Microtome Sections Research Articles

Microanatomy sectioning and phytochemicals overview in selected Oleaceae species

This study provides in-depth observations into the microanatomical structure of leaves and stem of eight selected Oleaceae species revealing comprehensive tissue organization using microtome sectioning and peeling leaf epidermis techniques. The phytochemicals profile overview of selected olive species extracted from leaf and stem of Oleaceae species contributes to identifying bioactive compounds with potential therapeutic and medicinal applications. An overview of histochemistry of bioactive metabolites revealed in Oleacee species shows the presence of phenolic compounds (oleuropein and hydroxytyrosol), localized in the leaves and stems, contributing to their antioxidant properties. The present study represent foliar anatomical traits; epidermal cells (ECs) polygonal shape and straight anticlinal walls, paracytic stomata and simple unicellular trichomes. The maximum ECs length (30 μm) was noted in Olea europaea and minimum (20 μm) in Jasminum fluminense was measured. Oleaceae species microtome stem sectioning examined revealed cortex type (parenchyma and collenchyma), vascular bundles (VBs) arrangements (radial, scattered and bundled), xylem vessels (scalariform, annular, spiral and reticulate), phloem fiber arrangements (banded and scattered), and uniseriate, biseriat and multiseriate medullary rays. Stem transverse section with maximum length (1300 μm) was observed in Olea europaea and the lowest (1100 μm) in Jasminum grandiflorum L. subsp. floribundum. This taxonomic histological method links the relationship between anatomy and chemistry, lead to identification of bioactive chemical compounds diversity among Oleaceae species. This anatomical methods used to enhance and consider the plant micromorphological structure-function relationships adapted to diverse environmental conditions.

Data retrieval from archival renal biopsies using nonlinear microscopy.

Thorough examination of renal biopsies may improve understanding of renal disease. Imaging of renal biopsies with fluorescence nonlinear microscopy (NLM) and optical clearing enables three-dimensional (3D) visualization of pathology without microtome sectioning. Archival renal paraffin blocks from 12 patients were deparaffinized and stained with Hoechst and Eosin for fluorescent nuclear and cytoplasmic/stromal contrast, then optically cleared using benzyl alcohol benzyl benzoate (BABB). NLM images of entire biopsy fragments (thickness range 88-660 μm) were acquired using NLM with fluorescent signals mapped to an H&E color scale. Cysts, glomeruli, exudative lesions, and Kimmelstiel-Wilson nodules were segmented in 3D and their volumes, diameters, and percent composition could be obtained. The glomerular count on 3D NLM volumes was high indicating that archival blocks could be a vast tissue resource to enable larger-scale retrospective studies. Rapid optical clearing and NLM imaging enables more thorough biopsy examination and is a promising technique for analysis of archival paraffin blocks.

Rapid Examination of Nonprocessed Renal Cell Carcinoma Using Nonlinear Microscopy.

Histology, the traditional method of examining surgical tissue under a microscope, is a time-consuming process involving the fixation of tissue in formalin, dehydration, embedding in paraffin, and cutting into thin sections for hematoxylin-eosin (H&E) staining. Frozen section analysis is a faster alternative used in surgery to quickly evaluate tissue, but it has limitations, such as the size of the specimens that can be analyzed and difficulties with fatty and bony tissues. To rapidly examine nonprocessed kidney tumors using nonlinear microscopy (NLM), a fluorescence microscopy technique that can rapidly visualize fresh or fixed, rapidly stained, nonprocessed tissue resembling H&E histology. This technology eliminates the need for fixation, embedding, microtome sectioning, or slide preparation. In this study, a total of 190 tissue specimens were collected from 46 patients who underwent partial or radical nephrectomy. Two genitourinary pathologists confirmed that diagnostically important features present in the H&E images could also be identified in the NLM images. The results of this study demonstrated that NLM had a high degree of correspondence with H&E staining for the classical variants of renal cell carcinoma. NLM offers several clinical benefits, such as facilitating rapid renal cell carcinoma diagnosis, assessment of targeted kidney biopsies for both tumor and medical kidney diseases, and collection of fresh renal cell carcinoma tissue for molecular studies.

Cabbage (Brassica oleracea var. capitata) Development in Time: How Differential Parenchyma Tissue Growth Affects Leafy Head Formation.

