The non-vesicle cell-free DNA (cfDNA) induces cell transformation associated with horizontal DNA transfer.

BACKGROUND: Cell-free DNA (cfDNA) is a source for liquid biopsy used for cancer diagnosis, therapy selection, and disease monitoring due to its non-invasive nature and ease of extraction. However, cfDNA also participates in cancer development and progression by horizontal transfer. In humans, cfDNA circulates complexed with extracellular vesicles (EV) and macromolecular complexes such as nucleosomes, lipids, and serum proteins. The present study aimed to demonstrate whether cfDNA not associated with EV induces cell transformation and tumorigenesis. METHODS: Supernatant of the SW480 human colon cancer cell line was processed by ultracentrifugation to obtain a soluble fraction (SF) and a fraction associated with EV (EVF). Primary murine embryonic fibroblast cells (NIH3T3) underwent passive transfection with these fractions, and cell proliferation, cell cycle, apoptosis, cell transformation, and tumorigenic assays were performed. Next, cfDNA was analyzed by electronic microscopy, and horizontal transfer was assessed by human mutant KRAS in recipient cells via PCR and recipient cell internalization via fluorescence microscopy. RESULTS: The results showed that the SF but not the EVF of cfDNA induced proliferative and antiapoptotic effects, cell transformation, and tumorigenesis in nude mice, which were reduced by digestion with DNAse I and proteinase K. These effects were associated with horizontal DNA transfer and cfDNA internalization into recipient cells. CONCLUSIONS: The results suggest pro-tumorigenic effects of cfDNA in the SF that can be offset by enzyme treatment. Further exploration of the horizontal tumor progression phenomenon mediated by cfDNA is needed to determine whether its manipulation may play a role in cancer therapy.

Knowledge, attitudes, and behavior regarding antibiotics, antibiotic use, and antibiotic resistance in students and health care professionals of the district of Barranquilla (Colombia): A cross-sectional survey.

Objective: To evaluate the knowledge, attitudes and behavior regarding antibiotics, use of antibiotics, and antibiotic resistance in students and health care professionals of the district of Barranquilla, Colombia. Study design: Descriptive, cross-sectional. Methods: A sample of 399 respondents was selected, that included health professionals and medical students from 12 health institutions in the district of Barranquilla (Colombia), using an established stratified sampling method. Each of the respondent professionals completed a survey that included 43 items in the Likert scale. A descriptive analysis of the study variables was performed using the software SPSS version 25. Results: Most of the respondents were women (64.4%), aged between 26 and 35 years (47.6%); 28.8% were nurses and 26.1% general practitioners, with ≤10 years of professional experience (63.4%). Overall, the survey revealed that the participants had considerable knowledge about antibiotic use (89.5%-98% correct answers) and the spread of antibiotic resistance (67.4%-89% correct answers). Approximately 74% of the respondents agreed or fully agreed with the questions related to the management of infections and the provision of advice. Conclusions: The present study revealed that most of the health care professionals surveyed had a good knowledge about antibiotic use, although strategies must be developed to strengthen knowledge regarding the spread of antibiotic resistance. Likewise, it is important to identify opportunities for improvement related with access to the guidelines and/or materials necessary to treat infections and to provide advice on antibiotic use and antibiotic resistance.

Hereditary diffuse gastric cancer (HDGC). An overview.

It is estimated that up to 10% of gastric carcinomas show familial aggregation. In contrast, around 1-3 % (approximately 33,000 yearly) are genuinely hereditary. Hereditary diffuse gastric cancer (HDGC) is a rare malignancy characterized by autosomal dominant inheritance of pathological variants of the CDH1 and CTNNA1 genes encoding the adhesion molecules E-cadherin and α-catenin, respectively. The multifocal nature of the disease and the difficulty of visualizing precursor lesions by endoscopy underscore the need to be aware of this malignancy as surgical prevention can be fully protective. Here, we provide an overview of the main epidemiological, clinical, genetic, and pathological features of HDGC, as well as updated guidelines for its diagnosis, genetic testing, counseling, surveillance, and management. We conclude that HDGC is a rare, highly penetrant disease that is difficult to diagnose and manage, so it is necessary to correctly identify it to offer patients and their families' adequate management following the recommendations of the IGCL. A critical point is identifying a mutation in HDGC families to determine whether unaffected relatives are at risk for cancer.

