Cell Pleomorphism Research Articles

Corneal endothelial iayer changes in various stages of keratoconus and their correlation with topographic findings: a cross-sectional study

Background. Keratoconus (KCN) is a multifactorial progressive corneal disease that leads to significant disturbances in visual function. Various layers of the cornea, such as the endothelium, are affected by this condition. This study aimed to evaluate the corneal endothelial layer in different stages of the KCN and its correlation to topographic findings. Methods. It is a cross-sectional study of 110 eyes of 70 patients (28 females and 42 males) with KCN. The KCN severity staging in this study was based on the Amsler-Krumeich criteria and topographic findings. Specular microscopy was used to evaluate and compare the condition of the endothelial layer in different stages of KCN. Results. The prevalence of the various stages of KCN was 10%, 33.6%, 13.6%, and 42.8%, respectively, for stages I to IV. There was no significant difference between various stages of OCN regarding endothelial cell density and polymegethism (P=0.759 and P=0.052, respectively). However, the mean frequency of the hexagonal endothelial cells (6A) in stage IV was 7.8% lower than in stage II eyes (P=0.025). In addition, the Kmax had a positive correlation with polymegethism (P<0.001, r2=0.110) and a negative correlation with 6A (P=0.007, r2 =0.066). Conclusion. According to our findings, the progression of KCN could be associated with increased endothelial cell pleomorphism, and higher Kmax could be associated with polymegethism. However, there was no significant difference in endothelial cell density or polymegethism between the various stages of KCN. Practical Implications. Regarding the possible changes in the corneal endothelial layer with KCN progression, the pre-operative endothelial evaluations could improve the surgical plans in these patients.

Nuclear anaplasia of positive and negative hormonal receptor (estrogen and progesterone receptor) in breast cancer cytology

Background: Breast epithelial duct cells show response to sex hormone such as estrogen and progesterone since they have receptor for estrogen and progesterone. Consequently, malignant cells from breast epithelial ductuli (Breast Ductal Carcinoma) show positivity for estrogen and progesterone receptor (ER and PR). However, when this neoplasm becomes poorly differentiated, they lost these characteristics and became ER and PR negative. On the other hand, poorly differentiated cancer cells show more nuclear enlargement that was accompanied by increase nuclear pleomorphism. This study wants to compare area of breast cancer cells nuclei and nuclear pleiomorphism in positive and negative ER and PR. Material and method: Positive and negative ER and PR breast cancer cases were collected from archive of Anatomic Pathology Department each consisting of ten to fourteen cases. Image analysis was made from each case to measure the area, diameter, and nuclear pleiomorphism. Result and discussion: There were no differences in nuclear size and pleomorphism of breast cancer cells with positive and negative ER and PR. This result show that poorly differentiated cancer cells did not accompany by increase of nuclear anaplasia. Conclusion: There is no difference of cancer cells morphology in positive and negative ER and PR. Hormonal status and anaplasia of cell nuclei can be used as independent prognostic factor to breast cancer progression.

Cell pleomorphism and changes in the enzymatic profile of selected Candida albicans strains in interaction with Escherichia coli – pilot study

Interactions between C. albicans and the microbiota play an important role in maintaining the balance between commensal and pathogenic organisms. Although the exact role of bacteria in reducing the pathogenicity of yeast remains poorly understood, a few examples have been documented so far: probiotics administration effectively reduces the formation of biofilm and bacterial metabolites inhibit the formation of hyphae.The aim of the study was to analyze C. albicans virulence levels based on the changes in the morphological structure and enzymatic profile in experimental cultures mixed with Escherichia coli. Viable cell abundance, cell pleomorphism and enzymatic profile were analyzed in single and mixed cultures (C. albicans + E. coli).The microscope analysis showed a large decrease in the number of viable C. albicans cells in mixed cultures with E. coli from 485.3±132.1 immediately after the establishment of the culture to 238.1±71.2 after an hour of incubation and 24.4±5.4 after 24 h. The length of C. albicans cells differed significantly between the single-species cultures and the mixed cultures for 24 h. Our present findings indicate a significant reduction in the secretion of several enzymes by fungi following contact with E. coli, including acid phosphatase, N-acetyl-β-glucosaminidase, naphthol-AS-BI-phosphohydrolase and leucine arylamidase.The interactions between fungi and bacteria appear to be extremely complex. On the one hand, during C. albicans with E. coli co-incubation, the bacteria stimulated the elongation of yeast cells, leading to the formation of a filamentous form; however, the number of yeast cells and their enzymatic activity decreased significantly. Therefore, it can be concluded that while E. coli stimulates some pathogenic properties, e.g. cell elongation, it also inhibits other virulence features, e.g. enzymatic activity of C. albicans.

