Ultrastructural Modifications Research Articles

The Impact Assessment of CuO Nanoparticles on the Composition and Ultrastructure of Triticum aestivum L.

In the present study, the effects of copper oxide nanoparticles (CuO NPs) on bioactive compounds, the ultrastructural modifications which can occur, and elemental content of wheat were investigated. Changes in the wheat plants grown in presence or absence of CuO NPs were estimated. The application of CuO NPs decreased the amounts of chlorophylls and carotenoids and increased the amounts of polyphenols and antioxidant capacity. Ultrastructural analysis showed that the plants treated with CuO NPs were negatively affected. Soil amending completely inhibited the accumulation of seventeen elements, while K, Br, Al, and Zn were accumulated and Cl, Na, Ba, and Sr content decreased in wheat samples, regardless of the type of NPs applied. The application of chemically obtained NPs induced the most significant changes, completely blocking the assimilation of Fe, Mo, As, Sb, and Sm, and favoring much higher accumulation of Br than biogenic NPs. The decrease in chlorophylls and carotenoids is correlated with increase in antioxidant capacity, and occurs with increase of Mo, Al, Mg, K, Zn, and Ca content. The behavior of total polyphenols is correlated with Br content, and antagonist to Al behavior. From the point of view of bioactive compounds, the most affected plants were those that grew in the presence of CuO-NP-cel, while from the point of view of elementary analysis, the most affected plants were those grown in the presence of CuO-NP. By corroborating the obtained results, it was found that the CuO NPs have a negative effect on wheat plants.

Rapid Detection of Imipenem Resistance in Gram-Negative Bacteria Using Tabletop Scanning Electron Microscopy: A Preliminary Evaluation.

Background: Enabling faster Antimicrobial Susceptibility Testing (AST) is critical, especially to detect antibiotic resistance, to provide rapid and appropriate therapy and to improve clinical outcomes. Although several standard and automated culture-based methods are available and widely used, these techniques take between 18 and 24 h to provide robust results. Faster techniques are needed to reduce the delay between test and results.Methods: Here we present a high throughput AST method using a new generation of tabletop scanning electron microscope, to evaluate bacterial ultra-structural modifications associated with susceptibilities to imipenem as a proof of concept. A total of 71 reference and clinical strains of Gram-negative bacteria were used to evaluate susceptibility toward imipenem after 30, 60, and 90 min of incubation. The length, width and electron density of bacteria were measured and compared between imipenem susceptible and resistant strains.Results: We correlated the presence of these morphological changes to the bacterial susceptibility and their absence to the bacterial resistance (e.g., Pseudomonas aeruginosa length without [2.24 ± 0.61 μm] and with [2.50 ± 0.68 μm] imipenem after 30 min [p = 3.032E-15]; Escherichia coli width without [0.92 ± 0.07 μm] and with [1.28 ± 0.19 μm] imipenem after 60 min [p = 1.242E-103]). We validated our method by a blind test on a series of 58 clinical isolates where all strains were correctly classified as susceptible or resistant toward imipenem.Conclusion: This method could be a potential tool for rapidly identifying carbapenem-resistance in Enterobacterales in clinical microbiology laboratories in <2 h, allowing the empirical treatment of patients to be rapidly adjusted.

The cytostome-cytopharynx complex of intracellular and extracellular amastigotes of Trypanosoma cruzi exhibit structural and functional differences.

