Light Microscopy Research Articles

A NSC-34 cell line-derived spheroid model: Potential and challenges for in vitro evaluation of neurodegeneration.

Three-dimensional (3D) spheroid models aim to bridge the gap between traditional two-dimensional (2D) cultures and the complex in vivo tissue environment. These models, created by self-clustering cells to mimic a 3D environment with surrounding extracellular framework, provide a valuable research tool. The NSC-34 cell line, generated by fusing mouse spinal cord motor neurons and neuroblastoma cells, is essential for studying neurodegenerative diseases like amyotrophic lateral sclerosis (ALS), where abnormal protein accumulation, such as TAR-DNA-binding protein 43 (TDP-43), occurs in affected nerve cells. However, NSC-34 behavior in a 3D context remains underexplored, and this study represents the first attempt to create a 3D model to determine its suitability for studying pathology. We generated NSC-34 spheroids using a nonadhesive hydrogel-based template and characterized them for 6 days. Light microscopy revealed that NSC-34 cells in 3D maintained high viability, a distinct round shape, and forming stable membrane connections. Scanning electron microscopy identified multiple tunnel-like structures, while ultrastructural analysis highlighted nuclear bending and mitochondria alterations. Using inducible GFP-TDP-43-expressing NSC-34 spheroids, we explored whether 3D structure affected TDP-43 expression, localization, and aggregation. Spheroids displayed nuclear GFP-TDP-43 expression, albeit at a reduced level compared with 2D cultures and generated both TDP-35 fragments and TDP-43 aggregates. This study sheds light on the distinctive behavior of NSC-34 in 3D culture, suggesting caution in the use of the 3D model for ALS or TDP-43 pathologies. Yet, it underscores the spheroids' potential for investigating fundamental cellular mechanisms, cell adaptation in a 3D context, future bioreactor applications, and drug penetration studies. RESEARCH HIGHLIGHTS: 3D spheroid generation: NSC-34 spheroids, developed using a hydrogel-based template, showed high viability and distinct shapes for 6 days. Structural features: advanced microscopy identified tunnel-like structures and nuclear and mitochondrial changes in the spheroids. Protein dynamics: the study observed how 3D structures impact TDP-43 behavior, with altered expression but similar aggregation patterns to 2D cultures. Research implications: this study reveals the unique behavior of NSC-34 in 3D culture, suggests a careful approach to use this model for ALS or TDP-43 pathologies, and highlights its potential in cellular mechanism research and drug testing applications.

Discovery of pararealgar and semi-amorphous pararealgar in Rembrandt's The Night Watch: analytical study and historical contextualization

This article reports on the discovery of pararealgar and semi-amorphous pararealgar in Rembrandt's masterpiece The Night Watch. A large-scale research project named Operation Night Watch was started in 2019. A variety of non-invasive analytical imaging techniques, together with paint sample research, has provided new information about Rembrandt's pigments, materials, and techniques as well as the current condition of the painting. Macroscopic X-ray fluorescence, macroscopic X-ray powder diffraction and reflectance imaging spectroscopy identified the presence of arsenic sulfide pigments and degradation products of these pigments in the doublet sleeves and embroidered buff coat worn by Lieutenant Willem van Ruytenburch (central figure to the right of Captain Frans Banninck Cocq). Examination by light microscopy of two paint samples taken from this area shows a mixture of large sharp-edged tabular yellow and orange to red pigment particles, and scanning electron microscopy-energy dispersive X-ray analysis identified these particles as containing arsenic and sulfur. Using micro-Raman spectroscopy, the yellow particles were identified as pararealgar, and the orange to red particles as semi-amorphous pararealgar. Synchrotron-based X-ray diffraction allowed visualization of the presence of multiple degradation products associated with arsenic sulfides throughout the paint layer. The discovery of pararealgar and semi-amorphous pararealgar is a new addition to Rembrandt's pigment palette. To contextualize our findings and to hypothesize why, how, and where Rembrandt obtained the pigments, we studied related historical sources. A comprehensive review of historical sources gives insight into the types of artificial arsenic sulfides that were available and suggests that a broader range of arsenic pigments could have been available in Amsterdam in the seventeenth century than previously thought. This is supported by the use of a very similar mixture of pigments by Willem Kalf (1619–1693), a contemporary artist based in Amsterdam. Together with the condition of the particles in the paint cross sections, this brings us to the conclusion that Rembrandt intentionally used pararealgar and semi-amorphous pararealgar, together with lead–tin yellow and vermilion, to create an orange paint.

