Amoxicillin and Clarithromycin Mucoadhesive Delivery System for Helicobacter pylori Infection in a Mouse Model: Characterization, Pharmacokinetics, and Efficacy.

Helicobacter pylori is the main pathogen responsible for gastric ulcers and a predisposing factor of stomach cancer. Although current treatment is usually successful, it requires high doses and frequent administration. An innovative mucoadhesive system (Mucolast®) loaded with amoxicillin and clarithromycin is proposed to improve the efficacy of treatment against H. pylori. The drug product was optimized based on its viscoelastic properties to obtain long-term stability of the vehicle. The drug release mechanisms were different for both antibiotics based on their solubilization status. A systemic and stomach pharmacokinetic profile was obtained after three different doses were administered to mice, obtaining similar systemic exposure levels but an increase in drug concentration in the stomach. The efficacy results in mice infected with H. pylori also demonstrated the superiority of the antibiotics when administered in Mucolast®, as shown by the bacterial count in stomach tissue and under histopathological and biochemical evaluation. The proposed treatment was efficacious and safe and is presented as a realistic alternative to current treatment options to improve patient compliance and to reduce bacterial resistance.

Oncogénesis del pólipo endometrial. Manejo clínico a través de marcadores biomoleculares / Oncogenesis of the endometrial polyp. Clinical management through biomolecular markers

La prevalencia de los pólipos endometriales se estima entre el 7,8 y el 35% de las mujeres, siendo mayor en el estado posmenopáusico. Los pólipos endometriales se asocian con hiperplasia endometrial y carcinogénesis, con una prevalencia informada de lesiones malignas y premalignas que puede llegar al 13%. La detección de pólipos endometriales en edad peri o posmenopáusica, en pacientes sintomáticas o asintomáticas, requiere un examen histeroscópico meticuloso. No está claro si la polipectomía debe realizarse de forma rutinaria en pacientes asintomáticas. El manejo expectante de pólipos pequeños y asintomáticos es razonable en muchos casos. Se necesitan estudios adicionales para dilucidar si los pólipos endometriales son precursores de cáncer, o simplemente marcadores de una enfermedad endometrial. Los biomarcadores capaces de detectar cambios a nivel molecular en los pólipos y el tejido endometrial nos ayudan a un mejor conocimiento y clasificación de los procesos malignos. Este conocimiento permite pasar de una medicina intervencionista a una medicina más conservadora, basada en la confianza de un conocimiento más preciso de los procesos biomoleculares. GynEC®-DX se basa en determinar la expresión de genes que se combinan en un algoritmo matemático diagnóstico para llegar a un diagnóstico negativo o positivo de cáncer de endometrio. La prueba molecular tiene un valor predictivo negativo del 99,6%, con una alta especificidad y sensibilidad. Esta prueba podría usarse para el diagnóstico diferencial del cáncer de endometrio en mujeres con pólipos endometriales sin requerir la exéresis de pólipos, limitando el riesgo iatrogénico y evitando intervenciones innecesarias

The prevalence of endometrial polyps is estimated between 7.8 and 35% of women, being more prevalent in postmenopausal women. Endometrial polyps are associated with endometrial hyperplasia and carcinogenesis, with an informed prevalence of malignant and premalignant lesions that may reach 13%. The detection of endometrial polyps in peri- or postmenopausal (status) age, in symptomatic or asymptomatic patients, requires a meticulous hysteroscopic examination. It is unclear if routine polypectomy should be performed in asymptomatic patients. The expectant management of small and asymptomatic polyps is reasonable in many cases. Additional studies are needed to elucidate whether endometrial polyps are precursors of cancer, or simply markers of an endometrial disease. Biomarkers capable of detecting changes at the molecular level in polyps and endometrial tissue help us to better understand and classify malignant processes. This knowledge allows to move from an interventional medicine to a more conservative medicine, based on the confidence of a more precise knowledge of the biomolecular processes. GynEC®-DX is based on determining the expression of genes that are combined in a diagnostic mathematical algorithm to arrive at a negative or positive diagnosis of endometrial cancer. The molecular test has a negative predictive value of 99.6%, with high specificity and sensitivity. This test could be used for the differential diagnosis of endometrial cancer in women with EPs and prevent the resection of polyps, limiting the iatrogenic risk and avoiding unnecessary interventions

Detection of collagen triple helix repeat containing-1 and nuclear factor (erythroid-derived 2)-like 3 in colorectal cancer.

