CCAT1 lncRNA is chromatin-retained and post-transcriptionally spliced.

Super-enhancers are unique gene expression regulators widely involved in cancer development. Spread over large DNA segments, they tend to be found next to oncogenes. The super-enhancer c-MYC locus forms long-range chromatin looping with nearby genes, which brings the enhancer and the genes into proximity, to promote gene activation. The colon cancer-associated transcript 1 (CCAT1) gene, which is part of the MYC locus, transcribes a lncRNA that is overexpressed in colon cancer cells through activation by MYC. Comparing different types of cancer cell lines using RNA fluorescence in situ hybridization (RNA FISH), we detected very prominent CCAT1 expression in HeLa cells, observed as several large CCAT1 nuclear foci. We found that dozens of CCAT1 transcripts accumulate on the gene locus, in addition to active transcription occurring from the gene. The accumulating transcripts are released from the chromatin during cell division. Examination of CCAT1 lncRNA expression patterns on the single-RNA level showed that unspliced CCAT1 transcripts are released from the gene into the nucleoplasm. Most of these unspliced transcripts were observed in proximity to the active gene but were not associated with nuclear speckles in which unspliced RNAs usually accumulate. At larger distances from the gene, the CCAT1 transcripts appeared spliced, implying that most CCAT1 transcripts undergo post-transcriptional splicing in the zone of the active gene. Finally, we show that unspliced CCAT1 transcripts can be detected in the cytoplasm during splicing inhibition, which suggests that there are several CCAT1 variants, spliced and unspliced, that the cell can recognize as suitable for export.

The non-coding RNAs of the H19-IGF2 imprinted loci: a focus on biological roles and therapeutic potential in Lung Cancer.

Since it was first described, the imprinted cluster 11p15.5 has been reported to be deregulated in a variety of pediatric and adult cancers including that of the lung. Both protein coding and non-coding genes functioning as oncogenes or as tumor suppressor genes reside within this cluster. Oncomirs that can function as oncogenes or as tumor suppressors have also been reported. While a complete account of the role played by the 11p15.5 imprinted cluster in lung cancer is beyond the scope of this review, we will focus on the role of the non-coding RNAs processed from the H19-IGF2 loci. A special emphasis will be given to the H19/miR-675 gene locus. Their potential diagnostic and therapeutic use in lung cancer will be described.

Differential expression of colon cancer associated transcript1 (CCAT1) along the colonic adenoma-carcinoma sequence.

BACKGROUND: The transition from normal epithelium to adenoma and, to invasive carcinoma in the human colon is associated with acquired molecular events taking 5-10 years for malignant transformation. We discovered CCAT1, a non-coding RNA over-expressed in colon cancer (CC), but not in normal tissues, thereby making it a potential disease-specific biomarker. We aimed to define and validate CCAT1 as a CC-specific biomarker, and to study CCAT1 expression across the adenoma-carcinoma sequence of CC tumorigenesis. METHODS: Tissue samples were obtained from patients undergoing resection for colonic adenoma(s) or carcinoma. Normal colonic tissue (n = 10), adenomatous polyps (n = 18), primary tumor tissue (n = 22), normal mucosa adjacent to primary tumor (n = 16), and lymph node(s) (n = 20), liver (n = 8), and peritoneal metastases (n = 19) were studied. RNA was extracted from all tissue samples, and CCAT1 expression was analyzed using quantitative real time-PCR (qRT-PCR) with confirmatory in-situ hybridization (ISH). RESULTS: Borderline expression of CCAT1 was identified in normal tissue obtained from patients with benign conditions [mean Relative Quantity (RQ) = 5.9]. Significant relative CCAT1 up-regulation was observed in adenomatous polyps (RQ = 178.6 ± 157.0; p = 0.0012); primary tumor tissue (RQ = 64.9 ± 56.9; p = 0.0048); normal mucosa adjacent to primary tumor (RQ = 17.7 ± 21.5; p = 0.09); lymph node, liver and peritoneal metastases (RQ = 11,414.5 ± 12,672.9; 119.2 ± 138.9; 816.3 ± 2,736.1; p = 0.0001, respectively). qRT-PCR results were confirmed by ISH, demonstrating significant correlation between CCAT1 up-regulation measured using these two methods. CONCLUSION: CCAT1 is up-regulated across the colon adenoma-carcinoma sequence. This up-regulation is evident in pre-malignant conditions and through all disease stages, including advanced metastatic disease suggesting a role in both tumorigenesis and the metastatic process.

