Visibility of esophageal squamous cell carcinoma under iodine staining on texture and color enhancement imaging.

Objective: Iodine staining on white light imaging (WLI) is the gold standard for detecting and demarcating esophageal squamous cell carcinoma (ESCC). We examined the effects of texture and color enhancement imaging (TXI) on improving the endoscopic visibility of ESCC under iodine staining. Methods: Twenty ESCC lesions that underwent endoscopic submucosal dissection were retrospectively included. The color difference between ESCC and the surrounding mucosa (ΔEe) on WLI, TXI, and narrow-band imaging was assessed, and ΔEe under 1% iodine staining on WLI and TXI. Furthermore, the visibility grade determined by endoscopists was evaluated on each imaging. Result: The median ΔEe was greater on TXI than on WLI (14.53 vs. 10.71, respectively; p < 0.005). Moreover, the median ΔEe on TXI under iodine staining was greater than the median ΔEe on TXI and narrow-band imaging (39.20 vs. 14.53 vs. 16.42, respectively; p < 0.005 for both). A positive correlation in ΔEe under iodine staining was found between TXI and WLI (correlation coefficient = 0.61, p < 0.01). Moreover, ΔEe under iodine staining on TXI in each lesion was greater than the corresponding ΔEe on WLI. The visibility grade assessed by endoscopists on TXI was also significantly greater than that on WLI under iodine staining (p < 0.01). Conclusions: The visibility of ESCC after iodine staining was greater on TXI than on WLI.

Efficacy of Keratin AE1/AE3 Staining in Evaluating Resection Margins of a Urachal Remnant and Improving Surgical Precision and Outcomes: A Case Report.

A urachal remnant is a rare condition characterized by the persistence of the urachus beyond birth, often presenting with symptoms such as umbilical effusion, periomphalitis, and abdominal pain. Surgical resection is the cornerstone of treatment, but ensuring complete removal of urachal epithelium at the resection margin remains a challenge. This case report focuses on evaluating resection margins of urachal remnants and reports the case of a 25-year-old woman with complaints of umbilical effusion and a mass. She was diagnosed with a urachal remnant and underwent urachal resection and reconstruction, with postoperative confirmation of favorable outcomes and the absence of microscopic hematuria. The intraoperative examination did not reveal any macroscopically clear luminal structure of the urachal resection margin. Subsequent histopathological analysis of the margin using hematoxylin and eosin staining was challenging, prompting the use of immunohistological staining with keratin AE1/AE3 antibody. The antibody did not stain the urachal resection margin, confirming the complete removal of urachal epithelial components. Our study findings suggest the utility of keratin AE1/AE3 staining for assessing urachal remnant margins and underscore the importance of thorough evaluation and complete resection of urachal remnant to prevent recurrence and mitigate the risk of urachal cancer, contributing to improved surgical outcomes and patient care.

Impact of common social habits on optical properties of lithium disilicate glass ceramic crowns: An in vitro study.

Statement of the problem: Patient stratifications considered the stability of color and treatment longevity are key success of restoration. Daily consumption of colored beverages, such as coffee, tea, and soft drinks, as well as the use of globally consumed materials, such as smokeless tobacco (ST), snuff, Khat, and Yerba mate, can change the color of restorative materials, such as lithium disilicate glass ceramics (LDGC). These changes can ultimately lead to treatment failure. Purpose: This in vitro study aimed to evaluate color changes, translucency, and opalescence of full anatomical LDGC crowns exposed to commonly used and potentially colorant solutions. Materials and methods: Ninety LDGC specimens/crowns were prepared and divided into nine groups according to immersion solution (control, Saudi Coffee, Cola, Khat, Yerba mate, Nescafe, ST Snuff, and Mixed Fruit Juice). The specimens were immersed in colorant solutions for 15 days with alternating twice daily at 37 °C. Color parameters were measured with a spectrophotometer and calculated using two backgrounds (black and white). Data were subjected to ANOVA followed by the Student t-test and Bonferroni test at a significant difference level (α = 0.05). Results: The greatest color change (ΔE*) among groups after immersion was observed in Yerba mate (7.6 ± 1.6). The mean difference of before and after staining within Yerba mate group was 3.14 ± 1.6 (p = 0.001). Translucency mean values of groups after immersion into staining media were ranging between 7.6 ± 1.2 and 9.1 ± 2, showing a slight decrease compared with pre-staining values but was not significantly different. Immersion in Mixed Fruit Juice significantly reduced opalescence (7.4 ± 1.9) compared to (8.8 ± 1.7) before staining. Conclusion: The findings confirm that appropriate user guidance helps to preserve both translucency and opalescence as well as prevent color changes. This can improve patient compliance and promote treatment longevity.

