Search

The fabrication of low-resistance and thermal stable ohmic contacts is important for realization of reliable SiC devices. For the n-type SiC, Ni-based metallization is most commonly used for Schottky and ohmic contacts. Many experimental studies have been performed in order to understand the mechanism of ohmic contact formation and different models were proposed to explain the Schottky to ohmic transition for Ni/SiC contacts. In the present review, we summarize the last key results on the matter and post open questions concerning the unclear issues of ohmic contacts to n-type SiC. Analysis of the literature data and our own experimental observations have led to the conclusion that the annealing at high temperature leads to the preferential orientation of silicide at the heterointerface (0001)SiC//(013)δ-Ni2Si. Moreover, we may conclude that only δ-Ni2Si grains play a key role in determining electrical transport properties at the contact/SiC interface. Finally, we show that the diffusion barriers with free diffusion path microstructure can improve thermal stability of metal-SiC ohmic contacts for high-temperature electronics.

RE-doped β-Ga 2 O 3 seems attractive for future high-power LEDs operating in high irradiation environments. In this work, we pay special attention to the issue of radiation-induced defect anisotropy in β-Ga 2 O 3 , which is crucial for device manufacturing. Using the RBS/c technique, we have carefully studied the structural changes caused by implantation and post-implantation annealing in two of the most commonly used crystallographic orientations of β-Ga 2 O 3 , namely the (-201) and (010). The analysis was supported by advanced computer simulations using the McChasy code. Our studies reveal a strong dependence of the structural damage induced by Yb-ion implantation on the crystal orientation, with a significantly higher level of extended defects observed in the (-201) direction than for the (010). In contrast, the concentration and behavior of simple defects seem similar for both oriented crystals, although their evolution suggests the co-existence of two different types of defects in the implanted zone with their different sensitivity to both, radiation and annealing. It has also been found that Yb ions mostly occupy the interstitial positions in β-Ga 2 O 3 crystals that remain unchanged after annealing. The location is independent of the crystal orientations. We believe that these studies noticeably extend the knowledge of the radiation-induced defect structure, because they dispel doubts about the differences in the damage level depending on crystal orientation, and are important for further practical applications., (© 2024. The Author(s).)

β-Ga 2 O 3 is an ultra-wide bandgap semiconductor (E g ~4.8 eV) of interest for many applications, including optoelectronics. Undoped Ga 2 O 3 emits light in the UV range that can be tuned to the visible region of the spectrum by rare earth dopants. In this work, we investigate the crystal lattice recovery of (2¯01)-oriented β-Ga 2 O 3 crystals implanted with Yb ions to the fluence of 1 ×10 14 at/cm 2 . Post-implantation annealing at a range of temperature and different atmospheres was used to investigate the β-Ga 2 O 3 crystal structure recovery and optical activation of Yb ions. Ion implantation is a renowned technique used for material doping, but in spite of its many advantages such as the controlled introduction of dopants in concentrations exceeding the solubility limits, it also causes damage to the crystal lattice, which strongly influences the optical response from the material. In this work, post-implantation defects in β-Ga 2 O 3 :Yb crystals, their transformation, and the recovery of the crystal lattice after thermal treatment have been investigated by channeling Rutherford backscattering spectrometry (RBS/c) supported by McChasy simulations, and the optical response was tested. It has been shown that post-implantation annealing at temperatures of 700-900 °C results in partial crystal lattice recovery, but it is accompanied by the out-diffusion of Yb ions toward the surface if the annealing temperature and time exceed 800 °C and 10 min, respectively. High-temperature implantation at 500-900 °C strongly limits post-implantation damage to the crystal lattice, but it does not cause the intense luminescence of Yb ions. This suggests that the recovery of the crystal lattice is not a sufficient condition for strong rare-earth photoluminescence at room temperature and that oxygen annealing is beneficial for intense infrared luminescence compared to other tested environments.

Although there has been no shortage of technological innovation in recent decades, a solution to sociodemographic disparities in the forensic setting has remained elusive. Artificial intelligence (AI) is a uniquely powerful emerging technology that is likely to either exacerbate or mitigate existing disparities and biases. This column argues that the implementation of AI in forensic settings is inevitable, and that practitioners and researchers should focus on developing AI systems that reduce bias and advance sociodemographic equity rather than attempt to impede the use of this novel technology., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Rare earth-doped zinc oxide (ZnO:RE) systems are attractive for future optoelectronic devices such as phosphors, displays, and LEDs with emission in the visible spectral range, working even in a radiation-intense environment. The technology of these systems is currently under development, opening up new fields of application due to the low-cost production. Ion implantation is a very promising technique to incorporate rare-earth dopants into ZnO. However, the ballistic nature of this process makes the use of annealing essential. The selection of implantation parameters, as well as post-implantation annealing, turns out to be non-trivial because they determine the luminous efficiency of the ZnO:RE system. This paper presents a comprehensive study of the optimal implantation and annealing conditions, ensuring the most efficient luminescence of RE 3+ ions in the ZnO matrix. Deep and shallow implantations, implantations performed at high and room temperature with various fluencies, as well as a range of post-RT implantation annealing processes are tested: rapid thermal annealing (minute duration) under different temperatures, times, and atmospheres (O 2 , N 2 , and Ar), flash lamp annealing (millisecond duration) and pulse plasma annealing (microsecond duration). It is shown that the highest luminescence efficiency of RE 3+ is obtained for the shallow implantation at RT with the optimal fluence of 1.0 × 10 15 RE ions/cm 2 followed by a 10 min annealing in oxygen at 800 °C, and the light emission from such a ZnO:RE system is so bright that can be observed with the naked eye.

