Ecotoxicological studies of direct and indirect genotoxicity with Artemia: a integrative review.

Artemia is a brine shrimp genus adapted to extreme habitats like ranges salinity from 5-25 g/L and in temperatures from 9 to 35 °C. It is widely distributed and used as an environmental quality biomarker. Artemia franciscana and Artemia salina species are commonly used in ecotoxicological studies and genotoxicity assays due to their short life cycle, high fecundity rate, easy culture, and availability. Thus, considering the importance of these tests in ecotoxicological studies, the present study aimed to present Artemia genus as a biological model in genotoxicity research. To this end, we reviewed the literature, analyzing data published until July 2023 in the Web of Science, SCOPUS, Embase, and PubMed databases. After screening, we selected 34 studies in which the genotoxicity of Artemia for various substances. This review presents the variability of the experimental planning of assays and biomarkers in genotoxicity using Artemia genus as a biological model for ecotoxicological studies and show the possibility of monitoring biochemical alterations and genetic damage effects. Also highlight innovative technologies such as transcriptomic and metabolomic analysis, as well as studies over successive generations to identify changes in DNA and consequently in gene expression.

Parasitism in Rattus rattus and sympatric Achatina fulica by Angiostrongylus cantonensis in an urban park in southeast Brazil.

In this study, rodents (Rattus rattus) and mollusks (Achatina fulica) were captured in a small forest located in a large metropolitan city in Brazil, and they were examined to investigate possible parasitism by Angiostrongylus cantonensis. The parasites were recovered as helminths from the pulmonary arteries of the synanthropic rodents and as third-stage larvae (with Metastrongylidae family characteristics) from the mollusks. To confirm the species, these larvae were used to experimentally infect Rattus norvegicus for the posterior recovery of adult helminths. To identify the adult helminths, morphological, morphometric, molecular, and phylogenetic techniques were employed. Furthermore, we also characterized the histological lesions associated with parasitism in naturally infected definitive hosts. Our results demonstrated the occurrence of a natural life cycle of A. cantonensis (with the presence of adult helminths) in definitive hosts, Rattus rattus, and third-stage larvae in an intermediate host, A. fulica. In free-living rodents, lesions of granulomatous pneumonia in the lungs and meningitis in the brain were also found. These results warn of the risk of accidental transmission of A. cantonensis to human residents around the park because of the extensive interaction among the fauna of the park, domestic animals, and the surrounding human population.

Prognostic assessment for chronic myelomonocytic leukemia in the context of the World Health Organization 2016 proposal: a multicenter study of 280 patients.

Knowledge on chronic myelomonocytic leukemia (CMML) patients from Argentina and Brazil is limited. Our series of 280 patients depicted an older age at diagnosis (median 72 years old), 26% of aberrant karyotypes, and a prevalence of myelodysplastic (60%) and CMML-0 subtypes (56%). The median overall survival (OS) was 48.2 months for patients in CMML-0 (Ref.), 24.7 months for those in CMML-1 (HR = 2.0, p = 0.001), and 8.8 months for patients in CMML-2 (HR = 4.6, p < 0.001). In the CMML-0 category, median OS were different between myelodysplastic and myeloproliferative subtypes (63.7 vs 21.2 months, p < 0.001); however, no differences were observed within CMML-1 and CMML-2 subtypes (24.7 vs 23.7 months, p = 0.540, and 9.1 vs 8.2 months, p = 0.160). The prognostic impact of 24 variables and 7 prognostic systems was adjusted to the WHO 2016 after validating their usefulness. Multivariate analysis were performed, and the final model revealed Hb ≥ 8 -< 10g/dL (HR 1.7), Hb < 8g/dL (HR 2.8), poor karyotypes (HR 2.1), WHO 2016-CMML-1 (HR 2.1), and CMML-2 (HR 3.5) as independent adverse clinical parameters in our cohort with a borderline influence of platelets count < 50 × 109/L (HR 1.4). We could validate several scoring systems, the WHO 2016 proposal and its prognostic capability, along with accessible covariates, on predicting the outcome in our series of CMML patients from Latin America.

Plants of the Cerrado naturally selected by grazing sheep may have potential for inhibiting development of Haemonchus contortus larva.

