0–Section F. The goal was to interview newly admitted residents within 24 hours of admission.

This would enable staff to address preferences from the beginning of the resident’s stay. Sites were asked to interview long stay residents shortly before the individual’s care planning conference. The next step was to conduct the Preference Satisfaction portion of the interview, ideally within 5 to 7 days after the initial preference interview for short stay residents. Long stay resident preference and satisfaction interviews could be see more conducted on the same day, or 5 to 7 days apart. Providers were given several options for the choice of interviewer for the preference and satisfaction portions of the interview. Guidelines recommended that the staff member who actually delivers the care should conduct the preference interview; however, to encourage residents to share forthright opinions, a different staff member could be assigned to ask preference satisfaction questions. Among the possible options, communities could (1) use a volunteer or personnel other than a certified nursing assistant (CNA) or activity therapist

to Epigenetic activity conduct preference satisfaction interviews; (2) have the CNA and activity therapist switch interview categories (ie, CNA asks questions about activity preferences, and activity therapist asks about personal care); or (3) deploy licensed nurses or social workers from a neighboring unit or floor to conduct preference satisfaction interviews.23 Staff from pilot sites entered responses from resident preference acetylcholine and satisfaction interviews into the revised Excel spreadsheet that automatically calculates a preference congruence percentage for each resident. Reports can be generated for each individual resident (for an example, Figure 1), or in aggregate for a household

of residents (Figure 2). As care planning conferences took place, staff members also noted whether the resident, family members or close friends and direct care staff, such as CNAs, attended the meetings and entered this data into the spreadsheet, which calculated participation rates. Pilot sites were asked to fax their NH’s 4 aggregate quality indicator results to the research team (for an example, Figure 3). Individual resident-level information was not shared with researchers. Project coordinators identified by each site were asked to complete a questionnaire (93 items) regarding staff experiences using the new toolkit. The evaluation form asked about the PCC spreadsheet’s functionality and content, the webinar training experience, the resident interview process, challenges in implementing PCC, and overall satisfaction with the toolkit. Responses for most questions used a 5-point Likert scale, with a range from “completely agree” to “completely disagree.” Also, several open-ended questions provided a qualitative perspective on these topics.

The membranes were then incubated for 2 h with anti-rabbit-HRP IgG for SYN, SYP, BDNF, GluR1 and GluR2/3 and 5-Fluoracil ic50 anti-mouse-HRP

IgG for NFs, MAP2 and GFAP (Amersham, Little Chalfont, Buckinghamshire, UK) diluted 1:10,000 in TTBS with 1% non-fat milk. The probed proteins were developed by using a chemiluminescent kit (ECL, Amersham Biosciences, NJ, USA). The membrane was then incubated for 30 min at room temperature with stripping buffer and an anti-β-actin antibody (Sigma, St. Louis, MO, USA) was used to quantify β-actin as a loading control. The bound antibodies were visualized using radiographic films which were placed in contact with the membranes, then developed and fixed. The quantification of band intensity was performed with Scion Image 4.0.2 (Scion Corporation, Frederick, MD, USA). The hippocampi were collected (8 animals per group) and immediately homogenized in 1 mL TRIzol (Invitrogen, Carlsbad,CA, USA) with a homogenizer and total RNA was isolated following the manufacturer’s suggested protocol. Briefly, following one chloroform extraction step, RNA was precipitated with isopropanol and the pellet washed once in 70% ethanol. After air-drying, RNA was resuspended in DEPC-treated

water and the concentration AZD6244 of each sample was obtained from A260/A280 nm measurements. Residual DNA was removed using DNase I (Invitrogen) by following the manufacturer’s protocol. For each 20 μL reverse transcription reaction, 4 μg total RNA was mixed with 1 μL oligodT primer (0.5 μg/μL; Invitrogen) and incubated for 10 min at 65 °C. After cooling on ice the solution was mixed with 4 μL 5× first strand buffer, 2 μL of 0.1 M DTT, 1 μL of dATP, dTTP, dCTP and dGTP (10 mM each), and 1 μL SuperScript III reverse transcriptase (200 U/μL; Invitrogen) and incubated for 60 min at 50 °C. Reaction was inactivated by heating at 70 °C for 15 min, and the samples were diluted four times. The real-time PCR reaction system included the following: 200 to 400 nM primers, Quinapyramine 5 ng cDNA samples, and 1× SYBR® Green PCR Master Mix (Applied Biosystems, Foster City, CA, USA). Using

