Round Nuclei Research Articles

PATH-18. MORE UNUSUAL FEATURES IN H3 K27M-MUTANT GLIOMAS

Abstract Diffuse midline glioma, H3 K27M-altered, WHO grade 4 is an infiltrating midline glioma; however, H3 K27M mutation can also be seen in midline circumscribed, non-infiltrating gliomas and rarely in non-midline tumors. We report 2 unusual examples. A 37-year-old female developed a right posterior frontal infiltrating glioma; resection showed infiltrative tumor with brisk mitotic activity, elevated MIB-1 (20%), negative p53 expression, ATRX retention, but no necrosis or microvascular proliferation. Molecular workup showed neither IDH1/2/TERT promoter mutations or co-deletion 1p, 19q; further testing showed gain of whole chromosome 7 + loss of whole chromosome 10, meeting molecular criteria for glioblastoma. Recurrent tumor 2 years later in the same area prompted re-resection again of infiltrating tumor devoid of necrosis or microvascular proliferation; molecular testing now revealed a H3F3A (p.K28M) mutation. Both tumors showed protein expression by H3 K27M IHC+. She succumbed post-therapy at 3 ½ years. A 30-year-old woman developed a circumscribed, non-enhancing third ventricular mass; resection demonstrated circumscribed, non-infiltrating glioneuronal tumor with monotonous rounded tumor nuclei, synaptophysin IHC+, negative GFAP, H3 K27M IHC+, and low MIB-1. Molecular testing identified alterations in H3F3A p. K28M and FGFR1 p.N546K. She was treated, but lost to follow-up, until she re-presented 5 years later with hemorrhagic left frontal lobe tumor contiguous with the midline, with dorsal midbrain and 4th ventricular involvement. After resection, tumor now showed infiltrative features, necrosis and microvascular proliferation, diffuse GFAP IHC+, retention of nuclear ATRX, p53 (20%), MIB-1 (22%), and H3 K27M IHC+. New tumor had no FGFR1 alteration or PDGFRA amplification to suggest radiation-induced tumor, instead manifesting PTPRZ1-MET fusion. She survives at 5 years. These cases offer new insights into survival of a non-midline example and transformation of a circumscribed, non-infiltrating tumor to secondary infiltrating H3 K27-altered tumor with new molecular alterations, possibly representing second independent tumor development.

MODL-15. ESTABLISHMENT OF ORGANOID MODELS OF PATIENT-DERIVED GH-PRODUCING PITUITARY ADENOMA

Abstract BACKGROUND Growth hormone-producing pituitary adenomas (GHomas) reveal unfavorable clinical course due to systemic complications such as excessive sweating, impaired glucose tolerance, and hypertension, in addition to its characteristic facial appearance. For GHoma, pharmacological treatments including somatostatin analogs and dopamine agonists are widely performed, however, a subset of case reveals dismal prognosis. We need to elucidate molecular mechanisms contributing to GHoma growth, however, experimental models of GHoma such as cell line and animal model are quietly limited. OBJECTIVE In recent years, organoid models using 3D-culture technologies have gained attention as useful research models for various types of cancers. This study aims to establish a novel organoid model derived from GHoma patients. Using established GHoma organoid model, we analyzed their histological characteristics. METHODS Surgical specimens from five cases of GHoma were finely dissected to approximately 1mm and embedded in Matrigel basement membrane matrix. The specimens were then cultured under 37°C conditions in a 5% CO2 incubator using a medium supplemented with various growth factors. After 28 days of culture, formalin-fixed paraffin-embedded sections were prepared, and the histological similarity with patient tumors was examined. RESULTS Hematoxylin and eosin staining showed that organoid models consisted of acidophilic, granular cytoplasm and small round nuclei, maintaining a cell morphology similar with the original tumor. Immunohistochemical staining revealed positive expression of growth hormone, Pit-1, and CAM5.2, which are specific markers for GHoma. CONCLUSIONS We successfully established a patient-derived organoid model of GHoma that retains histological and endocrinological characteristics. We believe that this model could be a valuable research model for elucidating the molecular mechanisms of GHoma and developing novel treatment approaches targeting important molecular abnormalities. Additionally, in clinical settings, this model might be useful for predicting the responsiveness to drug treatment and selecting appropriate therapies for individual patients.

The cranial apparatus glands of the canthariphilous Pyrochroa coccinea (Coleoptera: Pyrochroidae: Pyrochroinae), and their implications in sexual behaviour

Some Pyrochroidae species are known as “canthariphilous” for their attraction to cantharidin (CTD), a toxic terpene with anti-predatory effects, produced in nature by only two beetle families (Meloidae and Oedemeridae). It has been demonstrated that males of Neopyrochroa flabellata ingesting CTD are positively selected by females. Indeed, the compound is re-emitted from a glandular cranial apparatus as secretions that are licked up by females during courtship behaviour, inducing copulation. Herein, we provide the first description of the glands associated to the cranial apparatus of male Pyrochroinae using the European species Pyrochroa coccinea as a model. Morphological analyses show that the cranial apparatus consists of a concave pit lined with short setae retaining secretions emitted through numerous glandular pores. Ultrastructural investigations reveal the presence of two different class 3 glands (Gl.A and Gl.B), intermixed at the level of the pit but exhibiting distinct features. Gl.A are mainly characterised by short conducting canals, rounded nuclei and electrondense vesicles while Gl.B are characterised by long conducting canals, irregular nuclei, vesicles containing a particulate substance and a multifolded plasma membrane. Observations of sexual behaviour are also reported for P. coccinea and compared to N. flabellata, confirming the involvement of cranial apparatus secretions in courtship behaviour.

