Histological Events Research Articles

Morphogenesis <i>in vitro</i> in calluses of lavender <i>Lavandula angustifolia</i> Mill.: histological aspects

Histological events occurring in the calluses of Lavandula angustifolia Mill. at the initial stages of in vitro culture (1 passage) were described for the first time. It was found that the non-morphogenic callus is mainly represented by parenchymal tissue with few morphogenetic foci, mostly degenerated. In morphogenic calluses the morphogenesis pathways such as de novo organogenesis and indirect in vitro somatic embryogenesis have been identified and multiple developing morphogenetic foci have been noted also. The question of the realization of the pluri- and totipotency properties of callus cells in vitro is discussed. The histological data can be used in choosing the direction of application of regenerants obtained from calluses of this valuable essential oil and medicinal plant in various cell technologies.

Efficacy and Safety of Pembrolizumab plus Axitinib combination for Metastatic Renal Cell Carcinoma in a Real-World Scenario: Data From the Prospective ProPAXI Study

BackgroundPembrolizumab/Axitinib combination is approved as first-line therapy in mRCC. The aim of this study is to evaluate outcomes of PAXI combo in the real-world in Italy. MethodsThis is a prospective study including patients diagnosed with mRCC who received combination as first-line therapy in recruiting Italian Centers. Data about patient characteristics, safety and outcome were collected. Results170 pts have been treated from December 2020 to September 2023. The majority had clear-cell histology (83%). Sarcomatoid feature was present in 33%of available cases. About one half of patients (55%) had synchronous metastasis. In 58% of cases nephrectomy was performed, of which 27% were cytoreductive and 4% were deferred nephrectomies. Lung metastases were identified in 106 patients (62%), bone and liver involvement in 66 and 29 patients (38.8% and 17.1%) respectively. Stratifying by IMDC criteria, 32 patients (18.8%) were at favorable-risk, 106 (62.4%) at intermediate-risk, and 32 (18.8%) at poor-risk. At time of analysis, treatment was ongoing in 49% of patients. Progression occurred in 45% of patients. Median PFS was 19.2 months (95% CI: 15-NR). With a median follow-up of 19.3 months (range 1.3-34.5), at 24-months and 36-months landmark analysis 62% (95% CI, 53-70) and 58% (95% CI, 47-69) of treated patients are still alive respectively. Disease control rate was achieved in 84.6% of patients: 4.3% reached a complete response, 52% had a partial response and 28.8% a stable disease. Primary progression was observed in 15.3% of patients. In the multivariate analysis, the prognostic significance of age ≥ 65 years, non-clear cell histology, IMDC score, and adverse events and gender interaction as predictors of worse OS were confirmed. ConclusionThis is the first available prospective study on first-line Pembrolizumab/Axitinib combination in real world scenario. Our findings support the effectiveness and safety of first-line this combination in mRCC and reveal that gender emerged as a prognostic factor in relation to the occurrence of adverse events.

Atherosclerotic Plaque Epigenetic Age Acceleration Predicts a Poor Prognosis and Is Associated With Endothelial-to-Mesenchymal Transition in Humans.

