Using several-fold higher concentrations of the test β-lactam antibiotic, compared to the probe, enhances the likelihood that the antibiotic will be the preferred substrate of the lactamase in the competition reaction in the assay. The reduced fluorescence indirectly reflects the ability of the β-lactamase to bind and cleave the tested antibiotic (large difference = antibiotic can be readily

bound and hence cleaved and inactivated). Notably, unlike growth based conventional AST methods, the end-point SCH727965 cell line of the β-LEAF assay is not bacterial viability or differences in growth pattern. The read-out of the assay is fluorescence, which reflects probe cleavage due to the enzymatic activity of the β-lactamase. Importantly, the β-LEAF assay is rapid compared to the conventional growth based AST methods (1 h versus 20–24 h for disk diffusion/MIC conventionally or ~8 h with automated instruments). The observation in Figure 2 of low to negligible fluorescence in β-LEAF + cefazolin reactions with all β-lactamase ‘positives’ (#1, #6, #18, #19, #20) suggests that cefazolin can be readily targeted and inactivated by the respective lactamases, and would be anticipated to be a less effective treatment option for these bacteria. An expectation of this assay is that the reduction in probe fluorescence in the presence of an antibiotic will be inversely proportional to its predicted activity

against the pathogen. If fluorescence is completely reduced in the presence of an antibiotic, then the respective antibiotic can be readily cleaved and inactivated by β-lactamase. Nepicastat chemical structure However, if despite the ‘saturating’ amount of antibiotic, some fluorescence selleck chemical increase reflecting probe cleavage is still observed (e.g. cefepime reactions in Figure 3), the lactamase may not be capable of effectively destroying the antibiotic, and the antibiotic predicted as likely to be active. In experiments with multiple antibiotics (Figure 3) a ratio of the cleavage rate of β-LEAF in presence of an antibiotic to the cleavage rate of β-LEAF alone, for each antibiotic tested, is shown in Table 4. For β-lactamase based resistance, the ratio of cleavage

rates closer to 1 (Table 4) would indicate greater β-lactam antibiotic efficacy. With more rigorous testing Sclareol from multiple data sets on a large number of isolates, cut-offs could be set up to develop the ratios as a ‘β-lactamase-based antibiotic activity/susceptibility index’ within specific limits. We recognize that there are a wide variety of lactamases, and note that with appropriate kinetic analysis (such as building on our previous study [50]), the approach presented here has the potential of characterizing the different lactamases. The motivation for the choice of antibiotics used in this initial study was to test three different generations of cephalosporin antibiotics. Cephalosporins are a standard treatment for skin and soft-tissue infections [58, 59].

F. (2002). A self-replicating ligase ribozyme. Proc. Natl. Acad. Sci. USA

99:12733–12740. Robertson, M. P. and Ellington, A. D. (1999). In vitro selection of an allosteric ribozyme IWR1 that transduces analytes into amplicons. Nature Biotechnol. 17:62– 66. Rogers, J. and Joyce, G. F. (2001). The effect of cytidine on the structure and function of an RNA ligase ribozyme. RNA 7:395–404. E-mail: gjoyce@scripps.​edu Cosmochemical Evolution and the Origins of Life: A Tribute to Joan Oró Sandra Pizzarello Arizona State University, Tempe AZ 85287–1604 USA Joan (John) Oró was an enthusiastic and eclectic exobiologist who, since the early days of the learn more discipline, promoted the idea of cosmochemical evolution as a possible precursor to terrestrial life (Oró, 1961). The idea also made him a pioneer in meteoritic studies, as he recognized the importance of natural sample analyses towards the understanding and modeling of life’s origins. This lecture in his honor will tell of new types of meteorites and the advances that their analyses have brought to our knowledge of prebiotic extraterrestrial

chemistry. Carbonaceous meteorites provide a detailed record of the organic materials that can be synthesized in abiotic environments. These have been shown to be complex and to have structures as varied as kerogen-like macromolecules and simpler soluble compounds, e.g., amino acids and hydrocarbons (Pizzarello et al., 2006). Meteorite organics display an overall molecular and isotopic diversity that points to synthetic pathways in a variety of Temsirolimus chemical regimes, such as exothermic reactions in the cold, hydrogen fractionating interstellar gas phase and aqueous reactions in asteroidal parent bodies. Within this diversity, some meteoritic compounds have been found to be identical to biomolecules, with some of the amino acids displaying the biochemical trait of chiral asymmetry. This, in turn, has suggested that their delivery to the early Earth might have contributed to terrestrial molecular evolution (Pizzarello, 2006). Yet, so far, the study of meteorites has been hindered by the fact that the carbonaceous types are few

