In contrast, philanthotoxins (from the digger wasp Philanthus tri

In contrast, philanthotoxins (from the digger wasp Philanthus triangulum) and orb web spider polyamines can cause neuromuscular blockade by blocking nicotinic channels ( Rozental et al., 1989). We have previously reported the general properties and composition of venom from the Brazilian theraphosid spider Vitalius dubius Mello-Leitão 1923 ( Rocha-e-Silva et al., 2009a, b). In this work, we describe the neuromuscular activity of this venom and of a low molecular mass component capable of blocking vertebrate motor endplate cholinergic

nicotinic receptors. Male Swiss GSK2126458 price white mice (25–30 g) were obtained from the Multidisciplinary Center for Biological Investigation (CEMIB) at UNICAMP and male HY-LINE W36 chicks (4–8 days old) were supplied by Granja Globo Aves Agrovícola Ltda (Mogi Mirim, SP, Brazil). The animals were housed at 23 ± 2 °C on a 12 h light/dark cycle with access to food and water ad libitum. The animal experiments described here were approved by an institutional Committee for Ethics in Animal

Use (CEUA/UNICAMP, protocol no. 1587-1) and were in agreement with the Ethical Principles for Animal Research established by the Brazilian Society of Laboratory Animal Science (SBCAL). Venom was collected from male and female V. dubius by electrical stimulation ( Rocha-e-Silva et al., 2009b) into Eppendorf tubes covered with Parafilm® to avoid contamination with saliva. All reagents and salts for organ bath solutions were purchased from JT Baker (Center Valley, PA, USA) and drugs Enzalutamide price were from Sigma Chemical Co. (St. Louis, MO, USA) or JT Baker. Male chicks were killed with isoflurane

inhalation and the biventer cervicis muscles were removed (Ginsborg and Warriner, 1960) and mounted under a tension of 1 g/cm in a 5 mL organ bath containing warmed (37 °C), aerated (95% O2 + 5% CO2) Krebs solution of the following composition (in mM): NaCl 118.7, KCl 4.7, CaCl2 1.8, NaHCO3 25, MgSO4 Obatoclax Mesylate (GX15-070) 1.17, KH2PO4 1.17 and glucose 11.65, pH 7.5. A bipolar platinum ring electrode was placed around the tendon within which runs the nerve trunk supplying the muscle. Field stimulation was done with a Grass S48 stimulator (0.1 Hz, 0.2 ms, 4–6 V). Muscle contractions and contractures were recorded isometrically via a force-displacement transducer coupled to a PowerLab ML866/P digital myographic system (ADInstruments Pty. Ltd., Sydney, Australia). Contractures to exogenously applied acetylcholine (ACh, 110 μM) and KCl (20 mM) were obtained in the absence of field stimulation prior to treatments and at the end of the experiment, as a test for the presence of myotoxic and neurotoxic activities (Harvey et al., 1994). The preparations were allowed to stabilize for at least 20 min before the addition of ACh or KCl. Venom was added to the bath and changes in the contractile responses were monitored for 60 min for crude venom and up to 120 min for fractions.

Oceanographic modelling indicates a large proportion of floating

Oceanographic modelling indicates a large proportion of floating debris reaching the ocean will accumulate in gyres – the centre of vast anti-cyclonic, sub-tropical ocean currents. Using satellite-tracked “drifters” placed throughout the South Pacific ocean, Martinez et al. (2009) mapped the average trajectories of ocean currents, drift and eddies over time, the team found that, whilst some trackers were caught in near-shore currents, the majority fed into the south Pacific gyre from where they could not easily escape (Law et al.,

2010 and Martinez et al., 2009). Lagrangian drifters have also been used in a more recent study, indicating a high proportion of floating marine debris will end up in ocean gyres (Maximenko et al., in press). Data accumulated from over 6,000 plankton selleck kinase inhibitor tows conducted between 1986 and 2008 in the North Atlantic Ocean and Caribbean Sea, found plastic in 60% of the samples (Law et al., 2010). Mapping the plastic concentrations of each SB203580 transect, Law et al. (2010) revealed distinct spatial patterns of plastic in these areas, with highest concentrations

