The authors thank the Brazilian Funding Agencies FAPESP and CNPq

The authors thank the Brazilian Funding Agencies FAPESP and CNPq for their financial support. “
“Cafestol (1) and kahweol (2) (Fig. 1) are two examples of naturally-occurring furan diterpenes in the lipid fraction of coffee (Bengis and Anderson, 1932, Djerassi et al., 1953, Haworth et al., 1954, Dias et al., 2010, Haworth and Johnstone, 1956 and Lam

et al., 1982). Of the two most significant species in the AZD2281 purchase coffee trade, Coffea arabica L. (Arabica) contains about 0.6% of cafestol (1) and 0.3% of kahweol (2) while Coffea canephora Pierre (Robusta) contains mostly cafestol (1) (0.2%) and 16-O-methylcafestol (0.15%), not present in Arabica coffee ( De Roos et al., 1997, Nackunstz and Maier, 1987, Pettitt, 1987 and Ratnayake et al., 1993). Coffee furan diterpenes are mainly present in the esterified form, with 26 different fatty acids, and only small amounts are in the free form ( Fig. 1) ( Kurzrock & Speer, 2001). The amount of diterpenes present in the coffee brew

depends on the way coffee is prepared. The highest content of diterpenes was found in boiled, unfiltered coffee brews, while in drip-filtered coffee brews they are Bortezomib nmr negligible next (Martín, Pablos, González, Valdenebro, & León-Camacho, 2001). Cafestol and kahweol have been described to be both desirable

and undesirable in human health. They are anticarcinogenic (Butt and Sultan, 2011, Cavin et al., 2002, Hatzold, 2012, IARC, 1991, Kim et al., 2009, Lam et al., 1982, Lee et al., 2007 and Tao et al., 2008), antioxidant (IARC, 1991 and Lee and Jeong, 2007) and anti-inflammatory (Bertholet, 1987) and showed hepatoprotective effects (Lee et al., 2007). On the other hand, a hypercholesterolaemic action has been reported, attributed mainly to cafestol (Arnesen et al., 1984, Butt and Sultan, 2011, Hatzold, 2012, Urgert et al., 1995 and Weusten-Van Der Wouw et al., 1994), and they are also responsible for increasing low-density lipoprotein (Urgert & Katan, 1997). Green coffee oil is obtained by mechanical cold pressing or extraction with solvents, such as hexane, but these traditional methods are labour intensive and time-consuming, and sometimes require large quantities of solvents (Araujo & Sandi, 2006). Other procedures involve supercritical fluid extraction method (SFE) (Araujo and Sandi, 2006 and De Azevedo et al., 2008) and high-speed countercurrent chromatography (HSCCC) (Scharnhop & Winterhalter, 2009).

Then, 30 mL of the solution was aseptically transferred

u

Then, 30 mL of the solution was aseptically transferred

using a serologic pipette to sterile petri dishes (inner diameter 15.6 cm; Sarstedt Ltd., Leicester, UK). The cast solutions were air dried at 37 °C for 15 h in a ventilated incubator (Sanyo Ltd., Japan) in order to obtain films that could be easily peeled off and had acceptable mechanical properties (absence of brittleness and adequate flexibility/extensibility). After drying, the probiotic edible films were peeled intact from the petri dishes and conditioned at room (25 ± 1 °C) selleck chemicals llc or chilled temperature (4 ± 1 °C) under controlled relative humidity conditions (54% RH) in desiccators containing saturated magnesium nitrate solution. One mL of the probiotic film forming solution was suspended in sterile PBS and vortexed for 30 s to ensure adequate mixing using the method described by Lopéz de Lacey et al. (2012) with minor modifications.

More specifically, individual 1 g film samples containing L. rhamnosus GG were transferred to 9 mL of sterile PBS and left to hydrate and dissolve under constant agitation in an orbital incubator at 37 °C for 1 h. The complete dissolution of the edible films had been previously been tested using edible films without probiotics and no residual insoluble material could be identified. In both cases, the resulting solutions were subjected to serial dilutions using phosphate buffer saline. Each dilution was pour plated on a MRS agar