Differentiating benign from malignant thyroid nodules using micro ribonucleic acid amplification in residual cells obtained by fine needle aspiration biopsy.

BACKGROUND: Fine needle aspiration biopsy (FNAB) is the most commonly used diagnostic tool to differentiate benign from malignant thyroid nodules. Nevertheless, some FNAB cytology results are not definite. In such cases diagnostic thyroid lobectomy is performed with malignancy rate on final histopathology ranging at 15%-75%. The aim of this study was to improve on the accuracy of FNAB-based cytology by amplification of microRNAs (micro ribonucleic acids [miRs]) from the residual cells left in the FNAB needle after submission for cytology. METHODS: Residual cells were collected from the needle cup after FNAB cytology of 77 consecutive patients with thyroid nodules. miR-enriched RNA was extracted for all patients with cytology showing either follicular lesion or suspicion for malignancy (n=11). The expression of miR-21, -31, -146b, -187, -221, and -222 was determined using real-time polymerase chain reaction. Results were compared with final surgical histopathology. RESULTS: RNA was successfully extracted from all FNAB specimens. Five patients had FNAB cytology suspicious for malignancy. The miR panel was positive in all five (100%). Six patients had follicular lesions on FNAB. The miR panel was positive in three of four patients (75%) with confirmed malignancy and was negative in two of two (0%) patients with benign pathology results. This corresponded to a specificity of 100%, sensitivity of 88%, and accuracy of 90%. CONCLUSIONS: RNA extraction from FNAB residual cells is feasible, and a miR panel amplified from the extracted RNA seems like a promising diagnostic tool in this limited number of patients.

Colon cancer associated transcript-1: a novel RNA expressed in malignant and pre-malignant human tissues.

Early detection of colorectal cancer (CRC) is currently based on fecal occult blood testing (FOBT) and colonoscopy, both which can significantly reduce CRC-related mortality. However, FOBT has low-sensitivity and specificity, whereas colonoscopy is labor- and cost-intensive. Therefore, the discovery of novel biomarkers that can be used for improved CRC screening, diagnosis, staging and as targets for novel therapies is of utmost importance. To identify novel CRC biomarkers we utilized representational difference analysis (RDA) and characterized a colon cancer associated transcript (CCAT1), demonstrating consistently strong expression in adenocarcinoma of the colon, while being largely undetectable in normal human tissues (p < 000.1). CCAT1 levels in CRC are on average 235-fold higher than those found in normal mucosa. Importantly, CCAT1 is strongly expressed in tissues representing the early phase of tumorigenesis: in adenomatous polyps and in tumor-proximal colonic epithelium, as well as in later stages of the disease (liver metastasis, for example). In CRC-associated lymph nodes, CCAT1 overexpression is detectable in all H&E positive, and 40.0% of H&E and immunohistochemistry negative lymph nodes, suggesting very high sensitivity. CCAT1 is also overexpressed in 40.0% of peripheral blood samples of patients with CRC but not in healthy controls. CCAT1 is therefore a highly specific and readily detectable marker for CRC and tumor-associated tissues.

Detection of endogenous K-ras mRNA in living cells at a single base resolution by a PNA molecular beacon.

