electron transmission microscopy
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2021 ◽  
Vol 73 (4) ◽  
pp. 799-811
Author(s):  
N.V. Sollecito ◽  
R.N. Alves ◽  
M.E. Beletti ◽  
E.C.M. Pereira ◽  
M.S. Miranda ◽  
...  

ABSTRACT This study aimed to evaluate the ultrastructural morphometry of bovine embryos produced in vitro grown at different concentrations of antioxidants. After in vitro maturation and fertilization, the presumptive zygotes were assigned into five treatments. T1) without the addition of any antioxidants (negative control); T2) addition of 50μM/mL cysteamine; and T3, T4 and T5) adding 2.5μg/mL, 5.0μg/mL or 10.0μg/mL of the antioxidants derived from the oily extract from Lippia origanoides, respectively. On D7 of culture, the embryos in the blastocyst stage were fixed and prepared for electron transmission microscopy. These were evaluated for the proportion of cytoplasm-to-nucleus, cytoplasm-to-mitochondria, cytoplasm-to-vacuoles, cytoplasm-to-autophagic vacuoles and cytoplasm-to-lipid droplets. Blastocysts cultured in media containing oily extract of Lippia origanoides presented morphological characteristics such as high cell:mitochondria ratio and low cell:vacuoles and cell:autophagic vacuole ratio, possibly been morphological indicators of embryonic quality. Inner cell mass (ICM) from blastocysts cultured in media without any antioxidants had the highest cell:vacuole ratio. Similar results were found in the trophectoderm (TE) cells of blastocysts from treatment 2. Embryo culture media supplemented with antioxidants derived from Lippia origanoides oil produced embryos with a higher cytoplasmic proportion of organelles, such as mitochondria. Also, treatments without any antioxidants or with the addition of cysteamine presented cytoplasmic vacuolization, a characteristic related to production of poor-quality embryos.


2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Laura Pandolfi ◽  
Tommaso Fossali ◽  
Vanessa Frangipane ◽  
Sara Bozzini ◽  
Monica Morosini ◽  
...  

Abstract Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic proportions. Given that the main target of SARS-CoV-2 are lungs leading to severe pneumonia with hyperactivation of the inflammatory cascade, we conducted a prospective study to assess alveolar inflammatory status in patients with moderate to severe COVID-19. Methods Diagnostic bronchoalveolar lavage (BAL) was performed in 33 adult patients with SARS-CoV-2 infection by real-time PCR on nasopharyngeal swab admitted to the Intensive care unit (ICU) (n = 28) and to the Intermediate Medicine Ward (IMW) (n = 5). We analyze the differential cell count, ultrastructure of cells and Interleukin (IL)6, 8 and 10 levels. Results ICU patients showed a marked increase in neutrophils (1.24 × 105 ml− 1, 0.85–2.07), lower lymphocyte (0.97 × 105 ml− 1, 0.024–0.34) and macrophages fractions (0.43 × 105 ml− 1, 0.34–1.62) compared to IMW patients (0.095 × 105 ml− 1, 0.05–0.73; 0.47 × 105 ml− 1, 0.28–1.01 and 2.14 × 105 ml− 1, 1.17–3.01, respectively) (p < 0.01). Study of ICU patients BAL by electron transmission microscopy showed viral particles inside mononuclear cells confirmed by immunostaining with anti-viral capsid and spike antibodies. IL6 and IL8 were significantly higher in ICU patients than in IMW (IL6 p < 0.01, IL8 p < 0.0001), and also in patients who did not survive (IL6 p < 0.05, IL8 p = 0.05 vs. survivors). IL10 did not show a significant variation between groups. Dividing patients by treatment received, lower BAL concentrations of IL6 were found in patients treated with steroids as compared to those treated with tocilizumab (p < 0.1) or antivirals (p < 0.05). Conclusions Alveolitis, associated with COVID-19, is mainly sustained by innate effectors which showed features of extensive activation. The burden of pro-inflammatory cytokines IL6 and IL8 in the broncho-alveolar environment is associated with clinical outcome.