Outbreak of Feline Sporotrichosis with Zoonotic Potential in the Seventh Health District of Maceió-AL

Sporotrichosis is a mycosis with zoonotic potential caused by species of Sporothrix. Once thought rare in northeastern Brazil, the disease has recently been spreading, leading to an emergency health issue. In this paper, we describe an outbreak of feline sporotrichosis in the Seventh Health District of Maceió-AL. We collected samples from 23 domiciled and non-domiciled felines without regard for age, breed, sex, and neutering state. Skin samples were analyzed cytologically under a light microscope and seeded onto Sabouraud dextrose agar at 25 °C for from 15 to 30 days. Fifteen of the twenty-three cats with suspected skin lesions were positive for Sporothrix spp. on either cytological or microbiological evaluation. Most of the infected cats were male, young adults, non-neutered, with free access to external areas, and living in environments with poor sanitation, a high population density, and an accumulation of garbage and organic matter. Three owners were bitten or scratched by infected cats and subsequently developed suspicious cutaneous lesions suggestive of sporotrichosis. The epidemiological features of feline sporotrichosis in the outbreaks of Maceió seemed to share similarities with the data obtained from outbreaks in current hyperendemic areas. Identifying geographical sites of infection and providing compulsory notification of the disease is essential for avoiding an epidemic in Alagoas.

Modification of the Structure and Mechanical Properties of Cu-30%Zn alloy with Cerium and Silicon

This study investigates how the introduction of cerium and silicon influences the structure and mechanical characteristics of Cu-30%Zn alloy. The examined properties included tensile strength, hardness, and impact resistance. The tensile strength of the developed alloys was determined using an automated JPL tensile strength tester (Model: 130812) with a capacity of 100 kilonewtons, following the ASTM D638 standard. The impact strength was assessed by the ASTM D256 standard using a pendulum impact testing machine (Model: U1820). Additionally, hardness testing was conducted using a Brinell tester. The Brinell hardness test was performed with a portable dynamic hardness tester (Model: DHT-6) by the British Standard (BS EN ISO 6505-1:2014). Specimens were prepared by incorporating cerium and silicon into the Cu-30%Zn alloy at concentrations of 0.1wt%, 0.3wt%, 0.5wt%, 1.0wt%, 3.0wt% 5.0wt% ranging from 0.1% to 5.0%, with increments of 0.2% for micro addition and 2.0% for macro addition. Microstructural analysis was performed using an L2003A reflected light metallurgical microscope. Scanning Electron Microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) were also utilized to characterize the cast specimens. The findings revealed a proportional increase in tensile strength, impact strength, and hardness strength with the rise in dopant concentrations up to 3.0wt% for cerium and silicon. Also, the analysis uncovered the presence of. primary α-phase, β-phase (intermetallic phases), and fine stable phase. These phase modifications contributed to the enhancement of mechanical properties. Cu-30wt%Zn enriched with 3.0% cerium and silicon demonstrated heightened tensile strength, impact strength, and hardness, making it a recommended choice for engineering applications.

Impact of clay minerals on reservoir sandstone properties: comparative study in Colombian eastern cordillera and middle Magdalena valley basins

