Mean Number Of Positive Cells Research Articles

Quantitative assessment Cyclooxygenase – 2 immunohistochemical expression in different grades of oral epithelial dysplasia

Oral leukoplakia (OLP) is one the most common potentially malignant disorders account 0.2 to 4.9% of malignant transformation and dysplastic lesions vary from 3-6% this rate of malignant conversion depends on the different grades of epithelial dysplasia. Recent literature suggests that increased levels of COX-2 in potentially malignant disorders and malignant tumors show prominent genetic evidence in tumorigenesis. Assessment of Cyclooxygenase-2 – 2 (COX-2) immunohistochemical expression in different grades of oral epithelial dysplasia. Materials and Methodology A total of forty formalin-fixed, paraffin-embedded tissue blocks were obtained from departmental archives which are histopathological diagnoses of different grades of oral epithelial dysplasia each grade ten samples was included. Stained with COX-2 and positive cells were counted and tabulated for statistical analysis. Results Comparison between genders in oral leukoplakia males was more when compared with females. The mean age in mild epithelial dysplasia was more when compared with moderate and severe epithelial dysplasia. Expression of COX-2 was increased from mild to severe epithelial dysplasia increases. The mean number of positive cells was increased. A comparison of all groups was not statistically significant (p≥0.05). Conclusion: Oral leukoplakia (OLP) is one the most common potentially malignant disorders which accounts for 0.2 to 4.9% of malignant transformation. COX-2 can be used for early detection and prognostic makers in different grades of oral epithelial dysplasia. Keywords: Cyclooxygenase, Expression, Oral leukoplakia, Potentially malignant transformation

Diaph3 underlines tumor cell heterogeneity in glioblastoma with implications for treatment modalities resistance

Glioblastoma (GBM) is the most aggressive central nervous system (CNS) tumor with astrocytic differentiation. The growth pattern of GBM mimics that of the precursor cell migration during the fetal development of the brain. Diaphanous homolog (Diaph3) has been established to play a role in both CNS maturation and cancer progression as it is required both for cell migration and division. Furthermore, Diaph3 has been shown to play a role in malignant disease progression through hyperactivation of the EGFR/MEK/ERK in loss of expression and its overexpression correlating to hyperactivity of the mTOR pathway, both of which are with a well-established role in GBM. Herein, we aimed at establishing the diagnostic role of Diaph3 immunohistochemistry expression patterns in GBM and their possible implications for molecular response to different therapies. The study utilized a retrospective nonclinical approach. Results of Diaph3 immunohistochemical expression were compared to healthy controls and reactive gliosis and statistically analyzed for correlation with neuroradiological tumor parameters and patient survival. Healthy controls showed individual weakly positive cells, while reactive gliosis controls showed a strong expression in astrocytic projections. GBM samples showed a heterogeneous positive reaction to Diaph3, mean number of positive cells 62.66%, median 61.5, range 12-96%. Areas of migrating cells showed a strong diffuse cytoplasmic reaction. Cells located in the tumor core and those in areas of submeningeal aggregation had no antibody expression. Statistical analysis revealed no correlation with tumor size or patient survival. The different expression pattern of Diaph3 in healthy controls, reactive gliosis and GBM shows promise as a clinical differentiating marker. Despite Diaph3 expression not correlating with survival and tumor size in GBM, there is an accumulating body of evidence that Diaph3 correlates with mTOR activity and can thus be used as a predictor for response to rapamycin and taxanes, clinical studies of which have shown promising, if mixed results in GBM.

Tissue Ki67 proliferative index expression and pathological changes in hemodialysis arteriovenous fistulae: Preliminary single-center results.

