Surface programmed bacteria as photo-controlled NO generator for tumor immunological and gas therapy.

The use of bacteria as living vehicles has attracted increasing attentions in tumor therapy field. The combination of functional materials with bacteria dramatically facilitates the antitumor effect. Here, we presented a rationally designed living system formed by programmed Escherichia Coli MG1655 cells (Ec) and black phosphorus (BP) nanoparticles (NPs). The bacteria were genetically engineered to express tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), via an outer membrane YiaT protein (Ec-T). The Ec-T cells were associated with BP NPs on their surface to acquire BP@Ec-T. The designed living system could transfer the photoelectrons produced by BP NPs after laser irradiation and triggered the reductive metabolism of nitrate to nitric oxide for the in situ release at tumor sites, facilitating the therapeutic efficacy and the polarization of tumor associated macrophages to M1 phenotype. Meanwhile, the generation of reactive oxygen species induced the immunogenic cell death to further improve the antitumor efficacy. Additionally, the living system enhanced the immunological effect by promoting the apoptosis of tumor cells, activating the effect of T lymphocytes and releasing the pro-inflammatory cytokines. The integration of BP NPs, MG1655 cells and TRAIL led to an effective tumor therapy. Our work established an approach for the multifunctional antitumor living therapy.

Impact of nitrogen and phosphorus fertilizers on Cry1Ac protein contents in transgenic cotton / Impacto de fertilizantes de nitrogênio e fósforo nos conteúdos de proteína Cry1Ac em algodão transgênico

Application of different fertilizers to check the efficiency of expression of Bt (Bacillus thuringiensis) gene in one of the leading commercialized crops (cotton) against Lepidopteran species is of great concern. The expression of Cry protein level can be controlled by the improvement of nutrients levels. Therefore, the myth of response of Cry toxin to different combinations of NP fertilizers was explored in three Bt cotton cultivars. Combinations include three levels of nitrogen and three levels of phosphorus fertilizers. Immunostrips and Cry gene(s) specific primer based PCR (Polymerase Chain Reaction) analysis were used for the presence of Bt gene that unveiled the presence of Cry1Ac gene only. Further, the ELISA (enzyme-linked immunosorbent assay) kit was used to quantify the expression of Cry1Ac protein. Under various NP fertilizers rates, the level of toxin protein exhibited highly significant differences. The highest toxin level mean was found to be 2.3740 and 2.1732 µg/g under the treatment of N150P75 kg ha-¹ combination while the lowest toxin level mean was found to be 0.9158 and 0.7641 µg/g at the N50P25 kg ha-¹ level at 80 and 120 DAS (Days After Sowing), respectively. It was concluded from the research that the usage of NP fertilizers has a positive relation with the expression of Cry1Ac toxin in Bt cotton. We recommend using the N150P50 kg ha-1 level as the most economical and practicable fertilizer instead of the standard dose N100P50 kg ha-¹ to get the desired level of Cry1Ac level for long lasting plant resistance (<1.5). The revised dose of fertilizer may help farmers to avoid the cross-resistance development in contradiction of insect pests.

A aplicação de diferentes fertilizantes para verificar a eficiência da expressão do gene Bt (Bacillus thuringiensis) em uma das principais culturas comercializadas (algodão) contra espécies de lepidópteros é uma grande preocupação. A expressão do nível de proteína Cry pode ser controlada pela melhoria dos níveis de nutrientes. Portanto, o mito da resposta da toxina Cry a diferentes combinações de fertilizantes NP foi explorado em três cultivares de algodão Bt. As combinações incluem três níveis de nitrogênio e três níveis de fertilizantes de fósforo. A análise de PCR (reação em cadeia da polimerase) específica para o gene (s) Immunostrips e Cry (s) foi usada para a presença do gene Bt que revelou a presença do gene Cry1Ac apenas. Além disso, o kit ELISA (ensaio de imunoabsorção enzimática) foi usado para quantificar a expressão da proteína Cry1Ac. Sob várias taxas de fertilizantes NP, o nível de proteína de toxina exibiu diferenças altamente significativas. A média do nível mais alto de toxina foi de 2,3740 e 2,1732 µg / g sob o tratamento da combinação N150P75 kg ha-¹, enquanto a média do nível mais baixo de toxina foi de 0,9158 e 0,7641 µg / g no nível de N50P25 kg ha-¹ em 80 e 120 DAS (dias após a semeadura), respectivamente. Concluiu-se com a pesquisa que o uso de fertilizantes NP tem relação positiva com a expressão da toxina Cry1Ac no algodão Bt. Recomendamos o uso do nível de N150P50 kg ha-¹ como o fertilizante mais econômico e praticável em vez da dose padrão N100P50 kg ha-¹ para obter o nível desejado de nível de Cry1Ac para resistência de planta de longa duração (<1,5). A dose revisada de fertilizante pode ajudar os agricultores a evitar o desenvolvimento de resistência cruzada em contradição com as pragas de insetos.

The Relationship Between Periodontal Disease and Nutrient Intake in Korean Adults: The Korea National Health and Nutrition Examination Survey (KNHANES VII) from 2016-2018.