Endocytosis in Trypanosoma cruzi is mainly performed through a specialised membrane domain called cytostome-cytopharynx complex. Its ultrastructure and dynamics in endocytosis are well characterized in epimastigotes, being absent in trypomastigotes, that lack endocytic activity. Intracellular amastigotes also possess a cytostome-cytopharynx but participation in endocytosis of these forms is not clear. Extracellular amastigotes can be obtained from the supernatant of infected cells or in vitro amastigogenesis. These amastigotes share biochemical and morphological features with intracellular amastigotes but retain trypomastigote's ability to establish infection. We analysed and compared the ultrastructure of the cytostome-cytopharynx complex of intracellular amastigotes and extracellular amastigotes using high-resolution tridimensional electron microscopy techniques. We compared the endocytic ability of intracellular amastigotes, obtained through host cell lysis, with that of extracellular amastigotes. Intracellular amastigotes showed a cytostome-cytopharynx complex similar to epimastigotes'. However, after isolation, the complex undergoes ultrastructural modifications that progressively took to an impairment of endocytosis. Extracellular amastigotes do not possess a cytostome-cytopharynx complex nor the ability to endocytose. Those observations highlight morpho functional differences between intra and extracellular amastigotes regarding an important structure related to cell metabolism. TAKE AWAYS: T. cruzi intracellular amastigotes endocytose through the cytostome-cytopharynx complex. The cytostome-cytopharynx complex of intracellular amastigotes is ultrastructurally similar to the epimastigote. Intracellular amastigotes, once outside the host cell, disassembles the cytostome-cytopharynx membrane domain. Extracellular amastigotes do not possess a cytostome-cytopharynx either the ability to endocytose.

Antifungal and Anti-Inflammatory Potential of Bupleurum rigidum subsp. paniculatum (Brot.) H.Wolff Essential Oil.

Fungal infections remain a major health concern with aromatic plants and their metabolites standing out as promising antifungal agents. The present study aims to assess, for the first time, the antifungal and anti-inflammatory potential of Bupleurum subsp. paniculatum (Brot.) H.Wolff essential oil from Portugal. The oil obtained by hydrodistillation and characterized by GC-MS, showed high amounts of monoterpene hydrocarbons, namely α-pinene (29.0–36.0%), β–pinene (26.1–30.7%) and limonene (10.5–13.5%). The antifungal potential was assessed, according to CLSI guidelines, against several clinical and collection strains. The essential oil showed a broad fungicidal effect being more potent against Cryptococcus neoformans and dermatophytes. Moreover, a significant germ tube inhibition was observed in Candida albicans as well as a disruption of mature biofilms, thus pointing out an effect of the oil against relevant virulent factors. Furthermore, fungal ultrastructural modifications were detected through transmission electron microscopy, highlighting the nefarious effect of the oil. Of relevance, the oil also evidenced anti-inflammatory activity through nitric oxide inhibition in macrophages activated with lipopolysaccharide. In addition, the essential oil’s bioactive concentrations did not present toxicity towards macrophages. Overall, the present study confirmed the bioactive potential of B. rigidum subsp. paniculatum essential oil, thus paving the way for the development of effective drugs presenting concomitantly antifungal and anti-inflammatory properties.

Biotransformation of Non-steroidal Anti-inflammatory Drugs Induces Ultrastructural Modifications in Green Leafy Vegetables

More than we would like to admit, pharmaceutical products represent a high risk for the environment since, in the past decades, their concentration in natural, drinking, and irrigation waters reached concerning concentrations. The threat also resides in the fact that they tend to accumulate in edible plants and crops, and in the end, they could affect other organisms in the food chain, including humans. The aim of the work was to address some of these probable threats of pharmaceutical drugs. Therefore, the effects of three essential non-steroidal anti-inflammatory drugs (NSAIDs) on the ultrastructural aspects, uptake capacity, and biotransformation were investigated in green leafy vegetables. Thus, diclofenac (DCF), ibuprofen (IBU), and naproxen (NAP) were selected to assess the impact on three of the most consumed leafy vegetables: Atriplex patula L., Spinacia oleracea L., and Lactuca sativa L. The abiotic stress consisted of watering the vegetables once every 2 days, with different concentrations of the chosen NSAIDs (from 0.1 to 1 mg L−1). The uptake in leaves and soil was determined by high-performance liquid chromatography, after solid-phase extraction successfully optimized herein. The morphological and anatomical changes were monitored through scanning and transmission electron microscopy analyses. The concentration of the NSAIDs determined in the leaf samples increased in a dose-dependent manner and the analyzed soil samples had NSAIDs concentrations ranging from 0.38 μg g−1 soil to 9.00 μg g−1 soil. The morphological aspect of the green leafy vegetables was not affected compared to untreated controls, while several ultrastructural changes were observed, where oily accumulations were detected in the cell walls and vacuoles and damaged mitochondria with ruptured membranes were frequent. The obtained results demonstrated that under the current experimental abiotic stress conditions, edible plants can uptake high amounts of NSAIDs, depending on the concentration used. As a future perspective, this could also have adverse effects on the consumers through bioaccumulation or other metabolic processes.

