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Article ; Online: Three-dimensional SEM, TEM, and STEM for analysis of large-scale biological systems.

Radulović, Snježana / Sunkara, Sowmya / Rachel, Reinhard / Leitinger, Gerd

2022 Volume 158, Issue 3, Page(s) 203–211

Abstract: A major aim in structural cell biology is to analyze intact cells in three dimensions, visualize subcellular structures, and even localize proteins at the best possible resolution in three dimensions. Though recently developed electron microscopy tools ... ...

Abstract	A major aim in structural cell biology is to analyze intact cells in three dimensions, visualize subcellular structures, and even localize proteins at the best possible resolution in three dimensions. Though recently developed electron microscopy tools such as electron tomography, or three-dimensional (3D) scanning electron microscopy, offer great resolution in three dimensions, the challenge is that, the better the resolution, usually the smaller the volume under investigation. Several different approaches to overcome this challenge were presented at the Microscopy Conference in Vienna in 2021. These tools include array tomography, batch tomography, or scanning transmission electron tomography, all of which can nowadays be extended toward correlative light and electron tomography, with greatly increased 3D information. Here, we review these tools, describe the underlying procedures, and discuss their advantages and limits.
MeSH term(s)	Electron Microscope Tomography/methods ; Imaging, Three-Dimensional/methods ; Microscopy, Electron, Scanning ; Microscopy, Electron, Scanning Transmission
Language	English
Publishing date	2022-07-12
Publishing country	Germany
Document type	Journal Article ; Review
ZDB-ID	1222930-1
ISSN	1432-119X ; 0301-5564 ; 0948-6143
ISSN (online)	1432-119X
ISSN	0301-5564 ; 0948-6143
DOI	10.1007/s00418-022-02117-w
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Article: Western diet-induced ultrastructural changes in mouse pancreatic acinar cells.

Lipovšek, Saška / Dolenšek, Jurij / Dariš, Barbara / Valladolid-Acebes, Ismael / Vajs, Tanja / Leitinger, Gerd / Stožer, Andraž / Skelin Klemen, Maša

Frontiers in cell and developmental biology

2024 Volume 12, Page(s) 1380564

Abstract: Mouse models of diet-induced type 2 diabetes mellitus provide powerful tools for studying the structural and physiological changes that are related to the disease progression. In this study, diabetic-like glucose dysregulation was induced in mice by ... ...

Abstract	Mouse models of diet-induced type 2 diabetes mellitus provide powerful tools for studying the structural and physiological changes that are related to the disease progression. In this study, diabetic-like glucose dysregulation was induced in mice by feeding them a western diet, and light and transmission electron microscopy were used to study the ultrastructural changes in the pancreatic acinar cells. Acinar necrosis and vacuolization of the cytoplasm were the most prominent features. Furthermore, we observed intracellular and extracellular accumulation of lipid compounds in the form of lipid droplets, structural enlargement of the cisternae of the rough endoplasmic reticulum (RER), and altered mitochondrial morphology, with mitochondria lacking the typical organization of the inner membrane. Last, autophagic structures, i.e., autophagosomes, autolysosomes, and residual bodies, were abundant within the acinar cells of western diet-fed mice, and the autolysosomes contained lipids and material of varying electron density. While diets inducing obesity and type 2 diabetes are clearly associated with structural changes and dysfunction of the endocrine pancreas, we here demonstrate the strong effect of dietary intervention on the structure of acinar cells in the exocrine part of the organ before detectable changes in plasma amylase activity, which may help us better understand the development of non-alcoholic fatty pancreas disease and its association with endo- and exocrine dysfunction.
Language	English
Publishing date	2024-03-14
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2024.1380564
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Article ; Online: Malpighian tubules in harvestmen.

Lipovšek, Saška / Kozel, Peter / Leitinger, Gerd / Novak, Tone

Protoplasma

2021 Volume 258, Issue 5, Page(s) 1145–1153

Abstract: In arachnids, the Malpighian tubules (MTs), coxal glands and stercoral pockets are capable of collecting and removing excreta from the body. The presence of the MTs among Opiliones was evidenced for the first time in Amilenus aurantiacus in 2015. ... ...

