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The Campo de Calatrava Volcanic Field (CCVF) located in central-southern Spain (along with Selva-Emporda in Catalonia, NE Spain) is regarded as one of the most important CO emitting zones in Peninsular Spain. Here, we report and evaluate new molecular and isotopic geochemistry of thermal waters and COrich gas discharges from the CCVF. Locally, these CO-rich fluid emissions represent the remnants of the past volcanic activity that affected this area from the late Miocene through the Quaternary, with the most recent events occurring in the Holocene. The locations of discharging fluids and previous volcanic centers appear to be aligned along well-defined NW-SE and NNW-SSE lineaments, with subordinate trends in the ENE-WSW direction. The chemical and isotopic composition of the thermal waters suggests a meteoric origin, dominated by three distinct geochemical facies: 1) HCO-Mg(Ca) type waters, associated with a relatively shallow aquifer and related to the interaction of meteoric waters with CO-rich gases, alkaline volcanic products, and sedimentary formations, 2) SO(Cl)-Ca(Mg) type waters, which stems from the two rivers (Guadiana and Jabalón) that drain Triassic evaporitic rocks before entering the study area, and 3) HCO-Na type waters, hosted in deep geopressurized CO-rich reservoirs within the Ordovician basement rocks. The Sr/Sr isotopic compositions (ranging between 0.70415 and 0.71623) and δS-SO values (+10.7 to +18.3‰ vs. CDT) of CO-rich fluids are consistent with interactions between water and either the Paleozoic basement, Triassic evaporites, Quaternary volcanic rocks, or a combination thereof. Dissolution of a CO-rich gas phase into the aquifer produces low pH values (down to 5.4) and enhances water-rock interactions causing relatively high salinity (Total Ionic Salinity: up to ∼185 meq/L). Carbon dioxide is by far the most abundant gas constituent (up to 992 mmol/mol) and is dominated by mantle-derived sources as indicated by the combination of relatively high helium isotopic ratios (up to 2.7 R/Ra), high isotopic ratios of carbon in CO (ranging between −6.8 and −3.2‰ V-PDB), and the carbon isotopic signature of TDIC (from −6.8 to +2.2‰ vs. VPDB). In the last two decades, numerous (CO-rich) gas blowouts have occurred in the area during well drillings, suggesting the presence of a geopressurized gas reservoir at relatively shallow depth., The Municipality of Almagro is gratefully acknowledged for the help provided during the sampling activities. We would like to thank Dr. Luis Perez del Villar for his help during the first sampling fieldwork at CCVF. We wish to thank D. Melero Cabañas who accompanied us in the field to collect the water samples during the first survey. Many thanks are also due to the personnel of Amphos21 (J. Bruno, A. Cedez, F. Grandia) and Ciudad de la Energia (D. Angel) and F. Capecchiacci (Dept. Earth Science of Florence) for their help during the second survey. We would like to acknowledge the comments and suggestions provided by two reviewers, who greatly improved an early version of the manuscript. This work was partially funded by Ciudad de la Energia (Resp. OV; Grant contract: ALM-08-006) and the Laboratory of Stable Isotopes and Fluid Geochemistry of the Department of Earth Sciences (University of Florence).

In this paper we report an experimental study to assess the process of Sr-isotope uptake from the soil and its transfer to the grapevine and then to the wine made through micro-vinification. The experimental work has been carried out with a deep control of the boundary conditions (i.e., type of soil, geologic substratum, ground water supply, etc.) on 11 selected vine-plant sites over a period of four harvest years. Sr-isotopes have been determined on grape-bunches, grapevine sap, on the bioavailable fraction of the soil, on bulk soil, and on the rocks of the substratum. No significant Sr-isotope variability has been observed among micro-vinifications from different harvest years. A slight but significant Sr-isotope variability occurred among wines from rows embedded on different soil type. The Sr-isotope data on micro-vinifications well match those of grapevine sap and bioavailable fraction of soils, all of them falling well within the whole geological range of the bedrock, despite an evident decoupling between bioavailable fraction, whole soils and bedrocks does exist. This decoupling has been ascribed to differential geochemical behaviour of minerals in response to pedogenetic processes. The findings of our experiments indicate that the biological activity of the vine is not able to change the original 87Sr/86Sr composition up-taken from the bio-available fraction of the soil. Thus, the 87Sr/86Sr of the wine is an unadulterated feature of the terroir.