Allelopathic root inhibition and its mechanisms

Allelopathy represents a valuable biochemical strategy in plant-plant interactions among different plants, e.g. among crops and weeds. It is an important strategy in the colonization of many invasive alien plants. Allelopathic plants affect the growth of other plants in the vicinity through the release of secondary metabolites (allelochemicals) into the soil. In particular, many allelochemicals suppress the root growth of target plants, but little is known about the mechanism involved in root growth inhibition. In this review, we will highlight the mechanism of root suppression involving: (i) Alterations in auxin homeostasis affecting polar auxin transport and root gravitropism, (ii) Biochemical and physiological processes in inhibited roots associated with oxidative stress due to direct production and accumulation of reactive oxygen species or suppression of antioxidative response and (iii) The ultrastructural modifications in root tip exposed to allelochemicals that drastically suppress the cell division and eventually lead to shorter roots of target plants.

Effect of oocyte vitrification on glucose transport in mouse metaphase II oocytes.

Oocyte vitrification has significantly improved the survival rate and become the mainstream method for cryopreserving oocytes. Previous studies have demonstrated that the ultrastructure, mitochondrial function, DNA methylation, and histone modification exhibit an irreversible effect after oocyte vitrification. However, little is known about the effects of oocyte vitrification on glucose transport and metabolism. This study aims to determine whether mouse oocyte vitrification causes abnormal glucose metabolism and identify a strategy to correct abnormal glucose metabolism. Furthermore, this study further investigates the effects of oocyte vitrification on glucose uptake, and glucose metabolism, and energy levels. The results indicated that vitrification significantly reduced the glucose transport activity, NADPH, glutathione, and ATP levels, and increased reactive oxygen species levels in oocytes (P < 0.01). Vitrification also reduced the expression of glucose transporter isoform 1 (GLUT1) (P < 0.01). Adding a GLUT1 inhibitor reduced the glucose uptake capacity of oocytes. Furthermore, the inclusion of vitamin C into thawing and culture solutions restored abnormal glucose transportation and metabolism and improved the survival, two-cell embryo, and blastocyst rates of the vitrified groups via parthenogenesis (P < 0.05). Overall, this method may improve the quality and efficiency of oocyte vitrification.

Nano-Scale Modifications of Amniotic Membrane Induced by UV and Antibiotic Treatment: Histological, AFM and FTIR Spectroscopy Evidence.

The efficiency of amniotic membrane (AM) transplantation in different types of ocular surface disorders is due to its outstanding properties such as antifibrotic, antibacterial, anti-inflammatory and antiangiogenic, working as a versatile scaffold to promote corneal tissue epithelialization. A proper preparation, preservation and clinical application are crucial for the best outcomes in the treatment of different severe ocular disorders, taking into account its fragility. In this context, by combining high-sensitivity tools such as atomic force microscopy (AFM) and Fourier transform infrared (FTIR) spectroscopy with histological and immunohistochemical examination, we aimed to investigate the ultrastructural modifications of the amniotic membrane (AM) upon UV exposure and/or antibiotic treatment, with relevance for clinical applications in ocular surface surgery. From the morphological point of view, we noticed a loss of cuboidal cells in the basal membrane, accompanied by the splitting of collagen fibers upon UV and/or gentamicin treatment, while structural alteration of proteins was evidenced by the FTIR quantitative analysis of the secondary structure. A decrease in α-helix and β-sheet content, accompanied by increased content in less ordered structures (turns, random and side chains), was noticed after all the treatments. At the nano-scale, AFM details showed modifications of collagen fibrils in terms of their thickness and network compaction upon gentamicin and/or UV treatment. The enzymatic digestion assay demonstrated that UV exposure significantly reduces the degradation rate of the AM, while gentamicin treatment promotes an accelerated enzymatic digestion upon UV exposure. In order to highlight the clinical impact of the research, a clinical case is presented showing the relevance of amniotic membrane transplantation in pterygium surgery.