the Rotor-Gene 3000 Real-time PCR detection system (Corbett Research, Mortlake, NSW, Australia), cycling conditions were set as follows: after initial activation at 50 °C for 2 min and 95 °C for 10 min, 40 cycles of 95 °C for 15 s and 60 °C for 1 min, then melt curve analysis was performed by heating samples from 65 °C to 99 °C (1 °C increment changes at 5 s intervals), in order to evaluate primer specificity. All sample measurements were performed in duplicate. Primers used for the housekeeping genes, hydroxymethylbilane synthase (HMBS) and hypoxanthine phosphoribosyltransferase 1 (HPRT1), were described by Depreter et al. (2002) and the primers for the genes of interest were designed using software primer express v3.0 (Applied Biosystems) (Depreter et al., 2002).

, 2002). Further research must be carried out in order to elucidate the mechanisms of anthocyanin Stem Cell Compound Library high throughput degradation during ohmic heating and confirm the hypothesis suggested in this work; future experiments should be conducted using lower voltages. A new system is being currently developed in our laboratory, which will allow us to evaluate lower voltages combined with different frequency ranges. This article presents a study concerning anthocyanin degradation during the thermal treatment of blueberry pulp using ohmic and conventional heating. For the ohmic heating experiments, the effects of the voltage and the solids content were evaluated. Most of the independent variables – quadratic and linear voltage

variables, the linear solids content variable and the interaction variable – had significant effects on the response values, the exception being the quadratic effect of the solids content. A second-order polynomial model was obtained, and the equation shows that anthocyanin degradation increases as both parameters analyzed increases. The level of degradation varied from 5.7 to 14.7% for the ohmic Anti-diabetic Compound Library purchase heating experiments, and for the conventional heating experiment, the level of degradation was 7.2%. The percentage of anthocyanin degradation was similar or even lower than those obtained with conventional heating when the ohmic heating process was used with low voltage gradients. When higher voltage gradients were applied,

the levels of degradation were greater for the ohmic-heated pulp. These results might be explained by electrochemical reactions that are catalyzed by high voltages. The results emphasize the importance of the use of inert materials in electrodes and electrode coatings or the use of high frequency power

to limit electrochemical reactions. The authors acknowledge the financial support received from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior). “
“Mangiferin (1,3,6,7-tetrahydroxy-2-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]xanthen-9-one) (MGN) (Fig. 1) is a naturally occurring polyphenol in several fruits, one being Mangifera indica L. (common name: mango), one of the most popular tropical fruit-bearing trees in the world ( Barreto et al., see more 2008). The interest in MGN stems from its wide range of biological actions, for instance, gastroprotective ( Carvalho et al., 2007), analgesic ( Dar et al., 2005), antibacterial ( Duang, Wang, Zhou, & Huang, 2011) together with cytoprotective ( Pardo-Andreu et al., 2006). The therapeutic potential of MGN has been investigated in the prevention and treatment of periodontitis ( Carvalho et al., 2009). A wide spectra of these properties have been attributed to its antioxidant properties, being MGN the major component (10–20%) of the aqueous formulation named Vimang® used in Cuba ( Garrido, González, Romay, Núñez-Sellés, & Delgado, 2008).

g. temperature, wind speed and direction). This data allows the application of the online simulation tool to get an impression at what time the pollution will reach a certain place (e.g. town, beach or harbour) and how it spreads in the river and in the lagoon. If the likely pollutant Smoothened antagonist (e.g. E. coli, Enterococci, viruses) is known, a more realistic

simulation is possible. It can take into account e.g. the die-off rates and the decay of problematic organisms and the potential pollutant concentrations at certain places can be estimated. If the authority comes to the conclusion that a risk exists, the simulation results allow to organize an optimized monitoring and to inform local actors when and where to take what kind of water sample. After the laboratory analysis, the data is stored and those locations where water quality thresholds are exceeded automatically receive an