The aim of this study is to examine how mineralogy influences the petrophysical properties, particularly porosity and permeability, of potential sandstone reservoirs in Colombia. It seeks to comprehensively understand how the presence of clay minerals impacts the overall quality of hydrocarbon reservoirs in the country. Samples of sandstones from reservoirs at various outcrops in the Eastern Cordillera and Middle Magdalena Valley basins were collected. Detailed analysis of mineralogy and petrographic characteristics of the samples was conducted through various analytical techniques such as transmitted light microscopy, X-ray diffraction, and scanning electron microscopy. Porosity and permeability were measured using automated permeametry and porosimetry equipment. The predominant composition of the analyzed reservoir rocks comprises quartz (45-50%), feldspar (35-40%), and clays (10-20%). These rocks were categorized into two distinct groups based on their permeability (K) and porosity (Φ), ranging from 0.009 to 29.220 mD and 1.88 to 20.75%, respectively. The presence of illite correlated with a reduction in both porosity and permeability, highlighting its negative impact on reservoir quality. Conversely, an elevated concentration of kaolinite was associated with favorable porosity and permeability. Samples with feldspar sericitization demonstrated inferior hydrocarbon storage quality. This study provides a deeper understanding of how mineralogy affects the petrophysical properties of sandstone reservoirs in Colombia. These findings are crucial for guiding exploration and production strategies in the Colombian oil industry, especially in challenging geological environments like those studied.

Prototyping in Polymethylpentene to Enable Oxygen-Permeable On-a-Chip Cell Culture and Organ-on-a-Chip Devices Suitable for Microscopy

With the rapid development and commercial interest in the organ-on-a-chip (OoC) field, there is a need for materials addressing key experimental demands and enabling both prototyping and large-scale production. Here, we utilized the gas-permeable, thermoplastic material polymethylpentene (PMP). Three methods were tested to prototype transparent PMP films suitable for transmission light microscopy: hot-press molding, extrusion, and polishing of a commercial, hazy extruded film. The transparent films (thickness 20, 125, 133, 356, and 653 µm) were assembled as the cell-adhering layer in sealed culture chamber devices, to assess resulting oxygen concentration after 4 days of A549 cell culture (cancerous lung epithelial cells). Oxygen concentrations stabilized between 15.6% and 11.6%, where the thicker the film, the lower the oxygen concentration. Cell adherence, proliferation, and viability were comparable to glass for all PMP films (coated with poly-L-lysine), and transparency was adequate for transmission light microscopy of adherent cells. Hot-press molding was concluded as the preferred film prototyping method, due to excellent and reproducible film transparency, the possibility to easily vary film thickness, and the equipment being commonly available. The molecular orientation in the PMP films was characterized by IR dichroism. As expected, the extruded films showed clear orientation, but a novel result was that hot-press molding may also induce some orientation. It has been reported that orientation affects the permeability, but with the films in this study, we conclude that the orientation is not a critical factor. With the obtained results, we find it likely that OoC models with relevant in vivo oxygen concentrations may be facilitated by PMP. Combined with established large-scale production methods for thermoplastics, we foresee a useful role for PMP within the OoC field.

Cytotoxicity and Genotoxicity Effects of a Magnetic Zeolite Composite in Daphnia magna (Straus, 1820)

Zeolite type 5A combined with the magnetic properties of maghemite nanoparticles facilitate the rapid absorption of heavy metals, which makes them an interesting proposal for the remediation of water contaminated with lead and arsenic. However, the physicochemical analysis related to concentration and size for the use of this magnetic zeolite composite (MZ0) in water bodies and the possible toxicological effects on aquatic fauna has not yet been carried out. The main objective of the research work is to determine lethal concentrations that cause damage to Daphnia magna based on LC50 tests, morphology, reproductive rate, and quantification of the expression of three genes closely involved in the morphological development of vital structures (Glass, NinaE, Pph13). To achieve this objective, populations of neonates and young individuals were used, and results showed that the LC50 for neonates was 11,314 mg L−1, while for young individuals, it was 0.0310 mg L−1. Damage to morphological development was evidenced by a decrease in eye size in neonates, an increase in eye size in young individuals, variations in the size of the caudal spine for both age groups, and slight increases in the heart size, body, and antenna for both age groups. The reproductive rate of neonates was not affected by the lower concentrations of MZ0, while in young individuals, the reproductive rate decreased by more than 50% from the minimum exposure concentration of MZ0. And for both ages, Glass gene expression levels decreased as the MZ0 concentration increased. Also, the MZ0 evidenced its affinity for the exoskeleton of D. magna, which was observed using both light microscopy and electron microscopy. It is concluded that MZ0 did not generate significant damage in the mortality, morphology, reproductive rate, or gene expression in D. magna at lower concentrations, demonstrating the importance of evaluating the possible impacts on different life stages of the cladoceran.