Arteriovenous fistula (AVF) for hemodialysis integrates outward remodeling with vessel wall thickening in response to drastic hemodynamic changes. Aim of this study is to determine the role of Ki67, a well-established proliferative marker, related to AVF, and its relationship with time-dependent histological morphologic changes. All patients were enrolled in 1 year and stratified in two groups: (A) pre-dialysis patients submitted to first AVF and (B) patients submitted to revision of AVF. Morphological changes: neo-angiogenesis (NAG), myointimal thickening (MIT), inflammatory infiltrate (IT), and aneurysmatic fistula degeneration (AD). The time of AVF creation was recorded. A biopsy of native vein in Group A and of arterialized vein in Group B was submitted to histological and immunohistochemical (IHC) analysis. IHC for Ki67 was automatically performed in all specimens. Ki67 immunoreactivity was assessed as the mean number of positive cells on several high-power fields, counted in the hot spots. A total of 138 patients were enrolled, 69 (50.0%) Group A and 69 (50.0%) Group B. No NAG or MIT were found in Group A. Seven (10.1%) Group A veins showed a mild MIT. Analyzing the Group B, a moderate-to-severe MIT was present in 35 (50.7%), IT in 19 (27.5%), NAG in 37 (53.6%); AD was present in 10 (14.5%). All AVF of Group B with the exception of one (1.4%) showed a positivity for Ki67, with a mean of 12.31 ± 13.79 positive cells/hot spot (range 0-65). Ki67-immunoreactive cells had a subendothelial localization in 23 (33.3%) cases, a myointimal localization in SMC in 35 (50.7%) cases. The number of positive cells was significantly correlated with subendothelial localization of Ki67 (p = 0.001) and with NA (p = 0.001). Native veins do not contain cycling cells. In contrast, vascular cell proliferation starts immediately after AVF creation and persists independently of the time the fistula is set up. The amount of proliferating cells is significantly associated with MIT and subendothelial localization of Ki67-immunoreactive cells, thus suggesting a role of Ki-67 index in predicting AVF failure.

EXPRESSION OF MESENCHYMAL STEM CELL MARKERS IN THE STROMA OF ODONTOGENIC CYSTS AND TUMORS

Objective To investigate the immunoexpression of stem cell markers (Nanog, SOX2, Oct4, CD34) in odontogenic cysts and tumors. CD34 was investigated as a marker for stromal fibroblast/fibrocyte cells (CD34+SFCs). CD34+SFCs were also investigated ultrastructurally. The physical aggregation of collagen fibers was examined by picrosirius red stain and polarized light. Methods Ten cases each of primary odontogenic keratocysts (OKC), recurrent OKC, dentigerous cysts, ameloblastoma, unicystic ameloblastoma, and odontogenic myxoma and 7 syndromic OKC (sOKC) were included. Results were expressed as the mean number of positive cells per field. Changes (%) in polarization colors of the collagen fibers were assessed pending 90° rotation of the microscope table. Significance was set at P Results All markers except Oct4 were expressed by stromal cells in all lesions. Expression of SOX2 but not Nanog was significantly higher in tumors than in cysts (P Conclusions Stromal cells in odontogenic cysts and tumors seem to contribute to their biological behavior.

CDK4/6 inhibition mitigates stem cell damage in a novel model for taxane‐induced alopecia

Taxanes are a leading cause of severe and often permanent chemotherapy‐induced alopecia. As the underlying pathobiology of taxane chemotherapy‐induced alopecia remains poorly understood, we investigated how paclitaxel and docetaxel damage human scalp hair follicles in a clinically relevant ex vivo organ culture model. Paclitaxel and docetaxel induced massive mitotic defects and apoptosis in transit amplifying hair matrix keratinocytes and within epithelial stem/progenitor cell‐rich outer root sheath compartments, including within Keratin 15+ cell populations, thus implicating direct damage to stem/progenitor cells as an explanation for the severity and permanence of taxane chemotherapy‐induced alopecia. Moreover, by administering the CDK4/6 inhibitor palbociclib, we show that transit amplifying and stem/progenitor cells can be protected from paclitaxel cytotoxicity through G1 arrest, without premature catagen induction and additional hair follicle damage. Thus, the current study elucidates the pathobiology of taxane chemotherapy‐induced alopecia, highlights the paramount importance of epithelial stem/progenitor cell‐protective therapy in taxane‐based oncotherapy, and provides preclinical proof‐of‐principle in a healthy human (mini‐) organ that G1 arrest therapy can limit taxane‐induced tissue damage.