An analytical protocol for high-precision, in situ microscale isotopic investigations is presented here, which combines the use of a high-performing mechanical microsampling device and high-precision TIMS measurements on micro-Sr samples, allowing for excellent results both in accuracy and precision. The present paper is a detailed methodological description of the whole analytical procedure from sampling to elemental purification and Sr-isotope measurements. The method offers the potential to attain isotope data at the microscale on a wide range of solid materials with the use of minimally invasive sampling. In addition, we present three significant case studies for geological and life sciences, as examples of the various applications of microscale 87Sr/86Sr isotope ratios, concerning (i) the pre-eruptive mechanisms triggering recent eruptions at Nisyros volcano (Greece), (ii) the dynamics involved with the initial magma ascent during Eyjafjallajökull volcano’s (Iceland) 2010 eruption, which are usually related to the precursory signals of the eruption, and (iii) the environmental context of a MIS 3 cave bear, Ursus spelaeus. The studied cases show the robustness of the methods, which can be also be applied in other areas, such as cultural heritage, archaeology, petrology, and forensic sciences.

Odontología

La ricostituzione della filiera produttiva e la tracciabilità dell’intero percorso degli alimenti dalla produzione alla vendita sono divenuti una parte essenziale della ricerca nel campo dell’agroalimentare, soprattutto per gli alimenti di pregio. Tra questi il vino è sicuramente il prodotto che è stato oggetto di maggiore attenzione, anche in considerazione del progressivo sviluppo dei marchi DOC e DOCG e dell’importanza economica assunta dal concetto di Terroir, che ne definisce l’inscindibile legame con il territorio come elemento determinante per la sua valorizzazione economica. La comunità scientifica e gli organismi internazionali hanno dedicato grande attenzione alla messa a punto di metodologie scientifiche e tecnologie idonee alla certificazione di qualità dei vini di pregio. Le analisi dei rapporti isotopici di elementi leggeri (idrogeno, carbonio e ossigeno) sono diffusamente utilizzate per il controllo dell’autenticità del vino e sono state inserite in regolamenti CEE e nazionali. Quest’analisi non è tuttavia risolutiva per il riconoscimento e la tracciabilità geografica del vino a causa della loro forte dipendenza da fattori climatici locali e dall’andamento atmosferico stagionale. Per questo motivo la ricerca si è indirizzata verso lo studio di elementi caratteristici del sistema substrato/suolo di provenienza con l’obiettivo di risalire dalla composizione chimico-fisica del prodotto vino al suo areale (territorio) d’origine. Infatti la composizione chimica e isotopica della matrice inorganica è legata alla natura del suolo di impianto della vigna, che eredita le proprie caratteristiche composizionali e qualitative direttamente dal substrato geologico e dunque può rappresentare la sua impronta digitale e fornire uno strumento efficace per la sua tracciabilità.