Abstract	In arachnids, the Malpighian tubules (MTs), coxal glands and stercoral pockets are capable of collecting and removing excreta from the body. The presence of the MTs among Opiliones was evidenced for the first time in Amilenus aurantiacus in 2015. Individuals undergo a winter diapause subterranean habitats. Here, we provided the morphological and cytological description of the MTs and asked whether their structure and ultrastructure change during the winter diapause. We studied the changes using light and transmission electron microscopy. The MTs consisted of the ureter and a pair of long, lateral blind-ended tubules, forming a long loop in the opisthosoma, and a coiled, terminal ball in the prosoma. The MTs were uniform, composed of a single-cell type, a monolayer of cuboidal epithelial cells, and the basal lamina. The cell ultrastructure was quite comparable to those in other arthropods, except for very long infoldings of the basal membrane protruding close to the nucleus. Except for spherite exploitation, no changes were observed in the ultrastructure of the MT epithelial cells during overwintering. We suggest that the analogous MTs in A. aurantiacus, and the nephron anatomies, along with a single-cell-type MT epithelium, might be of advantage in modelled studies of the nephron.
MeSH term(s)	Animals ; Ecosystem ; Epithelial Cells ; Humans ; Malpighian Tubules ; Microscopy, Electron, Transmission ; Seasons
Language	English
Publishing date	2021-03-29
Publishing country	Austria
Document type	Journal Article
ZDB-ID	123809-7
ISSN	1615-6102 ; 0033-183X
ISSN (online)	1615-6102
ISSN	0033-183X
DOI	10.1007/s00709-021-01634-0
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Article ; Online: Digging Deeper: Advancements in Visualization of Inhibitory Synapses in Neurodegenerative Disorders.

Radulović, Snježana / Sunkara, Sowmya / Maurer, Christa / Leitinger, Gerd

International journal of molecular sciences

2021 Volume 22, Issue 22

Abstract: Recent research has provided strong evidence that neurodegeneration may develop from an imbalance between synaptic structural components in the brain. Lately, inhibitory synapses communicating via the neurotransmitters GABA or glycine have come to the ... ...

Abstract	Recent research has provided strong evidence that neurodegeneration may develop from an imbalance between synaptic structural components in the brain. Lately, inhibitory synapses communicating via the neurotransmitters GABA or glycine have come to the center of attention. Increasing evidence suggests that imbalance in the structural composition of inhibitory synapses affect deeply the ability of neurons to communicate effectively over synaptic connections. Progressive failure of synaptic plasticity and memory are thus hallmarks of neurodegenerative diseases. In order to prove that structural changes at synapses contribute to neurodegeneration, we need to visualize single-molecule interactions at synaptic sites in an exact spatial and time frame. This visualization has been restricted in terms of spatial and temporal resolution. New developments in electron microscopy and super-resolution microscopy have improved spatial and time resolution tremendously, opening up numerous possibilities. Here we critically review current and recently developed methods for high-resolution visualization of inhibitory synapses in the context of neurodegenerative diseases. We present advantages, strengths, weaknesses, and current limitations for selected methods in research, as well as present a future perspective. A range of new options has become available that will soon help understand the involvement of inhibitory synapses in neurodegenerative disorders.
MeSH term(s)	Alzheimer Disease/diagnostic imaging ; Alzheimer Disease/drug therapy ; Alzheimer Disease/metabolism ; Alzheimer Disease/pathology ; Amyotrophic Lateral Sclerosis/diagnostic imaging ; Amyotrophic Lateral Sclerosis/drug therapy ; Amyotrophic Lateral Sclerosis/metabolism ; Amyotrophic Lateral Sclerosis/pathology ; Animals ; Brain/diagnostic imaging ; Brain/drug effects ; Brain/metabolism ; Brain/pathology ; Humans ; Huntington Disease/diagnostic imaging ; Huntington Disease/drug therapy ; Huntington Disease/metabolism ; Huntington Disease/pathology ; Levodopa/therapeutic use ; Memantine/therapeutic use ; Microscopy, Electron/methods ; Multiple Sclerosis/diagnostic imaging ; Multiple Sclerosis/drug therapy ; Multiple Sclerosis/metabolism ; Multiple Sclerosis/pathology ; Neuronal Plasticity/drug effects ; Neurons/drug effects ; Neurons/metabolism ; Neurons/ultrastructure ; Neuroprotective Agents/therapeutic use ; Neurotransmitter Agents/metabolism ; Parkinson Disease/diagnostic imaging ; Parkinson Disease/drug therapy ; Parkinson Disease/metabolism ; Parkinson Disease/pathology ; Synapses/drug effects ; Synapses/metabolism ; Synapses/ultrastructure ; Synaptic Transmission/drug effects ; Tetrabenazine/therapeutic use
Chemical Substances	Neuroprotective Agents ; Neurotransmitter Agents ; Levodopa (46627O600J) ; Memantine (W8O17SJF3T) ; Tetrabenazine (Z9O08YRN8O)
Language	English
Publishing date	2021-11-18
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms222212470
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Article: Malpighian tubules in harvestmen