Automatic Annotation Diagnostic Framework for Nasopharyngeal Carcinoma via Pathology-Fidelity GAN and Prior-Driven Classification.

Non-keratinizing carcinoma is the most common subtype of nasopharyngeal carcinoma (NPC). Its poorly differentiated tumor cells and complex microenvironment present challenges to pathological diagnosis. AI-based pathological models have demonstrated potential in diagnosing NPC, but the reliance on costly manual annotation hinders development. To address the challenges, this paper proposes a deep learning-based framework for diagnosing NPC without manual annotation. The framework includes a novel unpaired generative network and a prior-driven image classification system. With pathology-fidelity constraints, the generative network achieves accurate digital staining from H&E to EBER images. The classification system leverages staining specificity and pathological prior knowledge to annotate training data automatically and to classify images for NPC diagnosis. This work used 232 cases for study. The experimental results show that the classification system reached a 99.59% accuracy in classifying EBER images, which closely matched the diagnostic results of pathologists. Utilizing PF-GAN as the backbone of the framework, the system attained a specificity of 0.8826 in generating EBER images, markedly outperforming that of other GANs (0.6137, 0.5815). Furthermore, the F1-Score of the framework for patch level diagnosis was 0.9143, exceeding those of fully supervised models (0.9103, 0.8777). To further validate its clinical efficacy, the framework was compared with experienced pathologists at the WSI level, showing comparable NPC diagnosis performance. This low-cost and precise diagnostic framework optimizes the early pathological diagnosis method for NPC and provides an innovative strategic direction for AI-based cancer diagnosis.

Clinicopathological Appearance of Epidermal Growth-Factor-Containing Fibulin-like Extracellular Matrix Protein 1 Deposition in the Lower Gastrointestinal Tract: An Autopsy-Based Study.

This study examined the patterns of epidermal growth-factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) deposition in the small intestine and colon to evaluate the association between the histopathological severity of EFEMP1 deposition and constipation and determine the colocalization of amyloid transthyretin (ATTR) and EFEMP1 deposits. In 40 older cases (≥80 years of age), EFEMP1 deposition in the small intestine initiated in the submucosal and subserous vessels, subserous interstitium, and serosa (early stage), progressing to the muscularis propria and peri-Auerbach plexus area (intermediate stage), and finally spreading diffusely to other areas, excluding the mucosa and muscularis mucosa (advanced stage). The colon had a similar pattern of progression. During the middle-to-advanced stages, amyloid formation was observed in some vascular and serous deposits. A subgroup of cases was identified in which EFEMP1 deposition was the only presumed cause of constipation. Additionally, we demonstrated the colocalization of ATTR and EFEMP1 deposition. Apple-green birefringence was detected under polarized light only in approximately one-half of the cases in the small intestine and one-third of the cases in the colon. These findings strongly suggest that EFEMP1 deposits are correlated with pathological conditions of the lower gastrointestinal tract. As the histopathological diagnosis using Congo red-stained specimens is challenging, the combined use of elastic fiber staining and EFEMP1 immunohistochemistry is recommended to identify EFEMP1 deposition.

Cryptosporidium Infections in Neonatal Calves on a Dairy Farm.

This study was conducted with the aim of the molecular identification of the protozoan parasite Cryptosporidium spp. in calves in the early stage of their development on a dairy farm in Eastern Slovakia. Twenty-five Holstein and Holstein cross calves were included in the study and monitored from their birth to the fifth week of life (1-5 weeks). Fresh fecal samples were collected from the same group of calves each week, except during the fourth week, and with the exception of Sample 8. All samples were analyzed using the Ziehl-Neelsen staining method and coproantigen was tested using the ELISA test as the screening method. Using the ELISA method, the highest incidence of cryptosporidiosis was observed in the second week of life of the calves, while the antigen was detected in 21 (91.6%) calves. Using the Ziehl-Neelsen staining method, the highest incidence was also observed in the second week, with an incidence rate of 62.5%. Positive isolates confirmed by the ELISA test were molecularly characterized. The species and subtypes of Cryptosporidium in the positive isolates were identified using PCR and the sequence analysis of the small subunit of the ribosomal 18S RNA (ssu rRNA) and the 60 kDa glycoprotein (gp60) genes of the parasite. The sequence analysis of 29 isolates at the 18S rRNA loci confirmed the presence of two species-Cryptosporidium parvum and Cryptosporidium ryanae. Out of 29 isolates, 25 were assigned to the species C. parvum, with the gp60 locus identified as genotype IIaA17G1R1. Among the individual animal groups, calves are the most common reservoirs of the C. parvum zoonotic species. This disease has significant public health implications as contact with livestock and their feces and working with barn manure are major sources of infection, not only for other animals but also for humans.

