Brains were washed in phosphate-buffered saline (PBS) (with Ca++/Mg++) and meninges were thoroughly peeled off and discarded. White matter was carefully removed. The grey matter was collected in HEPES-buffered MEM containing 10% foetal calf serum (MEM-H 10% FCS), http://www.selleckchem.com/products/SGI-1776.html forced through a 50 ml syringe to produce a slurry, and mixed with an equal volume of MEM-H 10%. Tissue was gently homogenised in a glass Wheaton Dounce tissue grinder (Jencons Scientific Ltd., Leighton Buzzard, UK) (89–127 μm clearance, 15 strokes;

25–76 μm clearance 15 strokes) and sequentially filtered, first through 150 μm nylon mesh, then through 60 μm nylon mesh. Microvessel fragments trapped on the 150 and 60 μm meshes were kept separate and digested at 37 °C for 1 h in medium M199 containing 10% FCS, 223 U/mg collagenase, 211 U/mg trypsin and 2108 U/mg DNase with continuous agitation. Microvessels were washed off the meshes with the enzyme mixture, centrifuged for 5 min at 240g at 4 °C to remove enzyme, then resuspended in MEM-H 10% FCS and centrifuged again; the resulting vessel fractions were kept separate as ‘150s’ and ‘60s’, the latter giving higher TEER. The ‘60s’ were used for all experiments described here. Digested fragments were resuspended in 10% DMSO in foetal calf serum, brought slowly to −80 °C and stored in liquid nitrogen. Six pig brains gave 12 1 ml aliquots of ‘60s’. Capillary fragments

were thawed and resuspended in plating medium consisting of DMEM with 10% BPDS with 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM glutamine, 125 μg/ml heparin, with 4 μg/ml puromycin to kill contaminating

cells, especially pericytes (Perrière et FK866 cost al., 2005). One aliquot was plated into two T75 flasks coated with lab-prepared rat tail collagen (Strom and Michalopoulos, 1982) and 7.5 μg/ml fibronectin, and grown to 70–80% confluence. Cells were detached by brief trypsinisation (500 BAEE units trypsin and 0.47 mM EDTA.4Na in HBSS without Ca2+ or Mg2+), then centrifuged at 360g for 5 min. The pellet of these first passage (P1) cells was resuspended NADPH-cytochrome-c2 reductase in plating medium containing DMEM, 10% BPDS, 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM glutamine and 125 μg/ml heparin. Cells were seeded onto collagen/fibronectin coated Transwell-Clear inserts at a density of 1×105 cells/cm2 or at 1×104 cells/well in 96-well plates for functional studies and grown for 2–3 day until confluent. The medium was changed to serum-free medium supplemented with 550 nM hydrocortisone ( Hoheisel et al., 1998) and the cells were treated with 250 μM pCPT-cAMP and 17.5 μM RO-20-1724 ( Rubin et al., 1991); these supplements helped to improve differentiation of BBB properties, especially tight junction maturation ( Förster et al., 2005). PBECs were used in experiments 24 h after this medium change. The quality of the model in terms of cell growth was assessed according to the time the cultures took to become confluent.

The authors have no conflicts of interest to declare. This work was partially supported by CAPES-Brazil/MES-Cuba (064/09). “
“The authors regret an error in Methods, under “Test article.” The DMSO concentration should read 0.1% and not 0.01%. The authors would like to apologise for any inconvenience caused. “
“Juglone (5-hydroxy-1,4-naphthoquinone) is a phenolic compound with allelopathic properties belonging to the class of naphthoquinones. Its chemical structure is shown in Fig. 1. This quinone is found in roots, leaves, bark and nuts of several species of walnut from the plant family Juglandaceae (Lee and Campbell,

