Cytoplasmic Ratio Research Articles

Salivary Gland Adaptation to Dietary Inclusion of Hydrolysable Tannins in Boars.

Simple SummaryTannins have traditionally been avoided in animal nutrition due to their anti-nutritive effects. However, recent studies reported hydrolysable tannins as beneficial additives that have antimutagenic, anticarcinogenic, antidiarrheal, and antiulcerogenic effects on animals. In a study testing the inclusion of hydrolysable tannins as a potential nutritive factor to reduce boar taint in entire males, significant enlargement of the parotid glands (parotidomegaly) was observed. In this study, we aimed to determine the morphological and immunohistochemical basis for the observed parotidomegaly. We discovered that enlargement of glandular lobules and acinar area, an increased ratio between the nucleus and cytoplasm of serous cells, and increased excretion of proline-rich proteins (PRPs) were characteristic of the experimental group that received the highest dietary tannin intake. The mandibular salivary gland, on the other hand, did not show significant morphological changes among the experimental groups. This suggests increased functional activity of the parotid salivary glands as the first and most important line of defense against high dietary tannin and its potential negative effects.The ingestion of hydrolysable tannins as a potential nutrient to reduce boar odor in entire males results in the significant enlargement of parotid glands (parotidomegaly). The objective of this study was to characterize the effects of different levels of hydrolysable tannins in the diet of fattening boars (n = 24) on salivary gland morphology and proline-rich protein (PRP) expression at the histological level. Four treatment groups of pigs (n = 6 per group) were fed either a control (T0) or experimental diet, where the T0 diet was supplemented with 1% (T1), 2% (T2), or 3% (T3) of the hydrolysable tannin-rich extract Farmatan®. After slaughter, the parotid and mandibular glands of the experimental pigs were harvested and dissected for staining using Goldner’s Trichrome method, and immunohistochemical studies with antibodies against PRPs. Morphometric analysis was performed on microtome sections of both salivary glands, to measure the acinar area, the lobular area, the area of the secretory ductal cells, and the sizes of glandular cells and their nuclei. Histological assessment revealed that significant parotidomegaly was only present in the T3 group, based on the presence of larger glandular lobules, acinar areas, and their higher nucleus to cytoplasm ratio. The immunohistochemical method, supported by color intensity measurements, indicated significant increases in basic PRPs (PRB2) in the T3 and acidic PRPs (PRH1/2) in the T1 groups. Tannin supplementation did not affect the histo-morphological properties of the mandibular gland. This study confirms that pigs can adapt to a tannin-rich diet by making structural changes in their parotid salivary gland, indicating its higher functional activity.

The morphogenesis of the rabbit meibomian gland in relation to sex hormones: Immunohistochemical and transmission electron microscopy studies

Rabbits have been proposed as a model for the human meibomian gland (MG), a large specific sebaceous gland in the eyelid that consists of secretory acini arranged laterally and related to the central duct via short ductules, with the central duct continuing as an excretory duct to open at the free margin of the lid. First detected at embryonic day 18 as an aggregation of mesenchymal cells in the tarsal plate, it completes its development approximately 2 weeks postnatal when the separation of the eyelids is completed. The Transmission electron microscopy supports the meibocytes’ gradient maturation to the meibum’s synthesis. While the differentiating cells, their cytoplasm, are well packed with lipid droplets, the basal cells are characterized by a high nuclear to cytoplasm ratio. The androgen and estrogen receptor proteins are expressed in the basal cell and the meibocytes, and increase in age increases in the expression of these proteins. Additionally, the cytokeratin (CK14) is expressed in the basal and differentiating cells of the acini and the ductal epithelium. Therefore, the duct cells of the MG are common in all stem cells. These data concluded that the MG plays a major role in maintaining the health of the ocular surface and preservation of visual acuity. Any abnormalities in the structure of the MG lead to its dysfunction and changes in lipid secretion.

Isolation of Swine Bone Marrow Lin-/CD45-/CD133 + Cells and Cardio-protective Effects of its Exosomes

