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European Journal Of Histochemistry[JOURNAL]

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Spinosin ameliorates osteoarthritis through enhancing the Nrf2/HO-1 signaling pathway.

Lu P, Li S, Zhang C … +6 more , Jiang X, Xiang J, Xu H, Dong J, Wang K, Shi Y

Eur J Histochem · 2024 May · PMID 38779782 · Full text

Osteoarthritis (OA) is a common degenerative joint disease in the elderly, while oxidative stress-induced chondrocyte degeneration plays a key role in the pathologic progression of OA. One possible reason is that the exp... Osteoarthritis (OA) is a common degenerative joint disease in the elderly, while oxidative stress-induced chondrocyte degeneration plays a key role in the pathologic progression of OA. One possible reason is that the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), which acts as the intracellular defense factor against oxidative stress, is significantly inhibited in chondrocytes. Spinosin (SPI) is a potent Nrf2 agonist, but its effect on OA is still unknown. In this study, we found that SPI can alleviate tert-Butyl hydroperoxide (TBHP)-induced extracellular matrix degradation of chondrocytes. Additionally, SPI can effectively activate Nrf2, heme oxygenase-1 (HO-1), and NADPH quinone oxidoreductase 1 (NQO1) in chondrocytes under the TBHP environment. When Nrf2 was silenced by siRNA, the cartilage protective effect of SPI was also weakened. Finally, SPI showed good alleviative effects on OA in mice. Thus, SPI can ameliorate oxidative stress-induced chondrocyte dysfunction and exhibit a chondroprotective effect through activating the Nrf2/HO-1 pathway, which may provide a novel and promising option for the treatment of OA.

A candidate projective neuron type of the cerebellar cortex: the synarmotic neuron.

Flace P, Galletta D, Bizzoca A … +2 more , Gennarini G, Livrea P

Eur J Histochem · 2024 May · PMID 38766720 · Full text

Previous studies on the granular layer of the cerebellar cortex have revealed a wide distribution of different subpopulations of less-known large neuron types, called "non-traditional large neurons", which are distribute... Previous studies on the granular layer of the cerebellar cortex have revealed a wide distribution of different subpopulations of less-known large neuron types, called "non-traditional large neurons", which are distributed in three different zones of the granular layer. These neuron types are mainly involved in the formation of intrinsiccircuits inside the cerebellar cortex. A subpopulation of these neuron types is represented by the synarmotic neuron, which could play a projective role within the cerebellar circuitry. The synarmotic neuron cell body map within the internal zone of the granular layer or in the subjacent white substance. Furthermore, the axon crosses the granular layer and runs in the subcortical white substance, to reenter in an adjacent granular layer, associating two cortico-cerebellar regions of the same folium or of different folia, or could project to the intrinsic cerebellar nuclei. Therefore, along with the Purkinje neuron, the traditional projective neuron type of the cerebellar cortex, the synarmotic neuron is candidate to represent the second projective neuron type of the cerebellar cortex. Studies of chemical neuroanatomy evidenced a predominant inhibitory GABAergic nature of the synarmotic neuron, suggesting that it may mediate an inhibitory GABAergic output of cerebellar cortex within cortico-cortical interconnections or in projections towards intrinsic cerebellar nuclei. On this basis, the present minireview mainly focuses on the morphofunctional and neurochemical data of the synarmotic neuron, and explores its potential involvement in some forms of cerebellar ataxias.

Emodin improves renal fibrosis in chronic kidney disease by regulating mitochondrial homeostasis through the mediation of peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1α).