2020 ◽  
Author(s):  
Laura Pandolfi ◽  
Fossali Tommaso ◽  
Frangipane Vanessa ◽  
Bozzini Sara ◽  
Morosini Monica ◽  
...  

Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic proportions. Given that the main target of SARS-CoV-2 are lungs leading to severe pneumonia with hyperactivation of the inflammatory cascade, we conducted a prospective study to assess alveolar inflammatory status in patients with moderate to severe COVID-19.Methods: Diagnostic bronchoalveolar lavage (BAL) was performed in 33 adult patients with SARS-CoV-2 infection by real-time PCR on nasopharyngeal swab admitted to the Intensive care unit (ICU) (n=28) and to the Intermediate Medicine Ward (IMW) (n=5). We analyze the differential cell count, ultrastructure of cells and Interleukin(IL)6, 8 and 10 levels.Results: ICU patients showed a marked increase in neutrophils (1.24 x 105 ml-1 , 0.85-2.07), lower lymphocyte (0.97 x 105 ml-1, 0.024-0.34) and macrophages fractions (0.43 x 105 ml-1, 0.34-1.62) compared to IMW patients (0.095 x 105 ml-1, 0.05-0.73; 0.47 x 105 ml-1, 0.28-1.01 and 2.14 x 105 ml-1, 1.17-3.01, respectively) (p<0.01). Study of ICU patients BAL by electron transmission microscopy showed viral particles inside mononuclear cells confirmed by immunostaining with anti-viral capsid and spike antibodies. IL6 and IL8 were significantly higher in ICU patients than in IMW (IL6 p<0.01, IL8 p<0.0001), and also in patients who did not survive (IL6 p < 0.05, IL8 p = 0.05 vs. survivors). IL10 did not show a significant variation between groups. Dividing patients by treatment received, lower BAL concentrations of IL6 were found in patients treated with steroids as compared to those treated with tocilizumab (p<0.1) or antivirals (p<0.05).Conclusions: Alveolitis, associated with COVID-19, is mainly sustained by innate effectors which showed features of extensive activation. The burden of pro-inflammatory cytokines IL6 and IL8 in the broncho-alveolar environment is associated with clinical outcome.


2020 ◽  
Author(s):  
laura pandolfi ◽  
Fossali Tommaso ◽  
Frangipane Vanessa ◽  
Bozzini Sara ◽  
Morosini Monica ◽  
...  

Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic proportions. Given that the main target of SARS-CoV-2 are lungs leading to severe pneumonia with hyperactivation of the inflammatory cascade, we conducted a prospective study to assess alveolar inflammatory status in patients with moderate to severe COVID-19. Methods: Diagnostic bronchoalveolar lavage (BAL) was performed in 33 adult patients with SARS-CoV-2 infection by real-time PCR on nasopharyngeal swab admitted to the Intensive care unit (ICU) (n=28) and to the Intermediate Medicine Ward (IMW) (n=5). We analyze the differential cell count, ultrastructure of cells and Interleukin(IL)6, 8 and 10 levels.Results: ICU patients showed a marked increase in neutrophils (1.24 x 105 ml-1 , 0.85-2.07), lower lymphocyte (0.97 x 105 ml-1, 0.024-0.34) and macrophages fractions (0.43 x 105 ml-1, 0.34-1.62) compared to IMW patients (0.095 x 105 ml-1, 0.05-0.73; 0.47 x 105 ml-1, 0.28-1.01 and 2.14 x 105 ml-1, 1.17-3.01, respectively) (p<0.01). Study of ICU patients BAL by electron transmission microscopy showed viral particles inside mononuclear cells confirmed by immunostaining with anti-viral capsid and spike antibodies. IL6 and IL8 were significantly higher in ICU patients than in IMW (IL6 p<0.01, IL8 p<0.0001), and also in patients who did not survive (IL6 p < 0.05, IL8 p = 0.05 vs. survivors). IL10 did not show a significant variation between groups. Dividing patients by treatment received, lower BAL concentrations of IL6 were found in patients treated with steroids as compared to those treated with tocilizumab (p<0.1) or antivirals (p<0.05). Conclusions: Alveolitis, associated with COVID-19, is mainly sustained by innate effectors which showed features of extensive activation. The burden of pro-inflammatory cytokines IL6 and IL8 in the broncho-alveolar environment is associated with clinical outcome.


