Non-affected Tissue Research Articles

Tissue factor pathway inhibitor 2 is a potent kallikrein-related protease 12 inhibitor.

The protease activities are tightly regulated by inhibitors and dysregulation contribute to pathological processes such as cancer and inflammatory disorders. Tissue factor pathway inhibitor 2 (TFPI-2) is a serine proteases inhibitor, that mainly inhibits plasmin. This protease activated matrix metalloproteases (MMPs) and degraded extracellular matrix. Other serine proteases are implicated in these mechanisms like kallikreins (KLKs). In this study, we identified for the first time that TFPI-2 is a potent inhibitor of KLK5 and 12. Computer modeling showed that the first Kunitz domain of TFPI-2 could interact with residues of KLK12 near the catalytic triad. Furthermore, like plasmin, KLK12 was able to activate proMMP-1 and -3, with no effect on proMMP-9. Thus, the inhibition of KLK12 by TFPI-2 greatly reduced the cascade activation of these MMPs and the cleavage of cysteine-rich 61, a matrix signaling protein. Moreover, when TFPI-2 bound to extracellular matrix, its classical localisation, the KLK12 inhibition was retained. Finally, TFPI-2 was downregulated in human non-small-cell lung tumour tissue as compared with non-affected lung tissue. These data suggest that TFPI-2 is a potent inhibitor of KLK12 and could regulate matrix remodeling and cancer progression mediated by KLK12.

Club Cell Protein 16 Attenuates CD16brightCD62dim Immunosuppressive Neutrophils in Damaged Tissue upon Posttraumatic Sepsis-Induced Lung Injury.

Background Recently, identification of immunosuppressive polymorphonuclear leukocytes (PMNL) that were traditionally described as proinflammatory cells emerged in the field of posttraumatic immunity. To understand their local and remote distribution after trauma, PMNL-subsets and the impact of immunomodulatory Club Cell protein (CC)16 that correlates with pulmonary complications were assessed. Methods C57BL/6N mice were divided into three groups, receiving isolated blunt chest trauma (TxT), undergoing TxT followed by cecal ligation and puncture (CLP, TxT + CLP) after 24 h, or sham undergoing analgosedation (n = 18/group). Further, each group was subdivided into three groups receiving either no treatment (ctrl) or intratracheal neutralization of CC16 by application of anti-CC16-antibody or application of an unspecific IgG control antibody (n = 6/group). Treatment was set at the time point after TxT. Analyses followed 6 h post-CLP. PMNL were characterized via expression of CD11b, CD16, CD45, CD62L, and Ly6G by flow cytometry in bone marrow (BM), blood, spleen, lung, liver, and bronchoalveolar and peritoneal lavage fluid (BALF and PL). Apoptosis was assessed by activated (cleaved) caspase-3. Results from untreated ctrl and IgG-treated mice were statistically comparable between all corresponding sham, TxT, and TxT + CLP groups. Results Immature (CD16dimCD62Lbright) PMNL increased significantly in BM, circulation, and spleen after TxT vs. sham and were significantly attenuated in the lungs, BALF, PL, and liver. Classical-shaped (CD16brightCD62Lbright) PMNL increased after TxT vs. sham in peripheral tissue and were significantly attenuated in circulation, proposing a trauma-induced migration of mature or peripheral differentiation of circulating immature PMNL. Immunosuppressive (CD16brightCD62Ldim) PMNL decreased significantly in the lungs and spleen, while they systemically increased after TxT vs. sham. CLP in the TxT + CLP group reduced immunosuppressive PMNL in PL and increased their circulatory rate vs. isolated TxT, showing local reduction in affected tissue and their increase in nonaffected tissue. CC16 neutralization enhanced the fraction of immunosuppressive PMNL following TxT vs. sham and decreased caspase-3 in the lungs post-CLP in the TxT + CLP group, while apoptotic cells in the liver diminished post-TxT. Posttraumatic CC16 neutralization promotes the subset of immunosuppressive PMNL and antagonizes their posttraumatic distribution. Conclusion Since CC16 affects both the distribution of PMNL subsets and apoptosis in tissues after trauma, it may constitute as a novel target to beneficially shape the posttraumatic tissue microenvironment and homeostasis to improving outcomes.

Biphasic Expression of Atypical Chemokine Receptor (ACKR) 2 and ACKR4 in Colorectal Neoplasms in Association with Histopathological Findings.

