3). To investigate whether the uptake of SpHtp1 can be caused by physical disruption of the membranes by Saprolegnia, a His-tagged SpHtp1 fusion protein without the putative signal peptide, SpHtp124-198-His, was synthesized in E. coli, purified and characterized (Fig. S6). Treatment with the final bleed SpHtp1 antibody in combination with the secondary antibody Fluor 488 showed no fluorescence in or on RTG-2 cells.

When the RTG-2 cells were treated with SpHtp124-198-His, no fluorescence was detected when the preimmune antiserum was used in combination KU-60019 with the secondary antibody Fluor 488, also showing that the treatment of SpHtp1-His did not affect the fish cells (Fig. 4). However, SpHtp124-198-His and final bleed SpHtp1 antibody-treated RTG-2

cells showed SpHtp124-198-His localization on the surface of the fish cells and also inside the fish cells, surrounding the nucleus (Fig. Selleckchem DAPT 4). Furthermore, when the fish cells were incubated with SpHtp124-198-His and only labelled with the primary or the secondary antibody, no fluorescence was observed inside or outside the cells. Identical results were observed when an anti-His antibody was used for the immunolocalization studies (Fig. S7). Setting up a model infection system without having to sacrifice animals has many obvious advantages as it helps to fulfil the ultimate goal of the three Rs, whereby the aim is to reduce, refine and replace animals in experimental research. Here, we have shown that the trout RTG-2 cell line represents an excellent in vitro system for studying the very early interactions between

fish cells and S. parasitica, and that it can also be used to study the molecular mechanism of infection. Analysis of ESTs from zoospores and germinated cysts resulted in the identification of a putative RxLR effector protein, demonstrating that these types of proteins are possibly not only present in plant pathogenic oomycetes but also in animal pathogenic Florfenicol oomycetes. SpHtp1 is expressed in the preinfection and the very early infection stages of S. parasitica, as are many RxLR effector genes from P. sojae and P. infestans (Whisson et al., 2007; Dong et al., 2009). Analysis of the protein sequence revealed that SpHtp1 lacks the ‘so-called’ EER motif, which is found closely behind the RxLR motif in about 500 putative P. infestans RxLR effector proteins (Whisson et al., 2007) (Fig. 1a and b). The EER motif in the PiAvr3a and PsAvr1b proteins seems to be required for effector translocation of P. infestans and P. sojae, respectively (Whisson et al., 2007; Dou et al., 2008a). However, another intracellularly recognized RxLR effector protein, Atr13 of Hyaloperonospora arabidopsidis, lacks the EER motif (Allen et al., 2004), suggesting that the presence of an EER motif is not always essential for the translocation of every RxLR effector into host cells, or for inducing a hypersensitive response.

7% (95% CI 19.9–37.5%). Our study reflects AZD4547 cell line the need to monitor the evolution of resistance on a regular basis and trends of transmitted resistance. The initiation of highly active antiretroviral therapy (HAART) in developing countries where HIV-1 non-B subtypes circulate has been associated with good clinical outcomes when combined with appropriate clinical follow-up [1]. However, as HAART is scaled up, it is essential to monitor the emergence

of primary resistance, as this may impact on the success of an already limited choice of first-line therapies in resource-limited settings [2]. Furthermore, studies have shown that polymorphisms in non-B subtype genomes can lead to different pathways to drug resistance from those found in subtype B HIV-1 [3–5]. We have studied the rate of primary resistance in Mali, a resource-limited country in West Africa. With a population of 11 million inhabitants, Mali has an estimated HIV prevalence of 1.3%, representing 146 000 persons infected with HIV [6]. The first antiretroviral drugs became available in 1997, followed by roll-out of HAART through a national treatment programme in 2004 with stavudine, lamivudine and nevirapine recommended as first-line treatment [7]. A study in 2006 estimated that the overall prevalence of primary resistance in Mali was 11.5% [7]. In the context of the scale-up of HAART, we therefore decided to evaluate the evolution of primary

resistance in this country. A total of 101 antiretroviral-naïve HIV-infected individuals from Mali were prospectively enrolled in this study. Primary resistance was evaluated during the period from July 2007 to October Daporinad Liothyronine Sodium 2008. Individuals were recruited from three different sites in Bamako, Mali’s capital: 42 patients were recruited from the Centre d’Écoute de Soins, d’Animation et de Conseil (CESAC), which offers diagnostic services and care to HIV-infected individuals of rural and urban origin, 43 from the Gabriel Touré Hospital (HGT), and