Feng L, Lin Z, Tang Z … +6 more , Zhu L, Xu S, Tan X, Wang X, Mai J, Tan Q

Eur J Histochem · 2024 May · PMID 38742403 · Full text

Chronic kidney disease (CKD) is a leading public health issue associated with high morbidity worldwide. However, there are only a few effective therapeutic strategies for CKD. Emodin, an anthraquinone compound from rhuba... Chronic kidney disease (CKD) is a leading public health issue associated with high morbidity worldwide. However, there are only a few effective therapeutic strategies for CKD. Emodin, an anthraquinone compound from rhubarb, can inhibit fibrosis in tissues and cells. Our study aims to investigate the antifibrotic effect of emodin and the underlying molecular mechanism. A unilateral ureteral obstruction (UUO)-induced rat model was established to evaluate the effect of emodin on renal fibrosis development. Hematoxylin and eosin staining, Masson's trichrome staining, and immunohistochemistry staining were performed to analyze histopathological changes and fibrotic features after emodin treatment. Subsequently, a transforming growth factor-beta 1 (TGF-β1)-induced cell model was used to assess the inhibition of emodin on cell fibrosis in vitro. Furthermore, Western blot analysis and real-time quantitative reverse transcription-polymerase chain reaction were performed to validate the regulatory mechanism of emodin on renal fibrosis progression. As a result, emodin significantly improved histopathological abnormalities in rats with UUO. The expression of fibrosis biomarkers and mitochondrial biogenesis-related proteins also decreased after emodin treatment. Moreover, emodin blocked TGF-β1-induced fibrotic phenotype, lipid accumulation, and mitochondrial homeostasis in NRK-52E cells. Conversely, peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1α) silencing significantly reversed these features in emodin-treated cells. Collectively, emodin plays an important role in regulating PGC-1α-mediated mitochondria function and energy homeostasis. This indicates that emodin exhibits great inhibition against renal fibrosis and acts as a promising inhibitor of CKD.

Erratum - LncRNA gadd7 promotes mitochondrial membrane potential decrease and apoptosis of alveolar type II epithelial cells by positively regulating MFN1 in an model of hyperoxia-induced acute lung injury.

Liu G, Yin C, Qian M … +3 more , Xiao X, Wu H, Fu F

Eur J Histochem · 2024 May · PMID 38713031 · Full text

This corrects the article published in European Journal of Histochemistry 2023;67:3535. doi: 10.4081/ejh.2023.3535. This corrects the article published in European Journal of Histochemistry 2023;67:3535. doi: 10.4081/ejh.2023.3535.

Pleural mesothelioma from fluoro-edenite exposure: PACAP and PAC1 receptor. A preliminary report.

Lombardo C, Maugeri G, D'Amico AG … +6 more , Broggi G, Caltabiano R, Filetti V, Matera S, D'Agata V, Loreto C

Eur J Histochem · 2024 May · PMID 38699968 · Full text

Pleural mesothelioma is a devastating malignancy primarily associated with asbestos exposure. However, emerging evidence suggests that exposure to fluoro-edenite fibers, a naturally occurring mineral fiber, can also lead... Pleural mesothelioma is a devastating malignancy primarily associated with asbestos exposure. However, emerging evidence suggests that exposure to fluoro-edenite fibers, a naturally occurring mineral fiber, can also lead to the development of pleural mesothelioma. In this study, based on the hypothesis that pituitary adenylate cyclase-activating polypeptide (PACAP) and PACAP-preferring receptor (PAC1R) expressions could be dysregulated in pleural mesothelioma samples and that they could potentially act as diagnostic or prognostic biomarkers, we aimed to investigate the immunohistochemical expression of PACAP and PAC1R in pleural biopsies from patients with pleural mesothelioma exposed to fluoro-edenite fibers. A total of 12 patients were included in this study, and their biopsies were processed for immunohistochemical analysis to evaluate the expression of PACAP and its receptor. The study revealed a correlation between the overexpression of PACAP and PAC1R and shorter overall survival in patients with malignant mesothelioma. These findings suggest that PACAP and PAC1R expression levels could serve as potential prognostic biomarkers for malignant mesothelioma. Furthermore, the immunohistochemical analysis of PACAP and PAC1R may provide valuable information for clinicians to guide therapeutic decisions and identify patients with poorer prognosis.

Pretreatment with interleukin-15 attenuates inflammation and apoptosis by inhibiting NF-κB signaling in sepsis-induced myocardial dysfunction.

