Eggs of the tropical species A. (Oc.) epactius reared under SD were wider than those reared under LD. Electron microscopy studies of eggs of the close temperate species A. (Oc.) atropalpus able of diapause revealed different and stronger modifications in size and shape: LD eggs were longer and narrower than SD eggs, with changes

in the outer chorion structure ( Linley and Craig, 1994). However no differentiation of the possible factors, day length and diapause, responsible for these changes was obtained. Our study is thus the first to demonstrate that maternal photoperiod, and not diapause, influences egg volume in an Aedes species capable of diapause. The structure of XL184 cost mosquito eggs is therefore sensitive to several seasonal factors. Indeed, Anopheles sacharovi (Favre) and Anophelespunctipennis (Say) produce “winter” eggs almost totally covered by exochorion ( Theodor, 1925 and Fritz and Washino, 1992), and “winter” eggs of A. sacharovi possess a small float and are larger than “summer” float-less eggs. In these cases, the morphological differentiation originates in response

to temperature fluctuations, and not from the diapause syndrome, as diapause occurs at the larval or adult stages in Anopheles species ( Theodor, 1925). The latter are capable of egg quiescence, a process fairly similar to diapause at the molecular level ( Poelchau et al., 2013b), however quiescence is Dinaciclib mouse by definition an aseasonal state of inactivity ( Vinogradova, 2007). The mechanisms involved in Isotretinoin egg structure variability in mosquito are not determined and may be

multiple. Concerning the photoperiodic causality, we suspect that a circadian rhythm plays a part in the hormonal production and reserve storage, such as was demonstrated in several insect groups, including mosquitoes (Bloch et al., 2013). Egg production is regulated by hormones which are photophase dependent, as demonstrated in Hemiptera Rhodnius prolixus ( Vafopoulou et al., 2012). Lipids represent the major energetic source of eggs and are essential for the development of the embryo. Lipid reserve in eggs is provided by the mother ( Ziegler and Van Antwerpen, 2006). If that storage is dependent of photoperiod, and is more particularly developed during scotophase, long nights will enhance egg volume. Organism size cannot be explained by the simple sum of mechanisms that regulate the size and number of cells in organs ( Nijhout, 2003), but a positive relationship exists with the energy stock and egg size in some species, like the butterfly Bicyclus anynana ( Geister et al., 2009). A study carried out on a US temperate strain of A. albopictus found a lipid reserve more important by 30% in diapause-induced pharate larvae ( Reynolds et al., 2012), linked to an increase in egg volume.

Monkeys with medial, but not lateral, OFC lesions also exhibit irrational context-dependence of their choices in a 3-option probabilistic decision making task; after surgery, logistic functions describing the pattern of choices between pairs of options became affected by the value of the 3rd available option in these animals, violating normative models of rational choice [29•]. Such effects were particularly prominent during difficult choices. What is common to situations that recruit or require VMPFC during value-guided decision making is that, first, the goal is clearly selectable from currently this website presented stimuli and, second,

the task environment requires relevant information to be sampled and selected

for an optimal choice to be made. Indeed, an alternative account of the chosen minus unchosen comparison signal in VMPFC is that it instead reflects the difference between an attended and an unattended option, especially as chosen items generally are attended longer than unchosen ones 46 and 47•]. Neurons in dorsal parts of VMPFC encode value information particularly around attentional shifts, suggesting integration between the allocation of attention and valuation processes [48]. A change in the way information is attended to and acquired following VMPFC damage [49] might explain why the predominant deficit observed experimentally Orotic acid in monkeys Ferroptosis activation and humans with VMPFC damage is an increased tendency for inconsistent choices 15•, 50 and 51]. Unlike the maladaptive increase in exploratory choices seen following OFC lesions [28], this cannot be explained by impaired value learning [29•]. One way of integrating these ideas is to suggest that VMPFC does not just mediate value comparison, but is also required to maintain selective focus on information that is most relevant to the current goal. Chau and colleagues [52••] investigated how the presence of a third, but unavailable and therefore irrelevant, alternative would influence speeded choices between two other relevant options (Figure 2A). They found that

people would on average make more suboptimal choices during difficult decisions when the value of the unavailable distractor was comparatively low and the presence of such a low value distractor reduced the VMPFC value comparison signal (Figure 2B-C). Moreover, subjects who showed the greatest influence of the distractor on the VMPFC value comparison signal also made fewer choices of the best option (Figure 2D). There was also evidence that this process was influenced by interactions with OFC. The value comparison signal in VMPFC was positively coupled with activity in lateral OFC whereas the influence of the distractor on the VMPFC signal was negatively coupled with a similar part of lateral OFC.

