Molecular docking analysis of chlorpyrifos at the human α7-nAChR and its potential relationship with neurocytoxicity in SH-SY5Y cells.

The organophosphate insecticide chlorpyrifos (CPF), an acetylcholinesterase inhibitor, has raised serious concerns about human safety. Apart from inducing synaptic acetylcholine accumulation, CPF could also act at nicotinic acetylcholine receptors, like the α7-isoform (α7-nAChR), which could potentially be harmful to developing brains. Our aims were to use molecular docking to assess the binding interactions between CPF and α7-nAChR through, to test the neurocytotoxic and oxidative effects of very low concentrations of CPF on SH-SY5Y cells, and to hypothesize about the potential mediation of α7-nAChR. Docking analysis showed a significant binding affinity of CPH for the E fragment of the α7-nAChR (ΔGibbs: -5.63 to -6.85 Kcal/mol). According to the MTT- and Trypan Blue-based viability assays, commercial CPF showed concentration- and time-dependent neurotoxic effects at a concentration range (2.5-20 µM), ten-folds lower than those reported to have crucial effects for sheer CPF. A rise of the production of radical oxygen species (ROS) was seen at even lower concentrations (1-2.5 µM) of CPF after 24h. Notably, our docking analysis supports the antagonistic actions of CPF on α7-nAChR that were recently published. In conclusion, while α7-nAChR is responsible for neuronal survival and neurodevelopmental processes, its activity may also mediate the neurotoxicity of CPF.

Molecular Docking Analysis at the Human α7-nAChR and Proliferative and Evoked-Calcium Changes in SH-SY5Y Cells by Imidacloprid and Acetamiprid Insecticides.

Acetamiprid (ACE) and Imidacloprid (IMI) are widely-used neonicotinoid insecticides (NNIs) with functional activity at human acetylcholine nicotinic receptors and, therefore, with putative toxic effects. The objective of this study was the evaluation of the interactions between NNIs and α7-nAChR, as this receptor keeps intracellular Ca2+ ([Ca2+]i) to an optimum for an adequate neuronal functioning. Possible interactions between NNIs and the cryo-EM structure of the human α-7 nAChR were identified by molecular docking. Additionally, NNI effects were analyzed in neuroblastoma SH-SY5Y cells, as they naturally express α-7 nAChRs. Functional studies included proliferative/cytotoxic effects (MTT test) in undifferentiated SH-SY-5Y cells and indirect measurements of [Ca2+]i transients in retinoic acid-differentiated SH-SY-5Y cells loaded with Fluo-4 AM. Docking analysis showed that the binding of IMI and ACE occurred at the same aromatic cage that the specific α-7 nAChR agonist EVP-6124. IMI showed a better docking strength than ACE. According to the MTT assays, low doses (10-50 µM) of IMI better than ACE stimulated neuroblastoma cell proliferation. At higher doses (250-500 µM), IMI also prevailed over ACE and dose-dependently triggered more abrupt fluorescence changes due to [Ca2+]i mobilization in differentiated SH-SY5Y neurons. Indeed, only IMI blunted nicotine-evoked intracellular fluorescence stimulation (i.e., nicotine cross-desensitization). Summarizing, IMI demonstrated a superior docking strength and more robust cellular responses compared to ACE, which were likely associated with a stronger activity at α-7nAChRs. Through the interaction with α-7nAChRs, IMI would demonstrate its high neurotoxic potential for humans. More research is needed for investigating the proliferative effects of IMI in neuroblastoma cells.

Neurocytotoxicity of imidacloprid- and acetamiprid-based comercial insecticides over the differentiation of SH-SY5Y neuroblastoma cells.

Neonicotinoids are effective insecticides with specificity for invertebrate nicotinic acetylcholine receptors. Neonicotinoids are chemically stable and tend to remain in the environment for long so concerns about their neurotoxicity in humans do nothing but increase. Herein, we evaluated the chronic toxic effects of acetamiprid- and imidacloprid-based insecticides over the differentiation of human neuroblastoma SH-SY5Y cells, which were exposed to these insecticides at a concentration range similar to that applied to crop fields (0.01-0.5 mM). Both insecticides did not have acute cytotoxic effects in both non-differentiated and in staurosporine-differentiated SH-SY5Y cells cytotoxicity as measured by the MTT and vital-dye exclusion tests. However, after a chronic (7-day) treatment, only imidacloprid dose-dependently decreased the viability of SH-SY5Y cells (F(4,39) = 43.05, P < 0.001), largely when administered-during cell differentiation (F(4,39) = 51.86, P < 0.001). A well-defined dose-response curve was constructed for imidacloprid on day 4 (R2 = 0.945, EC50 = 0.14 mM). During differentiation, either imidacloprid or acetamiprid dose-dependently caused neurite branch retraction on day 3, likely because of oxidative stress, to the extent that cells turned into spheres without neurites after 7-day treatment. Despite their apparent safety, the neurodevelopmental vulnerability of SH-SY5Y neurons to the chronic exposure to imidacloprid and to a lesser extent to acetamiprid points to a neurotoxic risk for humans.