The US Environmental Protection Agency's short-term freshwater effluent test methods include a fish (Pimephales promelas), a cladoceran (Ceriodaphnia dubia), and a green alga (Raphidocelis subcapitata). There is a recognized need for additional taxa to accompany the three standard species for effluent testing. An appropriate additional taxon is unionid mussels because mussels are widely distributed, live burrowed in sediment and filter particles from the water column for food, and exhibit high sensitivity to a variety of contaminants. Multiple studies were conducted to develop a relevant and robust short-term test method for mussels. We first evaluated the comparative sensitivity of two mussel species (Villosa constricta and Lampsilis siliquoidea) and two standard species (P. promelas and C. dubia) using two mock effluents prepared by mixing ammonia and five metals (cadmium, copper, nickel, lead, and zinc) or a field-collected effluent in 7-day exposures. Both mussel species were equally or more sensitive (more than two-fold) to effluents compared with the standard species. Next, we refined the mussel test method by first determining the best feeding rate of a commercial algal mixture for three age groups (1, 2, and 3 weeks old) of L. siliquoidea in a 7-day feeding experiment, and then used the derived optimal feeding rates to assess the sensitivity of the three ages of juveniles in a 7-day reference toxicant (sodium chloride [NaCl]) test. Juvenile mussels grew substantially (30%-52% length increase) when the 1- or 2-week-old mussels were fed 2 ml twice daily and the 3-week-old mussels were fed 3 ml twice daily. The 25% inhibition concentrations (IC25s) for NaCl were similar (314-520 mg Cl/L) among the three age groups, indicating that an age range of 1- to 3-week-old mussels can be used for a 7-day test. Finally, using the refined test method, we conducted an interlaboratory study among 13 laboratories to evaluate the performance of a 7-day NaCl test with L. siliquoidea. Eleven laboratories successfully completed the test, with more than 80% control survival and reliable growth data. The IC25s ranged from 296 to 1076 mg Cl/L, with a low (34%) coefficient of variation, indicating that the proposed method for L. siliquoidea has acceptable precision. Environ Toxicol Chem 2021;40:3392-3409. © 2021 SETAC., (© 2021 SETAC.)

The land cover reconstruction from monochromatic historical aerial images is a challenging task that has recently attracted an increasing interest from the scientific community with the proliferation of large-scale epidemiological studies involving retrospective analysis of spatial patterns. However, the efforts made by the computer vision community in remote-sensing applications are mostly focused on prospective approaches through the analysis of high-resolution multi-spectral data acquired by the advanced spatial programs. Hence, four contributions are proposed in this paper. They aim at providing a comparison basis for the future development of computer vision algorithms applied to the automation of the land cover reconstruction from monochromatic historical aerial images. First, a new multi-scale multi-date dataset composed of 4.9 million non-overlapping annotated patches of the France territory between 1970 and 1990 has been created with the help of geography experts. This dataset has been named HistAerial. Second, an extensive comparison study of the state-of-the-art texture features extraction and classification algorithms, including deep convolutional neural networks (DCNNs), has been performed. It is presented in the form of an evaluation. Third, a novel low-dimensional local texture filter named rotated-corner local binary pattern (R-CRLBP) is presented as a simplification of the binary gradient contours filter through the use of an orthogonal combination representation. Finally, a novel combination of low-dimensional texture descriptors, including the R-CRLBP filter, is introduced as a light combination of local binary patterns (LCoLBPs). The LCoLBP filter achieved state-of-the-art results on the HistAerial dataset while conserving a relatively low-dimensional feature vector space compared with the DCNN approaches (17 times shorter).

We report on AlxGa1-xN heterostructures resulting from the coherent growth of a positive then a negative gradient of the Al concentration on a [0001]-oriented GaN substrate. These polarization-doped p-n junction structures were characterized at the nanoscale by a combination of averaging as well as depth-resolved experimental techniques including: cross-sectional transmission electron microscopy, high-resolution X-ray diffraction, Rutherford backscattering spectrometry, and scanning probe microscopy. We observed that a small miscut in the substrate orientation along with the accumulated strain during growth led to a change in the mosaic structure of the AlxGa1-xN film, resulting in the formation of macrosteps on the surface. Moreover, we found a lateral modulation of charge carriers on the surface which were directly correlated with these steps. Finally, using nanoscale probes of the charge density in cross sections of the samples, we have directly measured, semiquantitatively, both n- and p-type polarization doping resulting from the gradient concentration of the AlxGa1-xN layers.

We propose a new coding technology for 3D video represented by multiple views and the respective depth maps. The proposed technology is demonstrated as an extension of the recently developed high efficiency video coding (HEVC). One base views are compressed into a standard bitstream (like in HEVC). The remaining views and the depth maps are compressed using new coding tools that mostly rely on view synthesis. In the decoder, those views and the depth maps are derived via synthesis in the 3D space from the decoded baseview and from data corresponding to small disoccluded regions. The shapes and locations of those disoccluded regions can be derived by the decoder without any side information transmitted. To achieve high compression efficiency, we propose several new tools such as depth-based motion prediction, joint high frequency layer coding, consistent depth representation, and nonlinear depth representation. The experiments show high compression efficiency of the proposed technology. The bitrate needed for transmission of two side views with depth maps is mostly less than 50% than that of the bitrate for a single-view video.

