robusta homologues (i.e. ORF249) appear to be poorly

conserved, and are likely not functional. Fragments of the plasmid ORFs are also found in the gene-poor regions. Eight incomplete ORFs with similarity to C. fusiformis ORF482/ORF484, sometimes without start codon, are interspersed throughout region III and IV. One of the contigs with high read depth could not be assembled into the chloroplast genome. Upon closer analysis, this contig was found to constitute a separate circular molecule with a size of 3813 bp with significant similarity to C. fusiformis pCf2, which we designated as GPCR Compound Library solubility dmso pSr1 ( Fig. 3C). A previous survey did not identify any plasmids in two other members of the Naviculaceae, Fistulifera pelliculosa and Navicula incerta ( Hildebrand et al., 1991), and no plasmid was reported in Fistulifera sp. JPCC DA0580 ( Tanaka et al., 2011). Thus, the pSr1 plasmid is the first to be identified in a diatom belonging to Naviculales. Plasmids may not be a common feature in diatoms belonging to this order. Alternatively, plasmids have not been detected in previous studies due to technical limitations. Purification of chloroplast DNA by cesium chloride or sucrose gradient centrifugation may result in the loss of any associated plasmid DNA. pSr1 contains three ORFs encoding putative proteins of 494, 317

and 121 AAs, which show significant similarity to pCf2 ORF484, ORF246 and ORF125, respectively (NCBI BlastP expect value < 1e-36). The C-terminal part of pSr1 ORF317 also shows similarity to a small selleck chemicals GABA Receptor ORF in pCf2 (ORF64) that overlaps with pCf2 ORF246 (Fig. 3B). Introducing a deletion at position 732 of pCf2 ORF246 and an insertion in position 191

of ORF64 results in a continuous ORF encoding a putative protein of 311 AAs (ORF311) showing high similarity to pSr1 ORF317 and S. robusta chloroplast ORF292 ( Fig. 3B; Fig. A.3). The two frameshifts in pCf2 may be the result of sequencing errors. Alternatively, they have occurred as part of an inactivation of the ORF311 locus. The only C. fusiformis plasmid ORFs with a putative function are ORF217/ORF218, which show similarity to serine recombinases. Homologues of these ORFs are not found in pSr1; however, gene-poor region III in the chloroplast genome encodes a serine recombinase, termed SerC2, with similarity to CfORF217 and CfORF218 as well as K. foliaceum SerC1 and SerC2 and Fistulifera sp. SerC2. Residues found to be critical for the active site of serine recombinases (Arg-8, Ser-10, Asp-67, Arg-68 and Arg-71 in the Escherichia coli γδ resolvase ( Grindley et al., 2006)) are conserved in all diatom chloroplast serine recombinases. They also show a similar size and domain structure as γδ resolvase, suggesting that they may act through a similar mechanism. Although the intracellular localisation of pSr1 is not known, it appears to be closely associated with the chloroplast genome. Cloned pCf2 hybridised to both chloroplast and nuclear DNA from C. fusiformis ( Jacobs et al., 1992).

After 10 min, we examined the contralateral hemisphere with the same protocol. We selected ROIs in the contralateral MCA territory, which corresponded in size, shape and localization to the Galunisertib mw ischemic ROIs (Fig. 3). Parameters of refill kinetics (A, β and the product A × β) were extracted from each ROI for statistical analysis. To analyze the potential relationship between MCA flow velocity and the parameters

of refill kinetics, we subdivided patients in two groups: patients with persisting MCA pathology defined by COGIF grades of 3 or lower, and patients with symmetrical or increased MCA flow (COGIF grade 4). We examined 31 patients (17 male, 14 female, mean age 68.3 ± 13.4) who were admitted to our stroke unit with acute ischemic stroke in the MCA territory (Table 1). 58% of patients were treated with intravenous thrombolysis. At the time point of examination, TCCD showed a persistent pathological flow pattern of

