Synthesis, structure and reactivity of the novel tetratin species bromoheptakis(2,6-diethylphenyl)-cyclotetrastannabutane

Metallation of ArBr (Ar = 2,6-diethylphenyl) with Li powder in diethyl ether, followed by addition of stannous chloride at low temperature does not give the expected oligomeric diarylstannane but an essentially quantitative yield of the novel tetrastannabutane [{SnAr2}3SnArBr]. Some reactions of the new species are reported.

Computer vision and color measurement techniques for inline monitoring of cheese curd syneresis

Optical characteristics of stirred curd were simultaneously monitored during syneresis in a 10-L cheese vat using computer vision and colorimetric measurements. Curd syneresis kinetic conditions were varied using 2 levels of milk pH (6.0 and 6.5) and 2 agitation speeds (12.1 and 27.2 rpm). Measured optical parameters were compared with gravimetric measurements of syneresis, taken simultaneously. The results showed that computer vision and colorimeter measurements have potential for monitoring syneresis. The 2 different phases, curd and whey, were distinguished by means of color differences. As syneresis progressed, the backscattered light became increasingly yellow in hue for circa 20 min for the higher stirring speed and circa 30 min for the lower stirring speed. Syneresis-related gravimetric measurements of importance to cheese making (e.g., curd moisture content, total solids in whey, and yield of whey) correlated significantly with computer vision and colorimetric measurements..

Effect of milk fat concentration and gel firmness on syneresis during curd stirring in cheese-making

An experiment was undertaken to investigate the effect of milk fat level (0%, 2.5% and 5.0% w/w) and gel firmness level at cutting (5, 35 and 65 Pa) on indices of syneresis, while curd was undergoing stirring. The curd moisture content, yield of whey, fat in whey and casein fines in whey were measured at fixed intervals between 5 and 75 min after cutting the gel. The casein level in milk and clotting conditions was kept constant in all trials. The trials were carried out using recombined whole milk in an 11 L cheese vat. The fat level in milk had a large negative effect on the yield of whey. A clear effect of gel firmness on casein fines was observed. The best overall prediction, in terms of coefficient of determination, was for curd moisture content using milk fat concentration, time after gel cutting and set-to-cut time (R2 = 0.95).

Influence of curd cutting programme and stirring speed on the prediction of syneresis indices in cheese-making using NIR light backscatter

An NIR reflectance sensor, with a large field of view and a fibre-optic connection to a spectrometer for measuring light backscatter at 980 nm, was used to monitor the syneresis process online during cheese-making with the goal of predicting syneresis indices (curd moisture content, yield of whey and fat losses to whey) over a range of curd cutting programmes and stirring speeds. A series of trials were carried out in an 11 L cheese vat using recombined whole milk. A factorial experimental design consisting of three curd stirring speeds and three cutting programmes, was undertaken. Milk was coagulated under constant conditions and the casein gel was cut when the elastic modulus reached 35 Pa. Among the syneresis indices investigated, the most accurate and most parsimonious multivariate model developed was for predicting yield of whey involving three terms, namely light backscatter, milk fat content and cutting intensity (R2 = 0.83, SEy = 6.13 g/100 g), while the best simple model also predicted this syneresis index using the light backscatter alone (R2 = 0.80, SEy = 6.53 g/100 g). In this model the main predictor was the light backscatter response from the NIR light back scatter sensor. The sensor also predicted curd moisture with a similar accuracy.

Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were ∼1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime (τ2 = 1.9–3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there is significant nuclear absorption of flavanols. This advanced imaging using two-photon excitation and biophysical techniques described here will prove valuable for probing the intracellular trafficking and functions of flavanols, such as EGCG, which is the major flavanol of green tea.

