Journal of Cytology & Histology

Background: Damage to the myocardium following a myocardial infarction can be severe and possibly non
reversible leading to heart failure. However, recent evidence suggests that cardiomyocyte progenitor cells may be able to assist in healing the injured cells. The study purpose is to investigate the use of “Stage Specific Embryonic Antigen 1” (SSEA-1) surface marker to isolate cardiac progenitors from the human embryonic heart.

Materials and Methods: The surface marker SSEA-1 was used to isolate cardiac progenitor cells from human embryonic hearts obtained from abortion material. The fast and reliable Fluorescence Activated Cell Sorting (FACS) technique was used to sort out pure population of SSEA-1+ cells. Cells were then cultured and characterized for multipotent stem cells markers using qRT-PCR and flow cytometry. The cardiopoietic capacity of SSEA-1+ cells was evaluated using a 5-azacytidine differentiation protocol.

Results: Isolated SSEA-1+ cells expressed the pluripotent stem cell transcription factor (Oct4) as well as the
multipotent cardiac progenitor cell transcription factors (Nkx2.5, Isl-1 and Tbx5) at the mRNA level, but they did not express the mature cardiomyocyte marker-troponin T (TnT). Furthermore, these cells were found to co-express the mesenchymal stem cells markers (CD105, CD166, CD73, CD59 and CD44), but not the hematopoietic markers (CD45, CD133 and CD34). The cultured SSEA-1+ cells were cardiopoietic and responded to 5-azacytidine treatment by differentiating into cardiomyocytes with high expression of TnT and actin as well demonstrating contractile filaments on transmission electron microscopy.

Conclusions: These data demonstrate that isolated SSEA-1+ from human fetal hearts using the FACS method might be used as a template for the generation of cardiac progenitor cells from other stem cells sources. This knowledge assists in the expansion of our knowledge on the regeneration of damaged myocardium.

The fish seed resource is a major input for fish farming. There is a playing important role for increasing fisheries
production, providing food and income as well as contributing to national economy. Fishes is a delicious and
most important source of animal protein food. Since, the fish seed resource is an essential for food security, rural development and poverty alleviation. In the present investigation fin fish eggs belonging to 43 species of finfishes belong to 31 genera, 19 families, 8 orders, have been collected, identified and described in detail. Among the various finfish eggs collected, higher number of eggs were recorded for Cluepeids fishes, followed by Engraulids, Carangids, Cynoglossids, Tetraodontids, Teraponis, Mugilids, Atherinids, Synodontids, Ophichthids, Pristigastrids, Chirocentrids, Scombrids, Gerreids, Soleids, Bothids, Pleuronectids and Hemiramphids from Muthupettai waters.

Six nuclear parameters (number of focus layers, pixel count, three-dimensional luminance coefficient variation (3D-CV), roundness, intranuclear inclusion, and nuclear groove) were measured to examine the usefulness of discriminant analysis between papillary carcinoma of the thyroid (Papillary ca) or adenomatous nodule/goiter (A. nodule) using Mahalanobis distance.
 

Forty-two of 442 indeterminate specimens were histologically diagnosed with Papillary ca or A. nodule. Six
nuclear parameters for each specimen were examined to conduct discriminant analysis between papillary ca and A. nodule using Mahalanobis distance.
 

The number of nuclear focus layers, pixel count, and intranuclear inclusion [×10-1] of Papillary ca were higher than those of A. nodule (3.91 ± 0.44, 888 ± 257, and 2.2 ± 4.2 vs. 3.59 ± 0.54, 785 ± 111, and 1.3 ± 3.5). The 3DCV [×10-3], roundness [×10-1], and nuclear groove [×10-1] of Papillary ca were lower than those of A. nodule (12.1 ± 5.7, 8.464 ± 0.144, and 2.2 ± 4.2 vs. 15.1 ± 5.2, 8.602 ± 0.078, and 3.3 ± 4.9). Positive discrimination rates of each specimen were 96.3% and 73.3% for Papillary ca and A. nodule, respectively.
 

The discriminant analysis for each specimen is reasonable and useful for providing high discrimination rates.