Cytotoxicity and Genotoxicity Effects of a Magnetic Zeolite Composite in Daphnia magna (Straus, 1820)

Zeolite type 5A combined with the magnetic properties of maghemite nanoparticles facilitate the rapid absorption of heavy metals, which makes them an interesting proposal for the remediation of water contaminated with lead and arsenic. However, the physicochemical analysis related to concentration and size for the use of this magnetic zeolite composite (MZ0) in water bodies and the possible toxicological effects on aquatic fauna has not yet been carried out. The main objective of the research work is to determine lethal concentrations that cause damage to Daphnia magna based on LC50 tests, morphology, reproductive rate, and quantification of the expression of three genes closely involved in the morphological development of vital structures (Glass, NinaE, Pph13). To achieve this objective, populations of neonates and young individuals were used, and results showed that the LC50 for neonates was 11,314 mg L−1, while for young individuals, it was 0.0310 mg L−1. Damage to morphological development was evidenced by a decrease in eye size in neonates, an increase in eye size in young individuals, variations in the size of the caudal spine for both age groups, and slight increases in the heart size, body, and antenna for both age groups. The reproductive rate of neonates was not affected by the lower concentrations of MZ0, while in young individuals, the reproductive rate decreased by more than 50% from the minimum exposure concentration of MZ0. And for both ages, Glass gene expression levels decreased as the MZ0 concentration increased. Also, the MZ0 evidenced its affinity for the exoskeleton of D. magna, which was observed using both light microscopy and electron microscopy. It is concluded that MZ0 did not generate significant damage in the mortality, morphology, reproductive rate, or gene expression in D. magna at lower concentrations, demonstrating the importance of evaluating the possible impacts on different life stages of the cladoceran.

Minimal Detectable Bone Fracture Gaps in CT Images and Digital Three-Dimensional (3D) Radii Models.

Knowledge of the minimal detectable bone fracture gap is essential in three-dimensional (3D) models, particularly in pre-operative planning of osteosynthesis to avoid overlooking gaps. In this study, defined incisions and bony displacements ranging from 100 to 400 µm were created in diaphyseal radii in 20 paired forearm specimens and verified with light microscopy. The specimens were scanned utilizing different computed tomography (CT) technologies/scanners, specimen positionings, scan protocols, image segmentations, and processing protocols. Inter- and intra-operator variabilities were reported as coefficient kappa. In CT images, fracture gaps of 100 µm and bone lamellae of 300 µm and 400 µm width were identified at a rate of 80 to 100%, respectively, independent of the investigated settings. In contrast, only 400µm incisions and bony displacements were visible in digital 3D models, with detection rates dependent on CT technology, image segmentation, and post-processing algorithm. 3D bone models based on state-of-the-art CT imaging can reliably visualize clinically relevant bone fracture gap sizes. However, verification of fractures to be surgically addressed should be verified with the original CT image series.

Histomorphometric Study of the Tunics of Ductus Arteriosus in Human Fetal Cadavers Using the ImageJ Software.

The ductus arteriosus (DA) connects the left pulmonary artery with theaorta during fetal life. Although it connects two elastic arteries, histological studies have shown that it is a muscular artery. There are very few studies on the histomorphometry of human fetal cadaveric DA. There are few studies on the changes in the tunics of the DA at various stages of fetal development. The present study aimed to observe the histomorphometric features ofDA and its histological variations according to the gestational age of the fetus. The study sample was DA dissected from 34 fetal cadavers of different gestational ages and stained with standardhematoxylin and eosin staining (H&E). We studied the structure of DA under a light microscope. We used ImageJ software (National Institutes of Health, Bethesda, Maryland) to measure the thickness of all three layers of the DA wall. The thickness of the DA wall was directly proportional to the gestational age of the fetus. In each trimester, we observed distinct histological changes in the tunics. The formation of multiple intimal mounds and the increase in intimal thickness observed during the last trimester are responsible for the closure of the ductus after birth. Elastosis is associated with patent DA. The disappearance of elastosis at a later gestational age ensures the closure of the ductus.

