Powder diffraction from protein powders using in-house diffractometers is an effective tool for identification and monitoring of protein crystal forms and artifacts. As an alternative to conventional powder diffractometers a single crystal diffractometer equipped with an X-ray micro-source can be used to collect powder patterns from 1 µl samples. Using a small-angle X-ray scattering (SAXS) camera it is possible to collect data within minutes. A streamlined program has been developed for the calculation of powder patterns from pdb-coordinates, and includes correction for bulk-solvent. A number of such calculated powder patterns from insulin and lysozyme have been included in the powder diffraction database and successfully used for search-match identification. However, the fit could be much improved if peak asymmetry and multiple bulk-solvent corrections were included. When including a large number of protein data sets in the database some problems can be foreseen due to the large number of overlapping peaks in the low-angle region, and small differences in unit cell parameters between pdb-data and powder data. It is suggested that protein entries are supplied with more searchable keywords as protein name, protein type, molecular weight, source organism etc. in order to limit possible hits.

Aqueous solutions of reconstituted whey protein- concentrate (WPC) & isolate (WPI) powders were sonicated at 20 kHz in a batch process for 1–60 min. Sonication at 20 kHz increased the clarity of WPC solutions largely due to the reduction in the size of the suspended insoluble aggregates. The gel strength of these solutions when heated at 80°C for 20 min also increased with sonication, while gelation time and gel syneresis were reduced. These improvements in gel strength were observed across a range of initial pH values, suggesting that the mechanism for gel promotion is different from the well known effects of pH. Examining the microstructure of the whey protein gels indicated a compact network of densely packed whey protein aggregates arising from ultrasound treatment. Comparable changes were not observed with whey protein isolate solutions, which may reflect the absence of larger aggregates in the initial solution or differences in composition.

A method which was developed for assaying the extracellular lipases of psychrotrophic bacteria in milk (Stead, 1983, 1984) and which uses the fluorogenic substrate 4-methylumbelliferyl oleate has been adapted for use with skim milk powder (SMP), whey powder (WP) and whey protein concentrate (WPC). A five-fold increase in the concentration of sodium taurocholate (NaTC), in the mixture of NaTC and cetyltrimethylammonium bromide needed to dissociate lipase from milk proteins, removed the excessive sensitivity of the assay to variations in the concentrations of SMP, WP or WPC incorporated. Commercially available pancreatic lipase provided a suitable standard of lipolytic activity and as little as 1–2 μ could be detected in each assay system.

The effect of preheat treatment, evaporation and drying in a commercial plant on the denaturation of βlactoglobulin and α-lactalbumin, their incorporation into the casein micelle and the heat stability characteristics of the milks and powders were determined. Preheat treatments between 110 °C for 2 min and 120 °C for 3 min denatured between 80 and 91% of β-lactoglobulin and between 33 and 45% of α-lactalbumin. Evaporation increased the extent of denaturation but spray drying did not increase it further. The incorporation of α-lactalbumin and βlactoglobulin into the micelles was markedly less than the amount that denatured and was not a constant ratio to it. Heat coagulation times at 140 °C of milks, concentrates and powders diluted to the original milk concentration were measured as a function of pH. In general, the greater the collective heat treatment, the shorter the time required to achieve coagulation. Spray drying shifted the peak positions in the pH-heat coagulation time profiles. In contrast, heat coagulation times (measured at 120 °C) of concentrates and powders diluted to 20% total solids content increased with the severity of the preheat treatment. Surprisingly, spray drying markedly increased the heat coagulation times of the diluted concentrates.