BackgroundThe identification in murine bone marrow (BM) of CD133 + /Lin-/CD45- cells, possessing several features of pluripotent stem cells, encouraged us to investigate if similar population of cells could be also isolated from the swine BM. Heart failure is the terminal stage of many cardiovascular diseases, and its key pathological basis is cardiac fibrosis (CF). Research showed that stem cell derived exosomes may play a critical role in cardiac fibrosis. The effect of exosomes (Exos) on CF has remained unclear.ObjectiveTo establish an isolation and amplification method of CD133 + /Lin-/CD45- cells from newbron swine BM in vitro, explore an highly efficient method to enrich swine bone marrow derived CD133 + /Lin-/CD45- cells and probe into their biological characteristics further. Furher more, to extract exosomes from it and explore its effect on CF.MethodsThe mononuclear cells isolated from swine bone marrow by red blood cell (RBC) lysing buffer were coated by adding FcR blocking solution and coupled with CD133 antibody immunomagnetic beads, obtaining CD133 + cell group via Magnetic Activated Cell Sorting (MACS). In steps, the CD133 + /Lin-/CD45- cells were collected by fluorescence-activated cell sorting (FACS) labeled with CD133, Lin and CD45 antibodies, which were cultured and amplified in vitro. The biological features of CD133 + /Lin-/CD45- cells were studied in different aspects, including morphological trait observed with inverted microscope, ultrastructural characteristics observed under transmission electron microscope, expression of pluripotent markersidentified by immunofluorescent staining and Alkaline phosphatase staining.The Exos were extracted using a sequential centrifugation approach and its effects on CF were analyzed in Angiotensin II (Ang-II) induced-cardiac fibrosis in vivo. Rats in each group were treated for 4 weeks, and 2D echocardiography was adopted to evaluate the heart function. The degree of cardiac fibrosis was assessed by Hematoxylin–Eosin (HE) and Masson's trichrome staining.ResultsThe CD133 + /Lin-/CD45- cells accounted for about 0.2%-0.5% of the total mononuclear cells isolated from swine bone marrow. The combination of MACS and FACS to extract CD133 + /Lin-/CD45- cells could improved efficiency and reduced cell apoptosis. The CD133 + /Lin-/CD45- cells featured typical traits of pluripotent stem cells, the nucleus is large, mainly composed of euchromatin, with less cytoplasm and larger nucleoplasmic ratio, which expressed pluripotent markers (SSEA-1, Oct-4, Nanog and Sox-2) and alkaline phosphatase staining was positive.Animal experiment indicated that the cardiac injury related indexes (BNP、cTnI、CK-MB and TNF-α), the expression of key gene Smad3 and the degree of cardiac fibrosis in Exo treatment group were significantly reduced compared with the control group. 4 weeks after the treatment, cardiac ejection fraction (EF) value in the model group showed a remarkable decrease, indicating the induction of HF model. While Exo elevated the EF values, demonstrating cardio-protective effects.ConclusionThe CD133 + /Lin-/CD45- cells derived from swine bone marrow were successfully isolated and amplified, laying a good foundation for further research on this promising therapeutic cell. The Exos may be a promising potential treatment strategy for CF.Graphical

Spontaneous tumor lysis syndrome in hyperaggressive mantle cell lymphoma.

An otherwise well 61-year-old male presented to his primary care practitioner with drenching sweats, weight loss, massive splenomegaly (30 cm as measured by computed tomography [CT], Figure 1A) and a lymphocytosis (white blood cell [WBC] at diagnosis 150 × 109/L). The peripheral smear was notable for the presence of pleomorphic abnormal lymphocytes, with clumped chromatin, high nuclear:cytoplasm ratio and irregular nuclei (Figure 1B). Peripheral blood flow cytometry revealed a kappa light chain restricted population of mature B-cells which co-expressed CD20 and CD5 (Figure 1C). CD23 and CD10 were negative. Fluorescent in situ hybridization revealed a CCND1/IGH rearrangement, consistent with a diagnosis of advanced stage mantle cell lymphoma (MCL). Approximately 3 weeks after diagnosis, he was admitted electively for chemotherapy. On admission to the unit he was well, with normal vital signs and a normal baseline tumor lysis profile (K, Phos, Ca, urate and creatinine all within institution limits). His WBC was noted to have increased modestly to 264 × 109/L. Unfortunately, over the course of the next 12 h, prior to the administration of any systemic anti–cancer therapy (including steroids), he developed spontaneous tumor lysis syndrome and died of catastrophic metabolic derangement, despite aggressive supportive care. He developed severe, refractory hyperkalemia (7.5 mmol/L), severe hyperphosphatemia (3.83 mmol/L) and profound metabolic acidosis (pH <6.9, HCO3– 3 mmol/L, lactate 17 mmol/L). His WBC had also climbed explosively to 695 × 109/L (Figure 1D), putting him at significant risk of leukostasis. I postulate that, via clonal evolution, his disease acquired a hyperproliferative phenotype, resulting in fulminant, spontaneous tumor lysis syndrome. There are limited reports of secondary translocations involving the MYC proto-oncogene in patients with MCL. This is associated with aggressive clinical course and short overall survival; however there are no reports of MCL behaving this aggressively in the literature. Practitioners should be aware of this rare but potentially devastating complication of MCL. James Wilson wrote the manuscript and took pictures. The author declares they have no conflicts of interest. The author received no specific funding for this work. No research on human was performed on this study. Data sharing not applicable—no new data generated.

