Development of a rapid loop-mediated isothermal amplification (LAMP) assay for visual detection of porcine parvovirus (PPV) and its application.

Porcine parvovirus (PPV) infection is one of the most important causes of reproductive failure in pigs impacting the piggery industry globally with huge economic losses. A cost-effective, simple, rapid, specific, and sensitive method is critical for monitoring PPV infection on pig farms. The main aim of the present study was to develop and evaluate a loop-mediated isothermal amplification (LAMP) assay for rapid visual detection of porcine parvovirus (PPV) in pigs. A set of six LAMP primers including two outer primers, two inner primers, and two loop primers were designed utilizing the conserved region of capsid protein VP2 gene sequences of PPV and was applied for detection of PPV from porcine samples. Time and temperature conditions for amplification of PPV genes were optimized to be 30 min at 63 °C. The developed assay was ten-fold more sensitive than conventional PCR with analytical sensitivity of 20 pg and 200 pg, respectively. This is the first report of detection of PPV by LAMP assay from India. The assay did not cross-react with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), or classical swine fever virus (CSFV). The LAMP assay was assembled into a LAMP assay kit of 20 reactions and was validated in different laboratories in India. The newly developed LAMP assay was proved to be a specific, sensitive, rapid, and simple method for visual detection of PPV which does not require even costly equipments for performing the test. It complements and extends previous methods for PPV detection and provides an alternative approach for detection of PPV.

Real-Time Quantitative and Ion-Metal Indicator LAMP-Based Assays for Rapid Detection of Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum is one of the most devastating and cosmopolitan plant pathogens. Rapid detection of S. sclerotiorum can provide growers an advantage in knowing what control measures should be taken to minimize crop damage and financial losses caused by it. Loop-mediated isothermal amplification (LAMP) is a fast, sensitive, and specific nucleic acid amplification method that does not require a thermal cycler. This study aimed to develop a LAMP-based assay for the specific detection of S. sclerotiorum (Ss-LAMP). A real-time quantitative LAMP reaction (Ss-qLAMP) and a calcein ion indicator-based LAMP reaction (Ss-cLAMP) were designed, optimized, and tested on fungi, plant, and soil samples. The Ss-LAMP reactions were very specific and sensitive. Applying the artificially inoculated soil samples with DNA purified by five protocols in the Ss-qLAMP reaction, it was possible to detect and quantify the pathogen DNA, regardless of the extraction protocol. Naturally infected soybean tissues had the pathogen detected by Ss-cLAMP directly in the reaction tube with no DNA extraction requirement. The assays should be applicable for many types of samples, such as soil, spore traps, and plant tissues from several crops, with no requirement for DNA extraction. The Ss-LAMP reactions took less than 1 h to complete, and they can be made directly in the field with real-time quantitative results (Ss-qLAMP) or qualitative naked-eye visual results (Ss-cLAMP). Results were obtained with 10 pg of DNA or 10 ng of crude mycelium, suggesting a detection limit close to a single DNA copy. Ss-LAMP reactions will allow rapid and accurate diagnosis of S. sclerotiorum and assist in pathogen management and control.

A simple protocol for the visual discrimination of natural cyclodextrins in aqueous solution using perichromic probes.

Three synthesized compounds, 4-(4-nitrostyryl)phenol, 2,6-dibromo-4-(2,4-dinitrobenzylideneamino)phenol and 2,6-dichloro-4-(2,4-dinitrobenzylideneamino)phenol, were deprotonated to generate the perichromic dyes 2b, 3b and 4b, respectively. These dyes were used as probes to investigate the micropolarity of natural cyclodextrins (CyDs) and it was observed that they interact differently with the CyDs according to the molecular structure of the dye and the diameter of the CyD. The solvatochromic bands of the dyes that interacted with the CyDs were bathochromically shifted, suggesting that the probes were transferred to the hydrophobic interior of the CyD in aqueous solution. Dyes 2b and 4b were found to be very selective for α-CyD and γ-CyD, respectively, while ß- and γ-CyD changed the color of the solution of compound 3b. These dyes were then successfully used in a simple assay that allows the naked-eye discrimination of natural CyDs in aqueous solution, without the need for expensive techniques.

Limits of visual detection for finasteride polymorphs in prepared binary mixtures: analysis by X-ray powder diffraction.

Finasteride (FNT) is a drug that inhibits human enzyme type II 5α-reductase that metabolizes testosterone into dihydrotestosterone. There are two enantiotropic polymorphs with known crystal structure: designated as forms I and II. Identification and control of these polymorphic forms in mixtures can be performed using X-ray powder diffraction (XRPD) data and Rietveld method (RM). As experimental conditions may limit the detection of minority phases in mixtures, it is interesting to show what are these limits for some usual and one high-resolution equipment. So, in this work, we discuss the parameters to find the limit of the detection in binary mixtures of forms I and II of FNT according to each experimental condition. The samples analyzed were binary mixtures prepared with anhydrous polymorphs of the drug FNT. These samples were measured in four diffractometers with different experimental condition. These equipments represent the main resolutions generally used for drug analysis by XRPD. For the development of this work, a batch of form I was obtained pure, and another batch with forms I and II was used to obtain pure form II by heat treatment. Depending on the experimental condition, the polymorphs could be detected in a proportion as low as 0.5 wt%. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 103:3567-3575, 2014.