Exogenous Sodium Nitroprusside Affects the Redox System of Wheat Roots Differentially Regulating the Activity of Antioxidant Enzymes under Short-Time Osmotic Stress

Nitric oxide (NO) is a multifunctional signalling molecule involved in the regulation of plant ontogenesis and adaptation to different adverse environmental factors, in particular to osmotic stress. Understanding NO-induced plant protection is important for the improvement of plant stress tolerance and crop productivity under global climate changes. The root system is crucial for plant survival in a changeable environment. Damages that it experiences under water deficit conditions during the initial developmental periods seriously affect the viability of the plants. This work was devoted to the comparative analysis of the pretreatment of wheat seedlings through the root system with NO donor sodium nitroprusside (SNP) for 24 h on various parameters of redox homeostasis under exposure to osmotic stress (PEG 6000, 12%) over 0.5–24 h. The active and exhausted solutions of SNP, termed as (SNP/+NO) and (SNP/−NO), respectively, were used in this work at a concentration of 2 × 10−4 M. Using biochemistry and light microscopy methods, it has been revealed that osmotic stress caused oxidative damages and the disruption of membrane cell structures in wheat roots. PEG exposure increased the production of superoxide (O2•−), hydrogen peroxide (H2O2), malondialdehyde (MDA), and the levels of electrolyte leakage (EL) and lipid peroxidation (LPO). Stress treatment enhanced the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), the excretion of proline, and the rate of cell death and inhibited their division. Pretreatment with (SNP/+NO) decreased PEG-induced root damages by differently regulating the antioxidant enzymes under stress conditions. Thus, (SNP/+NO) pretreatment led to SOD, APX, and CAT inhibition during the first 4 h of stress and stimulated their activity after 24 h of PEG exposure when compared to SNP-untreated or (SNP/−NO)-pretreated and stress-subjected plants. Osmotic stress triggered the intense excretion of proline by roots into the external medium. Pretreatment with (SNP/+NO) in contrast with (SNP/−NO) additionally increased stress-induced proline excretion. Our results indicate that NO is able to mitigate the destructive effects of osmotic stress on the roots of wheat seedlings. However, the mechanisms of NO protective action may be different at certain periods of stress exposure.

Prevalence of Clinical malaria and associated symptoms in pregnant women at Hamusit Health Center, Northwest Ethiopia

BackgroundMalaria is the disease caused by intracellular parasites known as Plasmodium species and is mainly transmitted by blood sucking female Anopheles mosquitoes. During pregnancy, malaria results in severe complications to the mother, the fetus and the newborn. Symptoms of malaria, such as fever, malaise, headache, nausea and vomiting, in pregnant women can be mistakenly attributed solely to pregnancy. In Ethiopia, the prevalence of malaria in asymptomatic pregnant women has been well documented. However, studies indicating the prevalence and clinical presentation of malaria in pregnant women are lacking. Therefore, there is little information on the prevalence of malaria and significantly associated signs and symptoms in pregnant women. The aim of this cross-sectional study was to determine the prevalence of malaria and identify clinical signs and symptoms associated with malaria which suggest presence of malaria in pregnant women at Hamusit Health Center, Northwest Ethiopia. MethodsA health facility-based cross-sectional study was conducted on 231 malaria symptomatic pregnant women from April to June 2023. A convenience sampling technique was employed. The socio demographic and clinical data of the study participants was collected through face-to-face interview using questionnaire. Thick and thin blood films were prepared from capillary blood and stained with 10% Giemsa. The stained blood smear was washed with clean water, air dried and examined under a light microscope. The Statistical Package for Social Sciences software version 20 (SPSS 20) was used to analyses data. Logistic regression was used to assess signs and symptoms associated with malaria. An adjusted odds ratio with a 95% confidence interval was calculated, and a P value < 0.05 was considered statistically significant. ResultsThe overall prevalence of malaria among symptomatic pregnant women in the study area was 22.9% (53/231) (95% CI: 17.3-29%). The most prevalent species was P. falciparum, with a frequency of 14.3% (33/231) (95% CI: 10% -18.6%), followed by P. vivax, 5.2% (12/231) (95% CI: (2.6% – 8.2%). The remaining 3.5% (8/231) (95% CI: 1.8% – 6.7%) were mixed infections of P. falciparum & P. vivax. Primigravidae (62.3%) and first trimester pregnancies (52.8%) were more affected. Malaria signs and symptoms mainly, fever [(P=0.002, AOR (95%CI); 5.1(1.84, 14.30)], joint pain [(P=0.001, AOR (95%CI); 7.8(2.24, 27.32)], vomiting [(P=0.007, AOR (95%CI); 2.9(1.34, 6.43)], malaise [(P=0.005, AOR (95%CI); 3.6(1.48, 8.67)] and fatigue [(P=0.0039, AOR (95%CI); 2.1(1.04, 4.37)], were significantly associated with malaria infection in pregnant women. ConclusionsMalaria positivity in pregnant women with fever, joint pain, vomiting, malaise and fatigue is considerably high in the study area. These signs and symptoms in pregnant women are strong indicators of malaria infection.