He C, Yu Y, Wang F … +3 more , Li W, Ni H, Xiang M

Eur J Histochem · 2024 Apr · PMID 38686889 · Full text

Sepsis-induced myocardial dysfunction (SIMD) is associated with poor prognosis and increased mortality in patients with sepsis. Cytokines are important regulators of both the initiation and progression of sepsis. Interle... Sepsis-induced myocardial dysfunction (SIMD) is associated with poor prognosis and increased mortality in patients with sepsis. Cytokines are important regulators of both the initiation and progression of sepsis. Interleukin-15 (IL-15), a pro-inflammatory cytokine, has been linked to protective effects against myocardial infarction and myocarditis. However, the role of IL-15 in SIMD remains unclear. We established a mouse model of SIMD via cecal ligation puncture (CLP) surgery and a cell model of myocardial injury via lipopolysaccharide (LPS) stimulation. IL-15 expression was prominently upregulated in septic hearts as well as cardiomyocytes challenged with LPS. IL-15 pretreatment attenuated cardiac inflammation and cell apoptosis and improved cardiac function in the CLP model. Similar cardioprotective effects of IL-15 pretreatment were observed in vitro. As expected, IL-15 knockdown had the opposite effect on LPS-stimulated cardiomyocytes. Mechanistically, we found that IL-15 pretreatment reduced the expression of the pro-apoptotic proteins cleaved caspase-3 and Bax and upregulated the anti-apoptotic protein Bcl-2. RNA sequencing and Western blotting further confirmed that IL-15 pretreatment suppressed the activation of nuclear factor kappa B (NF-κB) signaling in mice with sepsis. Besides, the addition of NF-κB inhibitor can significantly attenuate cardiomyocyte apoptosis compared to the control findings. Our results suggest that IL-15 pretreatment attenuated the cardiac inflammatory responses and reduced cardiomyocyte apoptosis by partially inhibiting NF-κB signaling in vivo and in vitro, thereby improving cardiac function in mice with sepsis. These findings highlight a promising therapeutic strategy for SIMD.

Resveratrol mediates mitochondrial function through the sirtuin 3 pathway to improve abnormal metabolic remodeling in atrial fibrillation.

Cao Y, Cui L, Tuo S … +2 more , Liu H, Cui S

Eur J Histochem · 2024 Apr · PMID 38656259 · Full text

This study investigated the impact of resveratrol on abnormal metabolic remodeling in atrial fibrillation (AF) and explored potential molecular mechanisms. An AF cell model was established by high-frequency electrical st... This study investigated the impact of resveratrol on abnormal metabolic remodeling in atrial fibrillation (AF) and explored potential molecular mechanisms. An AF cell model was established by high-frequency electrical stimulation of HL-1 atrial muscle cells. Resveratrol concentrations were optimized using CCK-8 and flow cytometry. AF-induced increases in ROS and mitochondrial calcium, along with decreased adenosine triphosphate (ATP) and mitochondrial membrane potential, were observed. Resveratrol mitigated these changes and maintained normal mitochondrial morphology. Moreover, resveratrol acted through the SIRT3-dependent pathway, as evidenced by its ability to suppress AF-induced acetylation of key metabolic enzymes. SIRT3 overexpression controls acetylation modifications, suggesting its regulatory role. In conclusion, resveratrol's SIRT3-dependent pathway intervenes in AF-induced mitochondrial dysfunction, presenting a potential therapeutic avenue for AF-related metabolic disorders. This study sheds light on the role of resveratrol in mitigating AF-induced mitochondrial remodeling and highlights its potential as a novel treatment for AF.

Mast cells in meningiomas.