Distribution of neuroendocrine marker-positive cells in colorectal cancer tissue and adjacent mucosa.

617 Background: Neuroendocrine carcinoma (NEC) is a rare disease and has been reported to most frequently arise in the right side of the colon. In the 2010 WHO classification, mixed adenoneuroendocrine carcinoma (MANEC) was defined as a neoplasm consisting of NEC and adenocarcinoma components. To clarify the histogenesis of NEC, we attempted to detect neuroendocrine marker-positive cells in cancer tissue and in the adjacent mucosa in patients with adenocarcinoma. Methods: The study group comprised 390 patients with Stage II or III colorectal adenocarcinoma between 2007 and 2012. Immunostaining was performed with anti chromogranin A, synaptophysin, and CD56 antibodies. Cases with positively stained cells in cancer tissue were defined as positive. In the adjacent mucosa, at least 5 cm from the tumor, the numbers of positive cells per 15 HPF were measured. Results: Tumor location was right side in 181 patients, left side in 173, and the rectum in 36 patients. Positive rates of Chromogranin A in cancer tissues were 23.7% in the right colon, 13.2% in the left colon, and 19.4% in the rectum. Those of synaptopysin were 35.3%, 21.9%, and 30.6%, respectively. Those of CD56 were 22.6%, 8.0%, and 16.7%, respectively. Positive rates of these three markers in right colon were significantly higher than those in left colon and rectum. (p = 0.0115, p = 0.0054, p = 0.0062). In the adjacent mucosa, the mean numbers of positive cells for chromogranin A were 62.2 ± 20.5 in the right colon, 131.9 ± 44.7 in the left colon, and 243.7 ± 60.2 in the rectum (p < 0.001). Those for synaptophysin were 47.7 ± 23.5, 95.3 ± 35.1, and 156.9 ± 56.8, respectively. (p < 0.001). There were no significant differences in the number of positive cells for CD56 among the sites (p = 0.295). Conclusions: In cancer tissue, the rate of positive staining for neuroendocrine marker-positive cells was higher in the right side of the colon, whereas in normal mucosa the rates of positive staining for these cells were higher in the sigmoid colon and the rectum. These results suggest that neuroendocrine marker-positive cells are an acquired characteristic of cancer tissue.

The effects of maternal diabetes and insulin treatment on neurogenesis in the developing hippocampus of male rats

Diabetes in pregnancy is associated with an increasing risk of congenital malformations and central nervous system disorders (CNS) especially hippocampal neuronal circuitry disruption as a discreet region involved in neurogenesis phenomenon. This study aimed to investigate the effect of maternal diabetes and insulin treatment on the expression and distribution pattern of NeuN and DCX as two important markers of neurogenesis paradigm in developing rat hippocampus. All animals were randomly divided into three groups as follows: Control group, Diabetic (STZ-D), Diabetic treated with insulin (STZ-INS). Diabetes was induced in Wistar female rats by Sterptozotocin intraperitoneal injection (single does). Following confirmation of diabetes, animals were mated with non-diabetic males. Four to six units of protamine-Zinc insulin were delivered subcutaneously (SC) in insulin treated group. At the post-natal day 14 (P14), the brain of male offspring’s were removed for further study. In fact Immunofluorescence staining and Real time − PCR assays are used for evaluation of neurogenesis phenomenon. Our results showed a significant higher level of hippocampal DCX expression and an increase in the mean number of DCX positive cells in the DG of diabetic group male offspring (P < 0.05). We also found an insignificant up-regulation in the expression of DCX and the mean number of positive cells in the insulin-treated diabetic group neonates as compared to control group (P > 0.05). Nevertheless the results of immunofluorescence staining for NeuN also indicated that the mean number of NeuN+ cells was significantly lower in dentate gyrus of diabetic group male offspring (P < 0.05). Besides, there were significant down- regulation in the hippocampal mRNA expression of NeuN in diabetic pups compare to control (P < 0.05 each). Our results revealed that diabetes during pregnancy has an adverse effect on the hippocampal neurogenesis in rat neonates. Furthermore, the control of glycemia by insulin is sufficient to prevent the alterations in expression of neurogenesis markers.