(83% of total plastic sampled) found in sub-tropical latitudes. The highest concentration was mapped to the North Atlantic gyre, with 20, 328 (±2, 324) pieces/km2. Due to the concentrations of plastic found it was impossible to determine the sources of such debris, but use of trackers suggested much of the eastern seaboard of the US fed into the gyre, taking debris 60 days on average to reach the gyre sited over 1,000 km away. Even higher plastic concentrations have been recorded in the North Pacific gyre: conducting 11 isothipendyl transects using a 333 μm manta-trawl, Moore et al. (2001) identified plastics in the majority of their

tows, with an average density of 334,271 plastic fragments/km2. Such work has led to significant media attention, with the North Pacific gyre being described “plastic soup” and coined as the “great Pacific garbage patch” (Kaiser, 2010). Plastics consist of many different polymers and, depending on their composition, density and shape, can be buoyant, neutrally-buoyant or sink. As such, microplastics may be found throughout the water column. Low-density microplastics are predominantly found in the sea-surface microlayer, as documented by numerous studies presenting data from surface trawls (Derraik, 2002 and Gregory, 1996). However, there is evidence that their position in the water column can vary: in estuarine habitats, low-density plastics, such as polypropylene and polyethylene, will be submerged if they meet water fronts. Furthermore, there is growing evidence that the attachment of fouling organisms can cause buoyant microplastics to sink (Barnes et al., 2009, Browne et al., 2010, Derraik, 2002 and Thompson et al., 2004). Plastic debris in the marine environment can rapidly accumulate microbial biofilms, which further permit the colonisation of algae and invertebrates on the plastics’ surface, thus increasing the density of the particle (Andrady, 2011).

A single influenza B virus isolated from a participant during 200

A single influenza B virus isolated from a participant during 2008, and propagated in MDCK cells was used to assess serum for both the first and second seasons. The virus had GDC-0068 ic50 a titer of 320 with B/Wisconsin/1/2010 (Yamagata) reference antisera and of <10 with B/Brisbane/60/2008

(Victoria) antisera. A reference antigen supplied by WHO (A/California/7/2009(H1N1)-like) was used to assess season 3/pandemic plasma. The HI titer was read as the reciprocal of the highest serum dilution causing complete inhibition of agglutination, partial agglutination was not scored as inhibition of agglutination. If there was no inhibition of HI at the highest serum concentration (1:10 dilution) the titer was designated as 5. Only one sample had a titer >1280 and this was not

adjusted. Influenza infection’ was defined as either the detection of influenza RNA in a swab sample by RT-PCR or a four fold or greater rise in HI titer, with a second titer of at least 40. Participants were excluded from analysis of each season if they were not present for ILI surveillance during the periods of Selleckchem Gefitinib confirmed influenza transmission or if paired-plasma were not collected. Additionally, participants were excluded from the analysis of effect of infection in one season on infection in subsequent season if they had not been available or fully assessed for infection in both seasons. The risk of an infection was modeled as depending on

the (log2-transformed) pre-season titer using a marginal logistic regression model, which takes into account potential household clustering. Unadjusted titer effects and titer effects adjusted for age (modeled as a natural cubic spline with 3 degrees of freedom and knots at 10 and 20 years) were calculated. We also tested for potential non-linear effects of the log2-titer on outcome by additionally including a quadratic term into the model and for titer–age interactions. The risk of infection was also modeled as depending on infection in the preceding season with each strain that did not induce HI antibodies (i.e. prior heterologous infections). As above, marginal logistic regression was used to account for potential household clustering and results adjusted for effects of age and pre-season HI titer. Statistical analyses Phosphoprotein phosphatase were performed with the statistical software R version 2.15.0 (R foundation for Statistical Computing, Vienna, Austria) and the companion R package geepack version 1.1-6. A detailed description of the cohort and of the infections and illnesses detected has been presented previously.21 In brief, 940 individuals were studied for three consecutive influenza seasons, from December 2007 through April 2010, resulting in 1793 person-seasons of influenza surveillance. The age of participants ranged from <1 to 90 years and none had ever received influenza vaccination.