The solubility of milk protein concentrate (MPC) powders was influenced by the method used for preparing the concentrate, drying conditions, and the type of dryer used. Increasing total solids of the ultrafiltered concentrates (23% total solids, TS) by diafiltration to 25% TS or evaporation to 31% TS decreased the solubility of MPC powders (80–83% protein, w/w dry basis), with ultrafiltration followed by evaporation to higher total solids having the greater detrimental effect on solubility. High shear treatment (homogenisation at 350/100 bar, microfluidisation at 800 bar or ultrasonication at 24 kHz, 600 watts) of ultrafiltered and diafiltered milk protein concentrates prior to spray drying increased the nitrogen solubility of MPC powders (82% protein, w/w dry basis). Of the treatments applied, microfluidisation was the most effective for increasing nitrogen solubility of MPC powders after manufacture and during storage. Manufacture of MPC powders (91% protein, w/w dry basis) prepared on two different pilot-scale dryers (single stage or two stage) from milk protein concentrates (20% TS) resulted in powders with different nitrogen solubility and an altered response to the effects of microfluidisation. Microfluidisation (400, 800 and 1200 bar) of the concentrate prior to drying resulted in increased long term solubility of MPC powders that were prepared on a single stage dryer but not those produced on a two stage spray dryer. This work demonstrates that microfluidisation can be used as a physical intervention for improving MPC powder solubility. Interactions between the method of preparation and treatment of concentrate prior to drying, the drying conditions and dryer type all influence MPC solubility characteristics.

With either Na caseinate or skim-milk powder, 1% (w/v) additions of pepsin, chymotrypsin or Alcalase and an incubation time of 24 h at 37°C gave the best hydrolysates for subsequent plastein production. In spite of the fact that peptide species which were produced by treatment with the 3 enzymes were entirely different, they were all capable of producing qualitatively similar amounts of plastein when concentrated and further incubated with proteinase, suggesting that the identity of peptide components was relatively unimportant. αs1-, β- and κ- caseins, Na-caseinate and skim-milk powder all led to plastein products with broadly similar properties, confirming the comparative unimportance of peptide composition. This also indicated that for most practical applications fractionation of initial protein mixtures (as in many food products) would not be justified. Optimum peptide concentration for plastein formation with pepsin and caseinate hydrolysates was 20–35% (w/v) with an optimum pH range of 4–5; the best peptide molecular weight varied between 400 and 800 and the optimum temperature was either 37°C for 24 h or 50°C for 4–6 h. Higher temperatures (70°C) gave more rapid plastein formation but poorer yields. Lower temperatures (e.g. 20°C) gave similar yields, but incubation times required to be extended to at least 48 h. The same proteinase should be used for plastein synthesis as that used in the initial hydrolysis stage, or else further hydrolysis (over hydrolysis) could occur during the plastein formation stage causing lower yields due to differences in the specificity of the 2 enzymes. Even under ideal conditions plastein yields never exceeded 27% of the weight of peptide taken and at the present time it seems unlikely that the plastein reaction will have anything more than curiosity value. Economics must preclude its application in low-cost, high volume situations such as generally exist within the food industry.

In the previous paper(1) it was shown that the heat treatment to which a milk powder is subjected during manufacture may result in the production of two distinct types of protein insolubility, namely insolubility due to denaturation of the protein while still in solution, and insolubility caused by over-heating the milk powder while in a dry state. The insolubility due to denaturation was shown to be independent of the temperature of reconstitution of the milk powder. On the other hand the insolubility due to dry heating was found to vary with the temperature of reconstitution, the protein being relatively insoluble when reconstituted in water held at 20° C, but completely soluble when mechanically stirred for 30 min. in water held at 90° C. It was suggested that this latter type of insolubility might be due to the removal of water of imbibition from the protein particles, since such an explanation would account for the effect of hot water in restoring the solubility of the protein. In the previous paper, however, observations on the effect of temperature of reconstitution were limited to the two extreme temperatures, i.e. 20 and 90° C.

No reliable information is at present available with regard to the effect of process of manufacture on the solubility of the protein of milk powders. Hunziker(1), it is true, gives a very well-balanced discussion of the subject, but it is apparent from his summary of the views of other workers that these are seldom based on adequate experimental evidence. In fact the only points on which there is general agreement among workers in this field are that the proteins of spray-dried powders are, on the whole, extremely soluble, while those of roller-dried powders are relatively insoluble. Moreover, even where experimental data are available, there remain many unexplained points. For example, Hunziker shows that the solubility of the proteins of a milk powder may vary according to the temperature at which the powder is reconstituted—a higher temperature giving a greater solubility. Such an observation cannot at present be brought into line with current theories regarding the properties of denatured proteins.