Epigenetic age estimators (clocks) are predictive of human mortality risk. However, it is not yet known whether the epigenetic age of atherosclerotic plaques is predictive for the risk of cardiovascular events. Whole-genome DNA methylation of human carotid atherosclerotic plaques (n=485) and of blood (n=93) from the Athero-Express endarterectomy cohort was used to calculate epigenetic age acceleration (EAA). EAA was linked to clinical characteristics, plaque histology, and future cardiovascular events (n=136). We studied whole-genome DNA methylation and bulk and single-cell transcriptomics to uncover molecular mechanisms of plaque EAA. We experimentally confirmed our in silico findings using in vitro experiments in primary human coronary endothelial cells. Male and female patients with severe atherosclerosis had a median chronological age of 69 years. The median epigenetic age was 65 years in females (median EAA, -2.2 [interquartile range, -4.3 to 2.2] years) and 68 years in males (median EAA, -0.3 [interquartile range, -2.9 to 3.8] years). Patients with diabetes and a high body mass index had higher plaque EAA. Increased EAA of plaque predicted future events in a 3-year follow-up in a Cox regression model (univariate hazard ratio, 1.7; P=0.0034) and adjusted multivariate model (hazard ratio, 1.56; P=0.02). Plaque EAA predicted outcome independent of blood EAA (hazard ratio, 1.3; P=0.018) and of plaque hemorrhage (hazard ratio, 1.7; P=0.02). Single-cell RNA sequencing in plaque samples from 46 patients in the same cohort revealed smooth muscle and endothelial cells as important cell types in plaque EAA. Endothelial-to-mesenchymal transition was associated with EAA, which was experimentally confirmed by TGFβ-triggered endothelial-to-mesenchymal transition inducing rapid epigenetic aging in coronary endothelial cells. Plaque EAA is a strong and independent marker of poor outcome in patients with severe atherosclerosis. Plaque EAA was linked to mesenchymal endothelial and smooth muscle cells. Endothelial-to-mesenchymal transition was associated with EAA, which was experimentally validated. Epigenetic aging mechanisms may provide new targets for treatments that reduce atherosclerosis complications.

Mucosal adhesion phenomenon after maxillary sinus floor elevation: A preclinical study.

To describe the histological events that occur after maxillary sinus floor elevation when the elevated and undetached sinus mucosa are in close proximity or in contact with each other. From 76 rabbits, 152 elevated maxillary sinuses were analyzed histologically. Sites without adhesions were classified as "No proximity," whereas the adhesion stages were divided into "Proximity," "Fusion," and "Synechia stages." The width of the pseudostratified columnar epithelium and the distance between the two layers of the elevated and undetached sinus mucosae were measured at various standardized positions. Thirty-one sites presenting with adhesions were found. Twelve sites were in the proximity stage," presenting cilia of the two epithelial layers that were shortened and interlinked within the mucous context. Hyperactivity of the goblet cells was also observed. In the other cases, the hyperplastic epithelium showed attempts to reach the contralateral mucosa. The 15 "fusion stage" sites presented regions with epithelial cells of the two mucosal layers that penetrated each other. Four sites presented "synechiae stages," represented by bridges of connective tissue connecting the two lamina propria. Close proximity or tight contact between the elevated and undetached mucosa adhering to the bone walls might occur after maxillary sinus floor elevation. This induced hyperplasia of the epithelial cells and adhesion of the two layers until synechiae formation.

Early posttransplant reductions in club cell secretory protein associate with future risk for chronic allograft dysfunction in lung recipients: results from a multicenter study

Chronic lung allograft dysfunction (CLAD) increases morbidity and mortality for lung transplant recipients. Club cell secretory protein (CCSP), produced by airway club cells, is reduced in the bronchoalveolar lavage fluid (BALF) of lung recipients with CLAD. We sought to understand the relationship between BALF CCSP and early posttransplant allograft injury and determine if early posttransplant BALF CCSP reductions indicate later CLAD risk. We quantified CCSP and total protein in 1606 BALF samples collected over the first posttransplant year from 392 adult lung recipients at 5 centers. Generalized estimating equation models were used to examine the correlation of allograft histology or infection events with protein-normalized BALF CCSP. We performed multivariable Cox regression to determine the association between a time-dependent binary indicator of normalized BALF CCSP level below the median in the first posttransplant year and development of probable CLAD. Normalized BALF CCSP concentrations were 19% to 48% lower among samples corresponding to histological allograft injury as compared with healthy samples. Patients who experienced any occurrence of a normalized BALF CCSP level below the median over the first posttransplant year had a significant increase in probable CLAD risk independent of other factors previously linked to CLAD (adjusted hazard ratio 1.95; p=0.035). We discovered a threshold for reduced BALF CCSP to discriminate future CLAD risk; supporting the utility of BALF CCSP as a tool for early posttransplant risk stratification. Additionally, our finding that low CCSP associates with future CLAD underscores a role for club cell injury in CLAD pathobiology.

The frequency of dysplastic branching crypts in colorectal polypoid tubular adenomas.