2020 ◽  
Author(s):  
laura pandolfi ◽  
Fossali Tommaso ◽  
Frangipane Vanessa ◽  
Bozzini Sara ◽  
Morosini Monica ◽  
...  

Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic proportions. Given that the main target of SARS-CoV-2 are lungs leading to severe pneumonia with hyperactivation of the inflammatory cascade, we conducted a prospective study to assess alveolar inflammatory status in patients with moderate to severe COVID-19. Methods: Diagnostic bronchoalveolar lavage (BAL) was performed in 33 adult patients with SARS-CoV-2 infection by real-time PCR on nasopharyngeal swab admitted to the Intensive care unit (ICU) (n=28) and to the Intermediate Medicine Ward (IMW) (n=5). We analyze the differential cell count, ultrastructure of cells and Interleukin(IL)6, 8 and 10 levels.Results: ICU patients showed a marked increase in neutrophils (1.24 x 105 ml-1 , 0.85-2.07), lower lymphocyte (0.97 x 105 ml-1, 0.024-0.34) and macrophages fractions (0.43 x 105 ml-1, 0.34-1.62) compared to IMW patients (0.095 x 105 ml-1, 0.05-0.73; 0.47 x 105 ml-1, 0.28-1.01 and 2.14 x 105 ml-1, 1.17-3.01, respectively) (p<0.01). Study of ICU patients BAL by electron transmission microscopy showed viral particles inside mononuclear cells confirmed by immunostaining with anti-viral capsid and spike antibodies. IL6 and IL8 were significantly higher in ICU patients than in IMW (IL6 p<0.01, IL8 p<0.0001), and also in patients who did not survive (IL6 p < 0.05, IL8 p = 0.05 vs. survivors). IL10 did not show a significant variation between groups. Dividing patients by treatment received, lower BAL concentrations of IL6 were found in patients treated with steroids as compared to those treated with tocilizumab (p<0.1) or antivirals (p<0.05). Conclusions: Alveolitis, associated with COVID-19, is mainly sustained by innate effectors which showed features of extensive activation. The burden of pro-inflammatory cytokines IL6 and IL8 in the broncho-alveolar environment is associated with clinical outcome.


Plants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1194
Author(s):  
Mikołaj Kostryco ◽  
Mirosława Chwil ◽  
Renata Matraszek-Gawron

The genus Rubus is one of the largest taxonomically diverse and complex genera in the family Rosaceae. Morphology of pollen grains (equatorial and polar axes length, shape and size, aperture position, exine sculpture, perforations) is regarded as one of its main diagnostic features for identification of species and varieties. An attempt was made to fill the gap concerning the pollen micromorphology and ultrastructure of R. idaeus L. using light, scanning, and electron transmission microscopy. This study is a comparative analysis of micromorphological and ultrastructural traits of pollen from six raspberry cultivars. The pollen grains were classified as small or medium of shape prolato-spheroids. The parallel striae in the equatorial view in the exine sculpture were sometimes branched dichotomously in ‘Glen Ample’, ‘Polka’, and ‘Polana’, arcuate in ‘Laszka’ and ‘Pokusa’, or irregularly overlapping in ‘Radziejowa’. The width of exine striae of biennial fruiting cultivars was much larger than in repeated fruiting cultivars. In terms of the increasing number of perforations per unit area of the exine surface, the cultivars were ranked as follows: ‘Pokusa’ < ‘Glen Ample’ < ‘Laszka’ < ‘Polka’ < ‘Polana’ < ‘Radziejowa’. The thickest tectum, the highest and thickest columellae with the largest distances between them, and the thicker foot layer were demonstrated in ‘Glen Ample’. The ectoexine constituted on average ca. 78–90% of the exine thickness. The findings may constitute auxiliary traits i.a. for identification of related taxa, interpretation of phylogenetic relationships, and pollination biology.