Basaloid Squamous Cell Carcinoma of the Uterine Cervix in a Northern Nigerian Hospital: A Case Report and Review of Literature

Basaloid squamous cell carcinoma occurs very rarely in the uterine cervix. Basaloid squamous cell carcinoma is a form of squamous cell carcinoma with a poor prognosis that primarily affects persons in their late sixties and early seventies. This paper describes a case of Basaloid squamous cell carcinoma in the fifth decade that had spread to the myometrium, ovaries, and fallopian tubes. The macroscopic findings of a total abdominal hysterectomy and bilateral salphingo-oophorectomy revealed an ulcerative fungating tumor distorting the whole cervix and macroscopic invasion of the posterior uterine wall. Histologic features revealed a Basaloid tumour disposed in nest exhibiting peripheral palisading and stromal retraction with most of the nest having comedo pattern of necrosis. The tumour cells are fairly small uniform round cells, with scanty cytoplasm having increased nuclei cytoplasmic ratio with vesicular to open nuclei. Immunohistochemical stains; Cytokeratins, EMA, p63 and Ki-67 showed cytokeratin positivity and EMA strong and diffuse positivity. Keywords: Basaloid Squamous cell Carcinoma, uterine cervix, p63, EMA, Ber-EP4

Histologic Growth Patterns in Clear Cell Renal Cell Carcinoma Stratify Patients into Survival Risk Groups.

Genomic and morphologic heterogeneity in clear cell renal cell carcinoma (ccRCC) presents a barrier to prognostication and treatment decisions. Data from pathology are used with clinical markers to predict disease progression after nephrectomy. However, determining the risk of cancer recurrence, and survival with metastatic cancer remains challenging. Recently, analysis of histologic growth patterns (HGP) in ccRCC revealed promising associations with survival outcomes. To investigate whether HGPs can be used to predict overall survival (OS) after nephrectomy, we examined 24 HGPs in primary tumors of 147 patients that included 107 patients with metastatic disease. The median number of HGPs per case was 5 and was greater in metastatic and larger tumors. After adjustment for 6 pathologic and demographic variables, HGPs were significantly associated with OS post nephrectomy. Small nests, expansile nests and nests with high nuclear to cytoplasmic ratio were associated with favorable outcomes; while spindled low grade, fused nests/solid sheets, rhabdoid, and sarcomatoid patterns were associated with unfavorable outcomes. A 3-tiered and a 2-tiered risk model were developed based on combinations of HGPs. The models performed equally well as WHO/ISUP nucleolar plus necrosis grade (necrosis grade), and better than WHO/ISUP nucleolar grade alone in predicting OS at the median OS of 6 years. Pairwise correlations between HGPs revealed 2 tumor evolutionary branches that differed in risk of metastatic disease: one with mesenchymal differentiation, and other with epithelial tubulopapillary differentiation. While 44 of 107 (41%) patients with metastatic ccRCC displayed evidence of mesenchymal differentiation, mesenchymal features were only observed in 1 of 40 (3%) patients without evidence of metastatic disease. These findings suggest that HGPs may provide a novel path to refine the estimation of OS after nephrectomy and to determine the molecular basis of tumor evolution.

Stress Relaxation and Composition of Hydrazone-Crosslinked Hybrid Biopolymer-Synthetic Hydrogels Determine Spreading and Secretory Properties of MSCs.

The extracellular matrix plays a critical role in mechanosensing and thereby influences the secretory properties of bone-marrow-derived mesenchymal stem/stromal cells (MSCs). As a result, interest has grown in the development of biomaterials with tunable properties for the expansion and delivery of MSCs that are used in cell-based therapies. Herein, stress-relaxing hydrogels are synthesized as hybrid networks containing both biopolymer and synthetic macromer components. Hyaluronic acid is functionalized with either aldehyde or hydrazide groups to form covalent adaptable hydrazone networks, which are stabilized by poly(ethylene glycol) functionalized with bicyclononyne and heterobifunctional small molecule crosslinkers containing azide and benzaldehyde moieties. Tuning the composition of these gels allows for controlled variation in the characteristic timescale for stress relaxation and the amount of stress relaxed. Over this compositional space, MSCs are observed to spread in formulations with higher degrees of adaptability, with aspect ratios of 1.60± 0.18, and YAP nuclear:cytoplasm ratios of 6.5± 1.3. Finally, a maximum MSC pericellular protein thickness of 1.45± 0.38µm occurred in highly stress-relaxing gels, compared to 1.05± 0.25µm in non-adaptable controls. Collectively, this study contributes a new understanding of the role of compositionally defined stress relaxation on MSCs mechanosensing and secretion.

Cytologic features of micropapillary variant urothelial carcinoma in urinary tract cytology: Case series and review of literature.

