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Abstract The expression of genes encompasses their transcription into mRNA followed by translation into protein. In recent years, next-generation sequencing and mass spectrometry methods have profiled DNA, RNA and protein abundance in cells. However, there are currently no reference standards that are compatible across these genomic, transcriptomic and proteomic methods, and provide an integrated measure of gene expression. Here, we use synthetic biology principles to engineer a multi-omics control, termed pREF, that can act as a universal molecular standard for next-generation sequencing and mass spectrometry methods. The pREF sequence encodes 21 synthetic genes that can be in vitro transcribed into spike-in mRNA controls, and in vitro translated to generate matched protein controls. The synthetic genes provide qualitative controls that can measure sensitivity and quantitative accuracy of DNA, RNA and peptide detection. We demonstrate the use of pREF in metagenome DNA sequencing and RNA sequencing experiments and evaluate the quantification of proteins using mass spectrometry. Unlike previous spike-in controls, pREF can be independently propagated and the synthetic mRNA and protein controls can be sustainably prepared by recipient laboratories using common molecular biology techniques. Together, this provides a universal synthetic standard able to integrate genomic, transcriptomic and proteomic methods.

Adding library adaptors to DNA samples is an essential step in preparing samples for next-generation sequencing. Here, Gunter et al. describe the development of Control Library Adaptors (CAPTORs), that correct sequencing errors and normalise quantitative biases in Nanopore libraries.

The Mexican fruit fly, Anastrepha ludens, is an important pest that causes widespread damage to a number of fruit crops in Mexico. The sterile insect technique (SIT) is commonly used for its control. However, the existence of natural barriers can give rise to a population structure in neutral loci and possibly behavioral or adaptive traits that interfere with SIT. For this reason, it is important to understand the genetic diversity and structure of A. ludens populations and to better understand the evolutionary ecology and population processes in view of possible expansions and possible host shifts due to climate change. We genotyped nine nuclear DNA (nDNA) microsatellite loci among fruit fly populations collected from five biogeographic areas within Mexico, namely, the Mexican Plateau, the Northeastern Coastal Plain, the Pacific Coast, the Gulf Coast of Mexico, and the Soconusco, and a laboratory strain. The nuclear genetic diversity was moderate (from He = 0.34 to He = 0.39) within the wild mexfly population. We found that populations were clustered in three genetic groups (K = 3). The diversity and the genetic structure of A. ludens are determined by environmental and geological conditions, as well as local conditions like anthropogenic perturbation, which would produce population expansion and the existence of possible predators that would affect the population density. Gene flow showed recent migration among populations. The laboratory strain showed fewer diversity than the wild samples. Large values of current and ancestral population size suggest high resistance to climatic changes, probably due to biological attributes, such as its polyphagous, multivoltine, and high dispersal characteristics. In particular, ecosystem fragmentation and perturbation as well as the existence of new plant hosts would probably increase the abundance of flies.

A massive mangrove dieback event occurred in 2015–2016 along ∼1000 km of pristine coastline in the Gulf of Carpentaria, Australia. Here, we use sediment and wood chronologies to gain insights into geochemical and climatic changes related to this dieback. The unique combination of low rainfall and low sea level observed during the dieback event had been unprecedented in the preceding 3 decades. A combination of iron (Fe) chronologies in wood and sediment, wood density and estimates of mangrove water use efficiency all imply lower water availability within the dead mangrove forest. Wood and sediment chronologies suggest a rapid, large mobilization of sedimentary Fe, which is consistent with redox transitions promoted by changes in soil moisture content. Elemental analysis of wood cross sections revealed a 30- to 90-fold increase in Fe concentrations in dead mangroves just prior to their mortality. Mangrove wood uptake of Fe during the dieback is consistent with large apparent losses of Fe from sediments, which potentially caused an outwelling of Fe to the ocean. Although Fe toxicity may also have played a role in the dieback, this possibility requires further study. We suggest that differences in wood and sedimentary Fe between living and dead forest areas reflect sediment redox transitions that are, in turn, associated with regional variability in groundwater flows. Overall, our observations provide multiple lines of evidence that the forest dieback was driven by low water availability coinciding with a strong El Niño–Southern Oscillation (ENSO) event and was associated with climate change.