Recently published biochemical data suggest the significant role of selenium compounds as the adjuvants combined with L: -thyroxine therapy, which can reduce antithyroid peroxidase antibodies' (TPOAb) levels in patients with Hashimoto disease. The study was undertaken to document in a more detailed way the changes in parameters expressing the thyroid and ovarian function brought about by selenium supplementation (50-100 microg/day) in a woman undergoing autoimmune thyroiditis (AIT) therapy. This prospective observational case study lasted for 14 months plus additional 5 months as a follow-up period. Parameters reflecting selenium status, thyroid metabolism, and sex hormones secretion were determined at the onset and end of the study period, as well as in some of its middle points. During the supplementation trial, serum selenium (Se) increased by 45% and plasma glutathione peroxidase (GPX3) by 21%. TPOAb decreased by 76%. All other parameters also fluctuated during the supplementation period, but all results were always within normal physiological ranges. After withdrawal of the supplementation, the sharp fall of Se and GPX3 promptly occurred, and this phenomenon was accompanied with a marked increase in TPOAb. This report stresses the importance of selenium supplementation in AIT treatment. However, the efficiency and durability of the effect of Se supplementation on the TPOAb titer remain an open question. The clarification of mechanism(s) underlying Se interaction with autoimmune processes should throw new light on this issue.

The aim of the study was to identify those parameters characterising selenium status and sex hormones secretion, which are responsible for the changes in indicators of thyroid function, and to reveal the correlation structure of parameters expressing selenium status, sex hormones secretion and thyroid function, if any exist. The general approach in this work was essentially the same as in our previous report (cf. [Zagrodzki P, Ratajczak R, Wietecha-Posłuszny R. The interaction between selenium status, sex hormones, and thyroid metabolism in adolescent girls during the luteal phase of their menstrual cycle. Biol Trace Elem Res 2007; 120: 51-60]), but the study group and parameters' spectrum were different. Thirty-six women aged 23.5+/-0.6 years were investigated. The parameters of interest were: plasma selenium concentration (Se) and plasma glutathione peroxidase activity (GPX3) (indicators of selenium status); serum estradiol (E2), progesterone (P4), follicle-stimulating hormone (FSH), luteotropic hormone (LH), and age of menarche (indicators of sex hormones secretion); thyroid-stimulating hormone (TSH), free thyroxine (fT4), free triiodothyronine (fT3), anti-thyroid peroxidase antibodies in serum (anti-TPO), and thyroid volume (indicators of thyroid function). Our findings show that Se, GPX3, P4, LH, FSH, TSH and fT4 were interrelated. The pairs of parameters: GPX3 and P4; LH and FSH exhibited the strongest associations. The thyroid function parameters were mainly influenced by GPX3 and P4. Partial least-squares method corroborated our hypothesis about the existence of complex interactions among selenium status, sex hormones secretion, and thyroid function in healthy young women in the luteal phase.

The objective of the present work was to study all physiological relationships among selenium status (SeS), sex hormones secretion (SH), and thyroid metabolism (ThM) in healthy adolescent girls, at one time. Forty-four girls aged 13.4-16.6 years (mean age, 14.5 +/- 0.5 years) entered the statistical model. Parameters reflecting SeS: plasma selenium concentration (Se) and plasma glutathione peroxidase activity (GPX3); SH: serum estradiol (E2) and progesterone (P4); age of menarche (AoM); and ThM: thyroid stimulating hormone (TSH), free thyroxine (fT4), free triiodothyronine (fT3), antithyroid peroxidase antibodies (anti-TPO) in serum, and thyroid volume (ThV), were determined, and the interactions between them were evaluated by means of the partial least squares method (PLS). PLS method was, for the first time, successfully applied to the problem of selenium and hormone interactions and revealed that selenium status and female reproductive system are interrelated and affect thyroid physiology in adolescent girls in the luteal phase. The strongest associations were revealed for the pairs of parameters, Se and fT4/fT3, Se and P4, the modest ones for the pairs, Se and ThV, P4 and fT4/fT3, Se and AoM, and P4 and AoM. There was no correlation between E2, GPX3, and TSH, and any other considered parameter. Se and P4 had the greatest influence on ThM parameters.

Modification of materials is a wide area in materials science, especially surface modification. To investigate the results of the modification process, treated and nontreated samples were compared. Intense plasma pulses of argon or nitrogen were used to irradiate the carbon steels. In all samples, the near-surface layer was melted. Results of scanning electron microscopy investigations of the surface morphology and cross-sections, as well as the results of tribological tests, are presented. The obtained results allowed us to draw conclusions about changes in material properties and to propose subsequent studies using other investigation techniques.

