Application of Synchrotron Radiation X-Ray Fluorescence (μ-SRXF) and X-Ray Microanalysis (SEM/EDX) for the Quantitative and Qualitative Evaluation of Trace Element Accumulation in Woody Plants

Different analytical techniques were applied to describe the localization of lead and chromium in the tissues of walnut (Juglans regia) and maple (Acer saccharinum) plants exposed to soils that had been artificially contaminated with heavy metals. Two X-ray-based techniques, synchrotron radiation X-ray fluorescence (μ-SRXF) and X-ray microanalysis (SEM/EDX), were utilized in association with induced coupled plasma optical emission spectrometry (ICP-OES). These techniques allowed the definition of maps showing a preferential accumulation of lead in the root periderm of both plants and a limited translocation of the metal to the stems. Accordingly, estimation of the lead concentration with ICP-OES showed that roots contained, on a dry weight basis, 20 to 40 times more metal than stems. Chromium, supplied to the plants as Cr³⁺ or Cr⁶⁺, was taken up only in the latter case and in limited amounts. In general, walnut was more efficient than maple in the uptake of lead and chromium. The combination of X-ray-based techniques and ICP-OES proved to be a powerful quantitative tool in mapping, with high definition, the sites of metal storage in plant tissues.     

A preliminary study in the use of RAPD markers in detecting genetic differences in hatching patterns of Chirocephalus diaphanus Prévost, 1803 (Crustacea: Anostraca)

Randomly amplified polymorphic DNA (RAPD) was used to detect intra-clutch genetic differences of the anostracan Chirocephalus diaphanus Prévost, 1803. Thirty primers were tested on 130 nauplii from three different clutches. Two primers produced five repeatable and well-defined amplification products, that were used as genotype markers. Nauplii hatched after three successive dehydration and rehydration cycles. Consistent genetic differentiation was observed among nauplii from the same clutch. Significant association between presence or absence of three polymorphic amplification products and hatching after different stimuli was observed. These results suggest that RAPD markers might be related to the difference in nauplii hatching and provide reasonable support to the hypothesis that the source of the observed variation in hatching phenology are genetic as well as epigenetic.     

War in Ukraine – Changing the Polish paediatric population as a consequence of forced migration (a preliminary study)

The study aimed to briefly analyse the basic anthropometric characteristics of Ukrainian adolescents against their Polish counterparts. The study was school-based, carried out April–June 2022 and followed a cross-sectional model. It included 642 children from Poland and Ukraine (aged 10–15) who attended ten randomly selected primary schools in Kraków (Poland). Noticeable differences between Polish and Ukrainian adolescents were observed. As the analysed characteristics included features such as the amount and distribution of body fat, they can be crucial in the indirect assessment of health status, particularly the risk of cardiovascular and metabolic diseases. This in turn, can help to properly accommodate the changing needs of the population.     

A preliminary study on the distribution patterns of endemic species of Fulgoromorpha (Hemiptera,?Auchenorrhyncha) in Iran

Iran is known as the most complex and varied country in southwest Asia, in terms of geography, vegetation, climate and consequently biological diversity. The rather high number of recorded endemic species of Fulgoromorpha in Iran indicates a high potential for speciation in some areas.In this study, in order to identify the endemic zones for Fulgoromorpha of Iran, three main biogeographic regions of the country were divided into 13 primary zones, mainly according to the distribution of published and unpublished locality records of endemic species. Using Venn diagrams and cluster analyses on the primary zones, 6 final endemic zones were recognized: Caspian zone, southern slopes of Alborz, Zagros Mountains, Kerman Mountains, Khorasan Mountains, and Baluchestan and Persian Gulf coasts. Then a similarity map was produced for endemic zones using a Multidimensional analysis (Alscal) and the differences between the positions of the same zones in the similarity and geographic maps were discussed.     

Pollen content study of the lower atmosphere in León (Spain) by use of a tethered balloon

This study was undertaken in order to understand thebehaviour of airborne pollen grains, namely therelationship between their concentrations andconcomitant meteorological parameters, at differentaltitudes (ground level, 200, 400 and 600 m) of thelower atmosphere and its daily variations over onesite (the experimental farm of the University ofLeón). The experimental design involved a tetheredballoon (2.25 m³), an ADAS remote meteorologicalstation and an original radio controlled three headedpollen and spore sampler (called GABIS) using theRotorod design. Each head sampled a differentaltitude, while two control samples were taken atground level. Twenty-one takeoffs could be realized inthe 5 weeks period between end of May and end of June1997. Sampling was done early in the morning and sampling time at each altitudewas of 15 minutes. Results show that 45 differenttypes of pollen grains could be collected at this timeof the year and that significant variations could beobserved in the behaviour of the pollen cloud on a dayto day basis, probably because of differentmeteorological situations. Contrary to what isgenerally believed, pollen was in most cases moreabundant at higher altitude – on average by 30% ascompared to ground level – making evident anaerobiological layer of transport at about 500 m aboveground. This was especially the case for trees (Quercus and Castanea). The atmosphere MixingRatio was the most explicative factor at 200 m, whiletemperature dominated significance analysis at both400 and 600 m.     