This study aims to categorize the morphological changes during cabbage (B. oleracea ssp. capitata) development, seedling, rosette, folding, and heading, and to elucidate the cellular mechanisms of the leaf curvature, essential for the formation of the leafy head. We followed the growth of two cabbage cultivars with distinct head shapes (round and pointed) and one non-heading collard cultivar; we phenotyped the size and volume of the whole plant as well as the size, shape, and curvature of the leaves during growth. By integrating these phenotypic data, we determined the four vegetative stages for both cabbages. The histological phenotypes of microtome sections from five distinct leaf positions of the rosette, folding, and heading leaves at two timepoints during leaf growth were quantified and revealed variations in cellular parameters among leaf types, between leaf positions, and between the adaxial and abaxial sides. We identified two synergistic cellular mechanisms contributing to the curvature of heading leaves: differential growth across the leaf blade, with increased growth at the leaf's center relative to the margins; and the increased expansion of the spongy parenchyma layer compared to the palisade parenchyma layer, resulting in the direction of the curvature, which is inwards. These two processes together contribute to the typical leafy heads of cabbages.

Anatomical characteristics and resprouting capacity of the underground organs of Bohemian knotweed (Polygonum ×bohemicum)

AbstractThe hybrid Bohemian knotweed [Polygonum ×bohemicum (J. Chrtek & Chrtková) Zika & Jacobson [cuspidatum × sachalinense]; syn.: Reynoutria ×bohemica Chrtek & Chrtková] is part of the worldwide problematic rhizomatous invasive plants that impact (semi-)natural and agricultural systems. In this context, precise knowledge about the morpho-anatomy and resprouting capacity of the underground organs is key information for developing efficient eradication measures. In the present study, we aimed at (1) clarifying existing differences in the morpho-anatomical characteristics of rhizomes and roots, (2) developing an easy-to-apply field identification method for the underground organs, and (3) identifying the main morpho-anatomical features enhancing the rhizomes’ resprouting ability. For this purpose, we collected the underground organs of two wild populations of P. ×bohemicum in Canton Ticino (southern Switzerland) and analyzed the morpho-anatomical differences between rhizomes and roots, using high-resolution microscope images and microtome sections. Collected material was then used for a resprouting capacity test after assessing rhizome characteristics such as weight, total diameter, pith diameter, pith brightness, and pith color. In contrast to roots, rhizomes are characterized by pith tissue in the center and display nodes with peripheral dormant buds that enable them to resprout. Resprouting ability of rhizomes was high (87.1% on average) and depended on the ontogenetic developmental stage of the organs (peak values of 97% for young and clearer-colored organs, 50% for old and dark ones). In conclusion, the smooth pith tissue of rhizomes represents a key discriminating feature between rhizomes and roots, whereas relating existing nodes to the corresponding rhizome pith color allows assessment of the resprouting potential of a knotweed population.

Characterization of transient and progressive pulmonary fibrosis by spatially correlated phase contrast microCT, classical histopathology and atomic force microscopy

Pulmonary fibrosis (PF) is a severe and progressive condition in which the lung becomes scarred over time resulting in pulmonary function impairment. Classical histopathology remains an important tool for micro-structural tissue assessment in the diagnosis of PF. A novel workflow based on spatial correlated propagation-based phase-contrast micro computed tomography (PBI-microCT), atomic force microscopy (AFM) and histopathology was developed and applied to two different preclinical mouse models of PF - the commonly used and well characterized Bleomycin-induced PF and a novel mouse model for progressive PF caused by conditional Nedd4-2 KO. The aim was to integrate structural and mechanical features from hallmarks of fibrotic lung tissue remodeling. PBI-microCT was used to assess structural alteration in whole fixed and paraffin embedded lungs, allowing for identification of fibrotic foci within the 3D context of the entire organ and facilitating targeted microtome sectioning of planes of interest for subsequent histopathology. Subsequently, these sections of interest were subjected to AFM to assess changes in the local tissue stiffness of previously identified structures of interest. 3D whole organ analysis showed clear morphological differences in 3D tissue porosity between transient and progressive PF and control lungs. By integrating the results obtained from targeted AFM analysis, it was possible to discriminate between the Bleomycin model and the novel conditional Nedd4-2 KO model using agglomerative cluster analysis. As our workflow for 3D spatial correlation of PBI, targeted histopathology and subsequent AFM is tailored around the standard procedure of formalin-fixed paraffin-embedded (FFPE) tissue specimens, it may be a powerful tool for the comprehensive tissue assessment beyond the scope of PF and preclinical research.

