Browsing by Author "Milenkovic, Sanja (57220419015)"

Purpose: Despite major advances in the treatment of diffuse large B cell lymphoma (DLBCL), approximately one third of the patients progress or die, suggesting the existence of additional oncogenic events. The purpose of this study was to evaluate the prognostic value of the "Hans classifier", and BCL2 and MYC protein expression and gene alterations in DLBCL patients treated with CHOP or R-CHOP chemotherapy over a 5-year period. Furthermore, we tried to correlate these parameters with the International Prognostic Index (IPI). Methods: The immunohistochemical (IHC) expression of CD10, BCL6, MUM1 and BCL2 on paraffin-embedded formalin-fixed tumor samples from 103 centroblastic DLBCLs was analyzed. IHC expression of MYC and fluorescence in situ hybridization (FISH) for MYC and BCL2 gene alterations was performed on 67 samples using the tissue microarray (TMA) method. Results: The Hans algorithm was not predictive of survival in both therapy groups. No significant difference in BCL2 and MYC alterations or MYC protein expression in relation to complete response (CR), event-free survival (EFS) and overall survival (OS) was observed in our study. High IPI correlated significantly with poor outcome and it was identified as independent prognostic factor for OS and EFS (both p=0.000). The 5-year OS was 61% in the R-CHOP compared to 38% in the CHOP group (p=0.007). Rituximab significantly improved the OS in the BCL2 positive (60 vs 29%, p=0.008), and the BCL6 negative (73 vs 25%, p=0.001) cases. Conclusion: IPI is an independent prognosticator for DLBCL patients and the addition of rituximab significantly improved survival. Furthermore, patients with BCL2+ and BCL6- DLBCL benefited from R-CHOP.

Purpose: Despite major advances in the treatment of diffuse large B cell lymphoma (DLBCL), approximately one third of the patients progress or die, suggesting the existence of additional oncogenic events. The purpose of this study was to evaluate the prognostic value of the "Hans classifier", and BCL2 and MYC protein expression and gene alterations in DLBCL patients treated with CHOP or R-CHOP chemotherapy over a 5-year period. Furthermore, we tried to correlate these parameters with the International Prognostic Index (IPI). Methods: The immunohistochemical (IHC) expression of CD10, BCL6, MUM1 and BCL2 on paraffin-embedded formalin-fixed tumor samples from 103 centroblastic DLBCLs was analyzed. IHC expression of MYC and fluorescence in situ hybridization (FISH) for MYC and BCL2 gene alterations was performed on 67 samples using the tissue microarray (TMA) method. Results: The Hans algorithm was not predictive of survival in both therapy groups. No significant difference in BCL2 and MYC alterations or MYC protein expression in relation to complete response (CR), event-free survival (EFS) and overall survival (OS) was observed in our study. High IPI correlated significantly with poor outcome and it was identified as independent prognostic factor for OS and EFS (both p=0.000). The 5-year OS was 61% in the R-CHOP compared to 38% in the CHOP group (p=0.007). Rituximab significantly improved the OS in the BCL2 positive (60 vs 29%, p=0.008), and the BCL6 negative (73 vs 25%, p=0.001) cases. Conclusion: IPI is an independent prognosticator for DLBCL patients and the addition of rituximab significantly improved survival. Furthermore, patients with BCL2+ and BCL6- DLBCL benefited from R-CHOP.

Summary: Aim: To evaluate contact telescopy findings for estimation of blood vessel changes in vocal fold mucosa in patients with Reinke's edema. Histological features significant for diagnosis of microvascular vocal fold alteration were correlated with clinical findings. Methods: In 80 patients with Reinke's edema, laryngoscopy and video-telescopy image analysis of vocal folds were performed. Vocal fold mucosa biopsies were histologically analyzed and compared with contact telescopy findings. An interesting aspect of vocal fold microcirculation found both by contact telescopy imiging and by histological specimens was described. Results: Contact telescopy in vivo revealed different forms of pathological blood vessel networks with unusual appearance of loops or branching. Some dilated varicose vascular channels had very thin walls, and within atypical capillaries, partial erythrocyte accumulation was found. Details of blood flow are also visible, showing multidirectional and discontinuous blood flow in neighboring vessels. Conclusion: The noninvasive contact telescope technique is very useful as an additional diagnostic tool for defining a condition of a subepithelial Reinke's space in a very short period of time. The great advantage of contact telescopy is systematic in vivo and in situ observation of microvascular details in the vocal folds. The contact technique allows dynamic follow-up of the microcirculation in Reinke's edema as well as simultaneous consultation of a pathologist in the operating theater. © 2007 The Voice Foundation.