in recorded falls (only 18 in the last two centuries), are often lost or irreparably altered after their fall and Cytidine deaminase that their soluble organic content degrades with terrestrial exposure (Cronin et al., 1980). This fate may be spared to the stones recovered in Antarctica, where in-falling meteorites are quickly covered by snow, buried within the ice and resurface only when the flowing ice sheets end-up against the obstacle of a mountain. Owing to this unique shelter of the glaciers, American and Japanese scientific expeditions have found here a large number of carbonaceous meteorites, some of which are unspoiled. We will report on the organic composition of two pristine Antarctic meteorites belonging to the Renazzo-type group.

Involvement of the heat-labile serum factor suggests the potential role of the complement for defensin expression. The possible link between the proteins of the complement system and defensin expression may be anticipated since the interaction between the defensins and proteins of the complement system was demonstrated. It was found that HBD2 inhibits the classical pathway of the complement system [37]. Moreover, the interrelationship between the respiratory tract and the complement system was studied in an animal model [38]. The origin of complement proteins present CHIR-99021 solubility dmso in the lining fluid of the respiratory tract

is thought to result mainly from plasma that exudes into the bronchoalveolar space. However, it was shown that human bronchial epithelial cells generate complement protein C3: the modulation

of its expression by proinflammatory cytokines might be an additional regulatory mechanism of local airway defence during infection [39]. Furthermore, the kinetics of the expression of human beta defensins, hBD2 and hBD9, by airway epithelial cells exposed to the deferent morphotypes of A. fumigatus was analysed. Analysis of the kinetics of hBD2 and hBD9 defensin expression by cells exposed to A. fumigatus showed the prompt inducible expression of hBD9, following by delayed hBD2 expression. This could allow us to hypothesize that the host immune system may react against A. fumigatus by using the cascade of newly synthesized defensins that probably possess the different functions. OICR-9429 cell line However, this hypothesis would require further investigation at the protein level. Our Cell Penetrating Peptide data are in agreement with the analysis of kinetics of hBD2 expression by A549 cells exposed to Mycobacterium tuberculosis; infection of A549 cells resulted in hBD2 gene expression as early as 6 hours postinfection, while maximum expression was detected at 18 and 24 hours postinfection [35]. Several lines of evidence eliminated the possibility that observed inducible defensin expression was related to endotoxin contamination of A. fumigatus organisms. First, the addition of Polymixin B (known

for its endotoxin-neutralising activity) to the cells prior to exposure to A. fumigatus organisms did not have any effect on defensin expression. Second, rigorous measures were undertaken to keep endotoxin out of the experimental system, including washing of A. fumigatus organisms with the solution containing Polymixin B during preparation, utilisation of endotoxin-free plasticware and solutions in experiments, and washing of fungal organisms in endotoxin-free PBS prior to use. The expression of hBD2 and hBD9 was found to be higher in A549, 16HBE and primary culture HNT cells exposed to SC compared to RC or HF, as shown by quantitative PCR. During asexual growth, the morphological form of A. fumigatus changes from resting to swollen conidia, which then form germ tubes that continue growing in hyphal form. These transformations are accompanied by the modification of BIBF 1120 surface structures.

0398). Table 2 Correlation between gene expression and GEM efficacy in patients with pancreatic cancer receiving GEM monotherapy.     GEM efficacy   Gene Expression* https://www.selleckchem.com/products/a-1210477.html   Effective§ Non-effective P ¶-value hENT1 High 4 9 >0.9999   Low 8 14   hENT2 High 6 9 0.5374   Low 6 14   dCK High 8 7 0.0398   Low 4 16   DCD High 3 9 0.4765   Low 9 14   CDA High 4 9 >0.9999   Low 8 14   5′-NT High 4 12 0.2882   Low 8 11   RRM1 High 4 8 >0.9999   Low 8 15   RRM2 High 4 8 >0.9999   Low 8 15   GEM, gemcitabine *Gene expression was determined as high or low based on mean values of 35 EUS-FNA samples. §Effective, partial response by imaging study

or stable disease by imaging study with 50% or more decrease in tumor markers compared to pretreatment value ¶ P, examined by chi-squared test (Fisher’s exact test) Discussion EUS-FNA is widely used as a cytological and histological diagnostic method for pancreatic cancer [8, 11].