Different forms of Aluminum are environmental xenobiotics that induce free radical-mediated cytotoxicity and
reproductive toxicity. We studied the aluminum trichloride (AlCl3) toxicity on adult rat testes. Male Wistar rats were treated orally with AlCl3 at three doses: 100, 200 and 400 mg/kg bw for 6, 12 and 18 months. Body and testes weight were followed. Plasma and testicular metal levels were assayed by atomic absorption spectrophotometry with graphic oven method. Light microscopy observation of testes was also performed. The results showed a significant decrease (p<0.05) of body weight only at 18 months with the highest dose compared to the control group. We also noted a decrease in testicular weight in all treated groups at 12 and 18 months and only for the group treated
with 400 mg/kg bw at 6 months. Aluminum trichloride resulted in a significant increase (p<0.05) plasma aluminum concentrations is proportional at doses administered at different periods. In contrast, a significant increase (p<0.05) of the testicular aluminum concentration was observed at 6 months in the group treated with 400 mg/kg bw, at 12 months in all groups and at 18 months only in groups treated with 100 and 200 mg/kg bw. Compared to the control group, a morphological alteration of the seminiferous tubules was observed at 6 months in group treated with 200 and 400 mg/kg bw, at 12 months with 200 and 400 mg/kg bw and at 18 months in all groups. Disruption of spermatogenesis was observed at 6 months with 200 and 400 mg/kg bw and at 12 and 18 months with all doses.

In summary, this study highlights the toxic effect of aluminum on the testes of adult rats. This toxicity is dose and period dependent.

We experienced 5 cases of clear cell adenocarcinoma (CCAC) of the cervix in the 10-year period from 2003 to
2012. Because a specimen from one of the cases was poorly preserved, we analyzed cytological features of the
4 remaining cases. Characteristic large, pale-staining atypical cells with prominent, round nuclei were seen in all
cases. Additionally, a hobnail appearance and basement membrane-like substance were detected in 3 cases, and a mirror ball cluster-like morphology was found in 1 case. Cervical CCAC has been reported to be difficult to diagnose by cytology; however, we were able to diagnose these 4 cases by recognizing several characteristic cytological features.

Soluble proteins of ungerminated conidia of Erysiphe necator exhibited esterase and cutinase activities, and such hydrolytic activities were measured in the parietal protein fraction. However, histochemical localisation of esterase activity was detected upon further fungal development, as well in elongating germ tubes and in appressoria. The esterase activity was spectrophotometrically quantified using para-nitrophenylbutyrate as a substrate, and cutinase activity was determined using 3H-labelled cutin. Histochemical localisation was determined using indoxyl acetate as a substrate. Diisopropyl fluorophosphate was used as inhibitor of serine hydrolase. The role of a putative constitutive parietal cutinase in the ungerminated conidia of E. necator in adhesion to the host and differentiation of infective structures, as well as implications for successful penetration, are discussed.

The Wnt-beta-catenin pathway (WP) regulates different aspects of cell fate, migration and polarity. Inappropriate deregulation of this pathway leads to oncogenic and metastatic changes. The Triple Negative (TN) subset of BC (the absence of hormone receptors and absence of amplification/overexpression of HER2 receptors) exhibits aggressive clinical behavior and has poor clinical outcome. We have recently identified WP as one of the key signature pathways in TNBC associated with metastasis [1,2]. Here we report for the first time that WP controls metastasis-associated phenotype of Vascular Mimicry (VM) in TNBC. We have established 2D and 3D models to study VM in this subset of BC and characterized different TNBC cell lines in terms of VM and identified the role of WP in the regulation of VM. To establish the role of WP in the regulation of VM in TNBC, we have used (1) a pharmacological inhibitor of beta-catenin which is a functional readout of WP and (2) signaling modulators of WP. Since PI3K-pathway is an upstream regulator of WP, we have also used inhibitors of PI3K-pathway to test our hypothesis. Mechanistically (1) sulindac sulfide (pharmacological inhibitor of beta-catenin) and XAV939 (modulator of WP) mediated decrease in beta-catenin, (2) XAV939 mediated increase in axin and (3) LY294002 or SF1126 (pan PI3K inhibitors) mediated decrease in pGSK3beta caused an abrogation of VM in various TNBC cell lines including BT20, HCC1937, SUM149,
MDA-MB231 and MDA-MB468. Finally we obtained genetic proof-of-concept using beta-catenin SiRNA. SiRNAmediated downregulation of WP abrogated VM demonstrating the involvement of WP in VM. Functionally, our data show that VM in brain specific metastatic TNBC cells is mediated via activation of WP.