A fully automated Lab-on-a-Disc platform integrated a high-speed triggered siphon valve for PBMCs extraction

Human Peripheral Blood Mononuclear Cells (PBMCs) are isolated from peripheral blood and identified as any blood cell with a round nucleus that exhibits immune responses and undergoes immunophenotypic changes upon exposure to various pathophysiological stimuli. Obtaining high-recovery and clinical-grade PBMCs without decreasing cell viability and causing stress is crucial for disease diagnosis and successful immunotherapy. However, traditional manual PBMCs extraction methods rely on manual intervention with less recovery rate and reliability. In this study, we introduced a novel and efficient strategy for the fully automated extraction of PBMCs based on a Lab-on-a-Disk (LoaD) platform. The centrifugal chip used percoll as density gradient media (DGM) for separation and extraction on account of the density difference of cells in whole blood, without labeling and any additional extra cellular filtration or cell lysis steps. Above all, we proposed a high-speed triggered siphon valve, which was closed under the speed of cell sedimentation and subsequently opened by increasing speed to complete the extraction of PBMCs. It can avoid the problem that previous siphon valves rely on unstable hydrophilic surface treatment and prime under low/zero speed conditions. With valves and the clock channel integrated on the chip, users can achieve fully automated collection of PBMCs. Compared with the clinical laboratory results, the recovery rate of extracted PBMCs was 80 %. The experimental results prove that the high-speed triggered siphon valve improves the extraction efficiency of PBMCs. The robust chips, which are not only simple to manufacture and assemble but also stable and reliable to use, have great potential in biomedical and clinical applications.

THU517 Aggressive Cushing’s: A Rare Case Of Pancreatic ACTHoma In A Young Female

Abstract Disclosure: O.C. Reyes: None. A. Sison: None. M. Villa: None. D. Dimayuga: None. Introduction: Adrenocorticotropic hormone-producing pancreatic neuroendocrine neoplasm (ACTHoma) is a rare type of pancreatic neuroendocrine tumor that causes ectopic adrenocorticotropic hormone syndrome. Cushingoid manifestations and metabolic abnormalities can occur rapidly. High index of suspicion is required for early diagnosis. Case: A 31-year-old Filipino female presented with facial and ankle swelling that occurred 6 weeks before the visit. She also had acne, easy bruising, amenorrhea and frequent mood swings. There was reported weight gain of 3 kg a month prior. She had no history of hypertension and diabetes, and had no unusual family history. At the time of admission, vital signs were blood pressure of 130/80mmHg, pulse of 92/min, respiratory rate 20/min and body temperature of 37.2C. Her body mass index was 28kg/m2. She had muscle atrophy of the upper and lower extremities, facial flushing, posterior cervical fat pad, abdominal violaceous striae, and skin pigmentation. Her external genitalia was normal, Tanner Stage 5 consistent with adult pattern. Neurologic exam was unremarkable. She was examined for Cushing’s syndrome. On initial tests, glucose was 406 mg/dl, HbA1C 10.7%, thyroid stimulating hormone 0.122 (NV: 0.55-4.78), Free T3 0.77 (NV: 2.3-4.2), Free T4 0.58 (NV: 0.89-1.76). Creatinine was 0.86 mg/dL, sodium was 140 mEq/L, potassium was 2.1 mEq/L showing hypokalemia. There was leukocytosis on her blood count, white blood cells 15,310/mm3 (neutrophils 94%, lymphocytes 3%). In the low-dose dexamethasone suppression test (DST), the serum cortisol concentration was 991.03nmol/L. Her adrenocorticotrophic hormone (ACTH) level was elevated at 577pg/mL. After high-dose DST, the serum cortisol concentration was > 1750 nmol/L. No tumor was observed on pituitary magnetic resonance imaging. On abdominal computed tomography (CT), a large, lobulated, heterogeneously-enhancing mass arising from the pancreatic body and tail measuring 7.5cm x 10cm x 10.9cm was seen. The mass had infiltrated the gastric fundus, and encased the splenic artery. Multiple non-calcified subcentimeter pulmonary nodules were observed on chest CT. CT-guided biopsy of the pancreatic mass was done. On microscopic examination, numerous atypical cells arranged with small clusters and sheets and scattered singly were found in the tumor. The cells had small to enlarged, hyperchromatic, ovoid to round nuclei and scant to ample delicate cytoplasm. On immunohistochemical staining, the tumor cells were stained positive for synaptophysin and chromogranin A, which are both neuroendocrine markers, as well as ACTH. However, the patient died prior to any definitive management. Conclusion: ACTHoma is a very rare disease that causes hypercortisolemia. Reporting of similar cases will provide insight into its clinical features, immunohistochemical characteristics, diagnosis, therapy, and prognosis. Presentation: Thursday, June 15, 2023