The halophyte Suaeda salsa L., exposed to different NaCl concentrations (100 and 400 mmol/L) and polyethylene glycol (isoosomotic to 100 mmol/L NaCl) containing nutrient solutions under normal or K+-deficient conditions for 7 days, was used to study effects of NaCl salinity and osmotic stress on chlorophyll content, chlorophyll fluorescence characteristics, malonedialdehyde (MDA) content, and superoxide dismutase (SOD) isoform activities. Photosynthetic capacity was not decreased by NaCl treatment, indicating that S. salsa possesses an effective antioxidative response system for avoiding oxidative damage. Seven SOD activity bands were detected in S. salsa leaf extracts, including an Mn-SOD and several isoforms of Fe-SOD and CuZn-SOD. It turned out that NaCl salinity and osmotic stress lead to a differential regulation of distinct SOD isoenzymes. This differential regulation is suggested to play a major role in stress tolerance of S. salsa.

Citrus juice cells accumulate high levels of citric acid in their vacuoles when compared to other organic ions including malate. Uptake of citrate into tonoplast vesicles from Citrus juice cells was investigated in the presence of malate, and after incubation with antibodies raised against the vacuolar malate-specific channel of Kalanchoë diagremontiana leaves. Antibodies against the vacuolar malate channel immunoreacted with a protein of similar size in tonoplast extracts from three Citrus varieties differing in citric acid content. Malate channel antibodies inhibited both delta MicroH(+)-dependent and delta MicroH(+)-independent ATP-dependent citrate transport, indicating common domains in both transport systems and to the malate-specific channel of Kalanchoë diagremontiana leaves. Malate strongly inhibited electrogenic citrate transport, whereas ATP-dependent citrate uptake was less affected. Kinetic analysis of citrate transport in the presence of malate confirmed the existence of two citrate transport mechanisms and indicated that both citrate and malate share a common transport channel across the tonoplast of Citrus juice cells.

Thyroid hormones are very important for the child mental and physical development. In southeast Poland screening for congenital hypothyroidism detection was established in 1985; this region was defined as an iodine deficient area. The results provided the first signal pointing to insufficient iodine prophylaxis in the region. In the years 1985-2000, a total of 634,179 newborns were tested. TSH levels in blood spots on filter paper were determined using three consecutive methods: RIA, fluoroimmunometry (FIA) and luminoimmunemetry (LIA). In the first five years of the program, the incidence of permanent hypothyroidism was 1:4,000, while the incidence of iodine deficiency disorders (IDD) was 1:272. These findings led to the recognition of the fact that iodine prophylaxis had been discontinued in 1981 and reintroduced in 1992 and from that time on the incidence of IDD in neonates has been evidently lower. In the period between 1991 and 2000, we observed a 0.14% incidence rate of IDD, while until 1991 it had been 0.5%. In the years 1998-2000, the respective value was 1:5,420. The incidence of transient hypothyroidism also dropped from 1:3,920 to 1:48,474. Therefore we conclude that, based on screening studies carried out since 1985, the analysis of TSH levels may be regarded as a tool for iodine deficiency monitoring in neonates. Between 1985-2000, a drop in the incidence of IDD in newborns is clearly seen. Moreover, investigations carried out in particular years show that even low-grade iodine supplementation leads to TSH level decrease in newborns.

The model plant tobacco (Nicotiana tabacum L.) was chosen for a survey of the subunit composition of the V-ATPase at the protein level. V-ATPase was purified from tobacco leaf cell tonoplasts by solubilization with the nonionic detergent Triton X-100 and immunoprecipitation. In the purified fraction 12 proteins were present. By matrix-assisted laser-desorption ionization mass spectrometry (MALDI-MS) and amino acid sequencing 11 of these polypeptides could be identified as subunits A, B, C, D, F, G, c, d and three different isoforms of subunit E. The polypeptide which could not be identified by MALDI analysis might represent subunit H. The data presented here, for the first time, enable an unequivocal identification of V-ATPase subunits after gel electrophoresis and open the possibility to assign changes in polypeptide composition to variations in respective V-ATPase subunits occurring as a response to environmental conditions or during plant development.

V-ATPases pump protons into the interior of various subcellular compartments at the expense of ATP. Previous studies have shown that these pumps comprise a membrane-integrated, proton-translocating (V(0)), and a soluble catalytic (V(1)) subcomplex connected to one another by a thin stalk region. We present two three-dimensional maps derived from electron microscopic images of the complete V-ATPase complex from the plant Kalanchoë daigremontiana at a resolution of 2.2 nm. In the presence of a non-hydrolyzable ATP analogue, the details of the stalk region between V(0) and V(1) were revealed for the first time in their three-dimensional organization. A central stalk was surrounded by three peripheral stalks of different sizes and shapes. In the absence of the ATP analogue, the tilt of V(0) changed with respect to V(1), and the stalk region was less clearly defined, perhaps due to increased flexibility and partial detachment of some of the peripheral stalks. These structural changes corresponded to decreased stability of the complex and might be the initial step in a controlled disassembly.

