Depolarization of sperm membrane potential is a common feature of men with subfertility and is associated with low fertilization rate at IVF

STUDY QUESTION. Are significant abnormalities in outward (K+) conductance and resting membrane potential (Vm) present in the spermatozoa of patients undertaking IVF and ICSI and if so, what is their functional effect on fertilization success? SUMMARY ANSWER. Negligible outward conductance (≈5% of patients) or an enhanced inward conductance (≈4% of patients), both of which caused depolarization of Vm, were associated with a low rate of fertilization following IVF. WHAT IS KNOWN ALREADY. Sperm-specific potassium channel knockout mice are infertile with defects in sperm function, suggesting that these channels are essential for fertility. These observations suggest that malfunction of K+ channels in human spermatozoa might contribute significantly to the occurrence of subfertility in men. However, remarkably little is known of the nature of K+ channels in human spermatozoa or the incidence and functional consequences of K+ channel defects. STUDY DESIGN, SIZE AND DURATION. Spermatozoa were obtained from healthy volunteer research donors and subfertile IVF and ICSI patients attending a hospital assisted reproductive techniques clinic between May 2013 and December 2015. In total, 40 IVF patients, 41 ICSI patients and 26 normozoospermic donors took part in the study. PARTICIPANTS/MATERIALS, SETTING, METHODS. Samples were examined using electrophysiology (whole-cell patch clamping). Where abnormal electrophysiological characteristics were identified, spermatozoa were further examined for Ca2+ influx induced by progesterone and penetration into viscous media if sufficient sample was available. Full exome sequencing was performed to specifically evaluate potassium calcium-activated channel subfamily M α 1 (KCNMA1), potassium calcium-activated channel subfamily U member 1 (KCNU1) and leucine-rich repeat containing 52 (LRRC52) genes and others associated with K+ signalling. In IVF patients, comparison with fertilization rates was done to assess the functional significance of the electrophysiological abnormalities. MAIN RESULTS AND THE ROLE OF CHANCE. Patch clamp electrophysiology was used to assess outward (K+) conductance and resting membrane potential (Vm) and signalling/motility assays were used to assess functional characteristics of sperm from IVF and ICSI patient samples. The mean Vm and outward membrane conductance in sperm from IVF and ICSI patients were not significantly different from those of control (donor) sperm prepared under the same conditions, but variation between individuals was significantly greater (P< 0.02) with a large number of outliers (>25%). In particular, in ≈10% of patients (7/81), we observed either a negligible outward conductance (4 patients) or an enhanced inward current (3 patients), both of which caused depolarization of Vm. Analysis of clinical data from the IVF patients showed significant association of depolarized Vm (≥0 mV) with low fertilization rate (P= 0.012). Spermatozoa with electrophysiological abnormities (conductance and Vm) responded normally to progesterone with elevation of [Ca2+]i and penetration of viscous medium, indicating retention of cation channel of sperm (CatSper) channel function. LIMITATIONS, REASONS FOR CAUTION. For practical, technical, ethical and logistical reasons, we could not obtain sufficient additional semen samples from men with conductance abnormalities to establish the cause of the conductance defects. Full exome sequencing was only available in two men with conductance defects. WIDER IMPLICATIONS OF THE FINDINGS. These data add significantly to the understanding of the role of ion channels in human sperm function and its impact on male fertility. Impaired potassium channel conductance (Gm) and/or Vm regulation is both common and complex in human spermatozoa and importantly is associated with impaired fertilization capacity when the Vm of cells is completely depolarized.

Nitrogen-15 and Oxygen-18 Natural Abundance of Potassium Chloride Extractable Soil Nitrate Using the Denitrifier Method

