lymphocytes from peripheral lymph nodes (PLN), mesenteric lymph nodes (MLN) or peripheral blood (PBL) of mice, which were able to spontaneously migrate across unstimulated microvascular endothelium. The transendothelial migrating T cell population was enriched for T lymphocytes expressing a "recently activated/memory' phenotype: LFA-1/CD44/ICAM-1high, but also contained CD45RBhigh and LFA-1low T cells, which in the case of MLN T cells were phenotyped as CD4+ and thus characterized as naive T cells. Transmigrated T cells could be further distinguished from their original populations and from each other by their distinct but heterogeneous expression patterns for L-selectin, alpha 4 beta 7-integrin and PECAM-1. This observation suggests the presence of phenotypically different migratory T cells among MLN, PLN and PBL. Additional studies provided evidence that the capacity to migrate across unstimulated microvascular endothelium was a characteristic of a T cell population that could phenotypically be differentiated from activated T cells. The endothelial cells were found to play an active role in selecting the traversing T cell population, as they controlled the number and phenotype of spontaneously transmigrating T cells. Our studies suggest that the capacity to transmigrate across unstimulated microvascular endothelium and hence to immigrate into non-lymphoid tissue is owned by a phenotypically heterogeneous T cell population, which is enriched for memory T cells but not devoid of naive T cells.

Midsagittal velar kinematics during production of VCV sequences.

We present data on velar speech movements examined by electromagnetic articulography (EMA), a tool for the study of velar kinematics. Based on investigations on 22 normal German subjects (15 male, 7 female), movement paths of the velum using a sensor positioned on the midsagittal anterior surface were recorded using a series of high and low vowel VCV utterances. Maximum velar elevation (mean = 13.5 +/- 0.6 mm [SD]) with the rest position as a reference was observed during the production /iti/. The average maximum articulatory velocity of velar movements during /iti/ production was 141.0 mm/sec. Maximum movement of the velum observed during production of /ana/ averaged 7.5 +/- 1.1 mm. The direction of velar movement ranged between 44.6 degrees and 50.4 degrees relative to the bite plane registered electromagnetically. The influence of loudness variation of sequence production is demonstrated and discussed with the literature.

Simultaneous evaluation of articulatory velopharyngeal function under different dynamic conditions with EMA and videoendoscopy.

Objective examination of velopharyngeal (VP) sphincter function has been described based on multiple methods. Videofluoroscopy and videoendoscopy represent today's clinical standard. A new concept of analysis of VP functioning is described using electromagnetic articulography (EMA) for midsagittal measurement of velar movement and videoendoscopy in a setting for simultaneous recording. Multidimensional analysis of VP functions using this setting is demonstrated for a normal subject. Timing aspects of VP closure and a transverse-sagittal ratio of VP port diameter are reported under different conditions of speech dynamics.

Electromagnetic articulography as a method for detecting the influence of spikes on tongue movement.

In orthodontics dysfunctions of the tongue are discussed as a factor in the development of malocclusion. A new objective method for examining tongue movement is the electromagnetic articulography (EMA). This enables movements of the tongue to be followed using the principle of inductive distance measurements. A helmet made of acrylic material serves as a carrier for three transmitter coils. These are located in front of the forehead, in front of the chin, and at the back of the neck. Receiver coils are fixed to the tip of the tongue and at two points on the dorsum of the tongue. The distance between the receiver and the transmitter coils is calculated by means of a personal computer. The movement trajectories are displayed on a computer screen in x-y-coordinates. Tongue movements of one patient with an anterior open bite are shown during characteristic test sounds and during swallowing. The first examination took place without any orthodontic appliance. In a second series in the same session, tongue movement was recorded under the influence of an orthodontic appliance with spikes. While the patient shows vertical combined with sagittal components in the movement pattern of the tongue without an orthodontic appliance, the same patient, under the influence of an orthodontic appliance with spikes, shows a change of the rest and work position of the tongue and in the pattern of tongue movement with a more posterior position of the tongue, and with an increase in the vertical and a decrease in the sagittal components of the movement pattern.

[Simultaneous electromagnetic articulography and video endoscopy. A case contribution to the objective diagnosis of the velopharyngeal sphincter]. / Simultane elektromagnetische Artikulographie und Videoendoskopie. Ein kasuistischer Beitrag zur objektiven Diagnostik des velopharyngealen Sphinkters.

An objective examination of velopharyngeal sphincter function (VP) is possible using a variety of signal and pictorial methods to differing degrees. Video endoscopy and videofluoroscopy are the currently accepted clinical standard. With the help of a new concept described here it is possible both to analyze VP function in the midsagittal plane using electromagnetic articulography (EMA) and also to examine simultaneously the horizontal plane video endoscopically. This multidimensional analysis of VP function is presented in a case study of a healthy test person and the timing aspects of VP closure and transverse-sagittal ratio of VP port-diameters ist reported.

Speech freezing in Parkinson's disease: a kinematic analysis of orofacial movements by means of electromagnetic articulography.

Kinematic recordings of orofacial movements by means of electromagnetic articulography were performed in an akinetic-rigid Parkinsonian patient presenting with intermittent speech freezing in diadochokinesis tasks (rapid repetitions of the syllable /ta/). During freezing periods the patient produced a sustained /a/ instead of the required consonant-vowel sequences. The underlying articulatory trajectories were characterized by repetition rates amounting to 8-10 Hz concomitant with reduced movement amplitudes. Obviously, the undershooting of articulatory gestures failed to establish a sufficient occlusion of the vocal tract giving rise to the perceived speech freezing. In contrast, preserved diadochokinesis occurred at frequencies of 4-6 Hz. Most probably, the increased articulatory repetition rate reflects a pacing of orofacial movements by released tremor oscillations. Due to slowing of articulatory movements spastic dysarthrics can also present with missing syllabic modulation during oral diadochokinesis. The results of the articulographic recordings demonstrate the different pathophysiology of Parkinsonian freezing.