Facilitating resolution of inflammation using atypical chemokine receptors (ACKR) as an anticancer strategy is considered but requires a deeper understanding of receptor role in carcinogenesis. We aimed at transcriptional analysis (RTqPCR) of ACKR2 and ACKR4 expression in colorectal adenoma-adenocarcinoma sequence in paired normal-neoplastic tissues from 96 polyps and 51 cancers. On average, ACKR2 was downregulated in neoplastic as compared to non-affected tissue in polyp (by 2.7-fold) and cancer (by 3.1-fold) patients. The maximal downregulation (by 8.2-fold) was observed in adenomas with the highest potential for malignancy and was gradually lessening through cancer stages I-IV, owing to increased receptor expression in tumors. On average, ACKR4 was significantly downregulated solely in adenocarcinomas (by 1.5-fold), less so in patients with lymph node metastasis, owing to a gradual decrease in ACKR4 expression among N0-N1-N2 cancers in non-affected tissue without changes in tumors. In adenomas, ACKR4 downregulation in neoplastic tissue increased with increasing potential for malignancy and contribution of villous growth pattern. ACKR4 expression increased in non-affected tissue with a concomitant decrease in pathological mucosa. In conclusion, the changes in ACKRs expression occur already in precancerous colorectal lesions, culminating in the adenomas with the highest potential for malignancy. Therefore, chemoprevention by manipulating ACKRs’ expression is worth exploration.

Plasma and local expressions of CircRNA CDR1as are linked with disease severity in patients with non-traumatic osteonecrosis of femoral head

ObjectiveTo investigate the correlation of plasma and local expressions of Circ CDR1as with disease severity in patients with non-traumatic osteonecrosis of femoral head (ONFH).MethodsNinety-nine non-traumatic ONFH patients receiving surgery and 99 healthy individuals were enrolled in our study. Plasma and local Circ CDR1as were detected using real-time quantitative PCR (RT-qPCR). Radiographic progression was determined using Association Research Circulation Osseous (ARCO) classification system. Harris hip score (HHS) and visual analogue scale (VAS) were used to assess the clinical severity. Receiver operating characteristic (ROC) curve was carried out to evaluate the diagnostic value of plasma Circ CDR1as with regard to the radiographic severity.ResultsPlasma Circ CDR1as expressions were significantly higher in non-traumatic ONFH patients compared with healthy controls. In non-traumatic ONFH patients, there were no significant differences of CircCDR1as expressions between patients with alcohol-induced ONFH and steroid-induced ONFH. CircCDR1as in local necrotic tissue were significantly higher than adjacent non-affected tissue. Plasma and local Circ CDR1as expressions in patients with ARCO phase 4 were markedly upregulated compared with ARCO phase 3; plasma and local Circ CDR1as expressions in patients with ARCO phase 3 were markedly upregulated compared with ARCO phase 1/2. Plasma and local CircCDR1as expressions were positively associated with ARCO classification. In addition, plasma and local Circ CDR1as expressions were positively correlated with VAS and HHS scores. ROC curve analysis indicated that plasma Circ CDR1as may act as a decent marker for radiographic progression in non-traumatic ONFH patients.ConclusionsBoth plasma and local expressions of CircRNA CDR1as are linked with disease severity in patients with non-traumatic ONFH.

Bacteriomic Profiling of Branchial Lesions Induced by Neoparamoeba perurans Challenge Reveals Commensal Dysbiosis and an Association with Tenacibaculum dicentrarchi in AGD-Affected Atlantic Salmon (Salmo salar L.).

Amoebic gill disease is a parasitic condition that commonly affects marine farmed Atlantic salmon. The causative agent, Neoparamoeba perurans, induces a marked proliferation of the gill mucosa and focal superficial necrosis upon branchial lesions. The effect that amoebic branchialitis has upon gill associated commensal bacteria is unknown. A 16S rRNA sequencing approach was employed to profile changes in bacterial community composition, within amoebic gill disease (AGD)-affected and non-affected gill tissue. The bacterial diversity of biopsies with and without diseased tissue was significantly lower in the AGD-affected fish compared to uninfected fish. Furthermore, within the AGD-affected tissue, lesions appeared to contain a significantly higher abundance of the Flavobacterium, Tenacibaculum dicentrarchi compared to adjunct unaffected tissues. Quantitative PCR specific to both N. perurans and T. dicentrarchi was used to further examine the co-abundance of these known fish pathogens. A moderate positive correlation between these pathogens was observed. Taken together, the present study sheds new light on the complex interaction between the host, parasite and bacterial communities during AGD progression. The role that T. dicentrarchi may play in this complex relationship requires further investigation.