PATIENT-DERIVED ORTHOTOPIC GRAFT MODEL OF ORAL CANCER: HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL COMPARISON.

Objective Patient-derived xenograft (PDX) model seems to recapitulate the histopathology of the original donor tumor. Here we compared the histopathological characteristics of the donor tumor with sequential implantations of tumor fragments derived from a human tongue squamous cell carcinoma biopsy in the PDX model. Study Design We orthotopically transplanted a primary tongue tumor into immunosuppressed mice, which was successively reimplanted over 6 passages (P0, P1, P2, P3, P4, and P5). Histopathological characteristics were evaluated by hematoxylin and eosin staining and pan-cytokeratin; Ki-67, and HIF-1α were evaluated by immunohistochemistry. Results Tumor size was smaller in later passages (P4 and P5). P0 tumors were classified as well-differentiated, while the other passages were classified as moderately differentiated. Vascular and perineural invasion occurred only in later passages and were not present in the donor patient tissue. Dyskeratosis and keratin pearls were observed in most of the studied passages but not evident in the patient sample. Cell pleomorphism was similar between the studied passages. Ki-67 showed very homogeneous cell proliferation rates between passages and the donor tumor. HIF-1α immunolabeling was absent in the analyzed tumor samples. Conclusion P1 and P2 tumors more accurately reproduce the morphological characteristics of the donor patient's primary tumor. (FAPESP 2019/21513-2) Patient-derived xenograft (PDX) model seems to recapitulate the histopathology of the original donor tumor. Here we compared the histopathological characteristics of the donor tumor with sequential implantations of tumor fragments derived from a human tongue squamous cell carcinoma biopsy in the PDX model. We orthotopically transplanted a primary tongue tumor into immunosuppressed mice, which was successively reimplanted over 6 passages (P0, P1, P2, P3, P4, and P5). Histopathological characteristics were evaluated by hematoxylin and eosin staining and pan-cytokeratin; Ki-67, and HIF-1α were evaluated by immunohistochemistry. Tumor size was smaller in later passages (P4 and P5). P0 tumors were classified as well-differentiated, while the other passages were classified as moderately differentiated. Vascular and perineural invasion occurred only in later passages and were not present in the donor patient tissue. Dyskeratosis and keratin pearls were observed in most of the studied passages but not evident in the patient sample. Cell pleomorphism was similar between the studied passages. Ki-67 showed very homogeneous cell proliferation rates between passages and the donor tumor. HIF-1α immunolabeling was absent in the analyzed tumor samples. P1 and P2 tumors more accurately reproduce the morphological characteristics of the donor patient's primary tumor.

Proteomic analysis identifies HMGA2 as a novel biomarker of overall survival in papillary renal cell carcinoma.

Although papillary renal cell carcinoma (PRCC) has a relatively favorable prognosis, a small number of patients with lymph node or distant metastasis have a poor prognosis. Owing to the complex typing and heterogeneity of PRCC, it remains difficult to provide risk stratification. The objective of our research was to identify potential markers of PRCC prognosis. We performed proteomics and bioinformatics analyses on six pairs of formalin-fixed paraffin-embedded tumor and paired normal tissue samples. The Cancer Genome Atlas (TCGA) data were used to analyze the prognostic value of differentially expressed proteins (DEPs) in PRCC. We verified the expression of the major biomarker through immunohistochemistry (IHC) in 91 PRCC tumor specimens. Proteomic analysis revealed 1544 DEPs between tumor and paired normal tissues. PRCC transcriptomic data from the TCGA database revealed that compared to non-tumor tissues, the expression of high-mobility group protein A2 (HMGA2) was upregulated in tumor tissues, and patients with high HMGA2 expression exhibited shorter overall survival times. HMGA2 was associated with PRCC tissue subtype and higher cell pleomorphism. Both TCGA and IHC results showed that HMGA2 expression was associated with lymph node metastasis and clinical stage. HMGA2 was positively correlated with malignant progression and could be a valuable novel prognostic biomarker for PRCC risk stratification.

