ABSTRACT
Single-molecule techniques are ideally poised to characterize complex dynamics but are typically limited to investigating a small number of different samples. However, a large sequence or chemical space often needs to be explored to derive a comprehensive understanding of complex biological processes. Here we describe multiplexed single-molecule characterization at the library scale (MUSCLE), a method that combines single-molecule fluorescence microscopy with next-generation sequencing to enable highly multiplexed observations of complex dynamics. We comprehensively profiled the sequence dependence of DNA hairpin properties and Cas9-induced target DNA unwinding-rewinding dynamics. The ability to explore a large sequence space for Cas9 allowed us to identify a number of target sequences with unexpected behaviors. We envision that MUSCLE will enable the mechanistic exploration of many fundamental biological processes.
Subject(s)
DNA , High-Throughput Nucleotide Sequencing , Microscopy, Fluorescence , Single Molecule Imaging , High-Throughput Nucleotide Sequencing/methods , Single Molecule Imaging/methods , DNA/chemistry , DNA/genetics , Microscopy, Fluorescence/methods , CRISPR-Associated Protein 9 , Sequence Analysis, DNA/methods , Gene Library , CRISPR-Cas SystemsABSTRACT
Fructans contribute significantly to dietary fiber with beneficial effects on gastrointestinal physiology in healthy individuals and offer a promising approach to treating some diseases. Two experiments (Experiment 1 = rats with normal weight; Experiment 2 = obese rats) were developed to compare the effects of three fructan sources (Cichorium intybus L. Asteraceae, Helianthus tuberosus L. Asteraceae and Agave angustifolia ssp. tequilana Haw, Agavaceae) on body weight change, blood metabolites and fecal bacteria in non-diabetic (ND) and diabetic (D) rats. In Experiment 1 total body weight gain and daily feed intake in D and ND rats decreased (P < 0.05) with supplements of fructan. Only in D rats, blood glucose concentrations, fecal Clostrodium spp. counts, and liver steatosis decreased, while blood HDL concentrations and fecal Lactobacillus spp. and Bifidobacterium spp. counts increased due to fructans. In Experiment 2, total body weight gain and feed intake in ND and D rats were also decreased by fructans. In ND rats, fructan decreased blood glucose concentrations. In D rats, fructans from A. angustifolia ssp. tequilana decreased blood cholesterol and LDL and liver steatosis. For both ND and D rats, fecal Lactobacillus spp. and Bifidobacterium spp. counts were higher (P < 0.05) with fructan supplements.
Subject(s)
Asteraceae/chemistry , Bacteria/drug effects , Body Weight/drug effects , Diabetes Mellitus, Experimental/metabolism , Fructans/pharmacology , Obesity/metabolism , Animals , Blood Glucose/analysis , Cholesterol/blood , Diabetes Mellitus, Experimental/complications , Feces/microbiology , Fructans/isolation & purification , Male , Obesity/complications , Random Allocation , Rats , Rats, Wistar , Triglycerides/blood , Weight Gain/drug effectsABSTRACT
Heliopsis longipes S.F. Blake (Asteraceae: Heliantheae) (chilcuague) is used in Mexican traditional medicine against parasites and to alleviate tooth and muscle pains. Its biocide effect has already been experimentally demonstrated; however, its analgesic action and its action on the nervous system (NS) have not been investigated yet. The objectives of this study were to evaluate the analgesic action of affinin and the H. longipes root ethanol extract, as well as their effects on the NS using an animal model. The ethanol extract was obtained by maceration, and affinin was purified from it through chromatographic techniques. Chemical and thermal analgesia were used to assess their analgesic proprieties. Irwin's test was used to evaluate their stimulating or depressing effects. The ethanol extract and affinin displayed analgesic action similar to ketorolac and stimulating effect comparable to caffeine on the nervous system of adult mice.
Subject(s)
Analgesics, Non-Narcotic/therapeutic use , Asteraceae/chemistry , Nervous System/drug effects , Pain/drug therapy , Phytotherapy , Plant Extracts/therapeutic use , Analgesics, Non-Narcotic/chemistry , Analgesics, Non-Narcotic/isolation & purification , Animals , Behavior, Animal/drug effects , Central Nervous System Depressants/chemistry , Central Nervous System Depressants/isolation & purification , Central Nervous System Depressants/pharmacology , Central Nervous System Stimulants/chemistry , Central Nervous System Stimulants/isolation & purification , Central Nervous System Stimulants/pharmacology , Hot Temperature/adverse effects , Male , Medicine, Traditional , Mexico , Mice , Pain/chemically induced , Pain Measurement , Plant Extracts/chemistry , Plant Roots/chemistry , Polyunsaturated Alkamides/isolation & purification , Polyunsaturated Alkamides/therapeutic use , Time FactorsABSTRACT
The chemical composition of the essential oil of Chysactinia mexicana was analyzed by gas chromatography-mass spectrometry. Seventeen compounds were characterized; eucalyptol (41.3%), piperitone (37.7%), and linalyl acetate (9.1%) were found as the major components. The essential oil of leaves and piperitone completely inhibited Aspergillus flavus growth at relatively low concentrations (1.25 and 0.6 mg/mL, respectively).
Subject(s)
Asteraceae/chemistry , Fungicides, Industrial/pharmacology , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Aspergillus flavus/drug effects , Aspergillus flavus/growth & development , Gas Chromatography-Mass Spectrometry , Plant Leaves/chemistry , Plant Oils/chemistry , Plant Oils/pharmacologyABSTRACT
Entre 1993 a 1999 se realizaron herborizaciones sistemáticas en selva alta perennifolia, acahuales (comunidades secundarias) de diferentes edades y milpas pertenecientes a la comuidad de Lacanhá Chansayab, en la Selva Lacandona. Los especímenes recolectados se identificaron y procesaron como ejemplares de herbario acompañados de información sobre habitat, forma de vida y formas de aprovechamiento. Se recabó información sobre la importancia relativa de estas especies útiles en la estructura de las selvas altas y acahuales del sureste mexicano. Se registraron 485 especies de plantas vasculares, de las cuales el 73,4 por ciento (356 especies) fue reconocido y aprovechado por los lacandones. De un alto porcentaje (77 por ciento) de estas especies se obtuvieron materiales vegetales para satisfacer necesidades de autoconsumo como construcción, alimentos, leña y medicamentos; otros materiales son extraídos con fines comerciales como los requeridos para elaborar artesanías, y venta de fibra de pita y de hojas de palmas. En la estructura de la selva y acahuales destacan 124 especies utilizadas por los lacandones. El uso de estas especies a lo largo de un gradiente sucesional nos muestra el carácter persistente del aprovechamiento del recurso vegetal por esta etnia; así la riqueza de especies utilizada por lacandones se incrementa directamente con el período de descanso de la vegetación