PRAME and Historical Immunohistochemical Antibodies Ki-67, P16, and HMB-45 in Ambiguous Melanocytic Tumors.

ABSTRACT: Ambiguous melanocytic lesions/tumors (AMLs) can be simply described as melanocytic neoplasms that cannot be differentiated as either a melanoma or a nevus. Preferentially expressed antigen in melanoma (PRAME) is a novel antibody that can help differentiate between nevi and melanomas. However, its usefulness remains controversial in AMLs. The aim of this study was to demonstrate the importance of PRAME and diagnostic auxiliary antibodies (Ki-67, p16, HMB-45) in the diagnosis of melanocytic lesions, especially in AMLs. This study included 52 ambiguous melanocytic lesions, 40 nevi, and 40 melanomas. All immunohistochemical studies were performed automatically using the Universal Alkaline Phosphatase Red Detection Kit. Different analytic approaches were used for each antibody based on the literature. Statistically, the multinomial forward stepwise elimination logistic regression analysis was used to create a statistical model to predict the diagnosis of melanocytic lesions based on clinical, morphological, and immunohistochemical data. PRAME positivity was very strong and diffuse in the melanoma group and statistically significantly higher than that of the AML and nevus groups. There was no statistically significant difference between the nevus and AML groups. The Ki-67 proliferation index and HMB-45 staining pattern provided valuable indications for distinguishing between these 3 groups. The P16 antibody was limited in supporting the differential diagnosis. Our statistical model showed that a high mitosis count, central pagetoid spread, and PRAME positivity increased the probability of melanoma against an AML diagnosis. This study showed the advantages of evaluating the PRAME antibody together with morphological features and other immunohistochemical markers (Ki-67 and HMB-45) in the differential diagnosis of melanocytic lesions.

The effect of the platelet-rich plasma on osteogenic potential of the periosteum in an animal bone defect model.

OBJECTIVES: This study aims to investigate whether plasma-rich plasma (PRP) enhances the osteogenic potential of periosteal grafts used to repair bone defects and maintains both histologically and biomechanically more durable bone tissue. MATERIALS AND METHODS: A standard bone defect was formed to the left femurs of 54 Sprague-Dawley rats and three groups were formed. In the first group (n=18), no periosteal repair was done for bone defect. In the second group (n=18), periosteal graft tissue was sutured to cover the defect entirely. In the third group (n=18), before periosteal repair, a 1 mL of PRP fibrin was applied into the bone defect. All femoral specimens were compared histologically at four and six weeks and biomechanically by three-point bending test at six weeks after treatment. RESULTS: In the PRP applied group, healing of the bone defect at four weeks was significantly better than the other groups in terms of histological new bone formation (p<0.05). At six weeks, new bone formation in both of the periosteum preserved groups was superior to the first group (p<0.05, for both). There was no statistically significant difference between the second and third groups at the end of the sixth week in the biomechanical analysis, although both groups were significantly stronger than the first group (p<0.05). CONCLUSION: Stimulation of the periosteum with PRP application causes early osteogenic differentiation of precursor cells. Although, at biomechanical basis, PRP application does not create any significant difference, in the recovery of the bone defects at very early period, application of PRP may play a role to accelerate fracture healing and to decrease nonunions.

Kindlin-3 and RASSF6 are probable biomarkers for predicting metastasis in cutaneous melanoma.

It is well known that metastasis is the most crucial factor in determining the fate of the patient. The prognosis of melanoma is very poor at the stage of metastasis. Recently, several genes and proteins, including kindlin3, dioxin receptor (AhR), RASSF6, and claudin-11, which were shown as possible prognostic biomarkers for human tumours, were described. In this study, we focused on these proteins in melanoma within a clinical setting. Forty-three primary melanomas (PMs), 17 metastatic melanomas (MMs), 15 melanocytic nevi (MN), and two melanoma cell lines were included in this retrospective study. All proteins were investigated using immunohistochemistry, and analysis was performed using a semi-quantitative immunoreactive score (IRS). The nevus group showed lower RASSF6 and AhR IRS levels than PMs. RASSF6 and kindlin-3 levels in the PMs with metastasis (MwM) and also in PMs showing lymphovascular invasion were significantly lower. The logistic regression model also proved that kindlin-3 expression was a significant independent predictor of metastasis. The current study supports the role of kindlin-3 and RASSF6 as prognostic biomarkers in melanoma. Besides the prognostic roles of these proteins, they are probably potential candidates for target-oriented therapies for melanoma metastasis blocking.