SAT203 Parathyroid Adenoma Masquerading As Parathyroid Carcinoma

Abstract Disclosure: M. Fariduddin: None. A. Lupieri: None. H.L. Calero: None. A. Rubenfeld: None. Y. Eisenberg: None. Case: 22 y/o pregnant female (5 weeks gestation) presented to the endocrinology clinic after a hospital admission for nausea, vomiting, constipation, fatigue and joint pain with labs showing calcium (corrected) of 14 mg/dL and PTH of 1200 pg/ml. Labs in the clinic showed calcium at 16 mg/dL and PTH 1500 pg/ml. She was re-admitted for IV fluids, calcitonin and zoledronic acid. Severe hypercalcemia, young age, and extremely high PTH levels were highly suggestive of parathyroid cancer. CT of neck showed a 2 cm right inferior parathyroid mass abutting the esophagus with concern for esophageal invasion with no evident plane between the mass and the esophagus. No cervical adenopathy was noted. Patient in conjunction with her obstetrician made an informed decision to terminate the pregnancy as this was an undesired pregnancy. She underwent an urgent right inferior gland parathyroidectomy. The mass was not fixed to underlying structures and was resected in-toto. Pathology showed chief cells with clear cytoplasm, and homogeneously round nuclei. There were no atypical mitoses, lymphovascular invasion, perineural invasion, or other invasive features concluding the final pathology as parathyroid adenoma. Discussion: Primary hyperparathyroidism predominantly affects postmenopausal women. Most common cause is parathyroid adenoma (80-85%). Parathyroid carcinoma is a rare cause of primary hyperparathyroidism (1-2%).80% people with parathyroid adenoma and primary hyperparathyroidism are diagnosed incidentally with mildly elevated calcium. PTH elevation is around 1.5-2 times the upper limit of normal. Symptoms are mild sequelae of hypercalcemia. PTH levels 5-10 times upper limit of normal, severe hypercalcemia (Ca > 14 mg/dl) or a hypercalcemia crisis are highly suspicious for parathyroid cancer. Surgical resection is the mainstay of treatment. Clinical criteria play a key role in early identification of parathyroid carcinoma in order to get these patients to surgery before tumor spread and improve long term survival. Our patient with significantly elevated calcium and parathyroid hormone had an atypical presentation of a parathyroid adenoma thus uncovering the limitations of clinical criteria in identifying a parathyroid carcinoma. Most of the patients with a parathyroid adenoma will eventually require surgery, however the technique, extent and urgency of surgery is affected based on clinical situations like these. The timing of the surgery could be detrimental when coupled with a scenario like our patient’s pregnancy wherein you could delay the surgery until the second trimester had the patient wanted to continue the pregnancy. This case highlights the importance of awareness of atypical presentations of parathyroid adenomas and to have a cautious approach in patients with clinical concern for parathyroid carcinoma especially if there is a potential benefit to delay surgery. Presentation: Saturday, June 17, 2023

Analysis of the morphology of retinal vascular cells in zebrafish (Danio rerio).

Background: Zebrafish (Danio rerio) have been established in recent years as a model organism to study Diabetic Retinopathy (DR). Loss of endothelial cells and pericytes is an early hallmark sign of developing DR in the mammalian retina. However, morphology, numbers, ratios, and distributions of different vascular cells in the retinal compartment in zebrafish have not yet been analyzed and compared with the mammalian retina. Methods: The retinal trypsin digest protocol was established on the zebrafish retina. Cell types were identified using the Tg(nflk:EGFP)-reporter line. Cells were quantified using quantitative morphometry. Results: Vascular cells in the zebrafish retina have distinct morphologies and locations. Nuclei of vascular mural cells appear as long and flat nuclei located near the vessel wall. Round nuclei within the vessel walls can be identified as endothelial cells. The vessel diameter decreases from central to peripheral parts of the retina. Additionally, the numbers of vascular cells decrease from central to peripheral parts of the retina. Discussion: The retinal trypsin digest protocol, which can be applied to the zebrafish retina, provides novel insights into the zebrafish retinal vascular architecture. Quantification of the different cell types shows that, in comparison to the mammalian retina, zebrafish have higher numbers of mural cells and an increased mural cell to endothelial cell ratio. This protocol enables to quantify mural cell and endothelial cell numbers, is easily adaptable to different transgenic and mutant zebrafish lines and will enable investigators to compare novel models on a single cell level.