The effects of high pressure (HP) treatment (100–400 MPa at 10–60°C) on the solubility of milk protein concentrate (MPC) powders were tested. The solubility, measured at 20°C, of fresh MPC powders made with no HP treatment was 66%. It decreased by 10% when stored for 6 weeks at ambient temperature (∼20°C) and continued to decrease to less than 50% of its initial solubility after 12 months of storage. Of the combinations of pressure and heat used, a pressure of 200 MPa at 40°C applied to the concentrate before spray drying was found to be the most beneficial for improved solubility of MPC powders. This combination of pressure/heat improved the initial cold water solubility to 85%. The solubility was maintained at this level after 6 weeks storage at ambient temperature and 85% of the initial solubility was preserved after 12 months. The improved solubility of MPC powders on manufacture and on storage are attributed to an altered surface composition arising from an increased concentration of non-micellar casein in the milk due to HP treatment prior to drying. The improved solubility of high protein powders (95% protein) made from blends of sodium caseinate and whey protein isolate compared with MPC powders (∼85% protein) made from ultrafiltered/diafiltered milk confirmed the detrimental role of micellar casein on solubility. The results suggest that increasing the non-micellar casein content by HP treatment of milk or use of blends of sodium caseinate and whey proteins are strategies that may be used to obtain high protein milk powders with enhanced solubility.

We tested the hypothesis that milk proteins, through microencapsulation, guarantee protection against bioactive substances in coffee silverskin extracts. Therefore, the aim of this study was to carry out technological, nutritional and physicochemical characterisation of a coffee silverskin extract microencapsulated using instant skim milk powder and whey protein concentrate as wall materials. The aqueous extract of coffee silverskin was spray-dried using 10% (w/v) skim milk powder and whey protein concentrate. The samples were characterised by determining the water content, water activity, particle size distribution, colour analysis and total phenolic compound content as well as antioxidant activity using 2,2-diphenyl-radical 1-picrylhydrazyl scavenging methods, nitric oxide radical inhibition and morphological analysis. The product showed water activity within a range that ensured greater stability, and the reduced degradation of the dried coffee silverskin extract with whey protein concentrate resulted in better rehydration ability. The luminosity parameter was higher and the browning index was lower for the encapsulated samples than for the pure coffee silverskin extract. The phenolic compound content (29.23 ± 8.39 and 34.00 ± 8.38 mg gallic acid equivalents/g for the coffee silverskin extract using skimmed milk powder and whey protein concentrate, respectively) and the antioxidant activity of the new product confirmed its potential as a natural source of antioxidant phenolic compounds. We conclude that the dairy matrices associated with spray drying preserved the bioactive and antioxidant activities of coffee silverskin extracts.

Amateur and professional athletes often consume protein supplements to accelerate muscle gain; however, it has been suggested that these products not only are associated with risks when consumed excessively. Several recent reports have indicated that certain products are contaminated with heavy metals. Therefore, in this study, we aimed to investigate protein powders in Hungary for heavy metal contamination. A total of 22 commercially available protein powders (including whey, vegan, and beef based) were purchased on the internet for testing. We analysed the samples using laser-induced breakdown spectroscopy (LIBS) and inductively coupled plasma mass spectrometry (ICP-MS) to assess heavy metal contamination. The products were analysed for the presence of 16 elements (Be, Al, Cr, Mn, Co, Ni, Cu, As, Se, Rb, Cd, Sb, Cs, Ba, Hg, and Pb). The LIBS spectral analysis revealed the characteristics of the protein elements (C, C2, H, N, and O) and alkaline metals (Ca, Na, K, and Mg), which were consistent with the previous results. Neither LIBS nor ICP-MS measurements detected significant heavy metal content in the investigated samples above the limit specified in the regulations. Heavy metal contamination of protein supplements can be a serious health threat. Based on the varied results of the previous studies, it is prudent to include testing for heavy metals as part of the routine and mandatory quality control of these products.