Selection and characterization of highly specific recombinant antibodies against West Nile Virus E protein.

West Nile virus is a widespread mosquito-borne human and animal pathogenic virus of increasing importance. The E protein of the viral envelope is critical for attachment and entry into the host cell and has been the target for vaccine design and small molecule inhibitors. Furthermore, the detection of anti-E IgM and IgG antibodies is widely used in serology to diagnose these infections. Here we describe a strategy for the production of recombinant antibodies against the E protein of West Nile virus for research and immunodiagnostic purposes. Initially the fast and easy protocol previously developed for the similar Tick-borne encephalitis virus has been adapted to West Nile virus E antigen production and purification. A human naïve scFv phage library has been selected on the produced antigen, identifying a panel of highly specific anti-E protein antibodies. Once produced as scFv-Fc recombinant proteins, the selected antibodies have been characterized by mapping their binding sites and by defining their affinity for the target. The impact on neutralizing virus attachment and entry has been also evaluated. The obtained results demonstrate the potential of the produced reagents for research and diagnostic applications.

A method for rapid and high-yield production of the tick-borne encephalitis virus E and DIII recombinant proteins in E. coli with preservation of the antigenic properties.

Tick-borne encephalitis virus (TBEV) is a member of the Flavivirus genus and is the main pathogenic arbovirus circulating in Europe, Russia and China. The envelope (E) protein is exposed on the viral surface and is the main antigen that is employed in diagnostic tests based on the detection of protein-specific antibodies from serum samples of infected individuals. The high degree of similarity among the E proteins of flaviviruses can, in some cases, lead to cross-reactivity and false-positive results in serological tests. Increased specificity in the detection of positive sera for different Flavivirus infections is often obtained by using a portion of the E protein, namely, the DIII domain. Different strategies and expression systems have been described for E and DIII protein production. Here, we present the optimization of an easy and fast method for TBEV E and DIII antigen production and partial purification from E. coli inclusion bodies. The antigenic properties of the produced antigens are retained, as validated by ELISAs with anti-TBEV murine sera as well as sera from infected human patients. The potential applications of both proteins as diagnostic reagents were confirmed.

Emisiones antrooogénicas de amoniaco, nitratos óxido nitroso: compuestos nitrrogenados que afectan el medio ambiente en el sector agropecuario Colmbbiano / Anthrpogenic emiissions of ammonia, nitrates and nitrous oxide: nitrogen compounds that affect the environment in the Colombian agricultural sector

El cambio climático es un fenómeno global actual que posee diversas causas, tales como el incremento antropogénico en las emisiones de gases de efecto invernadero y su influencia en el planeta. El óxido nitroso (N2O) es un gas con un potencial de calentamiento 298 veces mayor que el CO2; es emitido por diversos sectores económicos, entre ellos la agricultura y la ganadería por el empleo de fertilizantes nitrogenados. Este uso no incrementa únicamente las emisiones de N2O, sino que afecta todos los eslabones del ciclo del nitrógeno, siendo importante reconocer las interacciones entre sus reacciones (amonificación, nitrificación, denitrificación, etc.) para comprender los efectos sobre la lixiviación de nitratos, volatilización de amoniaco y emisiones de N2O, tres compuestos naturales que afectan el medio ambiente. En Colombia, el IDEAM reportó que en 2004 el sector agropecuario nacional produjo 94,91 gigagramos (Gg) de N2O, pues los sistemas de producción agrícolas y bovinos utilizan altas cantidades de fertilizantes nitrogenados con bajos índices de asimilación. Por ello adquiere importancia implementar buenas prácticas que midan y mitiguen estas emisiones en campo, buscando mayor eficiencia en la transformación de componentes nitrogenados dentro del sistema. En este artículo se enfatiza un manejo adecuado del suelo y la utilización de inhibidores de nitrificación naturales (presentes en algunas plantas tropicales) o químicos, como la nitrapyrina, DCD y NBPT (este último como inhibidor de la amonificación), observando su forma de acción y medición, su efecto mitigador y los incrementos productivos. La presente es una revisión bibliográfica que aborda la problemática ambiental de la ganadería bovina.