However, there have been few reports on gene analysis of pancreatic cancer using EUS-FNA samples [7, 8, 12]. In contrast, a number of selleck chemicals llc studies have demonstrated the feasibility of DNA microarray analysis using samples obtained by FNA in other malignancies, such as breast cancer and lung cancer [13–15]. At least 10 μg of total RNA is required for DNA microarray analysis [10]. Due to the low volume of biopsy specimens obtained by EUS-FNA, it is Alvocidib chemical structure typically impossible to perform DNA microarray analysis using the raw RNA extracted from these samples. However, a high-fidelity RNA amplification protocol has recently been established [10, 16] that allows analysis of gene expression profiles using small volumes RNA, such as those obtained by EUS-FNA. In our series, only 0.1 – 3.0 μg of total RNA was extracted from EUS-FNA biopsy samples. The objective response rate of GEM monotherapy for pancreatic cancer has been reported to be 5–12% [1, 17, 18]. In this study, PR was observed in 5 of 35 (14%) patients treated with GEM monotherapy, which corresponds

with the response rates reported previously. The number of patients in the GEM-effective group was too buy MG-132 small to evaluate for correlations between GEM efficacy and mRNA expression. Therefore, SD patients with a 50% or more decrease in abnormal serum levels of tumor markers compared to baseline were included in the GEM-effective group. CA 19-9 has been shown to be correlated with clinical efficacy of GEM in pancreatic cancer [19]. In this study, the GEM-effective group had a significantly better prognosis than the non-effective group, indicating that the grouping based on GEM efficacy was appropriate. GEM is transported into the cell largely via hENT1 and partly via hENT2 [4].

The population of our registry CFTRinh-172 in vivo was relatively old (mean age approximately 65 years). The age of the study population may have meant that

there was a higher proportion of patients with isolated systolic hypertension (ISH) than would have been seen for a study with a younger population. However, baseline BP measurements were averaged, so it was not possible to determine the proportion of patients with ISH. Patients with ISH have marked arterial stiffening, which makes BP control more difficult. In light of the possibility that a significant proportion of patients in our study could have had ISH, the BP-lowering and BP control rates observed are even more impressive. Our results are comparable to those seen in a study in elderly patients (age 60–85 years), in which treatment with the combination of lercanidipine 10 mg plus https://www.selleckchem.com/products/prt062607-p505-15-hcl.html enalapril 20 mg for 4 weeks was associated with a reduction in SBP of 16.9 mmHg compared with baseline, and a BP control rate of 45 % [20]. In this study, the BP-reducing effect of lercanidipine/enalapril was greater in patients receiving lercanidipine/enalapril alone compared with patients receiving the FDC with other antihypertensive drugs. However, at the end of the study period, the mean BP values and BP control rates Dasatinib in

both patient groups were similar. This can best be explained by the fact that the magnitude of the therapeutic benefit is generally correlated with baseline BP values [22]. As the patients who received lercanidipine/enalapril alone had significantly greater baseline BP values and lower BP control rates than those who received lercanidipine/enalapril with other antihypertensive drugs, the greater magnitude of improvement

at the end of the study in patients who received lercanidipine/enalapril alone was expected. The introduction of this FDC, in addition to the noted efficacy, did not significantly increase the number of drugs required to ADP ribosylation factor achieve BP control. These results may be particularly interesting from an economic perspective, as a reduction in the number of concomitant medications has the potential to produce cost savings, particularly for a high-prevalence disease such as hypertension. The primary limitation of this study was that it was an open-label pharmaco-epidemiological registry, with all the inherent limitations and advantages of such a design. Other limitations were the relatively small number of patients and the short follow-up duration. The size of the study was necessarily limited by the number of patients presenting to CCG members’ clinics during the study period for whom the lercanidipine/enalapril (10/20 mg) FDC was considered the most appropriate treatment.

