g. causing conflicts between data in text and tables, usage of standard formats and names, and defined usage of referenced values and experimental methods. None of the authors

have any conflict of interest. The SABIO-RK project is financed by the Klaus Tschira Foundation (http://www.klaus-tschira-stiftung.de/), the German Federal Ministry of Education and Research (http://www.bmbf.de/) through Virtual Liver and SysMO-LAB (Systems Biology of Microorganisms), and the DFG LIS (http://www.dfg.de/) as part of the project Integrierte Immunoblot Umgebung. “
“Enzyme assays are performed to serve two different purposes: (i) to identify a special enzyme, to prove its presence or absence in a distinct specimen, like an organism or a tissue and (ii) to determine the amount of the enzyme in the sample. While for the first, find more the qualitative approach, a clear positive or negative result is sufficient, the second, the quantitative approach must deliver Alectinib solubility dmso data as exact as possible. A great advantage of enzymes is that they can

be identified by their catalysed reactions, in contrast to the other components of the cell, like functional proteins or nucleic acids, which must be determined by direct detection. During the enzyme reaction product accumulates in amounts exceeding by far the intrinsic enzyme concentration. However, the conclusion from the product formed back to the amount of enzyme in the sample comprises various difficulties and pitfalls. Procedures for enzyme assays are documented or cited in various standard books (Methods in Enzymology; Advances in Enzymology

and Related Areas of Molecular Biology; Methods of Enzymatic Analysis (Bergmeyer, 1983); Springer Handbook of Enzymes (Schomburg, 2009); Practical Enzymology (Bisswanger, 2011) and databases (ExPASy database, and Brenda database,), but even accurate observance DNA ligase gives no guarantee of an unequivocal outcome. The same assays performed independently under obviously identical conditions may yield quite different results. In fact, the enzyme activity depends on manifold factors and general understanding of the particular features of enzymes is required, which cannot be described in all details in protocols for special enzyme assays. The most important aspects to be considered for enzyme assays are the subject of this article. It was the merit of Leonor Michaelis and Maud Menten (Michaelis and Menten, 1913) to realize that the enzyme activity depends decisively on defined conditions with respect to temperature, pH, nature and strength of ions and enzyme assays can reliably only be compared, if such conditions are strictly regarded. Considering these conditions, it may appear a simple task to define general rules valid for all enzyme assays, but such an endeavour will fail because of the great diversity of enzymes and their features.

, 2006). The largest proportion of sequences fell into the E6 category (n = 49, mostly of the D49 type, but also including N, K, R and H49 proteins). Most of the E6 proteins are acidic (4 > pI > 5.5),

but a few are neutral or weakly basic (pI = 6.4–8.95), although all are within the range previously reported for E6 proteins. For additional variants at the 6th position (A, G, R, T, W), see Table Etoposide S1. Oxidation products (clearly distinguishable as double peaks differing by 16 Da) were frequently present. Among the 10 samples that had been fractionated, isolated isoforms were found to be up to 20% oxidised. These often formed minor peaks in the LC–ES–MS and were generally absent in the MALDI–TOF spectra. From the 132 venoms examined, at least 83 masses representing putative unique PLA2 isoforms were identified between 13,193 and 14,916 Da. Between two (Popeia sabahi, A202, Ovophis makazayazaya,

A87) and 10 (Viridovipera gumprechti, B475) isoforms were found in the 24 samples with both LC–ES and MALDI–TOF–MS data. Between 25 and 100% (mean 70.45%) of isoforms in individual venoms were detected using both methods. Most of the masses which did not occur in both types of spectra were present as minor peaks in LC–ES–MS. About 70% of isoforms detected were scored as a major or minor peak consistently in both analyses. There was no significant PLX-4720 mouse difference between repeat spectra of the same venom sample, or from venom samples taken at different times from the same individual, although the relative intensity