Different Course of Immune Reactions and Endothelial Cell Loss after Penetrating Low-Risk Keratoplasty and Descemet Membrane Endothelial Keratoplasty for Fuchs Endothelial Dystrophy.

The aim of this study was to compare the incidence of immune reactions and endothelial cell loss after penetrating keratoplasty (PKP) vs. Descemet membrane endothelial keratoplasty (DMEK) in patients with Fuchs endothelial dystrophy (FED). In the present retrospective study, a total of 962 surgeries (225 excimer laser PKP and 727 DMEK) of 700 patients performed between 28.06.2007 and 27.08.2020 in the Department of Ophthalmology at Saarland University Medical Center UKS were statistically evaluated. On the one hand, the prevalence and the temporal course of the immune reactions that occurred were analysed using the Kaplan-Meier method, as well as the effect of the immune reactions on the endothelial cells and corneal thickness. Secondly, endothelial cell density, pleomorphism, and polymegethism of the endothelial cells were evaluated for the time points U1 = preoperative, U2 = 6 weeks postoperative, U3 = 6 to 9months postoperative, U4 = 1 to 2 years postoperative, and U5 = 5 years postoperative. In addition, statistical tests were carried out for differences between the two types of surgery and in the longitudinal course. A total of 54 immune reactions occurred during the observed period, whereby the probability of such a reaction was significantly greater in the PKP group with 8.9% than in the DMEK group with 4.5% (p = 0.011). The comparison of the two Kaplan-Meier curves also showed a significant difference between the two surgical techniques in the log-rank test (p = 0.012). The endothelial cell loss due to the immune reaction was only significant in PKP (p = 0.003). For all surgical procedures, endothelial cell density decreased significantly with time in both surgical techniques (p < 0.0001 in each case), but more strongly with DMEK than with PKP (p < 0.0001). Furthermore, this cell density was significantly higher with PKP than with DMEK for the whole observation time (p < 0.0001). Polymegethism decreased significantly in the DMEK group (p < 0.0001). Pleomorphism was significantly higher, on average, in DMEK than in PKP (p < 0.0001). The prognosis of DMEK in patients with FED seems to be more favourable after immune reactions than that of PKP, as not only were immune reactions less frequent, but they were also milder. However, endothelial cell density was significantly higher in the PKP group during the entire follow-up.

Solitary, subcutaneous, fixed, firm, and fast-growing nodule

Solitary, subcutaneous, fixed, firm, and fast-growing nodule

Morphological Changes in Major Salivary Glands in Mice Treated With a Choline and Methionine Deficient Diet.

It has been shown that the methionine-choline deficient (MCD) diet induces hepatocarcinogenesis, but not in extrahepatic organs, such as the testis, and pancreas, although may increase chemical-induced carcinogenesis in the colon, mammary gland, esophagus, and pancreas. Accumulating evidence suggests that salivary glands are very susceptible to stress conditions, such as radiation, hyperglycemia, and exposure to xenobiotics in vivo. This study aimed to analyze the histological changes on the major salivary glands (parotid, submandibular, and sublingual) after MCD diet administration. Male Swiss mice were submitted to ad libitum access to the control (AIN-76) or MCD diet for 28 days. The rebound group received the MCD diet for 24 days and the control diet for 10 days. Using the AxioImager A2 microscope, the hematoxylin-eosin (HE) stained specimens (4 mm) were evaluated for tissue degeneration, nuclear hyperchromatism and atrophy. In the parotid gland from the MCD group, tissue degeneration, pyknosis, apoptosis and atrophy were observed, which remained in the rebound group, associated with hyperchromatism. In the submandibular gland from both MCD and rebound groups, severe tissue disorganization was associated with cell pleomorphism, hyperchromatic cells, apoptosis, increased eosinophilia, and inflammatory infiltrate. Finally, in the sublingual gland, there were no histological alterations in the experimental groups compared to the control. MCD can induce pre-neoplastic changes in the mouse parotid and submandibular glands, which are not reversed by a change in the diet.

