The effect of the alternative solutions which are produced to be used in place of formalin and xylene on tissue processing

Amaç: Formalin ve ksilolün önemli toksik, allerjen ve karsinojen etkileri vardır. Günümüzde, bu kimyasalların yerine geçebilecek daha az toksik maddeler piyasaya sürülmüştür. Bu maddelerin, histokimyasal ve immünohistokimyasal boyamalara; in situ hibridizasyon, polimeraz zincir reaksiyonu gibi incelemelere olumsuz etki etmemesi beklenir. Çalışmamızda, Türkiye'de ticari olarak satılan alternatif kimyasalların etkilerinin araştırılması amaçlanmıştır.  Gereç ve Yöntem: 5 farklı fiksatif (Formalin, Glyo-Fixx, FineFIX, Cell block ve Greenfix) ve 4 farklı şeffaflandırıcı (Ksilen, Sub-X, Bio Clear, Shandon Xylene Substitute) kullanılarak 13 farklı doku takibi kurgulanıp, 13 farklı dokuya uygulandı. Hematoksilen-Eozin boyalı kesitler histomorfolojik düzeyde niteliksel olarak karşılaştırıldı. Hazırlanan doku mikrodizini kesitlerine histokimyasal ve immünohistokimyasal boyamalar uygulandı. Histokimyasal boyamalar niteliksel, immünohistokimyasal boyamalar ise bilgisayar programında (3D Histech NuclearQuant, MembraneQuant) niceliksel olarak karşılaştırıldı. Kromojenik in situ hibridizasyon uygulandı ve sinyal kalitesi değerlendirildi. DNA ve RNA ekstraksiyonu uygulanarak ve DNA niteliği polimeraz zincir reaksiyonu ile değerlendirilerek doku takipleri moleküler düzeyde kıyaslandı. Bulgular: Tüm doku takiplerinde kesit kalitesinin iyi olduğu gözlendi. Histokimyasal boyama sonuçları, alternatif solüsyonların tümünün bu uygulamalar açısından uyumlu olduğunu gösterdi. İmmünohistokimyasal boyamalarda etken maddesi glyoxal olan fiksatiflerle olumsuz sonuçlar elde edildi. Kromojenik in situ hibridizasyon ile tüm doku örneklerinde sinyalin okunaklığı, zeminin kontrastı benzer ve iyi özellikte bulundu. Ayrıca formalin içermeyen takiplerin moleküler korunma açısından, formalin içerenlere göre üstün olduğu görüldü. Sonuç: Tüm parametreler birlikte değerlendirildiğinde, Glyo-Fixx, Cell block ve FineFIX kullanılan doku takiplerinin, biraz daha üstün oldukları dikkati çekmektedir. Daha az toksik, daha az irritan olmaları, iyi morfolojik sonuç vermeleri, histokimyasal boyamalar ile uyumlu olmaları ve moleküler koruma açısından formaline üstün olmaları nedeni ile alternatif fiksatifler, rutin patoloji ve araştırmalarda formalinin yerini almaya adaydır. Alternatif fiksatif arayışının geri planda bıraktığı alternatif şeffaflandırıcıların da ksilenin yerini almaya güçlü adaylar olduğu görülmüştür. Aim: Formaldehyde and xylene are main solutions of tissue processing. But these chemical substances have important toxic, allergen and carcinogenic effects on human body. Nowadays, nontoxic or lesser toxic new commercial solutions have been produced to be used in place of formaldehyde and xylene. It is expected to be compatible with histochemical and immunohistochemical stainings, also with in situ hibridisation and polymerase chain reaction from these new alternative solutions. In this study, we aimed to assess the impact of new alternative solutions on methods mentioned above.  Methods: Thirteen different tissue processings were designed and performed on 13 different tissues by using five different fixatives (Formaldehyde, Glyo-Fixx, FineFIX, Cell block ve Greenfix) and four different clearing agents (Xylene, Sub-X, Bio Clear, Shandon Xylene Substitute). Haematoxylin&Eosine stained sections were compaired by using qualitative histomorfological criterions. Histochemical and immunohistochemical stains were performed by using tissue microarrays. Histochemical staining results were compaired by using qualitative criterions while immunohistochemical staining results were compaired with quantitative datas which were had by using a computer programme (3D Histech NuclearQuant, MembraneQuant). Tissue sections were tested for the availability of chromogenic in situ hibridisation, by using a Her-2/neu protocol. Also different tissue processings were compaired at molecular level by performing DNA and RNA extraction, and DNA quality by polymerase chain reaction. Results: The quality of sections was well for all tissue processings. Histochemical staining results showed that all alternative solutions which were used are suitable for this application. Immunohistochemical staining results showed that alternative solutions which contain glyoxal as active agent need optimization fort his application. The clearance of signals with chromogenic in situ hibridisation were nearly same and well for all tissue samples. Furthermore, tissue processes which do not contain formaldehyde were found to be superior compaired with ones which contain formalin as fixative solution on molecular preservation. Conclusion: We have demonstrated that formaldehyde-free fixatives have potential in routine pathology and research to replace formaldehyde in histomorphology and protein preservation. When all parameters were evaluated together it was found that the tissue processes which contain Glyo-Fixx, Cell block and FineFIX as fixative solution were superior compared the others. We have also demonstrated that alternative clearing agents which stayed behind the scenes because of the search of ideal fixative have the potantial in routine pathology to replace xylene