Phosphorus-Solubilizing Bacteria Enhance Cadmium Immobilization and Gene Expression in Wheat Roots to Reduce Cadmium Uptake.

The application of phosphorus-solubilizing bacteria is an effective method for increasing the available phosphorus content and inhibiting wheat uptake of heavy metals. However, further research is needed on the mechanism by which phosphorus-solubilizing bacteria inhibit cadmium (Cd) uptake in wheat roots and its impact on the expression of root-related genes. Here, the effects of strain Klebsiella aerogenes M2 on Cd absorption in wheat and the expression of root-related Cd detoxification and immobilization genes were determined. Compared with the control, strain M2 reduced (64.1-64.6%) Cd uptake by wheat roots. Cd fluorescence staining revealed that strain M2 blocked the entry of exogenous Cd into the root interior and enhanced the immobilization of Cd by cell walls. Forty-seven genes related to Cd detoxification, including genes encoding peroxidase, chalcone synthase, and naringenin 3-dioxygenase, were upregulated in the Cd+M2 treatment. Strain M2 enhanced the Cd resistance and detoxification activity of wheat roots through the regulation of flavonoid biosynthesis and antioxidant enzyme activity. Moreover, strain M2 regulated the expression of genes related to phenylalanine metabolism and the MAPK signaling pathway to enhance Cd immobilization in roots. These results provide a theoretical basis for the use of phosphorus-solubilizing bacteria to remediate Cd-contaminated fields and reduce Cd uptake in wheat.

Are Colpodella Species Pathogenic? Nutrient Uptake and Approaches to Diagnose Infections.

Colpodella species are free-living protists phylogenetically related to apicomplexans. Colpodella sp. have been detected in human and animal tissues, as well as in ticks and biting flies. The trophozoite and cyst stages of Colpodella species can be distinguished from stages of the prey Parabodo caudatus using Sam-Yellowe's trichrome staining. Colpodella species obtain nutrients by attaching to their prey, aspirating the prey's cytoplasmic contents into a posterior food vacuole and encysting. It is unclear whether both trophozoite and cyst stages are present in human and animal tissues. Molecular techniques have detected Colpodella species in human blood, cerebrospinal fluid, and in ticks and flies. However, no morphological information was reported to aid life-cycle stage identification of Colpodella species. This review discusses the increased reports of Colpodella species detection in animals and in arthropods and the need to identify stages present in human and animal tissues. We previously used Sam-Yellowe's trichrome staining to identify life-cycle stages of Colpodella sp. In this review, we examine the reports of Colpodella species detection in human and animal tissues to determine whether the identification of Colpodella species represents true infections or contaminations of samples collected during routine surveillance of piroplasm infections in animals and arthropods. This review also aims to provide insights regarding Colpodella, nutrient uptake, and the survival of Colpodella sp. within humans, animals, and arthropods, as well as whether the attachment of trophozoites to cells occurs in tissues leading to myzocytosis and endocytosis.

Optimization of spermatozoa analysis in mice: A comprehensive protocol.

Sperm quality is critical to predict reproductive alterations caused by immunological factors or toxicant agents. Yet, no detailed protocol has been published focusing on analyses of sperm parameters in mice. Our aim was to evaluate the most efficient diluent for mice sperm analyses and to optimize the sperm morphology classification, through the comparison of different staining methods. The diluents assessed were PBS (baseline), HTF, DMEM, 1 % BSA in PBS and 9 % skimmed powdered milk diluted in PBS. Spermatozoa were evaluated for vitality, motility, and morphology, smears were stained with Papanicolaou, HE, Giemsa, and Rapid staining. Sperm vitality and total motility reached better scores in milk based and DMEM diluents. HE raised up as an effective option since its combination with any of the diluents we tested, resulted in a fair staining, which was appropriated to evaluate mice spermatozoa. Finally, based on WHO manual, we have updated the current morphological classification for mice sperm, since we have detailed the head defects as well as included midpiece and tail defects on it. Taken together, we presented a useful, low cost, and reliable method to assess sperm morphology that could be employed worldwide by laboratories dedicated to study reproductive biology on mice model.