The metabolic switch from C3-photosynthesis to crassulacean acid metabolism (CAM),and the antioxidative response of Mesembryanthemum crystallinum L. plants cultured under severe salt stress and high light intensities, and a combination of both stress conditions, were studied. High light conditions led to a more rapid CAM induction than salinity. The induction time was still shortened when both stress factors were combined. A main pattern observed in CAM plants was a decrease in mitochondrial Mn-superoxide dismutase (SOD) activity during the day. The activities of the chloroplastic Fe-SOD and cytosolic CuZn-SOD were increased due to salt treatment after a lag phase, while catalase activity was decreased. Combination of salt and light stress did not lead to a higher SOD activity as found after application of one stress factor alone, indicating that there is a threshold level of the oxidative stress response. The fact that salt-stressed plants grown under high light conditions showed permanent photoinhibition and lost the ability for nocturnal malate storage after 9 d of treatment indicate serious malfunction of metabolism, leading to accelerated senescence. Comparison of CuZn-SOD activity with CuZn-SOD protein amount, which was determined immunologically, indicates that the activity of the enzyme is at least partially post-translationally regulated.

The Chenopodiaceae Suaeda salsa L. was grown under different salt concentrations and under osmotic stress. The fresh weight was markedly stimulated by 0.1 M NaCl, 0.4 M NaCl and 0.1 M KCl and reduced by osmotic stress (PEG iso-osmotic to 0.1 M NaCl). Treatment with 0.4 M KCl severely damaged the plants. Membrane vesicle fractions containing tonoplast vesicles were isolated by sucrose gradient from leaves of the S. salsa plants and modulations of V-ATPase and V-PPase depending on the growth conditions were determined. Western blot analysis revealed that V-ATPase of S. salsa consists of at least nine subunits (apparent molecular masses 66, 55, 52, 48, 36, 35, 29, 18, and 16 kDa). This polypeptide pattern did not depend on culture conditions. V-PPase is composed of a single polypeptide (69 kDa). An additional polypeptide (54 kDa) was detected in the fractions of NaCl-, KCl- and PEG-treated plants. It turned out that the main strategy of salt-tolerance of S. salsa seems to be an up-regulation of V-ATPase activity, which is required to energize the tonoplast for ion uptake into the vacuole, while V-PPase plays only a minor role. The increase in V-ATPase activity is not obtained by structural changes of the enzyme, but by an increase in V-ATPase protein amount.

Plasma-membrane-located primary pumps were investigated in the sieve element (SE)-companion cell complex in the transport phloem of 2-week-old stems of Ricinus communis L. and, for comparison, in stems of Cucurbita pepo L. and in the secondary phloem of Agrobacterium tumefaciens-induced crown galls as a typical sink tissue. The plasma-membrane (PM) H+-ATPase and the tonoplast-type pyrophosphatase (PPase) were immunolocalized by epifluorescence and confocal laser scanning microscopy (CLSM) upon single or double labeling with specific monoclonal and polyclonal antibodies. Quantitative fluorescence evaluation by CLSM revealed both pumps in one membrane, the sieve-element PM. Different PM H+-ATPase antibody clones, raised against the PM H+-ATPase of Zea mays coleoptiles, induced in mouse and produced in mouse hybridoma cells, discriminated between different phloem cell types. Clones 30D5C4 and 44B8A1 labeled sieve elements and clone 46E5B11D5 labeled companion cells, indicating the existence of different phloem PM H+-ATPase isoforms. The results are discussed in terms of energization of SE transporters for retrieval of leaking sucrose, K+ and amino acids, as one of the unknown roles of ATP found in SEs. The function of the PPase could be related to phloem sucrose metabolism in support of ATP-requiring processes.

Anion uptake by isolated tonoplast vesicles was recorded indirectly via increased H(+)-transport by H(+)-pumping of the V-ATPase due to dissipation of the electrical component of the electrochemical proton gradient, Deltamu(H+), across the membrane. ATP hydrolysis by the V-ATPase was measured simultaneously after the Palmgren test. Normalizing for ATP-hydrolysis and effects of chloride, which was added to the assays as a stimulating effector of the V-ATPase, a parameter, J(mal)(rel), of apparent ATP-dependent malate-stimulated H(+)-transport was worked out as an indirect measure of malate transport capacity. This allowed comparison of various species and physiological conditions. J(mal)(rel) was high in the obligate crassulacean acid metabolism (CAM) species Kalanchoë daigremontiana Hamet et Perrier, it increased substantially after CAM induction in ice plant (Mesembryanthemum crystallinum), and it was positively correlated with NO(3)(-) nutrition in tobacco (Nicotiana tabacum). For tobacco this was confirmed by measurements of malate transport energized via the V-PPase. In ice plant a new polypeptide of 32-kD apparent molecular mass appeared, and a 33-kD polypeptide showed higher levels after CAM induction under conditions of higher J(mal)(rel). It is concluded that tonoplast malate transport capacity plays an important role in physiological regulation in CAM and NO(3)(-) nutrition and that a putative malate transporter must be within the 32- to 33-kD polypeptide fraction of tonoplast proteins.