From sperm plug formation to ovulation: morphological and ultrastructural modifications in the seminal receptacle of the blue crab Callinectes danae

In species of Callinectes a sperm plug is formed in the seminal receptacle (SR) during the sperm transfer, which hinders the insemination by other males. In this study, we evaluated the morphology, histochemistry, and ultrastructure of the seminal receptacle of juvenile and adult females of Callinectes danae along the different stages of ovarian development. We described the histochemical changes that take place inside the SR, from the sperm plug formation until fertilization. We also quantified the amount of seminal fluid received by the females during copulation. Callinectes danae has a dorsal-type SR, which is common in Portunidae. The sperm plug is formed by the orderly deposition of seminal fluid of the anterior and median vas deferens, which in turn influences the secretion of the posterior region and produces chemically distinct strata. The dissolution of the sperm plug inside the SR occurs in synchrony with the spermatophore dehiscence and ovarian development. A holocrine secretion from dense layer cells dissolves the sperm plug. These cells have electron-dense and electron-lucid vesicles. The plug is completely dissolved when the females are at the intermediary ovarian development. At this stage, the SR dense layer folds and forms the lateral chamber. The dorsal oviduct, continuous to the dense layer, moves to a ventral-lateral position opposed to the vagina. During ovulation, the oocytes of mature females pass through the oviduct and the fertilization occurs almost above the vagina. The male ejaculates all its seminal fluid in a single copulation, leading to a total sperm depletion. Therefore, the whole sperm plug is produced by the males, through the combined secretions from three vas deferens regions. We show, for the first time, that the secretions from the anterior vas deferens also participate in the sperm plug formation. The female dissolves the plug, which is a crucial morphological change of the SR during the processes of fertilization and egg release. In species of Callinectes there was a clear co-evolution between the male and female reproductive systems: the male invests all its reproductive potential to build the sperm plug, and the female, to dissolve it.

Chromium (VI)-Induced Leaf-Based Differential Physiological, Metabolic and Microstructural Changes in Two Transgenic Cotton Cultivars (J208, Z905) and Their Hybrid Line (ZD14)

Chromium-VI (Cr (VI)) is a toxic metal pollutant to both plants and animals. In this study, we cultured two transgenic cotton cultivars (J208 (herbicide-resistant), Z905 (insect-resistant)) and their hybrid line (ZD14) in hydroponic media using 0, 10, 50, 100 µM Cr/L for 7 days. Leaf-related growth parameters such as leaf length and width considerably declined. Decrease in leaves’ fresh biomass was less in Z905 followed by ZD14 and J208, while the leaves dry biomass was less in Z905 followed by J208 and ZD14, respectively. Tolerance potential of plant height, root dry weight and leaf dry weight of both transgenic cotton cultivars significantly decreased at Cr concentration (100 µM) as compared with their hybrid line. Cr accumulation in roots tissues was higher (> 90%) than leaf tissues. Stress biomarkers (i.e., (Malondialdehyde (MDA), hydrogen peroxide (H2O2), total soluble protein (TSP)) as a whole upregulated in leaf tissues of all experimental cultivars. Differential changes in superoxide dismutase (SOD) activity were observed, while peroxidase (POD), catalase (CAT) and glutathione-reductase (GR) activities almost showed increasing trend in all cultivars under exceeding levels of Cr. Correlation studies, in terms of principal component analysis (PCA), revealed that 75.36% of the total variation was present among all the traits. The leaf-based ultrastructural modifications at 100 µM Cr stress level occurred more in Z905 followed by J208 and ZD14. All these changes revealed differential role of experimental cotton cultivars against Cr stress. Hybrid line (ZD14) performed well in comparison with its parental cultivars (J208, Z905) as a whole. Deeper molecular studies like transcriptomic and metabolomic need to be carried out to explore the genetic facts regarding such differential responses among these cotton cultivars under Cr stress.