The Roccamonfina Volcano is characterised by a two stage volcanic activity, divided by volcano- tectonic collapses. Ultrapotassic leucite-bearing rocks are confined in the pre-caldera stage and display geochemical characteristics very similar to those of other volcanoes of the Roman Province. Shoshonitic rocks have been poured out from cinder cones and domes both within the caldera and on the external flanks of the pre-caldera Roccamonfina volcano. On the basis of new trace element and Sr-Nd-Pb isotope data, Roccamonfina shoshonitic rocks show differences with respect to shoshonites from Northern Roman Province, and recall closely those of the Neapolitan volcanoes. Eventually, in the very last phases of volcanic activity, sub-alkaline magmas are found both as enclaves in trachytic domes, and as final lavas vented by a fracture within the Monte Santa Croce dome. Ultrapotassic rocks that built up the pre-caldera composite volcano are plagioclase-bearing leucitites and, similarly to other Roman Provinces volcanoes, have by high levels of incompatible trace elements with an orogenic signature having troughs at Ba, Ta, Nb, Ta, and Ti, and peaks at Cs, K, Th, U, and Pb. Initial values of 87Sr/86Sr range from 0.709261 to 0.709987, of 143Nd/144Nd range from 0.512134 to 512195, of 206Pb/204Pb range from 18.788 to 18.851, of 207Pb/204Pb range from 15.685 to 15.701, and of 208Pb/204Pb range from 39.048 to 39.076. Shoshonites show a larger differentiation degree but, as a whole, lower levels of incompatible trace elements with similar pattern of incompatible trace element contents with respect to earlier ultrapotassic leucite-bearing rocks. On the other hand, shoshonitic rocks have Sr, Nd, and Pb isotopes consistently different than pre-caldera ultrapotassic leucite-bearing rocks. 87Sr/86Sr ranges from 0.706604 to 0.708332, 143Nd/144Nd ranges from 0.512179 to 0.512382, 206Pb/204Pb ranges from 18.838 to 19.153, 207Pb/204Pb ranges from 15.661 to 15.694, and 208Pb/204Pb ranges from 38.999 to 39.212. High-K calc-alkaline samples have intermediate isotopic values between ultrapotassic plagioclase leucitites and shoshonites, but the lowest levels of incompatible trace element contents. The genesis of this uncommon association is investigated in details. It is argued that ultrapotassic magmas have been generated in a modified lithospheric mantle after crustal- derived metasomatism. Interaction between metasomatic agent and lithospheric upper mantle produced a low-melting metasomatised veined network. Pure partial melting of vein mineralogy produced pre-caldera leucite-bearing ultrapotassic magmas. Leucite-free magmas have been generated further melting, at higher temperature, of the same meltasomatic mantle previously depleted by the ectraction of ultrapotassic rocks. We suggest that a key role in this second episode of melting is played by little but significant addition of astenospheric upper mantle material from foreland mantle, flowing through the corner of the Ionian present day subducted slab during its roll-back

Osservare i territori in contrazione attraverso la pratica del cammino, permette di esperire le sfumature che caratterizzano questi territori posti ai margini: in questo contributo si osserva la dimensione dell'inselvatichimento. Che si tratti di aree urbane dismesse, di aree rurali non più coltivate o di aree montane troppo complesse per rispondere alla competitività produttiva di territori più accessibili, nel momento in cui l'essere umano rallenta i processi di continua trasformazione del suolo e dei significati, il selvatico si riappropria degli spazi dimenticati. Si propone così uno sguardo sulle diverse realtà in cui il selvatico ri-appare e si prova a considerarlo come un nuovo soggetto possibile in quello che è stato chiamato "il secolo urbano". Observing the shrinking territories through the practice of walking allows us to experience the nuances that characterize territories placed on the margins: in this contribution I observe the dimension of re-wilding. Whether it is dismissed urban areas, rural areas no longer cultivated or mountain areas too complex to respond to the productive competitiveness of more accessible territories, when human beings slow down the processes of continuous transformation of the soil, the wild takes back the forgotten spaces. Thus I propose to observe the different realities in which the re-wilding processes appear and I try to consider the wild as a new possible subject in what has been called "the urban century".

Il saggio riflette sul tema della Temporaneità considerata, ormai, prassi strettamente connessa a quello dell’urban shrinkage, condizione centrale nel dibattito scientifico quanto nell’agenda politica di diverse città europee. Dagli anni ’90 a livello globale, nelle città e nei territori abbondano i luoghi dell’abbandono e dell’inutilizzato che ci restituiscono uno scenario di progressiva contrazione non solo demografica ma anche economica e sociale. In questo controverso quadro di mutazione strutturale delle città e dei territori si stanno sperimentando strategie, strumenti e schemi di governance efficaci per governare la decrescita e l’abbandono il cui comun denominatore è il ricorso all’uso temporaneo. Tale modalità d’uso dello spazio è diventato un vero e proprio trend urbano rivestendo, contestualmente, sempre più centralità tanto nel dibattito scientifico quanto in quello politico, traducendosi in innumerevoli pratiche che stanno ridisegnando i paesaggi dell’abbandono. Tali esperienze possono essere considerate occasioni per ridefinire gli immaginari urbani? O per sperimentare nuove forme di produzione di pubblico e restituire centralità alle comunità locali nei processi di riconversione e trasformazione urbana? Il saggio si propone di dare una risposta a tali quesiti. The essay reflects on the theme of Temporariness considered a practice closely related to that of urban shrinkage, a central condition to the scientific debate and into the political agenda of several European cities. Since the 1990s globally, places of abandonment have abounded in cities, returning us to a scenario of progressive shrinkage not only demographic but also economic and social. In this controversial framework of structural mutation of cities and territories effective strategies, tools and governance schemes are being experimented with to govern degrowth and abandonment whose common denominator is the recourse to temporary use. This mode of space use has become a real urban trend, contextually holding increasing centrality in both scientific and political debate, translating into countless practices that are reshaping the landscapes of abandonment. Can such experiences be considered opportunities to redefine urban imaginaries? Or to experiment with new forms of public production and to restore centrality to local communities in processes of urban redevelopment and transformation? The essay aims to provide an answer to these questions.