1969). The α-hydrojuglone is the reduced form of juglone and is related to developmental processes and defense mechanisms of the nuts. When exposed to the air, the α-hydrojuglone ROCK inhibitor is

readily oxidized to Venetoclax solubility dmso juglone (Duroux et al., 1998 and Rietveld, 1983). The extract of walnut is widely used in popular medicine as a phytotherapic to treat inflammatory diseases, eczema, acne, herpes, psoriasis, and bacterial, fungal, viral and parasitic diseases (Bell, 1981, Jin, 2010 and Mahoney et al., 2000). On the other hand, juglone has been investigated by the National Toxicology Program (USA) as a potentially toxic natural product (Mahoney et al., 2000). The naphthoquinones can cause a variety of hazardous effects in vivo, including acute cytotoxicity and immunotoxicity (Bolton et al., 2000). The mechanisms by which juglone causes cell toxicity are

complex. This is partly caused by the fact that juglone can assume three structures which are in equilibrium: in addition to the oxidized and fully reduced forms shown in Fig. 1, the partially reduced semiquinone is also usually present. The mechanisms of action of juglone comprise mixed actions which include the reactivity of the electrophilic quinoidal group Reverse transcriptase and the ability to undergo oxidation–reduction cycles with concomitant formation of free radicals (Duroux et al., 1998, O’Brien, 1991 and Rath et al., 1996). Juglone can also interact with nucleophilic biomolecules such as glutathione and thiol groups of proteins which lead to the oxidation of nucleophilic sites. This, in turn, causes inactivation of enzymes or cellular signaling proteins (Klaus et al., 2010). The toxicity of juglone on bacteria is attributed to changes in the plasma membrane (Zhang et al., 1994). In human lymphocytes, 50 μM juglone inhibits cell proliferation by blocking potassium channels. In consequence it induces polarization of the plasma membrane (Varga et al., 1996). The juglone also appears to inhibit enzymes such as protein kinase C (Frew et al., 1995) and cytochrome P450 aromatase in human placental microsomes in a dose-dependent manner (Muto et al., 1987). It also blocks transcription, induces DNA damage, reduces protein levels and induces cell death (Paulsen and Ljungman, 2005).

These findings suggest that exposure to petroleum compounds result in cell and tissue changes that predispose fish to infectious diseases. One of the objectives of this study was to validate flow cytometry leukocyte counts compared to manual leukocyte differentials. Flow cytometry

allows rapid analyses of large numbers of samples to broadly assess immune status. Flow cytometry results corresponded well to manual leukocyte differentials. DiOC5 and DiOC6 stains were used to aid in the separation of thrombocytes, but did not work consistently. Therefore, the actual number of thrombocytes, or the differentiation of thrombocytes from lymphocytes could not be determined during flow cytometry, so some thrombocytes INNO-406 manufacturer were included in the lymphocyte counts and explain the higher lymphocyte numbers when compared to manual differential lymphocyte counts. The alligator gar peripheral blood counts from the Gulf did not demonstrate significant differences compared to unexposed alligator gar by either the manual or flow cytometric methods. Although there is no direct evidence that oil exposure results in an increased occurrence of disease outbreaks, it is well documented that exposure to petroleum compounds impacts fish immunity, which subsequently affects fish health. Toxins can affect fish directly or indirectly. Furthermore, the effects

vary by compound and concentration, and which specific immune response is being examined. Beginning on April 20, 2010, petroleum and dispersant

compounds Pregnenolone associated with the Macondo oil Venetoclax in vivo spill were present in areas of the Gulf of Mexico. We sampled alligator gar, Gulf killifish and sea trout in these locations, and compared them to control fish. A definitive finding was that peripheral blood lymphocyte numbers were significantly reduced in sea trout and Gulf killifish. Lymphopenia is documented to result in decreased disease resistance in vertebrates. Another finding was the number of splenic melano-macrophage aggregates was significantly increased in sea trout and Gulf killifish. The size of splenic melano-macrophage centers was significantly greater in sea trout. Increases in number and size of melano-macrophage aggregates are associated with environmental toxin exposure in fish. A third finding was that liver EROD values from Gulf sea trout were significantly higher than non-exposed controls. Increased EROD levels are associated with PAH exposure in fish. Oil from a large underwater plume had the same signature as the oil from the Macondo well (Camilli et al., 2010, Reddy et al., 2012 and White et al., 2012). A logical corollary to our findings is the fish we sampled in the Gulf of Mexico were exposed to crude oil from the Macondo well, and were immuno-compromised.