Climate change is a global effect currently present with several causes, such as the increased of anthropogenic emissions of greenhouse gases and its influence on the planet. Nitrous oxide (N2O) is a gas with a global warming potential 298 times greater than CO, which comes of many sectors, one of them is the agriculture and livestock through the use of nitrogen fertilizers. This use not only increases N2O emissions, its affects all stages of the nitrogen cycle, being important to observe the interactions between their reactions (ammonification, nitrification, denitrification etc.) to understand the effects over nitrate leaching, ammonia volatilization and N2O emissions, three natural chemicals that affect the environment. In Colombia, IDEAM reports that for 2004, the national agricultural sector produced 94.91 Gg of N2O, knowing the agricultural and livestock productions use high quantities of nitrogen fertilizers, with low assimila2tion rates. Therefore, some adaptable practices and methodologies become important to measure and mitigate these emissions in the field, seeking greater efficiency in the transformation of nitrogen components in the system, being reviewed on this paper the appropriate land management or implementation of natural nitrification inhibitors (present in some tropical plants) or chemical inhibitors, such as nitrapyrina, DCD and NBPT (as an ammonification inhibitor). This is a literature review that approaches the environmental problems of livestock.

Role of TNF in sickness behavior and allodynia during the acute phase of Chagas' disease.

Chagas disease, caused by the intracellular protozoan Trypanosoma cruzi, is associated with inflammation, discomfort and pain during the acute phase. The influence of TNF-α (tumor necrosis factor) in this disease outcome is controversial. In this way, the aim of this work was to determine the role of the TNF-α blocker etanercept in the pain, discomfort, and survival during the Chagas' acute phase of mice experimentally infected with a wild virulent strain of T. cruzi. The infection with this wild strain was responsible for a severe visceral inflammation and said parasite showed a tropism in peritoneal fluid cells. Etanercept was able to restore spontaneous vertical and horizontal activities during the second week after infection and to abolish mechanical allodynia during the first week after infection. Finally, etanercept delayed the mortality without any effect on the parasitemia rates. This is the first report that correlates sickness behavior and allodynia with TNF-α and suggests that this cytokine may play an important role in the physiopathology of the acute phase.

CALIDAD DE LA LECHE, DIGESTIBILIDAD IN VITRO DE LA MATERIA SECA Y PRODUCCIÓN EN VACAS ALIMENTADAS CON GRAMÍNEAS SOLAS O ASOCIADAS CON Lotus uliginosus / MILK CHEMICAL COMPOSITION, MATTER DRY IN VITRO DIGESTIBILITY AND PRODUCTION IN COWS FED ALONE GRASSES OR ASSOCIATED Lotus uliginosus