High-Speed Clearing and High-Resolution Staining for Analysis of Various Markers for Neurons and Vessels.

BACKGROUND: Tissue clearing enables deep imaging in various tissues by increasing the transparency of tissues, but there were limitations of immunostaining of the large-volume tissues such as the whole brain. METHODS: Here, we cleared and immune-stained whole mouse brain tissues using a novel clearing technique termed high-speed clearing and high-resolution staining (HCHS). We observed neural structures within the cleared brains using both a confocal microscope and a light-sheet fluorescence microscope (LSFM). The reconstructed 3D images were analyzed using a computational reconstruction algorithm. RESULTS: Various neural structures were well observed in three-dimensional (3D) images of the cleared brains from Gad-green fluorescent protein (GFP) mice and Thy 1-yellow fluorescent protein (YFP) mice. The intrinsic fluorescence signals of both transgenic mice were preserved after HCHS. In addition, large-scale 3D imaging of brains, immune-stained by the HCHS method using a mild detergent-based solution, allowed for the global topological analysis of several neuronal markers such as c-Fos, neuronal nuclear protein (NeuN), Microtubule-associated protein 2 (Map2), Tuj1, glial fibrillary acidic protein (GFAP), and tyrosine hydroxylase (TH) in various anatomical regions in the whole mouse brain tissues. Finally, through comparisons with various existing tissue clearing methodologies such as CUBIC, Visikol, and 3DISCO, it was confirmed that the HCHS methodology results in relatively less tissue deformation and higher fluorescence retention. CONCLUSION: In conclusion, the development of 3D imaging based on novel tissue-clearing techniques (HCHS) will enable detailed spatial analysis of neural and vascular networks present within the brain.

Advantages of laparoscopic segmentectomy of the liver using ICG fluorescent navigation by the negative staining method: A comparison with open procedure.

Aim: Laparoscopic segmentectomy (LS) using indocyanine green (ICG) fluorescence navigation with negative staining method has potential for performing accurate and safe anatomical excision. This study aimed to evaluate the significance of LS using ICG fluorescence navigation compared with open segmentectomy (OS). Methods: Eighty-seven patients who underwent anatomical segmentectomies were evaluated for OS (n = 44) and LS (n = 43). The Glissonean pedicle approach was performed using either extra- or intrahepatic method, depending on the location of segment in LS. After clamping pedicle, negative staining method was performed. Liver transection was done along intersegmental plane visualizing by overlay mode of ICG camera. Surgical outcomes were compared between two groups. Correlation between predicted resecting liver volume (PRLV) calculated using volumetry and actual resected liver volume (ARLV) was assessed in two groups. Results: Patients who underwent LS showed better outcomes in operative time, blood loss, and length of hospital stay. There were significantly fewer Grade II and Grade III or higher postoperative complications in LS group. Both values of AST (p < 0.001) and ALT (p < 0.001) on postoperative day 1 were significantly lower in LS group than in OS group. PRLV and ARLV were more strongly correlated in LS (r = 0.896) than in OS (r = 0.773). The difference between PRLV and ARLV was significantly lower in LS group than in OS group (p = 0.022), and this trend was particularly noticeable in posterosuperior segment (p = 0.008) than in anterolateral segment (p = 0.811). Conclusion: LS using ICG navigation allows precise resection and may contribute to safer short-term outcomes than OS, particularly in posterosuperior segment.

NOX4-mediated astrocyte ferroptosis in Alzheimer's disease.

This study investigates NADPH oxidase 4 (NOX4) involvement in iron-mediated astrocyte cell death in Alzheimer's Disease (AD) using single-cell sequencing data and transcriptomes. We analyzed AD single-cell RNA sequencing data, identified astrocyte marker genes, and explored biological processes in astrocytes. We integrated AD-related chip data with ferroptosis-related genes, highlighting NOX4. We validated NOX4's role in ferroptosis and AD in vitro and in vivo. Astrocyte marker genes were enriched in AD, emphasizing their role. NOX4 emerged as a crucial player in astrocytic ferroptosis in AD. Silencing NOX4 mitigated ferroptosis, improved cognition, reduced Aß and p-Tau levels, and alleviated mitochondrial abnormalities. NOX4 promotes astrocytic ferroptosis, underscoring its significance in AD progression.