Percepción de movimiento coherente en pacientes con glaucoma y estrabismo / Coherent motion perception in patients with glaucoma and strabismus

Se realizó una investigación analítica y prospectiva, de casos y controles, de los niños con glaucoma o estrabismo, o ambos, atendidos en la consulta de Oftalmología del Hospital Infantil Sur de Santiago de Cuba, de enero a junio del 2011, a fin de evaluar la percepción visual del movimiento coherente en ellos. El grupo de casos incluyó 72 pacientes escogidos de forma aleatoria, y el de controles, igual cantidad de integrantes que presentaban toda su capacidad visual. Se determinó que los niños con afecciones en la visión cometían más errores, omisiones y menos aciertos, además de ser menos rápidos en la solución de los ensayos orientados, pues tuvieron mayor tiempo de reacción para ejecutarlos.

A prospective analytic case-control study was carried out in children with glaucoma or strabismus or both, treated at the Department of Ophthalmology of the Southern Children Hospital in Santiago de Cuba, from January to June 2011, with the purpose of assessing the visual perception of coherent motion in them. The case group included 72 randomly selected patients, and control group with the same number of members who had all their vision. It was determined that children with visual disorders made more errors, omissions and fewer hits, besides being less rapid in solving oriented tests, as they had longer reaction time to do them.

Comparison of cap lamp and laser illumination for detecting visual escape cues in smoke.

The Illuminating Engineering Society of North America reports that an underground mine is the most difficult environment to illuminate (Rea, 2000). Researchers at the U.S. National Institute for Occupational Safety and Health (NIOSH) Office of Mine Safety and Health Research (OMSHR) are conducting ongoing studies designed to explore different lighting technologies for improving mine safety. Underground miners use different visual cues to escape from a smoke-filled environment. Primary and secondary escapeways are marked with reflective ceiling tags of various colors. Miners also look for mine rail tracks. The main objective of this paper is to compare different lighting types and ceiling tag colors to differentiate what works best in a smoke-filled environment. Various cap lamps (LED and incandescent) and lasers (red, blue, green) were compared to see which options resulted in the longest detection distances for red, green and blue reflective markers and a section of mine rail track. All targets advanced toward the human subject inside of a smoke-filled room to simulate the subject walking in a mine environment. Detection distances were recorded and analyzed to find the best cap lamp, laser color and target color in a smoke environment. Results show that cap lamp, laser color and target color do make a difference in detection distances and are perceived differently based on subject age. Cap lamps were superior to lasers in all circumstances of ceiling tag detection, with the exception of the green laser. The incandescent cap lamp worked best in the simulated smoke compared to the LED cap lamps. The green laser was the best color for detecting the tags and track compared to the red and blue lasers. The green tags were the easiest color to detect on the ceiling. On average, the track was easier for the subjects to detect than the ceiling tags.

Comparison of cap lamp and laser illumination for detecting visual escape cues in smoke.

The Illuminating Engineering Society of North America reports that an underground mine is the most difficult environment to illuminate (Rea, 2000). Researchers at the U.S. National Institute for Occupational Safety and Health (NIOSH) Office of Mine Safety and Health Research (OMSHR) are conducting ongoing studies designed to explore different lighting technologies for improving mine safety. Underground miners use different visual cues to escape from a smoke-filled environment. Primary and secondary escapeways are marked with reflective ceiling tags of various colors. Miners also look for mine rail tracks. The main objective of this paper is to compare different lighting types and ceiling tag colors to differentiate what works best in a smoke-filled environment. Various cap lamps (LED and incandescent) and lasers (red, blue, green) were compared to see which options resulted in the longest detection distances for red, green and blue reflective markers and a section of mine rail track. All targets advanced toward the human subject inside of a smoke-filled room to simulate the subject walking in a mine environment. Detection distances were recorded and analyzed to find the best cap lamp, laser color and target color in a smoke environment. Results show that cap lamp, laser color and target color do make a difference in detection distances and are perceived differently based on subject age. Cap lamps were superior to lasers in all circumstances of ceiling tag detection, with the exception of the green laser. The incandescent cap lamp worked best in the simulated smoke compared to the LED cap lamps. The green laser was the best color for detecting the tags and track compared to the red and blue lasers. The green tags were the easiest color to detect on the ceiling. On average, the track was easier for the subjects to detect than the ceiling tags.

Evaluación de la percepción del movimiento en niños y niñas disléxicos / Evaluation of movement perception in dyslexic boys and girls

Se aplicó una tarea de detección visual del movimiento coherente, confeccionada en el Centro de Neurociencias y Procesamiento de Imágenes y Señales de la Universidad de Oriente, en Santiago de Cuba, mediante la cual se determinó que los escolares disléxicos estudiados podían detectar el movimiento visual, pero cometían muchos errores y omisiones, indicativos de un ligero debilitamiento del proceso de detección visual. También se observaron deficiencias perceptivas en ellos, relacionadas con el incremento de la densidad de los puntos que se movían en los paneles y su disminución porcentual, así como también con el aumento del tiempo de respuesta, reveladores de una reducción de la sensibilidad a la percepción visual del movimiento en estos alumnos con dislexia del desarrollo.

A task of visual detection of the coherent movement, made in the Neurosciences and Processing of Images and Signs Center from Oriente University was applied in Santiago de Cuba, by means of which it was determined that the dyslexic students studied, could detect the visual movement, but they made many mistakes and omissions, indicative of a slight weakening of the visual detection process. Perceptive deficiencies were also observed in them, related with the increment of the density of the points which moved in the panels and their percentage decrease, as well as with the increase of the response time, showing of a reduction of the sensibility to the visual perception of movement in these students with development dyslexia.