Primary low grade myxoid liposarcoma of the ovary: A case report and review of literature.

A 20-year-old female patient presented with the complaint of pelvic pain. Radiological studies showed a cystic mass in the left ovary. Histological examination revealed a myxoid tumor with a chicken wire-like, thin vascularization. The diagnosis of the lesion, morphologically resembling myxoid liposarcoma was supported by demonstration of rearrangement of the DDIT3 gene by fluorescence in situ hybridization.

Are there EWSR1 rearranged cutaneous hidradenomas and mucoepidermoid carcinomas of salivary glands? A FISH study and review of the literature.

MAML2 rearrangements have been previously described for hidradenomas and muco-epidermoid carcinomas (MEC). However, one report showed EWSR1 rearrangement in both tumours. In this study, EWSR1 and MAML2 translocation were investigated in hidradenomas and MECs. Specimens from thirteen MECs of the salivary glands and twenty hidradenomas of the skin were evaluated. Fluorescence in situ hybridisation (FISH) studies with EWSR1 and MAML2 break-apart probes were used. Forty percent of hidradenomas and 84.6% of MECs showed a positive MAML2 break signal. EWSR1 break signal was absent in hidradenomas. Only two MECs showed positive EWSR1 signal and were, thus, reclassified as clear cell carcinoma (CCC). A statistically significant relationship was also observed between clear cells containing hidradenomas and the FISH status. Despite the previous study, EWSR1 translocations could not be established in hidradenomas and MECs. The study further suggests that evaluation of EWSR1 might be obligatory for the correct diagnosis of MAML2-negative MECs to exclude the chance of CCCs. The present study also supports the notion that MAML2 can be used as a marker for hidradenomas and MECs.

Is there any correlation among MKK4 (mitogen-activated protein kinase kinase 4) expression, clinicopathological features, and KRAS/NRAS mutation in colorectal cancer.

BACKGROUND: We aimed to evaluate the correlation between MKK4 expression and clinicopathological features, KRAS/NRAS mutation in colorectal cancer. METHODS: MKK4 expression was assessed by immunoreactivity score (IRS). Staining intensity(SI) and percentage of positively stained cells (PP) were used for IRS (IRS = SI×PP). Cutoffs were explored with ROC analysis. Patients were grouped as WIR ('weak immunoreactive'; IRS:0-2) and SIR ('strong immunoreactive'; IRS: >3). RESULTS: We enrolled 95 patients. 63.2% had metastasis. Median follow-up was 31.4 months. KRAS/NRAS mutation rate was 45.2%. Median values for OS, DFS, and PFS were as 31.6, 17.2, and 10.3 months. WIR group had longer OS (p = 0.03). Recurrence rate was 36.8%. Median DFS was longer for recurrent patients in WIR group (p = 0.055). KRAS or NRAS wild-type patients and those with left-sided tumors in WIR group had longer OS (p = 0.029, p = 0.024, p = 0.03). There was no PFS difference (p: 0.15). In correlation analysis, there was a negative correlation between MKK4 expression and KRAS mutation, NRAS mutation, OS, PFS, DFS (r: -0,06; r: -0,02; r: -0,10; r: -0,06; r: -0,34). Only the correlation for MKK4 expression and DFS was significant (p = 0.04). CONCLUSION: MKK4 expression inversely correlates with survival outcomes. Patients with KRAS/NRAS wild-type, left-sided tumors with WIR had longer OS.

Prevention of the harmful effects of free oxygen radicals by using N-acetylcysteine in testicular torsion.