Dysplastic crypt branching (DCB) was recently found in ulcerative colitis-associated dysplasia. The aim was to assess the frequency and the branching phenotype of DCB in polypoid colorectal tubular adenomas (TA). A total of 3956 DCB were found in the 139 TA: 98% were in asymmetric branching (DCAB) and the remaining 2% in symmetric branching (DCSB). A linear correlation was found between DCB frequency and the increasing digital size in TA (p < .05). Using a digital ruler, adenomas were divided into small TA (<5 mm) and larger TA (≥5 mm). The difference between the frequency of DCB in small TA (n=75) vs. larger TA (n=64), was significant (p < .05). DCB frequency was not influenced by age, gender or TA localization. In the normal colorectal mucosa (≈2 m2 ), only occasional CSB is found and no CAB. And yet, multiple DCB (mean 16.7 DCB), mostly DCAB, was found in small TA, occupying <5 mm of the mucosal area. In larger TA, as many as 42.1 DCB (mean), mostly DCAB, occurred in merely 7.8 mm (mean) of the colon mucosa. Thus it is suggestedthat DCB is a standard histologic element of TA. The natural expansion of the adenomatous tissue in larger TA appears to be follow on from newly produced, mostly DCAB, by DCSB and by the accumulation of their dysplastic offspring's progenies. The findings strongly suggest that DCB is a central microstructure in the histological events unfolding in polypoid colorectal TA.

Histological and molecular responses of Vigna angularis to Uromyces vignae infection

BackgroundTo advance the understanding of adzuki bean (Vigna angularis) resistance to infection with the rust-causing fungus Uromyces vignae (Uv), we comprehensively analyzed histological events and the transcriptome of Uv-infected adzuki bean.ResultsCompared with the susceptible cv. Baoqinghong (BQH), the resistant cv. QH1 showed inhibition of uredospore germination and substomatal vesicle development, intense autofluorescence of cells around the infection site, and cell wall deposit formation in response to Uv infection. In cv. QH1, gene set enrichment analysis (GSEA) showed enrichment of chitin catabolic processes and responses to biotic stimuli at 24 h post-inoculation (hpi) and cell wall modification and structural constituent of cytoskeleton at 48 hpi. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated enrichment of WRKY transcription factors (TFs), the calcium binding protein cml, and hydroquinone glucosyltransferase at both 24 and 48 hpi. In total, 1992 and 557 differentially expressed genes (DEGs) were identified at 24 and 48 hpi, respectively. Cell surface pattern-recognition receptors (PRRs), WRKY TFs, defense-associated pathogenesis-related (PR) proteins, and lignin and antimicrobial phenolic compound biosynthesis were significantly induced. Finally, we detected the chitinase (CHI) and phenylalanine ammonia-lyase (PAL) activity were higher in QH1 and increased much earlier than in BQH.ConclusionIn cv. QH1, cell-surface PRRs rapidly recognize Uv invasion and activate the corresponding TFs to increase the transcription of defense-related genes and corresponding enzymatic activities to prevent fungal development and spread in host tissues.

Liver and spleen volumes are associated with prognosis of compensated and decompensated cirrhosis and parallel its natural history.

ObjectiveCirrhosis is characterized by the complex interplay among biological, histological and haemodynamic events. Liver and spleen remodelling occur throughout its natural history, but the prognostic role of these volumetric changes is unclear. We evaluated the relationship between volumetric changes assessed by multidetector computerised tomography (MDCT) and landmark features of cirrhosis.MethodsWe included consecutive cirrhotic patients who underwent liver transplantation (LT) or hepatocellular carcinoma (HCC) resection in whom dynamic MDCT was available. Different volumetric indices were calculated. Fibrosis was evaluated by the collagen proportional area and Laennec sub‐stages. Correlation and logistic regression analysis were performed to explore associations of volumetric indexes and fibrosis with key prognostic features across the clinical stages of cirrhosis.Results185 patients were included (146 LT; 39 HCC); the predominant aetiology was viral hepatitis (51.35%); 65.9% had decompensated disease and 85.08% clinically significant portal hypertension (CSPH). The standardised liver volume and liver‐spleen volume ratio negatively correlated with Model for End‐stage Liver Disease (MELD), albumin and hepatic venous pressure gradient (HVPG) and were significantly lower in decompensated patients. The liver segmental volume ratio (segments I–III/segments IV–VIII) best captured the characteristic features of the compensated phase, showing a positive correlation with HVPG and a good discrimination between patients with and without CSPH and varices. Volumetric changes and fibrosis severity were independently associated with key prognostic events, with no association between these two parameters.ConclusionsLiver and spleen volumetric indices evolve differently along the natural history of cirrhosis and are associated with key prognostic factors in each phase, regardless of fibrosis severity and portal hypertension.