CD10 Expression in Epithelial and Stromal Cells of Non-small Cell Lung Carcinoma (NSCLC): A Clinic and Pathologic Correlation

CD10 is a zinc dependent metallopeptidase, and its expression in stromal and/or epithelial cells of many carcinomas has been suggested to have prognostic value. This study investigates CD10 expression in epithelial and stromal cells of non small cell lung carcinoma (NSCLC), and evaluates its prognostic value for this tumor and its histologic subtypes. Sixty-six cases of NSCLC [35 cases of nonsquamous cell carcinoma (NSCC) and 31 cases of squamous cell carcinoma (SCC)] were analyzed immunohistochemically for CD10 antibody. Fisher's exact test and univariate survival analyses were performed. Comparison of clinicopathologic characteristics for NSCLC showed that only stromal CD10 expression had worse prognostic impact, associated with the presence of recurrence (p=0.001), death (p=0.006) and disease positivity (p=0.001). For SCC, CD10 was found to be expressed mainly in the stromal cells, and was associated with a decreased survival (p=0.000) and disease free survival (p=0.000). CD10 expression was restricted to the epithelial cells in NSCC and associated with an increased disease free survival (p=0.036). Stromal CD10 expression apppears to be a worse prognostic factor in NSCLCs. CD10 which is expressed in different cell components of SCC and NSCC appears to have opposing effects on the behaviour of these histologic types

Cystic fibroadenoma of the breast: A case report Memenin kistik fibroadenomu: Olgu sunumu

Fibroadenoma is the most common breast tumor in adolescent and young women. Fibroadenomas that consist of sclerosing adenosis, papillary apocrine metaplasia, epithelial calcifications, and/or cysts greater than 3 mm are considered as complex fibroadenoma. The relative risk of developing breast cancer in patients with complex fibroadenoma is increased, compared to women with noncomplex fibroadenoma. Extensive cystic degeneration in a fibroadenoma, so called "cystic fibroadenoma" is very rare. Herein, we present a case of such a lesion in a 43-year-old female who has been on follow-up for fibrocystic changes of the breast, and discuss both radiological and histopathologic diferential diagnosis of this lesion with other cystic lesions of the breast, including cystic papilloma. The patient is free of disease afer 17 months of clinical follow-up

Recurrent Pseudo-TORCH Appearances of the Brain Presenting as "Dandy-Walker" Malformation

Dandy-Walker malformation (DWM) is a developmental abnormality characterized by cystic dilatation of the fourth ventricle, complete or partial agenesis of the cerebellar vermis, and enlarged posterior fossa with upward displacement of the lateral sinuses, tentorium, and torcula. Calcification of brain tissue is a feature of congenital infection, particularly those involving the TORCH (Toxoplasma gondii, rubella virus, Cytomegalovirus, and herpesvirus) group. An autosomal-recessive congenital infection-like syndrome with intracranial and extracranial calcifications has been categorized as pseudo-TORCH syndrome. We describe two male siblings diagnosed as DWM by ultrasound and by in utero and postmortem magnetic resonance imaging (MRI) but in whom the neuropathology revealed features of intracranial necrosis and calcification in the absence of extracranial calcific deposition. The fetal anomaly was identified by routine prenatal ultrasound scan at 16 weeks. In both cases the postmortem MRI showed bilateral ventriculomegaly with distortion of the overlying cortices, enlarged posterior fossa with a cyst related to small cerebellar hemispheres, and an incomplete cerebellar vermis. The diagnosis of DWM was offered. The histology revealed hypoplastic cerebral hemispheres with poorly cellular developing cortex. The white matter and periventricular matrix were disrupted by areas of necrosis and calcification not associated with any inflammatory infiltration, organisms, inclusions, or giant cells. To our knowledge, these two male siblings are the 1st cases that show pseudo-TORCH syndrome with distinctive intracranial calcification presenting as DWM. An autosomal-recessive or X-linked inheritance needs to be considered. Our study confirms the relevance of the multidisciplinary teamwork involved in the diagnosis of these complex cases