PURPOSE: To examine the association between the intake of various nutrients (phosphorus, riboflavin, thiamine, niacin, vitamin C, calcium, protein, carbohydrates, and fat) and the prevalence of periodontal disease in Korean adults. MATERIALS AND METHODS: The data used for analysis were obtained from the 7th Korea National Health and Nutrition Examination Survey (2016-2018). Data from 12,689 adults aged ≥ 19 years who had a periodontal examination were analysed. Data were analysed using the Chi2 and t-tests. Multiple regression analysis was used to assess the association between the selected nutrients and periodontal diseases. RESULTS: After adjusting for sex, age, income, body mass index, diabetes, smoking, alcohol consumption, and toothbrushing frequency, a statistically significant relationship between phosphorus, carbohydrate, and fat intake and the risk of periodontal disease was identified by multiple logistic regression analysis (odds ratio [OR]: 0.80, 95% confidence interval [CI]: 0.66-0.97; OR: 0.85, 95% CI: 0.70-0.98, OR: 1.41, 95% CI: 1.13-1.75, respectively). CONCLUSION: Phosphorus, carbohydrates and fat were associated with periodontal disease. Therefore, the improvement of diet should be emphasised to prevent and manage periodontal disease. Further research is needed based on various nutrients related to periodontal disease in the future.

Effects of different combinations of N, P and K at different time interval on vegetative, reproductive, yield and quality traits of mango (Mangifera Indica. L) cv. Dusehri / Efeitos de diferentes combinações de N, P e K em diferentes intervalos nas características vegetativas, reprodutivas, produtivas e de qualidade da manga (Mangifera indica. L) cv. Dusehri

The experiment was carried out on mango cv. Dusehri to investigate the effect of N, P and K fertilizers on vegetative, reproductive growth, yield and fruit quality. Eight different fertilizer combinations such as T1 (control), T2 (N), T3 (P), T4 (K), T5 (NP), T6 (NK), T7 (PK) and T8 (NPK) were used. Individual or combine fertilizer application of N (1000 g), P (750 g) and K (750 g) were applied during growing season in February and August. All the treatments significantly influenced on vegetative growth, flowering, fruiting, yield and other physiochemical attributes of mango as compared to control. Least effect was observed with individual fertilizer application while combine fertilizer treatments enhanced most of the investigated parameters. Especially, qualitative traits showed non significant differences between treated and untreated mango trees. However, among the different treatments T8 (NPK) showed significance for fruiting aspects such as maximum size of growth flushes (177.51 mm), total number of panicles/tree (845), total number of flowers/panicle (974), sex ratio (69.18%), fruit retention (13.85%), total number of fruits/tree (379), yield (82 kg/tree), fruit weight (197.5 g), pulp weight (135.5 g) and physiochemical parameters namely TSS (24.53), Vit. C (57.63 mg/100 mL) and total sugar (20.84%). In general, combine application of NPK (T8) were the most effective in enhancing fruiting aspects, yield, physiochemical characteristics as well as improved fruit quality of mango trees.

O experimento foi realizado em manga cv. Dusehri para investigar o efeito dos fertilizantes N, P e K no crescimento vegetativo, reprodutivo, produtividade e de qualidade do fruto. Foram utilizadas oito combinações diferentes de fertilizantes: T1 (controle), T2 (N), T3 (P), T4 (K), T5 (NP), T6 (NK), T7 (PK) e T8 (NPK). Cada tratamento de N(1.000 g), P (750 g) e K (750 g) foi aplicado duas vezes durante a estação de crescimento em fevereiro e agosto. Todos os tratamentos influenciaram significativamente o crescimento vegetativo, floração, frutificação, produtividade e outros atributos físico-químicos da manga em relação ao controle. Menos efeito foi observado com a aplicação individual de fertilizante, enquanto os tratamentos combinados aumentaram a maioria dos parâmetros investigados. Especialmente as características qualitativas mostraram diferenças não significativas entre mangueiras tratadas e não tratadas. No entanto, entre os diferentes tratamentos, T8 (NPK) apresentou significância para aspectos de frutificação, como tamanho máximo de folgas de crescimento (177,51 mm), número total de panículas/árvore (845), número total de flores/panícula (974), razão sexual (69,18%), retenção de frutos (13,85%), número total de frutos/árvore (379), produção (82 kg/árvore), peso do fruto (197,5 g) e peso da polpa (135,5 g), além de parâmetros físico-químicos, como TSS (24,53), vitamina C (57,63 mg/100 mL) e açúcar total (20,84%). Em geral, a aplicação combinada de NPK (T8) foi a mais eficaz no aprimoramento dos aspectos de frutificação, produtividade, características físico-químicas, além da melhoria da qualidade dos frutos das mangueiras.