of different peaks and presence of absence of minor peaks were not consistent in some cases. Out of the 73 proteins inferred from the genomic sequences obtained in this study, 62 (c. 85%) had a putative match in the expressed venom ( Table S1). However, several isoforms with different amino-acid sequences have inferred masses that are within 2 Da of each other, which are difficult to discriminate using proteomic methods ( Table S1), even the more accurate LC–ES–MS. Only 23 (32%) inferred PLA2 proteins were matched to masses in the venom profile of the Cepharanthine same individual from which the genome sequence had been obtained, suggesting that selective expression may account for a large proportion of among-individual variation in venom profiles. However, it also indicates incomplete sampling of the PLA2 gene content of the genomes investigated. The application of saline-loaded discs of filter paper caused no haemorrhage and no obvious disturbance to the chick embryos. Discs loaded with B. jararaca venom exhibited concentration-dependant haemorrhage, with a threshold concentration of 1.0 μg in 2.0 μl. The area of haemorrhagic corona increased with venom concentration and was maximal at a concentration of 3 μg in 2.0 μl, while the time taken for the corona to form fell. From these data, a ranking of haemorrhagic potential was calculated ( Table 1).

A syngeneic methylcholanthrene-induced sarcoma (MCA) cell line was used as previously described [3]. It was cultivated at 37°C with 5% CO2 in 20 ml of Roswell Park Memorial Institute (RPMI) medium 1640 medium

containing glutaril, 10% FBS, and 1% penicillin/streptomycin (Invitrogen Corporation/Gibco/Life Technologies Ltd, Paisley, United Kingdom). This procedure was performed as described previously [3]. Briefly, animals were anesthetized Epacadostat in vitro by pentobarbital sodium (50 mg/kg), and oro-tracheal intubation was performed using a 16-gauge polyethylene Angiocath (Becton Dickinson, Sandy, UT). Animals were ventilated with a mixture of oxygen and isofluran (0.5%-2%, Forene; Abbott, Zug, Switzerland) using a tidal volume of 10 ml/kg and a respiratory rate of 75 to 90/min. A left-sided minithoracotomy was performed through the seventh intercostal space, and 0.1 ml of MCA cell solution containing

5 × 107 viable tumor cells was injected subpleurally into the left lower lobe using a 27-gauge needle [12]. The thoracotomy was closed layer by layer, and the endotracheal tube was removed. Treatment was initiated when the tumors had reached a size of approximately 4 to 6 mm in diameter (approximately 7 days) as previously described [13]. The animals were anesthetized, and a left-sided thoracotomy Fenbendazole was performed through the fourth intercostal space. The left lung was freed from its adhesions. A left

cervical incision was performed to cannulate Doramapimod the external jugular vein. Visudyne was dissolved in NaCl (0.9%) and glucose (5%) and injected at a dose of 0.0625 mg/kg. After 15 minutes, laser light was applied to the exposed lower lung at a wavelength of 689 nm by an optical fiber–based frontal light distributor (Medlight, Ecublens, Switzerland) coupled to a diode laser (4-W laser diode; Biolitec, Germany). Noncontact, nonthermal surface irradiation was performed to the tumor and the surrounding normal lung tissue with the incident laser beam directed perpendicular to the lung surface and centered on the tumor. The treatment spot had a diameter of 30 mm, and the treated area was exposed to an irradiance of 35 mW/cm2 and a light dose of 10 J/cm2 corresponding to a treatment time of approximately 5 minutes. The irradiances and the light doses were measured in real-time as previously described [7] and [12]. Immediately after laser light delivery, 400 μg of Liporubicin dissolved in 0.5 ml of 6% Hydroxyethyl Starch (HAES) was injected through the external jugular vein catheter. The time interval between Liporubicin administration and harvesting of the left lung (Liporubicin circulation time) was 60 minutes.

Dabei handelte es sich insbesondere um erhöhte Fe-Spiegel, Marker für oxidativen Stress und Lipidperoxidation in der Substantia nigra. Interessanterweise verhindert und verzögert die pharmakologische Chelation bei erhöhtem Eisenspiegel in einem Modell für MPTP-induzierte Neurotoxizität für PS die Degeneration dopaminerger Neuronen im Mittelhirn [93]. Diese Studien weisen auf einen möglichen neurotoxischen Beitrag von Fe und Mn zur Neuropathologie des PS hin. Andererseits wurde

vorgeschlagen, dass Zn, das als Kofaktor für eine Reihe von Enzymen dient, an normalen neurologischen Funktionen beteiligt ist, weil es in signifikanter Konzentration (10 μM) im Gehirn vorliegt [94]. Zwar ist ein Teil des Zn im Gehirn mit Proteinen click here assoziiert, der Neocortex und der Hippocampus enthalten jedoch eine beträchtliche Menge an chelierbarem Zn [94], [95], [96] and [97]. Die definierten physiologischen Funktionen von Zn sind derzeit noch unklar, es scheint