Ефект етил 2-(хіназолін-4-іламіно)-4,5,6,7тетрагідробензо[ b ]тіофен-3-карбоксилат гідрохлориду на ДНК, РНК, білки хроматину й показники стану антиоксидантної системи в карциномі Герена та незмінених тканинах матки щурів

Стандартні підходи до вивчення специфічної фармакологічної дії та токсичності протипухлинних засобів не дають змоги провести детальну комплексну оцінку метаболічних змін поза пухлиною, хоча давно стало очевидним, що неспецифічна дія хіміотерапії може на тлі пригнічення росту пухлин викликати незворотні зміни в організмі, які надалі суттєво погіршать його шанси на видужання. Ці обставини зумовлюють актуальність включення в доклінічні дослідження нових протипухлинних засобів вивчення їхніх ефектів як на пухлини, так і на прилеглі не уражені тканини, для подальшої оптимізації хіміотерапії. Мета дослідження – порівняти вплив етил 2-(хіназолін-4-іламіно)-4,5,6,7-тетрагідробензо[b] тіофен-3-карбоксилат гідрохлориду (ЕХТК) на ДНК, РНК, білки хроматину, процеси фрагментації ДНК, вміст ТБК-реактантів і SН-груп у карциномі Герена та незмінених тканинах матки. Дослідження впливу ЕХТК проводили на білих щурах – самицях лінії Вістар. Тварини були розподілені за методом рандомізації на 3 групи по 6 тварин у кожній: 1 – інтактні щури; 2 – позитивний контроль (карцинома Герена); 3 – карцинома Герена + ЕХТК (24,37 мг/кг). Вивчення впливу ЕХТК на ДНК, РНК, білки хроматину, процеси фрагментації ДНК, вміст ТБКреактантів та небілкових і пов’язаних з білками SH-груп у карциномі Герена та незмінених тканинах матки показало, що дана сполука вірогідно змінювала більшість досліджуваних показників як порівняно з інтактними тваринами, так і з карциномою Герена без лікування. Ефект ЕХТК на фрагментацію ДНК не відзначався вузькою тканинною специфічністю. Специфічність впливу ЕХТК на клітини пухлин була найвираженішою у випадку змін вмісту ДНК, РНК, білків хроматину, а також ТБК-реактантів, небілкових і пов’язаних з білками SH-груп.

AABH (aspartyl [asparaginyl] β-hydroxylase) as a serum biomarker for breast cancer.

2 Background: Breast cancer accounts for 27% of female cancers. One in every eight women will develop breast cancer in their lifetime. More than 220,000 new cases of breast cancer will be diagnosed in the United States in 2020. Five-year survival rates for breast cancer are 98% for localized disease, 83% for regionally spread and 26% for distantly spread cancer. However, 25% of all metastases occur more than 5 years after the initial diagnosis and thus survival rates decrease as time goes out to 10 years due to disease recurrence. Current detection methods rely on imaging modalities, including mammography and MRI. No serum bio-markers have been approved for either breast cancer screening or monitoring for disease recurrence, yet. In asymptomatic women in remission from breast cancer the guidelines of the major oncologic organizations only suggest annual mammography for detection of new primary tumors. Methods: We have investigated the utility of aspartyl (asparaginyl) β-hydroxylase (AABH) as a serum bio-marker for cancer. AABH has been detected by immunohistochemical staining (IHC) in a broad range of cancers including breast cancer. It has been detected by IHC in > 99% of tumor specimens tested (n > 2000) but is absent in adjacent non-affected tissue, and in tissue samples from non-affected individuals. This led to the development of a sandwich ELISA that reliably measures AABH in serum. Results: In the current study we have utilized the assay to quantify AABH levels in the sera of patients diagnosed with Breast Cancer compared to women free of disease. Increased levels of AABH were found in the serum of 99% of patients with breast cancer (n = 189). In women not known to have cancer, AABH was essentially undetectable in serum (n = 65, specificity = 96%). Serum from several women currently in remission subsequent to treatment for breast cancer were also analyzed and found to be negative for AABH. Conclusions: Thus, measurement of serum AABH levels has great promise as a diagnostic tool for breast cancer with potential application in monitoring for disease recurrence. Elevated serum AABH in conjunction with mammography and MRI may greatly facilitate earlier diagnosis of both primary and recurrent Breast Cancer.