Soluble proteins were isolated from leaves of the common ice plant Mesembryanthemum crystallinum L. in the CAM state of photosynthesis and tested for protease activity using amino acid-beta-naphthylamide (NA)-derivatives in a search for proteolytic activity responsible for cleavage of the V-ATPase subunit B. This cleavage is suggested to occur at the peptide bond between Met192 and Glu193. At neutral pH Met-NA was one of seven derivatives which were cleaved by proteases present in this fraction. Enzymes exhibiting proteolytic activity were separated from other soluble proteins by Superose 12-size exclusion FPLC. Incubation of partially purified protease with tonoplast-enriched membrane vesicle fractions isolated from M. crystallinum in the C3-state of photosynthesis led to a decrease in subunit B (55 kDa) protein amount and to the formation of the polypeptide Di (32 kDa), which has been previously suggested to represent a fragment of subunit B. Cleavage of subunit B and the appearance of Di also occurred during incubation of tonoplast vesicles in the presence of reactive oxygen species. In addition to Di, the polypeptide Ei (28 kDa) appeared after incubation with protease and/or reactive oxygen species. Taken into account that Di and Ei cross-reacted with an affinity purified antiserum directed against subunit B, Di as well as Ei might represent fragments of subunit B. These results open new perspectives with respect to the regulation of V-ATPase modification and turnover.

The plant V-ATPase is a primary-active proton pump present at various components of the endomembrane system. It is assembled by different protein subunits which are located in two major domains, the membrane-integral V(o)-domain and the membrane peripheral V(1)-domain. At the plant vacuole the V-ATPase is responsible for energization of transport of ions and metabolites, and thus the V-ATPase is important as a 'house-keeping' and as a stress response enzyme. It has been shown that transcript and protein amount of the V-ATPase are regulated depending on metabolic conditions indicating that the expression of V-ATPase subunit is highly regulated. Moreover, there is increasing evidence that modulation of the holoenzyme structure might influence V-ATPase activity.

Possible interactions between selenium and iodine metabolism were investigated in 7- to 16-year-old children with goiter (n = 136) living in southeastern Poland in iodine-deficient areas influenced by a sulfur industry. The Se-iodine interactions in these children were compared to the interactions in children from outside of that region (n = 38). Blood selenium (BSe) concentration and plasma glutathione peroxidase activity were much lower in the study group (64.1 +/- 15.7 microg/L; 111.0 +/- 27.6 U/L) than in the control group (85.3 +/- 19.6 microg/L; 182.4 +/- 35.6 U/L). Almost all of the data [plasma thyroid-stimulating hormone (TSH) concentration, plasma free thyroxine (fT(4)) concentration] fell within the reference limits. There was no statistically significant difference between the control and the study groups with respect to fT(4) and TSH. However, statistically significant differences of fT(4) and TSH in the study group were revealed between females belonging to the lower (n = 21; fT(4), 16.1 +/- 3.3 pmol/L; TSH, 1.83 +/- 1.05 mU/L) and upper Se quartiles (n = 24; fT(4), 14.5 +/- 2.2 pmol/L; TSH, 1. 26 +/- 0.90 mU/L), p < 0.05. Neither group differed in iodine in urine concentration, age, and body mass index. The difference in fT(4) concentrations can be attributed to an Se deficiency. The relationship exists only for females, which suggests a sex-linked hormonal response to concomitant Se and iodine deficiencies.

Treatment with 0.4 mol dm(-3) NaCl caused a C3-CAM shift in Mesembryanthemum crystallinum L. leaves. In parallel to the CAM induction the activity of CAT was significantly decreased. In C3 and in CAM plants CAT activity showed daily fluctuations, with the maximum at the end of the light period. The oscillations of CAT were more pronounced in CAM than in C3 plants. In M. crystallinum CAT activity seems to respond more to CAM induction than to salinity.

The success of a screening program depends in great measure on effective organization. The purpose of the screening program is the commencement of treatment of selected patients within two weeks of life. In total in our laboratory we have tested 461,479 newborns during 1985-1996. The introduction of bar codes in 1995 (recommended by the Institute of Mother and Child in Warsaw) improved our work in the laboratory and departments of obstetrics and reduced the possibility of errors. The screening tests for congenital hypothyroidism appear to serve the monitoring of iodine deficiency disorders. The good organization of the screening program is indispensable and needs permanent control.

Using a polyclonal antiserum specific for the tonoplastic H(+)-pyrophosphatase (tPPase), significant amounts of antigenic polypeptides of the correct molecular mass were detected in Western blots of plasma membrane isolated from cauliflower (Brassica oleracea L.) inflorescence by phase-partitioning and subsequent sucrose density centrifugation. Potassium iodide-stripped plasma membranes continued to give a strong positive signal, indicating that the PPase antigen detected was not a result of contamination through soluble PPase released during homogenisation. The same preparation contained negligible vacuolar (v)H(+)-ATPase activity and the A subunit of the vATPase could not be detected by immunoblotting. Plasma membrane fractions exhibited a proton-pumping activity with ATP as substrate, but such an activity was not measurable with pyrophosphate, although the hydrolysis of this substrate was recorded. By contrast, pyrophosphate supported proton pumping in tonoplast-containing fractions. Immunogold electron microscopy confirmed the presence of PPase at the plasma membrane as well as at the tonoplast, trans Golgi network, and multivesicular bodies. The density of immunogold label was higher at the plasma membrane than at the tonoplast, except for membrane fragments occurring in the lumen of the vacuoles which stained very conspicuously.