Since our inception, both the physiotherapy profession and the MACP have both moved on considerably. Manipulation is now taught as an undergraduate skill and is well established within usual physiotherapy practice. It is one of many tools used to treat neuro-musculoskeletal disorders, and

is still an important technique in the tool bag of techniques available http://www.selleckchem.com/products/Bortezomib.html to us. We have all moved forward in our understanding of the interaction of the bio-psycho and social on patient outcomes, and our practice has developed accordingly. The new name of the MACP helps to reflect this broader view of our approach to managing people with musculoskeletal disorders. The proposed name change follows an extended period of consultation and discussion with members over the last 2 years or so, and is driven by members desire to have a name that reflects the breadth of the skills and experience within the organisation. We are very happy to head into the

future with our new name, but our old acronym, and can assure everyone that we will strive to maintain selleck kinase inhibitor the highest standards set by our visionary predecessors. “
“The authors of the above paper regret that there was an error concerning the scale of the Neck Disability Index (NDI). The correct scale is from 0 (No disability) to 100 (Maximum disability), instead of 0 to 50. The errors can be found in the following sections: 2.6.2. Prognostic and clinical variables “
“The four rotator cuff muscles not only move but also stabilize the glenohumeral joint by centralizing the humeral head in the glenoid fossa Thiamet G (Neri et al., 2009). Tears of the rotator cuff tendons may cause shoulder pain and can limit shoulder

function. Also in asymptomatic shoulders a rotator cuff tear (RotCuffTear) can be present. It was found in 23% of those with asymptomatic shoulders (n > 400, >50 years) ( Tempelhof et al., 1999). It is known that the prevalence of RotCuffTears increases with age and is more frequently reported in males ( Milgrom et al., 1995, Tempelhof et al., 1999 and Yamamoto et al., 2010). Genetic influences may also play a role ( Gwilym et al., 2009). In a recent systematic review, no associations were found between jobs or risk factors and the occurrence of RotCuffTears ( Van Rijn et al., 2010). Therefore, it remains unclear which conditions convert an asymptomatic RotCuffTear into a painful symptomatic tear. On the basis of imaging findings alone, it is impossible to differentiate between RotCuffTears leading to clinical symptoms and those without symptoms ( Schibany et al., 2004). It is suggested that the location rather than the size of the tear plays an important role ( Burkhart, 1991 and Burkhart et al., 1994). Although other shoulder muscles can compensate for the cuff tear, the critical amount of intact tendon or muscle necessary to maintain normal strength and normal range of motion has not yet been defined ( Schibany et al., 2004).

Moreover, hearing loss was diagnosed. The external genitalia were normal. Further examinations

at the age of click here 3 and 5 months showed normal psychomotor and somatic development (Fig. 1a). DNA was isolated from peripheral blood leukocytes of the patient and his healthy parents. Exons 1 through 27 of TCOF1, including exon–intron borders, were amplified by PCR under optimal conditions, using specific primers. The PCR products were subjected to multitemperature single-stranded conformation polymorphism (MSSCP) analysis at 5 °C, 15 °C and 25 °C, using the DNA Pointer Mutation Detection System. The electrophoresis was followed by silver staining. The PCR products were purified on the DNA GelOut columns (A&A Biotechnology, Poland) followed by direct sequencing with the use of a BigDye ver.3.0 dye terminator cycle sequencing kit and specific primers. The dideoxy-terminated fragments were identified by capillary gel-electrophoresis based on the ABI 310 DNA Analysis System. The MSSCP analysis of the amplified fragments of exon 13 of the TCOF1 gene demonstrated changes

in the electrophoretic mobility in this patient, while the changes were not observed in the patient’s parents. selleck compound In order to confirm the results obtained in MSSCP analysis a direct sequence analysis was performed. Sequence analysis demonstrated a novel, heterozygotic c.1978delC mutation in exon 13 of TCOF1. In the case of the patient’s parents direct DNA sequencing showed normal sequences ( Fig. 1b). A majority of mutations responsible for Treacher Collins syndrome are localized in exons, mainly in the hot spots in exons 10, 13, 15, 16, 23 and 24 [9]. The most commonly occurring mutations of the TCOF1 gene include deletions, which cause a shift of the reading frame, formation of the termination codon and shortening

