URINE CYTOLOGY AND AXILLARY TESTING FOR INTERPRETATION AND FOLLOW-UP OF URINARY TUMORS.

Microscopic appearance of cells in urine cytological samples is the formal diagnostic approach adjunct to cystoscopy for the detection and follow-up of urinary tumors. However, cystoscopy is a surgical method and cytology may miss low-grade papillary tumors. Several assays and markers have been developed to assist in this. When combined with conventional cytology, uro-oncological diagnostic performance is improved. We review the value of these non-invasive modalities in comparison with urine cytomorphology in the work-up of urothelial malignancies.

Diagnosis and immunocytochemical characterization of an EBV related diffuse B large cell lymphoma of the retroperitoneum from cell block preparations obtained by means of EUS guided FNA.

Diffuse large B cell lymphoma (DLBCL) in the retroperitoneum has been scarcely reported and the diagnostic workup is challenging due to difficulties in access to tissue. A 73-year-old female presented to the gastroenterology clinic, with recent-onset atypical abdominal pain and alternating constipation and diarrhea.

Abdominal primary extra-gastrointestinal stromal tumor (EGIST). A cytologic diagnosis in ascitic fluid.

Dear Editor, We present a case of a primary abdominal Extra-gastrointestinal stromal tumor (E-GIST) in a75 years old female patient. E-GISTs are rare, non-epithelial, mesenchymal tumors arising from the soft tissues of the abdomen - mesentery and retroperitoneum. These tumors are histologically and cytologically similar to the stromal tumors of gastrointestinal tract, composed of round or fusiform cells or a mixture of both in a myxoid background. The recognition of these tumors is important because of their aggressive biological behavior, the metastatic potential and the high rate of recurrence.

Syndecan-2 is a key regulator of transforming growth factor beta 2/Smad2-mediated adhesion in fibrosarcoma cells.

Fibrosarcoma is a rare malignant tumor originating from fibroblasts. Transforming growth factor beta 2 (TGFß2) has been established to regulate processes correlated to fibrosarcoma tumorigenesis. In this study, we investigated the possible participation of syndecan-2 (SDC-2), a cell membrane heparan sulfate (HS) proteoglycan on these TGFß2 functions. Our results demonstrate that the inhibition of SDC-2 expression by short interfering RNA (siSDC2) abolished TGFß2-dependent HT1080 cell adhesion (P ≤ 0.01). In parallel, the downregulation of SDC-2 significantly inhibited TGFß2-induced Smad2 phosphorylation (P ≤ 0.01). The immunoflourescence signal of TGF receptor III as well as its protein expression was decreased in SDC-2-deficient cells. The enhancement of adhesion molecules integrin ß1 (P ≤ 0.01) and focal adhesion kinase expression, induced by TGFß2 treatment (P ≤ 0.001), was markedly inhibited in SDC-2-defficient cells (P ≤ 0.01). Conclusively, the obtained data suggest that SDC-2 modulates TGFß2 transcriptional regulation via Smad signaling to facilitate fibrosarcoma cell adhesion.

History of endometriosis may adversely affect the outcome in menopausal recipients of sibling oocytes.

Due to the known adverse effect of endometriosis on gamete quality, it has always been difficult to demonstrate a direct effect of endometriosis on implantation. In order to eliminate these confounding effects, this prospective comparative study studied a population of menopausal recipients with and without endometriosis sharing sibling oocytes coming from the same donor. The aim was to understand the impact of endometriosis on implantation, pregnancy and live birth rates in menopausal recipients. A total of 240 menopausal recipients of donated sibling oocytes, were divided in two groups. Group I consisted of 120 recipients diagnosed with endometriosis and group II consisted of 120 controls. The implantation and pregnancy rates were significantly lower in the endometriosis group compared with the control group (23.81% versus 31.48%, P=0.019; 45.00% versus 58.33%, P=0.039, respectively). In oocyte donation cycles, a recipient's history of endometriosis might have a negative impact on implantation, pregnancy and live birth rates, even in menopausal women. Infertility in endometriosis may be due to poor oocyte quality or embryos with decreased ability to implant due to impaired fertilization. There are no conclusive data on the impact of endometriosis on implantation. The already-known adverse effect of endometriosis on gamete quality makes it more difficult to demonstrate a direct effect of endometriosis on implantation. In order to eliminate these confounding effects we studied a population of menopausal recipients with and without endometriosis sharing sibling oocytes coming from the same oocyte donor. The oocyte donation model was used in an attempt to understand whether the endometrium, the oocytes or both are affected by endometriosis. The aim of the present study was to understand the impact of endometriosis on implantation, pregnancy and live birth rates in menopausal recipients. A total of 240 menopausal recipients of donated sibling oocytes were divided into two groups. Group I consisted of 120 recipients diagnosed with endometriosis and group II consisted of 120 controls. The pregnancy and implantation rates were significantly lower in the endometriosis group compared to the control group (45.00% versus 58.33%, P=0.039) and (23.81% versus 31.48%, P=0.019) respectively. In oocyte donation cycles, a recipient's history of endometriosis might have a negative impact on implantation, pregnancy and live birth rates, even in menopausal women.

Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles.

In conventional IVF cycles with total fertilization failure, rescue intracytoplasmic sperm injection (ICSI) performed 24h after insemination has yielded poor results. However, when ICSI is used, total fertilization failure is a rare event. The aim of the present study is to investigate the degree of sperm contribution to fertilization failures using the egg-sharing model in oocyte donor cycles. The study included only the oocyte donor cycles of sibling oocytes with total fertilization failure in at least one of the matched recipients. Oocytes from 49 oocyte donor cycles were equally shared among 98 recipients undergoing conventional IVF. Due to total fertilization failure in half of the recipients, rescue ICSI was carried out. Compared with the conventional IVF only group, the rescue ICSI group had a lower pregnancy rate (30.61% versus 71.43%), clinical pregnancy rate (28.57% versus 67.35%) and ongoing pregnancy rate (28.57% versus 63.27%) (all P<0.01). Cryptic sperm defects in apparently normal spermatozoa may be the cause of total fertilization failure, indicating the need for simple routine tests to detect them.

Cytological diagnosis of unifocal Langerhans cell histiocytosis in the temporomandibular joint.

Unifocal Langerhans cell histiocytosis (LCH) presents as a solitary focal bone lesion characterized by clonal proliferation of CD1a (+) and langerin(+) dendritic cells, derived from the bone marrow. In 38-69% of cases BRAF-V600E mutation is revealed. It gains attention to maxillofacial surgeons due to involvement of the orofacial region and early onset of jaws symptoms. If overlooked, massive deconstruction of tissues, occurs. We present a case of LCH of the temporomandibular joint in a 14 aged child where fine needle aspiration favorably contributed in an accurate diagnosis combined with the appropriate clinicoradioliogical findings. This can overrule unwarranted biopsy and monitor the treatment largely where aproach to histopathology services is restricted.

Resveratrol and lycopene ameliorate contrast-induced nephropathy in a rabbit model.

Oxidative stress appears to possess a central role in CIN pathophysiology. Resveratrol (Res) and lycopene (Lyc) are strong natural antioxidants evaluated in a limited number of CIN animal studies in vivo. The aim of the study was to evaluate the potential renoprotective effects of Res/Lyc in a CIN rabbit model. Twenty-four adult male New Zealand white rabbits were equally assigned into four groups: control (saline), CIN (intravenous iopromide; 7.5 g iodine/kg), Res + CIN (per os Res; 5 mg/kg), and Lyc + CIN (per os Lyc; 4 mg/kg). Serum Cr (sCr); symmetric/asymmetric dimethylarginine (SDMA/ADMA); oxidative stress biomarkers: malondialdehyde; total antioxidant capacity; catalase; glutathione) were evaluated in blood samples at three time points: right after (0 h); 24 h; 48 h after iopromide/saline administration. CD20+/CD3+ lymphocytes were determined (48 h). All animals were sacrificed at 48 h and both kidneys collected. Oxidative stress biomarkers were measured in renal tissue. sCr and SDMA/ADMA levels increased significantly in CIN compared to all groups. Oxidative stress secondary to CIN in blood/kidneys was suppressed by Res/Lyc. B and T lymphocytes decreased significantly in CIN compared to all groups. The present study provides emerging evidence that Res/Lyc ameliorate CIN by modulating oxidant/antioxidant balance in blood/renal tissue and by inhibiting vasoconstriction/blood cytotoxicity.

Intraoperative squash Cytology of diffuse glioma not otherwise specified, of the Cerebellum.

