scholarly journals Localization and Absolute Quantification of Dopamine in Discrete Intravesicular Compartments Using NanoSIMS Imaging

2021 ◽  
Vol 23 (1) ◽  
pp. 160
Author(s):  
Stefania Rabasco ◽  
Tho D. K. Nguyen ◽  
Chaoyi Gu ◽  
Michael E. Kurczy ◽  
Nhu T. N. Phan ◽  
...  
Keyword(s):  
Secondary Ion Mass Spectrometry ◽  
Chemical Species ◽  
High Sensitivity ◽  
Absolute Quantification ◽  
Dense Core ◽  
Cellular Processes ◽  
Large Dense Core Vesicles ◽  
Model Cell ◽  
The Absolute ◽  
The Individual

The absolute concentration and the compartmentalization of analytes in cells and organelles are crucial parameters in the development of drugs and drug delivery systems, as well as in the fundamental understanding of many cellular processes. Nanoscale secondary ion mass spectrometry (NanoSIMS) imaging is a powerful technique which allows subcellular localization of chemical species with high spatial and mass resolution, and high sensitivity. In this study, we combined NanoSIMS imaging with spatial oversampling with transmission electron microscopy (TEM) imaging to discern the compartments (dense core and halo) of large dense core vesicles in a model cell line used to study exocytosis, and to localize 13C dopamine enrichment following 4–6 h of 150 μM 13C L-3,4-dihydroxyphenylalanine (L-DOPA) incubation. In addition, the absolute concentrations of 13C dopamine in distinct vesicle domains as well as in entire single vesicles were quantified and validated by comparison to electrochemical data. We found concentrations of 87.5 mM, 16.0 mM and 39.5 mM for the dense core, halo and the whole vesicle, respectively. This approach adds to the potential of using combined TEM and NanoSIMS imaging to perform absolute quantification and directly measure the individual contents of nanometer-scale organelles.

Relationship between golgi apparatus and axonal reticulum in sympathetic ganglion cells

1978 ◽  
Vol 36 (2) ◽  
pp. 598-599
Author(s):  
J. Quatacker ◽  
W. De Potter
Keyword(s):  
Golgi Apparatus ◽  
Sympathetic Ganglion ◽  
Phosphotungstic Acid ◽  
Ganglion Cells ◽  
Low Ph ◽  
Dense Core ◽  
Dense Core Vesicles ◽  
Large Dense Core Vesicles ◽  
Cervical Ganglia ◽  
Axoplasmic Reticulum

Mucopolysaccharides have been demonstrated biochemically in catecholamine-containing subcellular particles in different rat, cat and ox tissues. As catecholamine-containing granules seem to arise from the Golgi apparatus and some also from the axoplasmic reticulum we examined wether carbohydrate macromolecules could be detected in the small and large dense core vesicles and in structures related to them. To this purpose superior cervical ganglia and irises from rabbit and cat and coeliac ganglia and their axons from dog were subjected to the chromaffin reaction to show the distribution of catecholamine-containing granules. Some material was also embedded in glycolmethacrylate (GMA) and stained with phosphotungstic acid (PTA) at low pH for the detection of carbohydrate macromolecules.The chromaffin reaction in the perikarya reveals mainly large dense core vesicles, but in the axon hillock, the axons and the terminals, the small dense core vesicles are more prominent. In the axons the small granules are sometimes seen inside a reticular network (fig. 1).

scholarly journals Application of Vibrational Spectroscopies in the Qualitative Analysis of Gingival Crevicular Fluid and Periodontal Ligament during Orthodontic Tooth Movement

2021 ◽  
Vol 10 (7) ◽  
pp. 1405
Author(s):  
Fabrizia d’Apuzzo ◽  
Ludovica Nucci ◽  
Ines Delfino ◽  
Marianna Portaccio ◽  
Giuseppe Minervini ◽  
...  
Keyword(s):  
Periodontal Ligament ◽  
Tooth Movement ◽  
Gingival Crevicular Fluid ◽  
Orthodontic Tooth Movement ◽  
High Sensitivity ◽  
Data Analysis Procedure ◽  
Vibrational Spectroscopies ◽  
Composition Structure ◽  
The Individual ◽  
Crevicular Fluid