A comparative study of odontogenic keratocyst and orthokeratinized odontogenic cyst using Ki67 and α smooth muscle actin.

Aim:This study aimed to demonstrate and evaluate the expression of stromal myofibroblasts (MFs) and epithelial cell proliferation using α-smooth muscle actin (α-SMA) and Ki67 markers, respectively, in odontogenic keratocyst (OKC) and orthokeratinized odontogenic cyst (OOC) to correlate their aggressive behavior.Materials and Methods:Twenty cases of OKC and twenty cases of OOC were stained with α-SMA and Ki67 markers for demonstration of stromal MFs and epithelial cell proliferation, respectively, and ten cases of well-differentiated squamous cell carcinoma were used as positive control. Assessment of the number of α-SMA-positive stromal cells and Ki67-positive epithelial cells determined by MFs and proliferative epithelial cell frequency in 10 high-power fields (×400) was presented as the mean number of positive cells per field.Statistical Analysis:Kruskal–Wallis and Mann–Whitney test were used to analyze the difference in the mean number of α-SMA- and Ki67-positive cells per field between OKC and OOC.Results:The mean number of positively stained cells for α-SMA and Ki67 is significantly higher in OKC compared to OOC.Conclusion:Impression is that, the different behaviors of these two entities are compatible with their immunohistochemical view. The high value of stromal MFs and proliferative epithelial cells in OKC in comparison to OOC indicates its aggressiveness and potential for recurrence.

Beyond Histological Remission: Intramucosal Calprotectin as a Potential Predictor of Outcomes in Ulcerative Colitis.

Histological remission and low faecal calprotectin are positive prognostic factors in ulcerative colitis [UC]. Intramucosal calprotectin [iMC], which can be readily determined by immunohistochemistry, has not so far been evaluated as a predictor of outcome in UC. We aimed to investigate the relationship between iMC and clinical, endoscopic, and histological measures of remission in UC, and the independent prognostic value of iMC. Ambulant patients with UC were recruited for a study comparing clinical activity indices. Sigmoidoscopy and biopsy were performed at the index visit. Clinical, endoscopic, and histological activity were scored and iMC semi-quantitatively measured using immunohistochemistry for the S100A8/9 heterodimer on colonic biopsies, scored as the mean number of positive cells in five high-power fields [HPF]. At the end of follow-up [6 years], data on steroid use, hospitalisation, and colectomy ['adverse outcomes'] were collected. iMC was determined in 83 patients and 20 controls, and correlated with clinical, endoscopic, and histological activity [r = 0.51, 0.65, 0.53, p > 0.001, respectively]. iMC was lowest (median 2.4, interquartile range [IQR]: 5.2-5, p < 0.001) in patients with concordance between clinical, endoscopic, and histological remission. Median iMC > 5/HPF was associated with adverse outcome (hazard ratio [HR] 3.36, confidence interval [CI] 1.58, 7.15, p < 0.001). Only 53%, 33%, and 25% of patients in histological remission with iMC > 5 cells/HPF avoided an adverse outcome after 1, 3, and 6 years, respectively. iMC was lowest in patients with concordant clinical, endoscopic, and histological remission. Median iMC > 5/HPF was associated with adverse outcomes despite histological remission. Therefore iMC is a potentially useful independent marker of activity.