Clear Cell Squamous Cell Carcinoma of the Maxillary Gingiva Associated with PIK3CA and HRAS Mutations: Report of a Case and Literature Review

BackgroundSquamous cell carcinoma (SCC) is the most common oral malignancy, and somatic mutations in some driver genes have been implicated in SCC development. Clear cell SCC (CCSCC) is a rare histological variant of SCC, and various clear cell neoplasms must be considered in the differential diagnosis of CCSCC in the oral cavity. Based on a limited number of CCSCC cases reported in the oral cavity, CCSCC is considered an aggressive variant of SCC with a poor prognosis; however, its genetic characteristics remain unknown.MethodsA maxillary gingival tumor in an 89-year-old female was described and investigated using immunohistochemical staining, special staining, fluorescence in situ hybridization, and next-generation sequencing (NGS) with a custom panel of driver genes, including those associated with SCC and clear cell neoplasm development.ResultsHistopathological examination revealed a proliferation of atypical epithelial cells with abundant clear cytoplasm and enlarged and centrally placed round nuclei. The tumor was exophytic with deep, penetrating proliferation. The atypical clear cells were continuous with the conventional SCC cells. Immunohistochemical analysis showed that the clear cells were positive for CK AE1/AE3 and CK5/6 and nuclear-positive for p63. In contrast, the clear cells were negative for αSMA, S100, HMB45, Melan-A, CD10, and p16. p53 immunoreactivity exhibited a wild-type expression pattern. Additionally, the clear cells were positive for periodic acid-Schiff (PAS) and negative for diastase-PAS, mucicarmine, and Alcian blue. Based on these results, the diagnosis of CCSCC was confirmed. Molecular analysis of the clear cells identified PIK3CA p.E542K (c.1624G>A) and HRAS p.G12A (c.35 G>C) somatic mutations classified as oncogenic. No pathogenic variants were identified in TP53, EWSR1, AKT1, PTEN, BRAF, KRAS, NRAS, RASA1, or MAML2.ConclusionsWe report a case of CCSCC of the oral cavity with PIK3CA and HRAS mutations. The identification of PIK3CA and/or HRAS mutations is rare in SCC; however, both mutations are important potential targets for antitumor therapy. A detailed analysis of gene mutations in CCSCC may lead to a better understanding of its biological behavior and an improved prognosis, as well as a differential diagnosis from other clear cell neoplasms.

Silk protein sericin: a promising therapy for Achilles tendinopathy-evidence from an experimental rat model.

This study investigated the efficacy of sericin in treating experimental Achilles tendinopathy (AT) in rats via the transforming growth factor-beta (TGF-β)/mothers against decapentaplegic (Smad) pathway compared with diclofenac sodium (DS). An AT model was induced in rats using collagenase enzyme type I and divided into 5 groups: C (control), AT (diseased control), ATS (AT treated with sericin), ATN (AT treated with DS), and ATSN (AT treated with sericin and DS). Sericin injection was given on the 3rd and 6th days by intratendinous injection (0.8 g/kg/mL), and DS was administered for 14 days by oral gavage (1.1 mg/kg/day). Serum concentrations of total oxidant-antioxidant status (TOS-TAS), TGF-β1, decorin, Smad2, and connective tissue growth factor (CTGF) were measured. Histopathologic and immunohistochemical (IHC) studies were conducted on Achilles tendon samples. The TOS, oxidative stress index (OSI), TGF-β1, Smad2, CTGF, and decorin serum concentrationswere significantly higher in AT than in C and significantly lower in ATS than in AT (P<0.05). Histopathological examination revealed that irregular fibers, degeneration, and round cell nuclei were significantly elevated in AT. Spindle-shaped fibers were similar to those in C, and degeneration was reduced in ATS. TGF-β1 and Smad2/3 expression was increased, and collagen type I alpha-1 (Col1A1) expression was decreased in AT vs. C (P=0.001). In the ATS, TGF-β1 and Smad2/3 expression decreased, and Col1A1 expression increased. The Bonar score significantly increased in the AT group (P =0.001) and significantly decreased in the ATS group (P =0.027). Sericin shows potential efficacy in reducing oxidative stress and modulating the TGF-β/Smad pathway in experimental AT models in rats. It may be a promising therapeutic agent for AT, warranting further clinical studies for validation. Key Points • This study revealed that sericin mitigates AT-induced damagethrough the TGF-β/Smad pathway in an AT rat model. • ELISA and IHC investigations corroborated the effectiveness ofsericin via the pivotal TGF-β/Smad pathway in tissue repair. • Evidence indicates that sericin enhances collagen synthesis,shapes tendon fiber structure, and diminishes histopathologicaldegeneration. • Sericin's antioxidant properties were reaffirmed in its AT treatmentapplication.