Mitochondrial Structure, Dynamics, and Physiology: Light Microscopy to Disentangle the Network.

Mitochondria serve as energetic and signaling hubs of the cell: This function results from the complex interplay between their structure, function, dynamics, interactions, and molecular organization. The ability to observe and quantify these properties often represents the puzzle piece critical for deciphering the mechanisms behind mitochondrial function and dysfunction. Fluorescence microscopy addresses this critical need and has become increasingly powerful with the advent of superresolution methods and context-sensitive fluorescent probes. In this review, we delve into advanced light microscopy methods and analyses for studying mitochondrial ultrastructure, dynamics, and physiology, and highlight notable discoveries they enabled.

Reproducibility of c-Met Immunohistochemical Scoring (Clone SP44) for Non-Small Cell Lung Cancer Using Conventional Light Microscopy and Whole Slide Imaging.

Emerging therapies for non-small cell lung cancer targeting c-Met overexpression have recently demonstrated promising results. However, the evaluation of c-Met expression can be challenging. We aimed to study the inter and intraobserver reproducibility of c-Met expression evaluation. One hundred ten cases with non-small cell lung cancer (40 biopsies and 70 surgical specimens) were retrospectively selected in a single laboratory (LPCE) and evaluated for c-Met expression. Six pathologists (4 seniors and 2 juniors) evaluated the H-score and made a 3-tier classification of c-Met expression for all cases, using conventional light microscopy (CLM) and whole slide imaging (WSI). The interobserver reproducibility with CLM gave global Cohen Kappa coefficients (ƙ) ranging from 0.581 (95% CI: 0.364-0.771) to 0.763 (95% CI: 0.58-0.92) using the c-Met 3-tier classification and H-score, respectively. ƙ was higher for senior pathologists and biopsy samples. The interobserver reproducibility with WSI gave a global ƙ ranging from 0.543 (95% CI: 0.33-0.724) to 0.905 (95% CI: 0.618-1) using the c-Met H-score and 2-tier classification (≥25% 3+), respectively. ƙ for intraobserver reproducibility between CLM and WSI ranged from 0.713 to 0.898 for the c-Met H-score and from 0.600 to 0.779 for the c-Met 3-tier classification. We demonstrated a moderate to excellent interobserver agreement for c-Met expression with a substantial to excellent intraobserver agreement between CLM and WSI, thereby supporting the development of digital pathology. However, some factors (scoring method, type of tissue samples, and expertise level) affect reproducibility. Our findings highlight the importance of establishing a consensus definition and providing further training, particularly for inexperienced pathologists, for c-Met immunohistochemistry assessment in clinical practice.

Reproducibility of c-Met Immunohistochemical Scoring (Clone SP44) for Non-Small Cell Lung Cancer Using Conventional Light Microscopy and Whole Slide Imaging.