Dietary interventions are a common practice in the poultry industry to promote optimal performance and health of animals. Here, we aim at assessing the influence of supplementing broiler diets with dry whey powder (DWP) and whey protein concentrate (WPC) on nutrient coefficient of apparent ileal digestibility (CAID) and productive performance. Cecal microbiota composition was also determined using Illumina amplicon sequencing. Dietary treatments were control diet (no supplementation of DWP or WPC), 60-DWP (60 g/kg of DWP), and 80-WPC (80 g/kg of WPC). One-day-old male broilers were randomly assigned to one of three treatments, and housed in floor pens. In Trial 1, 90 1-day-old chicks were allocated to three pens/treatment, with 10 birds/pen, during 21 days for CAID evaluation. Diet 60-DWP increased Ca CAID (P=0.041), while diet 80-WPC improved Ca and P CAID (P<0.001 and 0.002, respectively) when compared with control diet. In Trial 2, 810 one-day-old chicks were allocated to nine pens/treatment, with 30 birds/pen, during 42 days. Feeding chickens with 60-DWP and 80-WPC increased their BW, average daily gain (ADG) and feed intake (FI) during the starter (P<0.001 for all variables) and grower-finisher periods (P<0.001 for BW and FI, and P=0.048 for ADG), and during the entire feeding period (P<0.05), when compared with control diet. Diets 60-DWP and 80-WPC reduced the feed conversion ratio of chickens during the starter period (P<0.001 and 0.003, respectively), while 60-DWP reduced this parameter during the entire feeding period (P=0.048), when compared to control diet. At day 42, cecal microbial communities of chickens that were fed with 60-DWP and 80-WPC differed from those fed with control diet (R=0.776, P=0.008; and R=0.740, P=0.008, respectively). The abundance of Bacteroides fragilis, Bacteroides spp., Escherichia coli/Shigella flexneri and Megamonas furniformis increased when 60-DWP and 80-WPC diets were offered, while the presence of Helicobacter pullorum decreased. Lactobacillus salivarius consistently increased in chickens with better feed conversion ratio, which were those fed with 60-DWP. The results obtained in the present study indicate that growth of chickens is improved by DWP and WPC supplementation because of a higher mineral digestibility, increased feed intake and modulation of cecal microbiota communities.

Feeding monogastric livestock in organic agriculture is challenging due to several tradeoffs between animal welfare aspects, resource efficiency, as well as ecological and social sustainability. Organic standards may even increase such conflicts, as is currently the case with upcoming new regulations regarding restrictions of feed sources for organic pigs in Europe. In order to contribute data for balancing reasons to minimize tradeoffs, we compared four different piglet diets, each targeted to reach a high protein quality by either a high proportion of soybean cake (SOY), inclusion of milk powder (MILK), fermentatively produced lysine (LYS) or conventional potato protein (POT). All diets were designed to meet the nutritional requirements of piglets in the best possible way, however they all represented different conflicts with either organic regulations or sustainability goals. In each of five consecutive runs, respectively three litters were assigned to every dietary treatment, resulting in 15 litters per treatment in total. In each litter, seven focus animals were defined. The piglets were studied from birth until 58 days of age. They were weaned at day 46 and sold from the farm at day 58. Piglets were individually weighed at an average age of 3, 21, 43, 50 and 58 days with simultaneous assessment of body condition score (BCS) and prevalence of diarrhea. Feed intake (FI) was recorded litter wise weekly, starting from week three. Feed conversion ratio (FCR) was calculated for the period after weaning. Statistical analysis was executed using linear mixed effect models. Regarding FI, FCR and daily weight gains, no treatment effect was found. Only at day 21, BCS was lower for piglets receiving POT. Prevalence of diarrhea increased after weaning for all treatments. All four tested diets led to similar weight gains and feed conversion in the piglets. Animals fed diet POT recovered better from diarrhea compared to the other treatments. A high soybean cake content or lysine supplementation in the diet was disadvantageous with regard to the occurrence of diarrhea. LYS diet led to signs of threonine deficit, indicating that lysine addition alone may not solve the issue. The addition of milk powder provided no extra benefit. In recognition of the health benefits, the use of 5% potato protein, even if it is sourced from conventional production, must still be considered as a sustainable option for feeding organic piglets. The sustainability implications are discussed in the paper.