alert email. On a regular and event-driven basis, bathing water quality data and other relevant information are distributed via newsletter to a broader public. The preparation and distribution are supported by a software tool. Our brief phone survey among several end-users showed that improved information about water quality aspects is appreciated. The newsletter structure and content where positively evaluated by users and above 25% planned to further disseminate it. The system is still a prototype and not all functionality is fully in place yet. Among the benefits of such a system are a) a fast and systematic reaction in case of pollution events, b) Doramapimod price a spatially and temporal optimized monitoring, c) accelerated alerting and pentoxifylline communication with subsequent reduced heath risk for the local population and tourists, d) an improved awareness, knowledge and transparency about water quality issues, and e) the support of beach profile development and evaluation according to Directive (2006/7/EC). The development of the system or of parts is pushed forward by IMGW PIB (Institute of Meteorology and Water Management-National Research Institute). The web portal www.baltyk.pogodynka.pl

can serve as an example. The system is still not able to serve as a reliable early-warning system for pollution entering with the river. The permanently recording sensor for particulate matter in the river does not sufficiently indicate microbial pollution. The online simulation tool in the Internet information system is a simplified version of the described GETM flow and GITM particle tracking model. It allows end-users to carry out simple but flexible and fast simulations e.g. after accidental release of microorganisms in the coastal area or after the observation of high concentrations at beaches. In a first step the end-user enters the wind situation (direction and speed). The information system contains pre-simulated and stored, steady state flow simulations for altogether 16 wind situations (combinations of direction and speed). The system uses the one that reflects the users demand best.

, 2011). This work was supported by the Max Planck Society, the German Federal Ministry of Education and Research (Microbial Interactions in Marine Systems project (MIMAS), grant number 03F0480D) and the Micro B3 project. The Micro B3 project is funded from the

European Union’s Seventh Framework Programme (Joint Call OCEAN.2011‐2: Marine microbial diversity — new insights Dasatinib price into marine ecosystems functioning and its biotechnological potential) under the grant agreement no 287589. “
“Prochlorococcus is a marine unicellular cyanobacterium that numerically dominates the phytoplankton in the oligotrophic open oceans between 40°N and 40°S ( Partensky et al., 1999). At the northern tip of the Gulf of Aqaba (Station A, 29°28′N 34°55′E) Prochlorococcus reaches up to 2 × 105 cells per mL during the summer at the height of stratification ( Lindell and Post, 1995). Therefore, this sampling site was chosen in order to search for selleck Prochlorococcus-specific transcripts. A number

of distinct Prochlorococcus ecotypes are found in the oceans ( Scanlan et al., 2009) and are divided into two groups according to their ability to adapt to low light (LL) or high light (HL) conditions ( Moore et al. 1995). Despite Prochlorococcus’ compact genomes and relatively low number of transcriptional protein regulators ( Scanlan et al., 2009), this organism is capable of adapting to environmental perturbations suggesting Thiamet G a crucial role of other types of regulators. Indeed, a relatively high number of non-coding (nc)RNAs and antisense RNA have been found in Prochlorococcus by computational prediction, microarrays and high throughput sequencing ( Axmann et al., 2005, Steglich et al., 2008, Richter et al., 2010, Waldbauer et al., 2012 and Voigt et al., 2014). Most studies so far have focused on ncRNAs of the HL-adapted Prochlorococcus strain MED4 in laboratory cultures; here we aimed to identify novel ncRNAs expressed under natural conditions as well as those specific to LL-adapted Prochlorococcus ecotypes. Sampling for metatranscriptome analyses was performed at Station A

in the Gulf of Aqaba (29°28′N 34°55′E) on 14 September 2010 from 60 m (casts 2 and 3 at 9:30 am and 10:10 am respectively), the deep chlorophyll maximum (DCM, ~ 100 m; casts 4 and 5 at 11:30 am and 12:35 pm respectively) and 130 m (casts 6 and 7 at 3:00 pm and 4:20 pm respectively). The water column was stratified at all collection depths as indicated by a constant increase in density of 1 kg m− 3 from the surface down to 150 m (Fig. 1A). Temperature decreased from 26.5 °C at the surface to 22.5 °C at 150 m depth (Fig. 1A). At around 400 m pronounced pycno‐ and thermoclines were visible (Fig. 1A). Chlorophyll a concentration (an indicator of phytoplankton biomass) was 0.25 μg L− 1 at the surface and peaked at around 100 m (DCM) with 0.64 μg L− 1 ( Fig. 1A). At the other two sampling depths chlorophyll a concentrations of 0.37 μg L− 1 (60 m) and of 0.