Giant ovarian cystadenocarcinoma in an adult patient, a rare finding in modern times.

Ovarian cancer (OC) is one of the most commonly diagnosed cancers among women. Regretfully due to its a broad spectrum of clinical behavior and challenging diagnosis most cases are diagnosed at a late stage. On rare occasions, these tumors can grow to massive sizes if left untreated, worsening the prognosis of the patient. Thanks to the advancement of medicine and diagnostic techniques, these rare cases are less frequent. Timely detection and surgery could avoid all these potentially troublesome scenarios. We report the case of a 64-year-old female with a giant 13 kg high-grade papillary serous ovarian cystadenocarcinoma, the tumor grew during a four year period and was adequately treated with surgery and is under close follow up with the oncologist. To our knowledge, this is the first case of a giant ovarian cystadenocarcinoma ever reported in Ecuador.

Prenatal gunshot wound, a rare cause of maternal and fetus trauma, a case report.

BACKGROUND: Gunshot wounds in pregnant women, although rare, represent an important cause of fetal and maternal mortality. Understanding the mechanism of injury is essential to identify the possible injuries and to adequately manage the complexity of these emergency scenarios. CASE PRESENTATION: We present a case of a 27-year-old woman and her 37 week fetus who were the victims of a gunshot wound. The trajectory of the bullet injured not only the mother but also the developing fetus. An emergency c-section was performed and the bullet was removed from the infant's abdomen. The two patients fully recovered and on follow up controls both patients are doing well. CONCLUSIONS: Trauma events, and particularly gunshot wounds in pregnant women requires promptly and adequate intervention. Coordinated efforts from multidisciplinary clinical teams are needed as well as the appropriate training in maternal and infant resuscitative measures and surgical techniques.

Subcapsular liver hematoma after extracorporeal shock wave lithotripsy, a rare complication.

Extracorporeal shock wave lithotripsy (ESLW) is a common and relatively safe procedure, with a high success rate and low adverse effects. Complications of this procedure are usually limited to the kidneys and almost always resolve spontaneously. However, on rare occasions, a frightening complication, as a hematoma of a parenchymatous organ, such as the liver can appear. Abdominal pain is usually the only manifestation when a liver hematoma develops, and should always be investigated, as this dreaded complication can put the patient's life at risk. Conservative treatment is usually the treatment of choice, nevertheless, on severe cases that do not respond to conservative measures, surgery or embolization is necessary. We report the case of a 38-year-old male who underwent ESWL, 12 h after the procedure he presented to the emergency room with severe abdominal pain, after complementary exams a liver hematoma was detected. Following conservative management, the patient fully recovered.

Colonic perforation due to amebiasis, a rare and lethal complication.

Amebiasis is still a major healthcare concern, especially in developing countries like Ecuador. The lack of sanitary control and hygiene measures make parasites infections still a burden for patients and physicians. Despite infections due to this parasites are usually mild, severe infections and fatal outcomes although rare still occur. Bowel perforation is a rare complication of amebiasis, and unfortunately, it continues to be almost fatal. We present a case of an Ecuadorian patient who presented to the emergency room with an acute abdomen, despite adequate surgery and critical care, the patient regrettably died. Bowel perforation due to Entamoeba histolytica was the final diagnosis.

Decision support algorithm for the selection of analytical methods in organic compounds detection for future extraterrestrial exploratory missions.

The search for biotic compounds beyond planet Earth is among the highest priorities in space discovery. We sought to detect organic molecules in various earth soil samples collected from environments with extreme climatic conditions comparable to extraterrestrial environments using HPLC, spectrophotometry (UV, IR), GC/MS and vertical gel electrophoresis analyses systems. The applied methods in the study were compared using a self-developed software tool that was designed to evaluate the viability of instrument selection for organics detection in any given exploratory mission. The tool was created to aid further development and refinements of rapidly improving technological solutions such as MCE and MS instruments. Such comparison can be done quickly and easily, acting as a decision support tool when choosing between analytical methods during any planning phase of future exploration missions led by space agencies (i.e., NASA).

Laparoscopic approach to non-communicating intestinal duplication cyst in adult.