Lipovšek, Saška / Kozel, Peter / Leitinger, Gerd / Novak, Tone

Protoplasma. 2021 Sept., v. 258, no. 5

2021

Abstract	In arachnids, the Malpighian tubules (MTs), coxal glands and stercoral pockets are capable of collecting and removing excreta from the body. The presence of the MTs among Opiliones was evidenced for the first time in Amilenus aurantiacus in 2015. Individuals undergo a winter diapause subterranean habitats. Here, we provided the morphological and cytological description of the MTs and asked whether their structure and ultrastructure change during the winter diapause. We studied the changes using light and transmission electron microscopy. The MTs consisted of the ureter and a pair of long, lateral blind-ended tubules, forming a long loop in the opisthosoma, and a coiled, terminal ball in the prosoma. The MTs were uniform, composed of a single-cell type, a monolayer of cuboidal epithelial cells, and the basal lamina. The cell ultrastructure was quite comparable to those in other arthropods, except for very long infoldings of the basal membrane protruding close to the nucleus. Except for spherite exploitation, no changes were observed in the ultrastructure of the MT epithelial cells during overwintering. We suggest that the analogous MTs in A. aurantiacus, and the nephron anatomies, along with a single-cell-type MT epithelium, might be of advantage in modelled studies of the nephron.
Keywords	Opiliones ; diapause ; epithelium ; excreta ; overwintering ; transmission electron microscopy ; ultrastructure ; ureter
Language	English
Dates of publication	2021-09
Size	p. 1145-1153.
Publishing place	Springer Vienna
Document type	Article
ZDB-ID	123809-7
ISSN	1615-6102 ; 0033-183X
ISSN (online)	1615-6102
ISSN	0033-183X
DOI	10.1007/s00709-021-01634-0
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Article: Investigation of the Influence of Wound-Treatment-Relevant Buffer Systems on the Colloidal and Optical Properties of Gold Nanoparticles.

Selmani, Atiđa / Jeitler, Ramona / Auinger, Michael / Tetyczka, Carolin / Banzer, Peter / Kantor, Brian / Leitinger, Gerd / Roblegg, Eva

Nanomaterials (Basel, Switzerland)

2023 Volume 13, Issue 12

Abstract: Biocompatible gold nanoparticles (AuNPs) are used in wound healing due to their radical scavenging activity. They shorten wound healing time by, for example, improving re-epithelialization and promoting the formation of new connective tissue. Another ... ...