(MRS Agar, Oxoid Ltd., Basingstoke, UK) and the selleck plates were stored at 37 °C for 72 h under anaerobic conditions to allow colonies to grow. Enumeration of the bacteria on agar plates was performed in triplicates by colony counting ( Champagne, Ross, Saarela, Hansen, & Charalampopoulos, 2011) and the total counts of the viable bacteria were expressed as log colony forming units per gram (log CFU/g, CFU/g = CFU/plate × dilution factor). The survival rate of the bacteria throughout the film forming solution drying process was calculated according to the following equation: equation(1) %viability=100×NN0where N0, N represent the number of viable bacteria prior and after the implemented drying process ( Behboudi-Jobbehdar et al., 2013). L. rhamnosus GG inactivation upon storage data was expressed as Oxalosuccinic acid the value of the relative viability fraction N/N0. The viability data were fitted to a first order reaction kinetics model as described by the formula: equation(2) Nt/N0=1-kTtNt/N0=1-kTtwhere N0 represents the initial number of the viable bacteria and Nt the number of viable bacteria after a specific time of storage (in CFU/g), t is the storage time (in day), and kT is the inactivation rate constant at T temperature (day−1). A digital micrometre with a sensitivity of 0.001 mm was used for the measurement of the thickness of the probiotic edible films. Eight measurements were taken from different parts of the films to ensure results consistency.

Also, the spectrophotometric

analyses were performed in t

Also, the spectrophotometric

analyses were performed in triplicate for each wine. The free radical scavenging activity of the wine samples was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenger method measured at 518 nm (Brand-Williams, Cuvelier, & Berset, 1995) and ABTS 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) according to Re et al. (1999), measured at 754 nm. Lipid peroxidation Selleckchem ABT-263 inhibition was assayed using the TBARS method, as described by Chen and Tappel (1996). Results were expressed as Trolox equivalents (mm TEAC). The analyses were carried out in triplicate. Analysis of variance (ANOVA), the Tukey HSD Test and PCA were carried out using Statistica 7 (2006) (StatSotft Inc., Tulsa, OK) and

p < 0.05 values were considered statistically significant. The GSI-IX clinical trial linear regression, the square of the correlation coefficient of the regression line, and the limits of detection and quantitation obtained from the calibration data for catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, PA B1 and PA B2 standards are shown in Table 1. The % RSD obtained experimentally with 12 analyses of the wine sample were as follows: for free flavan-3-ols: catechin, 3.80%; epicatechin, 3.78%; gallocatechin, 4.04%; epigallocatechin, 2.87%; PA B1, 3.86%; and PA B2, 3.56%; for proanthocyanidins, terminal units: catechin, 4.71%; epicatechin, 4.07%; gallocatechin, 4.03%; epigallocatechin, 3.06%; and epicatechin gallate, 4.57%; and extension units: catechin, 6.75%; epicatechin, 3.17%; epigallocatechin, 1.87%; and epicatechin gallate 6.26%. All results were considered acceptable for research purposes. The flavan-3-ol monomers catechin (C), epicatechin (EC), gallocatechin (GC) and epigallocatechin (EGC) and PA dimers B1

and B2 were identified and quantified in wine samples of Cabernet Franc, Merlot, Sangiovese and Syrah, from 2006 and 2007 vintages, from São Joaquim – SC, Brazil (Fig. 1, Table 2). The main flavan-3-ol monomers found were catechin and epicatechin. These results are in agreement with those in the literature, since these P-type ATPase are the main monomers in the skin and seeds of grapes (Chira et al., 2009, Mattivi et al., 2009 and Prieur et al., 1994) and, consequently, in wine. Catechin was the main monomer in the wine samples evaluated, with the highest concentrations observed in all samples, representing, on the average, 60% of the total monomers, as also observed in other studies (Monagas, Gómez-Cordovés, Bartolomé, Laureano, & Ricardo da Silva, 2003). The highest concentrations of catechin were observed in Merlot 2007 and Syrah 2006 samples. Epicatechin represented approximately 25% of the monomers quantified in the samples, with concentrations ranging from 4 to 16 mg L−1, Merlot and Syrah being the varieties showing the highest concentrations.