Partendo da una difficoltà che è quella di dare un nome a dimensioni che non hanno chiara definizione, il testo introduce il lavoro di indagine e di progetto sviluppato durante e in seguito al Laboratorio del Cammino 2022. In questa occasione si sono attraversati i territori in contrazione delle province di Biella e Torino e lo sguardo è stato rivolto ai/alle pionieri/e (umani e non) capaci di abitare un territorio alle volte ostile ma ricco di nuove possibilità. Starting from the effort needed to name realities that have no clear definition, the contribution introduces the field research and project work developed during and after the "Laboratorio del Cammino", 2022. On this occasion, territories in contraction were crossed in the provinces of Biella and Turin and the gaze was turned to the pioneers (human and not-human) capable of inhabiting a territory that is sometimes hostile but full of new possibilities.

Partendo dalla concezione di "rischio" e contrapponendola a quella di "azzardo", il testo introduce il lavoro di indagine e le proposte interattive nate durante e in seguito dell'esperienza del Laboratorio del cammino che nel 2021 attraversavai territori in contrazione delle province di Torino e Biella. Starting from the concept of "risk" and opposing it with that of "hazard", the text introduces the investigation work and the interactive proposals developed during and following the experience of the "Laboratorio del Cammino" which in 2021 crossed the shrinking territories of the Turin and Biella provinces.

(1) Background: It is recommended that an athlete, in order to ensure correct nutrition and performance, should consume between 1.2 and 2.0 g/kg/day of protein, while the daily recommended protein intake for a non-athlete is 0.8and 0.9 mg/kg/day. It is unclear if athletes living in Mediterranean countries are able to meet protein requirements without supplementation, since Mediterranean diet de-emphasizes meat and meat products. (2) Methods: 166 athletes (125 males) enrolled between 2017 and 2019 were required to keep a dietary journal for three consecutive days (2 workdays and 1 weekend day). Athletes had to be &gt, 18 years old, train in a particular sport activity more than 3 h a week and compete at least at an amateur level. Journal data were collected and then translated into macro-nutrient content (grams of protein, carbohydrates, and lipids) by a nutritionist. (3) Results: The protein intake reported by this specific population vary slightly from the Academy of Nutrition and Dietetics (AND), Dietitians of Canada (DC), and the American College of Sports Medicine (ACSM) joint statement recommendation level. Average protein levels without protein supplementation fell within the protein guidelines. Counterintuitively, the intake among those who supplemented their diet with protein was higher compared with those who did not, even when excluding the contribution of supplements. Although the majority of subjects participating in the study were able to meet protein intake recommended for athletes without protein supplementation, 27% of athletes were below the guideline range. (4) Conclusions: these data suggest that athletes&rsquo, nutrition should be more often evaluated by a nutritionist and that they will benefit from increasing their nutritional knowledge in order to make better food choices, resorting to protein supplementation only when effectively needed.

In evolutionary terms, life on the planet has taken the form of independently living cells for the majority of time. In comparison, the mammalian radiation is a relatively recent event. The common mammalian ancestor was probably small and short-lived. The "recent" acquisition of an extended longevity and large body mass of some species of mammals present on the earth today suggests the possibility that similar cellular mechanisms have been influenced by the forces of natural selection to create a convergent evolution of longevity. Many cellular mechanisms are potentially relevant for extending longevity; in this assay, we review the literature focusing primarily on two cellular features: (1) the capacity for extensive cellular proliferation of differentiated cells, while maintaining genome stability; and (2) the capacity to detect DNA damage. We have observed that longevity and body mass are both positively linked to these cellular mechanisms and then used statistical tools to evaluate their relative importance. Our analysis suggest that the capacity for extensive cellular proliferation while maintaining sufficient genome stability, correlates to species body mass while the capacity to correctly identify the presence of DNA damage seems more an attribute of long-lived species. Finally, our data are in support of the idea that a slower development, allowing for better DNA damage detection and handling, should associate with longer life span.