Assuming a two state model, the observed mean-residue ellipticity at 222 nm ([Θ]obs222) was converted into α-helix fraction (fH) using the method proposed

by Rohl and Baldwin (1998) and previously described ( Konno et al., 2001). The lipid bilayers were obtained from giant unilamellar vesicles (GUVs), which were positioned onto the chip http://www.selleckchem.com/products/Vincristine-Sulfate.html aperture by application of negative pressure. The GUVs burst as soon as they touch the glass surface of the chip and form a bilayer that spans the aperture (Sondermann et al., 2006). Asolectin (Sigma), a negatively charged mixture of lipids, was used to form artificial membranes. GUVs were formed by electroswelling, using the Nanion Technologies (Munich, Germany) device Vesicle Prep Pro©. 20 μL of 10 mg/mL lipid solution (in chloroform) were deposited onto an indium tin oxide (ITO) coated glass plate and evaporated for 45–60 min. A nitrile ring was placed around the dried lipid film and filled with 350 μL of 250 mM D-Sorbitol dissolved in Milli-Q water. A second ITO coated glass plate was placed on top of the ring. An AC voltage of 3 V peak-to-peak JNK inhibitor molecular weight amplitude at 5 Hz frequency was supplied to the ITO slides over a

period of 2 h at 36 °C (modified from Sondermann et al., 2006). The formed vesicles were kept in plastic vials under refrigeration (4 °C) until use or used immediately. GUVs suspensions were always observed under light microscope prior to use. The experiments were performed with the automated Patch-Clamp device Port-a-Patch (Nanion Technologies – Munich, Germany), using borosilicate glass chips NPC-1 with aperture

diameter of approximately 1 μm. The resistance of the apertures was approximately 1–3 MΩ in 150 mM HCl solution. Current signals were amplified and recorded by an amplifier EPC-10 (Heka Elektronik, Lambrecht, MRIP Germany) and an analogical/digital interface ITC-1600. The system was computer controlled by the PatchControl™ software (Nanion) (Fertig et al., 2002 and Sondermann et al., 2006). During the experiments symmetrical solution of 150 mM HCl with 5 mM Tris was used. After a seal was formed (Rm > 500 mΩ), the peptides diluted with Milli-Q water at a 5 μM concentration were added to the cis side of the chip (top) to observe the single channel activity. The volume of peptide solution was never superior to 10% of the solution at the cis side. Voltage pulses were applied at the trans side of the chip (bottom). Usually, single channel activity started approximately 10 min after adding the peptides, as monitored by a constant Vhold of −100 mV. Single channel conductance of incorporated channels was determined under positive and negative voltage pulses. The experiments were performed at room temperature (∼22 °C). The data was analyzed by PatchMaster and Matlab softwares.

The overlap settings for this assembly were a mismatch cost = 2, an insert cost = 3, a minimum contig length = 200 bp, and a similarity = 0.8. Finally, 36,869 contigs were obtained with an average length of 746 bp ( Fig. 1A). Among these, 11,277 putative transcripts were found exclusively in females, while 5876 contigs were present in just males ( Fig. 1B). All contigs were annotated according to Gene Ontology (GO) terms with the Blast2Go software ( Conesa et al., 2005) and through Blastp against the NCBI non-redundant (nr) protein database built for arthropods, considering a cutoff

E-value of 1E−10. Thus, 13,749 (37%) sequences were annotated, of which just 628 (4.5%) matched Artemia sequences, leaving