Detection of mRNA alterations is a promising approach for identifying biomarkers as means of differentiating benign from malignant lesions. By choosing the KRAS oncogene as a target gene, two types of molecular beacons (MBs) based on either phosphothioated DNA (PS-DNA-MB) or peptide nucleic acid (TO-PNA-MB, where TO = thiazole orange) were synthesized and compared in vitro and in vivo. Their specificity was examined in wild-type KRAS (HT29) or codon 12 point mutation (Panc-1, SW480) cells. Incubation of both beacons with total RNA extracted from the Panc-1 cell line (fully complementary sequence) showed a fluorescent signal for both beacons. Major differences were observed, however, for single mismatch mRNA transcripts in cell lines HT29 and SW480. PS-DNA-MB weakly discriminated such single mismatches in comparison to TO-PNA-MB, which was profoundly more sensitive. Cell transfection of TO-PNA-MB with the aid of PEI resulted in fluorescence in cells expressing the fully complementary RNA transcript (Panc-1) but undetectable fluorescence in cells expressing the K-ras mRNA that has a single mismatch to the designed TO-PNA-MB (HT29). A weaker fluorescent signal was also detected in SW480 cells; however, these cells express approximately one-fifth of the target mRNA of the designed TO-PNA-MB. In contrast, PS-DNA-MB showed no fluorescence in all cell lines tested post PEI transfection. Based on the fast hybridization kinetics and on the single mismatch discrimination found for TO-PNA-MB we believe that such molecular beacons are promising for in vivo real-time imaging of endogenous mRNA with single nucleotide polymorphism (SNP) resolution.

A conventional-to-spectral CT image translation augmentation workflow for robust contrast injection-independent organ segmentation.

PURPOSE: In computed tomography (CT) cardiovascular imaging, the numerous contrast injection protocols used to enhance structures make it difficult to gather training datasets for deep learning applications supporting diverse protocols. Moreover, creating annotations on noncontrast scans is extremely tedious. Recently, spectral CT's virtual-noncontrast images (VNC) have been used as data augmentation to train segmentation networks performing on enhanced and true-noncontrast (TNC) scans alike, while improving results on protocols absent of their training dataset. However, spectral data are not widely available, making it difficult to gather specific datasets for each task. As a solution, we present a data augmentation workflow based on a trained image translation network, to bring spectral-like augmentation to any conventional CT dataset. METHOD: The conventional CT-to-spectral image translation network (HUSpectNet) was first trained to generate VNC from conventional housnfied units images (HU), using an unannotated spectral dataset of 1830 patients. It was then tested on a second dataset of 300 spectral CT scans by comparing VNC generated through deep learning (VNCDL ) to their true counterparts. To illustrate and compare our workflow's efficiency with true spectral augmentation, HUSpectNet was applied to a third dataset of 112 spectral scans to generate VNCDL along HU and VNC images. Three different three-dimensional (3D) networks (U-Net, X-Net, and U-Net++) were trained for multilabel heart segmentation, following four augmentation strategies. As baselines, trainings were performed on contrasted images without (HUonly) and with conventional gray-values augmentation (HUaug). Then, the same networks were trained using a proportion of contrasted and VNC/VNCDL images (TrueSpec/GenSpec). Each training strategy applied to each architecture was evaluated using Dice coefficients on a fourth multicentric multivendor single-energy CT dataset of 121 patients, including different contrast injection protocols and unenhanced scans. The U-Net++ results were further explored with distance metrics on every label. RESULTS: Tested on 300 full scans, our HUSpectNet translation network shows a mean absolute error of 6.70 ± 2.83 HU between VNCDL and VNC, while peak signal-to-noise ratio reaches 43.89 dB. GenSpec and TrueSpec show very close results regardless of the protocol and used architecture: mean Dice coefficients (DSCmean ) are equal with a margin of 0.006, ranging from 0.879 to 0.938. Their performances significantly increase on TNC scans (p-values < 0.017 for all architectures) compared to HUonly and HUaug, with DSCmean of 0.448/0.770/0.879/0.885 for HUonly/HUaug/TrueSpec/GenSpec using the U-Net++ architecture. Significant improvements are also noted for all architectures on chest-abdominal-pelvic scans (p-values < 0.007) compared to HUonly and for pulmonary embolism scans (p-values < 0.039) compared to HUaug. Using U-Net++, DSCmean reaches 0.892/0.901/0.903 for HUonly/TrueSpec/GenSpec on pulmonary embolism scans and 0.872/0.896/0.896 for HUonly/TrueSpec/GenSpec on chest-abdominal-pelvic scans. CONCLUSION: Using the proposed workflow, we trained versatile heart segmentation networks on a dataset of conventional enhanced CT scans, providing robust predictions on both enhanced scans with different contrast injection protocols and TNC scans. The performances obtained were not significantly inferior to training the model on a genuine spectral CT dataset, regardless of the architecture implemented. Using a general-purpose conventional-to-spectral CT translation network as data augmentation could therefore contribute to reducing data collection and annotation requirements for machine learning-based CT studies, while extending their range of application.