One of Earth's foremost ecological challenges is the degradation of land habitats. This degradation is often caused by deforestation and desertification resulting from the unsustainable management of natural resources. Land restoration seeks to reverse this trend and repair ecosystems to better health. Indigenous peoples and local communities have a key role in realizing long-term, sustainable land restoration. Local and indigenous communities often have intimate knowledge of the local ecosystems and an interest in preserving ecosystem services. Areas managed by indigenous peoples and local communities especially overlap with remaining intact ecosystems and suffer from less deforestation than unprotected areas. Here, we discuss how the knowledge and engagement of local communities can improve the management, implementation, and monitoring of habitat restoration. However, there are also challenges to land restoration, and scientists and policymakers that can align restoration outcomes with community benefits gained from environmental stewardship and knowledge, are more likely to achieve long-term sustainable restoration success.

BackgroundThe province of Formosa, Argentina, is endemic for leprosy. In the present paper, we assessed the trend (T, 2002-2016 time series) and the forecast for 2022 of new case detection rate (NCDR) and determined the spatial distribution of new cases detected (NCD) of leprosy.Methodology/principal findingsThis is a descriptive observational study of 713 NCD of leprosy from provincial medical records between January 2002 and December 2016. The whole dataset from the provincial medical record was used to independently estimate the NCDR trends of the general population, age groups, sexes and Departments. This same database was used to estimate the NCDR forecast of the general population for 2022, applying a dynamic linear model with a local linear trend, using the MCMC algorithm. The NCDR was higher in men (p65:-61, p0.05), which showed no trend. Forecasts predicted that leprosy will not be eliminated by 2022 (3.64, 95%CI: [1.22, 10.25]).Conclusions/significanceOur results highlight the status of leprosy in Formosa and provide information to the provincial public health authorities on high-risk populations, stressing the importance of timely detection of new cases for further elimination of the disease in the province.

Montane ecosystems occur throughout the world, and harbor many endemic species. They also provide key ecological services, including the catchment of water resources and the storage of organic carbon. These ecosystems are vulnerable to global climate change and increasing human pressures, including forestry and their conversion to arable land. In the extensive and biodiverse Mexican montane regions, ongoing deforestation and conversion to arable lands has led to diminished ecosystem health and services. Here, we undertook a comprehensive evaluation of carbon stocks within Mexican montane habitats in the Flora and Fauna Conservation Area of Nevado de Toluca. This aimed to integrate these habitats into Mexican and global census of forest carbon, the first step needed to convert on carbon credit markets to incentivize conservation of this region by local communities. Our study evaluated both, living biomass and belowground soil organic carbon in sites within forests, alpine grasslands and converted arable land. We addressed the following questions: (1) What are the organic carbon stocks, including the soil component, of our studied montane habitats? (2) What are the avoided CO2 emissions from maintaining natural forests and preventing conversion to arable land? And (3) Within our study area, are organic carbon stocks in the soil correlated to carbon stocks in aboveground living biomass? We found whole ecosystem organic carbon stocks ranged from 68 Mg OC ha–1 in unburnt alpine grasslands to 668 Mg OC ha–1 in Abies religiosa forests. By avoiding conversion of the A. religiosa forests to arable lands, we show that emissions of 1,122 to 1,671 Mg CO2 ha–1 are avoided. Notably, the belowground soil organic carbon stock comprised ≥ 40% of the total ecosystem organic carbon stock. We recommend soil organic carbon stocks should be included within Mexican and global forestry carbon stock inventories, and should be considered within voluntary carbon-credit markets used to incentivize the conservation of Mexican montane habitats.

Complex microbial communities pose a challenge to metagenomic analysis. Here the authors develop ‘sequins’, internal DNA standards that represent a synthetic community of artificial genomes.

Tree stems swell and shrink daily, which is thought to reflect changes in the volume of water within stem tissues. We observed these daily patterns using automatic dendrometer bands in a diverse group of mangrove species over five mangrove forests across Australia and New Caledonia. We found that mangrove stems swelled during the day and shrank at night. Maximum swelling was highly correlated with daily maxima in air temperature. Variation in soil salinity and levels of tidal inundation did not influence the timing of stem swelling over all species. Medium-term increases in stem circumference were highly sensitive to rainfall. We defoliated trees to assess the role of foliar transpiration in stem swelling and shrinking. Defoliated trees showed maintenance of the pattern of daytime swelling, indicating that processes other than canopy transpiration influence the temporary stem diameter increments, which could include thermal swelling of stems. More research is required to understand the processes contributing to stem shrinking and swelling. Automatic Dendrometer Bands could provide a useful tool for monitoring the response of mangroves to extreme climatic events as they provide high-frequency, long-term, and large-scale information on tree water status.