13,121 (95.5%) novel putative transcripts for this species ( PF-562271 Fig. 1C). The complete list of annotated contigs is available in Table Tacrolimus price S1. Moreover, GO annotation evidenced a similar distribution of assembled contigs for the female and male datasets ( Fig. 1D). RNA-Seq analysis was conducted by individually mapping the female and male datasets against the 36,869 generated contigs, and the expression level of each transcript was quantified in reads per kilobase of the transcript per million mapped reads (RPKM). The parameters considered included a minimum read length fraction = 0.8, minimum read similarity fraction = 0.9, and unspecific read match limit = 10 in relation to the reference values. Expression values were normalized by totals using state numbers in reads per 1,000,000, and a Kal’s Z-test was conducted using the female group as a reference. A volcano plot was performed to select the differentially expressed genes (fold change > |4| and p-value < 0.01), resulting in 746 contigs for females and 256 for males (Fig. 2A). Later, contigs selected through a volcano plot were clustered according to their normalized RPKM values and grouped into a heatmaps. For this purpose, the Manhattan distance was estimated and a complete

linkage was selected as a clustering strategy. Thus it was possible to determine transcriptional differences between each sex (Fig. 2B). Through this it was possible to identify up-regulated genes, such as the check details spermatogenic leucine zipper in males and vitellogenin in females ( Fig. 2C). Similar results were previously reported in sea lice (Caligus rogercresseyi) ( Farlora et al., 2014). The complete list of up-regulated genes in females and males are available in Table S2. Variant detection was conducted for the contigs exclusively found for each sex. The parameters were as follows: window length = 11, maximum gap and mismatch count = 2, minimum average quality of surrounding bases = 15, minimum quality of central base = 20, maximum coverage = 100, minimum coverage = 8, minimum variant frequency (%) = 35.0, and maximum expected variations (ploidy) = 2.

W przypadku obu szczepionek maksymalny efekt ochronny przed zaawansowanymi zmianami przedrakowymi można uzyskać, szczepiąc młode nastolatki przed kontaktem

z HPV (tab. 2). Badania immunogenności wykazały, że 8,4 roku po szczepieniu szczepionką Cervarix u 100% zaszczepionych nastolatek i młodych kobiet (w wieku 15–25 lat) w surowicy stwierdzono swoiste przeciwciała wobec HPV 16 i 18, a ich średnie stężenie utrzymywało się na stałym, wysokim poziomie (≥10 razy większym niż po naturalnym zakażeniu) przez cały okres obserwacji [37]. Na podstawie wyników analiz z zastosowaniem modelu matematycznego prognozowano, że takie stężenie przeciwciał przeciwko HPV 16 i 18 w surowicy po szczepieniu preparatem Cervarix utrzyma się nawet 50 lat [43]. Dasatinib mw Także w przypadku szczepionki Silgard stwierdzono, że średnio około 4 lata po rozpoczęciu szczepienia swoiste przeciwciała przeciwko HPV 16 były obecne u ponad 98% kobiet w UK-371804 wieku 16–26 lat, a średnie stężenie było około 3–4 razy większe niż po naturalnym zakażeniu [40]. Natomiast tylko 60% kobiet po tym okresie było seropozytywnych wobec HPV 18, a średnie stężenie swoistych przeciwciał w całej grupie

szybko po szczepieniu zmniejszyło się do wartości, takiej jak po naturalnym zakażeniu [20]. Jednak skuteczność kliniczna w zapobieganiu CIN związanej z HPV 18 utrzymywała się w przez cały ten okres na wysokim poziomie [40]. Immunogenność obu szczepionek jest znamiennie większa u młodych nastolatek w porównaniu ze starszymi grupami wiekowymi, zwłaszcza po 35. roku życia [36, 38, 41, 46]. W wieloośrodkowym badaniu klinicznym, w którym stosowano szczepionkę PtdIns(3,4)P2 Cervarix, stwierdzono ponad dwukrotnie wyższy poziom swoistych przeciwciał przeciwko HPV 16 i 18 u dziewcząt w wieku 10–14 lat w porównaniu z grupą 15–25