Evaluation of the Susceptibility of Lotus Seeds (Nelumbo nucifera Gaertn.) to Aspergillus flavus Infection and Aflatoxin Contamination.

The seeds of lotus (Nelumbo nucifera Gaertn.) have been used as significant medicinal and nutritional ingredients worldwide. The abundant proteins and polysaccharides in lotus seeds make them susceptible to contamination by aflatoxin (AF), a fungal toxic metabolite. This study was conducted to investigate the susceptibility of lotus seeds at different stages of ripening to AF contamination, as well as the mechanism of the contamination. Seven groups of lotus receptacles with seeds at different ripening stages (A-G, from immature to mature) were used for the experiment. Spores of Aspergillus flavus, an AF producer, were inoculated on the water-gap area of the seeds in each receptacle. Then, each receptacle was covered with a sterilized bag, and its stalk part was soaked in water containing a life-prolonging agent, after which it was kept at room temperature for 14 days. The AF content of each whole inoculated seed from the A-G groups and that of each seed part (pericarp, cotyledon, and embryo) from the D and E groups were determined using high-performance liquid chromatography. Microtome sections were prepared from the samples and observed under a light microscope and scanning electron microscope. The seeds from the A and D groups had higher AF contents than the seeds from the B, C, E, F, and G groups, indicating that the condition of the water-gap area and the development of the embryo and cotyledon parts of the seeds are associated with AF contamination.

Floral structure and development in Nandina domestica and implications for systematic affinities in Berberidaceae and perianth evolution in Ranunculales

Abstract In Berberidaceae, the phylogenetic position and systematic affinities of the monotypic genus Nandina have long been debated and remain unclear. In addition, previous comparative studies have provided conflicting reports on the origin and development of its petals. Using scanning electron microscopy and microtome sections, we studied the structure and development of the flower and perianth in Nandina domestica to re-evaluate its affinities and homology of its petals. Our results confirm that N. domestica essentially shares the same Bauplan (organization) with other Berberidaceae and its petals are neither delayed in development nor much differentiated from sepals. However, we sometimes also observed the initiation of additional primordia that become confluent with the base of stamens of the outer whorl, but do not develop further and are no longer visible in later stages of development. Our study thus not only reconciles previous conflicting interpretations on the origin and development of petals in Nandina, but also confirms that it has a unique suite of vegetative and reproductive features supporting its position as a distinct member of the family. In addition, it suggests that the most recent common ancestors of both Berberidaceae and Ranunculales probably had two types of petaloid organs with distinct origin and development but still sharing a deep common genetic identity.

Spatially Resolved Transcriptomics Reveal Changing Landscape of Survival Molecular Signatures in Age-related Macular Degeneration

We study AMD because it accumulates debris beneath the retinal pigment epithelium (RPE), forming so-called “Drusen” analogous to senile plaques of Alzheimer's, including the accumulation of amyloid-beta peptide. We found a decrease in components of the pathway leading to the formation of the pro-homeostatic mediators, the elovanoids (ELVs), specifically in the rod photoreceptors of retinas from donors with age-related macular degeneration (AMD). Retinas were formalin-fixed paraffin-embedded (FFPE), and 5µm microtome sectioning, H&E staining, 10X Genomics Visium CytAssist, and Illumina Sequencing were followed by analysis with Spaceranger and Seurat bioinformatics. By mapping the transcriptome of the human retina, we found gene expression differences in the normal retinas compared to the AMD retinas, both in the rod and cone areas. Transferrin (TF), APOE, and ABCA4 genes were decreased in AMD. Our data have uncovered gene clusters and transcriptomic signatures in regulatory pathways of cell survival in rods and cones in AMD.