Embryology of the Abdominal Wall and Associated Malformations-AReview.

In humans, the incidence of congenital defects of the intraembryonic celom and its associated structures has increased over recent decades. Surgical treatment of abdominal and diaphragmatic malformations resulting in congenital hernia requires deep knowledge of ventral body closure and the separation of the primary body cavities during embryogenesis. The correct development of both structures requires the coordinated and fine-tuned synergy of different anlagen, including a set of molecules governing those processes. They have mainly been investigated in a range of vertebrate species (e.g., mouse, birds, and fish), but studies of embryogenesis in humans are rather rare because samples are seldom available. Therefore, we have to deal with a large body of conflicting data concerning the formation of the abdominal wall and the etiology of diaphragmatic defects. This review summarizes the current state of knowledge and focuses on the histological and molecular events leading to the establishment of the abdominal and thoracic cavities in several vertebrate species. In chronological order, we start with the onset of gastrulation, continue with the establishment of the three-dimensional body shape, and end with the partition of body cavities. We also discuss well-known human etiologies.

Basic fibroblast growth factor uniquely stimulates quiescent vascular smooth muscle cells and induces proliferation and dedifferentiation.

Blood vessels normally remain stable over the long term. However, in atherosclerosis, vascular cells leave the quiescent state and enter an activated state. Here, we investigated the factors that trigger the breakage of the quiescent state by screening growth factors and cytokines using a vascular smooth muscle cell (SMC) line and an endothelial cell (EC) line. Despite known functions of the tested factors, only basic fibroblast growth factor (bFGF) was identified as a potent trigger of quiescence breakage in SMCs, but not ECs. bFGF disrupted tight SMC-monolayers and caused morphological changes, proliferation, and dedifferentiation. Human primary SMCs, but not ECs, also showed similar results. Aberrant SMC proliferation is a critical histological event in atherosclerosis. We, thus, provide further insights into the role of bFGF in vascular pathobiology.

Advances in the Study of the Mechanisms of Physiological Root Resorption in Deciduous Teeth.

Physiological root resorption of deciduous teeth is a complex physiological process that is essential for the normal replacement of deciduous teeth and permanent teeth in clinical practice, but its importance is often overlooked due to the presence of permanent teeth. This physiological process includes not only the resorption of hard tissues of deciduous teeth, such as dentin and cementum, but also the elimination of soft tissues, such as pulp and periodontal ligament (PDL). However, the mechanisms of physiological root resorption are not yet clear. In this article, the advances of research on the mechanisms related to physiological root resorption will be reviewed in two main aspects: hard tissues and soft tissues of deciduous teeth, specifically in relation to the effects of inflammatory microenvironment and mechanical stress on the resorption of hard tissues, the repair of hard tissues, and the elimination and the histological events of soft tissues.

Transcriptome Profiling Reveals Role of MicroRNAs and Their Targeted Genes during Adventitious Root Formation in Dark-Pretreated Micro-Shoot Cuttings of Tetraploid Robinia pseudoacacia L.