Modifications of Adrenal Gland Ultrastructure in Streptozotocin-Induced Diabetic Model Rats

Modifications of Adrenal Gland Ultrastructure in Streptozotocin-Induced Diabetic Model Rats

Ultrastructural modifications induced by SARS-CoV-2 in Vero cells: a kinetic analysis of viral factory formation, viral particle morphogenesis and virion release.

Many studies on SARS-CoV-2 have been performed over short-time scale, but few have focused on the ultrastructural characteristics of infected cells. We used TEM to perform kinetic analysis of the ultrastructure of SARS-CoV-2-infected cells. Early infection events were characterized by the presence of clusters of single-membrane vesicles and stacks of membrane containing nuclear pores called annulate lamellae (AL). A large network of host cell-derived organelles transformed into virus factories was subsequently observed in the cells. As previously described for other RNA viruses, these replication factories consisted of double-membrane vesicles (DMVs) located close to the nucleus. Viruses released at the cell surface by exocytosis harbored the typical crown of spike proteins, but viral particles without spikes were also observed in intracellular compartments, possibly reflecting incorrect assembly or a cell degradation process.

Transient ultrasound stimulation has lasting effects on neuronal excitability

BackgroundTranscranial ultrasound stimulation can acutely modulate brain activity, but the lasting effects on neurons are unknown. ObjectiveTo assess the excitability profile of neurons in the hours following transient ultrasound stimulation. MethodsPrimary rat cortical neurons were stimulated with a 40 s, 200 kHz pulsed ultrasound stimulation or sham-stimulation. Intrinsic firing properties were investigated through whole-cell patch-clamp recording by evoking action potentials in response to somatic current injection. Recordings were taken at set timepoints following ultrasound stimulation: 0–2 h, 6–8 h, 12–14 h and 24–26 h. Transmission electron microscopy was used to assess synaptic ultrastructure at the same timepoints. ResultsIn the 0–2 h window, neurons stimulated with ultrasound displayed an increase in the mean frequency of evoked action potentials of 32% above control cell levels (p = 0.023). After 4–6 h this increase was measured as 44% (p = 0.0043). By 12–14 h this effect was eliminated and remained absent 24–26 h post-stimulation. These changes to action potential firing occurred in conjunction with statistically significant differences between control and ultrasound-stimulated neurons in action potential half-width, depolarisation rate, and repolarisation rate, that were similarly eliminated by 24 h following stimulation. These effects occurred in the absence of alterations to intrinsic membrane properties or synaptic ultrastructure. ConclusionWe report that stimulating neurons with 40 s of ultrasound enhances their excitability for up to 8 h in conjunction with modifications to action potential kinetics. This occurs in the absence of major ultrastructural change or modification of intrinsic membrane properties. These results can inform the application of transcranial ultrasound in experimental and therapeutic settings.

Leaf structure and ultrastructure changes induced by heat stress and drought during seed filling in field-grown soybean and their relationship with grain yield.