La presente investigación tuvo como objetivo evaluar el uso de la leguminosa Lotus uliginosus como alternativa de inclusión dentro de las praderas en dos ecorregiones estratégicas de la Sabana de Bogotá. En cada finca se evaluaron cuatro tratamientos: 1) kikuyo solo (P. clandestinum), 2) kikuyo asociado con trébol pata de pájaro (P. clandestinum + L. uliginosus), 3) festuca alta sola (F. arundinacea) y 4) Festuca alta asociada con trébol pata de pájaro (F. arundinacea + L. uliginosus). El número de bovinos varió de acuerdo a la capacidad de carga de cada finca. Se utilizaron animales que se encontraban en el segundo tercio de lactancia durante un período experimental de siete días. En cada tratamiento se determinó la producción (L/día) y calidad de leche (% de grasa, proteína y sólidos totales); en la pastura se evaluó la composición química (MS, FDN, FDA) y la digestibilidad in vitro de la materia seca (DIVMS). Se midió el consumo voluntario. En la finca Megaleche la producción de biomasa presentó diferencias significativas (P<0,05) entre los tratamientos destacándose el tratamiento del kikuyo solo; la calidad nutricional de las pasturas también presentó diferencias (P<0,04) destacándose el tratamiento de kikuyo + trébol pata de pájaro, asociación que también se destacó en consumo voluntario (P< 0.05) y producción de leche (P<0,05). En la finca Colega el contenido de proteína del forraje presentó diferencias cuando las gramíneas se asociaron con el trébol pata de pájaro (P<0,001). Se concluyó que la introducción del Lotus en las praderas mejoró la producción de leche y consumo de materia seca en los animales.

The present research aimed to evaluate the use of the legume Lotus uliginosus alternatively inclusion into the meadows on two strategic ecoregions savannah of Bogota. In each farm four treatments: 1) only kikuyu (P. clandestinum), 2) associated kikuyo Birdsfoot trefoil (P. clandestinum + L. uliginosus), 3) single tall fescue (F. arundinacea) and 4) tall fescue associated Birdsfoot trefoil (F. arundinacea + L. uliginosus). The number of cattle varied according the load capacity of each farm. Were used animals that were in the second third of lactation during a trial period of seven days, in each treatment was determined production (L/day) and milk quality (% of fat, protein and total solids). In the pasture it was evaluated chemical composition (DM, NDF, ADF) and in vitro digestibility of matter dry (IVDMD). Voluntary intake was measured. On the farm Megaleche biomass production showed significant differences (P˂0.05) between treatments kikuyu emphasizing treatment alone nutritional quality of pastures also showed significant differences (P˂0.04) emphasizing treating kikuyu + Birdsfoot trefoil an association that was also highlighted in the voluntary intake (P˂0.05) and milk production (P˂0.05). On the farm Colega protein content of forage grasses differed when associated with Birdsfoot trefoil (P˂0.001). It was concluded that the introduction of Lotus grassland improved milk production and dry matter intake in animals.

Upregulation of NKG2D ligands and enhanced natural killer cell cytotoxicity by hydralazine and valproate.

Natural killer cells play a role in the immune antitumor response by recognizing and eliminating tumor cells through the engagement of NKG2D receptors with their ligands on target cells. This work aimed to investigate whether epigenetic drugs are able to increase MICA and MICB expression as well as NK cell cytotoxicity. Prostate, colon, breast and cervical cancer cell lines were analyzed for the expression of MICA and MICB at the mRNA and protein levels by RT-PCR, Western blot, flow cytometry and ELISA. The activating mark H3K4m2 at the MICA and MICB promoters was investigated by ChIP assays. Cytotoxicity of NK cells against the target epithelial cancer cells was investigated with the CD107 cytotoxicity assay. The results show that hydralazine and valproic acid not only increase the expression of MICA and MICB ligands of target cells, but also reduce their shedding to the supernatant. This upregulation occurs at the transcriptional level as revealed by increase of the H3K4 activating mark at the promoter of MICA and MICB genes. These effects are paralleled by increased cytotoxicity of NK cells, which was attenuated at different degrees by using blocking antibodies against the NKG2D receptor and ligands. In conclusion, our results demonstrate the ability of hydralazine and valproate to increase the NK activity against epithelial cancer cell lines and suggest that these drugs could reduce the levels of soluble MICA and MICB helping in avoiding tumor-induced suppression of NK cytotoxicity against the tumor.