Tetraploid Robinia pseudoacacia L. is a difficult-to-root species, and is vegetatively propagated through stem cuttings. Limited information is available regarding the adventitious root (AR) formation of dark-pretreated micro-shoot cuttings. Moreover, the role of specific miRNAs and their targeted genes during dark-pretreated AR formation under in vitro conditions has never been revealed. The dark pretreatment has successfully promoted and stimulated adventitious rooting signaling-related genes in tissue-cultured stem cuttings with the application of auxin (0.2 mg L−1 IBA). Histological analysis was performed for AR formation at 0, 12, 36, 48, and 72 h after excision (HAE) of the cuttings. The first histological events were observed at 36 HAE in the dark-pretreated cuttings; however, no cellular activities were observed in the control cuttings. In addition, the present study aimed to uncover the role of differentially expressed (DE) microRNAs (miRNAs) and their targeted genes during adventitious root formation using the lower portion (1–1.5 cm) of tetraploid R. pseudoacacia L. micro-shoot cuttings. The samples were analyzed using Illumina high-throughput sequencing technology for the identification of miRNAs at the mentioned time points. Seven DE miRNA libraries were constructed and sequenced. The DE number of 81, 162, 153, 154, 41, 9, and 77 miRNAs were upregulated, whereas 67, 98, 84, 116, 19, 16, and 93 miRNAs were downregulated in the following comparisons of the libraries: 0-vs-12, 0-vs-36, 0-vs-48, 0-vs-72, 12-vs-36, 36-vs-48, and 48-vs-72, respectively. Furthermore, we depicted an association between ten miRNAs (novel-m0778-3p, miR6135e.2-5p, miR477-3p, miR4416c-5p, miR946d, miR398b, miR389a-3p, novel m0068-5p, novel-m0650-3p, and novel-m0560-3p) and important target genes (auxin response factor-3, gretchen hagen-9, scarecrow-like-1, squamosa promoter-binding protein-like-12, small auxin upregulated RNA-70, binding protein-9, vacuolar invertase-1, starch synthase-3, sucrose synthase-3, probable starch synthase-3, cell wall invertase-4, and trehalose phosphatase synthase-5), all of which play a role in plant hormone signaling and starch and sucrose metabolism pathways. The quantitative polymerase chain reaction (qRT-PCR) was used to validate the relative expression of these miRNAs and their targeted genes. These results provide novel insights and a foundation for further studies to elucidate the molecular factors and processes controlling AR formation in woody plants.

Abstract P1-02-08: Comparison of clinical features and outcomes for pleomorphic invasive lobular carcinoma vs. non-pleomorphic invasive lobular carcinoma

Abstract Background: Invasive lobular carcinoma (ILC) is the second most common invasive breast cancer histology. Within ILC histology, there are multiple subtypes. The most common subtype is “classic” characterized by small single files of cells invading the stroma. Another subtype of clinical interest is “pleomorphic” which is characterized by larger cells with varied nuclear appearance. Rare and not well understood, pleomorphic ILC is traditionally viewed as an aggressive form of ILC. Studies have shown a relatively consistent association between pleomorphic ILC and certain high risk features such as advanced stage, HER2 positivity, and lymph node involvement. Survival outcome data comparing pleomorphic ILC to non-pleomorphic ILC has yielded varied results. Methods: A retrospective study of a large institutional database was conducted to characterize patients with ILC treated from 2004-2017. Patient and disease characteristics were recorded, including disease staging, biomarker profile, specific ILC histology, treatment details, and recurrence events. Findings were analyzed and overall survival (OS) and recurrence free survival (RFS) were compared between pleomorphic and non-pleomorphic ILC cohorts. Propensity score matching was also completed to account for confounding variables. Results: A total of 691 patients who had surgery were studied (mean age 61.6 ± 11.6 years old, 86.9% Caucasian, 98.3% with endocrine therapy), including 100 (14.4%) with pleomorphic ILC and 591 (85.5%) with non-pleomorphic ILC. Compared to non-pleomorphic ILC, pleomorphic ILC was less likely to be estrogen receptor (ER) positive (94% vs. 98%, p=0.004), more likely to be HER2 positive (12% vs. 7%, p=0.07), more likely to be grade 3 (33% vs. 2%, p&amp;lt;0.001), less likely to be diagnosed at a lower T stage (Stage I 38% vs. 58%, Stages II-III 61% vs. 41%, p&amp;lt;0.001), and less likely to be diagnosed with axillary lymph node involvement (node positivity 48.5% vs. 66%, p&amp;lt;0.001). Mastectomy rates were 65% in pleomorphic ILC and 52% in non-pleomorphic ILC (p=0.022). Patients with pleomorphic ILC were more likely to receive chemotherapy (66% vs. 36%, p&amp;lt;0.001). Pleomorphic ILC patients were also more likely to receive HER2-directed therapy (12% vs. 5.8%, p=0.02). Median follow-up was 6 years. There was no significant difference in OS and RFS between cohorts before matching for age, BMI, stage, surgery type, and chemotherapy. Although not statistically significant, pleomorphic ILC appears to have better RFS and OS between propensity score matched 91 pairs (10-year RFS: 79% vs. 71.5%, p=0.27 and 10-year OS: 86% vs. 76%, p=0.13). Exploratory analysis also suggests that multifocal pleomorphic ILC is associated with worse RFS and OS compared to unifocal pleomorphic ILC. Conclusions: Pleomorphic ILC was found to have more advanced disease at presentation in the breast but not in lymph nodes. At our institution, it was treated with more chemotherapy compared to non-pleomorphic ILC, likely related to higher grade, lower ER-positivity, and higher HER2-positivity. After propensity score matching, pleomorphic ILC appears to have better RFS and OS compared to non-pleomorphic ILC. This was particularly noticeable three years after surgery and may reflect more aggressive management. These data reveal some unexpected trends that challenge the notion of pleomorphic ILC having worse outcomes compared with non-pleomorphic ILC. This highlights the need for additional studies to better understand multiple aspects of ILC and its subtypes. Citation Format: Matthew D. Wright, Marcus S. Dempster, Ayat ElSherif, Daniela Cocco, Stephanie A. Valente, Hong Li, Megan L. Kruse. Comparison of clinical features and outcomes for pleomorphic invasive lobular carcinoma vs. non-pleomorphic invasive lobular carcinoma [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-02-08.