Plant species naturally selected by sheep grazing in the Cerrado region of Brazil were assessed in vitro for activity against Haemonchus contortus. One year of observations showed the plant families in the region exhibiting greatest richness to be Fabaceae, Rubiaceae, Malpighiaceae, Bignoniaceae, Myrtaceae, and Annonaceae. Nine species commonly selected by grazing sheep showed variation in the selectivity index with respect to the dry and rainy seasons. Coproculture was conducted in five replicates of 11 treatments: ivermectin, distilled water, or dehydrated leaves of nine selected plant species administered at 333.3 mg g(-1) fecal culture. The dried powder of Piptadenia viridiflora and Ximenia americana leaves significantly reduced the number of infective larvae compared to the distilled water control. These species showed efficacy of over 85 % despite low concentrations of proanthocyanidin. High-performance liquid chromatography analyses of extracts of these plants showed major peaks of UV spectra characteristic of flavonoids. Those naturally selected plant species with high antihelminthic efficacy show promise for use in diet as an alternative control of H. contortus in sheep.

The effect of Momordica charantia extract on sheep infected with Haemonchus contortus.

Bitter melon (Momordica charantia) has several biological properties including anthelmintic activity. Presented in vivo study evaluated the anthelmintic activity of the hydroalcoholic extract of M. charantia Presented in vivo study (HEMC) and its effect on animal performance, carcass characteristics and lamb meat quality in animals infected by Haemonchus contortus. The HEMC demonstrated effectiveness against the parasite. Lambs treated with HEMC showed a significant reduction in egg count per gram of faeces and in the number of parasites in the abomasum. However, nutrient intake decreased with increasing doses of HEMC, leading to reductions in final live weight and carcass weight. No significant effects were observed on meat quality. Therefore, it is concluded that HEMC presents anthelmintic action, but it also impairs animal performance, highlighting the need to assess the benefits and impacts of HEMC use for H. contortus control.

Using freshwater snail Biomphalaria glabrata (Say, 1818) as a biological model for ecotoxicology studies: a systematic review.

Over time, a growing increase in human pollutants in the aquatic environment has been observed. The global presence of residues in water bodies reinforces the need to develop improved methods to detect them and evaluate their ecotoxicological effects in aquatic environments. Thus, this study aimed to present the main assays using Biomphalaria glabrata as a biological model for ecotoxicological studies. We performed a systematic literature review with data published up to June 2022 on the Web of Science, SCOPUS, Science Direct, PubMed, and SciELO databases. Thirty studies were selected for this review after screening. Biomphalaria glabrata has been studied as an ecotoxicological model for different substances through toxicity, embryotoxicity, cytotoxicity, genotoxicity, and bioaccumulation assays. Studies evaluating the impact of B. glabrata exposure to several substances have reported effects on their offspring, as well as toxicity and behavioral and reproductive effects. This review presents various assays using B. glabrata as a biological model for ecotoxicological studies. The use of a representative species of ecosystems from tropical regions is a necessary tool for tropical environmental monitoring. It was observed that the freshwater snail B. glabrata was effective for the evaluation of the ecotoxicity of several types of chemical substances, but further studies are needed to standardize the model.

Toxicological evaluation of antiretroviral Tenofovir Disoproxil Fumarate on the mollusk Biomphalaria glabrata and its hemocytes.

Tenofovir disoproxil fumarate (TDF) is a drug used in HIV treatment, and several studies have detected its presence in surface water. Furthermore, more information on its environmental impact is needed in the scientific literature. Thus, due to the lack of data on the impact of this drug, and its presence in different waters of the world, this work aimed to evaluate the potential toxicological effects of TDF on the mollusk Biomphalaria glabrata in vivo and in vitro. For in vitro analysis, hemocytes were exposed to different drug concentrations for 1 h and evaluated for feasibility, and phagocytic and metabolic activity. The in vivo analysis consisted of the exposure of groups of five mollusks, in triplicate, at the same drug concentrations for 72 h and 21 days, evaluating mortality, and mollusk and hemolymph behavior. Although the exposure of the mollusk to TDF did not reduce its survival, however it was toxic to its hemocytes. Even if toxicity was identified on the mollusk and its hemocytes initially, further studies should be conducted to understand the effects of this residue on the environment and different life stages of the mollusk because, per the Globally Harmonized System of Classification and Labeling of Chemicals, for aquatic ecosystems, the results obtained were classified as toxic (EC50% 2.65 [1.98; 5.29] mg/L) and could cause unfeasibility in hemocytes at concentrations below 10 mg/l.