Segmentation of endothelial cells of the cornea from the distance map of confocal microscope images

We propose a novel algorithm for segmenting cells of the cornea endothelium layer on confocal microscope images. To get an inter-cellular space with minimum gray-scale value and to enhance cell borders, we apply a difference of Gaussian filter before image binarization by thresholding with the minimum gray-scale value. Removal of segmented noise and artifacts is performed by automatic thresholding (using an image frequency analysis to obtain a global threshold value per image). Final segmentation of cells is achieved by fitting the largest inscribed circles into the centers of cell regions defined by the distance map of the binary images. Parameters of interest such as cell count and density, pleomorphism, polymegathism, and F-measure are computed on a publicly available data-set (Confocal Corneal Endothelial Microscopy Data Set - Rotterdam Ophthalmic Data Repository) and compared against the results of the segmentation methods included with the data set, and the results of state of the art automatic methods. The obtained results achieve higher accuracy compared to the results of the segmentation included with the data set (e.g., -proposed versus dataset in R2 and mean relative error-, cell count: 0.823, − 0.241 versus 0.017, 0.534; cell density: 0.933, − 0.067 versus 0.154, 0.639; cell polymegathism: 0.652, − 0.079 versus 0.075, 0.886; cell pleomorphism: 0.242, − 0.128 versus 0.0352, − 0.222, respectively), and are in good agreement with the results of the state of the art method.

Pathological and immunohistochemical study of colon cancer. Evaluation of markers for colon cancer stem cells.

Colorectal cancer is a major public health problem worldwide with increasing morbidity and mortality. Numerous exogenous and endogenous factors are involved in colorectal carcinogenesis: age, sex, diet, smoking, alcohol consumption, exposure to harmful environmental factors, intestinal microbiota, bacterial and viral infections, the ability of the host immune system to respond, genetic factors, etc. The present study analyzed histopathologically and immunohistochemically a number of 36 cases of colorectal adenocarcinomas. The existence of an accentuated cell pleomorphism was noted, which corresponds to different clones of tumor cells, in the same tumor coexisting aspects of tubular adenocarcinoma, mucinous areas and even signet-ring cell. The tumor stroma was mainly of the desmoplastic type, but also of the lax type, more or less infiltrated with inflammatory cells. Evaluation of immunomarkers for cancer stem cells (CSCs) showed that none of the markers used alone [cluster of differentiation (CD)133, CD44, aldehyde dehydrogenase 1 family member A1 (ALDH1A1), CD24, CD26] show CSCs.

Polyostotic osteosarcoma associated with avian leukosis virus infection in a captive bare\u2010faced curassow (Crax fasciolata)

BackgroundOsteosarcoma is a malignant mesenchymal bone tumor. Although it is a common tumor in the appendicular skeleton of dogs and cats, it is rarely reported in birds. Retroviruses are usually associated with solid tumor development in different avian species.Case presentation:This report aims to describe a case of osteosarcoma associated with the avian leukosis virus in a captive bare-faced curassow (Crax fasciolata). A captive adult female bare-faced curassow presented with lameness, hyporexia, and a non-ulcerative and firm tumor in the right femur. The bird was euthanized due to the poor prognosis. Histopathology revealed an infiltrative mesenchymal neoplasm consisting of spindle cells with moderate cell pleomorphism, organized in bundles and interspersed by marked deposition of the osteoid matrix, which was compatible with osteosarcoma affecting both femur and tibiotarsus, with renal metastasis. Immunohistochemistry of the primary and metastatic tumor demonstrated vimentin expression by neoplastic cells. Samples of the neoplasm, bone marrow, and spleen were processed for PCR, which enabled the demonstration of proviral avian leukosis virus (ALV) DNA.ConclusionsTo the best of our knowledge, this is the first report of an osteosarcoma in a bare-faced curassow with an unusual polyostotic manifestation and associated with ALV infection.

Pleomorphic Fibroma: A Clinicopathologic Case Series With the Review of the Literature.