Emerging therapies for non-small cell lung cancer targeting c-Met overexpression have recently demonstrated promising results. However, the evaluation of c-Met expression can be challenging. We aimed to study the inter and intraobserver reproducibility of c-Met expression evaluation. One hundred ten cases with non-small cell lung cancer (40 biopsies and 70 surgical specimens) were retrospectively selected in a single laboratory (LPCE) and evaluated for c-Met expression. Six pathologists (4 seniors and 2 juniors) evaluated the H-score and made a 3-tier classification of c-Met expression for all cases, using conventional light microscopy (CLM) and whole slide imaging (WSI). The interobserver reproducibility with CLM gave global Cohen Kappa coefficients (ƙ) ranging from 0.581 (95% CI: 0.364-0.771) to 0.763 (95% CI: 0.58-0.92) using the c-Met 3-tier classification and H-score, respectively. ƙ was higher for senior pathologists and biopsy samples. The interobserver reproducibility with WSI gave a global ƙ ranging from 0.543 (95% CI: 0.33-0.724) to 0.905 (95% CI: 0.618-1) using the c-Met H-score and 2-tier classification (≥25% 3+), respectively. ƙ for intraobserver reproducibility between CLM and WSI ranged from 0.713 to 0.898 for the c-Met H-score and from 0.600 to 0.779 for the c-Met 3-tier classification. We demonstrated a moderate to excellent interobserver agreement for c-Met expression with a substantial to excellent intraobserver agreement between CLM and WSI, thereby supporting the development of digital pathology. However, some factors (scoring method, type of tissue samples, and expertise level) affect reproducibility. Our findings highlight the importance of establishing a consensus definition and providing further training, particularly for inexperienced pathologists, for c-Met immunohistochemistry assessment in clinical practice.

Generative Modelling of Cortical Receptor Distributions from Cytoarchitectonic Images in the Macaque Brain.

Neurotransmitter receptor densities are relevant for understanding the molecular architecture of brain regions. Quantitative in vitro receptor autoradiography, has been introduced to map neurotransmitter receptor distributions of brain areas. However, it is very time and cost-intensive, which makes it challenging to obtain whole-brain distributions. At the same time, high-throughput light microscopy and 3D reconstructions have enabled high-resolution brain maps capturing measures of cell density across the whole human brain. Aiming to bridge gaps in receptor measurements for building detailed whole-brain atlases, we study the feasibility of predicting realistic neurotransmitter density distributions from cell-body stainings. Specifically, we utilize conditional Generative Adversarial Networks (cGANs) to predict the density distributions of the M2 receptor of acetylcholine and the kainate receptor for glutamate in the macaque monkey's primary visual (V1) and motor cortex (M1), based on light microscopic scans of cell-body stained sections. Our model is trained on corresponding patches from aligned consecutive sections that display cell-body and receptor distributions, ensuring a mapping between the two modalities. Evaluations of our cGANs, both qualitative and quantitative, show their capability to predict receptor densities from cell-body stained sections while maintaining cortical features such as laminar thickness and curvature. Our work underscores the feasibility of cross-modality image translation problems to address data gaps in multi-modal brain atlases.

Addressing Glass Contamination in Radiology: What Can We Do to Minimize Its Impact?

Abstract Introduction There is a risk of glass contamination with the use of single-dose glass ampoules. Complications of injection include infection and granuloma formation and this is widely described in anesthetic literature. To date, there is no data on the effect of different ampoule opening methods on the degree of glass contamination. Purpose This article explores different ampoule opening methods and determines if any method is superior to the others with respect to glass contamination frequency. This article also increases awareness of glass contamination and its potential complications in the radiology community. Methods A controlled trial was undertaken with 15 glass ampoules filled with normal saline, divided into three groups. The ampoules in each of the group were opened via each method: freehand, ampoule breaker, and ampoule opener. The solution was aspirated with an 18-gauge drawing-up needle, which was centrifuged and decantered to be placed onto slides and inspected under light microscopy to assess the glass contaminants. Results Between each cohort, the freehand opening provided the least number of glass particles with 42, followed by the ampoule breaker and snapper. The greatest size of glass contamination was seen from the ampoule snapper at 300 μm, while the lowest average particle size was seen from the ampoule breaker. Conclusion The study confirmed presence of glass contamination in all three methods. Freehand opening minimized the number of particulates, while the ampoule breaker minimized the average particulate size. The ampoule snapper produced larger glass particulates in the trials and was deemed the least effective method.

Addressing Glass Contamination in Radiology: What Can We Do to Minimize Its Impact?