To examine further the mode of action of this toxin and according

To examine further the mode of action of this toxin and according to the results presented just above, whole-cell voltage-clamp studies were carried out on the voltage-dependent inward sodium current (Lapied et al., 1990). Fig. 6 shows

representative inward sodium current traces elicited by a 30 ms depolarizing pulse applied to −10 mV from a holding potential of −90 mV, in control and after 24 min toxin application. Ipilimumab ic50 Application of μ-TRTX-An1a (100 nM) reduced the maximum amplitude of the sodium current by about 40% without affecting either time-to-peak or inactivation of the sodium current. The peak current–voltage relationship was illustrated in Fig. 6. This shows that the current started to activate at about −40 mV, reached a maximum

amplitude at about −15 mV and decreased to an extrapolated reversal potential of about +45 mV, a value which was very close to the Nernstian equilibrium potential for sodium ions. As shown in Fig. 6, μ-TRTX-An1a (100 nM) reduced the maximum current amplitude at all potentials tested. The potential at which the current was at its maximum and the reversal potential were all unaffected. Altogether, these electrophysiological effects clearly indicated that μ-TRTX-An1a was active upon more than one molecular target, being therefore a promiscuous toxin. Based on these results, it was tempting to suggest that the toxin could affect both voltage-dependent Alectinib concentration sodium currents and background sodium currents known to be 1) involved in the maintenance of the DUM neuron resting membrane potential at a relatively positive value (i.e., −50 mV) and 2) affected by scorpion toxins ( Lapied et al., 1999;

Grolleau et al., 2006). Furthermore, the toxin-induced increase the of the spontaneous firing frequency could result from an additional effect of μ-TRTX-An1a, particularly on voltage-dependent channels involved in the slow depolarizing phase during which the threshold of action potential is reached ( Grolleau and Lapied, 2000). Among voltage-dependent currents underlying this pacemaker potential, the low-voltage-activated transient and maintained calcium currents together with the maintained low-voltage-activated current permeable to both sodium and calcium ( Grolleau and Lapied, 1996; Defaix and Lapied, 2005) could be also targeted directly and/or indirectly (via changes in intracellular calcium concentration, for instance) by this toxin. In the present work, however, we were not able to investigate deeply the activity of μ-TRTX-An1a on other ion currents, nor to investigate the dose–response effect for its activity on DUM neuron electrical activity due to the limited amount of toxin available. Therefore, these aspects can be seen as perspectives for the continuation of this research.

Previous cross-sectional comparisons and short-term training stud

Previous cross-sectional comparisons and short-term training studies of the elderly generally support our viewpoint. Shinkai et al. (1998) saw little difference in CD3+, CD4+, CD8+, CD16+ or CD19+

counts between aerobically active and inactive elderly non-smokers; very fit individuals showed a superior T cell proliferative response to both PHA, and pokeweed mitogen, but the mixed lymphocyte reaction was not enhanced, making it unlikely that their T cell effector function was enhanced. Likewise, Arai et al. (2006) found that in elderly men the proliferative response to PHA was enhanced by aerobic training. Nieman et al. (1993) also made a cross-sectional BYL719 in vitro comparison between fit and unfit women aged 67–85 years; the highly trained individuals had a 54% advantage of lytic activity and a 56% greater T cell proliferative response to PHA, but there were no inter-group differences in lymphocyte subset counts, and a 12-week training programme did not enhance either T cell function or resting NK cell activity in the sedentary group. Woods et al. (1999) also found no significant increase of NK activity with six months of aerobic training in elderly men. A dissident report from Crist et al. (1989) noted a 33% increase

in resting NK cell activity in seven elderly subjects following 16 weeks of aerobic training. There is even less evidence of a positive response of immune parameters to resistance training (Raso et al., 2007, Flynn et al., 1999 and Kapasi et al., 2003), although McFarlin et al. (2004) did observe some increase of NK cell activity. In reviewing available reports, Vemurafenib Bruunsgaard and Pedersen (2000) concluded that physical training programmes acceptable to an elderly population are unable to bring about any major restoration of the senescent immune system. Our observations have been based on fitness scores, rather than questionnaire estimates of habitual physical activity. Although fitness scores reflect both the genetic characteristics of an individual and his Chlormezanone or her habitual physical activity, this approach to classifying the activity habits

of the elderly avoids the problems of recent memory often encountered when using questionnaires in such populations. Our subjects were typical of most elderly people, not athletic and relatively unfit compared with some previously reported groups; as might be predicted from previous reports on the general elderly population, there was little evidence that lymphocyte counts and sub-sets, the proportion of naive and memory cells, NK cell sub-sets, co-stimulatory molecules, apoptotic markers and activation markers differed between the upper and lower halves of the fitness spectrum, whether this was assessed in terms of aerobic power or muscle strength. Furthermore, three subjects who were immunologically “at risk” had similar levels of fitness to the remainder of our subjects.