The socioeconomics of food crop production and climate change vulnerability: a global scale quantitative analysis of how grain crops are sensitive to drought

Many studies warn that climate change may undermine global food security. Much work on this topic focuses on modelling crop-weather interactions but these models do not generally account for the ways in which socio-economic factors influence how harvests are affected by weather. To address this gap, this paper uses a quantitative harvest vulnerability index based on annual soil moisture and grain production data as the dependent variables in a Linear Mixed Effects model with national scale socio-economic data as independent variables for the period 1990-2005. Results show that rice, wheat and maize production in middle income countries were especially vulnerable to droughts. By contrast, harvests in countries with higher investments in agriculture (e.g higher amounts of fertilizer use) were less vulnerable to drought. In terms of differences between the world's major grain crops, factors that made rice and wheat crops vulnerable to drought were quite consistent, whilst those of maize crops varied considerably depending on the type of region. This is likely due to the fact that maize is produced under very different conditions worldwide. One recommendation for reducing drought vulnerability risks is coordinated development and adaptation policies, including institutional support that enables farmers to take proactive action.

Variation in the global-scale impacts of climate change on crop productivity due to climate model uncertainty and adaptation

Crop production is inherently sensitive to fluctuations in weather and climate and is expected to be impacted by climate change. To understand how this impact may vary across the globe many studies have been conducted to determine the change in yield of several crops to expected changes in climate. Changes in climate are typically derived from a single to no more than a few General Circulation Models (GCMs). This study examines the uncertainty introduced to a crop impact assessment when 14 GCMs are used to determine future climate. The General Large Area Model for annual crops (GLAM) was applied over a global domain to simulate the productivity of soybean and spring wheat under baseline climate conditions and under climate conditions consistent with the 2050s under the A1B SRES emissions scenario as simulated by 14 GCMs. Baseline yield simulations were evaluated against global country-level yield statistics to determine the model's ability to capture observed variability in production. The impact of climate change varied between crops, regions, and by GCM. The spread in yield projections due to GCM varied between no change and a reduction of 50%. Without adaptation yield response was linearly related to the magnitude of local temperature change. Therefore, impacts were greatest for countries at northernmost latitudes where warming is predicted to be greatest. However, these countries also exhibited the greatest potential for adaptation to offset yield losses by shifting the crop growing season to a cooler part of the year and/or switching crop variety to take advantage of an extended growing season. The relative magnitude of impacts as simulated by each GCM was not consistent across countries and between crops. It is important, therefore, for crop impact assessments to fully account for GCM uncertainty in estimating future climates and to be explicit about assumptions regarding adaptation.

Climate change impacts on crop productivity in Africa and South Asia

Climate change is a serious threat to crop productivity in regions that are already food insecure. We assessed the projected impacts of climate change on the yield of eight major crops in Africa and South Asia using a systematic review and meta-analysis of data in 52 original publications from an initial screen of 1144 studies. Here we show that the projected mean change in yield of all crops is − 8% by the 2050s in both regions. Across Africa, mean yield changes of − 17% (wheat), − 5% (maize), − 15% (sorghum) and − 10% (millet) and across South Asia of − 16% (maize) and − 11% (sorghum) were estimated. No mean change in yield was detected for rice. The limited number of studies identified for cassava, sugarcane and yams precluded any opportunity to conduct a meta-analysis for these crops. Variation about the projected mean yield change for all crops was smaller in studies that used an ensemble of > 3 climate (GCM) models. Conversely, complex simulation studies that used biophysical crop models showed the greatest variation in mean yield changes. Evidence of crop yield impact in Africa and South Asia is robust for wheat, maize, sorghum and millet, and either inconclusive, absent or contradictory for rice, cassava and sugarcane.

Diversity of cultivars and other plant resources used at habitation sites in the Llanos de Mojos, Beni, Bolivia: Evidence from macrobotanical remains, starch grains, and phytoliths

Although sparsely populated today, the Llanos de Mojos, Bolivia, sustained large sedentary societies in the Late Holocene (ca. 500 to 1400 AD). In order to gain insight into the subsistence of these people, we undertook macrobotanical and phytolith analyses of sediment samples, and starch grain and phytolith analyses of artifact residues, from four large habitation sites within this region. Macrobotanical remains show the presence of maize (Zea mays), squash (Cucurbita sp.), peanut (Arachis hypogaea), cotton (Gossypium sp.), and palm fruits (Arecaceae). Microbotanical results confirm the widespread use of maize at all sites, along with manioc (Manihot esculenta), squash, and yam (Dioscorea sp.). These integrated results present the first comprehensive archaeobotanical evidence of the diversity of plants cultivated, processed, and consumed, by the pre-Hispanic inhabitants of the Amazonian lowlands of Bolivia.