D'Amati A, Tamma R, Annese T … +2 more , Rizzi A, Ribatti D

Eur J Histochem · 2024 Apr · PMID 38634735 · Full text

Meningioma represents the most frequent tumor of the central nervous system (CNS). Correlations between the presence of mast cells (MCs) and grade or other histological features of meningioma are still debated. Our study... Meningioma represents the most frequent tumor of the central nervous system (CNS). Correlations between the presence of mast cells (MCs) and grade or other histological features of meningioma are still debated. Our study aimed to better understand the relationship between mast cells and meningiomas and to compare our results based on specific histological subtypes and novel 2021 CNS WHO grading system. We observed some differences as regards the number of MCs and meningioma grade. In low-grade (grade 1) meningiomas, MCs were observed in 7/22 cases, while they were consistently present in all eight high-grade cases (grade 2 and grade  3). Among the grade 1 meningiomas, we observed two "low-positive", two "intermediate-positive", and three "high-positive" cases. Among the group of high-grade meningiomas, the six cases grade 2 were considered as "low-positive", while the two grade 3 cases showed a higher number of MCs and were included in the "intermediate-positive" group. Even though with no statistical significance, due to the low number of cases, our results seem to confirm a sort of relationship between meningioma grading and the number of MCs, as demonstrated by the higher percentage of high-grade meningiomas showing MCs infiltrates, compared to low-grade meningiomas.

Walking a thin line between fixation and epitope binding - characterization of antigen retrieval methods suitable for eosinophil and HSV-2 staining in formalin-fixed female reproductive tissue.

Wadephul LM, Arndts K, Katawa G … +4 more , Dietlmeier E, Horsnell W, Hoerauf A, Ritter M

Eur J Histochem · 2024 Apr · PMID 38624064 · Full text

Antibody-based fluorescence analysis of female reproductive tissues in research of sexually transmitted diseases allows for an in-depth understanding of protein localization, interactions, and pathogenesis. However, in m... Antibody-based fluorescence analysis of female reproductive tissues in research of sexually transmitted diseases allows for an in-depth understanding of protein localization, interactions, and pathogenesis. However, in many cases, cryosectioning is not compatible with biosafety regulations; at all times, exposure of lab personnel and the public to potentially harmful pathogens from biological infectious material must be avoided; thus, formaldehyde fixation is essential. Due to formaldehyde's cross-linking properties, protein detection with antibodies can be impeded. To allow effective epitope binding during immunofluorescence of formalin-fixed paraffin-embedded vaginal tissue, we investigated two antigen retrieval methods. We tested these methods regarding their suitability for automated image analysis, facilitating reproducible quantitative microscopic data acquisition in sexually transmitted disease research. Heat-based retrieval at 80°C in citrate buffer proved to increase antibody binding to eosinophil protein and HSV-2 visibly and tissue morphology best, and was the most efficient for sample processing and quantitative analysis.

Histological analysis of the dermal and hypodermal layers of the face and correlation with high-frequency 24 MHz ultrasonography and elastosonography.

Veronese S, Costa E, Portuese A … +2 more , Ossanna R, Sbarbati A

Eur J Histochem · 2024 Apr · PMID 38619113 · Full text

Knowledge of the structure of the face is of fundamental importance. In fact, the face is treated in many areas of medicine, from dermatology, to maxillofacial surgery, to otorhinolaryngology, to ophthalmology, etc. and... Knowledge of the structure of the face is of fundamental importance. In fact, the face is treated in many areas of medicine, from dermatology, to maxillofacial surgery, to otorhinolaryngology, to ophthalmology, etc. and anti-aging aesthetic treatments, and those for the resolution of blemishes are on the increase. For ethical reasons it is not possible to take biopsy samples for facial analysis in the aesthetic field. The main aim of this study was to demonstrate that a high-resolution bimodal ultrasound examination, combined with elastosonography, could be a valid tool for pre-treatment morphological evaluation. To achieve this goal, skin samples were taken from the forehead, zygomatic area, nasolabial fold, upper and lower lip from cadavers to histologically characterize their structure. Subsequently, these same areas were evaluated in vivo using conventional B-mode ultrasound with a 24 MHz high-frequency probe, and elastosonography. The data obtained with the different techniques were compared, in order to state that modern ultrasound techniques can provide similar histological information. The analysis showed that the superficial hypodermis presented a different shape and structure in the different areas, with the exception of the areas of the upper and lower lip, which appeared similar. With aging, the forehead and zygomatic area showed a volumetric increase in the superficial hypodermic layer, while the lip showed non-structural changes. The morphology of the nasolabial fold remained unchanged. When it is not possible to perform histological investigations on the face, to understand its characteristics and dynamics, ultrasound with a 24 MHz probe would seem to be the most suitable method, while elastosonography could be a valid method for evaluating the stiffness of the structural components.