16 from the Point G Hospital (HPG). HGT and HPG are the two largest hospitals in Mali. Although their patient populations are mainly urban, they are reference centres and see referrals from the entire country. Samples obtained from the patients were stored at −80 °C until they were sent on dry ice for genotyping at the retrovirology laboratory at the Centre Hospitalier de l’Université de Montréal (CHUM), Montréal, Canada. This study was approved by the ethics committees of Mali and CHUM research centre. Viral RNA was extracted using the viral QIAamp Spin Mini Kit (Qiagen, Mississauga, Ontario, Canada) and according to the protocol provided by Virco (Mechelem, Belgium). It was then amplified using Superscript III HIFI (Invitrogen, Carlsbad, CA, USA) with primers 5′out and 3′RT (Virco) covering the protease and reverse transcriptase (RT-PR) genes. For the nested PCR, we used the expand HF PCR (Roche Applied Science, Quebec, Canada) as the PCR enzyme and primers 5′IN and 3′IN (Virco).

7% (95% CI 19.9–37.5%). Our study reflects GSK126 the need to monitor the evolution of resistance on a regular basis and trends of transmitted resistance. The initiation of highly active antiretroviral therapy (HAART) in developing countries where HIV-1 non-B subtypes circulate has been associated with good clinical outcomes when combined with appropriate clinical follow-up [1]. However, as HAART is scaled up, it is essential to monitor the emergence

of primary resistance, as this may impact on the success of an already limited choice of first-line therapies in resource-limited settings [2]. Furthermore, studies have shown that polymorphisms in non-B subtype genomes can lead to different pathways to drug resistance from those found in subtype B HIV-1 [3–5]. We have studied the rate of primary resistance in Mali, a resource-limited country in West Africa. With a population of 11 million inhabitants, Mali has an estimated HIV prevalence of 1.3%, representing 146 000 persons infected with HIV [6]. The first antiretroviral drugs became available in 1997, followed by roll-out of HAART through a national treatment programme in 2004 with stavudine, lamivudine and nevirapine recommended as first-line treatment [7]. A study in 2006 estimated that the overall prevalence of primary resistance in Mali was 11.5% [7]. In the context of the scale-up of HAART, we therefore decided to evaluate the evolution of primary

resistance in this country. A total of 101 antiretroviral-naïve HIV-infected individuals from Mali were prospectively enrolled in this study. Primary resistance was evaluated during the period from July 2007 to October DAPT chemical structure selleck compound 2008. Individuals were recruited from three different sites in Bamako, Mali’s capital: 42 patients were recruited from the Centre d’Écoute de Soins, d’Animation et de Conseil (CESAC), which offers diagnostic services and care to HIV-infected individuals of rural and urban origin, 43 from the Gabriel Touré Hospital (HGT), and

16 from the Point G Hospital (HPG). HGT and HPG are the two largest hospitals in Mali. Although their patient populations are mainly urban, they are reference centres and see referrals from the entire country. Samples obtained from the patients were stored at −80 °C until they were sent on dry ice for genotyping at the retrovirology laboratory at the Centre Hospitalier de l’Université de Montréal (CHUM), Montréal, Canada. This study was approved by the ethics committees of Mali and CHUM research centre. Viral RNA was extracted using the viral QIAamp Spin Mini Kit (Qiagen, Mississauga, Ontario, Canada) and according to the protocol provided by Virco (Mechelem, Belgium). It was then amplified using Superscript III HIFI (Invitrogen, Carlsbad, CA, USA) with primers 5′out and 3′RT (Virco) covering the protease and reverse transcriptase (RT-PR) genes. For the nested PCR, we used the expand HF PCR (Roche Applied Science, Quebec, Canada) as the PCR enzyme and primers 5′IN and 3′IN (Virco).