Impact of diffusion weighted magnetic resonance imaging in diagnosis of cervical cancer

BackgroundCervical cancer is a widespread cancer among women worldwide. In this study, we evaluated the role of diffusion weighted magnetic resonance imaging (DWI-MRI) in diagnosis of cervical carcinoma with pathological diagnosis taken as the reference. The study included 70 patients in whom cancer cervix had been suspected clinically or by transvaginal U/S and confirmed by biopsy. The control group consisted of 40 patients with a normal cervix who performed MRI for other pelvic disorders. Pelvic MR with DWI and dynamic contrast enhanced MRI (DCE-MR) were done for all patients.ResultsBoth DW-MRI and DCE-MRI revealed the same sensitivity, specificity, PPV, and accuracy; 100%, 50%, 97%, and 97%, respectively. The mean ADC value for malignant lesions was (0.82 × 10−3 ± 0.1 SD mm2/s). While the mean ADC value in the control group is (1.56 × 10–3 mm2/s). ADC value of (1.07 × 10−3 mm2/s) is a cut-off between normal cervical tissue& malignant cervical lesion with a sensitivity 97% and specificity 95.5%ConclusionEach of the (DW-MRI) and (DCE-MRI) sequences when added to the non-contrast MRI sequences in the estimation of cancer cervix had elicited the same sensitivity, specificity, and accuracy. DW–MRI was significantly beneficial in terms of diagnostic performance. For patients who cannot receive contrast medium, dynamic contrast-enhanced imaging can be replaced by DWI. The ADC value in case of cervical cancer was significantly lower than in the non-affected cervical tissue.

АЛГОРИТМ СЕГМЕНТАЦІЇ НОВОУТВОРЕНИХ ПУХЛИН НА МРТ ЗОБРАЖЕННІ ГОЛОВНОГО МОЗКУ ЗА ДОПОМОГОЮ КОМБІНАЦІЙ НЕЙРОННИХ МЕРЕЖ

The algorithm of segmentation of brain tumors in MRI images is proposed. In the iteration of the computation algorithm, the outputs of the base neural networks are used as input data for a new trained neural network, which in the future serves as a unifier in order to distinguish scar tissue or non-affected tissue from tumor cells. This approach has a complex generalization, but, thus, it is possible to improve the quality of segmentation of the tumor by a combination of neural networks. The components of the algorithm are basic classifiers that will extract complex functions of the regularities (often implicit) from the data stream, and the unifier will become a classifier that aggregates these functions. At the aggregation level, the data is derived from the classifiers, and the aggregation of the single output. When iterating the computation algorithm, the outputs of the basic classifiers are used as input data for the new trained neural network, which later acts as a unifier. The key idea of the algorithm is that the individual result for each classifier is determined based on the models previously trained, then the voxel is classified as part of the tumor if at least one of the classifiers determines it as a tumor. Further, the result of segmentation of the basic classifiers falls on the input of the already trained meta-classifier, which makes the final decision regarding the voxel's belonging to the image to the tumor cells. In this case, a special algorithm is used. The pixel algorithm proposes to classify pixels in adjacent areas based on gray levels. This method uses local information - the values of the gray levels of adjacent pixels, or, global information - the total distribution of the gray levels of adjacent pixels. The gray levels reflect the intensity of the light in each pixel. At the level of input data and manipulations with them there is an input to the input of the neural network for training.

The quantitative assessment of interstitial lung disease with positron emission tomography scanning in systemic sclerosis patients.

ObjectivesThe reversibility of interstitial lung disease (ILD) in SSc is difficult to assess by current diagnostic modalities and there is clinical need for imaging techniques that allow for treatment stratification and monitoring. 18F-Fluorodeoxyglucose (FDG) PET/CT scanning may be of interest for this purpose by detection of metabolic activity in lung tissue. This study aimed to investigate the potential role of 18F-FDG PET/CT scanning for the quantitative assessment of SSc-related active ILD.Methods 18F-FDG PET/CT scans and high resolution CT scans of eight SSc patients, including five with ILD, were analysed. For comparison, reference groups were included: eight SLE patients and four primary Sjögren’s syndrome (pSS) patients, all without ILD. A total of 22 regions of interest were drawn in each patient at apical, medial and dorsobasal lung levels. 18F-FDG uptake was measured as mean standardized uptake value (SUVmean) in each region of interest. Subsequently, basal/apical (B/A) and medial/apical (M/A) ratios were calculated at patient level (B/A-p and M/A-p) and at tissue level (B/A-t and M/A-t).ResultsSUVmean values in dorsobasal ROIs and B/A-p ratios were increased in SSc with ILD compared with SSc without ILD (P = 0.04 and P = 0.07, respectively), SLE (P = 0.003 and P = 0.002, respectively) and pSS (P = 0.03 and P = 0.02, respectively). Increased uptake in the dorsobasal lungs and increased B/A-t ratios corresponded to both ground glass and reticulation on high resolution CT.ConclusionSemi-quantitative assessment of 18F-FDG PET/CT is able to distinguish ILD from non-affected lung tissue in SSc, suggesting that it may be used as a new biomarker for SSc-ILD disease activity.

CD49b, CD87, and CD95 Are Markers for Activated Cancer-Associated Fibroblasts Whereas CD39 Marks Quiescent Normal Fibroblasts in Murine Tumor Models.