OBJECTIVE: Diffuse glioma arises anywhere in the CNS, but most frequent in the cerebral hemispheres. The tumor tends to be seen in children and in younger adults aged 20-30. We report one such case in an older female patient presenting the intraoperative cytology of the tumor. CASE REPORT: A 48-year-old female was diagnosed by MRI with a tumor of cerebellum. Cytologic material was obtained during the resection of the tumor and diagnosed cytologically as glioma. CONCLUSION: This case is presented to focus the ability of the intraoperative cytology in diagnosis of the glioma, using immunocytology and confirmed by histo- immunohistology.

Cerebrospinal fluid and intraoperative squash cytology of childhood ependymoma.

Ependymomas are glial neoplasms of central nervous system originated from the ependymal lining of the brain ventricles and spinal cord central canal, and rarely exfoliated into cerebrospinal fluid (CSF). In this case we report the cytomorphological and immunocytomorphological features of ependymoma in CSF and intraoperative squash preparations, confirmed by histology. Case report. The patient  was a nineteen months old female presented at the University hospital of Heraklion, Crete, in a hemicoma, and was intubated. Computed tomography, scanning and magnetic resonance imaging (MRI), were performed and a mass in the posterior fossa was found. A sample of cerebrospinal fluid (CSF) was sent for cytologic evaluation. A diagnosis of ependymoma was rendered, followed by tumor resection, during which intraoperative squash smears for cytologic interpretation were obtained. Cytological consultation disclosed a grade II ependymoma (WHO grade II), with focally anaplastic features (WHO grade III).

Osteoclast: like giant cell undiferrentiated pancreatic tumor diagnosed by means of EUS guided FNA.

OBJECTIVE: Osteoclast-like giant cell tumours are rare abdominal malignant neoplasms mainly arising in the pancreas. Because of their rarity, clinical and cytopathology reports are very limited, and sonographic features have not been clearly specified ; these tumors are easily misdiagnosed by ultrasound as mucinous cystic tumors (MCTs)  or solid pseudopapillary neoplasms (SPNs). CASE STUDY: We report a case of osteoclast  like giant cell tumor arising in the pancreas of an 80 year old female patient offered by EUS-FNA cytology on direct and cell block slides. A biphasic pattern composed by a malignant mononuclear cell component and a giant cell component were hallmarks to the diagnosis. CONCLUSION: Our case highlights the performance of EUS-FNA in the diagnostic approach of  abdominal tumours and the significance of cell block method in the interpretation of osteoclast-like giant cell pancreatic tumour.

Determination of prenatal exposure to parabens and triclosan and estimation of maternal and fetal burden.

BACKGROUND: Parabens (PBs) and triclosan (TCS) are generally used as antimicrobials mostly in personal care products. Their wide prevalence in daily products raised an acute need for the biomonitoring of these contaminants and the investigation of possible health impacts. MATERIAL AND METHODS: In this study we aimed to quantitatively determine PBs and TCS levels in urine and amniotic fluid samples using a liquid chromatography - mass spectrometry system (LC-MS). Ninety nine (99) pregnant women took part in this research. The samples were collected during the amniocentesis in the early second trimester of their pregnancy. Women of all ages, education, household income and profession were selected. The exposure and the burden of pregnant women and their infants were also evaluated. RESULTS: The most prevalent compound in urine, among the analyzed, was TCS with 74.7 % positive samples while in amniotic fluid methyl paraben (MePB) with 21.2 % positive samples. MePB was detected at higher concentrations in urine (mean: 378.5 ng/mL) followed by TCS (mean: 55.3 ng/mL), ethyl paraben (EtPB) (mean: 23.2 ng/mL) and butyl paraben (BuPB) (mean: 2.3 ng/mL) while benzyl paraben (BePB) was not detected in any urine sample. Concentrations in amniotic fluid samples were much lower. In particular, the mean concentrations were 6.6 ng/mL for MePB, 9.2 ng/mL for EtPB, 0.4 ng/mL for BuPB, 0.6 ng/mL for BePB and 1.8 ng/mL for TCS. The detected levels of all analytes in urine were correlated with those in amniotic fluid but no statistically significant results arose (p >n0.05). Negative associations were observed between amniotic fluid levels of MePB and maternal age (p = 0.05) while both urinary and amniotic levels of TCS were correlated with maternal BMI (p = 0.04). Somatometric characteristics of the infants showed no statistical significant associations with the detected levels of PBs and TCS. CONCLUSION: This study indicated a strong/possible association between exposure of pregnant women to TCS and higher/lower maternal body weight gain during pregnancy. The same trend was observed between amniotic fluid MePB levels and maternal age. However, no statistically significant associations were observed between neonatal somatometric characteristics or health status and PBs and TCS levels.