, 1994) After its transportation, LPC is rapidly converted into

, 1994). After its transportation, LPC is rapidly converted into different products by specific routes which are important to regulate LPC levels on such tissues (Illingworth and Portman, 1972). However, the real content and the presence of LPC in cells or tissues are difficult to accurately determine (Schilling et al., 2004).

But, in vitro experiments showed that values above 50 μM, LPC is considered toxic, since plasma membrane integrity is disturbed due to its detergent-like feature ( Masamune et al., 2001). In fact, LPC is an intriguing molecule and should be more investigated. Data from literature point out the participation of PKC pathway in the retina ganglion cells leading them to survive (Santos and Araujo, 2000 and de Rezende Corrêa et al., 2005). PKC enzymes have been the primary mechanisms implicated in several biological effects, but the molecular basis for such activation Pirfenidone order is poorly understood. So, the increased survival BIBF 1120 molecular weight of retinal ganglion cells induced by LM-PLA2-I as well as LPC showed to be dependent of PKC pathway since this effect was abolished in the presence of a PKC inhibitor (chelerythrine chloride). PKC comprises a family of serine/threonine kinases involved in different events of neuronal development, as proliferation, survival and apoptosis (Wooten, 1999). This family is divided into three groups; the conventional one (α, β,

γ) depends on calcium ion and is activated by diacylglycerol and phorbol ester; the atypical (ζ, λ) is calcium independent and is not activated by diacylglycerol or phorbol ester

while the novel class (δ, ɛ, η, θ) is also calcium independent, but it is activated by diacylglycerol or phorbol ester (Michie and Nakagawa, 2005 and Reyland, 2009). To evaluate the participation of classical PKC isoforms and calcium, BAPTA-AM was employed. As seen, the survival effect induced by LM-PLA2-I was not affected in the presence of BAPTA-AM. Thus, discarding the involvement of such group and the need of increase intracellular calcium levels on this trophic effect. These results, in part, are in contrast to Rigoni et al. (2007) and Montecucco and Rosseto (2008). They observed that the neurotoxic effect exhibited by taipoxin (a potent snake PLA2 neurotoxin isolated from Oxyuranus scutellatus) was dependent on the increase on intracellular calcium levels. However, we would like to emphasize Quisqualic acid that our data are quite different from these authors’ results; because taipoxin displayed toxic effects on neurons and LM-PLA2-I had trophic or protective effects on neuronal ganglion cells. Rottlerin (a PKCδ isoform inhibitor) abolished the LM-PLA2-I-induced survival. However, rottlerin is not enough to state that the LPC-induced survival upon ganglion cells occurs through PKCδ pathway, but we might infer or postulate that this finding indicates a possible involvement of such enzyme to enhance on LPC-induced retinal ganglion survival effect.


A Selleck Quizartinib similar relationship between the intensification of the AABW formation and anomalous warming in the Southern Ocean has been discussed by Swingedouw et al.

(2008). The Southern Ocean warming is considerably strenghtened when implementing the new TKE scheme (F4 simulation), bringing the simulation much closer to the observed climatology in this region (Fig. 2). However, accounting for this new parameterisation also unrealistically damps the amplitude of the SST seasonal cycle in particular in the Northern Hemisphere (Fig. 3). This major drawback largely arises from the strong summer surface cooling driven by a deeper mixing penetration. It led to the decision not to include this parameterisation in the final CMIP5 version of the coupled IPSL model. Note that this simulation also includes the new RGB light penetration scheme that presumably drives the anomalous subsurface cooling in the tropics, while warm anomalies at mid-latitudes are most likely driven by changes in the mixed layer physics, as shown below. In the F5_CMIP5 configuration, the new TKE scheme was removed and a modelled 3-dimensional distribution of

chlorophyll was used. It remains difficult to decipher the specific effect of each of these modifications. A mid-latitude subsurface cooling largely compensates the warm bias that was detected in F4, highlighting the cancellation of the effect of the new TKE scheme. The upper right panel displays the temperature differences between F5_CMIP5 and F3, which sheds a light on a dominant cooling in the upper 200 m in the tropics and in the upper 400 m Casein kinase 1 in the subtropics. Fulvestrant clinical trial This cooling is attributable to several modifications in light penetration scheme, precisely the RGB scheme and the 1-dimensional response to biophysical feedback to the light penetration set by a present-day chlorophyll climatology. The impact of interactive biology is further investigated in the following sections. Fig. 4 shows the climatological SST differences between CM5_piCtrl and CM5_piCtrl_noBio. The annual mean SST is colder