The purpose of the present study was to test the anti-inflammatory and blood glucose (BG)-regulating capacity of strawberries in a mouse model of diet-induced obesity. A total of thirty-six male C57BL/6J mice were randomly divided into four groups (nine mice per group). Mice were fed a low-fat diet (LF, 13 % fat), the LF supplemented with 2·6 % freeze-dried strawberry powder (LFSB), a high-fat diet (HF, 44 % fat) or the HF supplemented with 2·6 % strawberry powder (HFSB). Blood samples were collected to measure BG, inflammation and systemic markers for endocrine function of pancreas and adipose tissue. Splenocytes were harvested at the end of the study and activated with either anti-cluster of differentiation (CD) 3/anti-CD28 antibodies or lipopolysaccharide to test immune responsiveness. The HF increased non-fasted BG, insulin, soluble intracellular adhesion molecule-1, E-selectin, leptin, resistin and plasminogen activator protein-1 (P < 0·05). High dietary fat decreased IL-4 production from activated splenocytes (P < 0·05). BG concentrations were lower in the mice supplemented with SB (10·64 mmol/l) compared to the non-supplemented mice (11·37 mmol/l; P = 0·0022). BG values were approximately 6·5 % lower in the supplemented mice. Additionally, SB lowered plasma C-reactive protein in the LFSB group compared to the other three groups (P < 0·05). The dietary intake of SB approximated one human serving of strawberries. These results, although modest, support a promising role for dietary strawberries in reducing the risks associated with obesity and diabetes, and regulating the levels of inflammatory markers in non-obese individuals.

Thirty-six pigs were weaned at 21 days of age and were given diets supplemented with either dicalcium phosphate (DCP) or defluorinated rock phosphate (DFP) from 4 weeks of age until slaughter at 50 kg live weight. From 4 to 9 weeks of age the diets contained either skim milk powder (SKIM) or soya bean meal (SOY) as the main protein supplement. The dietary treatments were arranged in a 2 × 2 factorial design. From 9 weeks of age until slaughter, SOY was the only protein supplement used. Four 7-day measurements of phosphorus, calcium and nitrogen balance were made on each pig. Faecal endogenous P was estimated in two pigs per treatment by the 32P dilution method.

There were no significant effects of P supplement on performance or on apparent absorption, true absorption or retention of P. The apparent absorption and retention of Ca were both 0·07 lower with DFP than with DCP in the period from 9 weeks of age to slaughter (P< 0·001).

From 4 to 9 weeks of age there was no effect of protein supplement on performance. From 9 weeks of age to slaughter, when all pigs were given SOY, those which had received SKIM in the previous period grew slightly faster (P<0·05); their growth-rate overall, from 4 weeks of age to slaughter, was also significantly better (P <0·05).

Pigs given SKIM showed a higher apparent absorption of P (P < 0·001) and higher apparent absorption (P<0·01) and retention (P<0·001) of both Ca and N from 4 to 9 weeks of age. Again, there were carry-over effects in the period from 9 weeks of age to slaughter, although the trends observed in the first period were reversed; pigs previously given SKIM showed significantly lower apparent absorption and retention of both P and Ca, and apparent absorption of N, than those previously given SOY.

There were no significant effects of dietary treatments on the ash, Ca or P content of the fourth metacarpal.

Even before his accession King James is said to have given a verbal promise to Thomas Percy, the future conspirator, that the Catholics would be granted some degree of liberty. There is, in the very nature of things, no written evidence of this promise, and the King later denied having given it, but the Catholics firmly believed that some assurance of toleration had been given. But by the beginning of 1604 all hope of toleration for papists was gone. On 19 February, James I protested ‘his utter detestation of their superstitious religion, and that he was so far from favouring it, as if he thought his son and heir after him would give any toleration thereunto, he would wish him fairly buried before his eyes’. And fairly buried he was in 1612.

On 22 February, 1604, a proclamation was issued ordering all Jesuits and seminary priests to depart the kingdom before 19 March. On the same day the fine of £20 a month for recusancy was again put in force, and was made to include the whole period since the King’s coming, thus negativing what little relief had been granted.

On 24 April, a bill was introduced in the lower house, classing Catholics with forgers, perjurers and outlaws, and disabling them from sitting in parliament, while an ‘Act for the due execution of the statutes against Jesuits, seminary priests and recusants’ made in this session, not only reenforced all the laws made in Elizabeth’s reign but even added to their severity. On the third reading of this bill Viscount Montague courageously denounced it, and the following day found himself in the Fleet for his ‘scandalous and offensive speech’. Further proclamations followed and on 16 July the bloody persecution broke out again when John Sugar, a priest, and Robert Grissold his servant, were executed at Warwick.