The timing of auditory sensory deficits in Norrie disease has implications for therapeutic intervention.

Norrie disease is caused by mutation of the NDP gene, presenting as congenital blindness followed by later onset of hearing loss. Protecting patients from hearing loss is critical for maintaining their quality of life. This study aimed to understand the onset of pathology in cochlear structure and function. By investigating patients and juvenile Ndp-mutant mice, we elucidated the sequence of onset of physiological changes (in auditory brainstem responses, distortion product otoacoustic emissions, endocochlear potential, blood-labyrinth barrier integrity) and determined the cellular, histological, and ultrastructural events leading to hearing loss. We found that cochlear vascular pathology occurs earlier than previously reported and precedes sensorineural hearing loss. The work defines a disease mechanism whereby early malformation of the cochlear microvasculature precedes loss of vessel integrity and decline of endocochlear potential, leading to hearing loss and hair cell death while sparing spiral ganglion cells. This provides essential information on events defining the optimal therapeutic window and indicates that early intervention is needed. In an era of advancing gene therapy and small-molecule technologies, this study establishes Ndp-mutant mice as a platform to test such interventions and has important implications for understanding the progression of hearing loss in Norrie disease.

Microscopical observations on the regenerating tail of tsinling dwarf skink (Scincella tsinlingensis)

Although the key steps of tail regeneration in lizards are well understood, further investigations involving skinks can provide the field of regeneration research with additional information. In order to characterize the cytoarchitecture of tail regeneration in Scincella tsinlingensis, an endemic species in China, its histological events and growth trends are investigated. The rate of tail regeneration varies with the season: it proceeds faster in summer and autumn than it does in winter and spring. Tail regeneration of S. tsinlingensis is summarized as wound healing, blastema formation, cell differentiation and tail growth, which can be subdivided into seven stages. Wound healing following tail loss, begins with an obvious outgrowth undergoing re-epithelialization. Numerous proliferating mesenchymal-like cells aggregate near the distal end of the severed spinal cord to form the blastema. The expanding blastema is invaded by blood vessels, nerves and ependyma. A cartilaginous skeleton is formed around the ependymal tube and the muscle starts to differentiate. The keratinization of epidermis coincides with scale formation. Pigmentation eventually occurs in the regenerated tail. Tail regeneration in S. tsinlingensis is an epimorphic kind of regeneration that is also known as blastema-mediated. Structure and composition of the regenerated tail, including its cytoarchitecture, represent a conserved pattern of regeneration also known from other lizards.