Histological and morphometric analysis of skeletal muscle in some vertebrates

The skeletal system is primarily driven by the skeletal muscles to produce kinematic movements. The study evaluates the histological and morphometric properties of skeletal muscle in Clarias gariepenus (Cl. gariepinus), Bufo bufo (B. bufo), Agama agama (A. agama), Columba livia domestica (C. domestica) and Rattus rattus (R. rattus). The study was carried in order to relate the similarities and differences of skeletal muscles in these species with evolutionary trend. The epaxial muscle of Cl. Gariepinus, the biceps femoris muscle of B. bufo, R. rattus, puboischiotibialis of A. agama, and pectoral muscle from C. domestica were removed and assessed grossly for physical appearance then processed for histological analysis. The diameters of the muscle fibers were measured and one-way analysis of variance was used to compare the differences. The muscles of Cl. gariepinus, B. bufo and A. agama appeared whitish with scanty fusiform nucleus and large intermuscular space. However, the muscles of C. domestica and R. rattus appeared red with distinct round nucleus and small intermuscular space. No significant difference (P&gt;0.05) was observed in the muscle diameter of Cl. gariepinus (8.86±0.13µm) compared to B. bufo (8.25±0.27µm). The muscle diameter of A. agama (10.18±0.25µm) was significantly higher (P&lt;0.05) relative to Cl. gariepinus (8.86±0.13µm), B. bufo (8.25±0.27µm), C. domestica (3.38±0.13µm) and R. rattus (4.66±0.15µm). Conclusively, non-tetrapod vertebrates (Cl. gariepinus, B. bufo, and A. agama) have simple, white-colored skeletal muscle with few flat-shaped nuclei and large fiber diameters while higher vertebrates (C. domestica and R. rattus) have complex, red-colored skeletal muscle with numerous oval-shaped nucleus and small fiber diameter.

Heterotopic ovarian hilus cells of the salpinx: a case report and literature review

Ovarian hilus cells (OHCs), a counterpart of testicular Leydig cells, are usually found in the ovarian poles and produce androstenedione. Their origin remains a matter of debate, although OHCs are assumed to come from the adrenogenital primordium. OHCs are rarely observed around the poles of the ovary, including the mesoovarium, stroma (perisalpinx) of the salpinx, and the wall of paratubal cysts. Their clinical and pathological characteristics are not well-known because of their rarity. Herein, we present a case of ectopic OHCs in a 48-year-old woman. The patient underwent total hysterectomy and bilateral salpingectomy for vaginal bleeding due to multiple leiomyomas. We incidentally found OHCs in the stroma of the infundibulum of the salpinx, just beneath the tubal epithelium. Their size was less than 1 mm, and they were composed of large cells with central round nuclei and abundant clear or granular cytoplasm. OHCs share morphological and immunohistochemical profiles with ectopic adrenal glands, and the differential diagnosis is sometimes difficult. They do not exhibit microscopic encapsulation or the normal adrenal cortex zonation pattern. The patient was discharged and did not show any abnormal findings during 19 months of follow-up. Analyzing the characteristics of testicular Leydig cells will help understand how OHCs develop and why heterotopic OHCs occur in and around the salpinges.

Pinealocytes and Glia Cells in the Pineal Gland of the African Straw-Coloured Fruit Bat (Eidolon helvum)

Abstract The mammalian pineal gland is a structure that in recent years has been extensively studied, due to its functions and the hormones it produces. Bats are the only known flying mammals, with the order having a large number of species. This study was carried out to investigate the gross and histological features of the pineal gland of the Eidolon helvum, the African fruit bat, using male and female subjects. Eight free flying E. helvum (4 males, 4 females) were captured using mist nets. The pineal gland was grossly observed to be very small in size, oval in shape, and covered by the pia mater. Histological examination revealed two populations of pinealocytes – Types I and II – oval structures possessing acidophilic cytoplasm and large, round nuclei. Pinealocytes were seen to appear singly or in clusters, having no particular arrangement. Sexual dimorphism was observed, with the females having less density in population of pinealocytes at the peripheral region of the gland. This was consistent in all subjects examined. Astrocytic appearance was typical with long and slender processes, and perivascular and capsular microglia were observed. The glia cells were observed to be abundant in the parenchyma and around the capsule. There was no sensitivity to NeuN antibody. Results obtained may find application in behavioural and comparative neuroscience.

Postnatal Development of the Amygdala of Rat Offspring after Prenatal Exposure to Valproic Acid: A Histological and Morphometric Study