Anthelmintic resistance in sheep in the semiarid region of Minas Gerais, Brazil.

Anthelmintic efficacy was evaluated among sheep that had become naturally infected with gastrointestinal nematodes in 17 flocks located in the semiarid region of Minas Gerais, Brazil. Feces were collected individually from 1021 hairy sheep to determine the number of eggs per gram of feces (EPG) and for coprocultures to identify nematode genera the nematodes. Only the animals that presented EPG counts greater than or equal to 200 were included in the study (totaling 381 sheep). The animals were divided into three treatment groups: albendazole, ivermectin and levamisole. Fourteen days after the administration of anthelmintics, fecal samples were taken from all animals. In each flock, the pre-treatment and post-treatment arithmetic mean EPG were used to calculate the efficacy (FECR) for each of the treatment groups and the lower 95% confidence limit. Data were analyzed with the "eggCounts 2.3" package in RStudio, using a Bayesian model for paired design. The anthelmintics were classified as being efficacious (when the FECR was both equal to or above 95% and the lower 95% confidence limit was equal to or above 90%) or as encountering anthelmintic resistance (when the FECR was below 95% and the lower 95% confidence limit was below 90%) or inconclusive (when none of the other criteria were fulfilled). Albendazole and ivermectin were not effective in any of the flocks. Levamisole was effective against gastrointestinal nematodes in 25% of the flocks studied. Haemonchus, Trichostrongylus and Oesophagostomum genera were identified in this study in a semiarid region of Minas Gerais, Brazil. The genus Haemonchus was the most prevalent, followed by Trichostrongylus and Oesophagostomum. After anthelmintic treatment, the most prevalent genus was Haemonchus, followed by Trichostrongylus; the genus Oesophagostomum was not detected. The highest percentage of Haemonchus larvae was observed after treatment with ivermectin, followed by albendazole and levamisole. This study revealed the existence of gastrointestinal nematodes in sheep that present multiple resistance to all three main classes of anthelmintic drugs.

Effect of temperature on activity of third-stage larvae of Angiostrongylus vasorum.

To evaluate the effect of temperature on the activity and mortality of the L3 of Angiostrongylus vasorum, 1,500 L3 were isolated from experimentally infected snails and distributed into five equal groups. Three groups were incubated at 37°C, 27°C, and 5°C. The remaining two groups were incubated at 27°C and 5°C for 10 days, at which time the temperature for the 27°C group was reduced to 5°C and the 5°C group increased to 27°C. Larva activity was observed daily and inactive larvae were removed. At 37°C, larvae survived up to 8 days. At 27°C, larvae were active until day 6. When subjected to a reduction in temperature from 27°C to 5°C beginning on day 10, the number of active larvae increased until day 13. Only on day 17 did the number of active larvae decline to zero. At 5°C, larvae remained active until day 15, surviving to 24 days. When temperature was increased from 5°C to 27°C beginning on day 10, larvae were found active until day 12 and maintained an intermediate level of activity to day 21. Survival of larvae was greater at lower temperatures, while high temperatures were associated with higher mortality.

In vitro activity of a serine protease from Monacrosporium thaumasium fungus against first-stage larvae of Angiostrongylus vasorum.

A serine protease from the nematophagous fungus Monacrosporium thaumasium (NF34a) was purified, partially characterized and tested in vitro in control of the first larval stage of Angiostrongylus vasorum. NF34a grew in liquid culture medium, producing its crude extract that was purified by ion exchange chromatography. The fractions with high protease activity were collected in a pool, and elution of proteases was monitored by enzymatic assay and protein content. Purification steps were monitored by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Protease activity was determined under different pH and temperature conditions, and the inhibitor effects of metal ions and phenylmethylsulfonyl fluoride (PMSF) were assessed. In an experimental test, the infection process of NF34a on first-stage larvae of A. vasorum was investigated. A purified serine protease (Mt1) was identified, with an approximate molecular mass of 40 kDa and apparent homogeneity in SDS-PAGE, having optimal activity at pH 7.0 to 8.0 and temperature of 60°C. Mg(2+) and Zn(2+) partially inhibited the activity of Mt1 while PMSF inhibited it completely. Mt1 production was observed when NF34a was grown using first-stage larvae of A. vasorum as the only source of carbon and nitrogen. These results show that the enzyme may have a possible role in the infection process of the larvae. In the in vitro test of applicability against A. vasorum L(1), we observed a reduction in the number of larvae of 23.9% (p < 0.05) in the group treated with Mt1 compared with the control group. However, even this low reduction demonstrates that the Mt1 is important in the infection process.