Evaluation of the Genotoxic Effects of Grape Seed Extract and Marine Collagen Peptide on the Fibroblast Cell Line: An In Vitro Study.

Introduction Collagen plays a vital role in maintaining the structural integrity of dentin, and its modification with bioactive compounds can enhance its mechanical properties and bonding capabilities. Aim This study aimed to evaluate the genotoxic effects of grape seed extract (GSE) and marine collagen peptide (MCP) on dental pulp-derived primary cells. Methodology Human dental pulp stem cells were isolated, cultivated, and then treated with GSE and marine collagen peptides. DNA fragmentation was assessed using DAPI (4',6-diamidino-2-phenylindole) staining. Statistical analysis was performed using SPSS version 20 (IBM Corp., Armonk, NY, USA). Results The results showed that GSE exhibited a minimum level of cell death compared to marine collagen peptides. The viable cell count increased steadily over three days in all groups, with the control group showing the highest number of viable cells. The differences in viable cell count among the groups were statistically significant. Conclusion This study suggests that GSE and marine collagen peptides are highly biocompatible with dental pulp cells and could be considered for further clinical studies.

Spatial distribution pattern of immune cells is associated with patient prognosis in colorectal cancer.

BACKGROUND: The spatial context of tumor-infiltrating immune cells (TIICs) is important in predicting colorectal cancer (CRC) patients' clinical outcomes. However, the prognostic value of the TIIC spatial distribution is unknown. Thus, we aimed to investigate the association between TIICs in situ and patient prognosis in a large CRC sample. METHODS: We implemented multiplex immunohistochemistry staining technology in 190 CRC samples to quantify 14 TIIC subgroups in situ. To delineate the spatial relationship of TIICs to tumor cells, tissue slides were segmented into tumor cell and microenvironment compartments based on image recognition technology, and the distance between immune and tumor cells was calculated by implementing the computational pipeline phenoptr. RESULTS: MPO+ neutrophils and CD68+IDO1+ tumor-associated macrophages (TAMs) were enriched in the epithelial compartment, and myeloid lineage cells were located nearest to tumor cells. Except for CD68+CD163+ TAMs, other cells were all positively associated with favorable prognosis. The prognostic predictive power of TIICs was highly related to their distance to tumor cells. Unsupervised clustering analysis divided colorectal cancer into three subtypes with distinct prognostic outcomes, and correlation analysis revealed the synergy among B cells, CD68+IDO1+TAMs, and T lineage cells in producing an effective immune response. CONCLUSIONS: Our study suggests that the integration of spatial localization with TIIC abundance is important for comprehensive prognostic assessment.

Refining Complete Dentures: Optimizing Results with Enhanced Phonetics and Esthetics.

Tooth loss and subsequent complete denture rehabilitation can profoundly affect a patient's psychological and social well-being. Dentures play an important role in helping individuals regain a sense of normalcy as well as facilitating communication in today's appearance-conscious society. However, common issues with dentures include discomfort while chewing and dissatisfaction with esthetics and speech. Prosthetic rehabilitation for complete denture patients should aim not just at replacing missing teeth but at fully restoring masticatory functions and appearance. This article outlines a simple, economical, and esthetically pleasing approach to rehabilitating patients with complete dentures, particularly those with resorbed ridges and with difficulty in phonetics. The focus is on comprehensively restoring the patient's orofacial complex.

Deep learning-based hyperspectral technique identifies metastatic lymph nodes in oral squamous cell carcinoma-A pilot study.

AIMS: To establish a system based on hyperspectral imaging and deep learning for the detection of cancer cells in metastatic lymph nodes. MAIN METHODS: The continuous sections of metastatic lymph nodes from 45 oral squamous cell carcinoma (OSCC) patients were collected. An improved ResUNet algorithm was established for deep learning to analyze the spectral curve differences between cancer cells and lymphocytes, and that between tumor tissue and normal tissue. KEY FINDINGS: It was found that cancer cells, lymphocytes, and erythrocytes in the metastatic lymph nodes could be distinguished basing hyperspectral image, with overall accuracy (OA) as 87.30% and average accuracy (AA) as 85.46%. Cancerous area could be recognized by hyperspectral image and deep learning, and the average intersection over union (IOU) and accuracy were 0.6253 and 0.7692, respectively. SIGNIFICANCE: This study indicated that deep learning-based hyperspectral techniques can identify tumor tissue in OSCC metastatic lymph nodes, achieving high accuracy of pathological diagnosis, high work efficiency, and reducing work burden. But these are preliminary results limited to a small sample.