5, personally monitored exposure showing the strongest associatio

5, personally monitored exposure showing the strongest associations followed by NVP-BEZ235 in vivo indoor PM exposure, and then outdoor and central-site measurements (Delfino et al., 2004). A 3-year panel study on children with asthma and adults with or without chronic obstructive pulmonary disease found inverse associations between lung function and exposure

to PM2.5 in both adults and children with lung disease and most consistently with respect to indoor exposures (Trenga et al., 2006). Most studies of healthy individuals have reported no associations between indoor PM2.5 and lung function (Ebelt et al., 2005, Jansen et al., 2005 and Yeatts et al., 2012). Two studies including both smokers and subjects who were exposed to environmental tobacco smoke, but otherwise healthy, have shown associations between lung function or symptoms with

indoor concentrations of PM2.5 in a panel study of elderly especially during winter (Simoni et al., 2003) and in an indoor air filtration crossover study with young adults (Weichenthal et al., 2013). By contrast, our investigation encompassed only non-smokers without asthma, living in non-smoking homes and lung function was not associated with PM2.5 only with PNC levels. Possibly specific effects of high outdoor PNC levels from traffic have been found in adults with asthma, showing decreased lung function after short-term exposure in traffic-dense environments INCB024360 cell line (McCreanor et al., 2007). An exposure contrast in PNC (9000–66,500 particles/cm3) else for 5 h while exercising intermittently at five different locations including two traffic sites, an urban background location, an underground train station and a farm in the Netherlands was associated with decreased lung function in young healthy subjects (Strak et al., 2012). However, in healthy young adults no effect on lung function was observed during 24 h of exposure to air from a busy street in Copenhagen, Denmark, with PNC of 6000–15,000 particles/cm3

(Brauner et al., 2009). Similarly, a 2-hour exposure to high PNC in a road tunnel (1.3 × 105 particles/mL) or concentrated ambient UFP (2.1 × 105 particles/cm3) were not associated with altered lung function in young and healthy subjects (Larsson et al., 2007 and Samet et al., 2009). Many studies on the associations between air pollution-mediated systemic inflammation and cardiovascular diseases have assessed CRP and leukocyte counts as markers of inflammation (Delfino et al., 2005). We found a significant positive association between indoor exposure to PM2.5 and elevated levels of CRP. We also found positive associations between outdoor particle levels and CRP, but they were not statistically significant. A 7-day intervention study with air filtration in the homes of a wood smoke impacted area found an association between the indoor concentration of PM2.5 and CRP (Allen et al.

Several explanations have been proposed for this pattern of resul

Several explanations have been proposed for this pattern of results. First, children’s failure at the last task suggests that keeping the sets visible may have a negative impact on their performance. When sets are visible, children may be drawn to rely on perception, which is approximate, and thus to generalize number words beyond exact numerical quantities. However, this explanation seems MEK inhibitor unlikely, because (1) Condry and Spelke’s (2008) visible single-set task induced major changes in numerosity (doubling and halving), easily detectable by children, and (2) children failed at Sarnecka and Gelman’s (2004) one-to-one comparison task, where the conditions of presentation

highlighted any difference across sets. Second, it is possible that tasks involving two sets are simply overwhelming for children, single-set tasks thus being a better indicator of children’s this website semantic competence (Sarnecka & Gelman, 2004). However, Condry and Spelke (2008) showed that children sometimes succeed in two-set tasks, since participants solved the task with high accuracy when no transformation was applied to the sets, and they also showed that participants sometimes failed in single-set tasks. Third, counter to the previous explanation, Brooks et al. (2012) argued that children succeeded at Sarnecka and Gelman’s (2004) single-set

transformation task without extensive knowledge of the semantics of the number words. According to their argument, to succeed at the task children only need to know that a change in quantity is necessary to warrant a change of number word: therefore, children know to conserve

the initial label after a shaking event. For addition and subtraction transformations, however, they find the right answer only by applying pragmatic inferences: If a child is given a choice between a label he/she heard earlier 4��8C in the trial and a new label, Brooks et al. argue, given the assumption that the adult asking the question is knowledgeable, the child would infer that the new label provided is relevant. Pragmatic inferences, in contrast, provide no ground to find the correct answer in Condry and Spelke’s (2008) two-sets task. To support this view, Brooks et al. adapted Condry and Spelke’s (2008) two-set task and Sarnecka and Gelman’s (2004) single-set transformation task using novel words and objects, and obtained the same pattern of success and failure across these two tasks, where children were asked to choose between two labels (as in Sarnecka and Gelman’s single-set task) or between two objects (as in Condry and Spelke’s two-sets task). This last explanation holds promise to explain the whole set of results, with one adjustment: Given the contrast between children’s reasoning about identity and substitution events in Experiment 4, children may not think that a change in number words requires a change in quantity but rather a change of set identity.