Background Dengue is a major and rapidly increasing public health problem. In Argentina, the southern extreme of its distribution in the Americas, epidemic transmission takes place during the warm season. Since its re-emergence in 1998 two major outbreaks have occurred, the biggest during 2016. To identify the environmental factors that trigger epidemic events, we analyzed the occurrence and magnitude of dengue outbreaks in time and space at different scales in association with climatic, geographic and demographic variables and number of cases in endemic neighboring countries. Methods Information on dengue cases was obtained from dengue notifications reported in the National Health Surveillance System. The resulting database was analyzed by Generalized Linear Mixed Models (GLMM) under three methodological approaches to: identify in which years the most important outbreaks occurred in association with environmental variables and propose a risk estimation for future epidemics (temporal approach); characterize which variables explain the occurrence of local outbreaks through time (spatio-temporal approach); and select the environmental drivers of the geographical distribution of dengue positive districts during 2016 (spatial approach). Results Within the temporal approach, the number of dengue cases country-wide between 2009 and 2016 was positively associated with the number of dengue cases in bordering endemic countries and negatively with the days necessary for transmission (DNT) during the previous autumn in the central region of the country. Annual epidemic intensity in the period between 1999–2016 was associated with DNT during previous autumn and winter. Regarding the spatio-temporal approach, dengue cases within a district were also associated with mild conditions in the previous autumn along with the number of dengue cases in neighboring countries. As for the spatial approach, the best model for the occurrence of two or more dengue cases per district included autumn minimum temperature and human population as fixed factors, and the province as a grouping variable. Explanatory power of all models was high, in the range 57–95%. Discussion Given the epidemic nature of dengue in Argentina, virus pressure from endemic neighboring countries along with climatic conditions are crucial to explain disease dynamics. In the three methodological approaches, temperature conditions during autumn were best associated with dengue patterns. We propose that mild autumns represent an advantage for mosquito vector populations and that, in temperate regions, this advantage manifests as a larger egg bank from which the adult population will re-emerge in spring. This may constitute a valuable anticipating tool for high transmission risk events.

Riverine wetlands are created and transformed by geomorphological processes that determine their vegetation composition, primary production and soil accretion, all of which are likely to influence C stocks. Here, we compared ecosystem C stocks (trees, soil and downed wood) and soil N stocks of different types of riverine wetlands (marsh, peat swamp forest and mangroves) whose distribution spans from an environment dominated by river forces to an estuarine environment dominated by coastal processes. We also estimated soil C sequestration rates of mangroves on the basis of soil C accumulation. We predicted that C stocks in mangroves and peat swamps would be larger than marshes, and that C, N stocks and C sequestration rates would be larger in the upper compared to the lower estuary. Mean C stocks in mangroves and peat swamps (784.5 ± 73.5 and 722.2 ± 63.6 MgC ha−1, respectively) were higher than those of marshes (336.5 ± 38.3 MgC ha−1). Soil C and N stocks of mangroves were highest in the upper estuary and decreased towards the lower estuary. C stock variability within mangroves was much lower in the upper estuary (range 744–912 MgC ha−1) compared to the intermediate and lower estuary (range 537–1115 MgC ha−1) probably as a result of a highly dynamic coastline. Soil C sequestration values were 1.3 ± 0.2 MgC ha−1 yr−1 and were similar across sites. Estimations of C stocks within large areas need to include spatial variability related to vegetation composition and geomorphological setting to accurately reflect variability within riverine wetlands.