Abstract	Biocompatible gold nanoparticles (AuNPs) are used in wound healing due to their radical scavenging activity. They shorten wound healing time by, for example, improving re-epithelialization and promoting the formation of new connective tissue. Another approach that promotes wound healing through cell proliferation while inhibiting bacterial growth is an acidic microenvironment, which can be achieved with acid-forming buffers. Accordingly, a combination of these two approaches appears promising and is the focus of the present study. Here, 18 nm and 56 nm gold NP (Au) were prepared with Turkevich reduction synthesis using design-of-experiments methodology, and the influence of pH and ionic strength on their behaviour was investigated. The citrate buffer had a pronounced effect on the stability of AuNPs due to the more complex intermolecular interactions, which was also confirmed by the changes in optical properties. In contrast, AuNPs dispersed in lactate and phosphate buffer were stable at therapeutically relevant ionic strength, regardless of their size. Simulation of the local pH distribution near the particle surface also showed a steep pH gradient for particles smaller than 100 nm. This suggests that the healing potential is further enhanced by a more acidic environment at the particle surface, making this strategy a promising approach.
Language	English
Publishing date	2023-06-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2662255-5
ISSN	2079-4991
ISSN	2079-4991
DOI	10.3390/nano13121878
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Article ; Online: Ultrastructural changes in esophageal tissue undergoing stretch tests with possible impact on tissue engineering and long gap esophageal repairs performed under tension.

Biro, Ede / Sommer, Gerhard / Leitinger, Gerd / Abraham, Hajnalka / Kardos, Daniel J / Oberritter, Zsolt / Saxena, Amulya K

Scientific reports

2023 Volume 13, Issue 1, Page(s) 1750

Abstract: Esophageal biomechanical studies are being performed to understand structural changes resulting from stretches during repair of esophageal atresias as well as to obtain biomechanical values for tissue-engineered esophagus. The present study offers ... ...

Abstract	Esophageal biomechanical studies are being performed to understand structural changes resulting from stretches during repair of esophageal atresias as well as to obtain biomechanical values for tissue-engineered esophagus. The present study offers insights into ultrastructural changes after stretching of the ovine esophagus using uniaxial stretch tests. In vitro uniaxial stretching was performed on esophagi (n = 16) obtained from the abattoir within 4-6 h of 1-month-old lambs. Esophagi were divided into 4 groups (4 esophagi/group): control, Group1 (G1), Group2 (G2), Group3 (G3) stretched to 20%, 30% and 40% of their original length respectively. Force and lengthening were measured with 5 cycles performed on every specimen. Transmission electron microscopic (TEM) studies were performed on the 4 groups. During observational TEM study of the control group there were no significant differences in muscle cell structure or extracellular matrix. In all stretched groups varying degrees of alterations were identified. The degree of damage correlated linearly with the increasing level of stretch. Distance between the cells showed significant difference between the groups (control (μ = 0.41 μm, SD = 0.26), G1 (μ = 1.36 μm, SD = 1.21), G2 (μ = 2.8 μm, SD = 1.83), and G3 (μ = 3.01 μm, SD = 2.06). The diameter of the cells (control μ = 19.87 μm, SD = 3.81; G1 μ = 20.38 μm, SD = 4.45; G2 μ = 21.7 μm, SD = 6.58; G3 μ = 24.48 μm, SD = 6.69) and the distance between myofibrils (control μ = 0.23 μm, SD = 0.08; G1 μ = 0.27 μm, SD = 0.08; G2 μ = 0.4 μm, SD = 0.15; G3 μ = 0.61 μm, SD = 0.2) were significantly different as well ( p < 0.05 was considered to be significant). Esophageal stretching > 30% alters the regular intracellular and extracellular structure of the esophageal muscle and leads to disruption of intra- and extracellular bonds. These findings could provide valuable insights into alterations in the microscopic structure of the esophagus in esophageal atresias repaired under tension as well as the basis for mechanical characterization for tissue engineering of the esophagus.
MeSH term(s)	Animals ; Abattoirs ; Esophageal Atresia ; Extracellular Matrix ; Muscle Cells ; Sheep ; Sheep, Domestic ; Tissue Engineering
Language	English
Publishing date	2023-01-31
Publishing country	England
Document type	Journal Article ; Observational Study ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-28894-5
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Article ; Online: A multi-step machine learning approach for accelerating QbD-based process development of protein spray drying.

Fiedler, Daniela / Fink, Elisabeth / Aigner, Isabella / Leitinger, Gerd / Keller, Walter / Roblegg, Eva / Khinast, Johannes G

International journal of pharmaceutics

2023 Volume 642, Page(s) 123133

Abstract: This study proposes a new material-efficient multi-step machine learning (ML) approach for the development of a design space (DS) for spray drying proteins. Typically, a DS is developed by performing a design of experiments (DoE) with the spray dryer and ...