lat, zarówno po 7, jak i 48 miesiącach badania [39, 47]. Podobne zjawisko zaobserwowano również w badaniach szczepionki Silgard [46]. Indukcja dużego stężenia swoistych przeciwciał po szczepieniu w młodszej grupie wiekowej, które utrzymuje się na podobnym poziomie przez kilka lat (plateau), może być korzystnym prognostykiem trwałości utrzymywania się ochrony klinicznej przed zakażeniem HPV i rozwojem raka szyjki macicy [39]. Jednak to dalsze obserwacje i kolejne badania kliniczne każdej ze szczepionek osobno wykażą, czy i kiedy konieczne będzie podanie dawki przypominającej, aby utrzymać zadowalający poziom ochrony poszczepiennej przez długi czas.

,

2000), PD98059 purchase PLA2 clone A85/9-4 (Kanashiro et al., 2002), and hemorrhagin (Zn-metalloproteinase) clone 59/2-E4 (Barros et al., 1998) of B. atrox snake venom were cultured with DMEN-F12 medium, supplemented with 10% FCS and 10 μg/mL gentamicin. Each culture was expanded and 1 × 106 cells were inoculated i.p. in adult BALB/c mice previously i.p. injected with 400 μl mineral oil. After ten days, mice were euthanized by CO2 inhalation, and the ascitic fluid was collected by abdominal puncture. Monoclonal antibodies were purified with caprylic acid followed by ammonium sulfate precipitation (Steinbuch et al., 1970). Briefly, ascitic fluid was diluted 1:3 in 60 mM sodium acetate buffer, pH 4.0, and 0.4 mL caprylic acid was added under agitation for 30 min at room temperature for each 10 mL of ascitic fluid. The mixture was centrifuged at buy OSI-744 5000× g for 1 h and the supernatant was collected. After centrifugation, the pH of supernatant was adjusted to 7.0 and ammonium sulfate was added under agitation to achieve a 45% concentration (w/v), and the mixture allowed to stand at 4 °C overnight. Precipitates were recovered by centrifugation at 5000× g for 30 min, redissolved

and dialyzed against saline 0.9%, and immunochemically analyzed by SDS-PAGE and Western blot. Samples of dialyzed mAbs were subjected to 12% polyacrylamide gel electrophoresis (SDS-PAGE), according to the method described by Laemmli (1970) with modifications. The samples were dissolved in sample buffer (0.5 M Tris–HCl buffer, pH 6.8 plus 10% SDS, 10% 2-β-mercaptoethanol, and 0.5% bromophenol blue dye), boiled at 100 °C, loaded on 12% polyacrylamide gel, and run at 150 v. Protein bands were stained with Coomassie brilliant blue and subjected to computerized densitometric analysis (Bozzo and Retamal, 1991). Western blot was performed, according to a previously described method (Towbin et al., 1979).

Binding ability of the purified mAbs to the respective antigen was evaluated by ELISA test, according to the methodology described by Almeida et al. (1998). Briefly, B. atrox venom (10 μg/mL) or enriched fraction Methane monooxygenase of thrombin-like toxin (10 μg/mL) was diluted in 0.1 M carbonate/bicarbonate buffer (pH 9.6) and adsorbed to the ELISA plate. After a blocking step with gelatin, mAbs were diluted and added to wells. ELISA plates were incubated at 37 °C for 45 min followed by the addition of secondary antibody. The reaction was developed with o-phenylenediamine plus hydrogen peroxide, and color development was stopped with 50 μL 3 N H2SO4. Plates were read spectrophotometrically at 490 nm. Forty micrograms of myotoxic PLA2 from B. atrox venom, purified according to the method described by Kanashiro et al. (2002), were preincubated with 140 μg A85/9-4 mAb, and then aliquots of the mixtures were injected into the gastrocnemius muscle of five Swiss mice.