Immunohistochemistry Study on Androgen and Estrogen Receptors of Rat Seminal Vesicle Submitted to Simultaneous Alcohol-Nicotine Treatment.

Alcohol consumption is habitually accompanied by the use of other psychoactive substances, mostly tobacco. Nicotine and alcohol affect male accessory reproductive glands function. Most studies have been done on pathologic features of prostate, but there has been no systematic study on the seminal vesicle. Therefore, the aim of current study was to investigate the distribution of androgen receptor (AR) and estrogen receptors-beta (ER-β) immune reactivities following long-term treatment of alcohol, nicotine or a combination of both substances. In this experimental study, a total of 40 adult Wistar rats, nine weeks of age, were used. Animals were randomly divided into four groups, including: i. Control group receiving normal saline 0.09%, ii. Ethanol group receiving ethanol 20% (2 ml/kg, via gavage), iii. Nicotine group receiving nicotine (0.1 mg/kg, subcutaneous injection), and iv. Ethanol-nicotine group receiving simultaneous ethanol 20% (2 ml/kg) and nicotine (0.1 mg/kg) treatment. All treatment lasted for eight weeks. Prior to intracardiac perfusion, blood sample was collected from left ventricle. The seminal vesicles were isolated and processed for paraffin blocking. The sample tissues were then studied for distribution of AR and ER-β immunereactivities using immunohistochemical (IHC) staining method. One way analysis of variance (ANOVA) and Tukey's test were performed for data analysis. A value of P<0.05 was considered significant. Our results revealed that the lowest mean number of positive cells belonged to the animals of ethanol-nicotine group that was followed by the ethanol, nicotine, and control groups, respectively. However, there was no significant difference regarding serum testosterone level among experimental groups. It was concluded that combination of both ethanol and nicotine may be a crucial factor in the expression levels of AR and ER-β.

Overexpression of MutSα Complex Proteins Predicts Poor Prognosis in Oral Squamous Cell Carcinoma.

The DNA mismatch repair (MMR) system is responsible for the detection and correction of errors created during DNA replication, thereby avoiding the incorporation of mutations in dividing cells. The prognostic value of alterations in MMR system has not previously been analyzed in oral squamous cell carcinoma (OSCC).The study comprised 115 cases of OSCC diagnosed between 1996 and 2010. The specimens collected were constructed into tissue microarray blocks. Immunohistochemical staining for MutSα complex proteins hMSH2 and hMSH6 was performed. The slides were subsequently scanned into high-resolution images, and nuclear staining of hMSH2 and hMSH6 was analyzed using the Nuclear V9 algorithm. Univariable and multivariable Cox proportional hazard regression models were performed to evaluate the prognostic value of hMSH2 and hMSH6 in OSCC.All cases in the present cohort were positive for hMSH2 and hMSH6 and a direct correlation was found between the expression of the proteins (P < 0.05). The mean number of positive cells for hMSH2 and hMSH6 was 64.44 ± 15.21 and 31.46 ± 22.38, respectively. These values were used as cutoff points to determine high protein expression. Cases with high expression of both proteins simultaneously were classified as having high MutSα complex expression. In the multivariable analysis, high expression of the MutSα complex was an independent prognostic factor for poor overall survival (hazard ratio: 2.75, P = 0.02).This study provides a first insight of the prognostic value of alterations in MMR system in OSCC. We found that MutSα complex may constitute a molecular marker for the poor prognosis of OSCC.