Summary: Aim: To evaluate contact telescopy findings for estimation of blood vessel changes in vocal fold mucosa in patients with Reinke's edema. Histological features significant for diagnosis of microvascular vocal fold alteration were correlated with clinical findings. Methods: In 80 patients with Reinke's edema, laryngoscopy and video-telescopy image analysis of vocal folds were performed. Vocal fold mucosa biopsies were histologically analyzed and compared with contact telescopy findings. An interesting aspect of vocal fold microcirculation found both by contact telescopy imiging and by histological specimens was described. Results: Contact telescopy in vivo revealed different forms of pathological blood vessel networks with unusual appearance of loops or branching. Some dilated varicose vascular channels had very thin walls, and within atypical capillaries, partial erythrocyte accumulation was found. Details of blood flow are also visible, showing multidirectional and discontinuous blood flow in neighboring vessels. Conclusion: The noninvasive contact telescope technique is very useful as an additional diagnostic tool for defining a condition of a subepithelial Reinke's space in a very short period of time. The great advantage of contact telescopy is systematic in vivo and in situ observation of microvascular details in the vocal folds. The contact technique allows dynamic follow-up of the microcirculation in Reinke's edema as well as simultaneous consultation of a pathologist in the operating theater. © 2007 The Voice Foundation.

Purpose: The purpose of this study was to examine wheth er cytomegalovirus (CMV) is present in different histologi cal types of salivary gland cancer (SGC) by detecting CMV immediate-early (IE) and early gene products, and to deter mine the presence of its association with the overexpression of interleukin (IL)-6. Methods: Immunohistochemical analysis of 92 cases of dif ferent histological types of SGC was performed to determine the presence of IL-6 and CMV antigen and its intensity in tumor tissue. Twenty samples of normal salivary gland tis sue obtained during autopsy served as healthy controls. Results: CMV antigens were not found in healthy acinar tissue of salivary glands, but were expressed in epithelium of salivary gland ducts. Negative expression of CMV an tigens was also found in salivary gland tissue surround ing tumors. On the other hand, CMV was detected in 65/92 SGC cases (70.6%). Higher expression of IL-6 was found in SGC (70.7%) than in normal tissue (20%). There was a high association of CMV antigen presence with the presence of IL-6, and with the IL-6 expression intensity. Conclusions: Positive expression of CMV antigens in a high percentage of SGC cells suggests that it might play an important role in carcinogenesis by increasing IL-6 pro duction and leading to inhibition of apoptosis and tumor development.

Purpose: The purpose of this study was to examine wheth er cytomegalovirus (CMV) is present in different histologi cal types of salivary gland cancer (SGC) by detecting CMV immediate-early (IE) and early gene products, and to deter mine the presence of its association with the overexpression of interleukin (IL)-6. Methods: Immunohistochemical analysis of 92 cases of dif ferent histological types of SGC was performed to determine the presence of IL-6 and CMV antigen and its intensity in tumor tissue. Twenty samples of normal salivary gland tis sue obtained during autopsy served as healthy controls. Results: CMV antigens were not found in healthy acinar tissue of salivary glands, but were expressed in epithelium of salivary gland ducts. Negative expression of CMV an tigens was also found in salivary gland tissue surround ing tumors. On the other hand, CMV was detected in 65/92 SGC cases (70.6%). Higher expression of IL-6 was found in SGC (70.7%) than in normal tissue (20%). There was a high association of CMV antigen presence with the presence of IL-6, and with the IL-6 expression intensity. Conclusions: Positive expression of CMV antigens in a high percentage of SGC cells suggests that it might play an important role in carcinogenesis by increasing IL-6 pro duction and leading to inhibition of apoptosis and tumor development.