Perbedaan Hasil Pewarnaan Hematoxylin Eosin (HE) Pada Histologi Kulit Mencit (Mus Musculus) Berdasarkan Ketebalan Pemotongan 3 Μm, 6 Μm Dan 9 Μm

Sectioning is a step that must be passed before staining Hematoxylin Eosin (HE). The aim of this study was to determine differences in the results of Hematoxylin Eosin (HE) staining in mice skin histology (Mus musculus) based on the thickness of the microtome sections of 3 μm, 6 μm, and 9 μm. Experimental research, true experimental research design post test only control group design. The research sample was mice skin preparations (Mus musculus). Primary data collection, cutting of skin preparations using a microtome and staining Hematoxylin Eosin (HE). Field readings were carried out at 400x magnification (40x objective). Data was processed using Kruskal Wallis and Man Whitney. The average value of 6 μm nuclear cutting was 1.44, cytoplasm was 1.15 and color uniformity was 3. The results showed an abnormal distribution, the Kruskal Wallis test (p=0.008) there was a difference in the quality of staining in the 3 μm, 6 μm and 9 μm. The Man Whitney test for the 3 μm and 6 μm microtome cutting groups (p=0.412) showed no difference in the group, the 6 μm and 9 μm microtome cutting groups (p=0.004) there were differences in the group, the 3 μm and 9 μm cutting groups ( p=0.004) there was a difference in the cutting group. The conclusion of this study was that 6 μm microtome cutting produced preparations with better staining quality compared to 3 μm and 9 μm microtome cutting.

Comparative foliar anatomical study of six different varieties of Piper betle L.

The present investigation on a few selected cultivars of Piper betle L. was undertaken mainly to understand the anatomy and tissue components of the stem, leaves and petiole. This was accomplished by using basic anatomy techniques such as free hand and microtome sectioning. The varieties selected were: HY1, HY2, JB, KB, LV and SG. Major aspects of the present inquiry were - macro morphology, anatomy and histochemistry. The main focus of the study was on the internal structure of leaves and petioles to understand the role of specialised cells like enlarged hypodermal cells and mucilage cavities in retaining moisture and thereby preserving the shelf life of harvested leaves.

Fiber Optic Probe-Based ATR-FTIR Spectroscopy for Rapid Breast Cancer Detection: A Pilot Study.

Breast cancer diagnosis is crucial for timely treatment and improved outcomes. This paper proposes a novel approach for rapid breast cancer diagnosis using optical fiber probe-based attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy from 750 to 4000 cm-1 . The technique enables direct analysis of tissue samples, eliminating the need for microtome sectioning and staining, thus saving time and resources. By capturing molecular fingerprint information, various machine learning models were used to analyze the spectroscopic data to classify cancerous and non-cancerous tissues accurately. Comparing deparaffinized and paraffinized samples reveals the impact of sample preparation and experimental methods. The study demonstrates a strong correlation between the cancerous nature of a sample and its ATR-FTIR spectrum, suggesting its potential for breast cancer diagnosis (sensitivity of 74.2% and specificity of 78.3%). The proposed approach holds promise for integration into clinical operations, providing a rapid method for preliminary breast cancer diagnosis. This article is protected by copyright. All rights reserved.

A Case of Hepatic Cysticercosis in Wistar Albino Rats: An Unintended Finding from an Animal House

Introduction: Rats are the most commonly used animals in biomedical research as they are mild-tempered and docile, making them easy to handle. They breed earlier, produce more offsprings and share many processes with humans and are appropriate for use to answer many research questions. Scientists have specifically bred many strains or lines of rats for research, the most important being the albino Wistar rat. Despite their prolific uses, they have some disadvantages like the spread of diseases.
 Methodology: Wistar albino rats were used in a study at an animal facility by the pharmacology and oral pathology departments to examine the effects of a herbal cardioprotective medicine on rat liver, kidney, and heart function. Also, the Oral cavities were used to observe various stages of tooth development. After the study, the rats were CO2 inhalation-killed in the lab, and their hearts, livers, and kidneys were dissected for examination using a scalpel, and Boyd Parker blades No.3.
 Results: On gross examination, Few animals had several cysts of varied sizes that were macroscopically evident but unwelcome. As a result, all samples were fixed in 10% neutral buffered formalin overnight. The specimens underwent additional processing, including paraffin wax embedding, microtome sectioning, and haematoxylin and eosin staining. Under a light microscope, the slides were examined for histological alterations.
 Analysis: Liver had multiple apparent solitary and multicystic regions, ranging from 1 to 2.5 mm, upon gross examination. On the right lobe, more of these patches with yellowish colour were seen. The cyst's cestode origin was established by histopathological analysis thanks to an exterior acellular eosinophilic cuticular layer, an underlying subtentacular layer, and outer hooks and suckers containing scolex.
 Discussion: The larval stage of Taenia solium, which is frequently found in developing nations, is the infection that results in cysticercosis. Pigs serve as its intermediate hosts whereas humans are its only naturally occurring definitive host. Man may develop into an intermediate host after ingesting adult tapeworm eggs. A condition known as Cysticercosis was the result of this.
 Conclusions: Our findings imply that using rats home to these larvae can lead to inaccurate results in animal research, particularly those that evaluate liver function. They change how the liver functions, producing unreliable results. To properly assess the results of their investigations, the researchers must be aware of the asymptomatic unprompted lesions.