Pre-Columbian landscape impact and agriculture in the Monumental Mound region of the Llanos de Moxos, lowland Bolivia

We present a multiproxy study of land use by a pre-Columbian earth mounds culture in the Bolivian Amazon. The Monumental Mounds Region (MMR) is an archaeological sub-region characterized by hundreds of pre-Columbian habitation mounds associated with a complex network of canals and causeways, and situated in the forest–savanna mosaic of the Llanos de Moxos. Pollen, phytolith, and charcoal analyses were performed on a sediment core from a large lake (14 km2), Laguna San José (14°56.97′S, 64°29.70′W).We found evidence of high levels of anthropogenic burning from AD 400 to AD 1280, corroborating dated occupation layers in two nearby excavated habitation mounds. The charcoal decline pre-dates the arrival of Europeans by at least 100 yr, and challenges the notion that the mounds culture declined because of European colonization. We show that the surrounding savanna soils were sufficiently fertile to support crops, and the presence of maize throughout the record shows that the area was continuously cultivated despite land-use change at the end of the earthmounds culture. We suggest that burning was largely confined to the savannas, rather than forests, and that pre-Columbian deforestation was localized to the vicinity of individual habitation mounds, whereas the inter-mound areas remained largely forested.

An optimized method for the extraction of bacterial mRNA from plant roots infected with Escherichia coli O157:H7

Analysis of microbial gene expression during host colonization provides valuable information on the nature of interaction, beneficial or pathogenic, and the adaptive processes involved. Isolation of bacterial mRNA for in planta analysis can be challenging where host nucleic acid may dominate the preparation, or inhibitory compounds affect downstream analysis, e.g., quantitative reverse transcriptase PCR (qPCR), microarray, or RNA-seq. The goal of this work was to optimize the isolation of bacterial mRNA of food-borne pathogens from living plants. Reported methods for recovery of phytopathogen-infected plant material, using hot phenol extraction and high concentration of bacterial inoculation or large amounts of infected tissues, were found to be inappropriate for plant roots inoculated with Escherichia coli O157:H7. The bacterial RNA yields were too low and increased plant material resulted in a dominance of plant RNA in the sample. To improve the yield of bacterial RNA and reduce the number of plants required, an optimized method was developed which combines bead beating with directed bacterial lysis using SDS and lysozyme. Inhibitory plant compounds, such as phenolics and polysaccharides, were counteracted with the addition of high-molecular-weight polyethylene glycol and hexadecyltrimethyl ammonium bromide. The new method increased the total yield of bacterial mRNA substantially and allowed assessment of gene expression by qPCR. This method can be applied to other bacterial species associated with plant roots, and also in the wider context of food safety.

High pressure intensification of cassava resistant starch (RS3) yields

Cassava starch, typically, has resistant starch type 3 (RS3) content of 2.4%. This paper shows that the RS3 yields can be substantially enhanced by debranching cassava starch using pullulanase followed by high pressure or cyclic high-pressure annealing. RS3 yield of 41.3% was obtained when annealing was carried out at 400 MPa/60°C for 15 min, whereas it took nearly 8 h to obtain the same yield under conventional atmospheric annealing at 60°C. The yield of RS3 could be further significantly increased by annealing under 400MPa/60°C pressure for 15 min followed by resting at atmospheric pressure for 3 h 45 min, and repeating this cycle for up to six times. Microstructural surface analysis of the product under a scanning electron microscope showed an increasingly rigid density of the crystalline structure formed, confirming higher RS3 content.

Effects of water availability on free amino acids, sugars, and acrylamide-forming potential in potato

Irrigation is used frequently in potato cultivation to maximize yield, but water availability may also affect the composition of the crop, with implications for processing properties and food safety. Five varieties of potatoes, including drought-tolerant and -sensitive types, which had been grown with and without irrigation, were analyzed to show the effect of water supply on concentrations of free asparagine, other free amino acids, and sugars and on the acrylamide-forming potential of the tubers. Two varieties were also analyzed under more severe drought stress in a glasshouse. Water availability had profound effects on tuber free amino acid and sugar concentrations, and it was concluded that potato farmers should irrigate only if necessary to maintain the health and yield of the crop, because irrigation may increase the acrylamide-forming potential of potatoes. Even mild drought stress caused significant changes in composition, but these differed from those caused by more extreme drought stress. Free proline concentration, for example, increased in the field-grown potatoes of one variety from 7.02 mmol/kg with irrigation to 104.58 mmol/kg without irrigation, whereas free asparagine concentration was not affected significantly in the field but almost doubled from 132.03 to 242.26 mmol/kg in response to more severe drought stress in the glasshouse. Furthermore, the different genotypes were affected in dissimilar fashion by the same treatment, indicating that there is no single, unifying potato tuber drought stress response.