Anthelmintic efficacy in strongyles of horses in Northern Minas Gerais, Brazil.

The intensive use of anthelmintics has resulted in resistant parasite populations in horses. The objective of this trial was to evaluate the anthelmintic efficacies of the anthelmintics fenbendazole, ivermectin and abamectin in 24 horse farms in Northern Minas Gerais. Egg counts per gram of faeces (EPG) were performed individually in 619 animals. Animals presenting EPG counts greater than or equal to 150 were used in the tests on faecal egg count reduction (FECR), totalling 436 equines. These animals received the anthelmintics, fenbendazole, ivermectin, and abamectin. Faeces were collected 14 days after the administration of anthelmintics to perform the EPG. Pre- and post-treatment EPG counts were used to calculate the FECR for each anthelmintic group, and faecal culture was used to identificy of the strongyles. The resistance status was evaluated based on the FECR and LCL95%. Fenbendazole was effective in 11 (45.8%) of the horse farms. Ivermectin was effective in 17 (77.3%) and abamectin in 17 (74%) of the farms; side-resistance was detected in 3 (12.5%) of the farms. Intestinal strongyle resistance to anthelmintics was observed in 14 (58.3%) of the farms. Cyathostomin larvae were found in 100% of the farms, Strongylus vulgaris in 13 (54.2%), and S. equinus in 3 (12.5%). Only cyathostomins larvae were detected post-treatment with ivermectin and abamectin.

Dynamics of the natural genesis of ß-TCP/HAp phases in postnatal fishbones towards gold standard biocomposites for bone regeneration.

The search for gold-standard materials for bone regeneration is still a challenge in reconstruction surgery. The ratio between hydroxyapatite (HAp) and ß-tricalcium phosphate (ß-TCP) in biphasic calcium phosphate ceramics (BCPs) is one of the most important factors in osteoinduction promotion and controlled biodegradability, configurating what is currently considered as a possible gold standard material for bone substitution in reconstructive surgery. Exploring the natural genesis of the HAp and ß-TCP phases in fishbones during their postnatal growth, this study developed a biphasic bioceramic obtained from the calcination of Nile tilapia (Oreochromis niloticus) bones as a function of their ages. The natural genesis dynamics of the structural evolution of the ß-TCP and HAp phases were characterized by physicochemical methods, taking into account of the age of the fish and the material processing conditions. Thermal analysis (TGA / DTA) showed complete removal of the organic matter and transitions associated with the transformation of carbonated hydroxyapatite (CDHA) to HAp and ß-TCP phases. After calcination at 900 °C, the material was characterized by: X-ray diffraction (XRD) and refinement by the Rietveld method; Fourier Transform Infrared Spectroscopy with Attenuated Total Reflection (FTIR-ATR); Raman spectroscopy; Scanning Electron Microscopy (SEM) and Flame Atomic Absorption Spectroscopy (FAAS). The analysis allowed identification and quantitative estimate of the variations of the HAp and ß-TCP phases in the formation of the BCPs. The results showed that the decrease in ß-TCP against the increase in the HAp phases is symmetrical to the dynamics of the natural genesis of these phases, surprisingly maintaining the balanced phase proportion even when bones of young fishes were used. The microstructure analysis confirms the observed transformation. In addition, in vivo tests demonstrated the osteoinductive potential of BCP scaffolds implanted in an ectopic site, and their remarkable regenerative functionality, as bone graft, was demonstrated in alveolar bone after tooth extraction. MTT cytotoxicity assay for BCP samples for MC3T3-E1 pre-osteoblasts and L929 fibroblasts cells showed viability equal or higher than 100%. A logistic empirical model is presented to explain the three stages of HAp natural formation with fish age and it is also compared to the fish size evolution.

Integrative taxonomy: combining molecular and morphological characteristics to identify Lymnaea (Galba) cubensis, intermediate host of Fasciola hepatica.