PİLONİDAL SİNÜS AMELİYATI SONRASI FARKLI YATIŞ POZİSYONUNUN DRENAJ MİKTARI ÜZERİNE ETKİSİ

Amaç: Bu çalışma pilonidal sinüs ameliyatı sonrası farklı yatış pozisyonunun drenaj miktarı üzerine etkisini belirlemek amacıyla girişimsel olarak yürütüldü. Yöntem: Çalışmanın evrenini Nisan 2014 – Mayıs 2015 tarihleri arasında Kars Harakani Devlet Hastanesi Genel Cerrahi Servisinde pilonidal sinüs ameliyatı planlanan hastalar oluşturdu. Örneklemi ise; belirtilen evrenden olasılıksız rastlantısal örnekleme yöntemi ile araştırmaya alınma kriterlerini taşıyan 60 hasta (30 yüzüstü, 30 sol lateral/yarı sırtüstü yatış grubu) oluşturdu. Veri toplamada hastalar için “Tanıtıcı Özellikler Formu” ve “Pilonidal Sinüs Eksizyonu Postoperatif Drenaj Miktarı Takip Formu” kullanıldı. Verilerin analizinde tanımlayıcı istatistikler, ki-kare (x2 ), Mauchly’s W ve t testi yapıldı. Bulgular: Çalışmada grupların tanıtıcı özellikler yönünden homojen olduğu tespit edildi. Kontrol grubunda ameliyat sonrası yüzüstü pozisyonunda yatırılan hastalarda ortalama 24. saat sonunda 25.16±15.32, 48. saat sonunda 4.80±3.82, 72. saat sonunda 1.03±2.04, toplam 30.96±19.35 olarak bulundu. Deney grubunda ameliyat sonrası sol lateral/yarı sırtüstü pozisyonunda yatırılan hastalarda ortalama 24. saat sonunda 17.76±8.83, 48. saat sonunda 3.26±2.72, 72. saat sonunda 1.10±2.00, toplam 22.13±11.56 olarak saptandı. Sonuç: Çalışma sonucuna göre ameliyat sonrası yüzüstü pozisyonunda yatırılan pilonidal sinüs hastalarının drenaj miktarı sol lateral/yarı sırtüstü pozisyonunda yatan pilonidal sinüs hastalarından daha fazla olduğu görüldü.Aim: This study was conducted interventionally to determine the effect of different lying position on drainage amount after pilonidal sinus surgery. Methods: the study population consisted of patients with pilonidal sinus who were hospitalized in Kars Harakani State Hospital General Surgery Department between April 2014 and May 2015. the sample is; 60 patients (30 prone, 30 left lateral/semi supine hospitalization group) who met the inclusion criteria by random sampling method. “Introductory Features Form” and “Pilonidal Sinus Excision Postoperative Drainage Amount Tracking Form” were used for data collection. Descriptive statistics, chi-square (x2 ), Mauchlys W and t tests were used to analyze the data. Results: in the study, it was determined that the groups were homogeneous. in the control group, 25.16 ± 15.32 at the end of the 24th hour and 4.80 ± 3.82 at the 48th hour and 1.03 ± 2.04 at the 72th hour and 30.96 ± 19.35 at the end of the operation. in the experimental group, 17.76 ± 8.83 at the end of 24 hours, 3.26 ± 2.72 at the 48th hour, 1.10 ± 2.00 at the end of the 72th hour and a total of 22.13 ± 11.56 at the end of the operation. Conclusion: According to the results of the study, the amount of drainage of the pilonidal sinus patients in the postoperative position prone was higher than the pilonidal sinus in the left lateral/semi supine