The micropapillary variant of urothelial carcinoma (MPVUC) is rare and aggressive. Surgical specimens often show atypical micro-clusters (AMCs) of cells with hyperchromatic, pyknotic, peripheral, irregular nuclei with variable nuclear to cytoplasmic ratios. We reviewed urinary tract cytology (UTC) from patients with MPVUC and hypothesized that AMCs would be present similar to those in surgical specimens. The archives were searched from 2000 to 2020 for patients with surgical cases with either MPVUC or conventional high-grade urothelial carcinoma (HGUC) and with prior abnormal UTC. Two pathologists reviewed UTC cases and controls in a blinded manner for AMCs, with quantitation of none, low, moderate, and high. Interrater reliability was compared by quadratic weighted Cohen's Kappa test. The association between numerical average score and MPVUC status was determined by logistic regression. Five patients with invasive MPVUC, one patient with a noninvasive micropapillary component, and 15 control patients with conventional HGUC were included. All patients had prior or concurrent abnormal UTC samples. Increasing category of quantities of AMCs on cytology was associated with micropapillary status (OR 7.9, 95% CI 2.7-118, p= .045), with moderate agreement between raters (Cohen's Kappa 0.54, 95% CI 0.19-0.89, p= .004). In patients with MPVUC on surgical specimen, AMCs were frequently observed on cytology. Similar atypical clusters were observed in patients with nonmicropapillary HGUC, albeit at lower frequency. However, given the WHO recommendation to diagnose micropapillary only if an invasive micropapillary component is present, a specific diagnosis of MPVUC on UTC cannot be based solely on the presence of AMCs.

A Presurgical-Window Intervention Trial of Isothiocyanate-Rich Broccoli Sprout Extract in Patients with Breast Cancer.

Dietary isothiocyanates (ITCs) from cruciferous vegetables have shown potent anti-breast cancer activities in preclinical models, but their anticancer effects in vivo in breast cancer patients remain elusive. A proof-of-principle, presurgical window of opportunity trial is conducted to assess the anticancer effects of dietary ITCs in breast cancer patients. Thirty postmenopausal breast cancer patients are randomly assigned to receive ITC-rich broccoli sprout extract (BSE) (200µmol ITC per day) or a placebo for 2 weeks. Expression of biomarkers related to ITCs functions are measured in breast cancer tissue specimens at pre- and post-interventions using immunohistochemistry staining. First morning urine samples are collected at both timepoints for proteomic analysis. Overall, the study shows high compliance (100%) and low toxicity (no grade 4 adverse event). Trends of increase in cleaved caspase 3 and tumor-infiltrating lymphocytes (TILs) and trends of decrease in Ki-67 and nuclear to cytoplasm ratio of estrogen receptor (ER)-α are observed in the BSE arm only, consistent with the significantly altered signaling pathways identified in urinary proteomic analysis. Anticancer activities of ITCs are observed in breast cancer patients, supporting the potential beneficial roles of ITC-containing cruciferous vegetables in breast cancer prognosis.

Abstract 191: Cops5-mediated Nuclear Exclusion Of P27 Is Responsible For The Promotion Of Vascular Smooth Muscle Cells Proliferation By Cops8 Hypomorphism

Background: The COP9 signalosome (CSN) holocomplex is formed by 8 stoichiometric protein subunits: COPS1 through COPS8. Despite the strong evidence linking CSN to cell cycle regulation, direct study of CSN in VSMC proliferation is lacking. Our prior data show that vascular injury significantly increases Cops8 proteins in wild-type mice and induces more severe neointimal thickening and increased VSMC proliferation in Cops8 hypomorphic (Cops8-hypo) mice compared to non-hypomorphic littermates (CTL), suggesting an important role of the CSN in vascular pathology. The present study was carried out to further elucidate the mechanism underlying this phenomenon. We hypothesize that increased COPS5-mediated nuclear exclusion of p27 contributes to the promotion of VSMC proliferation by COPS8 hypomorphism. Methods and Results: We isolated and cultured VSMCs from the COPS8-hypo and CTL mice and used the treatment with platelet-derived growth factor (PDGF-BB) to mimic vascular injury. In response to PDGF-BB, COPS8-hypo VSMCs displayed increased hyperproliferation compared to CTL VSMCs. We detected markedly smaller nuclear to cytoplasmic ratio for p27 and COPS5; and native gel electrophoresis revealed increases in the cytoplasmic COPS5 mini-complex in hypomorphic VSMCs. Nuclear export inhibition with leptomycin attenuated the increases in PCNA and cytoplasmic p27 by COPS8 hypomorphism. Interestingly, CSN deneddylase inhibitor (COPS5i-3) did not have an effect on the proliferation of COPS8-hypo VSMCs but suppressed the PDGF-BB induced PCNA increases in CTL VSMCs. We then, constructed plasmids expressing full-length COPS5 harboring a mutation either in its nuclear export sequence (NES) or in its MPN domain. We found that genetically disabling the nuclear-export but not the deneddylase activity of COPS5 suppressed the hyperproliferation and restored p27 nuclear localization of hypomorphic VSMCs. These data suggest that promoting the nuclear export of p27 by COPS5 rather than the deneddylation activity of COPS5 is responsible for the greater PDGF-BB induced proliferation of COPS8-hypo VSMCs. Conclusion: COPS8 hypomorphism promotes the proliferation of VSMCs in injured vessels through the increased nuclear exclusion of p27 mediated by COPS5.