The groups were subdivided and immersed in: A (saliva), B (coffee), and C (wine). The baseline color was evaluated by spectrophotometer and repeated after 4 and 8 weeks, and after polishing, at the end of 8 weeks. The variation in color (∆E) and lightness (∆L) was analyzed by anova (two-way) and Tukey tests, and Friedman and Kruskal–Wallis tests, respectively. All specimens underwent color and lightness change, irrespective of immersion medium. In coffee, G2 presented the lowest mean ∆E (P < 0.05), compared with the other groups. In saliva,

G3 presented the highest mean ∆E, and G2 and G4 lower ∆E means. Lesions infiltrated with Icon® underwent greater color change when compared with remineralized lesions, which may represent an esthetic disadvantage for the first-mentioned treatment. “
“Early childhood caries (ECC) describes selleckchem dental caries affecting children aged 0–71 months.

Current research suggests ECC has important aetiological bases during the first year of life. Gaps in knowledge about disease progression prevent the effective and early identification of ‘at risk’ children. To conduct a systematic review of research studies focusing on (a) acquisition and colonization of oral bacteria and ECC and (b) risk and/or protective factors in infants aged 0–12 months. 17-AAG solubility dmso Ovid Medline and Embase databases (1996–2011) were searched for RCT, longitudinal, cross-sectional and qualitative studies. Two investigators undertook a quality assessment for risk of bias.

Inclusion criteria were met for (a) by four papers and for (b) by 13 papers; five papers were rated medium or high quality. Bacterial acquisition/colonization and modifying factor interrelationships were identified, but their role in the caries process was not clarified. Key risk indicators were infant feeding practices (nine papers), maternal circumstances and oral health (6) and infant-related oral health behaviours (4). This review confirmed that factors occurring during the first year of life affect ECC experience. Despite heterogeneity, findings indicated maternal factors influence bacterial acquisition, whereas colonization was mediated by oral health behaviours and practices and feeding habits. “
“Salivary osmolality reflects the hydration status of individuals with cerebral palsy (CP) necessary for an adequate unstimulated salivary Urease flow rate. To investigate whether salivary osmolality could serve as a potential indicator of caries risk in children with spastic CP by displaying a stronger association with caries occurrence than salivary flow rate. The convenience sample consisted of 65 children with CP aged 6–13 years old. Unstimulated whole saliva was collected using cotton roll, and salivary osmolality was measured using a freezing point depression osmometer. The children’s oral motor performance was evaluated during the feeding process using the Oral Motor Assessment Scale.

Most Dutch travel health nurses aspire to prescribe and feel competent to prescribe. Further education is required before implementing nurse prescribing in travel medicine. As this is the first study to focus on nurse prescribing in travel medicine, evaluation of travel nurse prescribing is strongly recommended and should start directly after the new responsibilities

are implemented. The authors declare that they have no competing interests. “
“We sought to evaluate and provide better itinerary-specific care to precounseled travelers and to assess diseases occurring while traveling abroad by surveying a community population. An additional quality improvement initiative was to expand our post-travel survey to

be a more valuable tool in Fulvestrant in vitro gathering high-quality quantitative ABT-737 data. From de-identified data collected via post-travel surveys, we identified a cohort of 525 patients for a retrospective observational analysis. We analyzed illness encountered while abroad, medication use, and whether a physician was consulted. We also examined itinerary variables, including continents and countries visited. The 525 post-travel surveys collected showed that the majority of respondents traveled to Asia (31%) or Africa (30%). The mean number of travel days was 21.3 (median, 14). Univariate analysis demonstrated a statistically significant increase of risk for general illness when comparing travel duration of less than 14 days to greater than 14 days (11.3% vs 27.7%, p < 0.001). Duration of travel was also significant with regard to development of traveler's diarrhea (TD) (p = 0.0015). Destination of travel and development of traveler's diarrhea trended toward significance. Serious illness requiring a physician visit was infrequent, as were vaccine-related complications.

Despite pre-travel counseling, traveler’s diarrhea was the most common illness in our cohort; expanded prevention strategies will be necessary to lower the impact that diarrheal illness has on generally healthy travelers. Overall rates of illness did not vary by destination; however, there was a strong association between duration of travel and likelihood of illness. To further identify specific variables contributing to travel-related disease, including Mannose-binding protein-associated serine protease patient co-morbidities, reason for travel, and accommodations, the post-travel survey has been modified and expanded. A limitation of this study was the low survey response rate (18%); to improve the return rate, we plan to implement supplemental modalities including email and a web-based database. In 2011, as reported by the World Tourism Organization, 980 million travelers crossed an international border. This number contrasts with 675 million international departures of only 10 years ago.[1] Following this explosive increase in international travel, the practice of travel medicine continues to grow.