New Phytol 2005,165(1):215–226.Pevonedistat clinical trial PubMedCrossRef 70. Baier R, Schiene K, Kohring B, Flaschel E, Niehaus K: Alfalfa and tobacco cells react differently to chitin oligosaccharides and Sinorhizobium meliloti nodulation factors. Planta 1999,210(1):157–164.PubMedCrossRef 71. Felix G, Duran JD, Volko S, Boller T: Plants have a sensitive perception system for the most conserved domain

of bacterial flagellin. Plant J 1999,18(3):265–276.PubMedCrossRef 72. Gomez-Gomez L, Boller T: Flagellin perception: a paradigm for innate immunity. Trends Plant Sci 2002,7(6):251–256.PubMedCrossRef 73. Nürnberger T, Wirtz W, Nennstiel D, Hahlbrock K, Jabs T, Zimmermann S, Scheel D: Signal perception and intracellular signal transduction in plant pathogen defense. J Recept Signal Transduct Res 1997,17(1–3):127–136.PubMed 74. Rouet-Mayer selleck inhibitor M-A, Mathieu Y, Cazale A-C, Guern J, Lauriere C: Extracellular alkalinization and

oxidative burst induced by fungal lyase in tobacco cells are not due to the perception of oligogalacturonide fragments. Plant Physiol Biochem 1997,35(4):321–330. 75. Hardy MR, Townsend RR: Separation of positional isomers of oligosaccharides and glycopeptides by high-performance anion-exchange chromatography with pulsed amperometric detection. Proc Natl Acad Sci USA 1988,85(10):3289–3293.PubMedCrossRef learn more 76. Moerschbacher BM, RVX-208 Mierau M, Graessner B, Noll U, Mort AJ: Small oligomers of galacturonic

acid are endogenous suppressors of disease resistance reactions in wheat leaves. J Exp Bot 1999,50(334):605–612.CrossRef 77. Svalheim O, Robertsen B: Elicitation of H2O2 production in cucumber hypocotyl segments by oligo-1,4-alpha-D-galacturonides and an oligo-beta-glucan preparation from cell walls of Phythophthora megasperma F Sp glycinea. Physiol Plantarum 1993,88(4):675–681.CrossRef 78. Ryan CA: Oligosaccharides as recognition signals for the expression of defensive genes in plants. Biochemistry 1988,27(25):8879–8883.CrossRef 79. Norman C, Vidal S, Palva ET: Oligogalacturonide-mediated induction of a gene involved in jasmonic acid synthesis in response to the cell-wall-degrading enzymes of the plant pathogen Erwinia carotovora . Mol Plant Microbe Interact 1999,12(7):640–644.PubMedCrossRef 80. Stamp N: Out of the quagmire of plant defense hypotheses. Q Rev Biol 2003,78(1):23–55.PubMedCrossRef 81. Büttner D, Bonas U: Common infection strategies of plant and animal pathogenic bacteria. Curr Opin Plant Biol 2003,6(4):312–319.PubMedCrossRef 82. Kunze G, Zipfel C, Robatzek S, Niehaus K, Boller T, Felix G: The N terminus of bacterial elongation factor Tu elicits innate immunity in Arabidopsis plants . Plant Cell 2004,16(12):3496–3507.PubMedCrossRef 83.

crenulatum a high negative osmotic potential of—2.09 MPa has been determined by incipient plasmolysis (equivalent to an osmolarity of 961 mOsm kg−1), which substantially contributes to its water-holding capacities (Kaplan et al. 2012). The ultrastructural appearance upon treatment with

sorbitol leads to a condensed cytoplasm similar to that in the desiccation experiments. The cell walls, however, do not shrink in the hyperosmotic solutions, but remain connected with the plasmolysed cytoplasm via Hechtian strands (Kaplan et al. 2012). The osmotic potential of semi-terrestrial Zygnema is less negative in younger developmental stages, but increases upon the formation of akinetes (Kaplan et al. 2013). In nature, these akinete stages are found only in late summer (e.g., Holzinger et al. 2009), thus providing the capacity to AMN-107 price survive desiccation. Fig. 5 Transmission 4SC-202 cost electron micrographs of Klebsormidium crenulatum (SAG 2415), a desiccated at 95 % air relative humidity for 4 days, b desiccated at 5 % air relative humidity for 4 days, c, d plasmolysed with 1,000 mM sorbitol for 3 h. The general appearance of the cytoplasm is similarly dense JQ-EZ-05 solubility dmso regardless of the different treatments, except that in desiccated samples the cross walls appear undulated (a). oCW outer cell wall,