over most of the globe when using the interactive biogeochemistry module. The effect is weaker and even opposite in eastern equatorial areas and coastal upwelling regions, as well as along western boundary currents at mid-latitudes and Southern and Arctic Oceans while it is strongest in the centre of subtropical gyres. The root mean square averaged SST difference among the two runs amounts 0.14 K. The one-dimensional thermal adjustment of the ocean to the inclusion of biogeochemistry is expected to induce an anomalous surface warming in CM5_piCtrl as compared to CM5_piCtrl_noBio in eutrophic regions (i.e. in regions where the modelled chlorophyll concentration is higher than 0.05 mg/m3). Indeed, high chlorophyll concentrations amplify the thermal disequilibrium in the water column by trapping more heat at the surface of the ocean and cooling subsurface waters.

After weight loss intervention, we observed significant improveme

After weight loss intervention, we observed significant improvements in BM, BMI, body fat selleckchem mass (% and kg), visceral and subcutaneous fat, insulin concentration, HOMA-IR, QUICKI, total cholesterol and triglycerides. Indeed, short- and long-term interventions increased the free fat mass (%) (Table 1Table 1). Based on the adipokine and neuropeptide data, we verified increases in adiponectin concentration and adiponectin/leptin

ratio with a concomitant reduction in alpha-MSH concentration. The leptin concentration decreased, while the orexigenic factors (ghrelin and AgRP) increased after 1 year. When we analyzed the AgRP from 6 months to 1 year of intervention, a significant increase was observed (Table 2Table 2). After weight loss intervention, we observed significant improvements in BM, BMI, body fat mass (% and kg), visceral and subcutaneous fat, insulin concentration, HOMA-IR, QUICKI, total cholesterol and triglycerides, similar to the trend observed in the normoleptinemic patients. Only long-term (1 year) treatment

was able to promote a significant increase in free fat mass (%) (Table 1). The group with hyperleptinemia exhibited a significant increase in adiponectin, NPY concentration and adiponectin/leptin ratio as well as a reduction in leptin and NPY/AgRP ratio with short-term intervention. After one year, this group presented a significant increase in adiponectin/leptin ratio and in AgRP concentration, Ribociclib cost with a reduction in the NPY/AgRP ratio (Table 2). Rutecarpine It is important to note that hyperleptinemic patients presented lower adiponectin/leptin ratio, alpha-MSH and ghrelin concentration at baseline. Indeed, the NPY/AgRP ratio was higher compared with that observed in the non-hyperleptinemic group. We found positive correlations between leptin concentrations and BMI and body fat mass (%) at baseline,

in the entire group (Fig. 1a and b). On the other hand, the leptin concentrations were negatively correlated with free fat mass (%) and alpha-MSH (Fig. 2a and b). Negative correlations between adiponectin/leptin ratio and total cholesterol and LDL-c were confirmed at baseline only in hyperleptinemic patients (Fig. 3a and b). As shown in Table 3Table 3, stepwise multiple linear regression analysis was performed with leptin concentration as the dependent variable. α-MSH and body fat mass (%) were the independent predictors to explain leptin concentration in the present study. Obesity has been shown to cause resistance or reduced sensitivity to several hormones, including leptin and adiponectin. Obese individuals appear to have higher sympathetic nervous system (SNS) activity; however, the metabolic response to SNS stimulation appears reduced in this population. This finding suggests that, in obesity, any compensatory effect of the SNS on metabolism to increase energy expenditure may not occur, rendering weight loss more difficult [8] and [33].

However, as water is lost to the ice outside the cell, intracellu

However, as water is lost to the ice outside the cell, intracellular processes including those involved in RCH may become inactive ( Danks, 2000). In the aforementioned study, B. antarctica was frozen inoculatively at -5 °C over 1 h, but there was no indication of when the organism actually froze, and so it is possible that the RCH observed was accrued prior to the freezing event in this organism. In general, the capacity for RCH is a valuable ecophysiological response for invertebrates, by allowing them to adjust rapidly to sudden changes in temperature on a temporal and spatial scale (Powell and Bale, 2005 and Sinclair and Chown, 2006). However,