Abstract	This study proposes a new material-efficient multi-step machine learning (ML) approach for the development of a design space (DS) for spray drying proteins. Typically, a DS is developed by performing a design of experiments (DoE) with the spray dryer and the protein of interest, followed by deriving the DoE models via multi-variate regression. This approach was followed as a benchmark to the ML approach. The more complex the process and required accuracy of the final model is, the more experiments are necessary. However, most biologics are expensive and thus experiments should be kept to a minimum. Therefore, the suitability of using a surrogate material and ML for the development of a DS was investigated. To this end, a DoE was performed with the surrogate and the data used for training the ML approach. The ML and DoE model predictions were compared to measurements of three protein-based validation runs. The suitability of using lactose as surrogate was investigated and advantages of the proposed approach were demonstrated. Limitations were identified at protein concentrations >35 mg/ml and particle sizes of x
MeSH term(s)	Spray Drying ; Particle Size
Language	English
Publishing date	2023-06-12
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	428962-6
ISSN	1873-3476 ; 0378-5173
ISSN (online)	1873-3476
ISSN	0378-5173
DOI	10.1016/j.ijpharm.2023.123133
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Article: Investigations to Evaluate Gastric Mucoadhesion of an Organic Product to Ameliorate Gastritis.

Winter, Christina / Hartl, Sonja / Kolb, Dagmar / Leitinger, Gerd / Roblegg, Eva

Pharmaceutics

2020 Volume 12, Issue 4

Abstract: Gastritis is an inflammatory disease leading to abdominal pain, nausea, and diarrhea. While therapy depends on etiology, adhesive agents protecting the gastric tissue represent a promising treatment option. ... ...

Abstract	Gastritis is an inflammatory disease leading to abdominal pain, nausea, and diarrhea. While therapy depends on etiology, adhesive agents protecting the gastric tissue represent a promising treatment option. Caricol
Language	English
Publishing date	2020-04-07
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics12040331
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Article ; Online: Investigation of Changes in Saliva in Radiotherapy-Induced Head Neck Cancer Patients.

Winter, Christina / Keimel, Roman / Gugatschka, Markus / Kolb, Dagmar / Leitinger, Gerd / Roblegg, Eva

International journal of environmental research and public health

2021 Volume 18, Issue 4

Abstract: The intact function of the salivary glands is of utmost importance for oral health. During radiotherapy in patients with head and neck tumors, the salivary glands can be damaged, causing the composition of saliva to change. This leads to xerostomia, ... ...

Abstract	The intact function of the salivary glands is of utmost importance for oral health. During radiotherapy in patients with head and neck tumors, the salivary glands can be damaged, causing the composition of saliva to change. This leads to xerostomia, which is a primary contributor to oral mucositis. Medications used for protective or palliative treatment often show poor efficacy as radiation-induced changes in the physico-chemical properties of saliva are not well understood. To improve treatment options, this study aimed to carefully examine unstimulated whole saliva of patients receiving radiation therapy and compare it with healthy unstimulated whole saliva. To this end, the pH, osmolality, electrical conductivity, buffer capacity, the whole protein and mucin concentrations, and the viscoelastic and adhesive properties were investigated. Moreover, hyaluronic acid was examined as a potential candidate for a saliva replacement fluid. The results showed that the pH of radiation-induced saliva shifted from neutral to acidic, the osmolality increased and the viscoelastic properties changed due to a disruption of the mucin network and a change in water secretion from the salivary glands. By adopting an aqueous 0.25% hyaluronic acid formulation regarding the lost properties, similar adhesion characteristics as in healthy, unstimulated saliva could be achieved.
MeSH term(s)	Head and Neck Neoplasms/radiotherapy ; Humans ; Oral Health ; Saliva ; Xerostomia/etiology
Language	English
Publishing date	2021-02-09
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1660-4601
ISSN (online)	1660-4601
DOI	10.3390/ijerph18041629
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