Abstract Background The amygdala has a major role in emotion regulation, social behavior development and reward learning. Being subject to several factors makes it vulnerable to numerous neurodevelopmental disorders. Aim of the Work The aim of the current work was to study the structural and morphometric changes of the amygdala of rat's offspring following maternal exposure to valproic acid during pregnancy and to compare it with the control. Material and methods 20 adult female and 10 adult male Sprague dawley albino rats weighing 150-200 g body weight were used in the current study. After induction of pregnancy, pregnant rats were divided into two groups 6 in each group; control group (I) and valproic acid treated group (II). Pregnant rats received one ml of saline in group (I) or 500 mg/kg BW valproic acid dissolved in one ml saline in group (II) by oral gavage on day 11-13 of pregnancy. Pregnant rats were observed till end of pregnancy and four male newborn rats were sacrificed at each time point of the experiment from the two groups (week1, 2, &amp;3). Brains were extracted and processed for paraffin blocks and sectioned then stained with H&amp;E, cresyl violet stains and immunohistochemically stained for GFAP antibody. Image analysis &amp;morphometry were performed on stained sections for; number of neurons, number of oligodendrocytes, surface area of basal nucleus of amygdala and density of GFAP staining. Results On examination of H&amp;E-stained sections of control groups during the first three weeks postnatally, the amygdala contained mainly three types of cells: pyramidal cells, oligodendrocytes, and astrocytes. Pyramidal cells had basophilic cytoplasm with large rounded open face nuclei and prominent nucleoli. Some pyramidal cells showed apical dendrites. The oligodendrocytes were seen to have rounded nuclei with condensed chromatin surrounded by thin cytoplasmic ring. Astrocytes possessed nuclei with pale vesicular chromatin pattern less dense than that of oligodendrocytes and showed relatively small indistinct nucleoli. Morphometric studies using image analysis had recorded statistically non-significant changes in the number of neurons during development, however concerning oligodendrocytes there was a statistically significant increase in their number in week 2 and 3 when compared to week 1. Using Nissl staining (cresyl violet) it demonstrated negative or very weak staining in the first week postnatally, the staining becomes more obvious in the next 2 weeks postnatally. GFAP immune stained sections of the amygdala of the 3 weeks revealed mild positive immune reaction. There was no statistically significant difference regarding density of staining across the three weeks. The surface area of the basal nucleus of amygdala of the control groups showed statistically significant increase during development across the three weeks. On examination of H&amp;E-stained sections of VPA treated groups during the first three weeks postnatally, numerous cells with pyknotic deeply basophilic stained nuclei were seen. Morphometric studies on the same groups using image analysis recorded statistically significant decrease in neurons and oligodendrocytes number when compared to control. On examination of the cresyl violet stained sections, there were numerous darkly stained degenerating cells that did not reveal the presence of Nissl granules denoting chromatolysis. On examining GFAP immune stained sections of the amygdala, week 2 and 3 postnatally showed strong positive immune reaction. The surface area of the basal nucleus of amygdala of VPA treated groups of weeks 1 showed a highly significant decrease when compared to week 1 of control groups, however the surface area of the amygdala of week 2 and 3 showed an increase more than the increase recorded in the control groups. The increase in the surface area was non-significant in week 2 but highly significant in week 3. Conclusion Results of the present experimental study demonstrated the developmental structural changes of the rat amygdala across the first three weeks of postnatal life. The study also explored the devastating effect of the prenatal exposure to VPA on the structure of the amygdala leading to neuronal death and gliosis particularly in the third week of age postnatally.

Electron microscopic changes in fibroblastic sarcoid in horses

The article presents the results of ultrastructural research on fibroblastic sarcoid, which is one of the most aggressive types of sarcoid in horses. A retrospective study on the prevalence of sarcoids in horses in Ukraine’s western regions was conducted in 2019–2023. It was found that during the period of 2022–2023, the number of horses with sarcoids increased sharply by 15.6% compared to the period 2020–2021. This was associated with the mass movement of animals from eastern regions due to the onset of the war in Ukraine. The fibroblastic type of sarcoid was predominantly localized in the abdominal wall and groin area. To study the ultrastructure of sarcoids, they were surgically removed by excising undamaged skin portions under general anesthesia, adhering to all requirements of the European Convention for the Protection of Animals used for Experimental and Other Scientific Purposes. Electronograms revealed that the tumour formation mainly consisted of fibroblasts of varying differentiation degrees: significant number of vesicles, markedly expanded endoplasmic reticulum cisterns, and irregularly shaped nuclei with numerous invaginations. Open nuclear pores were observed in most nuclei. Active formation of a large number of capillaries was noted between the collagen matrix, fibroblasts, and fibrocytes, indicated by the increased number of endotheliocytes with pseudopodia on the cytoplasm’s marginal part and the basal surface. This suggests their embryonic type. Endotheliocytes contained large round nuclei and a significant number of mitochondria in the cytoplasm. There were both bright and dark endotheliocytes in the blood capillaries’ venous section. The cytoplasm of bright endotheliocytes contained myofilaments, individual mitochondria, free ribosomes, peroxisomes, and vesicles of various sizes. The plasmalemma formed small pseudopodia. The capillary lumen showed moderate electron density. Additionally, high platelet activity was observed, manifested by cell adhesion to the marginal part of the endotheliocyte cytoplasm. It should be noted that most capillaries were of the venous type, as indicated by the endotheliocytes’ height, the presence of an increased number of mitochondria and vesicles. The entire cellular pool was localized among the massive framework consisting of collagen fibers. Elastic fibers could not be detected in the fibrillar component.