the ipsilateral MCA (COGIF grades 0–3) in 21/31 (67.7%) patients. Pathological flow patterns were more frequent among patients who were not VX-809 mw treated with tPA (11/13 vs. 10/18, p = 0.08). Rt-UPI showed significantly lower values of the refill parameter β in the ischemic area compared to the contralateral MCA territory (β (1/s): 0.75 ± 0.41 vs. 1.05 ± 0.51, p < 0.05). The difference between ischemic and contralateral ROIs was more prominent in patients with persisting MCA obstruction (n = 21; β (1/s): 0.61 ± 0.31 vs. 1.01 ± 0.53, p = 0.005). Correspondingly, in patients with symmetrical or increased ipsilateral MCA SB-3CT flow, β values were not significantly different between both hemispheres (n = 10; β (1/s): 1.04 ± 0.47 vs. 1.14 ± 0.49, p = n.s.). There was no significant difference between β values of the ischemic tissue of patients treated with tPA and those who did not receive systemic thrombolysis (β (1/s): 0.72 ± 0.32 vs. 0.78 ± 0.53, p = n.s.). For the plateau of acoustic intensity (A) and the product of A and β

(A × β), there was a high interindividual variance of the values, resulting in no significant difference between ischemic or contralateral healthy tissue in any group of patients ( Table 2). This study investigated the feasibility of rt-UPI with refill kinetics to assess perfusion deficits related to persistent or already recanalized arterial obstruction in acute MCA stroke patients. The parameter β, which represents the slope factor of the exponential function of refill kinetics, shows overall significant differences between ischemic and healthy tissue. This finding was more pronounced in patients with COGIF grades 0–3 and was absent in COGIF grade 4. The parameters A and the product A × β showed high standard deviations in our study, which resulted in a lack of significance between ischemic and non-ischemic tissue for these parameters.

p). Hesperidin powder was dissolved in 0.1% carboxy methyl cellulose and each rat received

daily 1 ml at a dose of 20, 40 and 80 mg/kg body weight orally by intragastric tube throughout the experimental period. The animals were randomly divided into six groups of six rats in each group. Group I: served as control (isotonic saline). Group II: animals were orally administered with hesperidin alone (80 mg/kg body weight). Group III: animals received ferrous sulfate (30 mg/kg body weight). Group IV-VI: animals were treated with ferrous sulfate (30 mg/kg body weight) following oral administration of hesperidin (20, 40, 80 mg/kg body weight) for 10 days. At the end of the experimental period, animals in different groups were sacrificed by cervical decapitation. Blood samples were collected without heparin for serum separation. Serum separated by centrifugation was used for various biochemical estimations. Rats were anesthetized by ketamine (28 mg/kg buy A-1210477 body weight, intra muscularly) and the animals were sacrificed by cervical decapitation. The liver and kidney was quickly excised, rinsed with isotonic saline, blotted dry on filter paper, weighed and then 10% (w/v) homogenates of tissue was prepared in buffer (0.1 M Tris-HCL buffer (pH 7.4) and centrifuged at 3000 × g for 20 min at 4 °C. The resulting tissue homogenate was used for various biochemical assays. The activities of serum aspartate aminotransferase

(E.C.2.6.1.1), alanine aminotransferase (E.C.2.6.1.2), alkaline phosphatase (E.C.3.1.3.1) and lactate dehydrogenase (E.C.3.1.3.1) were assayed using commercially LY2109761 in vivo available diagnostic kits (Sigma diagnostics (I) Pvt. Ltd., Baroda, India). Gamma glutamyl transferase (E.C.2.3.2.2) activity was determined by the method of Rosalki et al., 1970 [16] using γ-glutamyl-p- nitroanilide as substrate. Based on Vanden Berg reaction, serum second bilirubin was estimated by the method of Malloy and Evelyn, 1937 [17]. The activities of urea, creatinine and were estimated by Agappe Diagnostic (I) Pvt. Ltd., Kerala, India. Haemoglobin was estimated

by Drabkin and Austin, 1932 [18]. Creatinine clearance as an index of glomerular filtration rate was calculated from creatinine level in serum and creatinine level in 24 h urine sample. For determination of iron in blood, 1 ml of blood was digested with nitric acid in microwave oven. After digestion, iron was continuously pre concentrated and determined by flame atomic absorption spectrophotometry. A Perkin-Elmer 5000 atomic absorption spectrometer furnished with an iron hollow-cathode lamp (lamp current 4 mA) was used to determine the iron concentration. The instrument was set at 228.8 nm with a slit width of 0.5 nm. The acetylene flow rate was 2.0 l/min and an airflow rate of 17.0 l/min was employed to ensure an oxidizing flame. Lipids extracted from the tissues using by the method of Folch et al., 1957 [19].