The present study deals with the phenotypic adaptation of tonoplast fluidity in the CAM plant Kalanchoë daigremontiana to changes in growth temperature. Tonoplast fluidity was characterized by measuring fluorescence depolarization in membranes labeled with fluorescent fatty acid analogues and by following formation of eximeres in membranes labeled by eximere-forming fluorophores. With both techniques it was found that exposure of the plants to higher growth temperature compared with the control decreased the fluidity of the tonoplast while exposure to lower growth temperature caused the opposite. Three hours of high temperature treatment (raised from 25 degreesC to 35 degreesC; "heat shock") were sufficient to decrease the tonoplast fluidity to roughly the same extent as growth under high temperature for 30 days. The phenotypic response of tonoplast fluidity to changes in growth temperature was found only in the complete membrane, not however in the lipid matrix deprived of the membrane proteins. Heat treatments of the plants decreased the lipid/protein ratio while exposure to low temperature (for 30 days) increased it. Heat treatments led to a decrease in the percentage of linolenic acid (C18:3) and linoleic acid (C18:2), heat shock and low temperature treatments induced an increase in the percentage of linoleic acid (C18:3), with concomitant decrease in the percentage of linoleic acid (C18:2). However, in the case of heat shock, increase in linolenic acid concerned mainly monogalactosyldiacylglycerol, while with low temperature treatment linoleic acid increased in phosphatidylcholine. Both treatment of the plants with high and low temperature led to a slight decrease in the contribution of phosphatidylcholine and phosphoethanolamine to the total phospholipid content of the tonoplast. High-temperature treatment of the plants not only decreased the phospholipid/protein ratio in the tonoplast, but also led to the occurrence of a 35 kDa polypeptide in the tonoplast which cross-reacted with an antiserum against the tonoplast H+-ATPase holoenzyme. The important role of membrane proteins in bringing about the phenotypic rigidization of the tonoplast was mimicked by reconstitution experiments showing that incorporation of the proteins isolated from the tonoplast into phosphatidylcholine vesicles decreased the fluidity of this membrane system. As to be expected from the analyses in the natural membrane, the degree of this effect depended on the phospholipid/protein ratio.

Two antisera (ATP88 and ATP95) raised against the V-ATPase holoenzyme of Kalanchoë daigremontiana were tested for their cross-reactivity with subunits of V-ATPases from other plant species. V-ATPases from Kalanchoë blossfeldiana, Mesembryanthemum crystallinum, Nicotiana tabacum, Lycopersicon esculentum, Citrus limon, Lemna gibba, Hordeum vulgare and Zea mays were immunoprecipitated with an antiserum against the catalytic V-ATPase subunit A of M. crystallinum. As shown by silver staining and Western blot analysis with ATP88, subunits A, B, C, D and c were present in all immunoprecipitated V-ATPases. In contrast, ATP95 recognized the whole set of subunits only in K. blossfeldiana, M. crystallinum, H. vulgare and Z. mays. This differential cross reactivity of ATP95 indicates the presence of structural differences of certain V-ATPase subunits. Based on the Bafilomycin A1-sensitive ATPase activity of tonoplast enriched vesicles, and on the amount of V-ATPase solubilized and immunoprecipitated, the specific ATP-hydrolysis activity of the V-ATPases under test was determined. The structural differences correlate with the ability of V-ATPases from different species to hydrolyze ATP at one given assay condition for ATP-hydrolysis measurements. Interestingly V-ATPases showing cross-reactivity of subunits A, B, C, D and c with ATP95 showed higher rates of specific ATP hydrolysis compared to V-ATPases containing subunits which were not labeled by ATP95. Thus, V-ATPases with high turnover rates in our assay conditions may show common structural characteristics which separate them from ATPases with low turnover rates.

Mature motor cells of Mimosa pudica that exhibit large and rapid turgor variations in response to external stimuli are characterized by two distinct types of vacuoles, one containing large amounts of tannins (tannin vacuole) and one without tannins (colloidal or aqueous vacuole). In these highly specialized cells we measured the abundance of two tonoplast proteins, a putative water-channel protein (aquaporin belonging to the [gamma]-TIPs [tonoplast intrinsic proteins]) and the catalytic A-subunit of H+-ATPase, using either high-pressure freezing or chemical fixation and immunolocalization. [gamma]-TIP aquaporin was detected almost exclusively in the tonoplast of the colloidal vacuole, and the H+-ATPase was also mainly localized in the membrane of the same vacuole. Cortex cells of young pulvini cannot change shape rapidly. Development of the pulvinus into a motor organ was accompanied by a more than 3-fold increase per length unit of membrane in the abundance of both aquaporin and H+-ATPase cross-reacting protein. These results indicate that facilitated water fluxes across the vacuolar membrane and energization of the vacuole play a central role in these motor cells.

In the facultative halophyte Mesembryanthemum crystallinum, the salt- or age-induced transition to crassulacean acid metabolism (CAM) leads to the occurrence of a tonoplast-bound 32 kDa polypeptide (Di). The alignment of its N-terminal protein sequence with protein sequences of recently cloned higher plant V-ATPase B-subunits indicates that Di may be derived from subunit B by proteolytic removal of a protein fragment of about 20 kDa from its N-terminus. Furthermore, an antiserum directed against Di cross-reacts with subunit B from Nicotiana tabacum. It inhibits both proton pumping and ATP hydrolysis of the holoenzyme in M. crystallinum. As Di remains firmly attached to the holoenzyme the proteolytic processing may have functional implications.