Pharmacokinetics of hydralazine, an antihypertensive and DNA-demethylating agent, using controlled-release formulations designed for use in dosing schedules based on the acetylator phenotype.

PURPOSE: The antihypertensive hydralazine has recently been repositioned as DNA demethylating for the epigenetic therapy of cancer. As the acetylator phenotype is the key determinant of its plasma levels, the dose of hydralazine needs to be adjusted for the acetylation status of patients. METHODS: The pharmacokinetics of orally administered hydralazine was evaluated in 26 healthy volunteers (13 slow and 13 fast acetylators) after a single dose of 182 mg administered as a controlled-release tablet. Plasma levels of hydralazine were analyzed in 85 cancer patients treated with this formulation at a dose of 83 mg/day and 182 mg/day for slow and fast acetylators, respectively. RESULTS: The C(max) and t(max) of hydralazine for fast acetylators were 208.4 ± 56.9 SD ng/ml and 2.8 ± 2.5 h, respectively. The corresponding results for slow acetylators were 470.4 ± 162.8 ng/ml, and 4.4 ± 3.1 h. Healthy volunteers who were fast acetylators had no clinically significant changes in blood pressure and heart rate or any other side-effect, however, slow acetylators had transient episodes of headache, tachycardia and faintness. Among 85 cancer patients that received either 182 mg or 83 mg of hydralazine daily, according to their acetylator status, the mean concentrations of hydralazine in plasma were 239.1 ng/ml and 259.2 ng/ml for fast and slow acetylators, respectively. These differences were not significantly different, p = 0.3868. CONCLUSIONS: The administration of dose-adjusted controlled-release hydralazine according to the acetylation status of cancer patients yields similar levels of hydralazine.

[Endoscopic decompression of the ulnar nerve in the cubital tunnel syndrome: about 55 patients]. / Décompression endoscopique du nerf ulnaire dans le syndrome cubital au coude: à propos de 55 patients.

INTRODUCTION: Sulcus ulnaris syndrome is the second most common neurocompression syndrome in the upper limb after carpal tunnel syndrome. Its severity can be appreciated by the Dellon Classification. We present our experience and results after endoscopic decompression. PATIENTS AND METHODS: A retrospective chart review of 55 patients operated over a 3-year period was performed. The patients, 37 men and 18 women, had an average age of 54 years (range: 27-82 years) at the time of surgery. The clinical diagnostic was always confirmed by a neurophysiological examination of the nerve conduction. According to the Dellon Classification, 11 patients had mild sulcus ulnaris syndrome, 31 had moderate and 13 had severe. The mean follow-up time was 21 months (range: 6-42 months). RESULTS: The sensibility was normalised in 85% of the patients. Compared to the contralateral non-operated side, the mean grip strength improved from 68 to 94% and the mean pinch grip from 72 to 95%. The rate of nerve luxation did not change (5.5%). According to the modified Bishop rating system, 38 patients (69%) had excellent, 13 patients (23.5%) good and four patients (7.5%) fair results. One haematoma necessitating a revision, a minimal lesion of the ulnar nerve with restitution ad integrum and a hypoesthesia of the elbow, occurred after surgery resulting in a complication rate of 5.5%. Ninety-eight percent of the patients responded that they would undergo the endoscopic procedure again if needed. CONCLUSION: Endoscopic decompression of the ulnar nerve in sulcus ulnaris syndrome is very well appreciated by patients and also provides promising clinical results.

Infrared photoluminescence of composite films containing quasi-isolated multiwalled carbon nanotubes and carbon nanoshells.