The hydrological droughts on daily time scale at low truncation levels

such as Q90, Q95 have Dolutegravir in vitro also been attempted on non-stationary daily flows using the frequency analysis of observed durations and magnitudes (Zelenhasic and Salvai, 1987 and Tallaksen et al., 1997). Although the assessment and prediction of meteorological droughts on weekly time scale have been practiced using the Palmer Drought Severity Index (PDSI) or Standardized Precipitation Index (SPI), in literature only a few studies on the modeling of hydrological droughts on weekly time scale have been reported. The analysis of hydrological droughts on weekly time scale is desirable because effects of droughts are more palpable in agricultural production, municipal water supplies, small-scale hydro generation etc. The development of suitable predictive and assessment tools for hydrologic droughts at weekly time scale would be useful in managing available water resources

and off-setting effects of droughts. This paper attempts to develop suitable methodology to analyze and predict hydrological droughts at weekly time scale. The paper also embodies the results of drought models for comparative purposes at annual and monthly time scales in Canadian learn more streamflows. It has been observed (Bonacci, 1993, Woo and Tarhule, 1994, Sharma, 1997 and Sharma, 2000) that in general the drought intensity (I, i.e. MT = I × LT) is poorly Resveratrol correlated to LT. In view of a poor correlation (i.e. near independence) between these

two entities, the above relationship can be expressed in terms of expectations as E(MT) = E(I) × E(LT), which allows the prediction of drought magnitude with a priori knowledge of drought length. The drought intensity (I) can be modeled satisfactorily by the truncated normal distribution of SHI values which are laying below the truncation level. The modeling of drought length or duration (LT) is therefore essential in addressing the issues related to hydrological droughts. In the past, the theorem of extremes of random numbers of random variables ( Todorovic and Woolhiser, 1975; referred to hereafter as the extreme number theorem) has been used to model LT on annual flow series ( Sen, 1980a, Sharma, 1997, Sharma, 1998, Sharma, 2000, Panu and Sharma, 2002 and Panu and Sharma, 2009) and monthly flow series ( Sharma and Panu, 2008). Further, Sharma and Panu (2010) noted that the above theorem breaks down when the SHI sequences are strongly dependent (i.e. lag-1 autocorrelation being above 0.50) to the first order and/or extend to the second or higher order dependence (in case of weekly time scale). The monthly and weekly SHI sequences exhibit this tendency when the rivers are originating in lakes or passing through them. Under these circumstances, a second order Markov chain model tends to recover the analysis for modeling LT.

Once death was confirmed the pulmonary system was flushed with a heparin-solution (Wockhardt UK Ltd., Wrexham, UK) via catheter inserted into

the right ventricle or caudal vena cava. This was followed by Dublecco’s phosphate buffer solution (D-PBS, Sigma–Aldrich ABT-737 research buy Ltd., Gillingham, UK) to remove remaining blood from circulation. The lungs were inflated with around 3 ml of air and the trachea clamped; then the lungs, heart, and connective tissue were extracted en bloc. After extraction the lung’s trachea was cannulated and a syringe was used to breathe the lungs to ensure that they did not leak. Lungs were stored in glucose solution (5% glucose in water, Baxter Healthcare Ltd., Thetford, UK), chilled XL184 to approximately 280 K until needed. Excised rat lungs were inserted into a custom-made, sealable, ventilation chamber that filled the entire coil region. The ventilation chamber and its operating procedures are described in detail in previous work [15]. Briefly, the trachea of the rat lung was cannulated with an adaptor that was attached to the top of the ventilation chamber. The ventilation chamber was filled to about 2/3 of its total volume with a 5% glucose solution (Baxter Healthcare Ltd., Thetford, UK). Hp gas was delivered to the storage volume VB after compression using one of the two Extraction