Studies focusing on terminal drought combined with heat impacts on plants of agronomic value remain scarce, and even less under field conditions. The objective of this study was to investigate leaf structural and ultrastructural changes induced by heat stress (HS) and drought stress (DS) during seed filling and their relationship with physiological variables and yield determination. Two soybean cultivars were grown in field conditions. During seed filling four treatments were applied, including a control (without manipulation, at ambient temperature and field capacity), HS (episodes exceeding 32°C for 6 h d-1) during 21-d, DS (20% of field capacity soil water content) during 35-d, and HS×DS. Drought principally reduced leaf area, whereas heat decreased leaf thickness, possible as acclimation strategies, but also irreversible reducing CO2 assimilation sites. Both stresses damaged the outer and inner membranes of chloroplasts, causing swollen chloroplasts and accumulation of plastoglobules, loss of chlorophyll content, and negatively affecting chlorophyll fluorescence. Thus, the performance and integrity of the photosynthetic machinery were reduced. Through a morpho-functional perspective and a holistic multiscale approach, our results provide evidence of photosynthesis impairment and yield drops under stressful conditions which were associated with structural and ultrastructural (particularly at the level of chloroplasts) modifications of leaves.

Effect of the transgenerational exposure to elevated CO2 on low temperature tolerance of winter wheat: Chloroplast ultrastructure and carbohydrate metabolism

AbstractThe transgenerational effect of elevated atmospheric CO2 concentration (e[CO2]) on low temperature response in wheat is still little investigated, through the interaction of e[CO2], and low‐temperature stress has been reported in a single generation. Here, the low temperature‐induced modifications of chloroplast ultrastructure and carbohydrate metabolism in wheat after four generations continuously grown under ambient CO2 concentration (a[CO2]) and e[CO2] (2014–2018) were investigated. The results indicated that the transgenerational exposure to e[CO2] increased the number of grana lamellae and the amounts of osmiophilic lipid droplets, attenuating the negative effect of low temperature on chloroplast ultrastructure. The transgenerational e[CO2] enhanced the activities of antioxidant enzymes (i.e. SOD, POD and CAT) and concentrations of osmotic substances (i.e. proline and soluble sugar), which alleviated the low temperature‐induced oxidative damage to the chloroplast ultrastructure. In addition, transgenerational exposure of wheat to e[CO2] increased activities of vacInv and cwInv, while decreased fructokinase activity, which affected the sucrose metabolism in wheat leaf. These findings elucidated that transgenerational exposure to e[CO2] could improve low temperature tolerance of winter wheat, which provide novel insights to the response of wheat to future climate change.

Convergent Evolution of Broadband Reflectors Underlies Metallic Coloration in Butterflies

Butterfly wings often display structural colors, which are the result of light reflection from chitinous nanostructures that adorn the wing scales. Amongst these structural colors are broadband metallic reflections, which have been previously linked to an ultrathin broadband reflector in the nymphalid butterfly Argyrophorus argenteus. To test if similar optical modes of broadband, specular reflectance have evolved in other butterfly taxa, we characterized the reflective scales of eight species from five Papilionoidea families using microspectrophotometry (MSP), light microscopy in reflected and transmitted modes, and scanning electron microscopy (SEM). In Nymphalidae, Pieridae, and Hesperidae, and Lycaenidae, we find that broadband specularity is due to spatial mixing of densely juxtaposed colorful reflectances that change across microscale distances (e.g. 1-3 µm). These seemingly convergent silver scales are unpigmented, show a continuous upper lamina with reduced windows, and consist of an air-cuticle sandwich of variable thickness, forming an undulatory thin-film. Strikingly, Hypochrysops apelles (Lycaenidae) show a novel mode of silver reflectance with spatial color mixing occurring across the entire proximo-distal length of the scale (>100 µm), transitioning from blue to red hues between the stem and the tip of the scales. Unlike the undulatory type, this reflector shows flat thin-films but this includes a multilayered lower lamina responsible for selective color iridescence in other lycaenids or in sunset moths. Finally, the gold scales of Anteros formosus (Riodinidae) show mixed reflectance in the green-to-red range, seemingly produced by a thin film in the lower lamina. Our comparative study suggests that evolution of metallic broadband reflectance repeatedly involved spatial color mixing and unperforated upper laminae, and is accomplished using at least three types of ultrastructural modifications. Undulatory thin-film systems, based on geometric adjustments of the transverse profile of the upper lamina and scale lumen, are widespread and may have evolved repeatedly from more generic colorless scale morphologies, while lycaenid and riodinid broadband reflectors may be elaborations of pre-existing iridescent states.