Typical carcinoid in right middle lobe of pulmonary hypoplasia

BackgroundPulmonary typical carcinoid occurring in hypoplasia of the right middle lobe is very rare.Case presentationA routine examination's chest X-ray revealed an abnormal shadow in the right middle lung field of an 82-year-old Japanese woman. A chest computed tomography scan showed a solid 2.5 × 2.0-cm nodule in the very small right middle lobe. A trans-bronchial lung biopsy of the mass in the right middle lobe was performed; it revealed atypical cells with round nuclei growing in multiple foci, and immunostaining was positive for chromogranin A, synaptophysin and CD56, suggesting pulmonary carcinoid. The preoperative clinical diagnosis of primary lung cancer, cT1cN0M0 stage IA3 was considered. A right middle lobectomy and mediastinal lymph node dissection were performed by video-assisted thoracic surgery. Intraoperatively, the middle lobe of the right lung was very small, with 1- to 2-mm-dia. pulmonary arteries and veins that were considered hypoplastic. The final histopathological diagnosis was typical carcinoid, pT2aN0M0 stage IB based on the presence of pleural invasion.ConclusionsIncluding the present patient, only nine cases of lung cancer occurring within pulmonary hypoplasia have been reported, most of which were typical carcinoid.

Adamantinoma-like Ewing Sarcoma (ALES) May Harbor FUS Rearrangements : A Potential Diagnostic Pitfall.

Adamantinoma-like Ewing sarcoma (ALES) is a rare malignancy currently considered a variant of Ewing sarcoma with most known cases harboring EWSR1 rearrangements. Herein we present a series of 6 cases of EWSR1 -negative ALES. The tumors arose in the sinonasal tract (n=3), major salivary glands (submandibular gland=1; parotid=1), and anterior mediastinum (n=1) in patients ranging from 25 to 79 years of age. Most tumors were basaloid in appearance, growing in large nests separated by interlobular fibrosis without overt squamous pearls. However, 1 case closely resembled a well-differentiated neuroendocrine tumor with uniformly round nuclei, eosinophilic cytoplasm, and trabecular architecture. All cases were diffusely positive for pan-cytokeratin, p40 or p63, and CD99. A subset of cases showed diffuse reactivity for synaptophysin, including 1 sinonasal tumor which also demonstrated sustentacular S100 protein expression. Molecular testing showed FUS rearrangements in all cases. Gene partners included known ETS family members FEV (n=2) and FLI1 (n=1). Our results expand the molecular diagnostic considerations for ALES to include FUS rearrangements. We also show that ALES may harbor FUS :: FLI1 fusion, which has not been previously reported in the Ewing family of tumors. Furthermore, ALES may show unusual histologic and immunophenotypic features that can overlap with olfactory carcinoma including S100-positive sustentacular cells. ALES should be considered in the diagnostic differential of small round cell tumors and tumors with neuroendocrine differentiation with immunohistochemical workup to include p40 and CD99/NKX2.2.

Dermoscopy of nodular hydradenoma

Hidradenoma Nodularis (HN) is a benign adnexal tumor that was generally considered to be of eccrine origin, although apocrine forms also exist. It develops from sweat glands located in the superficial or deep dermis or sometimes even in the subcutaneous tissue. Clinically, it appears as a small, solitary, firm papule, which may be dark or transparent in color, although usually a single papule, it is less common to find multiple papules [1]. the confirmation of the diagnosis is based on the histology spectra which will find a dermal nodule well bounded by a pseudocapsule of collagen which lobules filled with epithelial cells and tubular structures. Tumor proliferation in HN is very typical, dense and often hyalinized, making it an important element in the diagnosis. It is characterized by two cell types: polyhedral cells with elongated nuclei and slightly basophilic cytoplasm, and cells with small rounded nuclei and clear cytoplasm [2]. few publications in the literature have described the dermoscopic aspect which has structureless white-pink areas; red lacunalike areas; and hairpin, dotted, and linear irregular vessels [3,4], we describe a new case of dermoscopy of nodular hydradenoma.

Effects of Soya Beans and Brown Beans in Degenerative Changes Expressed in Cortical Neun Immunoreactivity in a Casava-Induced Konzo Disease Rat Model

Background: Konzo is related to a habit of long-term consumption of cassava; it is a chronic disease with quite different symptoms from acute cassava poisoning. The ameliorative effect of soya beans and brown beans in degenerative changes expressed in cortical NeuN immunoreactivity in a konzo disease rat model was investigated in the present study. Materials and Methods: 30 male wistar rats weighing 200-250g were assigned to Group 1 (Control, n=5) and was fed on animal pellet, whereas Group 2 (Protein, n=5) was administered with protein food (Soya beans+Brown bean). Bitter cassava flour was provided to Group 3 (cassava-induced Konzo, n=15). Protein (Soya beans+Brown bean) and bitter cassava flour were administered to Group 4 (protein treatment group, n=5). The Cerebrum (primary motor cortex) region was harvested through transcardiac perfusion for histological and immunohistochemical staining. Image J was used to quantify the neurons in the motor cortex. We examined general structures in the primary motor cortex with hematoxylin and eosin (H&amp;E) stain. Result and Discussion: H&amp;E demonstration showed distinct pyramidal neurons with large round nucleus, and prominent nucleoli in the Control group. Interspersed between the pyramidal neurons were numerous glia cells. Protein (Soya beans+Brown bean) group, Cassava group, and Cassava + Protein group also showed intact histology of the cortex compared to control usig H&amp;E stain. NeuN stains analyzed immunoreactive neurons. The rats fed on cassava showed significant decrease in the NeuN immunoreactive neurons at ***[p&lt;.001] compared to the Control and Protein administered group, whereas Cassava + Protein groups showed decrease NeuN immunoreactive neurons at**p&lt;0.1 compared to Protein group(Soya beans+Brown bean). Conclusions: This study has provided an empirical data on the ameliorative effect of soya beans and brown beans supplement on Cassava-induced konzo disease animal model. This data will be very useful to Anatomists, konzo researchers and neuroscientist.