of the protein Thiamine-diphosphate kinase product. The next most common mutations of the TCOF1 gene are insertions, the longest insertion localized on exon 5 [14]. In the presented patient a novel, heterozygotic deletion c.1978delC was detected in the TCOF1 gene. This mutation was absent in the patient’s parents which probably indicates a de novo origin. Analysis of the novel c.1978delC deletion with the use of the OMIGA 2.0 system indicates that it causes a premature termination of translation at 677aa, which results in the formation of a protein product of the gene devoid of the nuclear localization signal. We believe that these findings will facilitate precise diagnosis of the patient and will extend our knowledge on the pathogenesis of TCS. Molecular diagnosis of TCS is essential in prenatal and postnatal screening, being of great importance for genetic counseling as well. BAM-K – study design, data collection and interpretation, acceptance of final manuscript version, literature search. RS – data collection, acceptance of final manuscript version. MMS – acceptance of final manuscript version. None declared. None declared.

Jesußek et al. (2012) showed with their column experiments that temperatures of 25 °C and above lead to the mobilization of organic carbon and an increase in microbial activity. The increased availability of organic carbon combined with a higher microbial activity causes the redox zoning to shift toward more reducing conditions. Since the occurrence and rate of nitrate, iron and sulfate reduction are dependent on the redox conditions a temperature increase can have a strong influence on these processes. The findings of this study predict that at temperatures of 25 °C and higher, the usability of groundwater as drinking and process water can be impaired by reducing metal oxides and thus possibly releasing heavy metals from

the sediment. The column experiments performed by Bonte et al., this website 2013a and Bonte et Trametinib mouse al., 2013b showed that water quality was not affected when anoxic aquifer sediments were subjected to lower temperature (5 °C) than in situ temperature (11 °C). But at 25 °C, the concentration of As was significantly increased and at 60 °C also significant effects on the pH, dissolved organic carbon (DOC), P, K, Si, Mo, V, B and F were observed. The same experimental setup was used to determine the effect of temperature variations (5–80 °C) on redox processes and associated microbial communities (Bonte et al., 2013a). Both the hydrochemical and microbiological

data showed that a temperature increase from the in situ 11 °C to 25 °C caused a shift from iron-reducing to sulfate-reducing and methanogenic conditions. A further temperature increase to more than 45 °C resulted in the emergence of a thermophilic microbial community specialized in fermentation and sulfate reduction. Natural or contaminant organic components in groundwater can adsorb to sedimentary components, in particular organic material. In addition, groundwater composition is influenced by cation-exchange

on clay minerals and oxides. A hydrogeochemical reactive transport model (PHREEQC) using the results from previously described column experiments (Bonte et al., 2013a and Bonte et al., 2013b) revealed that sorption of anions decreases with temperature whereas sorption of cations increases with temperature (Bonte, 2013). Bumetanide Results showed that As and B are desorbed in the center of the warm water plume and mobilized toward the fringe of the warm water plume and the center of the cold water plume where these solutes become resorbed. According to Chiang et al. (2001), sorption of chlorinated methanes (carbon tetrachloride (CCl4), chloroform (CHCl3), methylene chloride (CH2Cl2)) also depends on temperature. Sorption of these VOCs decreases with increasing temperature. From about 8–16 °C, this decrease is about 10%. Since cation-exchange in aquifers takes place competitively on clay minerals, oxides and organic matter, each with other exchange properties, the derivation of thermodynamic constants per cation is difficult.

While the inter-assay CVs were acceptable, future improvements in reproducibility may be achieved with the development of rigorous assay-to-assay normalization controls and with better mixing approaches for the large and relatively dense 240 micron glass beads (cylinders), which tend to settle quickly and may result in poor and inconsistent mixing and binding kinetics. Likewise, the VeraCode™ system was also technically validated against ELISA selleck kinase inhibitor for detection of non-antibody circulating protein biomarkers using a sandwich immunoassay format. In this case, the CRC biomarker CEA was used as a model system. Here, 94% hit concordance was seen between the two assay types in 52 CRC samples (and quantitative correlation of