“Leaf area as photosynthetically active area is one of


“Leaf area as photosynthetically active area is one of

the main drivers for tree growth and thus an important tree characteristic for tree growth studies. For silvicultural purposes trees have to be considered as parts of stands, and individual tree growth has to be investigated in relation to stand structure. Thus, O’Hara (1988) PCI-32765 concentration used the area for individual trees as a measure of site occupancy. Leaf area in relation to stand parameters, e.g., ground area potentially available (APA), which could be named as individual tree leaf area index, but also leaf area in relation to stemwood increment which is described as growth efficiency (Waring, 1983) are important research issues. However, leaf area is hard to determine precisely and non-destructively. For leaf area index determination of stands various optical instruments like LAI-2000 (Li-Cor) or SunScan (Delta-T) are available. But these instruments are limited by the complexity of the canopy structure and improvement in accuracy is still needed (Moser et al., 1995, Chen et al., 1997, Pokorny and Marek, 2000 and Pokorny et al., 2004). Another

way to determine stand leaf area index is to use the individual tree leaf area. Hence, different approaches to estimate individual tree leaf area in an indirect way were and are investigated. Such investigations aim at strong relations of leaf area to other tree characteristics. Based on the pipe model of Shinozaki et al. (1964), BEZ235 in vitro which supposed that a given leaf area is supplied ifenprodil with water from a respective quantity of conducting pipes, mainly sapwood area (e.g., Waring et al., 1982, Bancalari et al., 1987 and Meadows and Hodges, 2002), early sapwood area (Eckmüllner and Sterba, 2000), and diameter at breast height (e.g., Gholz et al., 1979 and Baldwin, 1989) are used as estimators for leaf area or leaf biomass. A few studies deal with estimating leaf area with allometric functions based on different other

tree characteristics (e.g., Pereira et al., 1997 and Kenefic and Seymore, 1999). The majority of studies dealing with indirect leaf area estimation describe sapwood area as the most accurate estimator for leaf area (e.g., Long et al., 1981, O’Hara and Valappil, 1995 and Meadows and Hodges, 2002). But to get continuously information about leaf area and related characteristics, e.g., growth efficiency, and their development over time, the determination via sapwood area is not feasible, because from the same trees cores cannot be taken every 5 or 10 years over a long term. Additionally, it is well known that the relationship between leaf area and sapwood area, even within species, is not constant. Differences could be shown between sites, crown classes, stand density, and age (Long et al.

25 U of Taq DNA polymerase (Fermentas), and 0 2 mmol/L of each de

25 U of Taq DNA polymerase (Fermentas), and 0.2 mmol/L of each deoxyribonucleoside

triphosphate (Biotools, Madrid, Spain). Positive and negative controls were included in each batch of samples analyzed. Positive controls consisted Baf-A1 supplier of DNA extracted from Porphyromonas gingivalis (ATCC 33277), Methanobrevibacter arboriphilus (DSMZ 744), and Candida albicans (ATCC 10231). Negative controls consisted of sterile ultrapure water instead of sample. All reactions were run in triplicate. PCR amplifications were performed in a DNA thermocycler (Mastercycler personal; Eppendorff, Hamburg, Germany). Cycling conditions were as follows. For bacteria, it included initial denaturation step at 95°C for 2 minutes, followed by 36 cycles at 95°C/30 seconds, 60°C/1 minute, and 72°C/1 minute, and final extension at 72°C/10 minutes. For archaea, it included initial denaturation at 94°C/2 minutes, 36 cycles at 94°C/30 seconds, 58°C/30 seconds, and 72°C/1 minute, and final extension at 72°C/10 minutes. For fungi, it included initial denaturation step at 95°C/30 seconds, followed by 40 cycles at 95°C/30 seconds, 55°C/1 minute, 72°C/2 minutes, and a final step at 72°C/10 minutes. PCR products were subjected to electrophoresis in a 1.5% agarose gel–Tris-borate-EDTA

buffer. The gel was stained with GelRed (Biotium, Hayward, CA) and visualized under ultraviolet selleck chemical illumination. The presence of amplicons of the expected size for each primer pair was considered as positive result. A 100 base pair DNA ladder (Biotools) was used as a parameter for amplicon size. For bacterial identification in the checkerboard assay, a practically