cCW cross cell wall, Chl chloroplast, M mitochondrion, N nucleus, P peroxisome, S starch, V vacuole. Bars 1 μm. a, b reprinted from Holzinger et al. (2011) with permission of the Phycological Society of America; c, d reprinted from Kaplan et al. (2012) with permission of Springer Science and Business Media Protective

strategies against desiccation in alpine biological soil crust algae Eukaryotic algae in BSCs have evolved avoidance and protection strategies to maintain integrity under unfavorable water-potential conditions. So far, little is understood on the community level, but self-protection may be important, as the vertically Acyl CoA dehydrogenase lower-positioned organisms of a soil crust may not even be exposed to water stress due to the water-holding capacities of the organisms on top and in the crust matrix. A biochemical protection strategy is the production of osmotically active carbohydrates such as polyols, generated particularly by green algae from the Trebouxiophyceae (e.g., Gustavs et al. 2010). However, these compounds are lacking in reasonable concentrations in Klebsormidiophyceae, which are the major component organisms in alpine BSCs (Kaplan et al. 2012). An organized ‘shutdown’ of PSII occurs during desiccation in BSC algae (Karsten et al. 2010; Karsten and Holzinger 2012). Dynamic photoinhibition has recently been confirmed for several species of desert and aquatic green algae (Lunch et al. 2013). Although photoprotective mechanisms in those green algae that have been investigated are similar, the mechanisms exhibit lineage-specific differences.

In addition, cross-links to improve stability of the implanted system are not available for minimal-invasive implantation. Therefore a conventional open approach should be performed to allow for an uncompromised reduction of the spinal injury, especially in regard to eventual secondary anterior column surgery (see Figure 4). On the other hand, if sufficient reduction during posture and following traction or cautious manipulation of the patient is achieved, one should keep in mind percutaneous fixation in those rare cases [24]. Figure 4 Conventional open reduction and instrumentation

with secondary anterior surgery in a polytraumatized NVP-HSP990 patient with compression fracture of T12 and complete burst fracture of L1. This case features a 39 year old male patient following a fall from height (ISS = 41). The patient was unconscious at the site of the injury and transferred after tracheal intubation to the trauma centre. Following primary survey and whole-body CT-Scan, severe Selleckchem NU7026 traumatic brain injury with epidural hematoma, retroperitoneal bleeding with bilateral lung contusions and instable spine injuries from a complete burst fracture of L1 with substantial spinal canal

compromise (type A3.3) and adjacent compression fracture of T12 (type A1.2) were revealed (images VX-661 mouse A-D). The patient was positioned prone and simultaneous surgery was performed for evacuation of epidural hematoma and stabilization of the spine. Posterior fusion using a conventional approach was performed to achieve optimized reduction of the posterior wall fragment and strongest stabilization using a cross-link and bone graft (image E). Following uneventful recovery from intracranial injuries, the patient was operated anterior using an expandable cage on day 10 post trauma (images F-G). Removal

of the internal fixator after 14 months released cranial motion segment T11-T12 and showed sufficient bisegmental oxyclozanide anterior fusion (images H-I). (Adopted from Heyde CE, Stahel PF, Ertel W. “”Was gibt es Neues in der Unfallchirurgie”" in: Meßmer, Jähne, Neuhaus: Was gibt es Neues in der Chirurgie? Ecomed Medizin 2005). What to do with neurologic deficit in the first operative phase? Considering spinal cord injury, a vast array of research efforts have been undertaken for we kindly refer the reader to the current literature and reviews. The consensus has been established, that a mechanical impact to the spinal cord initiates and entertains secondary injury events, that exacerbate the spinal cord injury [43, 97], as it is also evident for traumatic brain injury [41, 42]. As a consequence, spinal cord decompression has to be performed even in the polytraumatized patient [30] and this as quick as possible, since decompression between 24 h and 72 h is shown to be too late to prevent substantial neurologic deficits [98–102].