Altered expression of p53, but not Rb, is involved in canine prostatic carcinogenesis

Abnormalities in the retinoblastoma (Rb) and p53 tumour suppressor gene have been frequently detected in human and canine cancers, but never investigated in canine prostate cancer, considered a good model for the advanced and aggressive androgen-resistant prostate cancer in men. Therefore, the aim of this study was to evaluate the immunohistochemical expression of Rb and p53 in 6 normal canine prostates, 15 canine prostates with benign prostatic hyperplasia (BPH) and 10 prostatic carcinomas (PCs). In all normal samples, p53 was expressed in low number of epithelial cells, while a greater number of positive cells were observed in BPH and PC. The mean number of positive cells was statistically significantly higher in PCs than normal and hyperplastic prostates. A cytoplasmic or nucleo-cytoplasmic staining was observed in 5 out of 10 PCs. Rb protein was expressed in high number of normal, hyperplastic and neoplastic cells without a statistically significant differences. Considering that Rb is frequently lost in human prostate cancer, we suggest that Rb is not involved in canine prostatic carcinogenesis. On the other hand, the increased expression of p53 that corresponds to genetic defects in the p53 gene may be associated with the malignant growth of canine prostate cancer, conferring an apoptosis-resistant phenotype.

Effect of maternal diabetes on gliogensis in neonatal rat hippocampus.

Background:Diabetes in pregnancy is a common metabolic disorder associated with various adverse outcomes in the offspring including impairments in attention and memory and alterations in social behavior. Glial cells are proven to have a critical role in normal function of neurons, and alteration in their activity could contribute to disturbance in the brain function. The aim of this study was to investigate the effect of maternal diabetes on hippocampal mRNA expression and distribution pattern of glial fibrillary acidic protein (GFAP) immunoreactive glial cells in the dentate gyrus (DG) of rat neonate at postnatal day 14 (P14).Materials and Methods:Wistar female rats were randomly allocated in control, diabetic, and insulin-treated diabetic groups. Diabetes was induced by injection of streptozotocin from 4 weeks before gestation until parturition. After delivery, the male offspring was euthanized at P14.Results:Our results showed a significant higher level of hippocampal GFAP expression and an increase in the mean number of GFAP positive cells in the DG of diabetic group offspring (P < 0.05). We also found an insignificant up-regulation in the expression of GFAP and the mean number of positive cells in the insulin-treated diabetic group neonates as compared to control group (P > 0.05).Conclusion:The present study revealed that diabetes during pregnancy strongly increased the glial cells production in the developing rat hippocampus.

Immunohistochemical evaluation and biological role of stromal myofibroblasts in odontogenic keratocyst, dentigerous cyst, and ameloblastoma: A comparative study

Context: Stromal myofibroblasts (MFs) are key cells for connective tissue remodeling and interact with epithelial cells and other connective tissue cells to control phenomena as tumor invasion and angiogenesis thereby contributing to their biological behavior. Aims: The study assesses the frequency of stromal MF in solid ameloblastoma (SA), odontogenic keratocyst (OKC), and dentigerous cyst (DC) and relates it to their biological behavior. Settings and Design: Ten cases of each lesion were taken and stained immunohistochemically for alpha-smooth muscle actin (α-SMA) which is a marker for stromal MF. Materials and Methods: Ten cases each of SA, OKC, and DC were included in the study. Cases of oral squamous cell carcinoma (SCC, n = 5) served as the baseline for comparison as they are aggressive lesions expressing increased number of stromal MFs. The frequency of MFs was assessed as the number of α-SMA-positive stromal cells in 10 high-power fields and presented as the mean number of positive cells per field. Statistical Analysis Used: Differences in the mean number of α-SMA-positive cell per field among SA, OKC, DC, and SCC were analyzed using one-way ANOVA test. Results: Counts showed that mean number of α-SMA-positive MFs in SA, OKC, and DC were 24.56 (±4.63), 21.37 (±4.17), and 8.03 (±2.15), respectively. Results showed that the mean number of stromal MFs in SA and OKC was significantly higher than that in DC (8.03 ± 2.15) (P 0.05). Conclusion: Activated MF participates in the matrix degradation process which is considered to be one of the main forces in tumor growth and invasion. Among odontogenic lesions, ameloblastoma and OKC (presently termed as keratocystic odontogenic tumor) are well known for their higher growth and recurrence potential. They tend to show burrowing growth pattern. Various studies have evaluated the epithelial factors responsible for their growth potential; we in our study have tried to relate the emergence of stromal MF to the biological behavior of these lesions. The frequency of stromal MF in OKC and ameloblastoma was almost similar to that in SCC, thereby implying that MF can contribute to the biological behavior of these odontogenic lesions.