full-length 16S rRNA gene fragment was amplified by using universal bacterial primers 8f and 1492r, with the forward primer labeled at the 5′ end with digoxigenin. PCR amplifications were performed as see more described above for bacteria. The reverse-capture checkerboard assay was conducted to determine the presence and levels of 28 bacterial taxa as described previously 20, 24 and 25. Probes were based on 16S rRNA gene sequences of the target bacteria and were described and validated elsewhere 20, 24, 26 and 27. Prevalence of the target taxa was recorded as the percentage of cases examined. A semiquantitative analysis of the checkerboard findings was conducted as follows. The obtained chemiluminescent signals were evaluated by using ImageJ (W. Rasband, http://rsb.info.nih.gov/ij/) and converted into counts by comparison with standards at known concentrations run on each membrane.

To screen for the best-performing siRNA of each group, we employe

To screen for the best-performing siRNA of each group, we employed a dual-luciferase assay-based system. The respective target sequences were individually inserted into the 3′ UTR of a plasmid-located Renilla luciferase

gene. The DNA polymerase, pTP, IVa2, hexon, and protease siRNAs, together with GSK1210151A concentration the respective reporter vectors, were used to co-transfect HEK293 cells. Knockdown of Renilla luciferase expression in relation to the expression of a firefly luciferase gene located on the same plasmid was determined in dual-luciferase assays. The silencing capacity of the E1A siRNAs was assessed in A549 cells because promoter activities of the reporter vectors turned out to be altered upon silencing of the endogenous E1A gene present in HEK293 cells ( Graham et al., 1977) (data not shown). For all target mRNAs,

we identified siRNAs enabling a knockdown of ⩾78% at a concentration of 30 nM ( Fig. 1). The best-performing siRNAs of each group, i.e., pTP-si8, Pol-si2, Hex-si2, E1A-si3, Iva2-si2, and Prot-si1, were selected for further characterization. The dual-luciferase assay-based screening system was employed to select the best-performing siRNAs of each group. Next, we investigated whether the selected siRNAs were able to knockdown gene expression during an adenovirus infection of A549 cells. Cells were transfected with the siRNAs at a concentration of 10 nM, and then infected with Ad5 at an MOI of 0.01 TCID50/cell. Target mRNA levels were determined SCH772984 nmr by RT-qPCR, using primers specific for the individual mRNAs (Fig. 2A).

The highest silencing rates (93–97%) were observed for the E1A-, DNA polymerase-, pTP-, and IVa2-targeting Sulfite dehydrogenase siRNAs. Silencing of the hexon and protease genes was less pronounced (79–87%). Except for the difference in residual hexon and pTP mRNA levels, the differences between hexon or protease mRNA levels and those of all other early genes were statistically significant. As the pTP, DNA polymerase, and IVa2 mRNAs share a common 3′ part (Supplementary Fig. 1), and the DNA polymerase target site is also part of the pTP mRNA, IVa2- and DNA polymerase-directed siRNAs were therefore expected to concomitantly silence pTP/DNA polymerase/IVa2 and pTP/DNA polymerase, respectively. Furthermore, siRNA-mediated silencing of early genes was expected indirectly to affect the expression of those middle or late genes for which expression is known to depend on early viral gene products. Thus, we also determined the effect of the E1A-, pTP-, DNA polymerase-, and IVa2-targeting siRNAs on the expression of the other genes. Silencing of E1A resulted in a marked reduction in the expression of all other genes (Fig. 2B). This can be attributed to the central role of E1A in activating the expression of downstream genes. Silencing of the E1A gene actually resulted in a greater reduction in the expression of hexon and protease than did direct silencing of these genes by the hexon and protease siRNAs (compare Fig. 2A and B).