Apoptosis and cell proliferation correlated with tumor grade in patients with ampullary carcinoma.

OBJECTIVE: To evaluate apoptosis and cell proliferation on cytologic specimens (smears) from endoscopic retrograde cholangiopancreatography in patients with ampulary carcinoma and to correlate that relationship with the grade of the tumors. STUDY DESIGN: Forty patients (23 males and 17 females) aged 45-81 who underwent endoscopic retrograde cholangiopancreatography were diagnosed by cytology as having ampullary adenocarcinoma and the diagnoses were confirmed histologically after an operation. All smears were stained using Papanicolaou and Giemsa stain. Apoptosis was assessed using terminal digoxigenin-labeled dUTP nick-end labeling (TUNEL assay) and cell proliferation using MIB-1 monoclonal antibody by the alkaline phosphatase method. RESULTS: The TUNEL indices were 0.4 +/- 0.07, 0.91 +/- 0.33 and 3.1 +/- 0.9 in well, moderate and poorly differentiated ampullary carcinoma, respectively. The differences in both TUNEL and MIB-1 labeling indices were statistically significant between well, moderately and poorly differentiated ampullary carcinoma, and a positive correlation was found between TUNEL and the MIB-1 indices. CONCLUSION: Apoptosis (cell death) and cell proliferation increase as the grade of the differentiation decreases in ampullary carcinoma, suggesting a rapid turnover of the tumor cells with lower grates of differentiation, and apoptosis may play an important role in the growth of the tumors in patients with ampullary carcinomas.

Carcinogenic, ethanol, acetaldehyde and noncarcinogenic higher alcohols, esters, and methanol compounds found in traditional alcoholic beverages. A risk assessment approach.

Greek fermentation and distillation industries produce traditional spirit beverages, such as tsipouro and tsikoudia, consumed both in bottles and bulk quantities by the general population or tourists. The same spirits are also produced by individuals at home since previous centuries, as a part of the local culture but mainly due to the Greek agricultural sector unique characteristics (small cultivation areas with great number of farmers). In this study, the concentrations of carcinogenic compounds: ethanol and acetaldehyde; and noncarcinogenic: higher alcohols (1-propanol, isobutanol, and isoamyl alcohol), esters (ethyl acetate), and methanol were measured to estimate the potential cancer risk and daily intake of these compounds. The margin of exposure (MOE) of carcinogenic compounds was found to be less than 500 (mean value), well below the toxic threshold of 10,000, above which there is not public concern, as suggested by the European Food Safety Authority. Additionally, through risk assessment of noncarcinogenic compounds, we identified two specific compounds in-bulk spirits (produced by individuals), namely ethyl acetate and isobutanol, with health risk index (HRI) greater than 1 (indicating a possibility to induce side effects by consumption of high amounts). Our results indicate that bottled spirits, which are produced in a controlled environment (alcohol industries), showed higher human safety level in terms of both carcinogenic and noncarcinogenic risk assessment studies, comparing to bulk beverages produced by individuals (with out strict regulations).

Current and Future Trends in Molecular Biomarkers for Diagnostic, Prognostic, and Predictive Purposes in Non-Melanoma Skin Cancer.

Skin cancer represents the most common type of cancer among Caucasians and presents in two main forms: melanoma and non-melanoma skin cancer (NMSC). NMSC is an umbrella term, under which basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and Merkel cell carcinoma (MCC) are found along with the pre-neoplastic lesions, Bowen disease (BD) and actinic keratosis (AK). Due to the mild nature of the majority of NMSC cases, research regarding their biology has attracted much less attention. Nonetheless, NMSC can bear unfavorable characteristics for the patient, such as invasiveness, local recurrence and distant metastases. In addition, late diagnosis is relatively common for a number of cases of NMSC due to the inability to recognize such cases. Recognizing the need for clinically and economically efficient modes of diagnosis, staging, and prognosis, the present review discusses the main etiological and pathological features of NMSC as well as the new and promising molecular biomarkers available including telomere length (TL), telomerase activity (TA), CpG island methylation (CIM), histone methylation and acetylation, microRNAs (miRNAs), and micronuclei frequency (MNf). The evaluation of all these aspects is important for the correct management of NMSC; therefore, the current review aims to assist future studies interested in exploring the diagnostic and prognostic potential of molecular biomarkers for these entities.