Pollen harvest by solitary bees (Ptilothrix relata,Hym. Apidae,Emphorini) in the Argentine pampas – preliminary results

In order to determine the plants foraged by solitary Ptilothrix relata, the pollen content of food provisions was identified. The main pollen types confirm foraging by these bees on Malvaceae, Asteraceae and Onagraceae. Pollen sources belonging to Cucurbitaceae, Dipsacaceae and Portulacaceae families enrich the harvesting range of P. relata. According to the preliminary results, these bees can be considered more as narrowly polylectic than oligolectic because they collect pollen from a few unrelated species. Among the most abundant pollen collected, two major morphological types were recognized: (1) pollen with long spines (4–25?μm), many single apertures and large size (115–135?μm of diameter); and (2) tricolporate pollen with viscin threads.     

Somatic embryo mediated mass production of Catharanthus roseus in culture vessel (bioreactor) – A comparative study

The purpose of this study was to evaluate and compare the use of liquid and solid Murashige and Skoog (MS) medium in different culture vessels for mass production of Catharanthus roseus, an important source of anticancerous compounds, vincristine and vinblastine. Three media conditions i.e. agar-solidified medium (S), liquid medium in agitated conical flask (L) and growtek bioreactor (B) were used. Rapid propagation was achieved through in vitro somatic embryogenesis pathway. The process of embryogenesis has been categorized into induction, proliferation, maturation and germination stages. All in vitro embryogenesis stages were conducted by withdrawing spent liquid medium and by adding fresh MS medium. In optimized 4.52 μM 2,4-D added MS, the callus biomass growth was low in solid (1.65 g) compared to liquid medium in agitated conical flask (1.95 g) and in bioreactor (2.11 g). The number of normal somatic embryos was more in solid medium (99.75/50 mg of callus mass) compared to liquid medium used in conical flask (83.25/callus mass) and growtek bioreactor (84.88/callus mass). The in vitro raised embryos maturated in GA₃ (2.60 μM) added medium; and in bioreactor the embryo growth was high, a maximum length of 9.82 mm was observed at the end of four weeks. These embryos germinated into seedlings in BAP (2.22 μM) added medium and the embryo germination ability was more (59.41%) in bioreactor compared to liquid medium in conical flask (55.5%). Shoot length (11.25 mm) was also high in bioreactor compared to agitated conical flask. The liquid medium used in agitated conical flask and bioreactor increased seedling production efficiency, at the same time it also reduced plant recovery time. The embryo generated plants grew normally in outdoor conditions. The exploitation of medium to large culture vessel or bioreactor may make the process more efficient in getting large number of Catharanthus plant as it is the only source of anti-cancerous alkaloids, vincristine and vinblastine.Abbreviations: BA, N6-benzyladenine; 2,4-D, 2,4-Dichlorophenoxyacetic acid; GA₃, gibberellic acid; NAA, naphthalene acetic acid; MS, Murashige and Skoog (1962) medium; S, agar-solidified medium; L, liquid medium in agitated conical flask; B, growtek bioreactor     

Azadirachtin(A) distinctively modulates subdomain 2 of actin – novel mechanism to induce depolymerization revealed by molecular dynamics study

Azadirachtin(A) (AZA), a potential insecticide from neem, binds to actin and induces depolymerization in Drosophila. AZA binds to the pocket same as that of Latrunculin A (LAT), but LAT inhibits actin polymerization by stiffening the actin structure and affects the ADP–ATP exchange. The mechanism by which AZA induces actin depolymerization is not clearly understood. Therefore, different computational experiments were conducted to delineate the precise mechanism of AZA-induced actin depolymerization. Molecular dynamics studies showed that AZA strongly interacted with subdomain 2 and destabilized the interactions between subdomain 2 of one actin and subdomains 1 and 4 of the adjacent actin, causing the separation of actin subunits. The separation was observed between subdomain 3 of subunit n and subdomain 4 of subunit n + 2. However, the specific triggering point for the separation of the subunits was the destabilization of direct interactions between subdomain 2 of subunit n (Arg39, Val45, Gly46 and Arg62) and subdomain 4 of subunit n + 2 (Asp286, Ile287, Asp288, Ile289, Asp244 and Lys291). These results reveal a unique mechanism of an actin filament modulator that induces depolymerization. This mechanism of AZA can be used to design similar molecules against mammalian actins for cancer therapy.     