Atorvastatin reduces calcification in valve interstitial cells the NF-κB signalling pathway by promoting Atg5-mediated autophagy.

Chen M, Liu S

Eur J Histochem · 2024 Apr · PMID 38619020 · Full text

Aortic valve calcification (AVC) is a common cardiovascular disease and a risk factor for sudden death. However, the potential mechanisms and effective therapeutic drugs need to be explored. Atorvastatin is a statin that... Aortic valve calcification (AVC) is a common cardiovascular disease and a risk factor for sudden death. However, the potential mechanisms and effective therapeutic drugs need to be explored. Atorvastatin is a statin that can effectively prevent cardiovascular events by lowering cholesterol levels. However, whether atorvastatin can inhibit AVC by reducing low-density lipoprotein (LDL) and its possible mechanism of action require further exploration. In the current study, we constructed an in vitro AVC model by inducing calcification of the valve interstitial cells. We found that atorvastatin significantly inhibited osteogenic differentiation, reduced the deposition of calcium nodules in valve interstitial cells, and enhanced autophagy in calcified valve interstitial cells, manifested by increased expression levels of the autophagy proteins Atg5 and LC3B-II/I and the formation of smooth autophagic flow. Atorvastatin inhibited the NF-κB signalling pathway and the expression of inflammatory factors mediated by NF-κB in calcified valve interstitial cells. The activation of the NF-κB signalling pathway led to the reversal of atorvastatin's effect on enhancing autophagy and alleviating valve interstitial cell calcification. In conclusion, atorvastatin inhibited the NF-κB signalling pathway by upregulating autophagy, thereby alleviating valve interstitial cell calcification, which was conducive to improving AVC.

Style head in Apocynaceae: a very complex secretory activity performed by one tissue.

Maximo D, Demarco D

Eur J Histochem · 2024 Apr · PMID 38568208 · Full text

Nuptial glands are very diverse and associated with different pollination mechanisms. The greater the specificity in the pollen transfer mechanism from anther to stigma, the greater the morphological elaboration of flowe... Nuptial glands are very diverse and associated with different pollination mechanisms. The greater the specificity in the pollen transfer mechanism from anther to stigma, the greater the morphological elaboration of flowers and functional complexity of the nuptial glands. In Apocynaceae, pollination mechanisms reached an extreme specificity, a fact that was only possible due to an extreme morphological synorganization and a profusion of floral glands. Although these glands are of different types, the vast majority have secretory cells only in the epidermis. In general, these epidermal cells produce many different compounds at the same time, and previous studies have demonstrated that in the style head, the functional complexity of epidermis has become even greater. Four types of style head are found in the family, which have different degrees of functional complexity in relation to the secretion produced and pollen dispersal mechanism. The secretion is fluid in types I, II and III, and the pollen is dispersed and adhered to the pollinator by the secretion produced by the style head. In type IV, the secretion hardens and acquires a specific shape, moulded by the spatial constraints of the adjacent floral organs. This evolutionary alteration is accompanied by changes in the structure and arrangement of the secretory cells, as well as in pollen aggregation and position of stigma. Histochemical analysis has shown that the secretion is mixed and highly complex, especially in the style head type IV, where the secretion, called translator, is formed by a rigid central portion, which adheres to the pollinator, and two caudicles that attach to two pollinia. The translator has a distinct composition in its different parts. Further studies are needed to answer the new questions that have arisen from the discovery of this highly functional complexity of the secretory tissue.

Senescence-associated ß-galactosidase staining over the lifespan differs in a short- and a long-lived fish species.