Pleomorphic fibromas are rare flesh-colored benign neoplasms first described in 1989. Their histopathology is notable for nuclear pleomorphism of spindle cells and multinucleate giant cells but lacking mitoses. The cellular origin of these tumors is unknown. This case series describes an additional 18 lesions with discussion of histopathology and immunohistochemistry. This case series of 18 pleomorphic fibromas uses immunohistochemical staining for CD34, CD68, factor XIIIa, and S-100 and general histopathologic examination under light microscopy to describe the lesions. Immunohistochemical stains for CD34 showed nearly universal positivity of the pleomorphic spindle cells, although some more focally. The pleomorphic cells were negative for CD68, variably positive for factor XIIIa, and universally negative for S-100. All the lesions showed characteristic nuclear pleomorphism with absent mitoses. Collagen thickening was variable, mucin was absent, and perivascular inflammation was present in all lesions. Pleomorphic fibromas are fibrous lesions with benign clinical course and histopathologic findings including nuclear pleomorphism. Immunohistochemical staining characteristics of the lesion, along with unique spindle cells and multinucleate giant cells help to differentiate this from other tumors.

Squamous Cell Carcinoma in the Third Eyelid of a Dog

Background: Squamous cell carcinoma is a malignant neoplasm that originates from the keratinized stratified squamous epithelium and predominantly affect light-skinned animals. In dogs, breeds such as American Staffordshire Terriers, white or speckled Bull Terriers, and Beagles have a higher predisposition. Squamous cell carcinoma presents in the skin, at slightly pigmented or hairy sites, especially in digits, but also may occur in the nasal planum, oral mucosa, and rarely, in the eye. Considering that few reports have been published on eye neoplasms, the aim of this paper is to describe a dog with a lesion in the third eyelid of his right eye which was diagnosticated with squamous cell carcinoma. Case: A 10-year-old male American Staffordshire dog was admitted to the Veterinary Medical Teaching Hospital of the Veterinary Medicine and Zootechnics College, Universidade Federal de Mato Grosso do Sul (UFMS), Campo Grande, Brazil with injury to the right eye. During the physical examination, there was also a non-adhered lump near the foreskin, measuring 1.5 cm in diameter. In addition, there was another lump in the third eyelid of the right eye, approximately 3 mm in diameter. Cytology of the dermal nodule was performed by fine-needle aspiration cytology; however, the sample was insufficient for cytological evaluation. Therefore, the animal was placed under general anesthesia for skin lump excision and for fine-needle aspiration cytology of the third eyelid nodule. The histopathological exam revealed high cellularity of epithelial cells, intense anisocytosis and pleomorphism, cytoplasmic basophilia and vacuolation, multiple evident nucleoli, and anisocariosis and coarse chromatin. These finds were compatible with squamous cell carcinoma, which was the same result suggested by fine-needle aspiration cytology of the third eyelid sample. Based on these results, the dog underwent a surgical procedure for enucleation and subsequent histopathological evaluation of the nodule in the third eyelid, which confirmed the squamous cell carcinoma diagnosis.Discussion: Squamous cell carcinoma is an extremely aggressive tumor with low metastatic potential, characterized by invasion of the dermis by proliferation of malignant epithelial cells from the prickly layer. It is most common in elderly animals, and American Staffordshires are among the breeds that are predisposed to develop this tumor. The clinical presentation is highly variable, depending on the tissue involved. In this case, the dermal nodule was an elevated area on the skin and the third eyelid nodule resembled an ulcerative mass. Cytological examination from the lesion located on the third eyelid, showed malignancies cytoplasmic changes frequently found in carcinomas such as anisocytosis, cytoplasmic basophilia, and cell pleomorphism. In addition, nuclear changes had also occurred, such as crass chromatin, multiple evident nucleoli, and multinucleated cells. A presumptive diagnosis was made based on cytology and was confirmed after biopsy and histopathological examination. Because it is uncommon in dogs, squamous cell carcinoma of the third eyelid may be misdiagnosed, delaying correct treatment, and accelerating the development of the tumor. Currently, various therapeutic approaches are available, such as surgical excision, electrosurgery, cryosurgery, radiation, and hyperthermia. The choice of treatment depends on the location and stage of the lesions. Surgical treatment should be aimed at removing sufficient tissue to leave surgical margins free of neoplastic cells.