In Mesembryanthemum crystallinum, the salt stress-induced metabolic switch from C3 photosynthesis to Crassulacean acid metabolism is accompanied by major changes in gene expression. However, early effects of salt exposure (i.e. prior to Crassulacean acid metabolism induction) on genes coding for vacuolar transport functions have not yet been studied. Therefore, the expression of vacuolar H(+)-ATPase genes was analyzed in different organs of 4-week-old plants stressed with 400 mM NaCl for 3, 8, or 24 h. Partial cDNAs for the subunits A, B, and c were cloned and used as homologous probes for northern blot analysis. In control plants, the mRNA levels for the different subunits showed organ-specific differences. In fully expanded leaves, subunit c mRNA was very low but increased transiently during the light period. Plant organs also differed in their salt-stress response. In roots and young leaves, mRNA levels for all three subunits increased about 2-fold compared to control plants, whereas in fully expanded leaves only subunit c mRNA responded to salt. The results indicate that the expression of vacuolar H(+)-ATPase genes does not always involve a fixed stoichiometry of mRNAs for the different subunits and that the mRNA level for subunit c is particularly sensitive to developmental and environmental changes.

The region of Southern Poland is known as an iodine-deficient area. Iodine prophylaxis was discontinued in 1980/81. This fact was discovered as a result of the introduction of newborn mass screening for congenital hypothyroidism in 1985. In view of the large number of newly detected cases of Iodine Deficiency Disorders (IDD), we compared the neonatal populations of the Cracow and Nowy Sacz provinces. The prevalence of IDD was revealed as eight times higher in Nowy Sacz than in Cracow. The iodine urine excretion rates in those two provinces were similar. The paper discusses the cause of the discrepancy in IDD incidence. The final results show that the iodine deficiency detected in the Cracow and Nowy Sacz provinces is of moderate degree. Salt iodination in the region of southern Poland needs a new approach even before the year 2000, as planned in the WHO program.

The study included 4103 school children attending randomly selected schools localized in towns and villages of 3 geographically distinct regions: mountainous, (Carpathian endemia), upland, lowland. The children were divided into four age-related groups. The incidence of goiter as detected by palpation was 38.1% in all the children studied, the value being similar in all three regions (38.1, 37.3 and 38.6%, respectively). Goiter incidence differs slightly among urban and rural children, amounting to 40.5 and 34.5%, respectively, in the two groups. In about 60% of children urinary iodine excretion was lower than 50 micrograms/l, while in about 30% it was normal. In about 10% of children studied urinary iodine excretion was lower than 20 micrograms/l. The results obtained indicate the occurrence among children of the studied region of goiter endemy of moderate degree. It was found that the percentage of children excreting more than 50 micrograms/l of urinary iodine was higher among those consuming iodized salt as compared to those consuming non-iodized salt. The first effects of resumption of common salt fortification with iodide in southern Poland can already be noted. There is, however, a need for optimization and steady control over this prophylactic action.

The study on the effects of Czarnobyl accident on thyroid gland function in children (MZ XVII program) revealed a high incidence of goiter in the population of children in district Kraków and Nowy Sacz. Other study on Tarnobrzeg population showed that the frequency of goiter in school children was 67%. The goiter prevalence, urine iodine excretion and iodine food consumption in the same populations were compared. The detailed investigation of iodine intake in children by feeding questionnaire shows a low consumption of iodine. The urine iodine excretion in the population of Kraków-district was low, but higher than in Nowy Sacz-district and nearly normal in Tarnobrzeg. High frequency of goiter in children and of IDD in newborns screened for CH, as well as low urine iodine excretion, together with low iodine intake with food are the markers of iodine deficiency in Kraków and Nowy Sacz districts. The improvement of feeding as well as iodine salt supplementation is necessary. The situation in Tarnobrzeg district looks differently and needs additional study.

Screening for congenital hypothyroidism has been carried out in 69,286 newborn children born in the region and city of Krakow in the years 1985-1988. Fourteen cases of congenital thyroid insufficiency were detected indicating the incidence of 1:4816 in this region. In 25 newborns (incidence of 1:1979) a transient hyperthyreotropinemia was diagnosed. It was concluded that the main reason of the appearance of an elevated TSH level in newborns of this region seems to be the deficiency of iodine in the diet and/or environment.

The lateral distribution of plastocyanin in the thylakoid lumen of spinach and pea chloroplasts was studied by combining immunocytochemical localization and kinetic measurements of P700+ reduction at high time resolution. In dark-adapted chloroplasts, the concentration of plastocyanin in the photosystem I containing stroma membranes exceeds that in photosystem II containing grana membranes by a factor of about two. Under these conditions, the reduction of P700+ with a halftime of 12 microseconds after a laser flash of saturating intensity indicates that to greater than 95% of total photosystem I a plastocyanin molecule is bound. An analysis of the labeling densities, the length of the different lumenal regions, and the total amounts of plastocyanin and P700 shows that most of the remaining presumable mobile plastocyanin is found in the granal lumen. This distribution of plastocyanin is consistent with a more negative surface charge density in the stromal than in the granal lumen. During illumination the concentration of plastocyanin in grana increases at the expense of that in stroma lamellae, indicating a light-driven diffusion from stroma to grana regions. Our observations provide evidence that a high concentration of plastocyanin in grana in the light favors the lateral electron transport from cytochrome b6/f complexes in appressed grana across the long distance to photosystem I in nonappressed stroma membranes.