Anatomical Study for Vegetative Organs of Pelargonium graveolens L.'Hér Cultivated in Iraq

The current investigation concluded an anatomical study of the cultivated species Pelargonium graveolens L. 'Hér belong to the Geraniaceae family and which cultivated as an ornamental plant, fresh specimen were collected from different region in Baghdad ,the study vegetative parts (leaf, stem and petiole) in details ,the upper and lower epidermis of leaf and epidermis of stem also studied, ordinary epidermal cells and stomatal complexes studied in term of shapes and dimensions ,the result showed that stomatal in leaf and stem is anomocytic type, the cross sections of the vegetative parts prepared by microtome and free hand sectioning. The tissues of these parts include the number and the type of layers and thickness of each one , the result showed the presence of druses crystals in parenchyma cells of the petiole, stem and leaf. Vistur type of this organs also studied and it comprises of glandular and non-glandular trichomes which occur in different types and lengths.

Seed anatomy and dormancy class of 14 species from the Andean montane forests of Colombia

AbstractSeed dormancy is a key trait used around the world to help understand the ecological dynamics of plant species, however, in some ecosystems such as the Andean forests of Colombia, the identification of dormancy class remains poorly known. To address this, we described the morphology, anatomy and dormancy class of the seeds and fruits of 14 species using microtome sections, light microscopy and scanning electron microscopy. Five native pioneer species showed Morphophysiological Dormancy, three Physical Dormancy + Physiological Dormancy and two Physical Dormancy, whereas two invasive species showed Physiological Dormancy. Only the species of the genus Cecropia had seeds with PY + PD that was promoted by the anatomical structure of the achenes. Of the analysed species, 85% showed dormancy and followed a pattern like other tropical montane ecosystems in the world. The anatomy of achenes and development of seed dormancy play important roles in the capacity of Cecropia species to contribute to the natural regeneration of Andean ecosystems. Additionally, seed dormancy may promote the high invasiveness of Thunbergia alata and Ulex europaeus in the Andean forests of the Central Cordillera of Colombia.

Comparative anatomy and achene micromorphology assessment of two Cousinia Cass. (Asteraceae) species in view of taxonomy

Comparative anatomy and achene micromorphology assessment of two Cousinia Cass. (Asteraceae) 
 species in view of taxonomy
 Deniz ULUKUŞ 1, Osman TUGAY 2
 ORCID: 0000-0002-9627-5492; 0000-0003-3980-7648
 
 1 Department of Biotechnology, Faculty of Sciences, Selçuk University, Konya, Turkey
 2 Department of Pharmaceutical Botany, Faculty of Pharmacy, Selçuk University, Konya, Turkey 
 