R2 = 0.9 when a linear regression is performed between the assays). Not surprisingly, the only discordant hits were borderline positive or negative CRC samples that fell extremely close to the cutoffs (see red asterisks in Fig. 3A), as the consistently low background click here in the normal patients resulted in a very low scoring

cutoff (both assays show 100% specificity against normal samples). Next, by combining the most robust TAA observed in our studies, p53, with sandwich immunoassay based quantification of the well-known CRC biomarker CEA, and the cytokine GDF15 in a hybrid multiplexed assay, we achieved a composite diagnostic sensitivity and specificity of 54% and 98%, respectively (186 samples CRC and normal). Thus, we demonstrate the ability to measure, in multiplex, two distinctly different biomarker types using different assay formats, simultaneously, on the VeraCode™ beads. As with the TAAs alone, the additive benefit of combing multiple biomarkers stems from the lack of complete redundancy, with each biomarker detecting several patients (9 to 29) which the others ADP ribosylation factor did not, and with no single biomarker exceeding 38% sensitivity (GDF15). It is important to emphasize that while the particular biomarkers used here were chosen to exemplify the immunoassay method, the clinical studies

performed here were only preliminary, retrospective validation studies on a particular cohort of CRC and normal patient samples, and that the results of these studies would need further validation using larger patient cohorts, as well as non-target disease controls (e.g. inflammatory bowel disease and cancers other than CRC) and ultimately, blinded studies and prospective clinical studies. In the future, it is expected that the CRC biomarker panel not only would expand, but also would be refined through elimination of biomarkers as further studies are performed using the VeraCode™ immunoassay methods presented here. For example, GDF15 is a stress-induced cytokine and in addition to CRC has been shown to be a biomarker for a variety of conditions such as heart disease (reviewed in Wollert and Kempf, 2012) and worsening albuminuria in patients with type 2 diabetes (Hellemons et al., 2012).

We will represent the visible layer activation variables by v  i, the hidden activations by h  j and the vector variables by v=viv=vi and h=hjh=hj where i=[1‥N]i=[1‥N] and j=[1‥S]j=[1‥S] index the individual neurons in the visible and hidden layers, respectively. Restricted Boltzmann Machines   are stochastic models that assume symmetric connectivity between the visible and hidden layers (see Fig. 1A) and seek to model the structure of a given dataset. They are energy-based models,

where the energy of a given configuration of activations vivi and hjhj is given by ERBM(v,h|W,bv,bh)=−v⊤Wh−bv⊤v−bh⊤h,and the probability of a given configuration is given by P(v,h)=exp(−ERBM(v,h|W,bv,bh))/Z(W,bv,bh),where Z(W,bv,bh)Z(W,bv,bh) is the partition function. One can extend the

RBM to continuous-valued Sotrastaurin datasheet visible variables by modifying the energy function, to obtain the Gaussian-binary RBM ERBM(v,h|W,bv,bh)=−v⊤σ2Wh+∥bv−v∥22σ2−bh⊤h.RBMs are usually trained through contrastive divergence, which approximately follows the gradient of the cost function CDn(W,bv,bh))=KL(P0(v|W,bv,bh)||P(v|W,bv,bh))−KL(Pn(v|W,bv,bh)||P(v|W,bv,bh)),CDn(W,bv,bh))=KL(P0(v|W,bv,bh)||P(v|W,bv,bh))−KL(Pn(v|W,bv,bh)||P(v|W,bv,bh)),where ABT-263 P  0 is the data distribution and P  n is the distribution of the visible layer after n   MCMC steps ( Carreira-Perpinan and Hinton, 2005). The function CD  n gives an approximation to maximum-likelihood (ML) estimation of the weight matrix ww. Maximizing the marginal probability P(vD|W,bv,bh)P(vD|W,bv,bh) of the data vDvD in the model leads to a ML-estimate which is hard to compute, as it involves averages over the equilibrium distribution P(v|W,bv,bh)P(v|W,bv,bh). The parameter update for

an RBM using CD learning is then given by Δθ∝〈∂ERBM∂θ〉0−〈∂ERBM∂θ〉n,where the <>n<>n denotes an average over the distribution Pn of the hidden and visible variables after n MCMC steps. The Cobimetinib weight updates then become ΔWi,j∝1σ2〈vihj〉0−1σ2〈vihj〉n.In general, n=1 already gives good results ( Hinton and Salakhutdinov, 2006). Autoencoders   are deterministic models with two weight matrices W1W1 and W2W2 representing the flow of data from the visible-to-hidden and hidden-to-visible layers, respectively (see Fig. 1B). AEs are trained to perform optimal reconstruction of the visible layer, often by minimizing the mean-squared error (MSE) in a reconstruction task. This is usually evaluated as follows: Given an activation pattern in the visible layer vv, we evaluate the activation of the hidden layer by h=sigm(v⊤W1+bh)h=sigm(v⊤W1+bh), where we will denote the bias in the hidden layer by bhbh. These activations are then propagated back to the visible layer through v^=sigm(h⊤W2+bv) and the weights W1W1 and W2W2 are trained to minimize the distance measure between the original and reconstructed visible layers.