Mode of action of a formulation containing hydrazones and saponins against leishmania spp. Role in mitochondria, proteases and reinfection process

Toxicity and poor adherence to treatment that favors the generation of resistance in the Leishmania parasites highlight the need to develop better alternatives. Here, we evaluated the in vitro effectiveness of hydrazone derived from chromanes 2-(2,3-dihydro-4H-1-benzothiopyran-4-ylidene) hydrazide (TC1) and 2-(2,3-dihydro-4H-1-benzopyran-4-ylidene) hydrazide (TC2) and the mixture of triterpene saponin hederagenin-3-O-(3,4-O-diacetyl-ß-D-xylopyranosyl-(1à3)-a-L- rhamnopyranosyl-(1à2)-a-L-arabinofuranoside, hederagenin-3-O-(3,4-O-diacetyl-a-L- arabinopyranosyl-(1à3)-a-L-rhamnopyranosyl-(1à2)-a-L-arabinofuranoside and, hederagenin-3-O-(4-O-acetyl-ß-D-xylopyranosyl-(1à3)-a-L-rhamnopyranosyl-(1à2)-a-L-arabinofuranoside from Sapindus saponaria (SS) on L. braziliensis and L. pifanoi. Mixtures of TC1 or TC2 with saponin were formulated for topical application and the therapeutic effectiveness was evaluated in the model for cutaneous leishmaniasis (CL) in golden hamster. The mode of action of these compounds was tested on various parasite processes and ultrastructural parasite modifications. TC1, TC2 and SS showed moderate cytotoxicity when tested independently but toxicity was improved when tested in combination. The compounds were more active against intracellular Leishmania amastigotes. In vivo studies showed that combinations of TC1 or TC2 with SS in 1:1 ratio (w/w) cured 100% of hamsters with no signs associated with toxicity. The compounds did cause changes in the mitochondrial activity of the parasite with a decrease in ATP levels and depolarization of membrane potential and overproduction of reactive oxygen species; nevertheless, these effects were not related to alterations in membrane permeability. The phagolysosome ultrastructure was also affected impacting the survival of Leishmania but the function of the lysosome nor the pH inside the phagolysosome did not change. Lastly, there was a protease inhibition which was directly related to the decrease in the ability of Leishmania to infect and multiply inside the macrophage. The results suggest that the combination of TC1 and TC2 with SS in a 1:1 ratio is capable of curing CL in hamsters. This effect may be due to the ability of these compounds to affect parasite survival and the ability to infect new cells.

Growth Temperature Influence on Lipids and Photosynthesis in Lepidium sativum.

Temperature has a major impact on plant development and growth. In temperate climates, the seasonal temperature displays large variations that can affect the early stages of plant growth and development. Sessile organisms need to be capable of responding to these conditions, so that growth temperature induces morphological and physiological changes in the plant. Besides development, there are also important molecular and ultrastructural modifications allowing to cope with different temperatures. The chloroplast plays a crucial role in plant energetic metabolism and harbors the photosynthetic apparatus. The photosynthetic light reactions are at the interface between external physical conditions (light, temperature) and the cell biochemistry. Therefore, photosynthesis requires structural flexibility to be able to optimize its efficiency according to the changes of the external conditions. To investigate the effect of growth temperature on the photosynthetic apparatus, we followed the photosynthetic performances and analyzed the protein and lipid profiles of Lepidium sativum (cress) grown at three different temperatures. This revealed that plants developing at temperatures above the optimum have a lower photosynthetic efficiency. Moreover, plants grown under elevated and low temperatures showed a different galactolipid profile, especially the amount of saturated galactolipids decreased at low temperature and increased at high temperature. From the analysis of the chlorophyll a fluorescence induction, we assessed the impact of growth temperature on the re-oxidation of plastoquinone, which is the lipidic electron carrier of the photosynthetic electron transport chain. We show that, at low temperature, along with an increase of unsaturated structural lipids and plastochromanol, there is an increase of the plastoquinone oxidation rate in the dark. These results emphasize the importance of the thylakoid membrane composition in preserving the photosynthetic apparatus under non-optimal temperatures.