Communication requires adaptability throughout the life of an individual, especially in species for which breeding periods (when intersexual signaling prevails) are interspersed with more ‘social’ (non-sexual) periods when intrasexual bonding prevails. In songbirds, structure or frequency of songs or song elements may convey different information depending on the season. This is the case in the European starling, where some song structures characterize social bonds between

females while other song structures are more characteristic of male courtship. We hypothesized that the female perceptual system may have adapted to these changes in song structure and function according to season, and we tested click here for potential seasonal brain plasticity. Electrophysiological recordings from adult female starlings during playback of song elements with different functions showed clear seasonal (breeding/non-breeding) changes in neuronal http://www.selleckchem.com/products/ganetespib-sta-9090.html responses in the primary auditory area. The proportion of responsive sites was higher in response to social (non-sexual) songs during the non-reproductive season, and higher in response to sexual

songs during the reproductive season. “
“Past studies in songbirds have highlighted a central role for the medial preoptic nucleus (mPOA) in context-appropriate vocal communication. During the breeding season, male songbirds sing primarily to attract females (sexually motivated song) and to repel competitors (agonistically motivated song). Past data have linked dopamine and D1 dopamine receptors in the mPOA to sexually motivated but not agonistically motivated song; however, direct effects of dopamine receptor manipulations in the mPOA on song have not been experimentally tested. Here, we tested the hypothesis that D1 receptor stimulation in the mPOA selectively influences sexually motivated male song, and the possibility that the effects of D1 receptor agonism differ at low and high doses. In a first study, breeding-condition male European starlings

received infusions of saline or a single dose of the D1 receptor agonist SKF 38393 on separate test days into the mPOA or hypothalamic for control areas. Stimulation of D1 receptors in the mPOA triggered sexually motivated but not agonistically motivated song. A second study showed inverted-U shaped dose–response effects of the agonist, such that low levels of sexually motivated song were observed at low and high levels of D1 receptor activation. A third study showed that the effects of the D1 receptor agonist were blocked by the D1 receptor antagonist SCH 23390. These findings suggest that an optimal level of D1 receptor stimulation in the mPOA is needed to facilitate sexually motivated vocal production.

Most studies conducted in HIV-infected individuals have evaluated immunological responses to one or two specific vaccines. There is very little information on humoral responses to a multiple vaccination programme and the maintenance of long-term antibodies in HIV-infected subjects. Moreover, there are few reports on the influence of highly active

antiretroviral therapy (HAART) and its interruption on specific vaccine immunological responses [9]. We carried out a double-blind, placebo-controlled clinical trial in 26 successfully treated HIV-1-infected adults attending selleck screening library the Hospital Clínic of Barcelona (Spain) between June 2003 and July 2006 in order to assess the safety and immunological effects of a multiple vaccination programme and

the influence of HAART and its interruption on vaccine-induced immunity. We designed a single-centre, prospective, randomized, double-blind placebo-controlled trial to assess the influence of a vaccination programme on viral load (VL) rebound and HIV-1-specific immune responses in successfully treated HIV-infected subjects [10]. Patients attending the Infectious Diseases Unit of the hospital were invited to participate in the study if they met the following inclusion criteria: asymptomatic this website HIV-1 infection, age ≥18 years, CD4 count ≥500 cells/μL, nadir CD4 count >300 cells/μL, plasma VL<200 HIV-1 RNA copies/mL and administration of HAART for at least 12 months. Exclusion criteria were baseline creatinine >2.5 mg/dL, Glutamic-Oxaloacetic Transaminase/Glutamic-Pyruvic Etomidate Transaminase (GOT/GPT)>250 IU/L, chronic hepatitis B, known allergy to a vaccine or vaccine component, pregnancy or planned pregnancy. Twenty-six subjects were enrolled and all received counselling on safe sexual practices. The study was approved by the Ethics Committee of the Hospital Clínic of Barcelona and was registered in the public clinical trials database of the National Institutes of Health (NIH; NCT00329251).