CSN5‐mediated nuclear exclusion of p27 in vascular smooth muscle cells contributes to the exacerbation of neointimal hyperplasia by CSN8 hypomorphism

BackgroundNeointimal hyperplasia is a common pathological response to vascular injury and is caused by the proliferation and migration of vascular smooth muscle cells (VSMCs). The COP9 signalosome (CSN) is a highly conserved protein complex composed of 8 unique protein subunits (CSN1 through CSN8) that functions as the deneddylase for Cullin and thereby regulates ubiquitination. However, CSN subunits may have deneddylation‐independent function. Downregulation of CSN8 was shown to accelerate the growth rate of cultured mouse embryonic fibroblasts but impaired cardiac protein quality control in mice. Despite the strong evidence linking CSN to cell cycle regulation, direct study of CSN in VSMC proliferation is lacking. To fill this critical gap, we performed the present study to test the hypothesis that increased CSN5‐mediated nuclear exclusion of p27 contributes to the promotion of VSMC proliferation in injured vessels by CSN8 hypomorphism (Figure 1).Methods and ResultsTo produce neointimal hyperplasia in vivo, we subjected adult CSN8 hypomorphic (CSN8‐hypo) and the littermate non‐hypomorphic control mice (CTL) to left common carotid artery (LCCA) ligation and collected the LCCA segment proximal to the ligation for analyses. LCCA ligation induced significant increases in CSN8 proteins in wild type mice (p=0.0109), suggesting an important role of the CSN in vascular pathology. Compared to CTL, the CSN8‐hypo mice displayed more severe neointimal thickening at both 1‐ and 4‐week after ligation, a markedly greater increase in PCNA (proliferating cell nuclear antigen) as revealed by western blot analyses (2.05±0.46 vs. 1.37±0.098, p=0.0214), and a significantly greater prevalence of Ki67‐positive VSMCs as detected by double immunostaining for Ki67 and Sm‐22 at 1 week after LCCA ligation (p=0.005). In cell cultures, in response to platelet derived growth factor (PDGF‐BB), CSN8‐hypo VSMCs displayed an accelerated G0/G1 progression (p=0.04) and greater proliferation as assessed by measuring PCNA (p=0.015) and MTT assays (p=0.009), compared to CTL VSMCs. Furthermore, nuclear fractionation revealed that the nuclear to cytoplasmic ratio for p27 and CSN5 proteins were markedly smaller in hypomorphic mice (p=0.0038; p= 0.0246) and hypomorphic VSMCs (p=0.0045; p=0.027); and native gel electrophoresis revealed a significant increase in the cytoplasmic CSN5 mini complex or free CSN5 by CSN8 hypomorphism (in vivo, in vitro; p&lt;0.05). Interestingly, significant increases in PCNA by CSN8 hypomorphism was attenuated by nuclear export inhibitor leptomycin B (LMB) (p=0.04) but not by CSN deneddylase inhibitor (CSN5i‐3). CSN5i‐3, however, suppressed the PDGF‐BB induced PCNA increases in control VSMCs. This suggests that nuclear export (of p27) rather than changes in CSN deneddylation activity is responsible for greater PDGF‐BB induced PCNA increases in CSN8‐hypo VSMCs.ConclusionCSN8 hypomorphism increases VSMC proliferation and promotes neointimal hyperplasia in injured arteries likely through the nuclear exclusion of p27 protein that is mediated by CSN5.

Mediastinal Lymphoma in a Cat with Feline Leukemia Virus Mediastinal Lymphoma in a Cat with Feline Leukemia Virus