Despite the epidemiological importance of the Lymnaeidae family regarding transmission of Fasciola hepatica, knowledge about the diversity and distribution of these molluscs and the role of each species in the expansion of fasciolosis remains sparse. Classical morphological (n=10) identification was performed in lymneids from Lagoa Santa, a municipality in the state of Minas Gerais, Brazil, along with molecular and phylogenetic analysis (n=05) based on the partial nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I gene (COI mtDNA) and ribosomal internal transcribed spacer II (ITS-2 rDNA). The shell morphology made it possible to distinguish the lymneids of Lagoa Santa from Pseudosuccinea columella. Differences found in the penile complex and prostate shape allowed this species to be distinguished from Galba truncatula. However, the homogeneity of reproductive tract characteristics among Lymnaea (Galba) cubensis, L. viator and L. neotropica confirmed that these characteristics show low taxonomic reliability for identifying cryptic species. Genetic divergence analysis for the COI mtDNA gene and ITS-2 region of rDNA revealed greater similarity to Lymnaea (Galba) cubensis. Thus, correct species differentiation is important for monitoring the epidemiological risk of fasciolosis in the state of Minas Gerais, where cases of the disease have increased over recent years.

The effects of resistance training performed in water on muscle strength in the elderly.

The purpose of the present study was to compare the effects of a program of resistance training in water-based exercises (RWE) with those of a program without resistance control in water-based exercises (WEs). Twenty-seven women (aged 60-74 years) were randomly assigned to the RWE group (n = 10), WE group (n = 10), or nontraining control (CON) group (n = 7). The RWE and WE groups trained classes with aerobic exercises and localized muscular resistance exercises for 50 minutes, twice a week for 12 weeks. For the RWE group, the program included 4 mesocycles of 3 weeks-respectively, 4 sets of 15 repetitions, 4 sets of 12 repetitions, 5 sets of 10 repetitions, and 5 sets of 8 repetitions-of shoulder horizontal flexion exercise at maximum speed, with the use of resistive equipment. For the WE group, the training was not periodized, and the resistance in the localized muscular exercises was not controlled. One repetition maximum (1RM) was measured on a pectoral fly machine at baseline and after the training period. The level of significance adopted was p ≤ 0.05. The results showed that the only significant increase in 1RM (10.89%, p < 0.001) occurred in the RWE group after training. In conclusion, these findings suggest that WEs with emphasis on resistance training in a periodized program can efficiently increase maximum strength in elderly women. Thus, it is suggested that the strategies used in WE programs be modified to offer suitable stimuli for the development of strength.

New approaches to studying morphological details of intramolluscan stages of Angiostrongylus vasorum.

Angiostrongylus vasorum is a pulmonary artery parasite of domestic and wild canid. On molluscs, intermediate host, first stage larvae (L1) are found after the first day of infection, in the 8th L2 and in the 30 th L3. It was evaluated L1, L2 and L3 recovered by Baermann technique from Achatina fulica infected with 1000 L1. Fifty larvae/stage were incubated with antibodies anti-ß-tubulin, anti-α-tubulin, anti- α-actin, anti-ß-actin and anti-collagen, and then with Alexa 633. Fifty larvae/stage were observed with picrosirius red and Oil Red O. It was also observed in the anterior region of L1 the beginning of the chitinous stems development, in the initial portion of the intestine and genital primordium. In L2 anterior region, the papillae, chitinous canes juxtaposed to the mouth and intestines bigger than L1. The L3 musculature is well defined, next to the chitinous stems, there are two round distally arranged from each other. It was observed the whole extension of the intestine genital primordium and intense cellularity in the L3 distal portion. With the picrosirius red the L1, L2 and L3 musculature could be observed, as the nerve ganglia on L3. Oil Red O revealed that L1, L2 and L3 store energy on lipid droplets.

Angiostrongylus minasensis n. sp.: new species found parasitizing coatis (Nasua nasua) in an urban protected area in Brazil.

Currently, there are 21 species of Angiostrongylus that parasitize the pulmonary or mesenteric arteries of wild and domestic rodents, felids, canids and human. Species of Angiostrongylus have cosmopolitan distribution covering tropical, subtropical and temperate regions. The procyonid Nasua nasua (coati) is a reservoir host for a wide variety of parasites that may be harmful to its populations or may contain etiological agents with zoonotic potential. In urban areas, coatis are usually found in close association with humans and domestic animals. We morphologically and molecularly characterized a new species of Angiostrongylus found in N. nasua in a protected area within Belo Horizonte, Brazil. The new species of Angiostrongylus differs from other species of the same genus in terms of the length and bifurcation level of the lateral and ventral rays, the length of spicules and female tail morphology. Molecular phylogenetic results based on the mitochondrial cytochrome c oxidase subunit 1 gene suggest that the newly identified species belongs to a genetic lineage that is separate from other species of Angiostrongylus. This new species was collected from the mesenteric arteries of N. nasua. It was named Angiostrongylus minasensis n. sp..