Aim Implant rehabilitation has become a very reliable and safe procedure. However, in some cases, a small amount of bone could make implant surgery extremely difficult or even impossible. Hence, a surgical technique to augment sinus floor has been developed and improved. Nevertheless, there is still controversy over the use of a membrane over the osteotomy window. Therefore, the aim of this study was to investigate whether the use of a membrane could be beneficial in sinus floor augmentation. Materials and methods A group of 12 patients requiring sinus floor lift were recruited. The patients were randomly allocated to either control group (membrane) or test group (no membrane) and only one sinus for patient was augmented. After 6 months, a bone biopsy was harvested from the lateral window to be processed for histological analysis. Results The mean amount of newly formed bone in test group was 28.0±19.5%, the connective tissue accounted for a mean value of 59.2±15.6%, while 12.8±12.6% was the amount of residual graft particles. In the membrane group the newly formed bone counted for a mean value of 30.4±15.8%, the mean quantity of connective tissue was 50.3±18.9% and about residual graft particles a mean value of 18.2±20.4% was registered. Conclusion According to our data, the use of a membrane over the lateral bone wall in sinus lift surgery does not significantly influence healing. However, the membrane could influence the residual particles resorption rate as well as soft tissue ingrowth.

Numerous protocols have been published for extracting DNA from phlebotomines. Nevertheless, their small size is generally an issue in terms of yield, efficiency, and purity, for large-scale individual sand fly DNA extractions when using traditional methods. Even though this can be circumvented with commercial kits, these are generally cost-prohibitive for developing countries. We encountered these limitations when analyzing field-collected Lutzomyia spp. by polymerase chain reaction (PCR) and, for this reason, we evaluated various modifications on a previously published protocol, the most significant of which was a different lysis buffer that contained Ca2+ (buffer TESCa). This ion protects proteinase K against autolysis, increases its thermal stability, and could have a regulatory function for its substrate-binding site. Individual sand fly DNA extraction success was confirmed by amplification reactions using internal control primers that amplify a fragment of the cacophony gene. To the best of our knowledge, this is the first time a lysis buffer containing Ca2+ has been reported for the extraction of DNA from sand flies.

A fault with two asperities is modelled as a system made of two blocks coupled by a spring and sliding on a plane under the same values of static and dynamic friction. An analytical solution is given for the simultaneous motion of the blocks and the corresponding orbits are plotted in the phase space. It is proven that, whichever the initial state is, the long-term behaviour of the system is one of an infinite number of limit cycles, characterized by a particular pattern of forces. The region where the system is located when the blocks are stationary can be divided into narrow stripes corresponding to different orbits of the points belonging to them. This implies that the system is sensitive to perturbations and has relevant implications for a fault, which is subject to stress transfers from earthquakes generated by neighbouring faults. In this case, the fault may experience a larger earthquake, with the simultaneous failure of the two asperities, which restores a stress distribution compatible with periodic behaviour. The seismic moment associated with simultaneous asperity failure is always greater than the maximum value that can be released in a limit cycle. For strongly coupled asperities, the moment can be several times larger.

A system made of two sliding blocks coupled by a spring is employed to simulate the long-term behaviour of a fault with two asperities. An analytical solution is given for the motion of the system in the case of blocks having the same friction. An analysis of the phase space shows that orbits can reach a limit cycle only after entering a particular subset of the space. There is an infinite number of different limit cycles, characterized by the difference between the forces applied to the blocks or, as an alternative, by the recurrence pattern of block motions. These results suggest that the recurrence pattern of seismic events produced by the equivalent fault system is associated with a particular stress distribution which repeats periodically. Admissible stress distributions require a certain degree of inhomogeneity, which depends on the geometry of fault system. Aperiodicity may derive from stress transfers from neighboring faults.

La presente nota informa sobre 5 nuevos registros de especies de Phlebotominae para Argentina: Micropygomyia oswaldoi (Mangabeira) y Pintomyia bianchigalatiae (Andrede-Filho, Aguiar, Dias & Falcão) en Iguazú, provincia de Misiones; Psathyromyia lanei (Barretto & Coutinho) en Posadas, Misiones; y Pintomyia torresi (Le Pont & Desjeux) en Nueva Pompeya, provincia de Chaco. Sciopemyia sordellii (Shannon & Del Ponte) y Brumptomyia brumpti (Larrouse), halladas en Iguazú, son nuevas citas para Misiones.The present paper provides 5 new records of phlebotomine sand flies for Argentina: Micropygomyia oswaldoi (Mangabeira) and Pintomyia bianchigalatiae (Andrade Filho, Aguiar, Dias & Falcão), collected in Iguazú, province of Misiones, Psathyromyia lanei (Barretto & Coutinho), from Posadas, Misiones; and Pintomyia torresi (Le Pont & Desjeux), from Nueva Pompeya, province of Chaco. Sciopemyia sordellii (Shannon & Del Ponte), and Brumptomyia brumpti (Larrouse), collected in Iguazú, are new records for the province of Misiones.