After giving written informed consent, patients were invited to visit the Adult Vaccination Center of the hospital where they were randomized to either the vaccine group (n=13) or the placebo group (n=13). The immunization programme included the following vaccines: hepatitis B (months 0, 1, 2 and 6), influenza (month 1), pneumococcal (month 2), hepatitis A (months 4 and 10), varicella (months 4 and 6), measles-mumps-rubella (month 8) and diphtheria-tetanus (month 10). The control group received injections containing placebo according to the same schedule. At month 12, HAART was discontinued for at least 6 months (month 18) and the evolution of the vaccine-induced immunity was analysed for the whole cohort and compared between groups.

Over 4 months in 2010, a health advisor (HA) approached 19–65-year-olds at a central London acute medical admissions unit and offered a rapid HIV point of care test (POCT) with the aid of an educational video. Feasibility and acceptability were assessed through surveys and uptake rates. Costs per case of HIV infection identified were established. Of the 606 eligible people admitted during the pilot, 324 (53.5%) could not be approached or were individuals for whom testing was deemed inappropriate. In total, 23.0% of eligible admissions had an HIV POCT. Of the patients who watched the GDC0199 video and had not recently been tested for HIV, 93.6% (131 of 140) agreed to an HIV test; four

further patients had an HIV test but did not watch the video. Three tests (2.2%; three AZD1208 cost of 135) were reactive and all were confirmed HIV positive on laboratory testing. HIV testing in this setting was felt to be appropriate by 97.5% of individuals. The cost per patient was £21, and the cost per case of HIV identified was £1083. Universal POCT HIV testing in an acute medical setting, facilitated by an educational video and dedicated staff, appears acceptable, feasible, effective, and low cost. These findings support the recommendation of HIV testing

for all medical admissions in high-prevalence settings, although with this model a significant proportion remained untested. The publication of the national guidelines on HIV testing [1] prompted a number of initiatives to assess the feasibility, acceptability and cost-effectiveness of new models of delivery for HIV testing. Our aim was to determine whether a model of care utilizing a multimedia tool and dedicated staff found to be effective in an emergency medical setting in New York [2, 3] is an acceptable, feasible L-gulonolactone oxidase and cost-effective model when translated to the UK. Data were collected on all admissions

to an acute admission unit (AAU) in Central London over a 16-week period in 2010. Adults aged 19–65 years in a stable condition were eligible for inclusion in the HIV testing pilot. Patients who were only admitted during the weekend were excluded from this analysis. The service model consisted of a health advisor (HA) approaching all stable admissions, and offering HIV testing with the aid of an educational video. If the patient accepted the offer of testing, a finger prick rapid HIV point of care test (POCT) was performed using the INSTI™ (bioLytical Laboratories Inc., Richmond, B.C., Canada) test [4]. If the result was HIV negative, the patient had the option of watching a post-test video providing risk-reduction information. If the result was reactive, the HA arranged confirmatory testing and urgent follow-up with the HIV service. All patients completed a questionnaire to evaluate patient satisfaction and collect process evaluation data. The questionnaire was delivered electronically via the laptop that patients used to watch the videos.

The analysis of the mutants should continue, especially with respect to changes in membrane properties caused by the presence and absence of the distinct modifications. Hand in hand should go a structural determination of the products of the OlsD- and OlsE-catalyzed reactions.

The exact structure of both modifications is required to understand the function/properties of the different lipids on a biophysical level. M.Á.V.-G. is a PhD student from the Programa de Doctorado en Ciencias Biomédicas, Universidad Nacional Autónoma de México, and is a recipient of a scholarship from the Consejo Nacional de Ciencia y Tecnología, México. Ivacaftor cost Work in our laboratory has been financed by grants from CONACyT-Mexico (46020-N and 153200) and DGAPA/UNAM (IN217907

and IN201310) to C.S. “
“Most of our limited knowledge of microbes in corals comes from stony and soft corals; the microbial diversity of black corals is still poorly understood. Microbial diversity of the South China Sea black coral Antipathes dichotoma was investigated using a culture-dependent method followed by analysis of bacterial 16S rRNA gene and fungal internal transcribed spacer sequences. A total of 36 bacterial and 24 fungal isolates were recovered and identified, belonging to three bacterial phyla (Firmicutes, Actinobacteria and Alphaproteobacteria) and four fungal orders (Eurotiales, Hypocreales, Pleosporales and Botryosphaeriales). The high level microbial diversity of A. dichotoma is in accordance with previous studies on those of some stony and soft corals. However,