Immunohistochemical Expression of Angiogenic Factors by Neoplastic Epithelial Cells Is Associated With Canine Prostatic Carcinogenesis

The dog is the only species, other than humans, in which spontaneous prostatic cancer occurs; therefore, dogs are a valuable model for the study of factors that regulate tumor progression. Angiogenesis is important in the development and spread of a variety of cancers, including prostate cancer. To better define the role of cancer epithelial cells in prostate cancer neovascularization, immunohistochemical staining for angiogenic factors (vascular endothelial growth factor [VEGF], platelet endothelial cell adhesion molecule-1 [PECAM-1], Tie-2, and fibroblast growth factor-2 [FGF-2]) was performed on formalin-fixed, paraffin-embedded tissues from 10 normal prostates, 15 hyperplastic prostates, and 11 prostatic carcinomas from dogs. Normal and hyperplastic epithelial cells were negative for PECAM-1, VEGF, and Tie-2, while the same markers were expressed with a variable intensity of cytoplasmic staining by neoplastic cells. Mild to moderate FGF-2 staining was detected in all normal prostates with less than 10% of positive cells, mainly distributed in the basal layer. The percentage of FGF-2-positive hyperplastic cells was variable, with both basal and secretory cells exhibiting a perinuclear to diffuse cytoplasmic staining. The mean number of positive cells and the intensity of staining were higher in prostatic carcinomas than normal and hyperplastic prostates. Moreover, microvessel density analyzed on PECAM-1-stained slides was increased in prostate cancer compared with normal and hyperplastic prostates. Therefore, prostatic neoplastic cells are capable of simultaneous expression of various angiogenic factors and may increase tumor proliferation and angiogenesis in a paracrine and autocrine fashion.

Immunoexpression of HIF-1 a in Oral Vascular Lesions

To understand the pathogenesis of oral vascular lesions, we investigated hypoxia-inducible factor-1α (HIF-1α) immunoexpression in hemangioma and pyogenic granuloma (PG). Study Design: The sample consisted of 30 cases of hemangiomas and 30 of PG. The first diagnoses were reclassified based on expression of human glucose transporter (GLUT-1). Subsequently, lesions were immunohistochemically evaluated for the expression of HIF-1α. Positive and negative cells were counted, and the mean number of positive cells was calculated for each case.

Study of the etiopathogenesis and differential diagnosis of oral vascular lesions by immunoexpression of GLUT‐1 and HIF‐1α

To investigate whether the immunohistochemical expression of GLUT-1 and HIF-1α is related to the diagnosis and pathogenesis of oral vascular lesions. Thirty cases each of pyogenic granuloma (PG) and hemangioma were studied. Antibodies against GLUT-1 and HIF-1α were detected by immunoperoxidase staining in 3-μm histological sections, and the results were analyzed qualitatively and quantitatively, respectively. Positive and negative cells were counted, and the mean number of positive cells was calculated for each case. The initial diagnosis of hemangioma was maintained in only 7 (23%) of the 30 cases studied, which were positive for GLUT-1. The remaining 23 cases were reclassified as vascular malformation (VM) (n = 13) and PG (n = 10) due to the absence of staining. The endothelium of blood vessels was negative for GLUT-1 in all cases initially diagnosed as PG (n = 30). The percentage of HIF-1α-positive cells was higher in cases of PG, followed by hemangiomas and VMs (P = 0.005). Histological features are not sufficient to establish the correct diagnosis of oral hemangiomas, and an accurate anamnesis is essential in these cases. In addition, these findings demonstrate that vascular lesions express mediators of angiogenesis, HIF-1α, and suggest that his process may play a role in the pathogenesis of vascular.