Effects of nitrogen and phosphorus availability on the early growth of two congeneric pairs of savanna and forest species / Efeitos da disponibilidade de nitrogênio e fósforo no crescimento inicial de dois pares congenéricos de espécies de savana e floresta

In the tropical region, savannas and seasonal forests, both highly diverse biomes, occur side by side, under the same climate. If so, that mosaic cannot be explained solely by climatic variables, but also by fire, water availability and soil status. Nutrient availability in the soil, especially nitrogen and phosphorus, has been postulated to explain the abrupt transitions between savannas and seasonal forests in tropical regions. Plants from these two biomes may present different nutritional strategies to cope with nitrogen and phosphorus limitation. We used two congeneric pairs of trees — each pair with a species from the savanna and another from the neighboring seasonal forest — to test whether savanna and forest species presented different nutritional strategies during their early development. We cultivated 56 individuals from each of these species in a hydroponics system with four treatments: (1) complete Hoagland solution, (2) Hoagland solution without nitrogen, (3) Hoagland solution without phosphorus, and (4) Hoagland solution without nitrogen and phosphorus. After 45 days, we harvested the plants and measured total biomass, root to shoot ratio, height, leaf area, and specific leaf area. Overall, savanna species were lighter, shorter, with smaller leaves, higher specific leaf areas, and higher root to shoot ratios when compared to the forest species. Nitrogen increased the performance of species from both biomes. Phosphorus improved the performance of the forest species and caused toxicity symptoms in the savanna species. Hence, savanna and forest species presented different demands and were partially distinct already as seedlings concerning their nutritional strategies.

Em regiões tropicais, savanas e florestas estacionais, biomas altamente diversos, ocorrem lado a lado, sob o mesmo clima. Sendo assim, esse mosaico não pode ser explicado somente por variáveis climáticas, devendo ser considerada a frequência e intensidade de incêndios, disponibilidade de água e status do solo. A disponibilidade de nutrientes no solo, especialmente nitrogênio e fósforo, tem sido postulada para explicar as transições abruptas entre savanas e florestas estacionais nos trópicos. Espécies vegetais desses dois biomas podem apresentar estratégias nutricionais diferentes para lidar com a limitação tanto de nitrogênio como de fósforo. Utilizamos dois pares de árvores congenéricas — cada par com uma espécie típica de savana e outra de floresta estacional vizinha — para testar se as espécies da savana e da floresta apresentaram estratégias nutricionais diferentes durante seu desenvolvimento inicial. Cultivamos 56 indivíduos de cada uma dessas espécies em um sistema hidropônico com quatro tratamentos: (1) solução Hoagland completa, (2) solução Hoagland sem nitrogênio, (3) solução Hoagland sem fósforo e (4) solução Hoagland sem nitrogênio e fósforo. Após 45 dias, colhemos as plantas e medimos a biomassa total, a relação raiz / parte aérea, altura, área foliar e área foliar específica. No geral, as espécies savânicas foram mais leves, menores em altura, área foliar e área foliar específica e apresentaram maiores razões entre biomassa radicular por biomassa aérea quando comparadas às espécies florestais. A oferta de nitrogênio aumentou o desempenho das espécies de ambos biomas. O fósforo melhorou o desempenho das espécies florestais e causou sintomas de toxicidade nas espécies savânicas. Concluímos que, já como mudas, espécies congenéricas de savana e floresta apresentaram demandas distintas e foram parcialmente diferentes em relação a suas estratégias nutricionais.

Role of beneficial microbes with nitrogen and phosphorous levels on canola productivity / Papel de microbes benéficos em associação com níveis de nitrogênio e fósforo na produtividade de canola

A research was conducted to evaluate the impact of various nitrogen and phosphorus levels along with beneficial microbes to enhance canola productivity. The research was carried out at Agronomy Research Farm, The University of Agriculture Peshawar in winter 2016-2017. The experiment was conducted in randomized complete block factorial design. The study was comprised of three factors including nitrogen (60, 120 and 180 kg ha-¹), phosphorous (70, 100 and 130 kg ha-¹) and beneficial microbes (with and without BM). A control treatment with no N, P and BM was also kept for comparison. Application of beneficial microbes significantly increased pods plant, seed pod, seed filling duration, 1000 seed weight, biological yield and seed yield as compared to control plots. Nitrogen applied at the rate of 180 kg ha-¹ increased pods plant-¹, seed pod, seed filling duration, seed weight, biological yield and seed yield. Maximum pods plant-¹, seed pod, early seed filling, heavier seed weight, biological yield, seed yield, and harvest index were observed in plots treated with 130 kg.ha-¹ phosphorous. As comparison, the combine treated plots have more pods plant-¹, seeds pod-¹, seed filling duration, heaviest seeds, biological yield, seed yield and harvest index as compared to control plots. It is concluded that application of beneficial microbes with N and P at the rate of 180 kg ha-¹ and 130 kg ha-¹, respectively, increased yield and its attributes for canola.

