Monitoring the crystallization of amylose–lipid complexes during maize starch melting by synchrotron x‐ray diffraction

Most starch granules exhibit a natural crystallinity, with different diffraction patterns according to their botanical origin: A‐type from cereals and B‐type from tubers. The V polymorph results essentially from the complexing of amylose with compounds such as iodine, alcohols, or lipids. The intensity and nature of phase transitions (annealing, melting, polymorphic transitions, recrystallization, etc.) induced by hydrothermal treatments in crystalline structures are related to temperature and water content. Despite its small concentration, the lipid phase present mainly in cereal starches has a large influence on starch properties, particularly in complexing amylose. The formation of Vh crystalline structures was observed by synchrotron x‐ray diffraction in native maize starch heated at intermediate and high moisture contents (between 19 and 80%). For the first time, the crystallization of amylose–lipid complexes was evidenced in situ by x‐ray diffraction without any preliminary cooling, at heating rates corresponding to the usual conditions for differential scanning calorimetry experiments. For higher water contents, the crystallization of Vh complexes clearly occurred at 110–115°C. For intermediate water contents, mixed A + Vh (or B + Vh for high amylose starch) diffraction diagrams were recorded. Two mechanisms can be involved in amylose complexing: the first relating to crystallization of the amylose and lipid released during starch gelatinization, and the second to crystalline packing of separate complexed amylose chains (amorphous complexes) present in native cereal starches. © 1999 John Wiley & Sons, Inc. Biopoly 50: 99–110, 1999     

Effect of F-2 and T-2 fusariotoxins on experimental Cryptosporidium baileyi infection in chickens

The course of Cryptosporidium baileyi infection in chickens fed with different doses of fusariotoxins was compared with that of control groups. F-2 toxin levels of 0.187–1.5 mg kg⁻¹ and T-2 toxin levels of 0.187–6.0 mg kg⁻¹ were investigated. The experimental amimals were orally infected with 6 × 10⁵ C. baileyi oocysts at 1 week of age. Total daily oocyst output was monitored by a quantitative method. Acquired immunity was tested at the age of 4 weeks, by ELISA and by a challenge infection with an equal number of oocysts, upon recovery from the primary infection. The results show that in chickens kept on the lower doses of F-2 and T-2 toxins, the parasite infection ran a similar course to that in the control groups, and the animals became resistant to re-infection. However, when higher doses (2.0–6.0 mg kg⁻¹) of T-2 toxin were used, a depression of weight gain was observed with some other physiological parameters (PCV, weight of bursa, weight of thymus, skin thickness in PHA-P skin test) also indicating toxic effect and, simultaneously, the oocyst output decreased significantly and the patent period was slightly prolonged. Although certain modifications of the immune response could be revealed, the chickens became resistant to re-infection. Only early (1 week of age) parasite infection and 6 mg kg⁻¹ T-2 toxin in the feed significantly depressed body weight gain and immunity.     

Changes on the structure, consistency, physicochemical and viscoelastic properties of corn (Zea mays sp.) under different nixtamalization conditions

Aqueous corn (Zea mays sp.) dispersions with 1% (w/w) of calcium hydroxide/weight of corn were cooked at 90 °C (nixtamalization) for different times (10, 30, 50, 70 and 90 min), making five treatments. A control lot (nine treatments), was cooked for 10, 30, 50, 60, 70, 90, 120, 150 and 180 min. Then, processed corn samples were washed and milled to obtain masa (dough). Scanning electron microscopy, calorimetry, viscoamilography and rheological analysis were used to characterize the corn samples. The corn micrographs showed that the nixtamalization modified the shape of the starch granules and the protein bodies. Starch granules from nixtamalized samples, were round shaped, while control samples, showed polygonal shape. Proteins from nixtamalized samples usually exhibited two transition endotherms, while in control samples, only one transition was seen, suggesting some relationship with gelatinized starch. The nixtamalization shortened the corn cooking time to develop a proper texture in masa to obtain good quality tortillas, as seen in their consistencies. All corn masa samples showed weak gel-like viscoelastic behavior with the elastic modulus (G′) higher than the loss modulus (G″), over all strain and frequency domains.     