05 (corrected for multiple comparisons). The weighted sum of parameters estimated in the individual analyses consisted of “contrast” images, which were used for group analyses ( Friston et al., 1999). So that inferences could be made at a population level, individual data were summarized and incorporated into a random-effect model ( Friston et al., 1999). SPMt and SPMZ for contrast images were created as described above. Significant signal changes for each contrast were assessed by means of t-statistics on a voxel-by-voxel basis ( Friston et al., 1999). The threshold for the SPMZ of group analyses was set

at P<0.05 (corrected for multiple comparisons). Anatomical localizations of significant voxels within clusters were achieved using Talairach Demon software ( Lancaster et al., 2000). Anatomical MRI was performed using a Philips Achieva 3.0TX (Royal Philips Electronics, Eindhoven, the Netherlands) GDC-0068 in vivo to permit registration of magnetic source locations with their respective anatomical locations. Before MRI, five adhesive markers (Medtronic Surgical Navigation Technologies, Natural Product Library chemical structure Broomfield, CO) were attached to the skin of the participant’s head (first and second markers located at 10 mm in front of the left tragus and right tragus, third at 35 mm above the nasion, and fourth and fifth at 40 mm to the right and left

of the third marker). MEG data were superimposed on MR images using

information obtained from these markers and the MEG localization coils. Data are presented as mean±SD unless otherwise stated. All P values were two-tailed, and values less than 0.05 were considered statistically significant. Statistical analyses were performed using IBM SPSS 20.0 software (IBM, Armonk, NY). We wish to thank Manryoukai Imaging Clinic for performing MRI and Forte Science Communications for editorial help with the manuscript. “
“The vestibular system has traditionally been thought of as a balance apparatus that is related to brain disorders only when co-morbid symptoms include balance compromise, such as in Meniere′s disease and Parkinson′s disease. However, accumulating research suggests an association between vestibular function and psychiatric disorders, even when balance is apparently unaffected. Acyl CoA dehydrogenase Recent research has described the vestibular system as a potential window for exploring brain function beyond that of maintenance of balance, and into areas of perception, cognition, and consciousness (Lopez and Blanke, 2011). Existing research describes clear links between symptoms of anxiety and depression and the vestibular apparatus, and there is some preliminary evidence suggesting a link between the vestibular system and symptoms of psychosis and mania. Aspects of cognition, particularly spatial memory and spatial perception, have also been linked to vestibular function.

To evaluate such a prospect, it is informative to consider the neural bases of inhibitory control in other domains beside the motoric domain. Research suggests that right lateral prefrontal regions also play a prominent role when the retrieval of

information from episodic memory must be inhibited. Anderson and colleagues [17] devised a mental analog of the Go/No-Go task, called the Think/No-Think task, in which individuals learn associations between cue-target item pairs. In the critical phase of the task, participants are shown just the cue. For some cues, participants are signaled to remember the associated item. For other cues, participants are signaled to www.selleckchem.com/products/dabrafenib-gsk2118436.html inhibit thinking about the associated item. Behavioral results indicate that the more chances an individual has to remember an item associated with a cue, the better the recall compared to items in which no retrieval from memory has been prompted. Likewise, the more chances an individual has to inhibit retrieval, the poorer the recall compared to items in which no retrieval from memory has been prompted. Hence, the Think/No-Think

task focuses on inhibition of retrieval from memory, akin to the inhibition of a motor response in the Go/No-Go task. Neuroimaging work has shown that the right lateral prefrontal cortex plays a prominent role in inhibiting memory retrieval by down-regulating activity in the hippocampus [18••] as well as sensory regions (e.g., ventral visual processing areas) that support the originally encoded memory (e.g., of a visual scene) 18•• and 19•. The region so identified, right middle frontal gyrus (rMFG), is a bit more superior to that identified Trametinib mouse in motor inhibition. Similarly, when individuals are directed to encode and then forget certain items or lists in the directed forgetting paradigm, right hemisphere