Does reproductive success increase with age or with size in species with indeterminate growth? A case study using sand lizards (Lacerta agilis)

Most data on determinants of reproductive success (RS) and reproductive tactics are correlational in nature, and hence cannot be used to infer causation. Consistent patterns-such as an increase in RS with age, as seen in many types of organisms-may result from diverse underlying mechanisms. Ontogenetic increases in RS in mammals and birds may be due largely to direct effects of age (via learning, etc.) but our analyses show that apparently analogous ontogenetic shifts in reproductive tactics and increases in RS in sand lizards (Lacerta agilis) are actually due to ontogenetic changes in body size. When size effects are removed, age exerts very little effect on either reproductive behaviour or RS in either sex. In many taxa, both age and body size may exert important effects on reproductive biology, and disentangling these effects should be a focus of further research.     

Grassland Leafhoppers (Hemiptera: Auchenorrhyncha) as Indicators of Habitat Condition – A Comparison of between-site and between-year differences in Assemblage Composition

The Auchenorrhyncha (leafhoppers) show great potential as indicators of grassland habitat quality, which would make them useful as a conservation tool. However, they are known to have labile populations. The relative importance of site identity and the year of sampling in the composition of leafhopper assemblages on chalk grassland are assessed for two sets of sites sampled twice. The study included a total of 95 sites (one set of 54, the other of 41), and demonstrated that for both sets the vegetation community and geographical location had high explanatory value, while the influence of year was small. The conclusion is that, notwithstanding population fluctuations, the leafhopper assemblages are a good indicator of habitat quality, and represent a potentially valuable tool in grassland conservation and restoration.     

Arabidopsis arenosa (Brassicaceae) from a lead–zinc waste heap in southern Poland – a plant with high tolerance to heavy metals

The morphological traits and tolerance to heavy metals (zinc, cadmium and lead) of two populations of Arabidopsis arenosa (Brassicaceae) were compared. One population was from a zinc–lead waste heap in Bolesław near Olkusz (southern Poland), the other one from the Kampinoski National Park (central Poland). Biometric measurements were done in the field and repeated after cultivation under controlled conditions (garden soil, phytotron chamber). Significant heritable morphological differences between the two populations were found. The plants from the waste-heap are smaller in comparison with the reference population, and their leaves are narrower, thicker with fewer trichomes, indicating. genetic adaptation to xerothermic conditions. The level of tolerance to heavy metals (zinc, cadmium, and lead) was compared by the root test. Very high tolerance to the three metals tested was found in the waste-heap population. Its tolerance exceeded that of four other predominant plant species populations growing on the same waste heap that had previously been tested in our laboratory. We consider the waste-heap population of A. arenosa to be a very suitable ecotype for the study of heavy metal tolerance mechanisms in plants. Responsible Editor: SE: Henk Schat.     

Nutrient fluxes in intertidal communities of a South European lagoon (Ria Formosa) – similarities and differences with a northern Wadden Sea bay (Sylt-Rømø Bay)

During an annual cycle, flux rates of oxygen, nitrate, nitrite, ammonium, phosphate and silicate were measured in light and dark bell jars at three sites in Ria Formosa (Algarve, Portugal) enclosing either a natural macrophytic community (macroalgae on sand or mud, a seagrass bed of Zostera noltii) or bare sediments. The results are compared with a preceeding study in which the same bell jar technique has been applied in the Sylt-Rømø Bay of the northern Wadden Sea. Nitrate flux was mainly directed from the water column to the benthic communities in Ria Formosa, as well as in the Sylt-Rømø Bay. However, nitrate uptake was higher in the northern, more eutrophic study area. In Ria Formosa, nutrient concentrations were lower than in the Sylt-Rømø Bay possibly due to strong water exchange with Atlantic waters. High temperatures and strong insolation had a greater impact on nitrate fluxes in Ria Formosa than in the Sylt-Rømø Bay. Bioturbating macrofauna increased ammonium efflux in the Sylt- Rømø Bay while this effect was not as pronounced in the Ria Formosa study sites. Benthic phosphate uptake dominated in the Ria Formosa and was correlated to initial phosphate concentrations in incoming waters. At both study sites, oxygen and nutrient fluxes were correlated with temperature. Additionally, flux rates were strongly influenced by biotic components and levels of eutrophication. A literature survey showed that mainly in temperate regions, material fluxes increase with temperature, whereas in warmer areas, ammonium and phosphate fluxes between sediment and water were generally lower.     