NADPH molar extinction coefficient of 6.22 mM−1 cm−1 was utilized for enzyme activity

calculation ( Glock and Mclean, 1953). Cells cultured in 24-well plates were washed with PBS and frozen at −76 °C. Cell lysate was suspended in 100 μl of ice-cold PBS, and 200 μl of lysate suspension (from two wells) was transferred to 1.5 ml tubes and centrifuged at 10,000g for 10 min at 4 °C. A volume of 50 μl of the supernatant was separated for protein determination and the remaining volume (150 μl) was mixed with 30 μl of 50% trichloroacetic acid for protein precipitation after centrifugation at 10,000g (10 min, 4 °C). Then, 150 μl of this supernatant was transferred to a new tube and the pH was neutralized this website with 390 μl of Tris (0.4 M, pH 8.9). Finally, 200 μl of the neutralized solution was separated in two tubes of 1.5 ml for quantification of total glutathione and reduced glutathione, respectively. In the first tube, 26 μl of solution containing 0.9 U ml−1 of glutathione disulfide find more reductase and 1.8 mM of NADPH was added; the second tube received the 26 μl of Tris buffer (0.4 M, pH 8.9). After 10 min incubation at room temperature, 200 μl of tubes’ content were added to a 96-well microplate. Finally, 20 μl DTNB solution (2.5 mM of 5,5′-dithiobis

(2-nitrobenzoic acid) in 25% methanol) were added to microplate wells, and after 5 min, the absorbance was measured at 415 nm. GSH concentration was calculated by comparison with a standard curve of GSH ( Sedlak and Lindsay,

1968 and Griffith, 1980 with modifications). Glutathione disulfide concentration was calculated through the difference between total glutathione and GSH. Cells cultured in 24-well plates were washed with PBS and frozen at −76 °C. Cell lysate were suspended with 300 μl of ice-cold PBS, transferred to a 1.5 ml tube and centrifuged (12,000g, 20 min, 4 °C). A volume of 200 μl of supernatant was transferred Avelestat (AZD9668) to a 2.0 ml tube and mixed with 500 μl of DNPH solution (10 mM of 2,4-dinitrophenylhydrazine in 2.0 M of hydrochloric acid). For the blank, 2.0 M hydrochloric acid (without DNPH) was utilized. Samples were incubated at 30 °C during 90 min, proteins were precipitated with 1.0 ml of 28% trichloroacetic acid and centrifugation at 9000g for 10 min, and pelleted proteins were washed three times by suspension in ethanol/ethyl acetate (1:1) followed by centrifugation. Proteins were solubilized in 6.0 M of guanidine hydrochloride and tubes were centrifuged at 9000g for 5 min to remove any trace of insoluble material. The carbonyl content was determined spectrophotometrically at 360 nm using the molar absorption coefficient of 2.1 × 104 M−1 cm−1 for hydrazones and normalized by total protein content quantified in an aliquot reserved from the first centrifugation procedure ( Levine et al., 1994 and Quinlan and Gutteridge, 2000). Cells cultured in 24-well plates were washed with PBS and frozen at −76 °C.

Studies wherein the primary outcome variable is fasting TG level are challenging for several reasons. Serum TG levels are known to show day-to-day biological variations within individuals that can be as high as 25% in healthy selleck products fasted subjects when measured 2.5 months apart [24]. Hypertriglyceridemic individuals can have even greater fluctuations in fasting TG levels. Other reasons for intra-individual variability in TG measures can be associated with the preparation, processing, storage, and analysis

of blood samples. Despite attempts to minimize variability during sample collection, storage, shipment, and measurement, the individual biological fluctuations in fasting TGs were large, thereby resulting in a much higher intra-individual variation than accounted for in the power calculation (Supplementary Fig. 1). Multiple TG measurements at the individual visits, higher subject numbers or less dose groups check details should be considered in future studies. In order to circumvent these