Epithelial to mesenchymal transition (EMT) is a feature of several types of human cancer, including oral squamous cell carcinoma (OSCC). In the present study, tumor and margin cell cultures obtained from patients with OSCC were used to determine the expression patterns of certain EMT-associatedmarkers,includingvimentin,α-smoothmuscle actin, SLUG and SNAIL. In addition, other EMT-associated features, including clonal, proliferative and migratory potential were compared between the two cell types. Cell cultures were generated from tumor and margin tissue samples from 6 patients and cultured up to the fifth passage. EMT marker expression was assessed by reverse transcription-quantitative PCR. Cell proliferation, colony formation and scratch wound healing assays were conducted to characterize the two cell types in terms of proliferation rates, clonality and motility. All of the studied markers were expressed in tumor and margin cells. Although no significant differences were noted with regard to the aforementioned markers, their expression tended to be higher in margin cultures than in tumor cultures. The expressions of the EMT markers were also higher in the fifth passage compared with those noted at the first with a few exceptions. The rates of proliferation and cell migration were decreased during passages, while the number of colonies was increased in both types of cell culture. Tumor and margin cells indicated certain similarities with regard to EMT transition characteristics. © 2020 Spandidos Publications. All rights reserved.

Epithelial to mesenchymal transition (EMT) is a feature of several types of human cancer, including oral squamous cell carcinoma (OSCC). In the present study, tumor and margin cell cultures obtained from patients with OSCC were used to determine the expression patterns of certain EMT-associatedmarkers,includingvimentin,α-smoothmuscle actin, SLUG and SNAIL. In addition, other EMT-associated features, including clonal, proliferative and migratory potential were compared between the two cell types. Cell cultures were generated from tumor and margin tissue samples from 6 patients and cultured up to the fifth passage. EMT marker expression was assessed by reverse transcription-quantitative PCR. Cell proliferation, colony formation and scratch wound healing assays were conducted to characterize the two cell types in terms of proliferation rates, clonality and motility. All of the studied markers were expressed in tumor and margin cells. Although no significant differences were noted with regard to the aforementioned markers, their expression tended to be higher in margin cultures than in tumor cultures. The expressions of the EMT markers were also higher in the fifth passage compared with those noted at the first with a few exceptions. The rates of proliferation and cell migration were decreased during passages, while the number of colonies was increased in both types of cell culture. Tumor and margin cells indicated certain similarities with regard to EMT transition characteristics. © 2020 Spandidos Publications. All rights reserved.

Heat shock protein family B (small) member 8 (HSPB8) promotes chaperone-assisted selective autophagy (CASA), which assures proteostasis in muscles and neurons. HSPB8 frameshift mutations found in neuromyopathies are translated on the same frame, generating the same C-terminal extension, which causes HSPB8 aggregation and proteostasis defects. Here, we describe three novel HSPB8 frameshift variants, translated to protein using the third alternative frame to stop codons downstream to the canonical one and to the one used by other known HSPB8 frameshift mutants. Therefore, these variants are predicted to encode a C-terminal extension that is different in length and amino acids. HSPB8 c.562delC and c.520_523delTACT were identified in two unrelated sporadic patients, while c.515delC, in a familial case of early-onset myopathy. Patients may differentially exhibit additional pathological features, such as neuropathy, respiratory insufficiency, and, remarkably, severe cardiomyopathy. Skeletal muscle biopsies revealed variations in fiber size, atrophy, multiple vacuoles, fat infiltration, and eosinophilic inclusions. In a reconstituted cell model of disease the expression of one representative novel HSPB8 mutant results in i) aggregation of the HSPB8 mutant, ii) sequestration of both the HSPB8 wild-type and CASA complex members, as well as iii) the autophagy receptor sequestosome-1 (SQSTM1/p62), iv) accumulation of ubiquitinated substrates, and v) defects in CASA-mediated degradation. Our results prove that the last exon of the HSPB8 gene is highly susceptible to pathogenic mutations, resulting in a wider phenotypic spectrum associated with HSPB8 frameshift variants. Our studies suggest the importance of HSPB8 genetic testing not only for neuropathy and myopathy but also for cardiomyopathy. © The Author(s) 2025.

Limb-girdle muscular dystrophy (LGMD) type 2A (calpainopathy) is an autosomal recessive disease caused by mutation in the CAPN3 gene. The aim of this study was to examine genetic and phenotypic features of Serbian patients with calpainopathy. The study comprised 19 patients with genetically confirmed calpainopathy diagnosed at the Neurology Clinic, Clinical Center of Serbia and the Clinic for Neurology and Psychiatry for Children and Youth in Belgrade, Serbia during a ten-year period. Eighteen patients in this cohort had c.550delA mutation, with nine of them being homozygous. In majority of the patients, disease started in childhood or early adulthood. The disease affected shoulder girdle - upper arm and pelvic girdle - thigh muscles with similar frequency, with muscles of lower extremities being more severely impaired. Facial and bulbar muscles were spared. All patients in this cohort, except two, remained ambulant. None of the patients had cardiomyopathy, while 21% showed mild conduction defects. Respiratory function was mildly impaired in 21% of patients. Standard muscle histopathology showed myopathic and dystrophic pattern. In conclusion, the majority of Serbian LGMD2A patients have the same mutation and similar phenotype. © Gaetano Conte Academy - Mediterranean Society of Myology.