Spectral augmentation for heart chambers segmentation on conventional contrasted and unenhanced CT scans: an in-depth study.

PURPOSE: Recently, machine learning has outperformed established tools for automated segmentation in medical imaging. However, segmentation of cardiac chambers still proves challenging due to the variety of contrast agent injection protocols used in clinical practice, inducing disparities of contrast between cavities. Hence, training a generalist network requires large training datasets representative of these protocols. Furthermore, segmentation on unenhanced CT scans is further hindered by the challenge of obtaining ground truths from these images. Newly available spectral CT scanners allow innovative image reconstructions such as virtual non-contrast (VNC) imaging, mimicking non-contrasted conventional CT studies from a contrasted scan. Recent publications have demonstrated that networks can be trained using VNC to segment contrasted and unenhanced conventional CT scans to reduce annotated data requirements and the need for annotations on unenhanced scans. We propose an extensive evaluation of this statement. METHOD: We undertake multiple trainings of a 3D multi-label heart segmentation network with (HU-VNC) and without (HUonly) VNC as augmentation, using decreasing training dataset sizes (114, 76, 57, 38, 29, 19 patients). At each step, both networks are tested on a multi-vendor, multi-centric dataset of 122 patients, including different protocols: pulmonary embolism (PE), chest-abdomen-pelvis (CAP), heart CT angiography (CTA) and true non-contrast scans (TNC). An in-depth comparison of resulting Dice coefficients and distance metrics is performed for the networks trained on the largest dataset. RESULTS: HU-VNC-trained on 57 patients significantly outperforms HUonly trained on 114 regarding CAP and TNC scans (mean Dice coefficients of 0.881/0.835 and 0.882/0.416, respectively). When trained on the largest dataset, significant improvements in all labels are noted for TNC and CAP scans (mean Dice coefficient of 0.882/0.416 and 0.891/0.835, respectively). CONCLUSION: Adding VNC images as training augmentation allows the network to perform on unenhanced scans and improves segmentations on other imaging protocols, while using a reduced training dataset.

Combination of Helicobacter pylori strain and tumor necrosis factor-alpha polymorphism of the host increases the risk of peptic ulcer disease in children.

BACKGROUND: Helicobacter pylori infection is probably acquired in childhood and causes a vigorous immune response. It is unclear why only a subgroup of infected children develops peptic ulcer disease. We have previously reported that iceA1 strains tend to be associated with duodenal disease in children. However, the pathogenesis probably does not depend solely on the H pylori strain but also on the variability of the host response. OBJECTIVES: The aim of this study was to assess the significance of tumor necrosis factor-alpha (TNF-alpha) promoter polymorphism in relation to infection with H pylori strains in children. METHODS: A total of 113 antral biopsies of H pylori-positive children (ages 2-18 years) were analyzed. Of these, 23 had duodenal disease, including erosive duodenitis and/or duodenal ulceration, and 90 had gastritis only. H pylori infection was diagnosed by bacterial culture and histology. Patient genomic DNA extracted from the antral biopsy was used to characterize the genetic polymorphism of TNF-alpha promoter at nucleotide positions -308 and -238 by polymerase chain reaction-based restriction fragment-length polymorphism. All H pylori strains were examined for cytotoxin-associated gene A and induced-by-contact-with-epithelium gene (iceA1). RESULTS: A total of 31% of children with duodenal disease were infected with iceA1 positive strains and had the -238 G to A polymorphism in the TNF-alpha gene vs only 1.6% of children with gastritis alone (P < 0.0005). CONCLUSIONS: The combination of bacterial iceA1 and TNF-alpha 238 G to A polymorphism may be a risk factor for peptic ulcer disease in children infected with H pylori. Larger studies are needed to confirm this association.