Palisading Adenocarcinoma: A Morphologically Unique Salivary Gland Tumor With a Neuroendocrine-like Appearance and a Predilection for the Sublingual Glands of Women.

Adenocarcinoma, not otherwise specified (NOS) is a heterogenous group of salivary gland tumors that likely contains distinct tumors that have not yet been characterized. Indeed, in recent years, cases previously diagnosed as adenocarcinoma, NOS have been recategorized into novel tumor designations such as secretory carcinoma, microsecretory adenocarcinoma, and sclerosing microcystic adenocarcinoma. We sought to describe a distinctive, hitherto-undescribed salivary gland tumor encountered in the authors' practices. Cases were pulled from the surgical pathology archives of the authors' institutions. Histologic, immunohistochemical, and clinical findings were tabulated, and targeted next-generation sequencing was performed on all cases. Nine cases were identified, arising in 8 women and 1 man ranging from 45 to 74 years (mean, 56.7y). Seven tumors (78%) arose in the sublingual gland, while 2 (22%) arose in the submandibular gland. The cases shared a distinctive morphologic appearance. They were biphasic, with ducts scattered among a predominant polygonal cell with round nuclei, prominent nucleoli, and pale eosinophilic cytoplasm. These cells were arranged as trabeculae and palisaded as pseudorosettes around hyalinized stroma and vessels, resembling a neuroendocrine tumor. Four of the cases were well-circumscribed, while the remaining 5 showed infiltrative growth including perineural invasion in 2 (22%) and lymphovascular invasion in 1 (11%). Mitotic rates were low (mean, 2.2/10HPFs); necrosis was absent. By immunohistochemistry, the predominant cell type was strongly positive for CD56 (9 of 9) and variably positive for pan-cytokeratin (AE1/AE3) (7 of 9) with patchy S100 (4 of 9), but negative for synaptophysin (0 of 9) and chromogranin (0 of 9), while the ducts were strongly positive for pan-cytokeratin (AE1/AE3) (9 of 9) and CK5/6 (7 of 7). Next-generation sequencing did not reveal any fusions or obvious driver mutations. All cases were resected surgically, with external beam radiation also done in 1 case. Follow-up was available in 8 cases; there were no metastases or recurrences after 4 to 160 months (mean, 53.1mo). A dual population of scattered ducts with a predominance of CD56-positive neuroendocrine-like cells characterizes a unique salivary gland tumor which is often encountered in the sublingual glands of women, for which we propose the term "palisading adenocarcinoma." Although the tumor was biphasic and had a neuroendocrine-like appearance, it lacked convincing immunohistochemical evidence of myoepithelial or neuroendocrine differentiation. Although a subset showed unequivocally invasive growth, this tumor appears to behave in an indolent manner. Moving forward, recognition of palisading adenocarcinoma and its separation from other salivary adenocarcinomas, NOS will facilitate a better understanding of the characteristics of this previously unrecognized tumor.

Hepatocellular carcinoma presenting as a chest wall mass - An unusual case diagnosed on cytology with review of literature.

Hepatocellular carcinoma (HCC) is associated with an aggressive behavior and a strong tendency for extrahepatic metastasis. Although 5%-15% patients have metastases at diagnosis, presentation with symptoms exclusively related to extrahepatic metastases is rare. An 82-year-old male presented with an isolated left anterolateral chest wall swelling. Ultrasonography revealed a soft tissue mass involving the anterior chest wall with adjacent rib erosion. Serum protein electrophoresis showed increase in beta-2 region. A clinical diagnosis of multiple myeloma was considered. Fine needle aspiration cytology from the swelling showed loosely cohesive clusters of polygonal cells with traversing blood vessels. Cells showed abundant vacuolated to granular cytoplasm, round nuclei with frequent intranuclear cytoplasmic inclusions. A differential of metastatic HCC and renal cell carcinoma was considered. Subsequent imaging showed a 12 cm mass in the liver. Biopsy from chest wall mass with immunohistochemistry confirmed the diagnosis. Lungs and lymph nodes are the commonest sites for metastatic HCC; presentation as chest wall metastasis is rarely reported. The classical cytomorphology of HCC proved useful in diagnosing metastasis at a rare site. Recent studies have shown that beta-2-globulin is a promising biomarker for early diagnosis of HCC in patients with chronic liver disease.