We show the results of application of Differential SAR Interferometry to the MW 7.4, August 17, 1999, Izmit earthquake, Western Turkey. The differential interferogram is obtained using an interferometric ERS2 ascending pair with a time interval of 35 days (August 13th - September 17th). The fringe pattern clearly defines the coseismic displacement field extended in an area of about 100 km N-S and 120 km E-W. The analysis of the interferogram shows the right-lateral strike-slip movement on the activated section of the North Anatolian fault system. The maximum SAR-detected displacement ranges between 117.6 cm and 134.4 cm in the proximity of Gölcük. We invert SAR data for uniform dislocation on a single fault plane using a Montecarlo procedure, with the aim of testing a large set of a priori possible asperity distributions on the fault. We then use a forward modeling approach to evaluate the slip variability for the dislocation using additional constraints as surface offsets and seismicity distribution: in this case we allow unit cells to undergo different values of slip in order to refine the initial dislocation model. Misfits between SAR data and modeled slant range displacements are generally low for all our models (~ 12 cm). Our results indicate that slip is concentrated in the central-western part of the fault, in the upper 10-15 km, tapering to the fault tips. For the Izmit case, we note that a well constrained fault model can be obtained only integrating DInSAR data with additional observations. This is mainly due to an undersampling of the displacement field by DInSAR, caused by decorrelation and lack of image data.

Mangrove trees tend to be larger and mangrove communities more diverse in tropical latitudes, particularly where there is high rainfall. Variation in the structure, growth and productivity of mangrove forests over climatic gradients suggests they are sensitive to variations in climate, but evidence of changes in the structure and growth of mangrove trees in response to climatic variation is scarce. Bomb-pulse radiocarbon dating provides accurate dates of recent wood formation and tree age of tropical and subtropical tree species. Here, we used radiocarbon techniques combined with X-ray densitometry to develop a wood density chronology for the mangrove Avicennia marina in the Exmouth Gulf, Western Australia (WA). We tested whether wood density chronologies of A. marina were sensitive to variation in the Pacific Decadal Oscillation Index, which reflects temperature fluctuations in the Pacific Ocean and is linked to the instrumental rainfall record in north WA. We also determined growth rates in mangrove trees from the Exmouth Gulf, WA. We found that seaward fringing A. marina trees (~10 cm diameter) were 48 ± 1 to 89 ± 23 years old (mean ± 1 σ) and that their growth rates ranged from 4.08 ± 2.36 to 5.30 ± 3.33 mm/yr (mean ± 1 σ). The wood density of our studied mangrove trees decreased with increases in the Pacific Decadal Oscillation Index. Future predicted drying of the region will likely lead to further reductions in wood density and their associated growth rates in mangrove forests in the region.

An increasing incidence of rectal injuries following patient self-induced harmful acts, aimed to sexual or laxatives porpouses, is a fact reported in literature (El-Ashaal et al., 2008). We herein report a case of severe hemoperitoneum related to a middle and upper rectal third seromuscolar tear caused by a self-induced fecal evacuation by means of an arrow with a covered cork tip. An urgent intestinal diversion by means of a Hartmann's operation was performed. The clinical case is presented in relation to the literature debate, regarding the issue of primary repair or resection and anastomosis versus fecal diversion for penetrating rectal injuries (Fabian, 2002; Cleary et al., 2006; Office of the Surgeon General, 1943; Busic et al., 2002). In conclusion, the importance of avoiding an anastomotic breakdown in a patient undergoing a hemorrhagic shock is highlighted.