BACKGROUND: Collagen Triple Helix Repeat Containing-1 (CTHRC1) and Nuclear factor (erythroid-derived 2)-like 3 (NFE2L3) may be useful biomarker candidates for the diagnosis of colorectal cancer (CRC) since they have shown an increase messenger RNA transcripts (mRNA) expression level in adenomas and colorectal tumours when compared to normal tissues. METHODS: To evaluate CTHRC1 and NFE2L3 as cancer biomarkers, it was generated and characterised several novel specific polyclonal antibodies (PAb), monoclonal antibodies (MAbs) and soluble Fab fragments (sFabs) against recombinant CTHRC1 and NFE2L3 proteins, which were obtained from different sources, including a human antibody library and immunised animals. The antibodies and Fab fragments were tested for recognition of native CTHRC1 and NFE2L3 proteins by immunoblotting analysis and enzyme-linked immunosorbent assay (ELISA) in colorectal cell lines derived from tumour and cancer tissues. RESULTS: Both, antibodies and a Fab fragment showed high specificity since they recognised only their corresponding recombinant antigens, but not a panel of different unrelated- and related proteins.In Western blot analysis of CTHRC1, a monoclonal antibody designated CH21D7 was able to detect a band of the apparent molecular weight of a full-length CTHRC1 in the human colon adenocarcinoma cell line HT29. This result was confirmed by a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with the monoclonal antibodies CH21D7 and CH24G2, detecting CTHRC1 in HT29 and in the colon adenocarcinoma cell line SW620.Similar experiments were performed with PAb, MAbs, and sFab against NFE2L3. The immunoblot analysis showed that the monoclonal antibody 41HF8 recognised NFE2L3 in HT29, and leukocytes. These results were verified by DAS-ELISA assay using the pairs PAb/sFab E5 and MAb 41HF8/sFab E5.Furthermore, an immunoassay for simultaneous detection of the two cancer biomarkers was developed using a Dissociation-Enhanced Lanthanide Fluorescent Immunoassay technology (DELFIA). CONCLUSIONS: In conclusion, the antibodies obtained in this study are specific for CTHRC1 and NFE2L3 since they do not cross-react with unrelated- and related proteins and are useful for specific measurement of native CTHRC1 and NFE2L3 proteins. The antibodies and immunoassays may be useful for the analysis of CTHRC1 and NFE2L3 in clinical samples and for screening of therapeutic compounds in CRC.

Molecular markers of endometrial carcinoma detected in uterine aspirates.

Endometrial cancer (EC) is the most frequent of the invasive tumors of the female genital tract. Although usually detected in its initial stages, a 20% of the patients present with advanced disease. To date, no characterized molecular marker has been validated for the diagnosis of EC. In addition, new methods for prognosis and classification of EC are needed to combat this deadly disease. We thus aimed to identify new molecular markers of EC and to evaluate their validity on endometrial aspirates. Gene expression screening on 52 carcinoma samples and series of real-time quantitative PCR validation on 19 paired carcinomas and normal tissue samples and on 50 carcinoma and noncarcinoma uterine aspirates were performed to identify and validate potential biomarkers of EC. Candidate markers were further confirmed at the protein level by immunohistochemistry and Western blot. We identified ACAA1, AP1M2, CGN, DDR1, EPS8L2, FASTKD1, GMIP, IKBKE, P2RX4, P4HB, PHKG2, PPFIBP2, PPP1R16A, RASSF7, RNF183, SIRT6, TJP3, EFEMP2, SOCS2 and DCN as differentially expressed in ECs. Furthermore, the differential expression of these biomarkers in primary endometrial tumors is correlated to their expression level in corresponding uterine fluid samples. Finally, these biomarkers significantly identified EC with area under the receiver-operating-characteristic values ranging from 0.74 to 0.95 in uterine aspirates. Interestingly, analogous values were found among initial stages. We present the discovery of molecular biomarkers of EC and describe their utility in uterine aspirates. These findings represent the basis for the development of a highly sensitive and specific minimally invasive method for screening ECs.

Genomic resources for a commercial flatfish, the Senegalese sole (Solea senegalensis): EST sequencing, oligo microarray design, and development of the Soleamold bioinformatic platform.

BACKGROUND: The Senegalese sole, Solea senegalensis, is a highly prized flatfish of growing commercial interest for aquaculture in Southern Europe. However, despite the industrial production of Senegalese sole being hampered primarily by lack of information on the physiological mechanisms involved in reproduction, growth and immunity, very limited genomic information is available on this species. RESULTS: Sequencing of a S. senegalensis multi-tissue normalized cDNA library, from adult tissues (brain, stomach, intestine, liver, ovary, and testis), larval stages (pre-metamorphosis, metamorphosis), juvenile stages (post-metamorphosis, abnormal fish), and undifferentiated gonads, generated 10,185 expressed sequence tags (ESTs). Clones were sequenced from the 3'-end to identify isoform specific sequences. Assembly of the entire EST collection into contigs gave 5,208 unique sequences of which 1,769 (34%) had matches in GenBank, thus showing a low level of redundancy. The sequence of the 5,208 unigenes was used to design and validate an oligonucleotide microarray representing 5,087 unique Senegalese sole transcripts. Finally, a novel interactive bioinformatic platform, Soleamold, was developed for the Senegalese sole EST collection as well as microarray and ISH data. CONCLUSION: New genomic resources have been developed for S. senegalensis, an economically important fish in aquaculture, which include a collection of expressed genes, an oligonucleotide microarray, and a publicly available bioinformatic platform that can be used to study gene expression in this species. These resources will help elucidate transcriptional regulation in wild and captive Senegalese sole for optimization of its production under intensive culture conditions.