Purpose: Since several studies have proposed that epithelial ovarian cancer should not be considered as a single disease entity and that it results from an accumulation of genetic changes, we aimed to assess the polymorphic expression of major cytosolic glutathione S-transferases (GSTM1, T1, A1 and P1) with respect to ovarian cancer susceptibility and aggressiveness. Methods: This case-control study was conducted on 93 newly diagnosed epithelial ovarian cancer patients and 178 healthy matched controls. The multiplex polymerase chain reaction (PCR) was used to detect homozygous deletions ofGSTMl and GSTT1 genes. Analysis of the single nucleotide polymorphism (SNP) GSTA1 C69T was performed using PCR-restrictionfragment length polymorphism (RFLP), while for SNP GSTP1 Ile105Val real-time PCR was used. Results: No significant association to ovarian cancer risk was found for individual GSTM1, GSTA1 and GSTP1 genotypes (p>0.05). However, the carriers of GSTT1-active genotype were at 2-fold higher risk of ovarian cancer development (95%C1: 1.00-4.01, p=0.049), which was even more elevated in the subgroup of patients with positive family history of cancer. Moreover, the frequency of all three GST genotypes that might be associated to ovarian cancer risk (GSTT1-active, GSTA1-active and GSTPl-referent) was significantly higher in patients than in the control group (p=0.042). Even more, patients who were carriers of combination of these three genotypes represented over 64% of the total number of patients within any of the International Federation of Gynecology and Obstetrics (FIGO) stages of ovarian cancer. Conclusions: This study provides supportive evidence that GSTs might affect both susceptibility and progression of ovarian cancer.

Purpose: Since several studies have proposed that epithelial ovarian cancer should not be considered as a single disease entity and that it results from an accumulation of genetic changes, we aimed to assess the polymorphic expression of major cytosolic glutathione S-transferases (GSTM1, T1, A1 and P1) with respect to ovarian cancer susceptibility and aggressiveness. Methods: This case-control study was conducted on 93 newly diagnosed epithelial ovarian cancer patients and 178 healthy matched controls. The multiplex polymerase chain reaction (PCR) was used to detect homozygous deletions ofGSTMl and GSTT1 genes. Analysis of the single nucleotide polymorphism (SNP) GSTA1 C69T was performed using PCR-restrictionfragment length polymorphism (RFLP), while for SNP GSTP1 Ile105Val real-time PCR was used. Results: No significant association to ovarian cancer risk was found for individual GSTM1, GSTA1 and GSTP1 genotypes (p>0.05). However, the carriers of GSTT1-active genotype were at 2-fold higher risk of ovarian cancer development (95%C1: 1.00-4.01, p=0.049), which was even more elevated in the subgroup of patients with positive family history of cancer. Moreover, the frequency of all three GST genotypes that might be associated to ovarian cancer risk (GSTT1-active, GSTA1-active and GSTPl-referent) was significantly higher in patients than in the control group (p=0.042). Even more, patients who were carriers of combination of these three genotypes represented over 64% of the total number of patients within any of the International Federation of Gynecology and Obstetrics (FIGO) stages of ovarian cancer. Conclusions: This study provides supportive evidence that GSTs might affect both susceptibility and progression of ovarian cancer.

Purpose: Matrix metalloproteinases (MMPs) are a family of endopeptidases that may play an important role in the development of salivary gland cancer (SGC). MMP-2 and MMP-9, members of the gelatinase protein family, are capable of degrading type IV collagen of basement membranes, and their overexpression is often associated with tumor aggressiveness and poor prognosis. The aim of this study was to establish the role of single nucleotide polymorphisms (SNPs) in MMP-2 and MMP-9 genes as putative susceptibility factors for the development of SGC. Methods: The MMP-2-1306 C>T, MMP-2-1575 G>A and MMP-9-1562 C>T polymorphisms were analyzed in 93 SGC cases and 100 controls using PCR-RFLP. Results: The T allele for the MMP-2-1306 C>T polymorphism exhibited its effect in heterozygous carriers, increasing the risk for SGC (odds ratio/OR 1.98, 95% CI 1.07-3.65, p=0.03). According to the dominant model, CT+TT genotypes had a 2-fold increased risk of developing SGCs (p=0.02). When the dominant model was applied for the MMP2-1575 G>A, individuals with GA+AA genotypes exhibited a 1.77-fold increase in cancer risk, but with borderline significance (p=0.049). Heterozygous carriers of the variant T allele for the MMP-9-1562 C>T polymorphism had roughly a 2-fold increase in susceptibility for SGC compared to wild type homozygotes (CC) (p=0.02). Conclusion: Our findings suggest MMP-2-1306 C>T and MMP-9-1562 C>T polymorphisms genotypes seem to influence the development of SGCs, whereas MMP-2-1575 G>A seems to be of a minor importance.