Uma pesquisa foi realizada para avaliar o impacto de vários níveis de nitrogênio e fósforo, juntamente com micróbios benéficos, para aumentar a produtividade da canola. A pesquisa foi realizada no inverno de 2016-17 no Agronomy Research Farm, Universidade de Agricultura do Peshawar. O experimento foi conduzido por planejamento fatorial aleatorizado em blocos. O estudo focou-se em três fatores, incluindo o teor de nitrogênio, N, (60, 120 e 180 kg.ha-¹), o teor de fósforo, P, (70, 100 e 130 kg ha-¹) e a presença de micróbios benéficos (com BM e sem BM). Para fins de comparação, um tratamento controle sem N, P e BM também foi incluído no estudo. A aplicação de micróbios benéficos aumentou significativamente as vagens das plantas e de sementes, a duração do enchimento das sementes, o peso de 1000 sementes, o rendimento biológico e o rendimento de sementes em comparação com os resultados do controle. O nitrogênio aplicado na taxa de 180 kg ha-¹ aumentou as vagens por planta, vagem, duração do enchimento, peso da semente, rendimento biológico e rendimento de sementes. Vagens máximas por planta, vagem, enchimento precoce de sementes, peso maior de semente, rendimento biológico, rendimento de sementes e índice de colheita foram observados em parcelas tratadas com 130 kg.ha-¹ de fósforo. Em comparação aos blocos cultivados de controle, os blocos cultivados tratados combinados têm mais vagens por planta e sementes por vagem, maior duração do enchimento das sementes, maior número de sementes mais pesadas e maior rendimento biológico, rendimento de sementes e índice de colheita. Conclui-se que a aplicação de micróbios benéficos junto com N e P nas doses de 180 kg ha-¹ e 130 kg ha-¹, respectivamente, aumentou a produtividade e atributos de produtividade para a canola.

Effects of a high-phosphorus diet on the gut microbiota in CKD rats.

OBJECTIVE: To investigate whether high-phosphorus diets alter gut microbiota in healthy rats and chronic kidney disease (CKD) rats. METHODS: In this 4-week randomized controlled trial, healthy rats and CKD rats were fed a regular-phosphorus (Pi: 0.8%) and high-phosphorus (Pi: 1.2%) diet. The subjects were divided into four groups: sham-group rats with regular-phosphorus diet intervention (CTL group), sham-group rats with high-phosphorus diet intervention (CTLP group), CKD model rats with regular-phosphorus diet intervention (CKD group), and CKD model rats with high-phosphorus diet intervention (CKDP group). The V3-V4 region of the 16S rRNA gene was sequenced to study the effect of a high-phosphorus diet on gut microbiota. RESULTS: A high-phosphorus intervention increased systolic blood pressure (SBP) and parathyroid hormone (PTH) in CTL and CKD rats but did not change serum creatinine and 25(OH)D levels. After the high-phosphorus diet, serum phosphate and fibroblast growth factor 23 (FGF23) increased in the CKDP group compared with the CKD group. The gut microbiota was significantly altered after intervention with a high-phosphorus diet in CTL and CKD group rats. A high-phosphorus diet reduced the Shannon index values of gut microbiota in all rats. The Chao1 and Ace indexes were decreased in the CTL group after high-phosphorus diet intervention. Some microbial genera were elevated significantly after high-phosphorus dietary intervention, such as Blautia and Allobaculum. The main bacteria linked to SBP and FGF23 also correlated directly with creatinine. After high-phosphorus diet intervention, the bacteria Prevotella were positively related to SBP in CTLP and CKDP groups. CONCLUSIONS: High-phosphorus diets were associated with adverse changes in gut microbiota and elevated SBP, which may have adverse consequences for long-term health outcomes.

The Effect of Religious Dietary Cultures on Food Nitrogen and Phosphorus Footprints: A Case Study of India.

The excessive consumption of nitrogen (N) and phosphorus (P), two vital nutrients for living organisms, is associated with negative environmental and health impacts. While food production contributes to a large amount of N and P loss to the environment, very little N and P is consumed as food. Food habits are affected by multiple regulations, including the dietary restrictions and dictates of various religions. In this study, religion-sensitive N-Calculator and P-Calculator approaches were used to determine the impact of religious dietary culture on the food N and P footprints of India in the major religious communities. Using 2013 data, the food N footprint of Hindus, Muslims, Christians, and Buddhists was 10.70, 11.45, 11.47, and 7.39 kg-N capita-1 year-1 (10.82 kg-N capita-1 year-1 was the national average), and the food P footprint was 1.46, 1.58, 1.04. and 1.58 kg-P capita-1 year-1 (1.48 kg-P capita-1 year-1 was the national average). The findings highlight the impact of individual choice on the N and P food footprints, and the importance of encouraging the followers of religion to follow a diet consistent with the food culture of that religion. The results of this study are a clear indication of the requirement for religion-sensitive analyses in the collecting of data pertinent to a particular country for use in making government policies designed to improve the recycling of food waste and the treatment of wastewater.

Determination of elemental distribution and evaluation of elemental concentration in single Saccharomyces cerevisiae cells using single cell-inductively coupled plasma mass spectrometry.