The toxic influence and biodegradation of carbon nanofibers in freshwater invertebrates of the families Gammaridae, Ephemerellidae, and Chironomidae.

Carbon nanofibers (CNFs) are widely used in consumer products today. In this study, we assessed the effects of CNFs on the digestive system of three freshwater invertebrate species (Gammaridae, Ephemerellidae, and Chironomidae). The aquatic insects Diamesa sp., Drunella cryptomeria, and Gammarus suifunensis were incubated with the CNFs at the concentration of 100 mg/L during the 7-days period. Histological examination of the whole specimens and the longitudinal sections revealed no toxic effects of CNFs. However, a noticeable change in the structure of the CNFs accumulated in the intestines of the aquatic insects was found by Raman spectroscopy. The registered decrease in the relative proportion of amorphous carbon included in the CNF sample was found in the intestines of Diamesa sp. and D. cryptomeria. The registered effect can indicate a biodegradation of amorphous carbon in the digestive tract of these two insect species. In contrast, the decrease of highly structured carbons and the decrease of G-bonds intensity were registered in the digestive tract of G. suifunensis. This observation demonstrates the partial biodegradation of CNFs in the digestive tract of G. suifunensis.

Contrast-induced nephropathy in an animal model: Evaluation of novel biomarkers in blood and tissue samples.

Identification of novel biomarkers of contrast-induced nephropathy (CIN) that may more accurately detect renal function changes; reflect kidney damage; assist monitoring; and elucidate pathophysiology attract considerable scientific attention nowadays. To evaluate novel biomarkers of nephrotoxicity in blood/tissue samples of a CIN model, 10 New Zealand white rabbits were divided into group 1 (n = 5; iopromide) and group 2 (n = 5; control). Blood was drawn at 0 h (immediately), 24 h and 48 h after contrast medium (CM) administration. Animals were euthanized at 48 h and kidneys were removed. Serum creatinine (sCr)/symmetric-asymmetric dimethylarginine (SDMA-ADMA) levels were measured. CM genotoxic/cytotoxic effect was investigated 48 h post-CM exposure using micronucleus assay in lymphocytes. Cytological examination was conducted using touch preparation technique (TPT). All animals in group 1 developed CIN: mean sCr levels increased by 68.2% within 48 h. Significant SDMA-ADMA level elevation was observed at 0 h and 24 h with insignificant drop at 48 h in group 1, remaining normal in group 2 at all time-points. Significant increase in bi-nucleated cells with micronuclei and micronuclei frequency was detected in group 1. Cytokinesis block proliferation index was reduced insignificantly in group 1. TPT revealed degenerative lesions/inflammation, cell degeneration, abnormal uterine tubular casts and rubella in kidneys of all animals in group 1. Group 2 presented normal cells.

Translational Application of Circulating DNA in Oncology: Review of the Last Decades Achievements.

In recent years, the introduction of new molecular techniques in experimental and clinical settings has allowed researchers and clinicians to propose circulating-tumor DNA (ctDNA) analysis and liquid biopsy as novel promising strategies for the early diagnosis of cancer and for the definition of patients' prognosis. It was widely demonstrated that through the non-invasive analysis of ctDNA, it is possible to identify and characterize the mutational status of tumors while avoiding invasive diagnostic strategies. Although a number of studies on ctDNA in patients' samples significantly contributed to the improvement of oncology practice, some investigations generated conflicting data about the diagnostic and prognostic significance of ctDNA. Hence, to highlight the relevant achievements obtained so far in this field, a clearer description of the current methodologies used, as well as the obtained results, are strongly needed. On these bases, this review discusses the most relevant studies on ctDNA analysis in cancer, as well as the future directions and applications of liquid biopsy. In particular, special attention was paid to the early diagnosis of primary cancer, to the diagnosis of tumors with an unknown primary location, and finally to the prognosis of cancer patients. Furthermore, the current limitations of ctDNA-based approaches and possible strategies to overcome these limitations are presented.