Fibroblasts are thought to be key players in the tumor microenvironment. Means to identify and isolate fibroblasts as well as an understanding of their cancer-specific features are essential to dissect their role in tumor biology. To date, the identification of cancer-associated fibroblasts is widely based on generic markers for activated fibroblasts in combination with their origin in tumor tissue. This study was focused on a deep characterization of the cell surface marker profile of cancer-associated fibroblasts in widely used mouse tumor models and defining aberrant expression profiles by comparing them to their healthy counterparts. We established a generic workflow to isolate healthy and cancer-associated fibroblasts from solid tissues, thereby reducing bias, and background noise introduced by non-target cells. We identified CD87, CD44, CD49b, CD95, and Ly-6C as cancer-associated fibroblast cell surface markers, while CD39 was identified to mark normal fibroblasts from healthy tissues. In addition, we found a functional association of most cancer-related fibroblast markers to proliferation and a systemic upregulation of CD87, and CD49b in tumor-bearing mice, even in non-affected tissues. These novel markers will facilitate the characterization of fibroblasts and shed further light in their functions and implication in cancer progression.

MicroRNA Expression Profiles in External Cervical Resorption

IntroductionExternal cervical resorption (ECR) has been challenging for its diagnosis, prevention, and treatment. Its etiology and pathogenesis are largely unknown. This study characterized microRNA (miRNA) expression patterns of human tissues from ECR lesions and identified potential messenger RNA targets and pathways. MethodsGranulomatous tissues from ECR (n = 5) and their adjacent nonaffected asymptomatic gingival connective tissues (n = 5) were collected. Similarly, chronic periodontitis (CP) and control samples were collected (n = 3). Quantitative reverse transcription polymerase chain reaction array analysis compared the expression profiles of 88 miRNAs between diseases. Differentially expressed miRNAs were identified using the Student t test. Bioinformatics for messenger RNA (miRWalk) and KEGG pathway analyses were performed to identify predicted target genes and biological/cellular functions and signaling pathways. ResultsThree miRNAs (miR-20a-5p, miR-210-3p, and miR-99a-4p) were significantly down-regulated and 1 miRNA (miR-122-5p) was significantly up-regulated in ECR (P < .05). One up-regulated and 1 down-regulated miRNA reached the significance threshold in CP. A comparison of miRNA expression in ECR and CP identified 3 differentially expressed miRNAs, indicating differences in disease pathobiology. Inflammation-associated Wnt, PI3K-Akt, mitogen-activated protein kinases signaling, and bone formation–associated transforming growth factor beta pathways were identified and predicted to be modulated by differentially expressed miRNAs in both ECR and CP. Biological processes unique to each disease entity were identified, such as T- and B-cell receptor signaling pathways, osteoclast differentiation, and extracellular matrix–receptor interaction for CP. Glycosaminoglycan biosynthesis, mineral absorption, and insulin signaling pathways for ECR were identified. ConclusionsThis proof-of-principle in vivo study indicated that ECR has both common and unique miRNA expression profiles in comparison with CP, which are predicted to target genes regulating inflammation, immunity, and metabolism of mineralized tissues.

Schematic algorithm for surgical treatment of idiopathic granulomatous mastitis using combined steroids and therapeutic mammoplasty techniques

Introduction Idiopathic granulomatous mastitis (IGM) is a rare, benign, chronic, inflammatory lesion of the breast of unknown etiology. Clinically, it can be mistaken for inflammatory and neoplastic disorders of the breast. Laboratory investigations with culture and sensitivity, PCR for Mycobacterium tuberculosis and core needle biopsy are crucial. Treatment is controversial, but oral steroids are usually started with, followed by surgical resection if there is no complete response. Aim This study discusses the use of therapeutic mammoplasty techniques to excise the mass and preserve breast aesthetic appearance concerning patient satisfaction and recurrence. Patients and methods Fifty IGM patients were included with moderate to large breasts with masses 20–50% of the breast size after failure of medical treatment with prednisolone, or occurrence of therapy-related complications. Preoperative core needle biopsy, smear for Ziehl–Nelseen acid fast staining, and culture and sensitivity test. Mammography and ultrasonography were done. An informed consent was obtained regarding the operative procedure and research. On surgery we excise the inflammatory mass with a rim of normal nonaffected tissues around with most of retroareolar duct system. Patients were followed up for 1 year in regular outpatient visits, where recurrence and patients’ satisfaction were assessed. Results The most important postoperative complication was disease recurrence (4%) and deformity (10%). The most common one was minor wound dehiscence (40%). For patient satisfaction, the median mean percent satisfaction score was 74.50%. Conclusion The use of therapeutic mammoplasty in surgical management of IGM has low recurrence rate and high postoperative patient satisfaction.