Schöfer S, Laffer S, Kirchberger S … +7 more , Kothmayer M, Löhnert R, Ebner EE, Weipoltshammer K, Distel M, Pusch O, Schöfer C

Eur J Histochem · 2024 Feb · PMID 38568207 · Full text

During the aging process, cells can enter cellular senescence, a state in which cells leave the cell cycle but remain viable. This mechanism is thought to protect tissues from propagation of damaged cells and the number... During the aging process, cells can enter cellular senescence, a state in which cells leave the cell cycle but remain viable. This mechanism is thought to protect tissues from propagation of damaged cells and the number of senescent cells has been shown to increase with age. The speed of aging determines the lifespan of a species and it varies significantly in different species. To assess the progress of cellular senescence during lifetime, we performed a comparative longitudinal study using histochemical detection of the senescence-associated beta-galactosidase as senescence marker to map the staining patterns in organs of the long-lived zebrafish and the short-lived turquoise killifish using light- and electron microscopy. We compared age stages corresponding to human stages of newborn, childhood, adolescence, adult and old age. We found tissue-specific but conserved signal patterns with respect to organ distribution. However, we found dramatic differences in the onset of tissue staining. The stained zebrafish organs show little to no signal at newborn age followed by a gradual increase in signal intensity, whereas the organs of the short-lived killifish show an early onset of staining already at newborn stage, which remains conspicuous at all age stages. The most prominent signal was found in liver, intestine, kidney and heart, with the latter showing the most prominent interspecies divergence in onset of staining and in staining intensity. In addition, we found staining predominantly in epithelial cells, some of which are post-mitotic, such as the intestinal epithelial lining. We hypothesize that the association of the strong and early-onset signal pattern in the short-lived killifish is consistent with a protective mechanism in a fast growing species. Furthermore, we believe that staining in post-mitotic cells may play a role in maintaining tissue integrity, suggesting different roles for cellular senescence during life.

The Feulgen reaction at the electron microscopy level.

Biggiogera M

Eur J Histochem · 2024 Feb · PMID 38568205 · Full text

The Feulgen reaction has been the first specific method for detecting DNA available at light microscopy since 1924. However, a similar specific method was proposed for electron microscopy only 50 years later. Here, we di... The Feulgen reaction has been the first specific method for detecting DNA available at light microscopy since 1924. However, a similar specific method was proposed for electron microscopy only 50 years later. Here, we discuss the problems encountered in finding the electrondense reagent capable of taking advantage of the extremely high resolution offered by electron microscopy as well as some applications of the method.

The role of galanin in the progression and prognosis of colorectal cancer: the unfinished story.

Kmiec Z, Kieżun J, Krazinski BE … +2 more , Kwiatkowski P, Godlewski J

Eur J Histochem · 2024 Mar · PMID 38568200 · Full text

The paper presents a summary of immunohistochemical (IHC) and biochemical investigations on the presence of galanin (Gal), one of the neuropeptides abundant in the enteric nervous systems, and three types of its receptor... The paper presents a summary of immunohistochemical (IHC) and biochemical investigations on the presence of galanin (Gal), one of the neuropeptides abundant in the enteric nervous systems, and three types of its receptors (GalR1-3) in colorectal cancer (CRC) tissue and non-involved colon wall and their associations with clinical-pathological data of the CRC patients. We were the first to morphologically demonstrate the presence of endogenous Gal in CRC sections and measure its content in homogenates of tumor tissue and dissected compartments of unchanged colon wall. The prominent atrophy of myenteric plexuses displaying Gal immunoreactivity (Gal-Ir) located close to the tumor invasion was found to be accompanied by higher Gal content in the tumor-adjacent muscularis externa than in tumor-distant tissue. In further studies for the first time, we demonstrated by the IHC technique the presence of the GalR1-3 receptors in the CRC tumors and the colon mucosa and found that higher GalR3-Ir in the tumor tissue correlated with longer overall survival of CRC patients. Furthermore, we discovered that lower GalR1 expression in submucosal plexuses located near the tumor correlated with a better prognosis in patients with CRC. These findings suggest that GalR1 could be considered as a novel therapeutic target in CRC. In conclusion, our morphological investigations provided novel data documenting the involvement of Gal and its receptors in the progression of CRC and showed the usefulness of the IHC technique for the prognosis of CRC patients.