Schemes described in this work. When a volume was pulled on the inhalation syringe pressure equalization forces the lungs to expand ( Fig. 8). This acts in a similar fashion to the thoracic diaphragm, as the expansion of the lungs causes it to inhale Fossariinae gas from the volume VB. Rubidium filters were made from 60 mm

of Teflon tubing (outer-diameter = 9.4 mm, inner-diameter = 6.4 mm; Swagelok, Warrington, UK) with 100 g of glass wool (Corning glass works, Corning, NY, USA) loosely packed inside. Chemical indicator paper (Whatman plc, Maidstone, UK) was used to check the pH value of the 1.0 ml of distilled water used to wash the glass wool. The resulting pH of the rubidium wash was pH 5.0. After SEOP at 220 kPa, a transfer of 5 s in duration resulted in a pressure of approximately 11 kPa of hp gas in Vext. Valves A + B ( Fig. 3a) were closed and the connecting lines were evacuated. A selected pressure of O2 gas was then added to Vext and the connecting lines were evacuated again. After a 5 s time delay that allowed for mixing of the O2 with the hp gas, the mixture was delivered for the MR measurements performed using Extraction Scheme 2. All T  1 data were obtained at ambient temperature using a pulse sequence comprising of sixteen medium (θ=12°)(θ=12°) flip angle r.f. pulses evenly separated by time increment τ. T1 relaxation values were determined from the nonlinear least-square analysis of the time dependence of the NMR signal intensity f(t) in the presence of spin-destruction due to the r.f.

Respiratory distress syndrome was the most frequent in children with DD (n=8; 67%) and CP (n=12; 60%). Also congenital pneumonia was most often diagnosed in children with DD (n=8; 67%) and CP (n=10; 50%), whereas less frequently – in

children with PE. BPD was the most frequent in the group of children with CP (n=7; 35%). A congenital heart defects were most frequent in the group of children with CAODS (82%), whereas they did not occur in the groups with PE, DD and ND. A relatively high incidence of respiration distress in the neonatal period in children with CAODS may be associated with the presence of other congenital defects (mainly heart, respiratory and gastrointestinal system). Perinatal pathology was not observed in the group with neuromuscular diseases (Tab. II). Among factors determining the recurrence of respiratory Bosutinib order tract infections resulting from a neurological condition: muscular hypotonia, weakness of respiratory muscles, adverse drug reactions (some antiepileptic and myorelaxant medicines) were analysed. Muscular hypotonia occurred most often in children with ND (n=5; 95%) and with EP (n=18; 85%); least frequent was in patients with CP. Chest deformation was most often observed in the group with ND (n=4; 66%), least frequent was in the group with DD (n=2; 16%); in the other groups chest deformities

were found in approx. 40% of children. The antiepileptic and CYTH4 myorelaxant drugs (mainly benzodiazepines and phenobarbital) were applied JNK high throughput screening in children in all groups, except for ND, most often in patients with CP and PE (Tab. III). The factors promoting recurrent infections of the lower respiratory tract include:

the body mass deficiency (most severe in the groups with PE; n=17; 74% and CAODS; n=8; 73%), gastroesophageal reflux and hypoproteinemia. GER was most frequently diagnosed in children with DD (n=8; 67%) and with PE (n=11; 48%). A high GER incidence in the first group may be connected with the age range and the existence of physiological reflux and in the patients. Hypoproteinemia was most often observed in the group with PE (n=10; 43%). An important factor responsible for the recurrence of respiratory tract infections is colonization of the airways by pathogenic flora. Such colonization was most often observed in children with neuromuscular diseases, which mainly resulted from a long-term course of the underlying disease and frequent hospitalizations (also in the intensive care units), due to a severe course of infections (Tab. IV). The relapses of lower respiratory tract infections in children with neurological diseases manifested as respiration disorders with dyspnoea. Radiologically confirmed pneumonia was most often diagnosed in children with PE and ND. Dyspnoea was the least frequent in children with CP.

Ethanolic formulations of propolis

prevent its consumption by people who can not consume alcohol for medical reasons, such as diabetic patients. Several patents have dealt therefore with new methods or solvents besides ethanol to extract propolis (Kasuma and Kenichi, 2001a, Kasuma and Kenichi, 2001b and Namiki et al., 2005). These patents have reported the use of edible vegetable oils, triglycerides selleck inhibitor and fatty acids as extraction solvents for propolis. Data on the biological activity and chemical composition of oil extracts of propolis are, however, scarce. Tosi, Donini, Romagnoli, and Bruni (1996) evaluated the antimicrobial activity of commercial extracts of propolis prepared with different solvents including oils. They reported a wide range of antimicrobial activity for the oil extract and concluded that the solvent employed for the extraction of propolis influences the potency of its antimicrobial activity.