Accurate Determination of S-Phase Fraction in Proliferative Cells by Dual Fluorescence and Peroxidase Immunohistochemistry with 5-Bromo-2��-Deoxyuridine (BrdU) and Ki67 Antibodies

To ensure the maintenance of tissues in mammals, cell loss must be balanced with cell production, the proliferative activity being different from tissue to tissue. In this article, the authors propose a new method for the quantification of the proliferative activity, defined as the S-phase fraction of actively cycling cells, by dual labeling with fluorescence and peroxidase immunohistochemistry using BrdU (marker of S-phase) and Ki67 antibodies (marker of G₁-, S-, G₂-, and M-phases) after a one-step antigen retrieval. In the generative cell zones of fundic and pyloric glandular stomachs, where the majority of cells were cycling, the authors measured a proliferative activity of 31%. In the epithelium of the forestomach and the skin, where cycling cells are intermingled with G₀ and differentiated cells, proliferative activities were 21% and 13%, respectively. In the adrenal cortex, in which cycling cells were sparsely distributed, the proliferative activity reached 32%. During the regenerative process in the skin after a lesion, the proliferative activity increased in proximity to the wound. The present one-step dual-labeling method has revealed that the proliferative activity is different between tissues and depends on the physiological or pathological state.     

A Phase I,Dose-Finding Study of Sorafenib in Combination with Gemcitabine and Radiation Therapy in Patients with Unresectable Pancreatic Adenocarcinoma: A Grupo Espa?ol Multidisciplinario en Cáncer Digestivo (GEMCAD) Study

Purpose

Sorafenib, an oral inhibitor of B-raf, VEGFR2, and PDGFR2-beta, acts against pancreatic cancer in preclinical models. Due to the radio-sensitization activity of both sorafenib and gemcitabine, we designed a multicenter, phase I trial to evaluate the safety profile and the recommended dose of this combination used with concomitant radiation therapy.

Methods

Patients with biopsy-proven, unresectable pancreatic adenocarcinoma (based on vascular invasion detected by computed tomography) were treated with gemcitabine (300 mg/m2 i.v. weekly ×5 weeks) concurrently with radiation therapy (45 Gy in 25 fractions) and sorafenib (escalated doses in a 3+3 design, from 200 to 800 mg/day). Radiation portals included the primary tumor but not the regional lymph nodes. Patients with planning target volumes (PTV) over 500 cc were excluded. Cases not progressing during chemoradiation were allowed to continue with sorafenib until disease progression.

Results

Twelve patients were included. Three patients received 200 mg/day, 6 received 400 mg/day, and 3 received 800 mg/day; PTVs ranged from 105 to 500 cc. No dose-limiting toxicities occurred. The most common grade 2 toxicities were fatigue, neutropenia, nausea, and raised serum transaminases. Treatment was discontinued in one patient because of a reversible posterior leukoencephalopathy. There were no treatment-related deaths.

Conclusion

The addition of sorafenib to concurrent gemcitabine and radiation therapy showed a favorable safety profile in unresectable pancreatic adenocarcinoma. A dose of 800 mg/day is recommended for phase II evaluation.

Trial Registration

EudraCT 2007-003211-31 ClinicalTrials.gov 00789763     

A 31P1H NMR study of the reactions of [Rh2(μ-Cl)2(cod)2] with unsymmetrical,bidentate ligands and hydrogen

[Rh₂(μ-Cl)₂(cod)₂] reacts with Ph₂PCH₂CH₂OMe (PC₂O), Ph₂P(CH₂)₃NMe₂ (PC₃N), PBuⁿPh₂ or PPh₃ to give [Rh(cod)L₂]Cl (L = PC₂O, PC₃N, PBuⁿPh₂, PPh₃). In the presence of hydrogen, [Rh(cod)L₂]Cl is converted to [RhClH₂L₃]. In contrast, [Rh(cod)(PC₂O)₂]BPh₄ reacts with H₂ to give [RhH₂(PC₂O)₂S₂]BPh₄ (S = solvent). With Ph₂PCH₂CH₂NMe₂ (PC₂N) or Ph₂PCH₂CH₂SMe (PC₂S), [Rh₂(μ-Cl)₂(cod)₂] reacts to form the chelate complexes cis- [Rh(PC₂N)₂]⁺ or cis-[Rh(PC₂S)₂]⁺, neither of which reacts with hydrogen under ambient conditions. The products of the reactions are characterized in situ by ³¹P¹H NMR spectroscopy.     