jedoch an der Stabilisierung glutamathaltiger Vesikel an der Synapse sekretorischer Zellen beteiligt zu sein [98] and [99]. Außerdem haben in-vitro-Exprimente ergeben, dass Zn die NMDA-induzierte Toxizität verringert [100]. Die intrazerebroventrikuläre Verabreichung von Zn bei Ratten verursacht jedoch eine durch epileptische Anfälle ausgelöste Neurodegeneration im Hippocampus [101]. Tatsächlich haben weitere Untersuchungen ebenfalls gezeigt, dass Mn und andere Metalle (z. B. Cu, Al, Zn) mit Daporinad Proteinen interagieren und die Bildung von Amyloidfibrillen und die Aggregation z. B. von Prionproteinen (PrP) und α-Synuclein fördern können [102]. Diese Proteine binden Metalle, was zur Änderung ihrer Konformation und Löslichkeit beiträgt und ihre Aggregation unterstützt [103], [104], [105], [106] and [107]. Die in-vitro-Analyse von PrP-Aggregaten

ergab jedoch, dass Mn die Aggregation unabhängig von der PrP-Metallbindungsstelle fördern kann [106]. Wie gezeigt wurde, bindet Cu bei AK mit hoher Affinität an Aβ und moduliert dessen Konformationszustand und Peptidlänge [108] and [109]. Durch weitere in-vitro-Untersuchungen wurde demonstriert, dass Aβ mit Fe und Zn interagiert, was die Amyloidbildung fördert. Nintedanib (BIBF 1120) Interessanterweise werden diese Resultate durch post mortem durchgeführte Untersuchungen an Gehirnen von AK-Patienten gestützt, bei denen im Neocortex ein signifikant erhöhter Fe- bzw. Zn-Spiegel sowie Ablagerung von Amyloid-Plaques festgestellt wurden [109]. Alle diese Untersuchungen weisen darauf hin, dass Interaktionen zwischen Metallen und PrP, α-Synuclein und Aβ-Protein zum Zelltod führen können, da durch diese Wechselwirkungen die Bildung fehlerhafter und toxischer Proteinaggregate gefördert wird. Darüber hinaus werden Redoxzyklen unter Beteiligung der Fenton- und der Haber-Weiss-Reaktion induziert, die zur Depletion zellulärer Antioxidantien, wie z. B.

Coker 9553, the most susceptible cultivar to common foliar diseases considered in the study, was generally statistically different from the other cultivars and it also provided www.selleckchem.com/ALK.html the highest average. However, it did not provide the highest average net return from treatment. In fact, overall, none of the cultivars produced

a net return from treatment that was statistically different from the other three cultivars considered. A probability analysis based on Bayesian inference was also conducted to further assess whether a preventive application of a relatively inexpensive foliar fungicide to winter wheat in Northeast Texas is likely to result in a yield gain necessary to cover or exceed fungicide application costs. A Bayesian probability analysis has the advantage of incorporating some of the uncertainty that is associated with the treatment means. However, it should be emphasized that the cultivar’s susceptibility, the timing of the fungicide applications, grain prices, and fungicide costs can influence the probability of a profitable fungicide application. When the plants were sprayed at approximately Feekes Growth stage 10, wheat price was $0.25/kg, and tebuconazole and its application cost were $17.29, our probability analysis indicated that positive overall net returns for the cultivars analyzed are likely, and that most of them have the potential

to produce a yield gain that would break even on the tebuconazole spraying decision. Based on these probability ABT-199 order results, it is recommended to apply a preventive application of tebuconazole. Foliar fungicides could be a particularly valuable tool managing winter wheat in regions of moderate to high disease pressure. Our study also made several contributions to the current literature review on the economics of fungicide applications in wheat production. First, the study contributes

with additional findings related to the economic effect of fungicide Protirelin applications to prevent fungal diseases on wheat production. Second, the study illustrates the applicability of a Bayesian inference approach in evaluating net returns from fungicide applications. Finally, our study assists wheat farmers in Northeast Texas with economic tools that can be used in formulating educated expectations about their spaying decision and future net returns. The study analyzed four red winter wheat cultivars (Magnolia, Terral LA841, Pioneer 25R47, Coker 9553), but due to data availability it was limited to two years (2011 and 2012). There were additional cultivars that were excluded from the analysis because they were not planted during 2011 and 2012. However, additional years can be analyzed when cultivars are grouped into categories. For instance, Thompson et al. (2014) were able to analyze data from 2005 to 2012 by grouping cultivars into three categories (resistant, intermediate, and susceptible cultivars) and by assuming that two different fungicides provide similar disease control.