Uptake in non-affected bone tissue does not differ between [18F]-DCFPyL and [68Ga]-HBED-CC PSMA PET/CT.

Introduction[68Ga]PSMA-HBED-CC and [18F]DCFPyL show a high potential for the detection of recurrent prostate cancer. While 18F-based tracers have several advantages in availability and image resolution, their sensitivity in the skeleton might be impaired by released [18F]fluoride due to its high bone affinity. In turn, chemically unbound trivalent 68Ga might also accumulate in osseous tissue, in cases of occupied binding sites of plasma proteins and thereby influence bone signal.MethodsA comparison of average bone SUV was performed in 17 bone-negative and 4 bone-positive patients. All patients underwent PET/CT 125 minutes after application of [18F]DCFPyL and 73 minutes after application of [68Ga]PSMA-HBED-CC at another date.ResultsNative SUVs in unaffected bone tissue and SUVs relative to liver uptake were lower in [18F]DCFPyL (0.49) than in [68Ga]PSMA-HBED-CC scans (0.52). SUVs relative to gluteal muscles did not differ between the two tracers. Average lesional SUVs did not differ between tracers.ConclusionNo difference of average bone signal intensity was observed for [18F]DCFPyL-PET/CT in comparison to [68Ga]PSMA-HBED-CC scans indicating that diagnostic assessment of the skeleton is not affected by non-specific accumulation of free [18F]fluoride or 68Ga.

Innovations in molecular biology and proteomics adapted to in vivo peptide phage display define active protein domains as biomarkers in multiple sclerosis neuroinflammation

With the double goal to streamline biomarker discovery of Multiple Sclerosis neuroinflammatory pathology entangled by non-affected Central Nervous System tissue and to better characterize the complex molecular interactions at the altered blood brain barrier (BBB) that becomes permissive to extravasation of blood derived immune cells and compounds, we choose to identify peptide ligands reacting with inflammatory CNS lesions. With no “a-priori” for targets by applying in vivo the phage displayed 12aa peptide screening in CNS neuroinflammation of the experimental autoimmune encephalomyelitis (EAE) rat model of MS and healthy controls, we generated two complex phage displayed peptide repertoires. By developing molecular DNA subtraction of both phage repertoires we generated a subtraction phage repertoire of EAE specific peptides. In a bioinformatics approach aligning the 270,612 EAE peptides to the human proteome the formed peptide clusters allowed the ranking of proteins being potential “mimicked” indicators of functional protein interactions. Experimentally, among the most high score mimicked proteins, derived synthesized short peptides (22aa) of mimicked domains were tested for labeling of BBB alterations in CNS sections of MS and other chronic neurodegenerative diseases. In vitro studies using the BBB model hCMEC/D3 confirmed their binding to human endothelial cells under proinflammatory stimulation. Finally, with a derived peptide covalently linked to a nanocarrier contrast agent, we could target in vivo and monitor by MRI, experimentally induced inflammatory CNS lesions. Our strategy of combining the developed molecular biology and bioinformatics approaches adapted to phage display is applicable to many disease models to identify biomarkers and functional proteins/domains.

Phacomatosis Pigmentokeratotica

AbstractIn the group of the epidermal nevus syndromes, Happle defined in 1996 a separate entity characterized by the presence of an organoid epidermal nevus, sometimes showing sebaceous differentiation, and a speckled lentiginous nevus of the papular type, occasionally associated with extracutaneous anomalies including neurological, ophthalmological, and skeletal abnormalities. In particular, the syndrome is associated with mental retardation, epilepsy, deafness, hemiatrophy, dysesthesia, and hyperhidrosis, strabismus, lipodermoid of conjunctiva, coloboma and ptosis, and kyphosis, scoliosis, limb asymmetry, and hypertrophy. Rarely, hypertension, vascular abnormalities, atrioventricular block, hypophosphatemic rickets, and pheochromocytoma may occur. The organoid nevus follows the lines of Blaschko whereas the speckled lentiginous nevus is arranged in a checkerboard pattern. For this syndrome, the term “Phacomatosis Pigmentokeratotica” has been coined and, at the present, it is considered a very rare clinical entity, with less than 20 cases reported in the literature. Recent genetic findings have included this syndrome in the group of the mosaic RASopathies, after the discovery of mutations in the HRAS gene occurring in both sebaceous and vascular nevi, but not in nonaffected tissues.