Purpose: Matrix metalloproteinases (MMPs) are a family of endopeptidases that may play an important role in the development of salivary gland cancer (SGC). MMP-2 and MMP-9, members of the gelatinase protein family, are capable of degrading type IV collagen of basement membranes, and their overexpression is often associated with tumor aggressiveness and poor prognosis. The aim of this study was to establish the role of single nucleotide polymorphisms (SNPs) in MMP-2 and MMP-9 genes as putative susceptibility factors for the development of SGC. Methods: The MMP-2-1306 C>T, MMP-2-1575 G>A and MMP-9-1562 C>T polymorphisms were analyzed in 93 SGC cases and 100 controls using PCR-RFLP. Results: The T allele for the MMP-2-1306 C>T polymorphism exhibited its effect in heterozygous carriers, increasing the risk for SGC (odds ratio/OR 1.98, 95% CI 1.07-3.65, p=0.03). According to the dominant model, CT+TT genotypes had a 2-fold increased risk of developing SGCs (p=0.02). When the dominant model was applied for the MMP2-1575 G>A, individuals with GA+AA genotypes exhibited a 1.77-fold increase in cancer risk, but with borderline significance (p=0.049). Heterozygous carriers of the variant T allele for the MMP-9-1562 C>T polymorphism had roughly a 2-fold increase in susceptibility for SGC compared to wild type homozygotes (CC) (p=0.02). Conclusion: Our findings suggest MMP-2-1306 C>T and MMP-9-1562 C>T polymorphisms genotypes seem to influence the development of SGCs, whereas MMP-2-1575 G>A seems to be of a minor importance.

The study provides evidence on the individual and family factors as potential predictors (odds ratio—OR and 95% CI) of cyber-violence among school-aged children (11–17 years old) from 64 schools participating in the 2017 Serbian Study on health behavior in school-age children (HBSC). The standardized international HBSC research protocol was used. The study population was the nationally representative sample of 3267 students of V and VII grades of primary and I grade of secondary schools in Serbia. Potential predictors for the probability of occurrence vs. non-occurrence of cyberbullying exposure at least once and multiple times were identified among 24 explanatory variables, including the individual characteristics and family context. The cyberbullying exposure was more prevalent among girls than among boys of school-age, i.e., over one in seven girls and one in ten boys were exposed to cyberbullying. Over one in seven students at age 13 years and almost every seventh student at grade I of the gymnasium were exposed to cyberbullying. There were more students exposed to at least one cyberbullying than to multiple cyberbullying. Potential predictors of exposure to cyberbullying are gender, opinion of the family’s affluence status, fathers’ employment, communication with father, and family support. The study compensates for the evidence of cyberbullying in Serbia, which could help raise awareness, inform national and international stakeholders in the region and enable their efforts and strengthen cooperation in ending cyberbullying. This study’s findings could inform the development of an intervention program aimed at families and various professionals involved in protecting and improving school-age children’s health and well-being. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