, 2009 and Doyle et al., 2011). These plastic fragments constitute a frequently reported size inventory in many ingestion studies (Eriksson and Burton, 2003, Foekema et al., 2013 and Graham and Thompson, 2009). The size range of MP determines the potential impact of these contaminants on ecosystem biota (Mohamed Nor and Obbard, 2014). Dominance of smaller

particles increases the risks related to encounter frequency. S-MPPs were easily found in filter feeders in contrast to L-MPPs which were found frequently in carnivorous taxa (Foekema et al., 2013). The two research areas shared a similar composition of MP types. According to their shape, MP particles were categorized into four types: fibres, granules, plastic films and spherules. The fibres were the most common type, followed by granules and films. Spherules were the least common learn more type (Table 3). The click here similar share of MP types in the Yangtze Estuary and East China Sea indicated a possible MP flux from the river to the adjacent sea. Fibrous MPs seems to be most abundant in the marine environment (Wright et al., 2013). Being adjacent to the most highly populated region, the study areas are bound to accept large amounts of land-based debris. This is in accordance with Browne et al. (2011) who suggested that the majority of MP fibres found in the marine

environment may be derived from sewage as a consequence of laundering clothes. On the other hand, the fibres may derive from rope material. Heavy marine traffic and fishery activities in the study areas brought more discarded rope material (Andrady, Olopatadine 2011 and Thompson et al., 2004). Lacking identification of the polymer types, further speculation on the

origins of plastic particles cannot be made. The potential negative impacts of plastic particles ingested were proved to be associated with various particle shapes (Wright et al., 2013). If ingested, organisms inhabiting the study areas are vulnerable to the shape-related toxicity of fibrous MPs. Strikingly, spherules were rarely found in our study while commonly existing in water column samples (Moore et al., 2001 and Law et al., 2010). A decrease in spherules may suggest that industry initiatives have been useful in reducing the loss of pellets into the environment during transportation. Similar results have been reported in two other studies (Ivar do Sul et al., 2013b and Ryan, 2008). Transparent and coloured MPs were the majority of plastic items, with small fractions of white and black plastic items (Fig. 3). Prominence of transparent and coloured MP corresponds to the prevalence of clear plastics used in the plastic products, such as packaging, clothing and fishing line (Cole et al., 2014). The colours may potentially contribute to the likelihood of MP ingestion due to food resemblance, the prevalence of plastics with these colours in the environment and an actual colour preference by the biota (Costa et al., 2010, Shaw and Day, 1994, Verlis et al.

non-UK). No significant findings arose for these variables, thus they will not be discussed again. First, we looked at the two samples together, examining the perceived impacts of visits on the environment and on the visitor. We then explored any differences between coastal experts’ and coastal users’ ratings. To calculate the total perceived risk to the environment, perceived commonness of each activity was multiplied by perceived harmfulness (see

supplementary material for the individual means). As shown in Table 1, it was found that activities did significantly differ in terms of their perceived risk to the environment; with rock pooling, fishing 3-MA in vivo and crabbing seen to have the highest risk to the shore, and cycling, swimming and sunbathing/relaxing having the least. Qualitative data in response to if there was one visitor-related behaviour you would change in regard

to damage caused to rocky shore species or habitats, what would it be and why emphasised problematic activities and behaviours further. A total of 106 comments (25 from coastal experts, 81 from the non-expert sample) were collected. From their comments, three AC220 mouse prominent themes were found: Littering, lack of rock pooling ethics and general disturbance. Littering represented comments directly referring to the leaving of rubbish (e.g. generally, food-related, fishing, or dog fouling). For instance, “…The rubbish left behind is an eye sore and potentially dangerous to other visitors or the wildlife”. Lack of rock pooling ethics generally referred to acting in an inconsiderate manner in the rock pools (e.g. displaying general lack of knowledge, not turning boulders back, not returning organisms) that can lead to “…exposing animals and plants to the drying air is not good and will