Abstract 1470: Relative telomere length in tumor tissue and adjacent mucosa of colorectal cancer patients

Abstract Colorectal cancer (CRC) represents a major public health problem, being the fourth leading cause of cancer related deaths mainly due to the late diagnosis and a lack of proper prognostic markers. According to the current knowledge, shortening of telomeres is important contributor in cancer onset. Telomeres, repetitive TTAGGG sequences and boud proteins, are structures at the end of all linear eukaryotic chromosomes. Telomeres are double-edged sword regarding to cancer. On one hand, they serve as a potent tumor-supressor mechanism limiting the cells ability of uncontroled growth. However, on the other hand, shortening of telomeres can lead to the formation of anaphase bridges, initiation of breakage-fusion-bridges cycles and eventually to the induction of chromosome instability phenotype. Telomeric regions are composed of microsatellite repeats and DNA mismatch repair (MMR) defects should directly affect telomere length. Studies looking into the relationship between microsatellite instability (MSI) and telomere length have been sparse. We hypothesize that CRC patients with MSI should have better prognosis, longer survival, also due to the MMR defects in telomere regions replication failure. The aim of our present study was to investigate relative telomere length (RTL) in tumor tissue and adjacent mucosa of 650 CRC patients; it is the largest study on RTL in target tissue so far. Moreover, we measured RTL in peripheral blood lymphocytes obtained from 150 cancer patients. Tumor and adjacent non-affected tissue samples were obtained from patients at surgical resection. RTL was measured using monochrome multiplex quantitative PCR method. Despite the analysis is still running, our preliminary results showed significantly shorter RTL in tumor tissues as compared to adjacent mucosa. The shortest RTL was discovered in right, ascendent part of colon (0.68±0.59, 25.5% of cases), followed by left, descendent colon (0.77±0.47, 37.6%) and the longest RTL was in rectal tumors (0.92±0.69, 36.9%). We recorded gradually increasing RTL ratio (RTL in tumor tissue/ RTL in adjacent mucosa) in later stages according to TNM staging system. Interestingly, patients with RTL ratio &amp;gt; 1 had also worse survival. RTL of CRC tissue and adjacent mucosa may provide a valuable prognostic tool in patients treatment management. This study was supported by GACR 15-14789S and by the National Sustainability Program I (NPU I) Nr. LO1503 provided by the Ministry of Education Youth and Sports of the Czech Republic Citation Format: Michal Kroupa, Vaclav Liska, Krishna Rachakonda, Marketa Urbanova, Michaela Schneiderova, Katerina Jiraskova, Ondrej Vycital, Ludmila Vodickova, Rajiv Kumar, Pavel Vodicka. Relative telomere length in tumor tissue and adjacent mucosa of colorectal cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1470.

Three Cases of Exclusively Extragenital Canine Transmissible Venereal Tumor (cTVT)

Background: Canine Transmissible Venereal Tumor (cTVT) is a neoplasia that affects mainly the genital organs of dogs, but can rich extragenital sites as well. It´s a tumor characterized microscopically by the presence of vacuolized round cells. Transmission occurs by implantation of these cells in non-affected tissues and the treatment is based on vincristine chemotherapy.Cases: Case 1. A 5-year-old intact male Poodle, presenting an increase volume of nasal plane came for veterinary care at a private veterinary clinic. The animal had bilateral bloody nasal secretion and dyspnea. The external genitalia had no alterations. The cytological evaluation confirmed cTVT. Treatment with vincristine sulfate weekly showed a rapid responsewith improvement of the respiratory condition, total remission of the mass and absence of neoplastic cells in cytology. Case 2. A 5-year-old mixed-breed canine bitch, weighing 6.7 kg, was brought to the State University of Santa Cruz Veterinary Hospital (UESC-VH), showing an increase volume in the nasal plan region, with complaints about sneezing, nasal bleeding,respiratory distress with approximately 4 months of evolution. The owner informed that the mother of these female dog, that lived in the same environment, died a month before the beginning of clinical signs of the bitch of this case, and showed a reddish vaginal mass with intense bleeding. Intranasal exfoliative cytology showed moderately cellular sample compatible with cTVT. The treatment with vincristine sulphate for 6 weeks, showed completely remission of all clinical signs. Case 3. A 3-year-old mixed-breed male dog was brought to the UESC-VH with a reddish, friable mass located in the left eye. The citology confirmed the clinical suspicion of cTVT. After six weekly sessions of chemotherapy with vincristine sulfate, the tumor regressed and a new cytological evaluation was performed, without visible of tumor cells. By the end of the treatment, the dog was diagnosed with phitisis bulbi, and one year later, due to recurrent ulcerative keratitis, the enucleation was performed and the histopathological examination of the eye did not identify the presence of tumor cells.Discussion: Two of the dogs cited in this report had freely streets access, without supervision of the owners, and they are likely to have contracted cTVT on one of those occasions. The animal’s care style acts as a risk factor for the development of neoplasia. Regarding the third animal, the close contact with another female dog, who had compatible vaginal cTVTclinical signs was probably the factor that determined the transmission. None of the animals cited in this report had lesions on their external genitalia. The extragenital presentation may be attributed to the social behavior of licking and sniffing the genitalia of carrier animals, which may lead to the natural implantation of the viable cells of the cTVT into the ocular and nasal mucosa. About the clinical signs manifested, in the cases of involvement of the nasal structures, the main signs described in literature are bloody nasal secretion, sneezing, dyspnea and increased nasal plane volume, and are similar to those observed in the animals cited in this report. In the case of ocular cTVT, the increase volume with impairment and deformity of all ocular structures, as well as pain and pruritus corroborate with the clinical findings observed in the literature. The cytopathological test was the diagnostic tool used in all cases cited in this report and the cytopathological findings corroborates with those described in the literature. Vincristine sulfate is the drug of choice for the treatment of cTVT cases, and in the dogs of this report, this drug was successfully used leading to complete remission of lesions and clinical signs, as observed in other studies.Keywords: nasal mucosa, ocular mucosa, venereal disease, round cells.