For calibration, we first measured the angular distributions of fetches with a step of 20° from the nautical charts for our study locations of Kõiguste and Matsi. However, it was difficult to assess the exact influences of islands, shoals and the coastline on waves, and the comparisons of results between the measured and modelled hourly time BMS-354825 mw series were not good enough. New distributions of fetches were created by maximizing the correlation coefficient and minimizing the root square error (RMSE) in the procedure, where the fetches are adjusted separately in all 20° sectors. The procedure also appears to enhance the fetches from the directions where the measured

wind forcing is restricted or distorted compared to the undisturbed wind properties at the wave measuring and modelling site. The calibration results are discussed in section 3.2. As the measuring period at Kõiguste was longer (221.2 days) than at Matsi (80.8 days), it included weather conditions over a larger range of variability. Variability ranges in sea level fluctuations measured as ‘instrument depth’ (1.23 m vs. 0.78 m, Table 1), salinity (1.18 vs. 0.83) and maximum wave heights (2.93 vs. 2.46 m) were also larger. Average properties of waves and currents at Kõiguste

were somewhat influenced by sea ice (Figure 2), which covered the measuring site for the first time at the end of December Epacadostat 2010. For a short period in February, the whole Gulf of Riga was ice-bound and ice forms of some kind were present until the end of April. Because of the proximity

to the coast, the measured currents tended to be polarized and modified by the coastline. Especially at Matsi, most of the velocity readings lay within two narrow directional intervals of 210–350 and 140–170 degrees: the v (S-N component) described 80–90% of the total variability ( Figure 3b). At Kõiguste, longshore (SW-NE) currents dominated as well, but as a result of the microfjord-like bottom topography, the directional scatter was considerably larger. Both currents and waves largely depended on wind conditions; no remarkable storm events occurred. At Matsi, however, both vertical distributions of currents ( Figure Cetuximab supplier 3a) and variations in thermohaline properties ( Figure 2h) indicated upwellingrelated changes in water column properties and coastal jets. It was discovered that, like the conditions on the Letipea Peninsula (Suursaar and Aps, 2007 and Suursaar, 2010) and some other specific Baltic locations (e.g. Jankowski, 2002 and Leppäranta and Myrberg, 2009), the straight coastal section near Matsi-Sõmeri is upwelling-prone when persistent northerly winds are blowing. Salinity increased and temperature decreased in summer (Figure 2h), and surprisingly high velocities were found in the surface layer.

Descoeur

et al. (2011) demonstrated these finding using TREK1–TRAAK null mice and use of the specific HCN inhibitor, ivabradine, which abolished the oxaliplatin-induced cold hypersensibility. An activation of slow axonal potassium (Kv7) channels reduces hyperexcitablity of axons in an in vitro model of oxaliplatin-induced acute neuropathy ( Sittl et al., 2010). TRPV1 is a capsaicin receptor that is activated by painful chemical stimuli, by noxious heat (activated at 42 °C) and inflammation. Transient receptor potential ankyrin 1 (TRPA1) co-localizes with TRPV1 in subpopulations of DRG neurons and it has a functional role in pain and neurogenic inflammation resulting from variety of compounds including irritant

chemicals, reactive oxygen and nitrogen species. It has been demonstrated that treatment with cisplatin and oxaliplatin buy Ibrutinib results in up-regulation of mRNA of TRPV1, TRPA1 and transient receptor potential melastatin 8 (TRPM8) in the cultured DRG neurons. Furthermore, up-regulation of TRPV1 and TRPA1 following in vivo treatment with cisplatin along with up-regulation of TRPA1 with in vivo treatment with oxaliplatin has also been reported. An up-regulation of TRPV1 and TRPA1 mRNA reflects an increase in TRPV1 and TRPA1 responsiveness in the nociceptors that contribute to the molecular mechanisms of the thermal hyperalgesia and mechanical allodynia observed in cisplatin-treated mice. Furthermore,