change the ecology of a location in time”. The final theme, general disturbance, covers comments that addressed more generally Liothyronine Sodium the disturbance by visitors to the habitat and the wildlife such as from walking over the rocks or from rock pooling or crabbing, e.g. “…in terms of disturbing the habitat of shore creatures.” Littering behaviours were mentioned the most ( Table 2). All activities were perceived to have a positive impact on visitors’ mood, as all values were above the no change value of 3 for one-sample t-tests (all ps < 0.001; Table 3). Activities were found to differ from one another in terms of change in mood; as walking, wildlife watching and snorkelling were seen to have the most positive impact, whereas cycling, fossil hunting and jogging had the least positive impact ( Table 3). For the excitement scale, any values below 3 represent calming feelings, whilst values above 3 represent increased feelings of excitement. One-sample t-tests found that playing with the family, crabbing, snorkelling, rock pooling, fossil hunting and cycling were seen to make visitors feel more excited (all ps < 0.02).

The analysis of the texts selected for this review indicated that there are no studies directly associated to the factors which represent risk for pregnant women to search for late-term abortion after rape. However, seven studies have highlighted significant initiatives and procedures that can reduce risks and avoid late-term unsafe abortion (Table

1). The woman who seeks deliberate abortion may consider different reasons such as economic difficulties, health problems, neglect or lack of a partner, interference on the project life, conflict with society’s rules, or social vulnerability. In all cases, the common element is unwanted pregnancy, which makes the decision of abortion complex and multifactorial.5 Drezett et al. (1998)6 have assumed that the variability in gestational age of women seeking legal see more abortion could be related to difficulties in access to health services and barriers to the development of violence and pregnancy. However, other conditions may be associated, such as vulnerability

and limiting the autonomy of people with mental illness. It is also possible that crimes in which the click here perpetrator threatens the physical integrity of the victim or a family member, produce a similar effect. Mitchell et al. (2014)7 also showed that abortion knowledge and attitudes are not driven simply by age, religion or class, but rather a complex interplay that includes both social spaces and gender. Prevention of abortion morbidity and mortality among adolescents requires comprehensive sexuality and reproductive health education that includes factual distinctions between safe and unsafe abortion methods. The difference in the legalization of abortion across countries increases the complexity of the consequences of rape. The laws of each country determine the extent of the problem and dictate the rules and procedures viable, leaving health services act within the established limits. According to Kalonda (2012),8 from the politico-legal point of view, ending rape impunity

and decriminalizing abortion are recommended. Beta adrenergic receptor kinase Decriminalizing abortion give women choice and save victims and pregnant women from risks related to the pregnancy, a childbirth, or an eventual unsafe abortion. These risks increase the maternal mortality already high in Congo-Kinshasa (between 950 and 3,000 for 100,000 live births). After reviewing the laws of the 191 countries around the world for which information is available and categorizing them by legal indications, which include preservation of the woman’s life, health reasons, pregnancy due to sex offences, fetal impairment, socio-economic reasons, Boland (2010)9 concluded that while most countries may not decriminalise all abortions in the near future, especially second trimester abortions, less comprehensive legislative and regulatory reforms are possible.

Highly homologous to buy H 89 histones, they have potent, broad-spectrum activity against Gram-negative bacteria, water molds and parasites (Richards et al., 2001 and Fernandes et al., 2002). Another example is the antimicrobial peptide hipposin from the skin mucus of Atlantic halibut (Hippoglossus hippoglossus L.) derived from the histone H2A ( Birkemo et al., 2003). Other antimicrobial proteins isolated from fish and having other primary functions include apolipoproteins A-I and A-II, present in skin or serum of carp (Cyprinus carpio) and active against some fish bacterial pathogens ( Concha et al., 2004). These proteins with other well established

functions appear to be recruited to a second antimicrobial role in nature. In the present work we purified and identified the fraction of the P. cf henlei mucus responsible for antimicrobial activity against E. coli, M. luteus and C. tropicalis. The purified PcfHb exhibited a lower MIC against gram-negative bacteria and higher against gram-positive bacteria and fungi. The MIC values were in the same range as well-characterized peptide fragments from bovine hemoglobin ( Adje et al., 2011) and antimicrobial peptides including pardaxins and hipposins ( Oren and Shai, Alectinib cell line 1996 and Birkemo et al., 2003). Interestingly, the partial sequence alignment of PcfHb with several hemoglobin β-chain of different species,