Nodular and sclerosing gastritis caused by Cylicospirura felineus in a puma (Puma concolor).

An adult male puma (Puma concolor), hit by a car in an urban area, died three days later despite the therapeutic support provided. At necropsy, multiple firm nodules were identified in the gastric mucosa. The nodules were coated by an intact mucosa with a central opening from which reddish and cylindrical nematodes protruded into the lumen. Twenty-seven nematodes were retrieved for morphological and morphometric evaluations. During histopathological examination of the gastric tissue, the adult nematodes appear in longitudinal and transverse sections, surrounded by thick bands of collagen, interspersed with mixed inflammatory infiltrates. The nematodes had an eosinophilic cuticle with caudal serrated projections (bulbar type), coelomyarian musculature, pseudocoelom, and females with uterus containing numerous larvated eggs, characteristics consistent with the Cylicospirura genus. Morphologically, female nematodes had six large tricuspid teeth in the oral cavity and the vulva had an opening anterior to the esophagus-intestinal junction. Male nematodes had five pairs of small papillae near the tip of the tail. These findings were consistent with Cylicospirura felineus. This parasite should be included in the differential diagnosis of nodular gastric wall lesions in wild felids.

Use of a candidate gene array to delineate gene expression patterns in cattle selected for resistance or susceptibility to intestinal nematodes.

In the present study, we use microarray technology to investigate the expression patterns of 381 genes with known association to host immune responses. Hybridization targets were derived from previously characterized bovine cDNAs. A total of 576 reporters (473 sequence-validated cDNAs and 77 controls) were spotted onto glass slides in two sets of four replicates. Two color, comparative hybridizations across both mesenteric lymph node (MLN) and small intestine mucosa (SIM) RNA samples were done between animals with previously demonstrated phenotypic differences based on natural exposure to gastrointestinal (GI) nematodes over a 6-month exposure period. A total of 138 significant hybridization differences were detected by mixed model analysis of variance. A subset of these significant differences was validated by quantitative, real-time RT-PCR to assay transcript levels for 18 genes. These results confirmed that in the SIM, susceptible animals showed significantly higher levels in the genes encoding IGHG1, CD3E, ACTB, IRF1, CCL5 and C3, while in the MLN of resistant animals, higher levels of expression were confirmed for PTPRC, CD1D and ITGA4. Combined, the results indicate that immune responses against GI nematode infections involve multiple response pathways. Higher levels of expression for IgE receptor, integrins, complement, monocyte/macrophage and tissue factors are related to resistance. In contrast, higher levels of expression for immunoglobulin chains and TCRs are related to susceptibility. Identification of these genes provides a framework to better understand the genetic variation underlying parasite resistance.

Competition for RISC binding predicts in vitro potency of siRNA.

Short interfering RNAs (siRNA) guide degradation of target RNA by the RNA-induced silencing complex (RISC). The use of siRNA in animals is limited partially due to the short half-life of siRNAs in tissues. Chemically modified siRNAs are necessary that maintain mRNA degradation activity, but are more stable to nucleases. In this study, we utilized alternating 2'-O-methyl and 2'-deoxy-2'-fluoro (OMe/F) chemically modified siRNA targeting PTEN and Eg5. OMe/F-modified siRNA consistently reduced mRNA and protein levels with equal or greater potency and efficacy than unmodified siRNA. We showed that modified siRNAs use the RISC mechanism and lead to cleavage of target mRNA at the same position as unmodified siRNA. We further demonstrated that siRNAs can compete with each other, where highly potent siRNAs can compete with less potent siRNAs, thus limiting the ability of siRNAs with lower potency to mediate mRNA degradation. In contrast, a siRNA with low potency cannot compete with a highly efficient siRNA. We established a correlation between siRNA potency and ability to compete with other siRNAs. Thus, siRNAs that are more potent inhibitors for mRNA destruction have the potential to out-compete less potent siRNAs indicating that the amount of a cellular component, perhaps RISC, limits siRNA activity.