The role of the oncofetal H19 lncRNA in tumor metastasis: orchestrating the EMT-MET decision.

Long non-coding RNA (lncRNA) genes are emerging as key players in the metastatic cascade. Current evidence indicate that H19 lncRNA and the microRNA(miRNA) miR-675, which is processed from it, play crucial roles in metastasis, through the regulation of critical events specifically the epithelial to mesenchymal (EMT) and the mesenchymal to epithelial transitions (MET). This review summarizes recent mechanistic pathways and tries to put together seemingly conflicting data from different reports under one proposed general scheme underlying the various roles of H19/miR-675 in the metastatic cascade. We propose several approaches to harnessing this knowledge for translational medicine.

Incidence and clinical manifestations of breast milk-acquired Cytomegalovirus infection in low birth weight infants.

OBJECTIVES: To determine the incidence and clinical manifestations of human breast milk (HMB)-associated acquired cytomegalovirus (CMV) infection in small premature infants. STUDY DESIGN: A prospective study of premature infants born at or prior to 32 weeks gestation, and or infants weighing 1500 g or less at birth. The babies were divided into two groups: Group 1 included babies of CMV seropositive mothers who received HBM throughout the study period. Group 2 included babies of seronegative mothers or babies that did not receive HBM at all. Urine sample were obtained once weekly from birth until the age of 8 weeks or until discharge and were tested for the presence of CMV-DNA by PCR. RESULTS: Four of 70 infants from group 1 (5.7%, 95% CI, 0 to 11%) acquired CMV infection between the ages of 3 and 7 weeks as compared to none of 26 babies in group 2. Only one infected baby had severe CMV disease with complete recovery. CONCLUSION: The relative incidence of HBM-associated CMV infection and the severity of HBM-associated CMV disease in premature infants are low.

Colon Cancer Associated Transcript-1 (CCAT1) Expression in Adenocarcinoma of the Stomach.

BACKGROUND: Long non-coding RNAs (lncRNAs) have been shown to have functional roles in cancer biology and are dys-regulated in many tumors. Colon Cancer Associated Transcript -1 (CCAT1) is a lncRNA, previously shown to be significantly up-regulated in colon cancer. The aim of this study is to determine expression levels of CCAT1 in gastric carcinoma (GC). METHODS: Tissue samples were obtained from patients undergoing resection for gastric carcinoma (n=19). For each patient, tumor tissue and normal appearing gastric mucosa were taken. Normal gastric tissues obtained from morbidly obese patients, undergoing laparoscopic sleeve gastrectomy served as normal controls (n=19). A human gastric carcinoma cell line (AGS) served as positive control. RNA was extracted from all tissue samples and CCAT1 expression was analyzed using quantitative real time-PCR (qRT-PCR). RESULTS: Low expression of CCAT1 was identified in normal gastric mucosa samples obtained from morbidly obese patients [mean Relative Quantity (RQ) = 1.95±0.4]. AGS human gastric carcinoma cell line showed an elevated level of CCAT1 expression (RQ=8.02). Expression levels of CCAT1 were approximately 10.8 fold higher in GC samples than in samples taken from the negative control group (RQ=21.1±5 vs. RQ=1.95±0.4, respectively, p<0.001). Interestingly, CCAT1 expression was significantly overexpressed in adjacent normal tissues when compared to the negative control group (RQ = 15.25±2 vs. RQ=1.95±0.4, respectively, p<0.001). Tissues obtained from recurrent GC cases showed the highest expression levels (RQ = 88.8±31; p<0.001). Expression levels increased with tumor stage (T4- 36.4±15, T3- 16.1±6, T2- 4.7±1), however this did not reach statistical significance (p=0.2). There was no difference in CCAT1 expression between intestinal and diffuse type GC (RQ=22.4±7 vs. 22.4±16, respectively, p=0.9). Within the normal gastric tissue samples, no significant difference in CCAT1 expression was observed in helicobacter pylori negative and positive patients (RQ= 2.4±0.9 vs. 0.93±0.2, respectively, p=0.13). CONCLUSION: CCAT1 is up-regulated in gastric cancer, and may serve as a potential bio-marker for early detection and surveillance.