Single-cell analysis using inductively coupled plasma mass spectrometry (SC-ICP-MS) is a method to obtain qualitative and quantitative information of the elemental content and distribution of single cells. Six intrinsic target elements were analyzed in yeast cells at different cell growth phases cultured in medium with different phosphorus concentrations (0, 7, 14 mM) to study its effect on cell growth and composition. SC-ICP-MS results were compared with those obtained by the acid digestion and the average ratio was 0.81. The limits of detection of this method were 0.08, 2.54, 12.5, 0.02, 0.02, and 0.08 fg cell-1 for Mg, P, K, Mn, Cu, and Zn, respectively. During the exponential growth phase, the cells exhibited higher elemental contents, wider distribution for most elements, and larger cell size in comparison to the stationary growth phase. Phosphorus-free conditions reduced the average P content in single cells of stationary growth phase from 650 to 80 fg. Phosphorus deficiency led to decreasing intracellular concentrations not only of P but also of K and Cu, and to increasing Zn concentration after 48 h. Mg maintained its concentration at ∼0.11 fg µm-3 and did not change significantly under the three investigated conditions after 48 h. Accordingly, Mg content was successfully used to estimate the intracellular concentration of other intrinsic elements in single yeast cells. SC-ICP-MS is suited to determine target elements in single yeast cells, and allows the study of heterogeneity of cell composition and effects of stressors on the elemental content, distribution, and concentrations of intrinsic elements.

Low nitrogen availability inhibits the phosphorus starvation response in maize (Zea mays ssp. mays L.).

BACKGROUND: Nitrogen (N) and phosphorus (P) are macronutrients essential for crop growth and productivity. In cultivated fields, N and P levels are rarely sufficient, contributing to the gap between realized and potential production. Fertilizer application increases nutrient availability, but is not available to all farmers, nor are current rates of application sustainable or environmentally desirable. Transcriptomic studies of cereal crops have revealed dramatic responses to either low N or low P single stress treatments. In the field, however, levels of both N and P may be suboptimal. The interaction between N and P starvation responses remains to be fully characterized. RESULTS: We characterized growth and root and leaf transcriptomes of young maize plants under nutrient replete, low N, low P or combined low NP conditions. We identified 1555 genes to respond to our nutrient treatments, in one or both tissues. A large group of genes, including many classical P starvation response genes, were regulated antagonistically between low N and P conditions. An additional experiment over a range of N availability indicated that a mild reduction in N levels was sufficient to repress the low P induction of P starvation genes. Although expression of P transporter genes was repressed under low N or low NP, we confirmed earlier reports of P hyper accumulation under N limitation. CONCLUSIONS: Transcriptional responses to low N or P were distinct, with few genes responding in a similar way to the two single stress treatments. In combined NP stress, the low N response dominated, and the P starvation response was largely suppressed. A mild reduction in N availability was sufficient to repress the induction of P starvation associated genes. We conclude that activation of the transcriptional response to P starvation in maize is contingent on N availability.

Update on Calcium and Phosphorus Requirements of Preterm Infants and Recommendations for Enteral Mineral Intake.

BACKGROUND: With current Ca and P recommendations for enteral nutrition, preterm infants, especially VLBW, fail to achieve a bone mineral content (BMC) equivalent to term infants. During the first 3 years, most notably in light at term equivalent age (<-2 Z score) VLBW infants' BMC does not catch up. In adults born preterm with VLBW or SGA, lower adult bone mass, lower peak bone mass, and higher frequency of osteopenia/osteoporosis have been found, implying an increased risk for future bone fractures. The aim of the present narrative review was to provide recommendation for enteral mineral intake for improving bone mineral accretion. METHODS: Current preterm infant mineral recommendations together with fetal and preterm infant physiology of mineral accretion were reviewed to provide recommendations for improving bone mineral accretion. RESULTS: Current Ca and P recommendations systematically underestimate the needs, especially for Ca. CONCLUSION: Higher enteral fortifier/formula mineral content or individual supplementation is required. Higher general mineral intake (especially Ca) will most likely improve bone mineralization in preterm infants and possibly the long-term bone health. However, the nephrocalcinosis risk may increase in infants with high Ca absorption. Therefore, individual additional enteral Ca and/or P supplementations are recommended to improve current fortifier/formula mineral intake.

Mass spectrometry-based quantification and spatial localization of small organic acid exudates in plant roots under phosphorus deficiency and aluminum toxicity.

Low-molecular-weight organic acid (OA) extrusion by plant roots is critical for plant nutrition, tolerance to cations toxicity, and plant-microbe interactions. Therefore, methodologies for the rapid and precise quantification of OAs are necessary to be incorporated in the analysis of roots and their exudates. The spatial location of root exudates is also important to understand the molecular mechanisms directing OA production and release into the rhizosphere. Here, we report the development of two complementary methodologies for OA determination, which were employed to evaluate the effect of inorganic ortho-phosphate (Pi) deficiency and aluminum toxicity on OA excretion by Arabidopsis roots. OA exudation by roots is considered a core response to different types of abiotic stress and for the interaction of roots with soil microbes, and for decades has been a target trait to produce plant varieties with increased capacities of Pi uptake and Al tolerance. Using targeted ultra-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS), we achieved the quantification of six OAs in root exudates at sub-micromolar detection limits with an analysis time of less than 5 min per sample. We also employed targeted (MS/MS) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) to detect the spatial location of citric and malic acid with high specificity in roots and exudates. Using these methods, we studied OA exudation in response to Al toxicity and Pi deficiency in Arabidopsis seedlings overexpressing genes involved in OA excretion. Finally, we show the transferability of the MALDI-MSI method by analyzing OA excretion in Marchantia polymorpha gemmalings subjected to Pi deficiency.