the temperatures which RCH protects against in summer acclimated AZD8055 ic50 E. murphyi are rarely, if ever, seen on Signy Island during the active season ( Davey selleck kinase inhibitor et al., 1992). In addition, Worland (2010) has shown that, following long-term

acclimation (4 d at −4 °C), larvae can survive to −20 °C, a temperature never experienced in their soil habitat on Signy Island. Thus, RCH may prove to be unnecessary even in winter. Accordingly, RCH may serve a greater purpose at sub-lethal temperatures, with the enhancement of survival under limiting conditions in this study simply denoting a by-product of the RCH response acting on sub-lethal characteristics (e.g. reproduction) at temperatures more frequently seen in nature. Sub-lethal effects have been recorded in a number of

studies. For example, in D. melanogaster, Shreve et al. (2004) demonstrated an improvement in courting and reproduction at 16 °C after RCH, while Kelty and Lee (1999) identified a lower critical thermal minimum (CTmin, temperature below which activity does not occur). A reduction in the CTmin was also noted in S. avenae after RCH ( Powell and Bale, 2006). An analogous response in E. murphyi would clearly be ecologically beneficial. For instance, by being able to feed and, subsequently, develop at lower temperatures, E. murphyi might be in a better position at the end of the short growing season (cf. Hawes et al., 2007). For the majority of animals, RCH is thought to ameliorate chilling 4��8C injury, via the maintenance of membrane fluidity (Lee et al., 2006a, Lee et al., 2006b, Teets et al., 2008 and Overgaard et al., 2005) and the inhibition of apoptosis (Yi et al., 2007 and Yi and Lee, 2011). This interpretation is supported, in part, by the current study. As there was no significant difference between the SCPs of rapidly cold hardened and non-rapidly cold hardened larvae, the mechanisms involved in the RCH response are unlikely to have been associated with freezing injury prevention processes that alter the SCP, such as the accumulation of antifreeze proteins (AFPs) and the augmentation of ice nucleating agents (INAs) (Bale, 2002).

Settlement plates can be deployed to assess whether the colonisin

Settlement plates can be deployed to assess whether the colonising community has the same species composition as the previous community and/or

set aside area. Genetic analysis comparing the fauna colonising artificial or newly-generated natural substrate to the original populations could enable the source of colonisers to be identified Selleckchem Ceritinib and the suitability of set aside areas to be assessed. The monitoring program needs to be implemented at suitable spatial and temporal scales (IMMS, 2011), although the appropriate length of long-term study required is at present unclear. Levels of natural variation need to be evaluated before any appreciable operations begin, in order to establish fluctuations that could, for example, be seasonal or related to changing chemical conditions. Also, following disturbance, succession of species composition and abundance is to be expected, and so any monitoring must span sufficient time. Recovery from natural disturbance at sites along the EPR (Lutz et al.,

1994 and Mullineaux et al., 2010) and Juan de Fuca Ridge (Tunnicliffe et al., 1997) and the rapid re-growth of deposits at Solwara 1 (Gwyther, 2008a) indicate that monitoring for a few years following the cessation of mining activities may be sufficient. However, experimental polymetallic nodule mining resulted in PI3K Inhibitor Library purchase disturbance to the benthic community assemblage for at least 26 years following mining activity (Miljutin et al., 2011), suggesting that in keeping with the precautionary principle, suitable long-term monitoring could be on the scale of decades rather than years. Monitoring programmes by themselves are all very well, but they need to be evaluated against pre-determined decision rules. The latter will be derived from management objectives, and involve a management response when a monitored parameter value exceeds a certain level. For example, mining may have to stop in an area if sediment plume deposition thicknesses exceed a certain Flucloronide depth. The design of baseline, impact and long-term monitoring studies also needs to consider the importance of replication to address the natural

environmental variability at SMS sites at both temporal and spatial scales. Ideally, this should utilise a design similar to BACI (before-after-control-impact, Green (1979)) or Beyond BACI (Underwood, 1991 and Underwood, 1992), with multiple unimpacted (control or set aside) and impacted (mined) sites (Collins et al., 2013a). However, BACI design at SMS sites will probably be asymmetrical with the potential for multiple unimpacted sites but only one impacted site (Underwood, 1991 and Underwood, 1992), as mining is likely to be concentrated at one site. There is also the question of cost. Coastal or shallow water impact studies may be able to investigate multiple sites but the logistics (time and cost) of investigating multiple sites in deep-sea SMS mining impact studies may be prohibitive.