In the survey, students were shown an identical series of photos

In the survey, students were shown an identical series of photos of river segments and asked to rate each river segment on a numerical scale in terms of being natural, esthetically pleasing, dangerous,

and needing ABT-888 nmr improvement. With the exception of the U.S. state of Oregon, and the countries of Germany and Sweden, students consistently rated river segments containing instream wood negatively, viewing these river segments as unnatural, dangerous, and in need of rehabilitation (Chin et al., 2008). This completely contradicts the manner in which river scientists view instream wood, and ignores the logical assumption that, since a much greater proportion of the world was forested historically, most river segments in forested environments would naturally contain a great deal of instream wood (Montgomery et al., 2003). The students’ negative perception of instream wood at least partly reflects the fact that most of them are used to seeing rivers with very little instream wood, even in forested environments, because of historical and continuing wood removal. Wood-poor rivers now seem

normal and natural Selleck Ion Channel Ligand Library to most people. Those of us who work in rivers and are familiar with the scientific literature on instream wood, as well as the idea of dramatic historical change in landscapes and ecosystems, can metaphorically step back and shake our heads at the students’ misperceptions, but identifying our own unexamined and misleading Urease perceptions is much more challenging. The default assumption of greater human manipulation of the landscape appears

to apply broadly to temperate and tropical zones, whether arid, semiarid or humid. Archeologists have developed convincing evidence that the seeming wilderness of the pre-Columbian Amazon basin hosted many more people than initially thought, although estimates range enormously from 500,000 to 10 million people (Mann, 2005 and McMichael et al., 2012) and remain controversial. Certainly some of these people intensively managed the surrounding vegetation and soils, as reflected in the persistence of dark-colored, fertile terra preta ( Liang et al., 2006) soils that were created by pre-Columbian Indians from 500 to 2500 years BP. Prehistoric agricultural societies in central Arizona, USA created an extensive network of irrigation canals that resulted in soil salinization that persists today ( Andrews and Bostwick, 2000). Only very limited areas of high latitude (Antarctica, parts of the Arctic) and high altitude appear not to have been manipulated by humans at some point during the past few millennia ( Sanderson et al., 2002 and McCloskey and Spalding, 1989). Faced with the realization that most landscapes have been and continue to be manipulated by humans in ways subtle or obvious, geomorphologists can make at least three important contributions to sustaining critical zone integrity.

The most obvious

The most obvious AG-014699 solubility dmso and indeed that which was first suggested by Crutzen (2002) is the rise in Global temperatures caused by greenhouse gas emissions which have resulted from industrialisation. The Mid Holocene rise in greenhouse gases, particularly CH4 ascribed to

human rice-agriculture by Ruddiman (2003) although apparently supportable on archaeological grounds ( Fuller et al., 2011), is also explainable by enhanced emissions in the southern hemisphere tropics linked to precession-induced modification of seasonal precipitation ( Singarayer et al., 2011). The use of the rise in mean Global temperatures has two major advantages, firstly it is a Global measure and secondly it is recorded in components of the Earth system from ice to lake sediments and even in oceanic sediments through acidification. In both respects it is far preferable Rapamycin to an indirect non-Earth systems parameter such as population growth or some arbitrary date ( Gale and Hoare, 2012) for some phase of the industrial revolution, which was itself diachronous. The second, pragmatic alternative has been to use the radiocarbon baseline set by nuclear weapon emissions at 1950 as a Global Stratigraphic Stage Age (GSSA) and after which even the most remote lakes

show an anthropogenic influence ( Wolfe et al., 2013). However, as shown by the data in this paper this could depart from the date of the most significant terrestrial stratigraphic signals by as much as 5000 years. It would also, if defined as an Epoch boundary, mark the end of the Holocene which is itself partly defined on the rise of human societies and clearly contains significant and in some cases overwhelming human impact on geomorphological

systems. Since these contradictions are not mutually resolvable one area of current consideration is to consider a boundary outside of or above normal geological boundaries. It can be argued that this is both in the spirit, if not the language, Docetaxel in vivo of the original suggestion by Crutzen and is warranted by the fact that this situation is unique in Earth history, indeed in the history of our solar system. It is also non-repeatable in that a shift to human dominance of the Earth System can only happen once. We can also examine the question using the same reasoning that we apply to geological history. If after the end of the Pleistocene, as demarcated by the loss of all ice on the poles (either due to human-induced warming or plate motions), we were to look back at the Late Pleistocene record would we see a litho- and biostratigraphic discontinuity dated to the Mid to Late Holocene? Geomorphology is a fundamental driver of the geological record at all spatial and temporal scales. It should therefore be part of discussions concerning the identification and demarcation of the Holocene (Brown et al., 2013) including sub-division on the basis of stratigraphy in order to create the Anthropocene (Zalasiewicz et al., 2011).