Cerebral phosphoester signals measured by 31P magnetic resonance spectroscopy at 3 and 7 Tesla.

Abnormal cell membrane metabolism is associated with many neuropsychiatric disorders. Free phosphomonoesters and phosphodiesters, which can be detected by in vivo 31P magnetic resonance spectroscopy (MRS), are important cell membrane building blocks. However, the quantification of phosphoesters has been highly controversial even in healthy individuals due to overlapping signals from macromolecule membrane phospholipids (MP). In this study, high signal-to-noise ratio (SNR) cerebral 31P MRS spectra were acquired from healthy volunteers at both 3 and 7 Tesla. Our results indicated that, with minimal spectral interference from MP, the [phosphocreatine (PCr)]/[phosphocholine (PC) + glycerophosphocholine (GPC)] ratio measured at 7 Tesla agreed with its value expected from biochemical constraints. In contrast, the 3 Tesla [PCr]/[PC+GPC] ratio obtained using standard spectral fitting procedures was markedly smaller than the 7 Tesla ratio and than the expected value. The analysis suggests that the commonly used spectral model for MP may fail to capture its complex spectral features at 3 Tesla, and that additional prior knowledge is necessary to reliably quantify the phosphoester signals at low magnetic field strengths when spectral overlapping is significant.

The "Fat but Fit" Paradigm and Bone Health in Young Adults: A Cluster Analysis.

The fat but fit paradox has suggested that obese individuals with good fitness levels have lower cardiometabolic risk compared to individuals with normal weight but lower fitness levels. This paradigm has not been explored in the context of bone health. The aim of this study was to test whether categories of fat but fit paradigm assessed by body fat percentage and handgrip strength holds up in young adults and to analyze the relationship between fat but fit categories and bone outcomes. Cluster cross-sectional analyses of data from 499 young adults aged 18 to 30 from Toledo and Cuenca, Spain were conducted. Body fat percentage, handgrip strength, bone mineral content (BMC), bone mineral density (BMD), and dietary nutrients such as, proteins, magnesium, calcium, phosphorus, potassium, and vitamin D were assessed. Cluster analysis of body fat percentage and handgrip z scores resulted in a classification of four clusters that could be interpreted according to Fat Unfit (FU), Unfat Unfit (UU), Fat Fit (FF) and Unfat Fit (UF) categories. ANCOVA models showed that young adults in clusters with higher handgrip strength levels (FF, UF) and with higher key bone nutrients levels (UF) had significantly higher total BMC values than their peers in the UU and FU cluster categories, after controlling for sex, age and height. This study provides two novel conclusions in relation to the fat but fit paradigm: first, it confirms the construct of the four clusters of body fat percentage and handgrip strength, and second, it reinforces the predictive validity of the fat but fit paradigm categories, indicating the positive effect, although it may not just be a causal relationship, of muscular strength and key bone nutrients on counteracting the negative effect of obesity on bone health.

Associations between intake of calcium, magnesium and phosphorus and risk of pancreatic cancer: a population-based, case-control study in Minnesota.

Experimental studies suggest that abnormal levels of Ca, Mg and phosphorus are implicated in pancreatic carcinogenesis. We investigated the associations between intakes of these minerals and the risk of pancreatic cancer in a case-control study conducted in 1994-1998. Cases of pancreatic cancer (n 150) were recruited from all hospitals in the metropolitan area of the Twin Cities and Mayo Clinic, Minnesota. Controls (n 459) were randomly selected from the general population and frequency matched to cases by age, sex and race. All dietary variables were adjusted for energy intake using the residual method prior to data analysis. Logistic regression was performed to evaluate the associations between intake of three nutrients examined and the risk of pancreatic cancer. Total intake of Ca (936 v. 1026 mg/d) and dietary intake of Mg (315 v. 331 mg/d) and phosphorus (1350 v. 1402 mg/d) were significantly lower in cases than in controls. After adjustment for confounders, there were not significant associations of total and dietary intakes of Ca, Mg and phosphorus with the risk of pancreatic cancer. In addition, no significant interactions exist between intakes of these minerals and total fat on pancreatic cancer risk. In conclusion, the present study does not suggest that intakes of Ca, Mg and phosphorus were significantly associated with the risk of pancreatic cancer.

A Stable Quaternized Chitosan-Black Phosphorus Nanocomposite for Synergetic Disinfection of Antibiotic-Resistant Pathogens.

Antibiotics are widely used for treatment of bacterial infections, and their overuse has contributed to microbial resistance. Currently, an alternative antibiotic-free therapy for inactivating bacteria is of great interest. Black phosphorus (BP), a biocompatible and nontoxic rising-star two-dimensional layered material, has gained remarkable interest in many bioapplications including biosensing, cancer therapy, drug delivery, and also antibacterial treatment. However, BP nanosheets suffer from instability in ambient environments due to rapid oxidation and degradation. To address this issue, BP nanosheets were modified with quaternized chitosan (QCS) by electrostatic adsorption to prepare a BP-QCS composite for photothermal/pharmaco treatment of bacterial infection. The BP-QCS has obviously enhanced solubility and chemical stability in aqueous suspensions. We have demonstrated that under near-infrared (NIR) irradiation, the BP-QCS can synergistically inactivate more than 95% methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) and Escherichia coli within 10 min with a dose of only 75 µg/mL in vitro. Meanwhile, the BP-QCS composite under NIR can synergistically inactivate 98% S. aureus in vivo. Furthermore, the BP-QCS suspensions at effective antibacterial concentrations have negligible cytotoxicity and in vivo toxicity.