Optical vibrational techniques show a high potentiality in many biomedical fields for their characteristics of high sensitivity in revealing detailed information on composition, structure, and molecular interaction with reduced analysis time. In the last years, we have used these techniques for investigating gingival crevicular fluid (GCF) and periodontal ligament (PDL) during orthodontic tooth treatment. The analysis with Raman and infrared signals of GCF and PDL samples highlighted that different days of orthodontic force application causes modifications in the molecular secondary structure at specific wavenumbers related to the Amide I, Amide III, CH deformation, and CH3/CH2. In the present review, we report the most relevant results and a brief description of the experimental techniques and data analysis procedure in order to evidence that the vibrational spectroscopies could be a potential useful tool for an immediate monitoring of the individual patient’s response to the orthodontic tooth movement, aiming to more personalized treatment reducing any side effects.

scholarly journals HID-1 controls formation of large dense core vesicles by influencing cargo sorting and trans-Golgi network acidification

2017 ◽  
Vol 28 (26) ◽  
pp. 3870-3880 ◽  
Author(s):  
Blake H. Hummer ◽  
Noah F. de Leeuw ◽  
Christian Burns ◽  
Lan Chen ◽  
Matthew S. Joens ◽  
...  
Keyword(s):  
Biochemical Properties ◽  
Neuroendocrine Cells ◽  
Dense Core ◽  
Core Formation ◽  
Dense Core Vesicles ◽  
Atpase Subunit ◽  
Trans Golgi Network ◽  
Large Dense Core Vesicles ◽  
Golgi Network ◽  
Cargo Sorting

Large dense core vesicles (LDCVs) mediate the regulated release of neuropeptides and peptide hormones. They form at the trans-Golgi network (TGN), where their soluble content aggregates to form a dense core, but the mechanisms controlling biogenesis are still not completely understood. Recent studies have implicated the peripheral membrane protein HID-1 in neuropeptide sorting and insulin secretion. Using CRISPR/Cas9, we generated HID-1 KO rat neuroendocrine cells, and we show that the absence of HID-1 results in specific defects in peptide hormone and monoamine storage and regulated secretion. Loss of HID-1 causes a reduction in the number of LDCVs and affects their morphology and biochemical properties, due to impaired cargo sorting and dense core formation. HID-1 KO cells also exhibit defects in TGN acidification together with mislocalization of the Golgi-enriched vacuolar H+-ATPase subunit isoform a2. We propose that HID-1 influences early steps in LDCV formation by controlling dense core formation at the TGN.

Synthesis of Cu2O and ZnO Nanowires and their Heterojunction Nanowires by Thermal Evaporation: A Short Review

2014 ◽  
Vol 71 (5) ◽  
Author(s):  
Muhammad Arif Khan ◽  
Samsudi Sakrani ◽  
Syahida Suhaima ◽  
Yussof Wahab ◽  
Rosnita Muhammad
Keyword(s):  
Physical Vapor Deposition ◽  
Thermal Evaporation ◽  
Chemical Properties ◽  
High Sensitivity ◽  
Short Review ◽  
Zno Nanowires ◽  
Oxide Semiconductor ◽  
The Individual ◽  
Photovoltaic Solar Cells ◽  
Physical And Chemical