The Feulgen reaction: from pink-magenta to rainbow fluorescent at the Maffo Vialli's School of Histochemistry.

Mazzini G

Eur J Histochem · 2024 Feb · PMID 38389457 · Full text

For over a century, Palazzo Botta (Palace Botta) has housed the University of Pavia's Biomedical Institutes. Illustrious scientists have conducted research and taught at this Palace, making significant contributions to t... For over a century, Palazzo Botta (Palace Botta) has housed the University of Pavia's Biomedical Institutes. Illustrious scientists have conducted research and taught at this Palace, making significant contributions to the advancement of natural, biological, and medical science. Among them, Camillo Golgi received the Nobel Prize for discovering the so-called "black reaction." Following Golgi, the Palace continued to be a hub for the development of methodologies and reactions aimed at detecting and quantifying biological components. Maffo Vialli (in the Golgi stream) was the first to establish a Histochemistry Research Group, which began in the naturalistic field and later expanded to the biomedical area. Among the many histochemical studies initiated in the Palace, the Feulgen reaction undoubtedly played a significant role. This reaction, developed R. Feulgen and H. Rossenbeck in 1924, had significant international implications: numerous researchers then contributed to define its fine chemical details, which remained the subject of study for years, resulting in a massive international scientific literature. The Pavia School of Histochemistry also contributed to the evolution and application of this method, which has become a true benchmark in quantitative histochemistry. Giovanni Prenna and the CNR Centre for Histochemistry made significant contributions, as they were already focused on fluorescence cytochemistry. The Pavia researchers made significant contributions to the development of methodology and, in particular, instrumentation; the evolution of the latter resulted in the emergence of flow cytometry and an ever-increasing family of fluorescent probes, which somewhat overshadowed the Feulgen reaction for DNA quantification. The advent of monoclonal antibodies then contributed to the final explosion of flow cytometry in clinical application, almost making young neophytes forget that its roots date back to Feulgen.

1954-2024: 70 years of histochemical research with the European Journal of Histochemistry.

Pellicciari C, Biggiogera M, Malatesta M

Eur J Histochem · 2024 Jan · PMID 38285131 · Full text

This Editorial celebrates the 70th anniversary of the European Journal of Histochemistry since its foundation as Rivista di Istochimica Normale e Patologica, and introduces a Special Collection of selected articles on th... This Editorial celebrates the 70th anniversary of the European Journal of Histochemistry since its foundation as Rivista di Istochimica Normale e Patologica, and introduces a Special Collection of selected articles on the application of the histochemical approach for investigating cell biological features and processes in animals and plants, and under diseased conditions. The year 2024 is a special one for histochemists, as 100 years ago J.W. Robert Feulgen and H. Rossenbeck introduced the histochemical procedure for the specific stoichiometric staining of DNA in histological samples: to commemorate this influential publication, three papers in the present issue are devoted to the application of the Feulgen reaction at light and electron microscopy, and in cytometry.

Giving bodies to ghosts: locating molecules in the very place where they exert their biological roles.

Capanna E, Redi C

Eur J Histochem · 2024 Jan · PMID 38285084 · Full text

This paper reviews some of the goals of our investigations published over the years on Rivista di Istochimica Normale e Patologica, Basic and Applied Histochemistry, and the European Journal of Histochemistry - EJH. In a... This paper reviews some of the goals of our investigations published over the years on Rivista di Istochimica Normale e Patologica, Basic and Applied Histochemistry, and the European Journal of Histochemistry - EJH. In a series of papers, we published some of the basic cytochemical features of the sperm cytodifferentiation process for the first time. This was a conceptual and practical prerequisite to the in situ quantitative evaluation of sperm DNA content. We showed that the discrepancy between the expected 1:2 ratio when comparing sperm versus somatic cell DNA content (sperm DNA content is always far low from the theoretical value) is due to DNA losses caused by the hydrochloric treatment entailed by the Feulgen reaction. The knowledge of the specific losses that occur during the various steps of the Feulgen reaction has allowed us to use it critically in Genome Size studies to highlight: - sperm aneuploidy in chromosomally derived subfertility; - the broad variability range of Mammalian genome sizes; - that termites are roaches (after decades of discussion on this topic). In addition, in a seminal paper on human oocytes, we showed (by transmission electron microscopy) a specific chromatin and cytoplasmic organization (both essential for further embryo development) linked to oocyte maturation arrest, a datum quite relevant to treating unmet therapeutic needs in human and veterinary reproduction.