regions, including lateral prefrontal cortex, become more active (e.g., 20 and 21 and see [22] for review of neural mechanisms involving inhibitory effects on memory including those at encoding). The rMFG is implicated as being especially important based on a number of findings. For example, activation of rMFG predicts the degree to which individuals are successful at inhibition of memory retrieval, and those individuals with a more negative correlation between activation of the rMFG and the hippocampus are better at suppressing Bumetanide memory retrieval [18••]. In addition, although young adults with ADHD are no worse at retrieving memories (i.e., have equivalent performance on Think trials), they have a specific deficit in the inhibition of memory (i.e., have a poorer ability to inhibit retrieval on No Think trials) (Figure 3a). Importantly, the only brain region in which they show reduced activity as measured by fMRI compared to controls on No-Think trials is the rMFG [23] (Figure 3b), implicating this region as playing a central role in inhibiting memory retrieval.

1). A 1/20 sloping beach was constructed from concrete. This slope angle is consistent with previous studies where mild slopes have varied from 1/15 (Li and Raichlen, 2003), to 1/20 (Synolakis, 1987) to 1/24 (Klettner, 2010), to 1/35 (Grilli et al., 1994). The water height was measured using 12 resistance probes

distributed along the length of the flume and a probe monitor (manufactured JAK inhibitor in-house by HR Wallingford). The resistance probes were calibrated prior to each series of experiments due to their sensitivity to the conductivity of water. The sampling frequency was 50 Hz (so a temporal resolution of ±0.02 s), and the accuracy of wave elevation measurements was ±0.005 m. Runup was measured directly using a horizontal tape measure along the flume wall and recording the maximum penetration point of the first swash (accuracy ±0.01 m), along the centre line of the channel i.e., mid-distance between the RGFP966 flume walls, in order to avoid edge effects. For the runup tests presented in this paper, the surface elevation nearest the wave generator was used to determine the wave parameters (see Fig. 1), and the ratios of a/ha/h ranged between 0.02 and 0.18, for both elevated and N-waves. The advantage of the adopted pneumatic generator is that long and leading

depressed waves could be generated and were stable over the flume length. The wavelengths reproduced were much longer than the ones previously studied. The disadvantage was that some wave reflection occurred at the beach when elevated and leading elevated N-waves were created, due to the relative length of the waves. The measurement of runup is important for comparing the characteristics Methisazone of the present waves with existing studies. Runup was estimated from the measured runup length RlRl and converted to a vertical distance using: equation(7) R=Rltanβ.R=Rltanβ.Wave period and wavelength were retrieved from the wave elevation time series. In many cases the second half of the positive part of the wave does not strictly correspond to the direct signal, due to the reflected waves travelling back from the beach. The period T   and wavelength L   are calculated

using the first half of the positive wave, assuming symmetry (a schematic graph within Fig. 1 illustrates the method used to estimate the wave period): equation(8) T=2(tηmax-t0),T=2tηmax-t0, equation(9) L=cpexpT.L=cpexpT.In (8), tηmaxtηmax is the time of occurrence of the wave peak, and t0t0 corresponds to the time when the value of the wave elevation is 1% of the maximum wave height (tηmax>t0tηmax>t0 and we set T1=tηmax-t0T1=tηmax-t0) prior to tηmaxtηmax. In (9), cpexpcpexp is the wave speed, determined from the experiments, by calculating the temporal correlation between adjacent wave probes. For N-waves, the trough does not trigger any reflections from the slope, so the parameters corresponding to the negative part of the wave are calculated on the full negative profile.

Conidial concentration was measured with a hemacytometer and adjusted to 2 × 106 conidia mL− 1 for inoculation. Five seedlings per pot (6 cm diameter) NVP-BKM120 were inoculated at the 4-leaf stage

with 15 mL of conidial suspension (2 × 106 conidia mL− 1) using an airbrush sprayer. After inoculation, the seedlings were placed in sealed plastic bags at room temperature to maintain 70%–90% humidity. Rice seedlings were returned to the greenhouse 24 h after incubation. Disease reactions were recorded 7 days post-inoculation using a 0–5-scale rating system (0–1: resistance; 2–5: susceptible) [32]. Each experiment was repeated three times. AVR-Pita1, in the plasmid vector PCB980, was co-introduced with plasmid PCB1003, containing the selection marker HyB resistance, using PEG-mediated transformation. AVR-Pita1