Introduction: Management of nipple discharge (ND) is variable reflecting the lack of a uniformly accepted
assessment policy. The aim of this study was to determine the common underlying pathology in patients presenting with ND as well as to evaluate the diagnostic role of red blood cells (RBC) and epithelial cells in the nipple cytology (NC).

Methodology: Electronic data base held at Leeds Teaching Hospitals NHS Trust was searched to identify all
the patients who underwent Hadfield’s (open diagnostic major duct excision) procedure for ND from January 2004 to December 2009. We included only patients with ND as the primary symptom and excluded those with any underlying palpable or radiological abnormality.

Results: Hadfield’s procedure was performed in 120 cases (Median age=49 years; range=20-88). The final
histology showed benign changes (duct ectasia/hyperplasia and fibrocystic changes) in most (37.5%), followed by intra-ductal papilloma (32.5%), normal (16.6%), malignant (10.9%) and atypical (2.5%) changes in the rest. The presence of RBC in NC was 87% sensitive to detect any underlying pathology and 69% sensitive to predict malignancy at the expense of specificity. However, epithelial cells were found to be neither sensitive nor specific enough to detect underlying pathology or malignancy.

Conclusion: The aetiology remains benign in most patients presenting with ND in the absence of any underlying palpable or radiological abnormality. Even in patients without an underlying radiological abnormality, diagnostic
surgery should be offered if RBC is present in the nipple cytology.

Objective: Posterior uveal melanoma is the most common primary intraocular malignant tumor in adults. Despite the clinical significant treatment, still many patients die because of unpredictable metastases. It is difficult to predict clinical outcome in individual cases of ocular melanoma basing only on the intraocular size, because of the spectrum of clinical, morphologic, and cytologic changes and lack of discrete stages.

Design: The aim of the study was to evaluate CD3, CD20 and CD34 expression within choroidal melanoma in
correlation with clinic and pathological features. Participants: 35 cases of choroidal melanoma were evaluated.

Methods: Immunohistochemical reaction was used to evaluate CD3 and CD20 (T and B cells markers) and
CD34 (endothelial cells marker) expression, with positive and negative controls. Obtained results were estimated under the light microscope.

Results: T lymphocytes were present in 94.3% choroidal melanoma specimens, the most significant CD3
expression was noticed in choroidal melanoma in pT3 stage. Presence of B lymphocytes was observed only in 22.8% cases, but only in advanced tumors (pT3). CD34 expression was observed in 77.1% choroidal melanomas. A statistically significant correlation was observed between CD34 expression and staging- 84.2% choroidal melanomas in pT3 stage were positive for CD34 expression.

Conclusions: T cell infiltration is present in both low and high advanced ocular melanomas. B cells and thin wall vessels are present mainly in advanced choroidal melanoma and due to may be associated with tumor progression.

Introduction: Hirschsprung’s disease (HD) is rare (1/5000 newborns) congenital condition of the bowel nervous system, clinically characterized by dilatation and hypertrophy of colon. There are an increased number of mast cells (MC) in aganglionic bowel segments and in those with intestinal neuronal dysplasia (IND). The exact role of MC in HD is still not known. The aim of this study was to examine the colonic distribution of mast cells (MC) and the relationship between MC and nerve fibers and to estimate connections of these cells with gut nervous system. Material and Methods: Distal colon wall specimens were harvested from patients with normal bowel function and with normal ganglion cells, patients with constipations and with normal ganglion cells and a group of patients with Hirschsprung’s disease. The age of the patients ranged from 2nd day of live to 12 years. The density of the mast cells was calculated using a light microscope with digital camera. The connections of localization of a MC with nerve fibers and ganglions were taken under consideration and observed under light microscope. Test for the significance of differences between groups were performed using the Kruskal-Wallis test. Results: We revealed that there is a greater number of the MC in mucosa, lamina propria, submucosa and muscle layer of large bowel in patients with HD. There was no difference in density of the MC in serosa. Some of the colon MC in HD was located near or they were even attached to the hypertrophic nerve trunks. We found a correlation of MC number with the number of nerve fibers in HD patients. Conclusions: Higher number of MC and their localization in close connection with hypertrophied nerve trunks in patients with HD than in patients with constipation or normal bowel function, is a proof of the influence of MC on defected nervous system in aganglionic bowel. MC probably affect hypertrophied nerve trunks. Mobilization and activation of the mast cells which produce and store NGF can be directed to repair defective nervous system. Obviously, MC repairing reaction is not effective in restoring nervous system function of the gut.