Isolation, Culture, and Phenotypic Analysis of Murine Lung Organoids.

Three-dimensional (3D) organoid cultures retain self-renewing stem cells that differentiate into multiple cell types that display spatial organization and functional key features, providing a highly physiological relevant system. Here we describe a strategy for the generation of 3D murine lung organoids derived from freshly isolated primary tracheal and distal lung epithelial stem cells. Isolated tracheas are subjected to enzymatic digestion to release the epithelial layer that is then dissociated into a single cell suspension for organoid culture. Lung epithelial cells are obtained from dissected lobes, which are applied to mechanical and enzymatic dissociation. After flow sorting, organoids are established from tracheal basal, secretory club, and alveolar type 2 cells in the defined conditioned medium that is required to sustain organoid growth and generate the differentiated cells. Multi-cell-type organoid co-culture replicates niches for distal epithelial stem cells to differentiate into bronchiolar and alveolar cell types. Established organoids can be fixed for wholemount staining and paraffin embedding, or passaged for further culture. Taken together, this protocol provides an efficient and validated approach to generate murine lung organoids, as well as a platform for further analysis.

Fluorescent labeling of micro/nanoplastics for biological applications with a focus on "true-to-life" tracking.

The increased environmental presence of micro-/nanoplastics (MNPLs) and the potential health risks associated with their exposure classify them as environmental pollutants with special environmental and health concerns. Consequently, there is an urgent need to investigate the potential risks associated with secondary MNPLs. In this context, using "true-to-life" MNPLs, resulting from the laboratory degradation of plastic goods, may be a sound approach. These non-commercial secondary MNPLs must be labeled to track their presence/journeys inside cells or organisms. Because the cell internalization of MNPLs is commonly analyzed using fluorescence techniques, the use of fluorescent dyes may be a sound method to label them. Five different compounds comprising two chemical dyes (Nile Red and Rhodamine-B), one optical brightener (Opticol), and two industrial dyes (Amarillo Luminoso and iDye PolyPink) were tested to determine their potential for such applications. Using commercial standards of polystyrene nanoplastics (PSNPLs) with an average size of 170 nm, different characteristics of the selected dyes such as the absence of impact on cell viability, specificity for plastic staining, no leaching, and lack of interference with other fluorochromes were analyzed. Based on the overall data obtained in the wide battery of assays performed, iDye PolyPink exhibited the most advantages, with respect to the other compounds, and was selected to effectively label "true-to-life" MNPLs. These advantages were confirmed using a proposed protocol, and labeling titanium-doped PETNPLs (obtained from the degradation of milk PET plastic bottles), as an example of "true-to-life" secondary NPLs. These results confirmed the usefulness of iDye PolyPink for labeling MNPLs and detecting cell internalization.

Diagnostic Maze: Malignant Phyllodes Tumor Mimicking Metaplastic Breast Carcinoma.

Phyllodes tumors (PTs) are uncommon breast tumors. They represent a spectrum of benign to malignant neoplasms. These erratic tumors have traits ranging from fibroadenomas on one end of the spectrum to sarcomas on the other end. The presentation of PT is variable, thus posing a diagnostic difficulty. Malignant PTs are often associated with recurrence, poor prognosis, and adverse disease outcomes. The recent genetic studies produced by genome sequencing offer information about the molecular pathophysiology of PT, aid in enhancing diagnostic precision, and suggest possible therapeutic targets in cases of malignant PT. Planning a treatment modality and prognostication requires meticulous histopathological sampling, as it relies on a correct histopathological diagnosis. There are no definitive guidelines for surgical management and targeted therapy for malignant PTs due to the rarity of these cases and very little available literature on these topics. Here in this article, we address a malignant PT in a 74-year-old female that presented as a breast mass mimicking metaplastic breast carcinoma histologically. This article also illuminates how immunohistochemistry plays a vital role in the diagnosis of this tumor.

Confocal Imaging of Seeds.

In flowering plants, proper seed development is achieved through the constant interplay of fertilization products, embryo and endosperm, and maternal tissues. Understanding such a complex biological process requires microscopy techniques able to unveil the seed internal morphological structure. Seed thickness and relatively low permeability make conventional tissue staining techniques impractical unless combined with time-consuming dissecting methods. Here, we describe two techniques to imaging the three-dimensional structure of Arabidopsis seeds by confocal laser scanning microscopy. Both procedures, while differing in their time of execution and resolution, are based on cell wall staining of seed tissues with fluorescent dyes.