Mulga, comprised of a complex of closely related Acacia spp., grades from a low open forest to tall shrublands in tropical and sub-tropical arid and semi-arid regions of Australia and experiences warm-to-hot annual temperatures and a pronounced dry season. This short synthesis of current knowledge briefly outlines the causes of the extreme variability in rainfall characteristic of much of central Australia, and then discusses the patterns and drivers of variability in carbon and water fluxes of a central Australian low open Mulga forest. Variation in phenology and the impact of differences in the amount and timing of precipitation on vegetation function are then discussed. We use field observations, with particular emphasis on eddy covariance data, coupled with modelling and remote sensing products to interpret inter-seasonal and inter-annual patterns in the behaviour of this ecosystem. We show that Mulga can vary between periods of near carbon neutrality to periods of being a significant sink or source for carbon, depending on both the amount and timing of rainfall. Further, we demonstrate that Mulga contributed significantly to the 2011 global land sink anomaly, a result ascribed to the exceptional rainfall of 2010/2011. Finally, we compare and contrast the hydraulic traits of three tree species growing close to the Mulga and show how each species uses different combinations of trait strategies (for example, sapwood density, xylem vessel implosion resistance, phenological guild, access to groundwater and Huber value) to co-exist in this semi-arid environment. Understanding the inter-annual variability in functional behaviour of this important arid-zone biome and mechanisms underlying species co-existence will increase our ability to predict trajectories of carbon and water balances for future changing climates.

We compute displacement and stress due to a normal fault by means of two-dimensional plane-strain finite-element analysis. To do so, we apply a system of forces to the fault nodes and develop an iterative algorithm serving to determine the force magnitudes for any slip distribution. As a sample case, we compute the force magnitudes assuming uniform slip on a 10-km two-dimensional normal fault. The numerical model generates displacement and stress fields that compare well with the analytical solution. In fact, we found little difference in displacements (

A geodynamic interpretation of the Northern Marche region is diffi cult, the zone being characterized by complex structures which cannot be defi ned in the form of a simple, standard model. It is unquestionable that the geodynamic setting, whatever it is, bears a strong infl uence on the seismic hazard assessment of a region, and this is the background reason for the present note. In order to obtain a more detailed picture of seismological evidence in this zone, 11 new fault plane solutions of crustal events with 2.9 < M < 4.3 were calculated, using data recorded by the national seismic network of the Istituto Nazionale di Geofi sica e Vulcanologia in the period 1990-2000. The aim is to add local information to the previous studies by Frepoli and Amato (1997, 2000). A possible result of this new study is the division of the Northern Marche region into three areas with different focal mechanism categories: the inner area of the Apenninic belt, the Adriatic on-shore and the Adriatic off-shore. This note is intended to be a contribution to update seismological evidence in the Northern Marche region.

In recent years, the recovery of some historical documents has permitted us to operate the seismographs used by Alessandro Serpieri (1823-1885) at the Observatory of the University of Urbino in the XIX century. The space-time concept of sensor network was already clear to Serpieri and he tried to apply this concept to the analysis of seismic phenomena in Italy. This paper reviews the history of the Urbino Observatory from Serpieri's age to present times. The historical region of Montefeltro, where Urbino is the main town, is affected by seismicity with typical magnitudes between 2.2 and 2.5. Most of these events occur in the upper 15 km of the crust. The seismicity of the neighbouring regions is mainly concentrated in three zones: Northern Rimini, the Apennine belt and the Sibillini Mountain area. From the overall data, it is possible to infer that there is a basin characterised by microseismicity and essentially dominated by a compressive tectonic regime in the Montefeltro area. Furthermore seismological data seem to show a "quiet" segment, separating the extension area from the compression area, characterised by a low concentration of seismic events.

In the Umbria-Marche region, space and time variations concerning the b value were studied by instrumental seismicity from January 1987 to May 1999, according to the Bender method. Data were divided into two partially independent data sets. The first set, (January 1987 - December 1996), does not include the Colfiorito seismic sequence that occurred in the autumn of 1997. The second data set includes all events from January 1987 to May 1999. Using square cell dimensions of 80, 40 and 20 km, the examined area was divided respectively into three grids. The b value was estimated for each cell using the first data set, thus allowing us to reveal b value space variations and determine the resolution. To evaluate the stability of our result we estimated the b value on the basis of historical seismicity within the region. Several synthetic tests were also performed to estimate the stability of the Bender method and to verify its consistency with respect to other methods commonly used. Finally we estimated the b values using the second data set to prove the time variations. Results from the area examined show that the lowest possible spatial resolution of the b value is about 40 km and that there is a correlation between the b value pattern and the main active tectonic structures of the area. The most important time variations occur within the Colfiorito area, in which the b value drops significantly within the second data set. Results suggest two different ways of strain release: the first one produces continuous seismicity that spreads all over the examined area, while the second, which concerns stronger earthquakes, is localized.