In agroecosystems, most isotopic investigations of NO3- involve the use of tracers that are artificially enriched in 15N. Although the dual isotope composition of NO3-— d15N and d18O is especially beneficial for understanding the origin and fate of NO3-, its use for KCl-extractable soil NO3- has been hampered by the lack of a suitable analytical technique. Our objective was to test whether the denitrifier method, whereby NO3- is reduced to N2O before mass spectrometric analysis, can be used to determine the N and O isotopic composition of NO3- from 2 M KCl soil extracts. Several internationally accepted NO3- standards were dissolved in 2 M KCl, the conventional extractant for soil inorganic N, and inoculated with the bacterial strain Pseudomonas aureofaciens (ATCC no. 13985). The standard deviation of the NO3- standards analyzed did not exceed 0.2‰ for d15N and 0.3‰ for d18O values. After appropriate corrections, differences between our measured and consensus d15N and d18O values of standard NO3- generally were within the standard deviations given for the consensus values. Both d15N and d18O values were reproducible among separate analytical runs. The method was also tested on genuine 2 M KCl extracts from unfertilized and fertilized soils. Depending on N fertilization, the soils had distinct d15N and d18O values, which were attributed to amendment with NH4NO3 fertilizer. Hence, our data indicate that the denitrifier method provides a fast, reliable, precise, and accurate way of simultaneously analyzing the natural abundances of 15N and 18O in KCl-extractable soil NO3-.

Genetic analysis of potassium use efficiency in Brassica oleracea

Potassium (K) fertilizers are used in intensive and extensive agricultural systems to maximize production. However, there are both financial and environmental costs to K-fertilization. It is therefore important to optimize the efficiency with which K-fertilizers are used. Cultivating crops that acquire and/or utilize K more effectively can reduce the use of K-fertilizers. The aim of the present study was to determine the genetic factors affecting K utilization efficiency (KUtE), defined as the reciprocal of shoot K concentration (1/K(shoot)), and K acquisition efficiency (KUpE), defined as shoot K content, in Brassica oleracea. Genetic variation in K(shoot) was estimated using a structured diversity foundation set (DFS) of 376 accessions and in 74 commercial genotypes grown in glasshouse and field experiments that included phosphorus (P) supply as a treatment factor. Chromosomal quantitative trait loci (QTL) associated with K(shoot) and KUpE were identified using a genetic mapping population grown in the glasshouse and field. Putative QTL were tested using recurrent backcross substitution lines in the glasshouse. More than two-fold variation in K(shoot) was observed among DFS accessions grown in the glasshouse, a significant proportion of which could be attributed to genetic factors. Several QTL associated with K(shoot) were identified, which, despite a significant correlation in K(shoot) among genotypes grown in the glasshouse and field, differed between these two environments. A QTL associated with K(shoot) in glasshouse-grown plants (chromosome C7 at 62 center dot 2 cM) was confirmed using substitution lines. This QTL corresponds to a segment of arabidopsis chromosome 4 containing genes encoding the K(+) transporters AtKUP9, AtAKT2, AtKAT2 and AtTPK3. There is sufficient genetic variation in B. oleracea to breed for both KUtE and KUpE. However, as QTL associated with these traits differ between glasshouse and field environments, marker-assisted breeding programmes must consider carefully the conditions under which the crop will be grown.

Mapeamento das formas do relevo para estimativa de custos de fertilização em cana-de-açúcar

A modelagem matemática associada ao conhecimento da variabilidade dos atributos do solo e mapeamento das formas do relevo pode auxiliar no manejo da fertilidade do solo em usinas sucroalcooleiras. O presente trabalho teve como objetivo avaliar o uso da geoestatística e da modelagem matemática na estimativa de custos de fertilização, em diferentes formas do relevo. em uma área de 200 ha, foram identificadas duas formas de relevo, uma côncava e outra convexa, sendo os solos coletados nos pontos de cruzamento de uma malha, com intervalos regulares de 50 m, perfazendo um total de 623 pontos. As amostras foram submetidas a análises químicas, e, posteriormente, os dados foram avaliados por meio da estatística descritiva, geoestatística e modelagem matemática. Os resultados mostraram que, quando as formas do relevo são incorporadas às análises geoestatística e de modelagem matemática, ocorre aumento na eficiência de aplicação do calcário, fósforo e potássio no solo.