Cardiac Tamponade Secondary to Hemangiosarcoma in a Dog

Background: Nonspecific clinical manifestations such as apathy, anorexia and diarrhea are common in the clinical routine, and therefore may mask the severity of its triggering factor. When patients presenting this symptomatology are referred to the care center, it is essential that a thorough investigation is performed to clarify the primary causes of these manifestations, and for this, complementary imaging exams may be necessary. The objective of this study is to describe the clinical and imaging aspects of a canine with cardiac hemangiosarcoma and to correlate with the pathophysiology of the alterations observed, in order to optimize the clinical care of patients with nonspecific clinical signs and affected by this alteration.Case: A 10-year-old Pitbull dog was treated with a history of vomiting, loss of appetite and diarrhea. Upon clinical examination, the animal presented dehydration level and distended abdomen. On ultrasound examination, hepatomegaly was observed, associated with signs of double layer in the gallbladder wall and the presence of moderate peritoneal effusion. On chest radiography, the cardiac silhouette showed an important increase in size with loss of shape and contours. Dorsal displacement of the trachea, greater contact of the heart with the sternum and displacement of the caudal vena cava were observed. In addition, an amorphous and poorly defined structure with radiopacity of soft tissues was observed in the region of the atrium and right ventricle, causing the obliteration of pulmonary fields. On echocardiographic examination, a significant amount of free anechogenic fluid was observed in the pericardial sac, confirming the suspicion of pericardial effusion, with consequent collapse of the wall of the right heart chambers during systole. Furthermore, an amorphous, poorly defined and heterogeneous structure was observed in the right atrium wall. Due to the location of the neoformation, the patient's general clinical condition, euthanasia was chosen. During the necropsy, the ocular, oral and preputial mucosae were moderately pale. In the abdomen, the presence of hydroperitoneum and hepatomegaly was observed. In the thoracic cavity, hemothorax, enlarged heart were identified and, in the right atrium, a reddish mass was identified. In addition, the lungs were whitish and hypercrepitating at the edges of the cranial lobes; the rest of the organ was moderately hyperemic, hypocrepitating. In histopathological examination of the liver, the centrilobular region showed chronic passive congestion associated with necrosis and multifocal degeneration of hepatocytes. Neoplastic proliferation of mesenchymal cells, moderately cellular, non-encapsulated, of infiltrative growth was observed in the heart. Neoplastic cells formed bundles, with a tendency to organize themselves into small blood vessels filled with red blood cells. The cytoplasm was moderate, elongated, indistinct, eosinophilic and homogeneous. The nucleus was large, unique, elongated, with finely dotted chromatin and sometimes with one or two nucleoli evident. Anisocytosis, anisocariosis and cell pleomorphism were moderate. Interwoven with neoplastic cells, a moderate presence of multifocal lymphohistioplasmocytic inflammatory infiltrate was observed. The definitive diagnosis of cardiac changes was hemangiosarcoma.Discussion: Hemangiosarcoma is a vascular endothelial cell neoplasm with high metastatic power and unfavorable prognosis. When located in the heart, it is commonly found in the auricle and right atrium and the cardiovascular changes caused by this neoplasm as well as the severity of these changes vary according to size and location. Clinical manifestations can be quite nonspecific and are usually associated with hemodynamic impairment, causing signs of right or left congestive heart failure.Keywords: heart, neoplasm, pathophysiology.Descritores: coração, neoplasia, patofisiologia.Título: Tamponamento cardíaco secundário ao hemangiossarcoma em cães

Repressed Wnt Signaling Accelerates the Aging Process in Mouse Eyes.