Histological and cytological study on meristematic nodule induction and shoot organogenesis in Paeonia ostii ‘Feng Dan’

This is the first report concerning the sequence of histological and cytological events occurring during organogenesis from cotyledon-derived meristematic nodules (MNs) in Paeonia ostii ‘Feng Dan’. Sections were made and studies were carried out with dissecting microscope, light microscope, scanning electron microscopy (SEM) and transmission electron microscopy observation. Histological studies revealed a complex developmental process of morphogenesis including five stages: (1) callus originated from cell division in both cambial and cortical regions; type I—yellow compact callus with densely arranged clumps was identified as embryogenic callus. (2) pre-nodular structure consisted of organization center (a central area of vascularization surrounded by meristematic cell layers) and an epidermis-like layer; (3) independent MNs comprised of organization center, a cortical-like area of parenchymatous cells and an epidermal-like area; (4) nodular clusters displayed vigorously internal meristematic cell division and generated a relative movement towards the nodules periphery, establishing vascular connection with primordia; (5) successive new elongated shoots with axillary bud primordia were developed. SEM observations showed three types of extracellular matrix, a smooth membranous layer, fibrillar structures and granular mucilage-like secretions on embryogenic callus, demonstrating its dynamic morphological changes. Ultrastructural analysis revealed striking changes of chloroplast morphology and starch content during MNs morphogenesis. This study allows a better understanding of in vitro regeneration via MN culture and provides references for protocol optimization and genetic transformation.

New Insights for Consummate Diagnosis and Management of Oral Submucous Fibrosis Using Reactive and Reparative Fibrotic Parameter Derived Algorithm.

Objective:Reproducibility of qualitative changes in histopathological diagnosis involving narrow variation is often challenging. This study aims to characterize the histological fibrotic events in detail so as to derive an in-depth multiparametric algorithm with individually quantified histological parameters for effective monitoring of the. disease process in oral submucous fibrosis and for potential therapeutic targets for early intervention.Methods:Formalin fixed paraffin embedded (FFPE) blocks of oral submucous fibrosis (OSMF), were taken and sections were stained with Hematoxylin & Eosin stain and Masson Trichrome stain. Photomicrographs were assessed for various morphometric parameters with Image J software version 1.8. Linear Regression was used to model the relationship using Inflammatory Cell Count, Extent of Inflammation collagen stained area, Epithelial thickness integrated density of collagen, MVPA, Area, Perimeter, were taken as variables.Result:Inflammatory cell count and the extent of inflammation also decreased with increasing grades of OSMF. Collagen proportionate area, integrated collagen density and epithelial thickness were compared among different grades of OSMF. Grade IV OSMF had greatest mean collagen proportionate area , highest integrated collagen density and lowest epithelial thickness when compared to other grades of OSMF. Linear regression model revealed smaller variation between Grade I to Grade II. Whereas Grade II to Grade IV exhibited larger variation suggestive of increased growth rate and all the coefficients were found to lie within 95% confidence limitsConclusion:Diagnostic algorithm with multiparametric regression model were derived and combinatorial therapeutic approaches have been suggested for more effective management of oral submucous fibrosis

Effects of diazinon on olfactory epithelium and genes related to olfactory signal transduction in Caspian roach, Rutilus caspicus