Intestinal duplication is rare congenital pathology generally diagnosed in infancy or early childhood. Presentation in adults is extremely rare. We present a case of a 36-year-old otherwise healthy female patient without any past medical history. She presented with a month of lower abdominal pain, clinical examination revealed pain and a palpable mass in the lower abdomen. CT scan confirmed a cystic mass dependent of the cecum wall and colonoscopy showed an extrinsic compression of the lateral wall of the ascending colon. However, the mucosa and the rest of the colon appeared normal. Since pain became more intense laparoscopy was performed and a cystic mass dependent on the cecum was discovered. Laparoscopic hemicolectomy was performed. Patient underwent full recovery. On follow up controls, pathology described intestinal cystic duplication of the colonic wall.

Selective attachment of F-actin with controlled length for developing an intelligent nanodevice.

Development of the nanodevice that myosin-coated beads "walk" on actin filaments (F-actin) tracks for in vitro nanotransportation was hindered due to the difficulty of assembling large-area well-orientated F-actin tracks on the surface. In this work, we present a selective attachment of F-actin with controlled length on a patterned surface by employing biotinylated capped protein gelsolin as intermediate anchoring bridge. A patterned streptavidin layer was formed via coupling with a biotin layer that photo-actively attached to an amine-functionalized glass surface. The patterned film was found stable and homogenous compared to that obtained by microcontact printing method, according to the profiling with fluorescence microscopy. By a secondary blocking process, non-specific binding of F-actin to the patterned surface through electrostatic adsorption can be resisted. The length variation of F-actin as a function of gelsolin concentration was also investigated, implying that F-actin is appropriately of 2.5 µm in average length once F-actin/gelsolin molar ratio is 4:1. Finally, the selective attachment of F-actin was well characterized with quantifying the number of attached F-actin per unit area in the patterned areas over that in blocked areas. The density of F-actin was estimated at c.a. 2 µm(2) per actin filament molecule so that the distance between one another actin filament is estimated as c.a. 1.41-1.97 µm. The unique properties of F-actin, e.g. well flexibility or electrical conductivity, make it feasible to lay them down and form unidirectional aligned tracks by fluidic flow or electrical field. This may open a possibility for the long-distant movement of myosin-coated beads, offering a novel discipline for the development of micro-biochip in vitro.

Comparative effects of virginiamycin supplementation on characteristics of growth-performance, dietary energetics, and digestion of calf-fed Holstein steers.

Two trials were conducted to evaluate the influence of virginiamycin supplementation on growth performance and digestive function. In trial 1, 144 Holstein steer calves (119 kg of initial BW) were used in a 340-d trial. Treatments were (DM basis) 1) control (no antibiotic); 2) 16 mg/kg of virginiamycin; 3) 22.5 mg/kg of virginiamycin; and 4) 28 mg/kg of monensin. Calves received a steam-flaked corn-based growing diet for the first 112 d and thereafter were fed a finishing diet from d 112 until slaughter. Steers were divided into 2 BW groups (lighter-half and heavier-half), and assigned within groups to 24 pens (6 steers/pen). Virginiamycin did not affect (P > 0.20) ADG, but increased 340-d G:F (linear; P = 0.02) and dietary NE (linear; P = 0.04). Improvements in dietary NE were most pronounced during the final 116 d of the feeding period (linear; P = 0.04) that comprised the hot summer months (June through September). Monensin did not affect (P > 0.20) growth performance or dietary NE. Although average initial BW of the lighter-half and heavier-half BW groups differed by only 4 kg, the heavier BW group had greater HCW (13 kg, P = 0.02), LM area (5.4%, P = 0.01), ADG (4.3%, P = 0.02), G:F (2.3%, P = 0.08), and dietary NE (3.2%. P = 0.01) than the lighter BW group. In trial 2, 4 Holstein steers (269 +/- 3.5 kg) with cannulas in the rumen and proximal duodenum were used in a 4 x 4 Latin square design to evaluate treatment effects on digestion. Virginiamycin did not affect (P > 0.20) ruminal digestion of OM, NDF, starch, and N, and microbial efficiency, but tended to linearly increase (P = 0.09) ruminal N efficiency (nonammonia N flow to the small intestine/N intake). Likewise, virginiamycin did not affect (P > 0.20) postruminal or total tract digestion of OM, starch, NDF, and N. Compared with the control diet, monensin depressed total tract digestion of OM (4.2%, P = 0.03) and NDF (29.5%, P = 0.02). There were no treatment effects (P > 0.20) on ruminal pH, or ruminal lactate and total VFA molar concentration. Virginiamycin increased ruminal molar proportion of acetate (quadratic; P = 0.04) and estimated methane production (quadratic; P = 0.07), and decreased propionate molar proportion (quadratic; P = 0.09). Monensin did not affect (P > 0.20) ruminal VFA molar proportions or estimated methane production. We concluded that virginiamycin supplementation can enhance feedlot growth-performance and dietary energetic efficiency of calf-fed Holstein steers.