Lectins as versatile tools to explore cellular glycosylation.

Brooks S

Eur J Histochem · 2024 Jan · PMID 38285057 · Full text

Lectins are naturally occurring carbohydrate-binding proteins that are ubiquitous in nature and highly selective for their, often incompletely characterised, binding partners. From their discovery in the late 1880s to th... Lectins are naturally occurring carbohydrate-binding proteins that are ubiquitous in nature and highly selective for their, often incompletely characterised, binding partners. From their discovery in the late 1880s to the present day, they have provided a broad palette of versatile tools for exploring the glycosylation of cells and tissues and for uncovering the myriad functions of glycosylation in biological systems. The technique of lectin histochemistry, used to map the glycosylation of tissues, has been instrumental in revealing the changing profile of cellular glycosylation in development, health and disease. It has been especially enlightening in revealing fundamental alterations in cellular glycosylation that accompany cancer development and metastasis, and has facilitated the identification of glycosylated biomarkers that can predict prognosis and may have utility in development of early detection and screening, Moreover, it has led to insights into the functional role of glycosylation in healthy tissues and in the processes underlying disease. Recent advances in biotechnology mean that our understanding of the precise binding partners of lectins is improving and an ever-wider range of lectins are available, including recombinant human lectins and lectins with enhanced, engineered properties. Moreover, use of traditional histochemistry to support a broad range of cutting-edge technologies and the development of high throughout microarray platforms opens the way for ever more sophisticated mapping - and understanding - of the glycome.

Investigation of seasonal changes in lipid synthesis and metabolism-related genes in the oviduct of Chinese brown frog (<em>Rana dybowskii</em>).

Wang Y, Liu Y, Wang Y … +5 more , Zhang A, Xie W, Zhang H, Weng Q, Xu M

Eur J Histochem · 2023 Dec · PMID 38116875 · Full text

A peculiar physiological characteristic of the Chinese brown frog (Rana dybowskii) is that its oviduct dilates during pre-brumation rather than during the breeding season. This research aimed to examine the expression of... A peculiar physiological characteristic of the Chinese brown frog (Rana dybowskii) is that its oviduct dilates during pre-brumation rather than during the breeding season. This research aimed to examine the expression of genes connected with lipid synthesis and metabolism in the oviduct of R. dybowskii during both the breeding season and pre-brumation. We observed significant changes in the weight and size of the oviduct between the breeding season and pre-brumation. Furthermore, compared to the breeding season, pre-brumation exhibited significantly lower triglyceride content and a marked increase in free fatty acid content. Immunohistochemical results revealed the spatial distribution of triglyceride synthase (Dgat1), triglyceride hydrolase (Lpl and Hsl), fatty acid synthase (Fasn), and fatty acid oxidases (Cpt1a, Acadl, and Hadh) in oviductal glandular cells and epithelial cells during both the breeding season and pre-brumation. While the mRNA levels of triglycerides and free fatty acid synthesis genes (dgat1 and fasn) did not show a significant difference between the breeding season and pre-brumation, the mRNA levels of genes involved in triglycerides and free fatty acid metabolism (lpl, cpt1a, acadl, acox and hadh) were considerably higher during pre-brumation. Furthermore, the R. dybowskii oviduct's transcriptomic and metabolomic data confirmed differential expression of genes and metabolites enriched in lipid metabolism signaling pathways during both the breeding season and pre-brumation. Overall, these results suggest that alterations in lipid synthesis and metabolism during pre-brumation may potentially influence the expanding size of the oviduct, contributing to the successful overwintering of R. dybowskii.
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