A single Mediterranean, possibly Jewish, origin for the Val59Gly CDKN2A mutation in four melanoma-prone families.

We have screened for CDKN2A germline mutations in 49 Jewish families with two or more cases of melanoma. The Val59Gly mutation, one of the three different alterations identified among these families, was also detected independently in two kindreds from France and one from Spain. The impact of the Val59Gly substitution on the function of the cyclin-dependent kinase inhibitor p16(INK4a), a product of the CDKN2A gene, was assessed by protein-protein interaction and cell proliferation assays and related to potential structural alterations predicted by molecular modeling. Seven microsatellite markers in the vicinity of the CDKN2A gene were used to determine whether the mutation in these families is identical by descent, or represents a mutational hotspot in the CDKN2A gene. Our results show that the Val59Gly substitution impairs p16(INK4a) function, and this dysfunction is consistent with structural predictions. All melanoma-affected individuals tested in the families under study harbor this mutation. Interestingly, the Israeli pedigree includes an affected individual who is homozygous for the Val59Gly mutation. A common haplotype of microsatellite markers has been demonstrated for mutation carriers in all four pedigrees. The Israeli pedigree and one of the French melanoma families are of Moroccan and Tunisian Jewish descent, respectively, and the other families originate from regions of France and Spain close to the Pyrenees. We conclude that the Val59Gly mutation is a major contributor to melanoma risk in the families under study and that it may derive from a single ancestral founder of Mediterranean (possibly Jewish) origin.

C7 complement deficiency in an Israeli Arab village.

Deficiencies of terminal complement components, particularly the latter ones, are often detected because of increased susceptibility to Neisserial infections. Herein we document the first report of C7 deficiency among a highly inbred Arab population living in the lower Galilee region of Israel. Both biochemical and molecular analysis were performed on samples from infected survivors and parents of children who succumbed to Neisserial infections in a 4-year period. Only the index case who suffered recurrent infections and a sibling who had not suffered an infection during the outbreak were found to be C7-deficient. The mutation was found to be the one previously described to be prevalent among Israeli Jews of Moroccan ancestry (mutation G1135C). The implications of this finding are discussed in the context of family pedigree, the protective effect of complement deficiency, and the clinical outcome.

[Androgen receptor and male infertility].

The androgen receptor (AR) mediates androgen action determining male sexual phenotypes and promotion of spermatogenesis. Mutations in the AR cause various degrees of androgen resistance resulting in a range of androgen insensitivity syndromes. A single copy gene in the X chromosome encodes the AR. The gene contains a polymorphic triple repeat sequence [(CAG)n] with 9-36 repeats in the normal population, and displays ethnic dependence. In vitro, there is an inverse correlation between CAG repeat length and AR function. Associations exist between short alleles and prostate cancer in men or clinical hyperandrogenism in women. Expansion of the CAG tract > 40 repeats leads to spinal bulbar muscular atrophy (SBMA, Kennedy disease), an adult onset neurodegenerative disease that also presents with low virilization and spermatogenetic defects. The disease may show evidence of anticipation (increasing severity with succeeding generations accompanying further expansion of repeat length). Twelve studies involving Singaporean, Australian, North American and Japanese men reported a relationship between AR CAG repeat length and male infertility, whereas 10 studies, most of them European, found no association. Differences in hereditary or acquired factors in these populations may explain the equivocality. However, statistical methods, sample sizes, study definition and control populations, in addition to laboratory methods vary widely within the published papers, and could affect the results and conclusions. Current data is insufficient to conclude whether IVF patients who display AR CAG expansion may transfer infertility or premutation of neurodegenerative disease to their descendants. We recommend screening of AR CAG repeat length, at least in those populations where an association between repeat length and infertility could be found.