Films of small irregular-shaped aggregates composed by several layers of turbostratic graphite encapsulating iron nanoparticles, like carbon nanoshells, and quasi-isolated multi-walled carbon nanotubes (MWCNTs), were synthesized by the chemical vapor deposition method on glass substrates. Quasi-isolated MWCNTs were found emerging in different directions on the agglomerates composed of carbon nanoshells. The morphological properties of the films were characterized using scanning electron microscopy, high-resolution transmission electron microscopy and Raman spectroscopy, whereas their optical properties were investigated using optical absorption and photoluminescence (PL) spectroscopy. The studies show a high-intensity PL signal in the infrared at room temperature. This PL signal shows several peaks, which cannot be explained by a blackbody emission. However, the overall PL signal could be a combination of the black emission and electronic transitions. Furthermore, the observed infrared PL emission could be ideal for potential applications such as in gas sensors, infrared detectors and so on.

Epigenetic therapy and cisplatin chemoradiation in FIGO stage IIIB cervical cancer.

INTRODUCTION: This trial aimed to evaluate the safety and efficacy of epigenetic therapy associated with cisplatin chemoradiation in FIGO Stage IIIB patients. METHODS: Hydralazine containing either 182 mg for rapid-, or 83 mg for slow acetylators and magnesium valproate were administered at 30 mg/kg tid. Both drugs were taken until intracavitary therapy was finished. Pelvic external beam radiation and low-dose rate brachytherapy were administered at a total cumulative dose to point A of at least 85 Gy. Weekly cisplatin at 40 mg/m2 was delivered for six cycles. RESULTS: Twenty-two patients were included and 18 (82%) patients completed treatment. Mean dose to point A was 84.6 + 2.2. Median number of cisplatin cycles was 5.5 (range, 1-6). Brachytherapy was delayed for technical reasons; the mean overall treatment time was 11.8 weeks. Grade 3 anemia, leucopenia, neutropenia, and thrombocytopenia were observed in 9%, 45%, 45%, and 9% of patients, respectively. CONCLUSIONS: Hydralazine and valproate are well-tolerated and safe when administered with cisplatin chemoradiation. Unfortunately, the suboptimal administration of brachytherapy for technical reasons in this study, precluded assessing the efficacy of epigenetic therapy. However, the tolerability of this regimen administered concurrent to radiation needs to be further tested.

[Assessment of patients with distal radius fracture treated with percutaneous fixation]. / Evaluación de pacientes con fractura de radio distal tratados con fijación percutánea.

UNLABELLED: The trend in distal radius fracture is towards open reduction and internal fixation; however, treatment with closed reduction and percutaneous nailing continues to provide good results, but physical therapy and appropriate early rehabilitation are needed. MATERIAL AND METHODS: We assessed patients with distal radius fractures treated with reduction with percutaneous maneuvers and nailing; we used clinical functional scales, like the DASH, the Mayo Clinic wrist scale, and the Visual Analog Scale (VAS) to assess pain. Patient results were compared at 12 and 24 weeks. We also compared patients with and without rehabilitation. RESULTS: A total of 60 patients were assessed using the DASH scale; patients with rehabilitation had a score of 4.3, while those without rehabilitation had a score of 10.5 (p = 0.00001). In the Mayo Clinic scale, patients with rehabilitation had a score of 86.7, while those without rehabilitation had a score of 77.8 (p = 0.00001). The VAS was not significant. CONCLUSION: All patients showed improvement in their clinical assessment scales. However, when patients with and without rehabilitation are compared, the difference was greater in the Mayo Clinic scale and in the DASH scale.

Pharmacogenetics and pharmacoepigenetics of gemcitabine.

Gemcitabine (2',2'-difluoro 2'deoxycytidine, dFdC) is an analog of cytosine with distinctive pharmacological properties and a wide antitumor-activity spectrum. The pharmacological characteristics of gemcitabine are unique because two main classes of genes are essential for its antitumor effects: membrane transporter protein-coding genes, whose products are responsible for drug intracellular uptake, as well as enzyme-coding genes, which catalyze its activation and inactivation. The study of the pharmacogenetics and pharmacoepigenetics of these two gene classes is greatly required to optimize the drug's therapeutic use in cancer. This review aims to provide an update of genetic and epigenetic bases that may account for interindividual variation in therapeutic outcome exhibited by gemcitabine.