INTRODUCTION: Testicular torsion is a urological emergency both in childhood and in adult life. Many studies on experimental testicular torsion have demonstrated biochemical and pathological ischemia-reperfusion injury and the efficacy of some drugs have been investigated to prevent this damage. N-acetylcysteine (NAC) promotes glutathione synthesis and acts as a glutathione precursor because of the fact that it increases the glutathione-reductase activity by transporting sulfhydryl groups. AIM: In this experimental study, the authors aimed to investigate the effectiveness of NAC in preventing ischemia-reperfusion injury following testicular torsion and detorsion. STUDY DESIGN: For this experimental study, 36 albino Wistar-male rats were used. The rats were randomly divided into 4 groups: sham (n = 8), ischemia-reperfusion (n = 8), ischemia-NAC -reperfusion (n = 10), and ischemia-NAC-reperfusion-NAC (n = 10) groups. Two hours of torsion and 4 h of detorsion were created in the left testis. After 4 h of detorsion, the rats were sacrificed. Each tissue was divided into two sections for biochemical and pathological examinations. RESULTS: There was a statistically significant difference between the study groups in terms of the total-sulfhydryl level, nitric oxide level, and the malondialdehyde values. Histopathological examination revealed that NAC was effective in preventing reperfusion injury in the testis but ineffective in preventing the reduction in the spermatid count. DISCUSSION: The results of this experimental study support that NAC can histopathologically maintain the structure of seminiferous tubules against ischemis reperfusion injury and prevent damage to the germinative cells. However, it was unable to prevent the reduction in spermatid count. There was no significant difference in the prevention of ischemia-reperfusion injury between NAC administration during the first hour of ischemia and NAC administration during reperfusion. Although NAC can prevent tissue damage from ischemia reperfusion injury, it is not effective against the reduction in the spermatid count. CONCLUSION: N-acetylcysteine may be biochemically effective in preventing ischemia-reperfusion injury after testicular torsion and detorsion. NAC is a readily available and easy to use agent that can be used during testicular ischemia.

Hypoxic Gene Signature of Primary and Metastatic Melanoma Cell Lines: Focusing on HIF-1ß and NDRG-1

Background: Hypoxia is an important microenvironmental factor significantly affecting tumor proliferation and progression. The importance of hypoxia is, however, not well known in oncogenesis of malignant melanoma. Aims: To evaluate the difference of hypoxic gene expression signatures in primary melanoma cell lines and metastatic melanoma cell lines and to find the expression changes of hypoxia-related genes in primary melanoma cell lines at experimental hypoxic conditions. Study Design: Cell study. Methods: The mRNA expression levels of hypoxia-related genes in primary melanoma cell lines and metastatic melanoma cell lines and at experimental hypoxic conditions in primary melanoma cell lines were evaluated by using real-time polymerase chain reaction. Depending on the experimental data, we focused on two genes/proteins, the hypoxia-inducible factor-1 beta and the N-myc downstream regulated gene-1. The expression levels of the two proteins were investigated by immunohistochemistry methods in 16 primary and metastatic melanomas, 10 intradermal nevi, and a commercial tissue array comprised of 208 cores including 192 primary and metastatic malignant melanomas. Results: The real-time polymerase chain reaction study showed that hypoxic gene expression signature was different between metastatic melanoma cell lines and primary melanoma cell lines. Hypoxic experimental conditions significantly affected the hypoxic gene expression signature. In immunohistochemical study, N-myc downstream regulated gene-1 expression was found to be lower in primary cutaneous melanoma compared to in intradermal nevi (p=0.001). In contrast, the cytoplasmic expression of hypoxia-inducible factor-1 beta was higher in primary cutaneous melanoma than in intradermal nevi (p=0.001). We also detected medium/strong significant correlations between the two proteins studied in the study groups. Conclusion: Hypoxic response consists of closely related proteins in more complex pathways. These findings will shed light on hypoxic processes in melanoma and unlock a Pandora's box for development of new therapeutic strategies.

Juvenile-Onset Early-Stage Mycosis Fungoides-Associated Follicular Mucinosis: A Case Report.

We report a juvenile case of mycosis fungoides with prominent follicular mucinosis (FM). The patient was a 9-year old boy who presented with a 2-month history of enlarging alopecic patch with fine scales on the scalp. Dermatologic examination revealed orange-tan slightly palpable plaques with follicular prominence on his trunk. The patient and his family were not aware of these asymptomatic truncal plaques. Histopathologic examination of both-scalp and trunk-lesions revealed folliculotropic lymphocytic infiltration with mucin. Immunohistochemical study showed that lymphocytic infiltration was CD4 dominant. Flow cytometry analyses of peripheral blood were normal. Any abnormal populations and Sézary cells were not observed on blood smear. Polymerase chain reaction testing showed monoclonality for the T-cell receptor4-[Latin Small Letter Rams Horn] gene. Our patient had the clinical and histopathological diagnosis of follicular mycosis fungoides-associated follicular mucinosis.

Transducing-Like Enhancer of Split 1: A Potential Immunohistochemical Marker for Glomus Tumor.