compared to the cisplatin-treated Olaparib manufacturer wild-type mice, cisplatin-treated TRPV1-null mice were ID-8 shown to develop only mechanical allodynia, but not the heat-evoked pain responses. It suggests that TRPV1 and TRPA1 could contribute to the development of thermal hyperalgesia and mechanical allodynia following cisplatin-induced painful neuropathy, and TRPV1 has a crucial role in cisplatin-induced thermal hyperalgesia in vivo ( Ta et al., 2010). The transient receptor potential vanilloid 4 (TRPV4) also plays a significant role in inducing mechanical hyperalgesia in paclitaxel-induced painful peripheral neuropathy (Alessandri-Haber et al., 2008). In models of painful peripheral neuropathy associated with vincristine and paclitaxel, mechanical hyperalgesia was reduced in TRPV4 knock-out mice and by spinal intrathecal administration of antisense oligodeoxynucleotides to TRPV4 (Alessandri-Haber et al., 2008). Oxaliplatin-induced cold allodynia is ascribed to enhanced sensitivity and expression levels of TRPM8 and TRPA1 (Gauchan et al., 2009a, Gauchan et al., 2009b, Gauchan et al., 2009c and Anand et al., 2010). TRPM8 is only expressed in the DRG and responds to innocuous cool and noxious cold (<15 °C) temperatures. Anand et al.

10. Despite the amount of uncertainty placed on these volume–weight calculations it appears

Veliparib cell line that published C-factors nonetheless underestimate the effects of urban forest cover in the region ( Fig. 11); however, in order to elucidate an appropriate C-value range for the area, an assessment of the contributions to the pond’s sediment budget unaffiliated with sheet and rill erosion from the surrounding landscape is required. The following two sources offer explanation for the inclusion of materials not accounted for by the USLE, which contribute to the overestimation of pond sedimentation due to inter-rill and rill processes: (1) sediment transported into the pond by anthropogenic means, and (2) gully erosion from surrounding hillsides. The pond is the final resting place for Epacadostat mw all materials derived from surrounding hillslopes and the footpath. A small source of error that could explain some of the variance between field and numerical model results is presented by the unknown factors associated with the upkeep of the footpath around the pond. Sand and gravel are replaced here on a regular basis as hillslope runoff not only carries materials from the slopes,

but also from the footpath into the pond. Evidence for this process is found in cores, which contained scattered pebbles found concentrated on the footpath. Since no records exist that would allow for quantification of this sediment source, the extent to which many these materials

offset measurements cannot be pursued; however, based on an assessment of collected sediment cores and a comparison of pond-sediment volume against path dimensions, it is assumed this contribution is negligible. It is likely that gullies are a significant contributor to the USLE model deviation; however, they provide an unquantified volume of sediment to the pond’s budget and pursuing their contribution from a process-oriented perspective would be time-consuming. It is estimated based on field reconnaissance of gully dimensions (width and depth) and their extent (derived from the flow-accumulation model) that the volume represented by gullies along the steep slopes north of the pond corresponds to ∼100 m3. This Gully-volume estimate is an order of magnitude smaller than the volume of sediment emplaced into the pond (∼6228 m3) and therefore would do little to close the gap between USLE model estimates of inter-rill and rill erosion and quantified pond sedimentation (Fig. 11). Regardless of how much gullying and anthropogenic contributions may add to the pond’s sediment budget, evidence suggests that urban forest cover promotes high erosion rates, which translate to high sediment flux and deposition within the pond. This is a function of the absent sediment storage anywhere along route within the watershed (Fig. 3); the study area thus provides a suitable location for a qualitative assessment of the C-value for this land-cover type.