Purpose Ocular aging is a natural process of functional decline in vision. When the process reaches a point that compromised vision affects normal daily activity, it manifests as age-related ocular diseases, such as age-related macular degeneration, cataracts, glaucoma, and pseudoexfoliation syndrome. We previously reported that repressed Wnt signaling accelerated the maturation of corneal epithelium during tissue development. Here, we explore the hypothesis that repressed Wnt signaling is associated with accelerated aging in mouse eyes. Methods Wnt ligand antagonist secreted frizzled-related protein 1 (sFRP1) was expressed in the corneal stroma by a tissue-specific, inducible, bitransgenic system. Tissue structure was analyzed for signs of aging. Signal transduction analysis was performed to determine the cellular response to sFRP1. Results Mouse eyes with sFRP1 expression showed signs of accelerated aging, resembling those found in pseudoexfoliation (PEX) syndrome, a known age-related disease. Specific findings include granular deposition on the surface of the anterior lens capsule, pigment loss from the anterior surface of the iris, the presence of fibrillary material in the anterior chamber, and changes in cell size (polymegethism) and shape (pleomorphism) of the corneal endothelial cells. In vitro studies demonstrated that sFRP1 did not inhibit Wnt5a function and that cells responded to sFRP1 and Wnt5a in a very similar manner. Conclusion The expression of sFRP1 accelerates the aging process in mouse eyes and future studies are warranted to elucidate the underlying mechanisms.

Effect of intracameral human cord blood-derived stem cells on lasered rabbit trabecular meshwork.

This study aimed to investigate the effect of intracameral human cord blood stem cells on lasered rabbit trabecular meshwork. Immediately following diode laser application to the trabecular meshwork, human cord blood stem cells were injected intracamerally, in one eye of 12 albino rabbits. The other eye of ten rabbits was lasered controls and two eyes were normal controls. Rabbits were killed after 4, 8 and 12weeks. Lasered control rabbit eyes showed significant disruption of trabecular architecture, loss and pleomorphism of trabecular endothelial cells and progressive narrowing of trabecular spaces till 12weeks. In contrast, lasered eyes, concurrently injected with human cord blood stem cells, showed relatively preserved endothelial cellularity and structure of the trabecular meshwork, at all time points. Human CD34- and CD44-positive cells were identified in 7/8 eyes treated with stem cells, at 4 and 8weeks, and 2 of 3 at 12weeks. Many PKH26-labeled human cord blood cells were visible throughout the trabecular area at 4weeks. They gradually decreased in number by 8weeks, and at 12weeks, they appeared to be oriented along trabecular beams. There was a relative preservation of cellularity and architecture of the trabecular meshwork in eyes injected with human cord blood stem cells, as compared to lasered control eyes up to 12weeks, without significant inflammation. This suggests a probable role for such stem cells in eyes with glaucoma, having trabecular dysfunction.

Histopathological findings in the early diagnosis of granulosa cell tumour in bitches.

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call-Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call-Exner bodies and the degree of tumour cell pleomorphism (p=0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p=0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p=0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call-Exner body.

Abstract GS5-05: Resistance to neoadjuvant chemotherapy in triple negative breast cancer mediated by a reversible drug-tolerant state