One dimensional metal oxide semiconductor nanowires of copper (I) oxide (Cu2O), zinc oxide (ZnO), and their heterojunction nanowires possess remarkable physical and chemical properties. ZnO and Cu2O areattractive because the metals are abundant on earth, inexpensive, nontoxic.Moreover, these oxides have useful optical and electrical properties suitable for a wide variety of electrical devices, because their electrical conduction can be predictably controlled by doping. We here restrict the disscussion using a Hot Tube Vacuum Thermal Evaporation. The NWs in these devices will be studied by physical vapor deposition known as vapor-liquid-solid (VLS). Therefore, we explore conventional methods, particularly the VLS of growing ZnO and Cu2O nanowires which are assisted by the catalyst.  In this short review, we report the individual and combined (Cu2O/ZnO) junction nanowires by PVD method.  The main advantages of these composite nanowires are the natural p-n characteristics, the broad light absorption, the high sensitivity to humidity changes, and the fast dynamic response. The combination of all characteristics offered by Cu2O/ZnO nanowires can enable the fabrication of diverse sensing devices, and photovoltaic solar cells.

scholarly journals Failure to collectively assess security surveillance technologies will inevitably lead to an absolute surveillance society.

2013 ◽  
Vol 12 (1) ◽  
pp. 73-88 ◽  
Author(s):  
Timothy Mitchener-Nissen
Keyword(s):  
New Technology ◽  
Security Technology ◽  
Security Technologies ◽  
Small Rise ◽  
The Absolute ◽  
Waking Up ◽  
Surveillance Society ◽  
The Individual ◽  
Security Surveillance ◽  
The Impact

When assessing any security technology which impacts upon privacy, whether this constitutes a new technology or the novel application of existing technologies, we should do so by examining the combined effect of all security interventions currently employed within a society. This contrasts with the prevailing system whereby the impact of a new security technology is predominantly assessed on an individual basis by a subjective balancing of the security benefits of that technology against any reductions in concomitant rights, such as privacy and liberty. I contend that by continuing to focus on the individual effect, as opposed to the combined effects, of security technologies within a society the likelihood of sleep-walking into (or indeed waking-up in) an absolute surveillance society moves from a possible future to the logically inevitable future. This conclusion is based on two underlying assertions. Firstly that assessing a technology often entails a judgement of whether any loss in privacy is legitimised by a justifiable increase in security; however one fundamental difference between these two rights is that privacy is a finite resource with identifiable end-states (i.e. absolute privacy through to the absolute absence of privacy) whereas security does not have two finite end-states (while there exists the absolute absence of security, absolute security is an unobtainable yet desired goal). The second assertion, which relies upon the validity of the first, holds that one consequence of absolute security being unobtainable yet desirable is that new security interventions will continuously be developed, each potentially trading a small measure of privacy for a small rise in security. Examined individually each intervention may constitute a justifiable trade-off. However this approach of combining interventions in the search for ever greater security will ultimately reduce privacy to zero.

scholarly journals Hegelian Phenomenology of Spirit and Boxing Fight

2018 ◽  
Vol 77 (1) ◽  
pp. 48-54
Author(s):  
Jerzy Kosiewicz
Keyword(s):  
The Body ◽  
Special Role ◽  
Collective Consciousness ◽  
Phenomenology Of Spirit ◽  
Human Individual ◽  
The Subject ◽  
The Absolute ◽  
Low Culture ◽  
The Individual ◽  
Absolute Synthesis