4H2O: the first occurrence of a condensed phosphate as a mineral. Am Min 73:168–171 Russell MJ, Hall AJ (1997) The emergence of life from iron monosulphide bubbles at a submarine hydrothermal redox and pH front. J Geol Soc London 154:377–402PubMedCrossRef Sales BC, Chakoumakos BC, Boatner LA, Ramey JO (1993) Structural properties of the amorphous phases produced by heating crystalline MgHPO 4 . 3H2O. J Non-Cryst Solids 159:121–139CrossRef Schoonen M, Smirnov A, Cohn C (2004) A perspective on the role of minerals in prebiotic synthesis. Ambio 33:539–551PubMed Schwartz AW (1971) Phosphate: solubilization and

activation on the early Earth. In: Buvet R, Ponnamperuma C (eds) Chemical Evolution and the Origin of Life, North-Holland, Amsterdam, pp 100–108 Schwartz AW (2006) Phosphorus in prebiotic chemistry. Phil Trans R Soc B 361:1743–1749PubMedCrossRef Seel F, Klos

KP, Schuh J (1985) Hydrothermale Kondensation von Magnesium-hydrogenphosphaten GDC-0449 ic50 zu Magnesiumdiphosphaten. Naturwissenschaften 72:658CrossRef Seel F, Klos KP, Rechtenwald D, Schuh J (1986) Non-enzymatic formation of condensed phosphates under prebiotic conditions. Z Naturforsch B 41B:815–824 Serrano A, Pérez-Castiñeira JR, this website Baltscheffsky M, Baltscheffsky H (2007) H+−PPases: yesterday, today and tomorrow. IUBMB Life 59(2):76–83PubMedCrossRef Seyfried WE Jr, Foustoukos DI, Fu Q (2007) Redox evolution and mass transfer during serpentinization: an experimental and theoretical study at 200°C, 500 bar with implications for ultramafic-hosted hydrothermal systems at Mid-Ocean Ridges. Geochim Cosmochim Acta 71:3872–3886CrossRef Skulachev VP LEE011 concentration (1996) Evolution of convertible energy currencies of the living cell: from ATP toΔμH + and ΔμNa+. In: Baltscheffsky H (ed) Origin and evolution of biological energy conversion. VCH, New York, pp 11–41 Staudigel H, Hart SR, Richardson SH (1981) Alteration of the oceanic crust: processes and timing.

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66. Guimarães N, Azevedo NF, Figueiredo C, Keevil CW, Vieira MJ: Development and application of a novel peptide nucleic acid probe for the specific detection of Helicobacter pylori in gastric biopsy specimens. J Clin Microbiol 2007,45(9):3089–3094.PubMedCrossRef Authors’ contributions MSG participated in the experimental design, carried out all experimental

work and drafted the manuscript. NFA, SAW, MJV and CWK participated in the design of the study and helped to draft the manuscript. All authors have read and approved the final manuscript.”
“Background Infectious diseases have devastating ecological and economical impacts on fish, amphibian and reptile populations worldwide (reviewed in [1]). Despite those effects, the precise TSA HDAC chemical structure pathogenesis of infectious diseases of ectotherm vertebrates and the interaction with the immune system of their respective hosts are mostly poorly understood. Recently, Navitoclax order marked progress has been made in the characterization of the immune system of lower vertebrates. This has been facilitated by concentrated focus on the cloning of pathogen-induced genes and by accumulating sequence data from genome and expressed sequence tag (EST) projects. Similarly, increased information about the genomes of pathogens of lower vertebrates is becoming available. However, there are still large gaps in our knowledge, Phospholipase D1 especially concerning the interaction of ectothermic pathogens with the host immune system. Ranaviruses, which constitute a genus within the family Iridoviridae, are important pathogens of ectotherms

and have been associated with massive die-offs of both wild and farmed populations of fish, frogs and salamanders in diverse areas of the world [2–5]. Ranaviruses are double-stranded DNA viruses with genomes ranging from 105 to 140 kb. Currently the genomes of seven ranaviruses have been sequenced: Ambystoma tigrinum virus (ATV, accession no. NC_005832[6]); Frog virus 3 (FV3, accession no. NC_005946[7]); Tiger frog virus (TFV, accession no. AF389451 [8]); Grouper iridovirus (GIV,accession no. AY666015 [9]; Singapore grouper iridovirus (SGIV, accession no. NC_006549[10]); Soft-shelled turtle iridovirus (STIV, accession no. EU627010 [11]); and Epizootic hematopoietic necrosis virus (EHNV, accession no. FJ433873 [12]). Phylogenetic analysis showed the existence of two major clades among ranaviruses, one that included GIV and SGIV, and another comprised of ATV, EHNV, FV3, STIV and TFV. Interestingly, the latter clade could be further subdivided with ATV and EHNV in one subclade, and FV3, STIV and TFV in the other.