Abstract Approximately 50% of patients with localized triple negative breast cancer (TNBC) have substantial residual cancer burden following treatment with neoadjuvant chemotherapy (NACT), resulting in distant metastasis and death for most of these patients. While genomic and phenotypic intra-tumor heterogeneity are pervasive features of TNBCs at the time of diagnosis, the functional contributions of heterogeneous tumor cell populations to chemoresistance have not been elucidated. To investigate tumor evolution accompanying NACT, we employed orthotopic patient-derived xenograft (PDX) models of treatment-naïve TNBC, which retain intra-tumor heterogeneity characteristic of human TNBC. We discovered that some PDX models initially exhibited partial sensitivity to standard front-line NACT (Adriamycin plus Cytoxan, AC). Following AC, residual tumors were resistant to chemotherapy but repopulated tumors with chemo-sensitive cells if left untreated, indicating that tumor cells possessed inherent plasticity. To identify the tumor cell subpopulation(s) conferring chemoresistance, we conducted barcode-mediated clonal tracking in three independent PDX models by introducing a high-complexity pooled lentiviral barcode library into PDX tumor cells which were then orthotopically engrafted into recipient mice. Strikingly, residual tumors maintained the same heterogeneous clonal architecture as naïve tumors. Concordantly, whole-exome sequencing revealed conservation of genomic subclonal architecture throughout treatment. These results were corroborated by genomic sequencing of serial biopsies pre- and post-AC obtained directly from TNBC patients enrolled on an ongoing clinical trial at MD Anderson (ARTEMIS; NCT02276443). Together, these studies revealed that genomically distinct pre-treatment subclones were equally capable of surviving AC to reconstitute tumors after treatment. To identify functional addictions of residual tumor cells, we conducted histologic and transcriptomic profiling. Residual tumors following AC-treatment exhibited extensive fibrotic desmoplasia and tumor cell pleomorphism in both PDX models and in serial biopsies obtained from TNBC patients enrolled on the ARTEMIS trial. Strikingly, these AC-induced features were reverted upon regrowth of residual tumors in PDXs and in patients' tumors. Similarly, residual tumors exhibited unique transcriptomic features, many of which are also de-regulated in cohorts of human TNBCs undergoing chemotherapy treatment. These features were nearly completely reverted after tumors regrew, suggesting that the residual tumor state may be a unique and transient therapeutic window. Gene set enrichment analyses revealed that residual tumors had increased activation of oxidative phosphorylation and decreased glycolytic signaling. Pharmacologic targeting of oxidative phosphorylation with a small-molecule inhibitor of mitochondrial electron transport chain complex I (IACS-010759) significantly delayed the regrowth of AC-treated residual tumors in three independent PDX models. Collectively, these studies reveal that a reversible phenotypic state can confer chemoresistance in the absence of genomic selection and that the residual tumor state is a novel therapeutic window for chemo-refractory TNBC. Citation Format: Echeverria GV, Ge Z, Seth S, Jeter-Jones SL, Zhang X, Zhou X, Cai S, Tu Y, McCoy A, Peoples M, Lau R, Shao J, Sun Y, Bristow C, Carugo A, Ma X, Harris A, Wu Y, Moulder S, Symmans WF, Marszalek JR, Heffernan TP, Chang JT, Piwnica-Worms H. Resistance to neoadjuvant chemotherapy in triple negative breast cancer mediated by a reversible drug-tolerant state [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr GS5-05.

Comparison of anterior segment parameters and corneal endothelial changes in patients with pseudoexfoliative glaucoma and pseudoexfoliation syndrome

Aim: The aim of this study is to evaluate the changes in anterior chamber morphology and corneal endothelium in patients with pseudoexfoliation syndrome (PEX) and pseudoexfoliative glaucoma (PEXG) and to determine the differences according to the control group. Material and Methods: 61 eyes of 61 PEX patients, 56 eyes of 56 PEXG patients and 46 eyes of 46 healthy subjects were included in the study. Measurement of intraocular pressure (IOP) was taken and axial length (AL), K1, K2, central corneal thickness (CCT), white-to-white distance (WTW), anterior chamber depth (ACD), pupil diameter (PD) by optical biometry and endothelial cell density (ECD), polimegatism (CV), pleomorphism (HEX) and average cell area (AVG) by specular microscope were taken of all the patients. Evaluation among the groups were made by ANOVA test, and p 0.05). There were no statistically significant differences among the groups in terms of IOP, AL, K1, K2, WTW, ACD (p> 0.05). Pupil diameter values were lower in the PEX group. CCT and endothelial cell counts were found to be higher in the control group compared to the other groups, while no significant difference was found between PEX and PEXG groups. AVG results were found to be significantly higher in the PEXG group than in the control group (p

Tumor cell seeding in the biopsy tract and its clinical significance in osteosarcomas.

The reasons behind the removal of the biopsy tract in osteosarcomas are not well known, and studies about tumor seeding in the biopsy tract have generated conflicting results. This study is designed to evaluate the presence and possible clinical significance of tumor cell seeding in the biopsy tract of osteosarcomas. We prospectively evaluated 55 cases of osteosarcomas for tumor cell seeding and other clinical and pathologic prognostic parameters. Eleven cases (20%) involved microscopic tumor foci in the biopsy tract. The higher local recurrence rates (P = 0.005) and worse recurrence-free survivals ( P = 0.009) were observed in patients with tumor cell foci in the biopsy tract. Mitotic rate, tumor cell pleomorphism, and matrix production in main tumor foci were higher in cases with tumor seeding ( P = 0.047, P = 0.012, and P = 0.005, respectively) CONCLUSIONS: Tumor seeding in the biopsy tract is a fact in osteosarcomas. The higher local recurrence rates are more likely to occur in cases with tumor seeding.