 Abstract
 Cousinia is a member of the monophyletic tribe Cardueae, which is traditionally subdivided into four taxonomic groups; however, the rank and boundaries of these taxa are highly controversial. The purpose of this study was to contribute to the genus taxonomy by determining the comparative anatomical and achene micromorphological features of C. aintabensis Boiss. & Hausskn. and C. birecikensis Hub.-Mor. that are morphologically similar to each other. In anatomical studies, stem, leaf, and midrib features were determined. Paraffin embedding, microtome sectioning, and safranin-fast green staining were used for the samples. The obtained sections were photographed, and the leaf, stem, and midrib characters were measured. The importance of anatomical characters between species was determined by applying Independent sample T-test test to quantitative characters. In addition, box plot and heatmap analyses of the studied species were carried out. Our results showed that the stem epidermis, inner sclerenchyma and phloem layer; For leaves; lower and upper epidermis mesophyll thickness, palisade parenchyma; In midrib; number of vascular bundles, collenchyma, and phloem are important characters that can be used in the differentiation of species. SEM microscope was used for achene micromorphological examinations. C. aintabensis achene surface ornamentation is identified as striate-retipylate. It was observed that C. birecikensis had striate and irregular reticulate-faveolate surface ornamentation.
 
 Keywords: Asteraceae, Anatomy, Cousinia, micromorphology, Türkiye

Rhizome architecture, development and vascularization in the water lily Nymphaea alba.

Water lilies are of particular interest with regard to the evolution of angiosperms. They live in an aquatic environment and have been regarded as links to the monocots by some authors. Vascular bundles are sometimes described as scattered or atactostelar as in monocots. However, this view needs to be clarified as the morphology and vascularization of Nymphaea rhizomes remain to be understood. The rhizome of Nymphaea alba was re-investigated morphologically and histologically. Developmental studies were conducted using scanning electron microscopy. Comprehensive histological analyses, including hand and microtome sections and a variety of specific staining procedures, were conducted to re-evaluate the composition of longitudinal and transverse tissue. The rhizome is covered by parenchymatous nodal cushions each bearing a leaf and several adventitious roots. Internodes are extremely short. The apex is flat and early overtopped by developing leaf primordia and cushions. The phyllotaxis is spiral and passes alternately through vegetative and reproductive phases. Flowers appear in the leaf spiral, and lack a subtending bract and a cushion below the peduncle. The reproductive phase includes two or three flowers which alternate with a single leaf. The rhizome is histologically subdivided into a central core, an aerenchymatic cortex, and a parenchymatic exocortex formed to a great extent by the nodal cushions. The core contains strands of vascular bundles united to a complex vascular plexus. Vascular elements continuously anastomose and change shape and direction. Provascular strands originating from leaf primordia merge with the outer core vascular tissue whereas the flower strands run into the centre of the core. Roots originating from the parenchymatous cushions show the characteristic actinostelic pattern, which changes into a collateral pattern inside the rhizome. Several root traces merge and form one strand leading to the central core. Early cell divisions below the apical meristem dislocate leaf, flower and root primordia and their provascular strands outwards. Consequently, fully developed vascular strands insert horizontally into the vascular plexus at advanced rhizome stages. The absence of bracts and cushions below the flowers, the alternate leaf-flower sequence and the course of the peduncle strand suggest that the rhizome is sympodially instead of monopodially organized. The spiral phyllotaxis extends in this case over several shoot orders, masking the branching pattern. The vascular strands in the central plexus differ considerably from vascular bundles in monocots, confirming the unique vascularization in Nymphaea. Sclerenchymatic bundle sheaths are lacking, and vascular bundles continuously split and anastomose throughout the rhizome. Though vascular bundles in petioles and peduncles of N. alba show similarities with some Alismatales, the vascular system of N. alba in general has little in common with that of monocots.

The Mechanism of Ochratoxin Contamination of Artificially Inoculated Licorice Roots.

Ochratoxin (OT) contamination of medicinal herbs is a serious threat to human health. This study was performed to investigate the mechanism of OT contamination of licorice (Glycyrrhiza sp.) root. Licorice root samples were cut into eight parts, which were placed separately on sucrose-free Czapek Dox agar medium, inoculated with the spores of ochratoxigenic Aspergillus westerdijkiae. After incubation for 10 and 20 days, the OT contents of the samples were determined by high-performance liquid chromatography, and microtome sections prepared from the samples were analyzed by desorption electrospray ionization tandem mass spectrometry, to visualize OT localization. The same sections were further examined by light microscopy and scanning electron microscopy, to investigate the path of fungal mycelial penetration of the inner roots. OT concentrations tended to increase from the upper- to the middle-root parts. OTs were located in cut areas and areas of cork layer damage; they were not present in the undamaged cork layer, indicating that the structure of this layer prevents OT contamination of the licorice root.