A huge progress has been achieved in mammalian in vitro technique. Instead, of the many trials to develop marine invertebrate cell cultures, only a few have obtained them and only from few tissues. Since in vitro cell culture of invertebrates could be very useful for many aspects of basic and applied science, in this work we investigate and describe the development of a technique for the establishment of cell cultures from gill, mantle and gonadic tissue of the Manila clam (Ruditapes philippinarum). We maintained viable cultures for up to 25 days. Culture viability and proliferation were tested with 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) and with trypan blue, while an antibody against the ATP-dependent RNA helicase VASA, a protein expressed in the germline, and in multipotent stem cells of some animals, was used to verify the presence of these cell types. Following the described protocol: 1) explant resulted the better source to obtain cell cultures, when compared to enzymatic dissociation; 2) cultures of suspended cells were viable for longer period than adherent cells; 3) cell cultures obtained from tissues sampled in September-October performed better compared to other periods of the year, regarding maintenance and growth; 4) the tissue from which we obtained longer-lived cell cultures was gonadic tissue, especially form samples that show more undifferentiated germ cells and more VASA-stained cells. This study describes the challenges concerning the development of in vitro culture techniques for aquatic invertebrates.

A large number of basic researches and observational studies suggested the cancer preventive activity of vitamin E, but large-scale human intervention trials have yielded disappointing results and actually showed a higher incidence of prostate cancer although the mechanisms underlying the increased risk remain largely unknown. Here we show through in vitro and in vivo studies that vitamin E produces a marked inductive effect on carcinogen-bioactivating enzymes and a pro-oxidant status promoting both DNA damage and cell transformation frequency. First, we found that vitamin E in the human prostate epithelial RWPE-1 cell line has the remarkable ability to upregulate the expression of various phase-I activating cytochrome P450 (CYP) enzymes, including activators of polycyclic aromatic hydrocarbons (PAHs), giving rise to supraphysiological levels of reactive oxygen species. Furthermore, our rat model confirmed that vitamin E in the prostate has a powerful booster effect on CYP enzymes associated with the generation of oxidative stress, thereby favoring lipid-derived electrophile spread that covalently modifies proteins. We show that vitamin E not only causes DNA damage but also promotes cell transformation frequency induced by the PAH-prototype benzo[a]pyrene. Our findings might explain why dietary supplementation with vitamin E increases the prostate cancer risk among healthy men.

Across species, development and longevity are tightly linked. We discuss the relevant literature and suggest that the root for this stringent relationship is the rate of development. The basis for the relationship between rate of development and longevity lies in adaptations that have occurred through evolution at multiple levels of biological complexity: organism, organ, cellular, and molecular. Thus, the analysis of the relationship is of interest for multiple fields of biology.

The geologic map of the Neogene Cabo de Gata volcanic zone is presented together with a comprehensive volcanic stratigraphy and structure based on logging, correlation and mapping. Volcanic rocks are interbedded with sedimentary rocks throughout the Cabo de Gata volcanic zone. The volcano-sedimentary succession of Cabo de Gata has been divided into formations according to lithology, age, composition and stratigraphic position. The contacts between sedimentary units and volcanic units and between formations are unconformities. Sedimentary units were deposited during periods of volcanic repose. The depositional environment of volcanism in Cabo de Gata is characterized as shallow-water submarine to emergent based on lithofacies of volcanic rocks and on fossil content and sedimentary structures of sedimentary rocks. The eruptive style in Cabo de Gata is dominantly effusive, although small-volume explosive eruptions due to magma-water interaction processes and to explosions of lava flow and domes complexes occurred.