the lack of bacterial Gammaproteobacteria phylum in A. dichotoma is in sharp contrast E7080 chemical structure to the stony and soft corals, in which the Gammaproteobacteria phylum is relatively common and abundant. Antimicrobial activities of 21 bacterial and 10 fungal representative isolates (belonging to 21 different bacterial and 10 different fungal species, respectively) were tested against two marine pathogenic bacteria and two marine coral pathogenic fungi. A relatively high proportion (51.6%) of microbial isolates displayed distinct antibacterial and antifungal activities, suggesting that the black mafosfamide coral-associated microorganisms may aid their host in protection against marine pathogens. This is the first report on the diversity of culturable microorganisms associated with black coral. It contributes to our knowledge of black coral-associated microorganisms and further increases the pool of microorganisms available for natural bioactive product screening. Coral reefs around the world are in decline, and infectious diseases are one of the main visible causes (Richardson & Aronson, 2002). As a result, more attention has been focused on the coral-associated microbes that may play a role in establishing diseases and the connections existing between the microbial communities and the overall health of the corals (Kellogg, 2004).

5 mM for NADPH and 0 to 5 mM for thio-NAD+. Least-squares fits to double reciprocal plots (Lineweaver–Burk plots) were used to calculate the apparent kinetic parameters. The effects of metal ions (NaCl, RbCl, KCl, LiCl, MgCl2, CaCl2, MnCl2, CoCl2, ZnCl2, NiCl2, CuCl2) on EcSTH activity were measured using two methods: first,

enzyme activity was determined in the standard reaction mixture supplemented with 2 mM metal ions; second, the enzyme was preincubated with 2 mM metal ions for 30 min at 4 °C and the activity was then assayed in a standard reaction mixture. The effects of adenine nucleotides (2 mM ATP, 2 mM ADP, 2 mM AMP), reducers [2 mM dithiothreitol (DTT), 0.2%β-mercaptoethanol], a chelating agent (2 mM MK-1775 solubility dmso EDTA) and a nonaqueous solvent [0.2% dimethyl sulfoxide (DMSO)] on EcSTH activity were Fulvestrant cell line tested using the same methods. A search of the KEGG Enzyme Database for enzymes with STH activity, and of GenBank using NCBI blast for sequences >40% similar

to E. coli sth, reveals that the enzyme is found far beyond the Gammaproteobacteria and a few mycobacteria as first reported (Boonstra et al., 2000b). Many actinobacteria and some members of the Alpha-, Beta-, Deltaproteobacteria and Spirochaetales all contain the enzyme. Moreover, microorganisms harboring two transhydrogenases are not only found in the enterobacteria (Boonstra et al., 1999; Sauer et al., 2004) but also in most organisms that contain the sth gene. Interestingly, plants seem not to have either transhydrogenase; perhaps, other unknown genes perform functions similar to sth or pntAB (Thompson et al., 1998; Bykova et al., 1999) ROS1 or perhaps unidentified mechanisms regulate

the balance between NAD(H) and NADP(H) pools (Sauer et al., 1997; Wittmann & Heinzle, 2002; Marx et al., 2003). A 1401-bp PCR product was amplified from E. coli MG1655 and cloned into pBluescript SK(+). Escherichia coli DH5α harboring pSTH was induced by IPTG to overexpress the fused EcSTH. The purified enzyme was homogeneous as judged by SDS-gel electrophoresis (Fig. 1a), and the molecular mass of each subunit, approximately 52 kDa, is consonant with the predicted molecular weight of EcSTH (51.5 kDa) and previous reports for STHs from Azotobacter vinelandii, E. coli and Pseudomonas fluorescens (French et al., 1997; Boonstra et al., 1999, 2000b). Western blot analysis reveals a single protein band using the anti-6 × His antibody as a probe (Fig. 1b). The effects of pH and temperature on EcSTH were determined in Tris-HCl buffer. The optimal pH of EcSTH is pH 7.5 (Fig. 2a), which is similar to the optimal pH for EcSTH cofactor regeneration (between pH 7.5 and 8.0; Ichinose et al., 2005; Mouri et al., 2009). The optimal temperature for catalysis by EcSTH is 35 °C (Fig. 2b). This result is similar to A. vinelandii STH (30–35 °C) (Chung, 1970). EcSTH is stable below 50 °C.