Pre-Columbian land use in the ring-ditch region of the Bolivian Amazon

The nature and extent of pre-Columbian (pre-1492 AD) human impact in Amazonia is a contentious issue. The Bolivian Amazon has yielded some of the most impressive evidence for large and complex pre-Columbian societies in the Amazon basin, yet there remains relatively little data concerning the land use of these societies over time. Palaeoecology, when integrated with archaeological data, has the potential to fill these gaps in our knowledge. We present a 6,000-year record of anthropogenic burning, agriculture and vegetation change, from an oxbow lake located adjacent to a pre-Columbian ring-ditch in north-east Bolivia (13°15’44” S, 63°42’37” W). Human occupation around the lake site is inferred from pollen and phytoliths of maize (Zea mays L.) and macroscopic charcoal evidence of anthropogenic burning. First occupation around the lake was radiocarbon dated to ~2500 years BP. The persistence of maize in the record from ~1850 BP suggests that it was an important crop grown in the ringditch region in pre-Columbian times, and abundant macroscopic charcoal suggests that pre-Columbian land management entailed more extensive burning of the landscape than the slash-and-burn agriculture practised around the site today. The site was occupied continuously until near-modern times, although there is evidence for a decline in agricultural intensity or change in land use strategy, and possible population decline, from ~600-500 BP. The long and continuous occupation, which predates the establishment of rainforest in the region, suggests that pre-Columbian land use may have had a significant influence on ecosystem development at this site over the last ~2000 years.

Targeted activation of toll-like receptors: conjugation of a toll-like receptor 7 agonist to a monoclonal antibody maintains antigen binding and specificity

Therapeutic activation of Toll-like receptors (TLR) has potential for cancer immunotherapy, for augmenting the activity of anti-tumor monoclonal antibodies (mAbs), and for improved vaccine adjuvants. A previous attempt to specifically target TLR agonists to dendritic cells (DC) using mAbs failed because conjugation led to non-specific binding and mAbs lost specificity. We demonstrate here for the first time the successful conjugation of a small molecule TLR7 agonist to an anti-tumour mAb (the anti-hCD 20 rituximab) without compromising antigen specificity. The TLR7 agonist UC-1V150 was conjugated to rituximab using two conjugation methods and yield, molecular substitution ratio, retention of TLR7 activity and specificity of antigen binding were compared. Both conjugation methods produced rituximab-UC-1V150 conjugates with UC-1V150 : rituximab ratio ranging from 1:1 to 3:1 with drug loading quantified by UV spectroscopy and drug substitution ratio verified by MALDI TOF mass spectroscopy. The yield of purified conjugates varied with conjugation method, and dropped as low as 31% using a method previously described for conjugating UC-1V150 to proteins, where a bifunctional crosslinker was firstly reacted with rituximab, and secondly to the TLR7 agonist. We therefore developed a direct conjugation method by producing an amine-reactive UV active version of UC-1V150, termed NHS:UC-1V150. Direct conjugation with NHS:UC-1V150 was quick and simple and gave improved conjugate yields of 65-78%. Rituximab-UC-1V150 conjugates had the expected pro-inflammatory activity in vitro (EC50 28-53 nM) with a significantly increased activity over unconjugated UC-1V150 (EC50 547 nM). Antigen binding and specificity of the rituxuimab-UC-1V150 conjugates was retained, and after incubation with human peripheral blood leukocytes, all conjugates bound strongly only to CD20-expressing B cells whilst no non-specific binding to CD20-negative cells was observed. Selective targeting of Toll-like receptor activation directly within tumors or to DC is now feasible.