Aspectos biológicos e dano de Diatraea saccharalis (fabr., 1794) (Lepidoptera: Pyralidae) em sorgo cultivado sob diferentes doses de nitrogênio e potássio

O presente trabalho teve como objetivo avaliar a influência da adubação da cultura do sorgo Sorghum bicolor (L.) Moench, na biologia da broca da cana-de-açúcar Diatraea saccharalis (Fabr., 1794) (Lepidoptera: Pyralidae). Para isso foi utilizado sorgo da cultivar Rubi-Asgrow, plantado em vasos utilizando-se diferentes doses de fertilizantes e mantidos em casa-de-vegetação. Os tratamentos utilizados (doses de NK) foram: N1 = 0-200 ppm; N2 = 50-200 ppm; N3 = 100-200 ppm; N4 = 200-200 ppm; N5 = 400-200 ppm; K1 = 200-0 ppm; K2 = 200-50 ppm; K3 = 200-100 ppm; K4 = 200-200 ppm; e K5 = 200-400 ppm. de modo geral, pode-se concluir que doses de 50 a 200 ppm de N promoveram o desenvolvimento normal das larvas de D. saccharalis, sendo que as menores porcentagens de dano foram verificadas nas menores doses; para o potássio, quanto maior a dose, menor foi o dano causado pelas lagartas, apesar de favorecer o desenvolvimento da mesma.

Macronutrients in Marandu Palisade Grass as Influenced by Lime, Slag, and Nitrogen Fertilization

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Impacts of limestone and nitrogen top dressing application on the potassium content in the soil profile and marandu-grass leaf concentration

Objetivou-se avaliar os efeitos de nitrogênio (0, 100, 200, 300 e 400 kg/ha.ano), com ou sem aplicação de calcário dolomítico em cobertura, sobre o teor de potássio (K) do solo nas profundidades de 0 a 5; 5 a 10; e 10 a 20 cm e a concentração de potássio na planta. O experimento foi realizado em Latossolo Vermelho distroférrico de relevo suavemente ondulado, disposto em esquema fatorial 5 × 2, em blocos casualizados, com quatro repetições. A adubação nitrogenada promoveu aumento linear na concentração de potássio na planta e no teor desse mineral na camada de 0 a 5 cm do solo. Ocorreu efeito de inibição competitiva entre a absorção de potássio e cálcio e entre potássio e magnésio. Embora uma fração significativa de potássio retorne à camada superficial do solo pelo resíduo de forragem, em decorrência da maior produção de massa, o aumento da concentração de potássio na planta nas mais altas doses de nitrogênio comprova a importância do suprimento de potássio para a planta, por intensificar a massa de forragem.

Seed germination and seedling development of white oat affected by silicon and phosphorus fertilization

Os fertilizantes silicatados tem sido cada vez mais usados na agricultura devido a inúmeros benefícios, tais como correção da acidez de solos tropicais e efeitos positivos no desenvolvimento de gramíneas. A disponibilidade de nutrientes e a nutrição de plantas desempenham papel importante na produção de sementes e podem influenciar a qualidade fisiológica de sementes de aveia-branca (Avena sativa L.). Avaliou-se a germinação de sementes e o desenvolvimento de plântulas de aveia-branca em função da adubação com silício e fósforo. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 2 x 4, com seis repetições. Os tratamentos consistiram de 20 e 200 mg dm-3 de P2O5, aplicados na forma de superfosfato triplo, combinados com 0, 150, 300 e 450 mg dm-3 de Si na forma de silicato de potássio. O experimento foi realizado em casa de vegetação, conduzindo-se sete plantas por vaso, com capacidade para 15 L de terra. As panículas foram colhidas e debulhadas manualmente e, as sementes, armazenadas em sacos de papel em condições normais de ambiente. As sementes foram avaliadas quanto ao teor de água, massa de sementes, germinação, condutividade elétrica, comprimento e massa da matéria seca de plântulas. Sementes de aveia-branca com qualidade superior são produzidas com 20 mg dm-3 de P2O5, independente da dose de Si. Sementes com maior germinação e vigor são obtidas com 300 e 450 mg dm-3 de K2SiO3, respectivamente. Os comprimentos da raiz e total das plântulas foram inferiores nas doses de Si até 300 kg ha-1, porém a dose de fósforo somente afetou o desenvolvimento das plântulas de maneira distinta quando aplicada junto com a maior dose de silício.