2018 ◽  
Vol 15 (29) ◽  
pp. 266-275
Author(s):  
F. PILAQUINGA ◽  
V. ESCOBAR ◽  
G. CUEVA ◽  
L. MENESES

The green synthesis of silver nanoparticles aims to synergistically bind bactericidal properties of the element with the chemical composition of plant extracts and potentially use them in biodegradable plastic substitute materials. In this work, a degradable gelatin film was prepared with silver nanoparticles synthesized with aqueous extract of garlic (Allium sativum); the nanoparticles were previously characterized by ultraviolet-visible spectrophotometry, electron scanning microscopy and electron transmission microscopy. To make the film, pure gelatin, gelatin - carboxymethylcellulose - glycerol, and gelatin - arabic gum mixtures were tested, obtaining the desired texture only with gelatin, at different concentrations. The film with the greatest flexibility was 10% gelatin, with puncture resistance of 47 N and density of 0.943 g/mL. This film was analyzed by infrared spectroscopy, finding modifications of the spectrum dependent on the presence of silver nanoparticles-garlic extract. The electron scanning microscopy revealed a uniform distribution of silver in the film (74.4nm) with an abundance of 56.20% and a concentration of 1.58 mg/g determined by atomic absorption spectrophotometry. Finally, the prepared gelatin film is resistant, amber colored, with possible bactericidal properties due to the presence of silver nanoparticles and easily degradable at 50°C in water.


2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
L. G. Rangel-Chávez ◽  
M. I. Neria-González ◽  
A. Márquez-Herrera ◽  
M. Zapata-Torres ◽  
E. Campos-González ◽  
...  

A novel methodology for the direct synthesis of CdS nanoparticles, using a biological agent that avoids the extracellular processing, and the results of the characterization of CdS nanocrystals are presented. The by-products of the anaerobic respiratory process ofDesulfovibrio alaskensis6SR along with aqueous solutions of Cd salts were successfully employed to produce CdS nanocrystals with mixed cubic and hexagonal phases. Nanocrystal size has a narrow size distribution with little dependence on the Cd concentration. Both the presence of the crystallographic cubic phase and the crystalline order decrease as Cd concentration increases. The band gap values obtained from optical transmission measurements are lower than those of the bulk crystal. Raman spectroscopy characterization agrees with electron transmission microscopy images and X-ray diffraction results indicating that the method promotes the formation of high structural quality nanocrystals when low concentrations of the Cd salt are used.


2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Mihaela Birdeanu ◽  
Mirela Vaida ◽  
Eugenia Fagadar-Cosma

The present study is focused on the obtaining of the Zn3Nb2O8nanomaterial using the hydrothermal method and its characterization through different techniques. X-ray diffraction at room temperature revealed that a novel crystalline form of the nanomaterial forms at 1100°C belonging to monoclinic space group C2/c. Field-emission scanning electron microscopy evidenced the columnar morphology of the particle’s agglomeration and the high resolution electron transmission microscopy confirms the measured interplanar distances calculated from the X-ray diffraction experiments. Using the UV-VIS spectrum and Kubelka-Munk equations, the absorbance and the band gap for the Zn3Nb2O8nanomaterial were calculated. PL spectrum reveals a single peak at 465 nm corresponding to the blue color fluorescence. The novel crystalline nanomaterial might find applications in fluorescence covering of technical devices, due to its capacity to preserve blue fluorescence both in acrylic based paint and after embedding in isopropyl alcohol.


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