X-linked hypophosphatemia (XLH) is a genetic disorder related to alterations in bones and teeth formation, due to low levels of phosphate in blood. Oral findings in XLH have been enamel and dentine abnormalities, high pulp horns, large pulp chambers, and some cases of periapical abscesses related to teeth without caries or traumatic injuries. The aim of our study was to assess the presence of enamel alterations, such as microclefts and/or structure defects in patients with XLH and give guidelines of prevention of XLH dental complications. History taking, oral clinical and radiological examination in 10 young patients affected by XLH (average age of 9) and in 6 patients without XLH (average age of 8). Impressions were performed on the vestibular surfaces of teeth in order to obtain replicas. The replicas were analyzed using scanning electron microscope (SEM) and compared to replicas of control group. The images of replicas of XLH patients showed deep microclefts and irregular enamel surface structure compared to replicas of control group. The replica of a patient with spontaneous periapical abscesses showed numerous enamel crater-shaped depressions and deep microcleavages penetrating into the enamel thickness. In absence of caries or fractures, the abscesses pathogenesis may be related to microcleavages of the enamel and dentin, which allow bacterial invasion of the pulp. There could be a relationship between XLH disease and enamel abnormalities.

""\\"The radiogenic isotopic compositions of inorganic heavy elements such as Sr, Nd, and Pb of the food chain may constitute a reliable geographic fingerprint, their isotopic ratios being inherited by the geological substratum of the territory of production. The Sr isotope composition of geomaterials (i.e., rocks and soils) is largely variable, and it depends upon the age of the rocks and their nature (e.g., genesis, composition). In this study we developed a high-precision analytical procedure for determining Sr isotopes in wines at comparable uncertainty levels of geological data. With the aim of verifying the possibility of using Sr isotope in wine as a reliable tracer for geographic provenance, we performed Sr isotope analyses of 45 bottled wines from four different geographical localities of the Italian peninsula. Their Sr isotope composition has been compared with that of rocks from the substrata (i.e., rocks) of their vineyards. In addition wines from the same winemaker but different vintage years have been analyzed to verify the constancy with time of the Sr-87\\\\\\\/Sr-86. Sr isotope compositions have been determined by solid source thermal ionization mass spectrometry following purification of Sr in a clean laboratory. Sr-87\\\\\\\/Sr-68 of the analyzed wines is correlated with the isotopic values of the geological substratum of the vineyards, showing little or no variation within the same vineyard and among different vintages. Large Sr-87\\\\\\\/Sr-86 variation is observed among wines from the different geographical areas, reinforcing the link with the geological substratum of the production territory. This makes Sr isotopes a robust geochernical tool for tracing the geographic authenticity and provenance of wine.\\"""

Objective: To evaluate whether the debonding procedure leads to restitutio ad integrum of the enamel surface by investigating the presence of enamel within the bracket base remnants after debonding. Materials and Methods: Sixty patients who completed orthodontic treatment with fixed appliances were included. A total of 1068 brackets were microphotographed; the brackets presenting some remnants on the base (n = 818) were selected and analyzed with ImageJ software to measure the remnant area. From this population a statistically significant sample (n = 100) was observed under a scanning electron microscope to check for the presence of enamel within the remnants. Energy dispersive x-ray spectrometry was also performed to obtain quantitative data. Results: Statistically significant differences in the remnant percentage between arches were observed for incisor and canine brackets (P < .0001 and P = .022, respectively). From a morphologic analysis of the scanning electron micrographs the bracket bases were categorized in 3 groups: group A, bases presenting a thin enamel coat (83%); group B, bases showing sizable enamel fragments (7%); group C, bases with no morphologic evidence of enamel presence (10%). Calcium presence was noted on all evaluated brackets under energy dispersive x-ray spectrometry. No significant difference was observed in the Ca/Si ratio between group A (16.21%) and group B (18.77%), whereas the Ca/Si ratio in group C (5.40%) was significantly lower than that of the other groups (P < .323 and P = .0001, respectively). Conclusion: The objective of an atraumatic debonding is not achieved yet; in some cases the damage could be clinically relevant.

Scanning electron microscope evaluation could be criticized if the method adopted to correct for bias is not specified in the study design. Observers can draw conclusions from images unconsciously chosen to best support their research hypotheses, impairing the basic research principle of operator's impartiality. In this study, a systematic observation method has been described and verified for repeatability. The number and the observation points on a certain specimen have been predetermined using a scheme along with observation rules previously established in the research protocol. When our instrument is used at an operating magnification between 500x and 1,000x (corresponding to a frame of 250x190 micro and 120x90 micro, respectively), the method allowed 100% repeatable observation frames, with linear frame errors in finding an observation point of 12.5% in length and 16.8% in height. With modifications to accommodate research objective and statistical requirements, the method could be applied to many SEM observation study.