demonstrated a high degree of conservation of certain amino acids ( Table 1). Some factors could explain the surprising antimicrobial activity of fragments of hemoglobin. One possibility is that the heme moiety

could act either as an iron chelator or as an oxidant, leading to damage of the bacterial and fungal cell walls. Parish et al. (2001) working with isolated chains of hemoglobin identified that the isolated DNA ligase β chain without heme exhibited activity on tested organisms, supporting the hypothesis that the heme plays no role in the antimicrobial activity and that subunit separation leads to enhanced activity. Thus, although the hemoglobin tetramer is only negligibly active against two gram-positive organism, the activity of the isolated β globin chain is greatly enhanced. In the case of β+heme, antimicrobial activity was observed against two of the bacterial targets but not on C. albicans. The results with isolated subunits indicate that tetramer dissociation exposes additional bioactive peptidic surfaces. Even though the tested microorganisms do not affect freshwater fish such as stingrays, proteins homologous to hemoglobin are also present in the microsomes of gill cells from a number of teleosts including Mozambique tilapia, Oreochromis mossambicus (Peters), rainbow trout, common carp, Cyprinus carpio L., European eel, Anguilla anguilla (L.), elephant fish, Gnathonemus petersii (Günther) ( Stekhoven et al., 2004) as well the presence of a family of AMPs derived from Hb-β present in the skin and gill epithelium of channel catfish ( Ullal and Noga, 2010).

planci; and (2) explore possible side-effects associated with the use of these chemicals, testing Birinapant for any evidence of disease or ill-health in other coral reef organisms (e.g., corals, fishes and other echinoderms) that feed on or are in close contact

with dying A. planci. A total of 397 adult A. planci specimens were collected at the Tandayag Marine Sanctuary in Amlan, Negros Oriental, central Philippines (9° 27′ 10.12″ N, 123° 14′ 14.81″ E) by local fishermen who were freediving up to 15 m depth and collected starfish using improvised bamboo tongs. Specimens were transported to the Institute of Environmental and Marine Sciences of Silliman University (SU-IEMS) in Dumaguete, Negros Oriental, Philippines and kept in 2 m3 concrete tanks with flow-through ambient seawater and left to acclimatize for 3 days. Weak and damaged individuals were discarded. Peptones, bile derivatives, TCBS, and yeast were tested to determine lethal doses (Table 1). Peptones used were bacteriological peptone, proteose peptone, special peptone, peptone EHCK, peptone 2400, and peptone 2382. Bacteriological peptone is mixed pancreatic and papaic digest Fluorouracil concentration of different animal proteins containing a wide molecular weight distribution of peptides. Proteose peptone is enzymatic digest of animal proteins with high content of low molecular weight proteoses used to create

an environment beneficial to the maintenance of virulence and the elaboration of bacterial by-products. Special peptone is prepared from meat, plant and yeast digest which contains the widest spectrum of peptide structures available in any peptone. Peptones EHCK, 2400 and 2382 are pancreatic digest of casein and whey (milk derivatives) with different molecular weights. Oxgall is dehydrated fresh bovine bile while bile salts N3 Phospholipase D1 may be effective at less than one-third of the normal concentration of bile salts and are usually added as selective inhibitory agents in culture media. Ten 95-l plastic bins were placed inside a large concrete tank, which served as a water bath. The depth

of seawater in the concrete tank was set to 20 cm, about half of the depth inside the plastic bins to maintain ambient temperature (28.5 °C) within each individual bin. Each plastic bin was supplied with constant flow of fresh seawater (40 l/min). Ten seemingly healthy sea stars (15–25 cm) were haphazardly selected from the stock and placed in individual bins. Ten ml of each chemical at different concentrations (Table 1) were injected to each sea star using a 21-gauge syringe. There were 10 replicates for each chemical tested except for bacteriological peptone (200 g l−1), peptone EHCK (100 g l−1), peptone 2400 (200 g l−1), and peptone 2382 (200 g l−1), where only 5 replicates were used because of the inefficacy and variability in results displayed by those types of peptones. The reaction of sea stars was evaluated at 1 h, 8 h, 24, and 48 h after injection.