We have compared antiproliferative activity against the HL-60, MDAMB-435 and SF-295 cells lines of oil and ethanolic propolis extracts (Buriol et al., 2009) and found out that oil extracts were active against the tumour cell line tested showing higher anticancer potential against the SF-295 cell line. The aim of this study was to investigate the effects of the oil and ethanolic extracts of propolis in experimental models. Hematological, biochemical, histopathological and morphological analyses of the tumour MTMR9 and the organs, including liver, spleen AZD2014 supplier and kidney, were performed to evaluate the toxicological aspects of the treatment. The results of this study should

therefore advance the knowledge of the antitumour benefits of edible oil extracts of propolis and provide a better understanding of their application in the prevention/treatment of malignant tumours. Besides biological assays, the oil extract of propolis was also fractioned by chromatography and its fractions analysed using mass spectrometry to evaluate their chemical composition. Propolis samples were collected in 2006 and supplied by Campolin and Schmidt Company from Prudentópolis city (Paraná State, Brazil). Propolis was stored at −18 °C until extraction. Fifty grams of propolis were extracted in a shaker with 500 ml of canola oil or 70% ethanol, during 24 h, at room temperature. After that period, the extractive solutions were filtered. The solvent was removed from the hydro-alcoholic solution yielding EEP70. The oil extract of propolis was partitioned into 80% v/v methanol/water and the aqueous methanolic phase was dried in a rotatory evaporator yielding ODEP. Hundred and eighty milligrams of ODEP in methanol were applied on a glass column (3 × 80 cm) containing Sephadex LH-20. The elution was performed with methanol. A total of 80 fractions of 8 ml each were obtained.

For validation purposes, five liver extracts with low recoveries were diluted up to 1000 times and analyzed on a Xevo TQ-S mass spectrometer (Waters Corporation, Milford, USA), which is a more sensitive instrument compared to the Quattro Premier find more XE. The recoveries of 13C4-PFOS increased from 10–44% to 36–80% in the × 100 and × 1000 diluted samples (Fig. S1, Supplementary data). To compare PFOS concentrations in undiluted (u) and diluted (d) extracts, the mean normalized difference (%) was calculated using the formula: ((u − d) / ((u + d) / 2) × 100). The calculated concentrations

of all the diluted extracts, except for one sample, were well in range with the initial concentrations (average mean normalized difference of 18%). Consequently, reliable results can be produced even when recovery rate is low since the internal standard and the native compound are equally suppressed. Recoveries, method reproducibility and method detection limits (MDL) for all samples are presented

in Table S1, Supplementary data. One milliliter of ultra pure water was used as procedural blanks and extracted in the same way as the real samples. The MDL was defined as the mean concentration in the procedural blanks plus three standard deviations, and the limit of detection (LOD) for individual samples was calculated as three times the noise level. Overall good recoveries (> 50%) of 13C-PFOS and 13C-PFOA were measured for the samples after the replacement of the recovery standard 7H-PFHPA

to 13C8-PFOS and 13C8-PFOA in the middle of the project. One two year old mink TSA HDAC mouse caught in autumn in the G area was excluded due to non-reproducible results of the diluted extracts. Using the general linear model (GLM) procedure of SAS (SAS Institute Inc., Cary, NC, USA, version 9.02.01), a multiple regression model with the concentrations Temsirolimus mouse of PFHxS, PFOS, PFNA, PFDA or PFUnDA as dependent variable and the sample area, sample season, age, body condition, year (of capture) and body weight as independent variables were elaborated on. PFBS, PFOA, PFDoDA and PFTrDA were excluded from this model since the concentrations were consistently low (< 17 ng/g). The model was fitted manually, starting with all variables in the model. Variables that were unsignificant (p > 0.05) for all dependent variables were removed. Relevant interactions between the effects were tested but none were included in the model due to insignificance or small sample size. The variable age was tested in several ways (different assignments into categories and numerical approaches), but had no significant effect. Area and season were the only variables that had a significant effect and were therefore the only variables kept in the final model: Y=μ+AREA+SEASON+ERROR.Y=μ+AREA+SEASON+ERROR.