Abstract In the presented text the author points out to anthropological as well as axiological foundations of the boxing fight from the viewpoint of Hegel’s philosophy. In the genial idealist’s views it is possible to perceive the appreciation of the body, which constitutes a necessary basis for the man’s physical activity, for his work oriented towards the self-transformation and the transformation of the external world, as well as for rivalry and the hand-to-hand fight. While focusing our attention on the issue of rivalry and on the situation of the fight - and regarding it from the viewpoint of the master - slave theory (included in the phenomenology of spirit), it is possible to proclaim that even a conventionalised boxing fight - that is, restricted by cultural and sports rules of the game - has features of the fight to the death between two Hegelian forms of selfknowledge striving for self-affirmation and self-realisation. In the boxing fight, similarly as in the above mentioned Hegelian theory, a problem of work and of the development of the human individual (that is, of the subject, self-knowledge, the participant of the fight) appears. There appears also a prospect of death as a possible end of merciless rivalry. The fight revalues the human way in an important way, whereas the prospect for death, the awareness of its proximity, the feeling that its close and possible, saturates the life with additional values. It places the boxer, just like every subject fighting in a similar or a different way, on the path towards absolute abstraction - that is, it brings him closer to his self-fulfilment in the Absolute, to the absolute synthesis. The Hegelian viewpoint enables also to appreciate the boxing fight as a manifestation of low culture (being in contrast with high culture), to turn attention to the relations which - according to Hegel - take place between the Absolute and the man, as well as to show which place is occupied by the subject both in the process of the Absolute’s self-realisation and in the German thinker’s philosophical system. Independently of the dialectical, simultaneously pessimistic and optimistic overtone of considerations connected with the very boxing fight (regarding destruction and spiritualisation on a higher level), it is possible to perceive farreaching appreciation of the human individual in Hegel’s philosophy since the Absolute cannot make its own self-affirmation without the individual, without the human body, without the fight aimed at the destruction of the enemy and without the subjective consciousness and the collective consciousness which appear thanks to this fight. Thus, it is justified to suppose that the foundation of the whole Hegel’s philosophy is constituted by anthropology and that in the framework of this anthropology a special role is played by the fight and by work, which changes the subject and his(her) environment. Admittedly Hegel does not emphasise it explicitly, nevertheless his views (with their centre, which, according to Hegel himself and his interpreters, is constituted by the Absolute) have, as a matter of fact, an anthropocentric character and the main source of the subject’s development is the struggle which, irrespectively of its result, always primarily leads to the destruction or even to the death of one of the sides, just like in the boxing fight. However, it is also a germ of the positive re-orientation of the subject, the beginning and a continuation of that what the phenomenology of the spirit describes as a movement towards absolute abstraction.

scholarly journals The Complementary Role of ToF-SIMS in the Assessment of Imiquimod Permeated into the Skin from a Microemulsion Dosage Form

2019 ◽  
pp. 196-210
Keyword(s):  
Dosage Form ◽  
Secondary Ion Mass Spectrometry ◽  
Imaging Techniques ◽  
Skin Permeation ◽  
High Sensitivity ◽  
Tof Sims ◽  
Complementary Role ◽  
Nodular Lesions ◽  
Superficial Basal Cell Carcinoma ◽  
Fluorescent Tags

The assessment of drug permeation into/across the skin is traditionally accomplished using Franz diffusion cells with subsequent analysis by conventional chromatographic methods such as HPLC and more recently using advanced imaging techniques. In this context, time of flight-secondary ion mass spectrometry (ToF-SIMS) offers distinctive advantages in mapping drugs within skin with high sensitivity and chemical specificity without the need for fluorescent tags or radiolabels. The work in this paper uses the combination of conventional and advanced methods to evaluate imiquimod permeation into the skin. This approach provides complementary and detailed information regarding the permeated mass, the permeation depth and the spatial distribution and localisation of drugs within skin. Imiquimod is an immune modulator drug approved by the FDA for the treatment of superficial basal cell carcinoma (BCC) but not the nodular lesions. As other studies have reported that Aldara™ cream (imiquimod 5% w/w) has some limitations in the treatment of nodular BCC lesions due to the cream’s inability to deliver imiquimod into the deeper more invasive nodular lesions, an enhancement of imiquimod permeation is thought to be useful to overcome these limitations. Therefore, an attempt to improve delivery of imiquimod into the deeper skin layers using microemulsions was investigated. Imiquimod microemulsions were formulated and characterised in our previous work are now tested for skin permeation enhancement. However, the assessment of imiquimod permeation from the formulated microemulsions using HPLC and ToF-SIMS demonstrated a limited ability of the microemulsions to improve delivery of imiquimod over Aldara™ cream. This was attributed to the poor release of imiquimod from the microemulsion formulas due to the high affinity of imiquimod for the oil phase and the encapsulation of the oil droplets by the S/Co-S mixture. This is thought to be, the first time that ToF-SIMS has been used to assess permeation of imiquimod from a microemulsion dosage form.

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