Abstract Introduction There is a risk of glass contamination with the use of single-dose glass ampoules. Complications of injection include infection and granuloma formation and this is widely described in anesthetic literature. To date, there is no data on the effect of different ampoule opening methods on the degree of glass contamination. Purpose This article explores different ampoule opening methods and determines if any method is superior to the others with respect to glass contamination frequency. This article also increases awareness of glass contamination and its potential complications in the radiology community. Methods A controlled trial was undertaken with 15 glass ampoules filled with normal saline, divided into three groups. The ampoules in each of the group were opened via each method: freehand, ampoule breaker, and ampoule opener. The solution was aspirated with an 18-gauge drawing-up needle, which was centrifuged and decantered to be placed onto slides and inspected under light microscopy to assess the glass contaminants. Results Between each cohort, the freehand opening provided the least number of glass particles with 42, followed by the ampoule breaker and snapper. The greatest size of glass contamination was seen from the ampoule snapper at 300 μm, while the lowest average particle size was seen from the ampoule breaker. Conclusion The study confirmed presence of glass contamination in all three methods. Freehand opening minimized the number of particulates, while the ampoule breaker minimized the average particulate size. The ampoule snapper produced larger glass particulates in the trials and was deemed the least effective method.

Autophagy in the corpus luteum correlates with tissue growth in pregnant rats.

The developmental activation of the corpus luteum (CL) structurally and functionally is critical for the temporally regulated establishment, maintenance, and termination of pregnancy in rats. In this study, we have investigated the possible involvement of autophagy in the regulation of the CL during pregnancy in rats. The expression ratio of microtubule-associated protein light chain 3 (LC3)-II/-I, a widely used indicator of autophagic activity, in the CL remained relatively stable until day 15 of pregnancy. Subsequently, it progressively increased until day 21, and then declined until day 3 postpartum. This fluctuation was closely associated with the tissue weight of the CL rather than progesterone (P4) production activity. Light and electron microscopy revealed the presence of immunoreactive LC3 aggregates and irregularly shaped autolysosome-like microstructures in the cytoplasm of luteal cells during late pregnancy. Notably, a bolus intrabursal injection of the autophagy inhibitor bafilomycin A1 on day 15 of pregnancy resulted in a significant reduction in luteal cell size and disrupted the normal alteration of circulating P4 levels. Consequently, treatment with this inhibitor increased the likelihood of the varied timing (both advanced and delayed) of delivery and led to reduced body weight in neonates when compared with the vehicle-treated control group. Our findings suggest that autophagy in the rat CL contributes to luteal tissue growth, influences P4 production, and thereby fine-tunes the regulation of gestation length in rats.

Molecular monitoring of Dinophysis species assemblage in mussel farms in the Northwestern Adriatic Sea

Several Dinophysis species can produce potent lipophilic toxins that pose a risk to human health when contaminated seafood is consumed, especially filter-feeding bivalve mussels. In the mussel farms of the Northwestern Adriatic Sea, seawater and seafood are regularly monitored for the presence of Dinophysis species and their associated toxins, but the current methodological approaches, such as light microscopy determinations, require a long time to make results available to local authorities. A molecular qPCR-based assay can be used to quantify various toxic Dinophysis species in a shorter timeframe. However, this approach is not currently employed in official testing activities. In this study, field samples were collected monthly or bi-weekly over one year from various mussel farms along the Northwestern Adriatic coast. The abundance of Dinophysis species in the seawater was determined using both traditional microscopy and qPCR assays. In addition, the concentration of lipophilic toxins for DSP in mussel flesh was quantified using LC-MS/MS focusing on the okadaic acid group. Dinophysis spp. site-specific single cells were isolated and analysed by qPCR yielding a mean rDNA copy number per cell of 1.21 × 104 ± 1.81 × 103. The qPCR assay gave an efficiency of 98 % and detected up to 10 copies of the rDNA target gene. The qPCR and light microscopy determinations in environmental samples showed a significant positive correlation (Spearman rs = 0.57, p-value < 0.001) with a ratio of 2.24 between the two quantification methods, indicating that light microscopy estimates were generally 44.6 % lower than those obtained by the qPCR assay. The qPCR approach showed several advantages such as rapidity, sensitivity and efficiency over conventional microscopy analysis, showing its potential future role in phytoplankton monitoring under the Official Controls Regulations for shellfish.