Glomus tumors (GTs) are rare, perivascular soft tissue tumors. Although GTs are usually found in the subcutaneous tissue, they may be detected in extracutaneous sites and mucosal areas. Transducing-like enhancer of split 1 (TLE1) is a highly useful immunohistochemical marker, which basically helps in differential diagnosis of synovial sarcoma. Based on a coincidental detection of TLE1 in one GT case, we studied 26 additional GT cases to establish the importance and distribution of TLE1 in GTs. Of 24 subcutaneous GTs, 22 (91.6%) were positive for TLE1 antibody and the remaining 2 mucosal GTs were negative. Of the 22 positive cases, 10 showed strong nuclear positivity. There was no difference between the subtypes of the GTs. Although TLE1 expression is significantly correlated to SS18 (SYT) rearrangements in synovial sarcomas, the fluorescence in situ hybridization analyses of the GTs showed no evidence of translocation involving this locus. TLE1 is a potential immunohistochemical marker for GTs, but further studies are required to confirm this finding.

Two Different Cell Populations Is an Important Clue for Diagnosis of Primary Cutaneous Adenoid Cystic Carcinoma: Immunohistochemical Study.

Primary cutaneous adenoid cystic carcinoma (PCACC) is a very rare malignancy. The differential diagnosis of PCACCs in pathology practice can be difficult and a group of primary and metastatic lesions, including adenoid basal cell carcinoma of the skin, should be considered in the differential diagnosis. Besides histomorphological clues, immunohistochemistry studies are very helpful in the differential diagnosis of PCACC. We report herein a case of PCACC with extensive immunohistochemical studies and review the literature from an immunohistochemistry perspective.

Coexistence of Adult Onset Eruptive Syringoma and Bilateral Renal Cell Carcinoma: A Case Report.

Eruptive syringoma is an unusual variant of syringoma, which usually presents before or during puberty. It typically occurs in large numbers as multiple yellow-brown-colored papules, which may show spontaneous regression. Because some authors have proposed that it could present as a reactive process of eccrine ducts to an inflammatory reaction caused by an unknown trigger, the exact pathomechanism is still unclear. There are also reports in the literature on the association of eruptive syringoma in Down syndrome, diabetes, milium, sarcoidosis, and psychiatric disorders. Some reports in the literature highlighted the association of eruptive syringomas and neoplasms. We describe here a case of adult-onset eruptive syringoma in a 53-year-old man and discuss the possibility of its association with renal cell carcinoma as a paraneoplastic phenomenon.

The effect of low-dose methotrexate on autologous fat graft survival.

BACKGROUND/AIM: The survival of autologous fat graft tissue is dependent on various factors such as vascularization and inflammation. The aim of the present study was to evaluate the possible beneficial effects of low-dose methotrexate (LD-MTX) on fat graft volume and survival. MATERIALS AND METHODS: A total of 13 male Wistar albino rats were divided into two groups, a control group and an LD-MTX group. An autologous fat graft obtained from the inguinal region of each rat was transferred to its back. LD-MTX was administered intraperitoneally in the LD-MTX group once a week for 4 weeks after the surgical procedure. The control group underwent surgery but was not administered MTX. Fat grafts were harvested for analyses. RESULTS: The results showed that 2 months postoperatively the fat graft weights of the control and LD-MTX groups were not significantly different. In addition, the vascularity of the grafts was higher in the LD-MTX group than it was in the control group. The mean lipid peroxidation levels were essentially the same in the two groups, but myeloperoxidation was significantly lower in the LD-MTX group than it was in the other group. CONCLUSION: The results showed that LD-MTX administration may not preserve the quality and volume of transplanted fat tissue in rats.

Metastasis suppressor proteins in cutaneous squamous cell carcinoma.

Cutaneous squamous cell carcinomas (cSCCs) are common human carcinomas. Despite having metastasizing capacities, they usually show less aggressive progression compared to squamous cell carcinoma (SCC) of other organs. Metastasis suppressor proteins (MSPs) are a group of proteins that control and slow-down the metastatic process. In this study, we established the importance of seven well-defined MSPs including NDRG1, NM23-H1, RhoGDI2, E-cadherin, CD82/KAI1, MKK4, and AKAP12 in cSCCs. Protein expression levels of the selected MSPs were detected in 32 cSCCs, 6 in situ SCCs, and two skin cell lines (HaCaT, A-431) by immunohistochemistry. The results were evaluated semi-quantitatively using the HSCORE system. In addition, mRNA expression levels were detected by qRT-PCR in the cell lines. The HSCOREs of NM23-H1 were similar in cSCCs and normal skin tissues, while RGHOGDI2, E-cadherin and AKAP12 were significantly downregulated in cSCCs compared to normal skin. The levels of MKK4, NDRG1 and CD82 were partially conserved in cSCCs. In stage I SCCs, nuclear staining of NM23-H1 (NM23-H1nuc) was significantly lower than in stage II/III SCCs. Only nuclear staining of MKK4 (MKK4nuc) showed significantly higher scores in in situ carcinomas compared to invasive SCCs. In conclusion, similar to other human tumors, we have demonstrated complex differential expression patterns for the MSPs in in-situ and invasive cSCCs. This complex MSP signature warrants further biological and experimental pathway research.