Agronomic efficiency and profitability of cotton on integrated use of phosphorus and plant microbes / Eficiência e rentabilidade agronômica do algodão no uso integrado de fósforo e micróbios vegetais

Cotton crop, plays a significant role in Pakistan's economy by ruling a prominent place in edible oil and local textile industry. Phosphorus (P) inaccessibility and deficiency of soil organic matter are the key restraints for low crop productivity in cotton. Therefore, a two years field study was designed during 2014-15, to explore the influence of phosphate solubilizing bacteria (PSB), farmyard manure (FYM), poultry manure (PM) and inanimate sources of P on various physiological, growth, yield and quality parameters of cotton crop at CCRI Multan. Field responses of seeds inoculated with two distinctive phosphate solubilizing bacteria (PSB) strains viz. S0 = control, S1 =strain-1, S2 = strain-2 and eight organic, inorganic P sources viz., P0= control, P1 = 80 kg ha-1 P from inorganic source, P2 = 80 kg ha-1 P from FYM, P3 = 80 kg ha-1 P from PM, P4 = 40 kg ha-1 P from FYM + 40 kg ha-1 P from inorganic source, P5 = 40 kg ha-1 P from PM + 40 kg ha-1 P from inorganic source, P6 = 80 kg ha-1 P from FYM + 40 kg ha-1 P from inorganic source, P7 = 80 kg ha-1 P from PM + 40 kg ha-1 P from inorganic source and P8 = 40 kg ha-1 P from FYM + 40 kg ha-1 P from PM were evaluated. Results revealed that inoculation of seeds with PSB and collective use of inorganic and organic sources of P had considerably increased the yield contributing attributes in cotton. However, the treatment P7 (80 kg P ha-1 from PM + 40 kg P ha-1 from inorganic source) in coincidence with seeds inoculated with PSB (S1) produced taller plant, maximum boll weight, significantly higher LAI and CGR. Significantly higher seed cotton yield, lint yield, fiber length and maximum BCR of 1.95 and 1.81 was also obtained from the P7 treatment during both crop-growing seasons. In conclusion, combined use of 80 kg P ha-1 from PM + 40 kg P ha-1 from inorganic source and cotton seeds inoculated with strain-1 improved phosphorus uptake ensuing in greater consumption of photo-assimilates for maximum growth and yield.

A safra de algodão, desempenha um papel significativo na economia do Paquistão, ao ocupar um lugar de destaque no óleo comestível e na indústria têxtil local. A inacessibilidade e a deficiência de fósforo (P) da matéria orgânica do solo são as principais restrições para a baixa produtividade das culturas em algodão. Portanto, um estudo de campo de dois anos foi desenvolvido durante 2014-15, para explorar a influência de bactérias solubilizantes de fosfato (PSB), esterco de capoeira (FYM), esterco de aves (PM) e fontes inanimadas de P sobre vários fatores fisiológicos, crescimento, rendimento e parâmetros de qualidade da cultura do algodão no CCRI Multan. Respostas de campo de sementes inoculadas com duas cepas distintas de bactérias solubilizantes de fosfato (PSB) viz. S0 = controle, S1 = cepa-1, S2 = cepa-2 e oito fontes orgânicas de P inorgânicas viz., P0 = controle, P1 = 80 kg ha-1 P de fonte inorgânica, P2 = 80 kg ha-1 P de FYM, P3 = 80 kg ha-1 P de PM, P4 = 40 kg ha-1 P de FYM + 40 kg ha-1 P de fonte inorgânica, P5 = 40 kg ha-1 P de PM + 40 kg ha-1 P de fonte inorgânica, P6 = 80 kg ha-1 P de FYM + 40 kg ha-1 P de fonte inorgânica, P7 = 80 kg ha-1 P de PM + 40 kg ha-1 P de fonte inorgânica e P8 = 40 Foram avaliados kg ha-1 P da FYM + 40 kg ha-1 P da PM. Os resultados revelaram que a inoculação de sementes com PSB e o uso coletivo de fontes inorgânicas e orgânicas de P aumentaram consideravelmente os atributos que contribuem para a produtividade no algodão. No entanto, o tratamento P7 (80 kg P ha-1 da PM + 40 kg P ha-1 da fonte inorgânica) em coincidência com sementes inoculadas com PSB (S1) produziu planta mais alta, peso máximo de cápsula, IAF e CGR significativamente maiores. Produtividade significativamente maior do algodão, rendimento de fiapos, comprimento da fibra e BCR máximo de 1,95 e 1,81 também foi obtida a partir do tratamento P7 durante as duas épocas de cultivo. Em conclusão, o uso combinado de 80 kg P ha-1 de PM + 40 kg P ha-1 de fonte inorgânica e sementes de algodão inoculadas com a cepa-1 melhoraram a captação de fósforo, resultando em maior consumo de foto assimilados para obter crescimento e produtividade máximos.