was introduced into four isolates, TM2, ZN19, B2 and B8. A total of 100 putative recombinant fungal colonies were grown on HyB-containing media. The 100 colonies were isolated and stored for subsequent experiments. As a KU-60019 ic50 control, four isolates were transformed with the selectable marker (PCB1003) alone to determine whether the transformation and protoplast process had any effect on virulence. To confirm that AVR-Pita1 had been introduced into the protoplasts regenerated from isolates TM2, ZN19, B2 and B8, PCR using the primer pair YT4/YT5 was used to amplify the AVR-Pita1 coding region from 100 putative transformants. Plasmid PCB980 containing AVR-Pita1 was used as a positive control and genomic DNA from the non-AVR-Pita1-containing transformants (without PCB980) was used as a negative control. A total

of 29 transformants were identified by PCR screening as carrying newly introduced AVR-Pita1 ( Fig. 1). A PCR product of 675 bp was repeatedly amplified in both the 29 putative transformants and the positive control ( Fig. 1). No product was amplified from the negative control. These results indicate that AVR-Pita1 was successfully introduced into the virulent U.S. isolates. To verify the identity of AVR-Pita1 in the transformants, amplified PCR products were sequenced and verified with the sequence of AVR-Pita1 amplified from PCB980. Both sequences enough of AVR-Pita1 were identical, suggesting that the PCR product amplified was from the AVR-Pita1 coding region originally cloned in PCB980. To determine the copy number of transformants, the AVR-Pita1 coding region was used as a probe for Southern blot analysis. Multiple hybridization bands of different sizes were found in most transformants. These results suggested that the copy numbers of transformants varied from one copy in recombinant R12 to 15 copies in recombinant R1 ( Fig. 2). Partial incorporation of the transgene occurred in some cases, given that some bands with size around 1 kb (< 1.5 kb AVR-Pita1 promoter plus coding region) appeared on the membrane ( Fig. 2).

Appreciation

is also extended to Dr. Stephanie from Colorado University at Boulder, for her help in refining the language usage. “
“Eleven years after Crutzen (2002) suggested the term Anthropocene as a new epoch of geological time (Zalasiewicz et www.selleckchem.com/products/CP-690550.html al., 2011a), the magnitude and timing of human-induced change on climate and environment have been widely debated, culminating in the establishment of this new journal. Debate has centred around whether to use the industrial revolution as the start of the Anthropocene as suggested by Crutzen, or to include earlier anthropogenic effects on landscape, the environment (Ellis et al., 2013), and possibly climate (Ruddiman, 2003 and Ruddiman, 2013), thus backdating it to the Neolithic revolution and possibly beyond Pleistocene megafauna extinctions

around 50,000 years ago (Koch and Barnosky, 2006). Here, we appeal for leaving the beginning of the Anthropocene at around 1780 AD; this time marks the beginning of immense rises in human population and carbon emissions as well as atmospheric CO2 levels, the so-called “great acceleration”. This also anchors the Anthropocene on the first measurements of atmospheric CO2, confirming the maximum level of around 280 ppm recognized from ice cores to be typical for the centuries preceding the Anthropocene (Lüthi et al., 2008). The cause of the great acceleration was the RG7420 increase in burning of fossil fuels: this did not begin in the 18th century, indeed coal was used 800 years earlier in China and already during

Roman times in Britain ( Hartwell, 1962 and Dearne and Branigan, 1996), but the effects on atmospheric CO2 are thought to have been less than 4 ppm until 1850 ( Stocker et al., 2010). The Anthropocene marks the displacement of agriculture as the world’s leading industry ( Steffen et al., 2011). However, the beginning of the Anthropocene is more controversial than its existence, and if we consider anthropogenic effects on the environment rather than on climate, there is abundant evidence for earlier events linked to human activities, including land use changes associated with the spread of agriculture, Sodium butyrate controlled fire, deforestation, changes in species distributions, and extinctions (Smith and Zeder, 2013). The further one goes back in time, the more tenuous the links to human activities become, and the more uncertain it is that they caused any lasting effect. The proposition of the Anthropocene as a geological epoch raises the question of what defines an epoch. To some extent this is a thought experiment applied to a time in the far future – the boundary needs to be recognizable in the geological record millions of years in the future, just as past boundaries are recognized.