Eruptive Disseminated Porokeratosis in a Patient With Type 2 Diabetes Mellitus.

Porokeratosis includes a group of heterogenous disorders that represents distinct clinical subtypes of the same genetic pattern. Although entire pathogenesis of porokeratosis still remains unknown, certain factors including ultraviolet radiation and immunosuppression are suggested to be some of the factors inducing this disorder. Eruptive disseminated porokeratosis is a recently described form of porokeratosis, which frequently occurs in the presence of immunosuppresion or malignancy. The authors report here a unique case with sudden onset eruptive disseminated porokeratosis associated with diabetes mellitus.

Effectiveness of Dimethylsulfoxide on the Survival and Volume Preservation of Autologous Fat Graft Tissue: A Preliminary Study.

BACKGROUND: The survival of autologous fat graft tissue is dependent on various factors, such as vascularization and inflammation. OBJECTIVE: This study aims to evaluate the possible beneficial effects of dimethylsulfoxide (DMSO) on fat graft volume and survival. METHODS: Eighteen male Wistar albino rats were divided randomly into three groups. An autologous fat graft obtained from the inguinal region of each rat was transferred to its back. DMSO was administered intraperitoneally (IP) in the DMSO-IP group and cutaneously (C) in the DMSO-C group once daily for 15 days after the surgical procedure. The control group underwent surgery but was not administered with DMSO. Two months after surgery, the grafted fatty tissues were harvested for histopathological and biochemical analyses. RESULTS: The results showed that 2 months postoperatively, fat grafts of the DMSO-C and DMSO-IP groups weighed significantly more than the grafts of the control group. Moreover, the vascularity of the grafts was higher in the DMSO-C group than in the control group, and no significant difference was found between the two DMSO groups. The mean lipid peroxidation levels were the same in the three groups, but myeloperoxidation was significantly lower in the DMSO-C group than in the other two groups. CONCLUSIONS: The study results showed that cutaneous rather than intraperitoneal DMSO administration could preserve the quality and volume of transplanted fat tissue in rats by enhancing vascularity and decreasing inflammation.

Transgelin gene is frequently downregulated by promoter DNA hypermethylation in breast cancer.

BACKGROUND: CpG hypermethylation in gene promoters is a frequent mechanism of tumor suppressor gene silencing in various types of cancers. It usually occurs at early steps of cancer progression and can be detected easily, giving rise to development of promising biomarkers for both detection and progression of cancer, including breast cancer. 5-aza-2'-deoxycytidine (AZA) is a DNA demethylating and anti-cancer agent resulting in induction of genes suppressed via DNA hypermethylation. RESULTS: Using microarray expression profiling of AZA- or DMSO-treated breast cancer and non-tumorigenic breast (NTB) cells, we identified for the first time TAGLN gene as a target of DNA hypermethylation in breast cancer. TAGLN expression was significantly and frequently downregulated via promoter DNA hypermethylation in breast cancer cells compared to NTB cells, and also in 13/21 (61.9 %) of breast tumors compared to matched normal tissues. Analyses of public microarray methylation data showed that TAGLN was also hypermethylated in 63.02 % of tumors compared to normal tissues; relapse-free survival of patients was worse with higher TAGLN methylation; and methylation levels could discriminate between tumors and healthy tissues with 83.14 % sensitivity and 100 % specificity. Additionally, qRT-PCR and immunohistochemistry experiments showed that TAGLN expression was significantly downregulated in two more independent sets of breast tumors compared to normal tissues and was lower in tumors with poor prognosis. Colony formation was increased in TAGLN silenced NTB cells, while decreased in overexpressing BC cells. CONCLUSIONS: TAGLN gene is frequently downregulated by DNA hypermethylation, and TAGLN promoter methylation profiles could serve as a future diagnostic biomarker, with possible clinical impact regarding the prognosis in breast cancer.