Delivery of Cas13a/crRNA by self-degradable black phosphorus nanosheets to specifically inhibit Mcl-1 for breast cancer therapy.

Mcl-1 amplification has been observed in breast cancer and demonstrated as a key determinant of breast cancer cell survival. However, the clinical use of available effective Mcl-1-specific inhibitors for breast cancer treatment remains a challenge. An RNA-guided CRISPR/Cas13a system targeting RNAs can be used to specifically knock down mRNA expression in mammalian cells. The goal of this work is to develop a self-degradable nanoplatform based on polylysine (PLL)-functionalized black phosphorus (PBP) for the delivery of Cas13a/crRNA complexes to specifically inhibit Mcl-1 at transcriptional level for breast cancer therapy. The constructed Cas13a/crRNA complex is delivered into the cytoplasm by PBP via endocytosis, followed by endosomal escape based on the biodegradation of PBP, and this efficiently knocks down the specific gene at transcriptional level up to an efficiency of 58.64%. Through designing CRISPR RNA crMcl-1, Mcl-1 can be specifically knocked down at transcriptional level in breast cancer cells, resulting in the down-regulation of the expression of Mcl-1 protein and inhibition of the cell activity. Notably, PBP/Cas13a/crMcl-1 shows an excellent tumor suppression efficacy up to 65.16% after intratumoral injection. Therefore, biodegradable PBP is an ideal nanoplatform for the delivery of CRISPR/Cas13a, which could provide a potential strategy for gene therapy.

Black phosphorus quantum dots encapsulated in anionic waterborne polyurethane nanoparticles for enhancing stability and reactive oxygen species generation for cancer PDT/PTT therapy.

Black phosphorus quantum dots (BPQDs) with excellent biocompatibility, outstanding photothermal and photodynamic efficacies have attracted significant attention in cancer therapy. However, the low environmental stability and poor dispersity of BPQDs limit their practical applications. In the present work, biocompatible anionic waterborne polyurethane (WPU) nanoparticles were synthesized from castor oil to encapsulate the BPQDs. The WPU-BPQDs with a BPQDs loading capacity of about 13.8% (w/w) exhibited significantly improved dispersion and environmental stability without affecting the photothermal efficiency of BPQDs. Intriguingly, it was found that WPU encapsulation led to significant enhancement in the reactive oxygen species (ROS) generation of BPQDs, which indicated the enhanced photodynamic efficacy of the encapsulated BPQDs as compared to the bare BPQDs. The effect of solution pH on the ROS generation efficiency of BPQDs and the pH variation caused by BPQDs degradation was then investigated to explore the possible mechanism. In acidic solution, ROS generation was suppressed, while BPQDs degradation led to the acidification of the solution. Fortunately, after being encapsulated inside the WPU nanoparticles, the degradation rate of BPQDs became slower, while the acidic environment around BPQDs was favorably regulated by WPU nanoparticles having a special electrochemical double layer consisting of interior COO- and exterior NH(Et3)+, thus endowing the WPU-BPQDs-boosted production of ROS as compared to the bare BPQDs. Considering the undesired acidic tumor environment, this unique pH regulation effect of WPU-BPQDs would be beneficial for in vivo photodynamic efficacy. Both in vitro and in vivo experiments showed that WPU-BPQDs could effectively improve photodynamic therapy (PDT) and maintain outstanding photothermal therapy (PTT) effects. Together with the excellent dispersity, biocompatibility, and easy biodegradability, WPU-BPQDs can be a promising agent for PDT/PTT cancer treatments.

In situ sprayed NIR-responsive, analgesic black phosphorus-based gel for diabetic ulcer treatment.

The treatment of diabetic ulcer (DU) remains a major clinical challenge due to the complex wound-healing milieu that features chronic wounds, impaired angiogenesis, persistent pain, bacterial infection, and exacerbated inflammation. A strategy that effectively targets all these issues has proven elusive. Herein, we use a smart black phosphorus (BP)-based gel with the characteristics of rapid formation and near-infrared light (NIR) responsiveness to address these problems. The in situ sprayed BP-based gel could act as 1) a temporary, biomimetic "skin" to temporarily shield the tissue from the external environment and accelerate chronic wound healing by promoting the proliferation of endothelial cells, vascularization, and angiogenesis and 2) a drug "reservoir" to store therapeutic BP and pain-relieving lidocaine hydrochloride (Lid). Within several minutes of NIR laser irradiation, the BP-based gel generates local heat to accelerate microcirculatory blood flow, mediate the release of loaded Lid for "on-demand" pain relief, eliminate bacteria, and reduce inflammation. Therefore, our study not only introduces a concept of in situ sprayed, NIR-responsive pain relief gel targeting the challenging wound-healing milieu in diabetes but also provides a proof-of-concept application of BP-based materials in DU treatment.