Background: Mediastinal lymphoma occurs at a high incidence in cats positive for feline leukemia virus (FeLV). It is a malignant lymphocytic neoplasm that may trigger clinical signs such as dyspnea, apathy, regurgitation, and weight loss. The objective of this work is to report a case of mediastinal lymphoma associated with FeLV in a cat, and describe the clinical, pathological, diagnostic, and therapeutic aspects that can help the diagnosis and treatment of this disease. Case: A 3-year-old male neutered cat weighing 4.6 kg, positive for FeLV, and with a history of dyspnea and hyporexia was referred to a private veterinary clinic in the city of Caxias do Sul, RS, Brazil. The only alteration found at the clinical examination was a muffled sound during lung auscultation. Radiographs of the thorax revealed the presence of pleural effusion; after drainage of the fluid, a mass located in the mediastinal area became radiographically observable. An analysis of the effusion fluid showed high cellularity characterized by a markedly pleomorphic population of individual round cells consistent with lymphocytes. Small lymphocytes with a high nucleus:cytoplasm ratio, round nucleus, condensed chromatin, and inconspicuous nucleoli were observed, along with medium and large lymphocytes. The medium and large lymphocytes were characterized by a variably increased nucleus:cytoplasm ratio and a scant to moderate cytoplasm exhibiting moderate to intense basophilia and, occasionally, vacuoles. The nuclei were round, idented, or irregular; most of them were located eccentrically and contained coarse to finely granular chromatin. Nucleoli varied from single to multiple, round to angular, and central to peripheral and prominent. Macronucleoli and marked anisonucleosis were also observed, as well as binucleated cells and rare multinucleated cells. The conclusion was that it was a case of neoplastic effusion caused by a lymphoma. After an appointment with an oncologist, a chemotherapy protocol was established. The treatment of choice was CHOP, a combination of cyclophosphamide (20 mg/kg orally), doxorubicin (1 mg/kg intravenously), vincristine (0.5 mg/m2 intravenously), and prednisolone with a regressive dose starting at 2 mg/kg. The patient underwent 4 rounds of chemotherapy, and received a maintenance treatment thereafter. The patient’s condition remained stable for 390 days without relevant clinical or hematological alterations. Discussion: The diagnosis of mediastinal lymphoma was established by associating clinical finings, laboratory exams, and radiographic findings. Exams of utmost importance to reach this diagnosis were the thoracic radiography, which revealed the presence of a mass in the thorax, and the cytopathological analysis of the effusion fluid, as round cell neoplasms are easily dissociated. Young cats are the group most affected by FeLV, and mediastinal lymphoma is considered the most prevalent type of lymphoma in this species. The most common clinical sign is dyspnea; however, other signs such as apathy, weight loss, regurgitation (due to pressure on the esophagus), and Horner’s syndrome (owing to pressure on the thoracic sympathetic innervation) may also occur. This disease is treated with chemotherapy, with CHOP frequently used as the chemotherapeutic protocol. The good therapeutic response for a longer time than the average estimated by studies on the prognostic of this disease indicates that a correct diagnosis along with an assertive approach and the cooperation of the tutor are essential in cases of mediastinal lymphoma. Keywords: cat, neoplasm, virus, mediastinum, effusion. Título: Linfoma mediastinal em gato cm vírus da leucemia felina Descritores: gato, neoplasia, vírus, mediastino, efusão.

The value of ‘raspberry bodies’ in intraoperative cytologic evaluation of adnexal masses for the diagnosis of clear cell carcinoma of the ovary: A cytological-pathological correlation

We report a case of a 48-year-old female who presented to the emergency department with pelvic/abdominal pain and a recent history of irregular periods. Pelvic ultrasound and computed tomography (CT) scan of the abdomen/pelvis revealed a 7.3 cm adnexal mass with suspicious features. During the intraoperative evaluation, a frozen section slide and a cytological smear were prepared. The cytological preparation was moderately cellular, showing cohesive groups of atypical cells with anisonucleosis, high nuclear to cytoplasmic ratio, and oval nuclei with prominent nucleoli. The tumor cells surrounded extracellular, magenta hyaline globules, forming raspberry bodies. Raspberry bodies are comprised of basement membrane deposits and are a unique finding in ovarian clear cell carcinoma. Raspberry bodies were also found in the frozen section slide, but, in comparison to the cytological preparation, were rare, difficult to identify, and resembled necrotic debris. The intraoperative diagnosis of a clear cell carcinoma is important because the surgical management will be more aggressive, as optimal tumor debulking is shown to have better overall survival. In this manuscript, we detail the intraoperative evaluation of an ovarian mass, the utility of cytological preparation and importance of identifying raspberry bodies in the evaluation of ovarian masses, and surgical management of clear cell carcinoma.

Reduced volumetric parameters of the placenta and extravillous trophoblastic cells in complicated pregnancies may lead to intrauterine growth restriction and small for gestational age birth.