The study provides evidence on the individual and family factors as potential predictors (odds ratio—OR and 95% CI) of cyber-violence among school-aged children (11–17 years old) from 64 schools participating in the 2017 Serbian Study on health behavior in school-age children (HBSC). The standardized international HBSC research protocol was used. The study population was the nationally representative sample of 3267 students of V and VII grades of primary and I grade of secondary schools in Serbia. Potential predictors for the probability of occurrence vs. non-occurrence of cyberbullying exposure at least once and multiple times were identified among 24 explanatory variables, including the individual characteristics and family context. The cyberbullying exposure was more prevalent among girls than among boys of school-age, i.e., over one in seven girls and one in ten boys were exposed to cyberbullying. Over one in seven students at age 13 years and almost every seventh student at grade I of the gymnasium were exposed to cyberbullying. There were more students exposed to at least one cyberbullying than to multiple cyberbullying. Potential predictors of exposure to cyberbullying are gender, opinion of the family’s affluence status, fathers’ employment, communication with father, and family support. The study compensates for the evidence of cyberbullying in Serbia, which could help raise awareness, inform national and international stakeholders in the region and enable their efforts and strengthen cooperation in ending cyberbullying. This study’s findings could inform the development of an intervention program aimed at families and various professionals involved in protecting and improving school-age children’s health and well-being. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Nodular fasciitis (NF), also called proliferative or infiltrative fasciitis is considered to be a benign reactive process of the soft tissues (subcutaneous tissue, muscle or fascia) rather than a true neoplasm. Although the extremities are the most common sites, it may arise anywhere in the body. Local excision presents the main treatment. This study presents a case of NF with a unique clinical presentation, an acute subcutaneous temporal hemorrhagic growth. Because temporal NF is more often dermally situated than NF involving other anatomic sites, it may present with superficial ulceration or bleeding and appear clinically unusual. Nodular fasciitis should be considered in the differential diagnosis of other benign or malignant tissue masses involving the temporal area. © 2011 European Association for Cranio-Maxillo-Facial Surgery.

Nodular fasciitis (NF), also called proliferative or infiltrative fasciitis is considered to be a benign reactive process of the soft tissues (subcutaneous tissue, muscle or fascia) rather than a true neoplasm. Although the extremities are the most common sites, it may arise anywhere in the body. Local excision presents the main treatment. This study presents a case of NF with a unique clinical presentation, an acute subcutaneous temporal hemorrhagic growth. Because temporal NF is more often dermally situated than NF involving other anatomic sites, it may present with superficial ulceration or bleeding and appear clinically unusual. Nodular fasciitis should be considered in the differential diagnosis of other benign or malignant tissue masses involving the temporal area. © 2011 European Association for Cranio-Maxillo-Facial Surgery.

Reinke's space is a highly specific structure critical for the function of the vocal fold, involved in a majority of pathological changes of the larynx. The aim of the study was to contribute to the understanding of edematous areas of vocal fold mucosa in patients with Reinke's edema using contact telescopy (ConTel). The edematous tissue which could be identified grossly by microlaryngoscopy under general anesthesia was stained topically with methylene blue and then examined using ConTel. The observed changes on contact images were further correlated with corresponding histological sections of biopsied edematous tissue. In patients with Reinke's edema examined using ConTel, we observed longitudinal arranged mucosal "blue lines" (BL). In the histological sections of edematous tissue showing the BL on ConTel, we almost regularly observed well-developed hollow spaces in subepithelia. Also, they were regularly oriented from the middle of the membranous portion of the vocal fold toward the arcuate line distributed almost parallel to the free edge of the vocal fold. BL were a specific feature of Reinke's edema as they were not found in control groups with normal vocal fold mucosa and with vocal fold polyps. For the first time we are describing a new superficial morphological feature of Reinke's edema. The BL could present epithelial ConTel markers of the area between two enlarged neighboring subepithelial crevices of Reinke's space, as indicated on histological sections. Further characterization of BL is required for the understanding of their role in Reinke's edema. © 2009 The Voice Foundation.

Reinke's space is a highly specific structure critical for the function of the vocal fold, involved in a majority of pathological changes of the larynx. The aim of the study was to contribute to the understanding of edematous areas of vocal fold mucosa in patients with Reinke's edema using contact telescopy (ConTel). The edematous tissue which could be identified grossly by microlaryngoscopy under general anesthesia was stained topically with methylene blue and then examined using ConTel. The observed changes on contact images were further correlated with corresponding histological sections of biopsied edematous tissue. In patients with Reinke's edema examined using ConTel, we observed longitudinal arranged mucosal "blue lines" (BL). In the histological sections of edematous tissue showing the BL on ConTel, we almost regularly observed well-developed hollow spaces in subepithelia. Also, they were regularly oriented from the middle of the membranous portion of the vocal fold toward the arcuate line distributed almost parallel to the free edge of the vocal fold. BL were a specific feature of Reinke's edema as they were not found in control groups with normal vocal fold mucosa and with vocal fold polyps. For the first time we are describing a new superficial morphological feature of Reinke's edema. The BL could present epithelial ConTel markers of the area between two enlarged neighboring subepithelial crevices of Reinke's space, as indicated on histological sections. Further characterization of BL is required for the understanding of their role in Reinke's edema. © 2009 The Voice Foundation.