Detection of a long non-coding RNA (CCAT1) in living cells and human adenocarcinoma of colon tissues using FIT-PNA molecular beacons.

Although the function and mechanism of action of long non-coding RNAs (lncRNA) is still not completely known, studies have shown their potential role in the control of gene expression and regulation, in cellular proliferation and invasiveness at the transcriptional level via multiple mechanisms. Recently, colon cancer associated transcript 1 (CCAT1) lncRNA was found to be expressed in colorectal cancer (CRC) tumors but not in normal tissue. This study aimed to study the ability of a CCAT1-specific peptide nucleic acid (PNA) based molecular beacons (TO-PNA-MB) to serve as a diagnostic probe for in vitro, ex vivo, and in situ (human colon biopsies) detection of CRC. The data showed enhanced fluorescence upon in vitro hybridization to RNA extracted from CCAT1 expressing cells (HT-29, SW-480) compared to control cells (SK-Mel-2). Uptake of TO-PNA-MBs into cells was achieved by covalently attaching cell penetrating peptides (CPPs) to the TO-PNA-MB probes. In situ hybridization of selected TO-PNA-MB in human CRC specimens was shown to detect CCAT1 expression in all (4/4) subjects with pre-cancerous adenomas, and in all (8/8) patients with invasive adenocarcinoma (penetrating the bowel wall) tumors. The results showed that CCAT1 TO-PNA-MB is a powerful diagnostic tool for the specific identification of CRC, suggesting that with the aid of an appropriate pharmaceutical vehicle, real time in vivo imaging is feasible. TO-PNA-MB may enable identifying occult metastatic disease during surgery, or differentiating in real time in vivo imaging, between benign and malignant lesions.

The Diagnostic and Prognostic Role of microRNA in Colorectal Cancer - a Comprehensive review.

The discovery of microRNA, a group of regulatory short RNA fragments, has added a new dimension to the diagnosis and management of neoplastic diseases. Differential expression of microRNA in a unique pattern in a wide range of tumor types enables researches to develop a microRNA-based assay for source identification of metastatic disease of unknown origin. This is just one example of many microRNA-based cancer diagnostic and prognostic assays in various phases of clinical research.Since colorectal cancer (CRC) is a phenotypic expression of multiple molecular pathways including chromosomal instability (CIN), micro-satellite instability (MIS) and CpG islands promoter hypermethylation (CIMP), there is no one-unique pattern of microRNA expression expected in this disease and indeed, there are multiple reports published, describing different patterns of microRNA expression in CRC.The scope of this manuscript is to provide a comprehensive review of the scientific literature describing the dysregulation of and the potential role for microRNA in the management of CRC. A Pubmed search was conducted using the following MeSH terms, "microRNA" and "colorectal cancer". Of the 493 publications screened, there were 57 papers describing dysregulation of microRNA in CRC.

Development of a microRNA-based molecular assay for the detection of papillary thyroid carcinoma in aspiration biopsy samples.