We undertook this study to determine quantitative changes of the placenta, focusing on extravillous trophoblastic cells (EVTs) in pregnancies with intrauterine growth restriction (IUGR) and small for gestational age (SGA) compared to the control group. Placentas from pregnancies complicated with SGA-IUGR (n=10) and control group (n=10) were obtained after cesarean surgery and evaluated using stereological assays after routine tissue processing and Masson's trichrome staining. Mann-Whitney U-test was employed, and the level of statistical significance was set at p <0.05. Our results showed that the volumetric parameters, including the total volume and volume density of chorionic villi, intervillous spaces, blood vessels in chorionic villi, and syncytiotrophoblast, decreased significantly in the SGA-IUGR group compared to control placentas (p <0.05). Also, total volume, number of EVTs, volume, the diameter of cytoplasm, and diameter of the nucleus in these cells were significantly lower in the SGA-IUGR group (p <0.05). In addition, the nucleus to cytoplasm ratio of EVTs was also higher in the SGA-IUGR group (p <0.05). There are several significant histological and stereological differences in the placenta, particularly its EVTs from the SGA-IUGR group compared to the control group. It seems that histological changes in the placental tissues could be helpful for the retrospective explanations of pregnancy complications.

Investigating the Role and Mechanism of Octreotide in Long Standing Diabetes-induced Cognitive Impairment in Rats

Abstract: Background: Octreotide is a somatostatin analogue and it produces beneficial effects in diabetic-neuropathy. Studies have also shown its beneficial role in Alzheimer disease. However, the role of octreotide in diabetes-induced cognitive impairment is not explored yet. Aim: The present study was designed to explore the role and mechanism of octreotide in long standing diabetes-induced cognitive impairment in an experimental model. Materials and Methods: Streptozotocin (60 mg/kg)-injected rats were kept for 10 weeks to induce the development of cognitive impairment, which was assessed using Morris Water Maze test. The learning was assessed by comparing the escape latency time (ELT) of day 1 with ELT of day 4. The memory was assessed on the 5th day by measuring the time spent in target quadrant (TSTQ). Three different doses of octreotide (10, 20 and 40 μg/kg) were administered for the last two weeks. The levels of Nrf2, reduced glutathione, TBARS, IL-1 and TNF-α were measured in rat brain homogenates. Results: Treatment with octreotide for two weeks led to significant increase in day 4 ELT and day 5 TSTQ in streptozotocin-injected rats suggesting the improvement in learning and memory. Moreover, octreotide attenuated streptozotocin-induced increase in neuroinflammation and oxidative stress. It also increased the nuclear: cytoplasmic ratio of Nrf2 suggesting the effect of octreotide in increasing the levels of endogenous antioxidants. Conclusion: Octreotide has the potential to improve learning and memory in long standing diabetes-induced cognitive dysfunction and its beneficial effects may be possibly attributed to decrease in neuroinflammation and oxidative stress. Key words: Oxidative stress, Diabetes, Memory, Octreotide, Neuroinflammation, Learning.

Correlation of nuclear pIGF-1R/IGF-1R and YAP/TAZ in a tissue microarray with outcomes in osteosarcoma patients.

Osteosarcoma (OS) is a genetically diverse bone cancer that lacks a consistent targetable mutation. Recent studies suggest the IGF/PI3K/mTOR pathway and YAP/TAZ paralogs regulate cell fate and proliferation in response to biomechanical cues within the tumor microenvironment. How this occurs and their implication upon osteosarcoma survival, remains poorly understood.Here, we show that IGF-1R can translocate into the nucleus, where it may act as part of a transcription factor complex. To explore the relationship between YAP/TAZ and total and nuclear phosphorylated IGF-1R (pIGF-1R), we evaluated sequential tumor sections from a 37-patient tissue microarray by confocal microscopy. Next, we examined the relationship between stained markers, clinical disease characteristics, and patient outcomes. The nuclear to cytoplasmic ratios (N:C ratio) of YAP and TAZ strongly correlated with nuclear pIGF-1R (r = 0.522, p = 0.001 for each pair). Kaplan–Meier analyses indicated that nuclear pIGF-1R predicted poor overall survival, a finding confirmed in the Cox proportional hazards model.Though additional investigation in a larger prospective study will be required to validate the prognostic accuracy of these markers, our results may have broad implications for the new class of YAP, TAZ, AXL, or TEAD inhibitors that have reached early phase clinical trials this year.

Biliary cytology: A diagnostic tree for adenocarcinoma based on a cohort of 135 patients with endoscopic retrograde cholangiopancreatography for stenosis of the extrahepatic bile duct.

The diagnosis of well-differentiated adenocarcinoma on bile aspiration is a well-known challenge. This study was aimed at improving the diagnostic performance and providing a biliary cytology learning atlas. This single-center, retrospective study included 135 cases of informative biliary samples collected between 2009 and 2018 that were classified as benign, atypical, or malignant. A double assessment was performed by a novice and a cytopathologist experienced in biliary cytology to establish the specificities, sensitivities, and inter- and intraobserver κ index agreements of 24 cytological criteria, which were illustrated in a learning atlas. A multivariate logistic regression was used to assess whether the most specific and reproducible criteria were associated with malignancy. A scoring system was statistically determined: 6 points were attributed in the presence of a 3-dimensional (3D) cluster, anisonucleosis, and a nuclear to cytoplasmic (N:C) ratio > 0.5, whereas 4 points were given in the presence of an enhanced nuclear membrane. A score higher than 10 points resulted in a malignancy diagnosis with 96% sensitivity and 97% specificity. A diagnostic tree of malignancy based on 4 criteria, together with a multidisciplinary approach, allows the diagnosis of adenocarcinoma with a specificity of 100% and a sensitivity of 88% or 72% depending on the presence of a single malignant cell or the presence of 3 combined criteria (a 3D cluster, anisonucleosis, and an N:C ratio > 0.5). It comes with a learning atlas useful for cytopathologist training and accuracy in this uncommon cytology.