Evaluation of stromal myofibroblasts expression in keratocystic odontogenic tumor and orthokeratinized odontogenic cysts: A comparative study

Objective:Keratocystic odontogenic tumor (KCOT) has an aggressive clinical course and a high tendency of recurrence, while orthokeratinized odontogenic cyst (OOC) has different characteristics and does not show aggressive behaviour. Even the treatment of these two lesions varies considerably. A large number of epithelial molecules have been studied in order to differentiate odontogenic keratocyst (OKC) from OOC, but stromal factors have not been adequately studied. Recently, tumor stroma has evolved as a particular field of interest. In the present study, we aim to evaluate and compare the expression of stromal myofibroblasts (MFs) in these entities and correlate it to its aggressive behavior. The term ‘keratocystic odontogenic tumor’ has been introduced by WHO in 2005 for odontogenic keratocyst keeping in mind its aggressive behavior, however still many pathologists and clinicians use the term OKC synonymously.Materials and Methods:A total of 10 cases of KCOT and 10 cases of OOC were stained for alpha-smooth muscle actin (αSMA) for demonstration of stromal MFs. MF frequency was assessed as the number of αSMA-positive stromal cells in 10 high power fields, presented as the mean number of positive cells per field.Results:Counts showed that the mean number of positive cells in KCOT (20.6 ± 2.05) was significantly higher than that seen in OOC (10.4 ± 1.06) (P < 0.05).Conclusion:The different behaviors of these lesions are compatible with the finding of the present study. The increased number of stromal MFs in KCOT in comparison to OOC correlates with its aggressive behavior and increased tendency towards recurrence.

Toll-Like Receptors in Idiopathic Orbital Inflammation

A prior investigation has demonstrated that innate immune-specific cytokines are enriched in idiopathic orbital inflammation (IOI). To further document the role of innate immunity in IOI, the authors sought to determine whether toll-like receptors (TLRs) are present in biopsy specimens of this disorder. Immunohistochemical staining for TLR2, TLR3, and TLR4 was performed on biopsy specimens taken from patients with IOI, and the number of TLR-positive cells was counted across five 40× light microscopic fields. These results were compared with an isotype control and with orbital adipose tissue taken from patients without evidence of inflammation. All IOI specimens demonstrated positivity for all 3 TLRs, and sections stained for isotype controls did not demonstrate any positivity. Furthermore, orbital adipose tissue did not demonstrate any significant signal. The mean number of positive cells was 24.4 cells/high power field (hpf; standard deviation = 11.6 cells/hpf), 7.23 cells/hpf (standard deviation = 5.59 cells/hpf), and 11.7 cells/hpf for TLR2, TLR3, and TLR4, respectively. This study provides the first documentation of TLRs in orbital disease. Toll-like receptors are present in IOI, and IOI may represent an aberrant innate immune response. Interference with TLRs may represent an additional potential therapeutic mechanism in the management of IOI.

Comparative Susceptibility of Some Avian and Mammalian Lymphoblastoid Cell Lines to Chicken Anemia Virus

Although MSB1 is the most-commonly used lymphoblastoid cell line for isolation of chicken anemia virus (CAV), some researchers have reported a few biological drawbacks. None of them were supported by the results of the present study. Another four avian (HP1, HP2, BK3 and CU10) and two mammalian (BTL-26 and KO-1) cell lines were investigated for susceptibility to the TK-5803 and AH-9409 strains. Both strains caused CPE on BK3 like MSB1. The mean number of positive cells for each strain in MSB1 and BK3 were not significantly different. The majority of the HP2, CU10 and HP1 cells showed no CPE. The virus titers of both strains were higher in MSB1 and BK3 (10(6.5-7.5) TCID(50)/0.1 ml) than in HP2, CU10 and HP1 (10(3.5-4.5) TCID(50)/0.1 ml). BTL-26 and KO-1 were resistant to CAV. BK3 could be used for isolation of CAV.