BACKGROUND: Although thyroid nodules are common and diagnosed in over 5% of the adult population, only 5% harbor malignancy. Patients with clinically suspicious thyroid nodules need to undergo fine-needle aspiration biopsy (FNAB). The main limitation of FNAB remains indeterminate cytopathology. Only 20%-30% of the indeterminate nodules harbor malignancy, and therefore up to 80% of patients undergo unnecessary thyroidectomy. The aim of this study was to identify and validate a panel of microRNAs (miRNAs) that could serve as a platform for an FNAB-based diagnostic for thyroid neoplasms. METHODS: The study population included 27 consecutive patients undergoing total thyroidectomy for FNAB-based papillary thyroid cancer (n = 20) and benign disorders (n = 7). Aspiration biopsy was performed from the index lesion and from the opposite lobe normal tissue in all study patients at the time of operation. RNA was extracted from all aspiration biopsy samples. Quantitative polymerase chain reaction on a panel of previously selected miRNAs was performed. Polymerase chain reaction results were compared with final histopathology. miRNA from tumor tissues was amplified using the highest value of each miRNA expression in normal tissue as a threshold for malignancy detection. RESULTS: Diagnostic characteristics were most favorable for mir-221 in differentiating benign from malignant thyroid pathology. mir-221 was overexpressed in 19 patients (p < 0.0001) with a sensitive yield of 95%. Specificity, negative and positive predictive value, and accuracy of the miRNA panel were 100%, 96%, 100%, and 98%, respectively. CONCLUSIONS: miRNA quantification for differential diagnosis of thyroid neoplasms within aspiration biopsy samples is feasible and may improve the accuracy of FNAB cytology.

Treatment regimens for Helicobacter pylori infection in children: is in vitro susceptibility testing helpful?

BACKGROUND: Treatment regimens for Helicobacter pylori have variable success rates, and data comparing effectiveness with respect to strain sensitivity are relatively scarce. OBJECTIVE: To evaluate the efficacy of two treatment regimens for eradication of H. pylori and the impact of bacterial susceptibility testing. STUDY DESIGN: 265 children endoscopically diagnosed with H. pylori infection were randomly assigned to receive omeprazole + amoxicillin with clarithromycin or omeprazole + amoxicillin with metronidazole. Bacterial culture and susceptibility was performed in a subgroup. Eradication was assessed by C-urea breath test. RESULTS: Eradication was achieved in 73.4% by omeprazole + amoxicillin with metronidazole and in 62.6% by omeprazole + amoxicillin with clarithromycin (P = 0.078). H. pylori was cultured successfully in 105 patients. Resistance to metronidazole was detected in 31.4% of the isolates and resistance to clarithromycin in 15%. Eradication rate by omeprazole + amoxicillin with metronidazole for metronidazole-susceptible bacteria (N = 38) was 90%, and for resistant bacteria (N = 19) it was 42%. Only 75% of clarithromycin-sensitive strains were successfully treated by omeprazole + amoxicillin with clarithromycin, and none of the cases with clarithromycin-resistant strains responded to omeprazole + amoxicillin with clarithromycin treatment. CONCLUSION: There is a trend of greater efficacy of eradication with omeprazole + amoxicillin with metronidazole versus omeprazole + amoxicillin with clarithromycin therapy. Although resistance negatively influences eradication, first-line sensitivity-based treatment would be expected to improve this rate only slightly. Susceptibility testing should probably be reserved only for treatment failures.

Helicobacter pylori genotypes in Israeli children: the significance of geography.

BACKGROUND: There is substantial genetic variation among different isolates of Helicobacter pylori, which may affect the clinical outcome. The aims of this study were to find the common H. pylori genotypes in Israeli children and to look for a possible genotype-phenotype correlation. METHODS: Ninety-eight H. pylori cultures were isolated from antral biopsy specimens of symptomatic Israeli children and were analyzed for vacA and iceA genotype and cagA and cagE status by polymerase chain reaction. RESULTS: cagA and cagE genes were present in only 25.5% and 24.5%, respectively. The common vacA genotype was s2m2, which was found in 65%. Eleven specimens (11%) contained multiple vacA genotypes. iceA1 was found in 37% and iceA2 in 52% of cases. Both iceA alleles were found in 11%. Increased prevalence of iceA1 and cagE were observed in children with duodenal disease, although it did not reach significance. CONCLUSIONS: The low prevalence of cagA and the high prevalence of vacA genotype s2m2 in Israeli pediatric patients are different from the genotype prevalence reported globally. However, similar findings have been reported in Egypt, indicating a possible geographic influence. There is a possible correlation between duodenal ulcer and cag E and ice A1 genotype, but the power of the study was too low to prove it.