Study on ultra-structural effects caused by Onion yellow dwarf virus infection in ‘Rossa di Tropea’ onion bulb by means of magnetic resonance imaging

‘Rossa di Tropea’ onion is a particular pink/red coloured onion cultivated in Calabria region (Southern Italy), representing one of the Italian most important vegetable crops granted with Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) trademarks. This local cultivar is characterised by a high nutraceutical compounds content showing anti-inflammatory, anti-cholesterol, anticancer and antioxidant properties. As all vegetable crops and Allium spp., ‘Rossa di Tropea’ onion is affected by several viruses. Among these, the species Onion yellow dwarf virus (OYDV, genus Potyvirus, family Potyviridae), represents the most limiting biotic stress, inducing severe symptoms. OYDV effect on tissues architecture in whole bulbs was investigated using magnetic resonance microimaging (MRI) technique, which allows the interior of samples to be imaged non-invasively and non-destructively and yields quantitative information on physico-chemical parameters describing water mobility (T1 and T2 relaxation times). The use of such tool allowed to determine how OYDV alters plant physiology by inducing water accumulation in bulb tissues as well as causing ultra-structural modifications of cell wall, highlighted by MRI. All these effects resulted in an increase of free water in plant tissues, and consequently relevant water losses during post-harvest storage, seriously affecting bulb quality, marketability and shelf life.

Adrenergic Signaling-Induced Ultrastructural Strengthening of Intercalated Discs via Plakoglobin Is Crucial for Positive Adhesiotropy in Murine Cardiomyocytes.

Intercalated discs (ICDs), which connect adjacent cardiomyocytes, are composed of desmosomes, adherens junctions (AJs) and gap junctions (GJs). Previous data demonstrated that adrenergic signaling enhances cardiac myocyte cohesion, referred to as positive adhesiotropy, via PKA-mediated phosphorylation of plakoglobin (PG). However, it was unclear whether positive adhesiotropy caused ultrastructural modifications of ICDs. Therefore, we further investigated the role of PG in adrenergic signaling-mediated ultrastructural changes in the ICD of cardiomyocytes. Quantitative transmission electron microscopy (TEM) analysis of ICD demonstrated that cAMP elevation caused significant elongation of area composita and thickening of the ICD plaque, paralleled by enhanced cardiomyocyte cohesion, in WT but not PG-deficient cardiomyocytes. STED microscopy analysis supported that cAMP elevation ex vivo enhanced overlap of desmoglein-2 (Dsg2) and N-cadherin (N-cad) staining in ICDs of WT but not PG-deficient cardiomyocytes. For dynamic analyses, we utilized HL-1 cardiomyocytes, in which cAMP elevation induced translocation of Dsg2 and PG but not of N-cad to cell junctions. Nevertheless, depletion of N-cad but not of Dsg2 resulted in a decrease in basal cell cohesion whereas positive adhesiotropy was abrogated in monolayers depleted for either Dsg2 or N-cad. In the WT mice, ultrastrutural changes observed after cAMP elevation were paralleled by phosphorylation of PG at serine 665. Our data demonstrate that in murine hearts adrenergic signaling enhanced N-cad and Dsg2 in the ICD paralleled by ultrastrutural strengthening of ICDs and that effects induced by positive adhesiotropy were strictly dependent on Pg.