Does hand mirror morphology a useful diagnostic tool for the treatment decision of acute undifferentiated leukemia at admission?

It is difficult how to manage acute undifferentiated leukemia in daily practice, but generally, hand mirror morphology provides ease to treat these patients. Thirty-nine years old male patient was admitted to with the complaints of echymosis, and pain at his left buttock due to an intramuscular injection for the treatment of previously diagnosed of the lower respiratory infection. Peripheral blood smear revealed >%50 blasts cells with a moderate nuclear: cytoplasmic ratio and one or more nucleoli. The blast cells showed a hand-mirror morphology and not harboring auer rods. According to the flow cytometric analysis the blastic cells do not represent to be originated from myeloid or lymphoid origin, because the cells harboring both of two cell lineages. AML-like therapy was commenced based on the positive myeloid markers including CD117 and CD135. Even though hand mirror morphology of the blasts usually demonstrates the lymphoid origin and the patients are treated as ALL like therapy, myeloid blasts rarely represents the same morphology, as was in our patient.

Comparative assessment of exfoliated gingival cells in healthy individuals and chronic periodontitis patients with and without tobacco chewing habit: A cytomorphometric study.

Background:Cytomorphometry is the analysis of the morphometry of exfoliated cells. Studies on exfoliated gingival epithelial cells in chronic periodontitis were not evident on the literature search. In the backdrop of this information, we were inspired to conduct a cytomorphometric study.Aim:The aim of the study is to correlate the nuclear area (NA), cytoplasm area (CA), and the nuclear: cytoplasm ratio (N/C) in the gingival cells of healthy individuals, patients with chronic periodontitis, and chronic periodontitis patients who are tobacco chewers.Materials and Methods:Ninety subjects, aged 35–45 years, were categorized into three equal groups of 30 each. Group I had individuals with healthy gingiva, Group II patients with chronic periodontitis without any deleterious habits, while Group III patients with chronic periodontitis and tobacco chewing habit. Exfoliated gingival cells were stained with Papanicolaou stain. Analysis of cells was done using binocular light microscope under ×40 and i-progress® image analyzer software.Results:There was statistically significant increase in CA in all the groups, but NA and N/C ratio did not show any statistical significant difference. When Group II was compared with Group III, we found significant correlation of CA with pocket probing depth and clinical attachment loss (CAL) in Group II. Similarly, we also found a correlation of CA and N/C with pocket depth and CA with CAL in Group III.Conclusion:The results suggest that molecular level changes occur in the cells. Cytomorphometry can be used to analyze the cellular changes in chronic periodontitis patients even before clinical signs manifest. Hence, it can be used as an investigative tool.

Development of a semi-quantitative scoring protocol for gill lesion assessment in greenlip abalone Haliotis laevigata held at elevated water temperature.

Water temperatures that exceed thermal optimal ranges (~19 to 22°C for greenlip abalone Haliotis laevigata, depending on stock genetics) can be associated with abalone mortalities. We assessed histopathological changes in H. laevigata gills held in control (22°C) or elevated (25°C) water temperature conditions for 47 d by developing a new scoring protocol that incorporates histopathological descriptions and relative score summary. Lesions were allocated to 1 of 3 reaction patterns, (1) epithelial, (2) circulatory or (3) inflammatory, and scored based on their prevalence in gill leaflets. Indices for each reaction pattern were calculated and combined to provide an overall gill index. H. laevigata held in 25°C water temperature had significantly more epithelial lifting and hemolymph channel enlargement and significantly higher gill and circulatory reaction pattern indices than H. laevigata held in 22°C water temperature. One H. laevigata had a proliferation of unidentified cells in the v-shaped skeletal rod of a gill leaflet. The unidentified cells contained enlarged nuclei, a greater nucleus:cytoplasm ratio and, in some cases, mitotic figures. This cell population could represent a region of hematopoiesis in response to hemocyte loss or migration to a lesion. Without thorough diagnostic testing, the origin of these larger cells cannot be confirmed. The new scoring protocol developed will allow the standard quantification of gill lesions for H. laevigata, specifically for heat-related conditions, and could further be adapted for other Haliotis spp.