Sugarcane bagasse ash as a potential quartz replacement in red ceramic

Sugarcane bagasse ash (SCBA) is an industrial waste that contains silicon and aluminum oxides as the major components and iron, calcium, magnesium, and potassium oxides as the main minor components. In this paper, SCBA from one Brazilian factory was characterized and tested for its influence on the ceramic properties of clay/ash ceramic probes. Prismatic probes were pressed (18 MPa) using a ceramic mass mixed with 0%, 5%, 8%, and 10% ash. The probes were fired at temperatures between 800 degrees and 1200 degrees C. X-ray diffraction, X-ray fluorescence, thermal analysis (differential thermal analysis, thermo-gravimetric analysis/differential thermogravimetric analysis), and tests for texture (particle-size analysis), flexural strength, and linear shrinkage were carried out to characterize the samples. The results showed that the amount of ash to be incorporated will depend on mainly the composition of clay but also ash, and indicated that the clay used in this work can incorporate up to 10% weight of ash to produce solid bricks. The results also showed an improvement in ceramic/ash properties up to sintering temperatures higher than 1000 degrees C.

Produtividade e qualidade de cana-de-açúcar cultivada em solo tratado com lodo de esgoto, vinhaça e adubos minerais

O uso agrícola de resíduos orgânicos, de origem agrícola, urbana ou industrial, é uma interessante alternativa de disposição, permitindo a reciclagem de nutrientes (NPK) nos ecossistemas. Este trabalho avaliou o efeito da aplicação de lodo de esgoto como fonte de N e de vinhaça como fonte de K comparado ao uso de fontes minerais desses nutrientes sobre a produtividade e variáveis agroindustriais da cana-de-açúcar, por dois anos consecutivos (cana-planta e cana-soca). O experimento foi conduzido em Latossolo Vermelho-Amarelo distrófico típico, em Pontal - SP, e a variedade de cana-de-açúcar avaliada foi a SP 81-3250. Utilizou-se de esquema fatorial 3x2x2+1, ou seja, três tipos de resíduos (lodo de esgoto + KCl; vinhaça + uréia, e lodo de esgoto + vinhaça); dois modos de aplicação (na linha de plantio ou em área total); duas doses (100 e 200% do N e K necessários à cultura) e um tratamento adicional com adubação mineral, sendo os tratamentos distribuídos na área em blocos ao acaso, com três repetições. Foram avaliadas a produtividade e as variáveis agroindustriais (°brix, pol no caldo, fibra, pureza, pol na cana, AR e ATR). As produtividades de colmo e de açúcar para cana-planta foram mantidas quando N e K foram fornecidos pelo lodo de esgoto e vinhaça, respectivamente. A cana-soca apresentou maior produtividade de colmo e de açúcar quando foram utilizados os resíduos separadamente, complementados com fontes minerais. Quanto ao modo de aplicação, não foram observadas diferenças significativas para as variáveis analisadas.

Fermentation, losses, and aerobic stability of sugarcane silages treated with chemical or bacterial additives

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Greenhouse gas balance due to the conversion of sugarcane areas from burned to green harvest in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spatial variability of soil attributes and sugarcane yield in relation to topographic location

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Potassium leaching from millet straw as affected by rainfall and potassium rates

Pearl millet (Penisetum glaucum) is an interesting species to be used as cover crop in tropical areas, showing a high ability in potassium uptake. Potassium (K) is not linked to organic compounds in the plant, and can easily be released from decaying straw becoming available for subsequent crops. This experiment evaluated K leaching from millet straw grown under potassium rates (0, 100, 200, and 300 mg dm-3), and submitted to five levels of simulated rain (5, 10, 20, 40, and 80 mm). Plants were grown in soil filled pots in a greenhouse. On the 50th day after emergence, the plants were desiccated with glyphosate. Artificial rain was applied over the straw. Potassium deficiency speeds up millet dehydration after herbicide application and increases lightly rain water retention in the straw. The amount of K leached right after plant desiccation is correlated with the residue nutrient content and can be as high as 64 kg ha-1 considering a mulch of 8 t ha -1. Although well-nourished millet plants release considerable amounts of K with the first rains, a large percentage of the nutrient is still retained in the straw. Copyright © Taylor & Francis, Inc.

Aplicação de silicato de potássio e crescimento foliar da cana-de-açúcar

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)