Role of Diffusion Weighted MRI in Diagnosis of Cervical Cancer

Background: Diffusion weighted magnetic resonance imaging (DW-MRI) is a functional, non-invasive imaging technique which generates tissue contrast from differences in mobility of water molecules that occurs during an MR pulse sequence. Information regarding the integrity of cellular membranes and tissue cellularity can be obtained, so that DW-MRI can now be included in routine patient assessment. Aim of the Work: The aim of this study is to evaluate the role of DW-MRI in the diagnosis of cervical carcinoma, withpathological diagnosis was taken as the reference. Patients and Methods: This is a retrospective study that included 20 patients in whom cervical cancer had been suspected clinically or by transvaginal ultrasound (U/S) and the control group consisted of 20 patients in whom cervical cancer had not been suspected and MRI was performed because of other Pelvic diseases. The study was conducted in El-Demerdash Hospital. The patients were referred from the Gynecology Department to the Radiology Department (Women’s imaging unit) for further MRI assessment with diffusion weighted images (DWIs). Results: Lesions in all cases show restricted diffusion, however on apparent diffusion coefficient (ADC) map, only one case showed high signal proved to be chronic cervicitis on histopathology. Also, the mean ADC values for malignant lesions were (0.82 x10-3 mm2/sec), while the mean ADC value in the control group was (1.58x10–3 mm2/sec). Therefore, ADC value of (1.04 x10-3 mm2/sec) is a cut off between normal cervical tissue and malignant cervical lesion by sensitivity 95% and specificity 95 %. DWIs had elicited the same accuracy to Dynamic contrast-enhanced (DCE) sequences (95%) when added to the non-contrast MRI in the estimation of cancer cervix. Conclusion: Our results proved that (DW–MRI) was significantly beneficial in terms of diagnostic performance that increases the radiologist’s confidence in image interpretation. So, it implies a non-invasive technique which can be used especially if contrast intake is avoided as in pregnancy. Thus, we suggest that DWI should be included in the routine pelvic MRI protocol. The ADC value in case of cervical carcinoma was significantly lower than in the normal cervical tissue. The ADC threshold of (1.04 x10-3 mm2/sec) was a cut off value, which was detected when differentiating between cancer-affected and non-affected cervical tissues

Expression Patterns of CXCL12 and its Receptor in Colorectal Carcinoma.

Stromal cell-derived factor-1 (also called CXCL12) and its receptor, CXCR4, have a key role in the pathogenesis and tumorigenesis of various cancers. The aims of the current study were to quantitatively examine the expression of CXCR4 and CXCL12 genes in colorectal cancer and to correlate their expression degree with clinicopathological features. Tumor tissue samples were collected from 47 patients with CRC. Total RNA was isolated from resection tissues and real-time PCR analysis was performed to examine mRNA levels of CXCL12 and CXCR4 genes. No significant differences were observed for both CXCL12 and CXCR4 between tumor tissues and the adjacent non-affected tissues, although a borderline significant correlation (p = 0.052) were detected between gene expression of CXCL12 and CXCR4 in tumor tissues. Our results also indicated that there was no significant correlation between expression pattern of CXCL12/CXCR4 and clinicopathological variables. Our data showed that CXCL12 and CXCR4 are expressed simultaneously in colorectal carcinoma tissues, suggesting that expression of these chemokines and corresponding receptors may play a pivotal role in colorectal tumorigenesis, although it cannot be as a predictive factor for disease progression.