Sweden is an ideal case for such an analysis since the retention approach has been practiced in this country for more than two decades (Eckerberg, 1988, Götmark et al., 2009 and Gustafsson and Perhans, 2010), and an extensive and high-quality National Forest Inventory data-base exists that can be used for detailed analysis. Due to a long history of industrial forestry in North Europe, and especially in Sweden and Finland, production forests have become more even-aged and much less structurally diverse than intact forests. Amounts of dead wood, old trees and other properties of importance to biodiversity are much lower

compared with natural forest landscapes (Fridman Natural Product Library and Walheim, 2000, Peterken, 2001 and Josefsson and Östlund, 2011). The importance of incorporating old-growth elements in managed forests is increasingly being recognized

(e.g. Bauhus et al., 2009), and dead trees and old living trees are known to be of large importance to biodiversity, not the least to threatened species (e.g. Bernes, BIBF1120 2011). A multiscale model for forest conservation is applied in Sweden, implying that conservation actions are taken at different scale-levels from individual trees to areas embracing hundreds or thousands of ha. The highest level, up to 1000 ha or more, includes formally protected areas such as national parks and nature reserves. At the next, intermediate level (ca. 1–50 ha) there are both formally protected and voluntary set-asides through certification, many many of which are so called woodland key habitats (Timonen et al., 2010). Retention approaches represent the lowest scale level, implying that trees of importance to biodiversity and ecosystem function

are left unlogged, mainly at final felling operations, but also during thinning. Single living trees are retained, and tree patches may be left as ‘islands’ in felled areas or adjacent to non-felled stands, often as buffer strips along lakes, rivers, wetlands and near settlements. Standing and lying dead trees are also retained, and according to instructions they should not be harmed during logging operations. Dead wood is created, in Sweden primarily through artificially creating snags by cutting trees at a height of 3–4 m, but also by retaining living trees of which some or many will eventually become windthrown. The state is responsible for establishing nature reserves, while both the state and the forest owners protect also smaller areas. Retention requirements have been part of Swedish forest legislation since the 1970s, and were made well-known to landowners through the “Richer Forest” campaign by the Swedish Forest Agency in the beginning of the 1990s. They were further consolidated with the launch of a new forest policy in the mid 1990s in which environmental and production goals were assigned equal value (Bush, 2010).

, 1996, Namkoong, 2002, McKinnel, 2002, Bariteau, 2003 and Aravanopoulos, 2011) and much scientific attention has been paid to evolutionary and adaptive processes (e.g. Eriksson et al., 1993; Namkoong et al., 2002; Le Corre and Kremer, 2003 and Le Corre and Kremer, 2012) as a basis. However, a general application and scaling-up of the verifiers

proposed by Namkoong TSA HDAC molecular weight et al. (2002) have not yet been feasible due to the difficulties summarized above. Any relevant set of indicators for trends in genetic diversity must include components at different scales (local/landscape/national/regional/global), involving the amount of diversity and how it is distributed in space. There is a need to identify genetically appropriate indicators and, at the same time, not to inflate the already large number of indicators that exist at global and regional scales. The State–Pressure–Benefit–Response (S–P–B–R) loop developed by UNEP/CBD/AHTEG, 2011a and UNEP/CBD/AHTEG,

2011b and Sparks et al. (2011) provides a well-considered and appropriate framework to ensure that the suggested set of indicators meet the requirements of being scientifically sound, selleckchem realistic, and policy relevant; and the framework has been adopted for implementation by BIP, 2013. The identification of indicators of tree genetic diversity should therefore preferably take place within such a framework and result in a set of S–P–B–R indicators. In Table 5 we list what we consider to be relevant operational indicators

and their type (state, pressure, benefit, response) at different geographic levels (global, regional/national and local) under the headline indicator trends in genetic diversity of tree species. Our table is not necessarily exhaustive, but proposes a fairly complete set of indicators all and has been made in congruence with Table 2. However, no separate pressure indicators are identified. Pressure indicators of genetic diversity are intrinsically linked with state indicators and the identification of the impact of any kind of pressure will have to rely on the knowledge of the state. Response indicators are referred to as response–benefit, because the rationale for a response is typically based on benefit. In Table 5 we subdivide the headline indicator trends in genetic diversity of tree species into seven operational indicators. These are appraised based on 21 verifiable indicators using a total of 34 verifiers. Genetic diversity indicators that are proposed in order to assess the adaptive potential of forest tree species from the global to the local level present different characteristics, such as indicator classification (state, pressure, benefit, response), reference level (global, regional, national, local), type of work needed (field, lab, web-based search, etc.), feasibility and type of expertise (direct measurement, or based on experimental analysis), level of informativeness, and cost.