Olfaction in fishes is biologically-essential chemoreceptors. It provokes important behaviors for the survival of fish such as feeding, reproduction, avoiding predator and intraspecific communication. Pesticides can affect the olfactory epithelium and genes related to olfactory transduction and impair olfactory function. The present study aims to determine how toxicity of diazinon impacts olfaction of Caspian roach, Rutilus caspicus in histological and gene levels. We exposed fish (0.7 ± 0.05 g in weight) to 0, 1, 2 and 3 mg L-1 diazinon (60 fish in each 3- replicate 60-L glass tank) for 7 days in freshwater. Diazinon concentrations were chosen based on reports on its concentration in surface waters. Mortality rate, histological events of olfactory epithelium, and genes expression at the 7th day after diazinon exposure included a significant decrease in the number of olfactory receptor cells (ORCs), while increased goblet cells (GCs). In addition, G­protein αi (Gpαi) was significantly upregulated, whereas calmodulin-dependent kinase II α (CaMKIIα) was significantly downregulated after 7 days as compared to the control group. These results indicate that diazinon can impair olfactory function through effect on the olfactory epithelium and olfactory signal transduction pathways in the Caspian roach fingerlings.

Real-life use of BRAF-V600E mutation analysis in thyroid nodule fine needle aspiration: consequences on clinical decision-making.

This study aimed to evaluate the real-life use of BRAF-V600E mutation analysis in washout liquid from thyroid nodule fine needle aspiration (FNA), and the consequences of genetic result on clinical decision-making. We retrospectively considered subjects tested for BRAF-V600E among those attending the Endocrinology Unit of Modena for FNA between 2014 and 2018. Washing fluid was collected together with cytological sample and stored at -20 °C. If the clinician deemed it necessary, the sample was thawed, DNA extracted, and genetic test performed by high-resolution melting technique. We collected data on cytology according to the Italian Consensus for the cytological classification of thyroid nodules, type of surgery (when performed), histology, and adverse events. Out of 7112 subjects submitted to FNA, BRAF analysis was requested for 683 (9.6%). Overall, 896 nodules were analyzed: 74% were indeterminate at cytology, mainly TIR3A (low risk). Twenty-two nodules were mutant (BRAF+). Only 2% of indeterminate, mainly TIR3B, were BRAF+. Based on final histological diagnosis, BRAF test had high specificity (100%) but poor sensitivity (21%), also in indeterminate nodules. Mutant subjects underwent more extensive surgery compared to wild type (p = 0.000), with frequent prophylactic central lymph node dissection. One third had local metastases. Higher prevalence of hypoparathyroidism was found in BRAF+ compared to wild type (p = 0.018). The analysis of BRAF-V600E outside of gene panels has low sensitivity, especially in indeterminate nodules, and a positive result could lead to more extensive surgery with greater risk of hypoparathyroidism and questionable clinical utility.

Bone tissue and histological and molecular events during development of the long bones

The bones are of mesenchymal or ectomesenchymal origin, form the skeleton of most vertebrates, and are essential for locomotion and organ protection. As a living tissue they are highly vascularized and remodelled throughout life to maintain intact. Bones consist of osteocytes entrapped in a mineralized extracellular matrix, and via their elaborated network of cytoplasmic processes they do not only communicate with each other but also with the cells on the bone surface (bone lining cells). Bone tissue develops through a series of fine-tuned processes, and there are two modes of bone formation, referred to either as intramembranous or endochondral ossification. In intramembranous ossification, bones develop directly from condensations of mesenchymal cells, and the flat bones of the skull, the clavicles and the perichondral bone cuff develop via this process. The bones of the axial (ribs and vertebrae) and the appendicular skeleton (e.g. upper and lower limbs) form through endochondral ossification where mesenchyme turns into a cartilaginous intermediate with the shape of the future skeletal element that is gradually replaced by bone. Endochondral ossification occurs in all vertebrate taxa and its onset involves differentiation of the chondrocytes, mineralization of the extracellular cartilage matrix and vascularization of the intermediate, followed by disintegration and resorption of the cartilage, bone formation, and finally – after complete ossification of the cartilage model – the establishment of an avascular articular cartilage. The epiphyseal growth plate regulates the longitudinal growth of the bones, achieved by a balanced proliferation and elimination of chondrocytes, and the question whether the late hypertrophic chondrocytes die or transform into osteogenic cells is still being hotly debated. The complex processes leading to endochondral ossification have been studied for over a century, and this review aims to give an overview of the histological and molecular events, arising from the long bones’ (e.g. femur, tibia) development. The fate of the hypertrophic chondrocytes will be discussed in the light of new findings obtained from cell tracking studies.