limitations, an explorative data analysis approach was chosen to increase the statistical power of the study. Hence, the mean of 6 and 12 weeks treatment TG measurements of the four krill oil groups were pooled in a group- and time-independent manner. Across the 4 krill oil groups, the mean intake of krill oil was 1.875 g/day, and the associated intake of EPA and DHA was calculated to be 385 mg/day. This theoretical intake of EPA and DHA resulted in a 6.3% reduction from baseline in fasting TGs and a 10.2% placebo-adjusted reduction from baseline in fasting TGs. The efficacy of krill oil in reducing fasting serum TG levels has been reported in other studies; however, the doses of krill oil administered were larger than what was administered in the current study. Ulven et al. demonstrated that a daily dose

of 2 g krill oil lowered fasting TGs in participants with borderline high and high TG levels over a 7-week period [25]. Krill oil has also been found to be effective in hyperlipidemic patients without exclusion of lipid-lowering medication by significantly reducing total cholesterol, LDL-C, and TG, and by increasing HDL-C levels after 3 months L-NAME HCl of supplementation; moreover, krill oil appeared more effective than fish oil in reducing glucose, TG, and LDL-C levels [26]. The study, however, lacked information about the nature of the placebo and, more importantly, information about the baseline characteristics of the groups, particularly with respect to medication use (i.e. lipid-lowering drugs). Very recently, a pilot study demonstrated that daily supplementation of 4 g krill powder (containing 60% krill oil) over 24 weeks showed a significant TG-lowering effect in obese subjects [27].

Seedlings of each cultivar were then

exposed to different N deficiency stress treatments at the five-leaf stage. Hoagland’s solution without N [Ca(NO3)2·4H2O] was then added to maintain various N deficiency treatments [20], including mild stress [N2: 1.5 mmol L− 1 Ca(NO3)2·4H2O], moderate stress (N1: 0.15 mmol L− 1), extreme stress (N0: 0 mmol L− 1) Selleckchem Antiinfection Compound Library and a stress-free control (full strength Hoagland’s nutrient solution, modified). The solutions were refreshed twice a week and the pH of the nutrient solutions was adjusted to 5.5–6.5 every 2 days. An air pump was used for ventilation 24 h per day. Agronomic and physiological traits were evaluated 60 days after treatment. Sixty days after treatments, the tiller number, height (from the pot surface to the end of the longest leaf on the tallest tiller), aboveground biomass, leaf area, and root area were measured. Aboveground biomass was cut at the pot surface and separated into shoots and leaves, the leaf area was determined HDAC inhibitors list with a LI-COR 3100 leaf area meter (Li-Cor, Lincoln, NE) and the root surface area was determined with a root scanner (Epson Expression 1000XL, Japan). Roots and rhizomes were washed free of growth media and all plant samples were treated at 105 °C for 30 min

for fixation and then oven dried at 65 °C until a constant weight was reached. The presence of rhizomes was recorded and the root to shoot weight ratio (R:S) was calculated. Gas exchange measurements were performed two weeks after treatment initiation using a portable open gas exchange system (LI-6400, LI-COR) calibrated to deliver a photosynthetic FER photon flux density of 2000 μmol m− 2 s− 1 and an ambient CO2 of 400 μmol mol− 1 (supplied by a LI-COR CO2 injector) and a leaf temperature of (30 ± 1) °C. Data were collected for 2 min at 5-s intervals for three randomly chosen plants from each treatment listed above (eight replications per treatment) on the youngest fully expanded leaf on the longest tiller, as described by Barney et

al. [12]. Net CO2 assimilation (A), transpiration (E), and stomatal conductance (gs) were recorded, and photosynthetic water use efficiency (WUE) was calculated (WUE = net photosynthesis/transpiration). Chlorophyll a and chlorophyll b were extracted with 80% acetone from the same leaf as used for gas exchange measurements. Absorbance was measured at 663 nm and 645 nm for chlorophyll a and chlorophyll b, respectively, using a UV spectrophotometer (UV-2550, Shimadzu). Total chlorophyll content was calculated according to the procedure described by Lichtenthaler and Wellburn [21]. To avoid the negative influence of different cultivars on the evaluation of tolerance, the Low-N tolerance index (LNT) was calculated. This is the ratio of the index under treatment to that of the control (LNT = (value of tested traits under treatments/value of same tested traits under control) × 100%).