Idiopathic calcinosis cutis with unusual histomorphology and negative von Kossa stain: A diagnostic pitfall

Introduction: Calcinosis cutis is characterized by basophilic crystalline or amorphous calcium within the dermis/subcutis. Calcinosis cutis has many different clinical subtypes and associated etiologies that may lead clinicians to consider this diagnosis. Von Kossa and/or Alizarin red special stains may be used by pathologists to highlight deposition of insoluble calcium salts, especially in histologically ambiguous or subtle cases. Case Report: We report the case of a 27-year-old male who presented with an inferior right buttock mass clinically diagnosed as epidermal inclusion cyst(s). Gross pathology revealed a gray-white pasty substance within a cyst-like structure. Histopathologic examination demonstrated pools of amorphous blue-gray material with rare deposits of admixed coarse calcifications surrounded by foreign-body giant cells. No cyst wall was seen in the specimen. Special stains, including von Kossa, were initially negative. Following additional review, it was discovered that surface decalcifying solution had been applied to the paraffin block in the histology lab prior to microtome sectioning. Hypothesizing that this could be the cause of the unusual morphology, the paraffin block was reprocessed and subsequent H and E stained sections displayed characteristic basophilic calcium deposits, which were correspondingly positive by von Kossa stain. Conclusion: The histopathologic diagnosis of calcinosis cutis is apparent by HandE in most cases, though von Kossa and/or Alizarin red special stains can be used to aid the pathologist. Given the unusual histomorphology following surface decalcification and initial lack of von Kossa stain prior to reprocessing, this report serves to make pathologists aware of this potential diagnostic pitfall.

Evaluation of Fluoride Uptake of Two Fluoride Varnishes into and onto the Enamel Surface at Different Temperatures: An In Vitro Study.

Fluoride (F) is an effective anticaries agent and can be delivered through various mediums at different concentrations. The ability to increase the enamel resistance to acid by decreasing solubility through the incorporation of F into the enamel apatite structure is the primary function of these agents. The efficacy of topical F can be determined by measuring the amount of F incorporated in and on human enamel. To compare the F uptake into and onto the enamel surface of two different F varnishes at different temperatures. In this study, 96 teeth were divided equally and randomly (n = 48) into two experimental groups (group I and group II). Each group was further divided into four equal subgroups (n = 12); depending on the temperature (25°, 37°, 50°, and 60°C) they were subjected to Fluor-Protector® 0.7% and Embrace® 5% F varnishes were allocated to experimental groups I and II, respectively, and every sample was individually treated with the assigned F varnish. After the varnish application, two specimens from each subgroup, the group I and group II (n = 16), were mounted for hard tissue microtome sectioning for scanning electron microscope (SEM) analysis. The remaining 80 teeth underwent potassium hydroxide (KOH) soluble and KOH-insoluble F estimation. Group I and group II, both, showed maximum F uptake of 2817.07 ppm and 1626.8 ppm at 37°C temperature and the lowest of 1168.9 ppm and 1068.93 ppm at 50°C, respectively. The intergroup comparison was carried out using an unpaired t-test and the intragroup comparison was done using one-way analysis of variance (ANOVA) univariate analysis.Post hoc Tukey test was performed for pairwise comparison between different temperature groups. In group I (Fluor-Protector®), the difference in F uptake was statistically significant when the temperature was increased from 25° to 37° C (mean difference = -9.90, p = 0.016). In group II (Embrace®), a statistically significant difference in F uptake was observed when the temperature was increased from 25° to 50°C (mean difference = 10.00, p = 0.003) and 25° to 60°C (mean difference = 13.38, p = 0.001), respectively. Fluor-Protector® varnish proved to have better F uptake in comparison to Embrace® varnish on human enamel. Topical F varnishes were most effective at 37°C, which is close to the standard human body temperature. Thus, the application of warm F varnish ensures more uptake of F in and onto the enamel surface for greater protection against dental caries. Vishwakarma AP, Bondarde P, Vishwakarma P, et al. Evaluation of Fluoride Uptake of Two Fluoride Varnishes Into and onto the Enamel Surface at Different Temperatures: An In Vitro Study. Int J Clin Pediatr Dent 2022;15(6):672-679.