Is there a relation between genetic susceptibility with cancer? A study about paraoxanase (PON1) enzyme activity in breast cancer cases

Human serum paraoxonase 1 (PON1, EC 3.1.1.2) is a high density lipoprotein (HDL)-associated antioxidant enzyme that not only decreases oxidative stress, but it is also implicated in development of many cancers. Genetic information provides a means of identifying people who have an increased risk of cancer, thus this knowledge of cancer genetics helps to identify the ability to characterize malignancies leading to the development of new therapeutic approaches. Because of this reason, in this preliminary study we aimed to investigate the role of human serum PON1 enzyme activity and phenotypic distribution in 32 breast cancer (BC) patients (age range 28–82) and 35 cancer free (CF) control group (age range 21–67). PON1 enzyme was prepared from the serum pool of BC patients using hydrophobic interaction chromatography on L-tyrosine-9-aminophenanthrene-coupled Sepharose 4Bgel. The PON1 enzyme activity towards paraoxon substrate was quantified spectrophotometrically. The basal activity of PON1 was statistically decreased in cancer cases compared to the control group. In addition, individuals were classified according to phenotyping of human PON1 Q and R types. In the cohort of BC patients, an increase in the frequency of the PON homozygote Q (AA) genotype was observed (31% in the BC group versus 14% in the CF controls). The frequency of the PON heterozygote QR (AB) genotype was 34.5% in the patients with BC and 37% in the CF group. The same trend was observed in PON homozygote R (BB) genotype frequency (BC cases 34.5% versus controls 49%). We determined that the kinetic parameters of the purified enzyme by Lineweaver–Burk method. We obtained K_m and V_max values of 0.227?mM and 62?U/mL?min for the BC enzyme, compared with 0.775?mM and 206?U/mL?min for the CF control enzyme. As a conclusion, it is clear from our results that while the PON1 AA allele frequency in BC cases is much higher, that of BB allele is much lower, in comparison with the control group. The most significant finding of this study is AA allele activity which is low in BC cases was found high. We concluded that decreased AA allele PON1 activity might have a relation with BC.     

Filling of eco-climatological niches in a polyploid complex – A case study in the plant genus Leucanthemum Mill. (Compositae,Anthemideae) from the Iberian Peninsula

In order to assess the effects of polyploidisation on distribution patterns of species in a polyploid complex, we compared potential and actual distribution ranges of representatives from the genus Leucanthemum Mill. (Compositae, Anthemideae), which occur on the Iberian Peninsula with 20 taxa spanning ploidy levels between 2n = 2x = 18 and 2n = 22x = 198. Actual distribution ranges of taxa were derived from geo-referenced herbarium specimens by defining buffer circles around the collection localities to account for geographical uncertainties and haphazardness of specimen collecting. Eco-climatologically potential ranges were modelled with the maximum entropy method (Maxent) based on the actual distributions. By comparison of sizes of realised and eco-climatologically potential areas for each taxon we computed taxon-specific overlap indices that express the filling of eco-climatological niches. The correlation between overlap indices and ploidy levels was tested by Spearman's rank correlation coefficient. We observed a significant negative correlation between ploidy level and sizes of realised distribution ranges and no significant correlation of ploidy level and sizes of potential ranges. Finally, we found that diploid and low-ploidy taxa are filling their potential eco-climatological distribution ranges more exhaustively than highly polyploid taxa. Assuming that ascending polyploidisation is the prevailing mode compared to descending polyploidisation, our results demonstrate that the distribution patterns of members of the Leucanthemum polyploid complex on the Iberian Peninsula are mainly influenced by the age of taxa and not by ecological advantages or disadvantages connected with polyploidy.     

Secondary structure of Aβ(1–16) complexes with zinc: A study in the gas phase using deuterium/hydrogen exchange and ultra-high-resolution mass spectrometry

Complexes of peptide fragment 1–16 of beta-amyloid with transition metals play an important role in the development of a broad class of neurodegenerative diseases, which determines the interest in investigating the structures of these complexes. In this work, we have applied the method of the deuterium/hydrogen exchange in combination with ultra-high-resolution mass spectrometry to study conformational changes in (1–16) beta-amyloid peptide induced by binding of zinc(II) atoms. The efficiency of the deuterium/hydrogen exchange depended on the number of zinc atoms bound to the peptide and on the temperature of the ionization source region. Deuterium/hydrogen exchange reactions have been performed directly in the ionization source. The number of exchanges decreased considerably with an increasing numbers of zinc atoms. The relationship has been described with a damped exponential curve, which indicated that the binding of zinc atoms altered the conformation of the peptide ion by making it less open, which limits the access to inner areas of the molecule.