Synthesis, crystalline structure and photoluminescence investigations of the new trivalent rare earth complexes (Sm, Eu and Tb) containing 2-thiophenecarboxylate as sensitizer

New complexes with the general formula [RE(TPC)₃ · (H₂O)₂], where RE=Eu³⁺, Sm³⁺, Gd³⁺, Tb³⁺ and TPC=2-thiophenecarboxylate, have been prepared and investigated by photoluminescence spectroscopy. These compounds were characterized by complexometric titration, elemental analyses and infrared spectroscopy. The X-ray crystal structure has been determined for the [Eu(TPC)₃ · (H₂O)₂] compound, indicating that this complex is in dimeric form bridged by two carboxylate ions with monoclinic crystal system and space group P2₁/n. The coordination polyhedron can be described as a distorted square antiprism, where six oxygen atoms belong to the TPC ligand and two oxygen atoms belong to the water molecules, with site symmetry close to C_2v. The theoretical value of the intensity parameter , which is in agreement with the experimental one, indicates that the Eu³⁺ ion is in a highly polarizable chemical environment. Based on the luminescence spectra, the energy transfer from the ligand triplet state (T) of TPC to the excited levels of the Eu³⁺ ion is discussed. The emission quantum efficiency of the ⁵D₀ emitting level of the Eu³⁺ ion was also determined. In the case of the Tb³⁺ ion, the photoluminescence data show the high emission intensity of the characteristic transitions ⁵D₄ → ⁷F_J (J=0-6), indicating that the TPC ligand is a good sensitizer. It is also noticed that the complexes with the Eu³⁺ and Tb³⁺ ions are more luminescent than the complex with the Sm³⁺ ion.     

Variability of the EMG mean power frequency: A study on the trapezius muscle

Calculation of the EMG mean power frequency (MPF) is a common procedure applied in evaluation of the frequency shift associated with local muscle fatigue. Variations of the MPF that are unrelated to muscle fatigue may jeopardize the estimation of the frequency shift. Different kinds of variation include random variation and systematic variation due to changes in posture or load. In a previous article we have evaluated the systematic linear variation of the MPF. The aim of the present study was to examine the random variation. Data sequences of 10 s, each obtained from nonfatigued trapezius muscle of 19 healthy subjects, were examined over a functional range of load and joint angles with multiple regression analysis. The random variation was evaluated with residual analysis. The residual standard deviation within the whole group was 10% for surface recordings and 13% for intramuscular recordings. If only within-subject variation was considered, the corresponding values were 5 and 8%. Based on this, confidence and prediction intervals for the regression models were calculated. Ninety-five percent confidence intervals were ±1–3% around the regression surfaces, whereas 95% prediction intervals for single measurements were as large as ±20–26% for the whole group, and ±11–20% if only within-subject variations were considered. Assessment of localized muscle fatigue using single MPF estimates should therefore be avoided. Multiple measurements and regression analysis are discussed as methods to minimize the effects of random variations.     

Amylose content and chemical modification effects on the extrusion of thermoplastic starch from maize

The effects of starch structural properties and starch modification on extruder operation were monitored via die pressure, motor torque, mean residence time and specific mechanical energy (SME). The structural properties studied involved variations in the ratios of amylose and amylopectin as well as the effect of a hydroxypropylated starch on the fore mentioned extruder properties. A full factorial design of experiments (DOE) was used to then determine the influence of starch type (unmodified starches with 0%, 28%, 50% and 80% amylose; 80% amylose hydroxypropylated starch) and screw speed (250, 300 and 350 rpm) on these processing parameters. The effects of starch type and screw speed on extrusion operation that were systematically investigated using the DOE and have provided valuable insight into the relationships between starch structure and processing. The design of experiments showed that starch type for both unmodified and modified maize had a statistically significant effect on parameters such as torque, die pressure and specific mechanical energy and that screw speed also significantly effected specific mechanical energy. Residence time distributions differed according to starch type (amylose content, hydroxypropylation) and screw speed. The additional study of residence time distribution also gave an indication of the degree of mixing in the extruder. Starch type variations were apparent at low screw speed however at higher screw speed the influence of starch type decreased significantly.     

The effect of season on the properties of wild boar (Sus scrofa L.) semen

The objective of the study was to determine the properties of wild boar semen and their changes in annual cycle. During a 14-month study period, 167 ejaculates were sampled from 3 mature boars. In each ejaculate the volume of liquid fraction, percentage of spermatozoa motility, spermatozoa concentration and the total number of spermatozoa were determined. The activity of acid and alkaline phosphatase, and aspartate aminotransferase in the fresh semen plasma was also measured. It was shown that wild boar ejaculates did not differ from those of domestic boars, and the semen of the highest volume, concentration and number of spermatozoa was produced in late autumn. The spermatozoa motility was the lowest in summer. The activity of aspartate aminotransferase and alkaline phosphatase in the semen plasma increased with shortening of the light period.     

Observations on the history of Dutch physical stature from the late-Middle Ages to the present

In the late-Middle Ages and at the onset of the early modern period, the Dutch population was taller than in the first half of the 19th century. This inference is partially based on skeletal evidence, mainly collected by the Dutch physical anthropologist George Maat and his co-workers. A spectacular increase in Dutch heights began in the second half of the 19th century and accelerated in the second half of the 20th century. At the end of the 20th century, the Dutch became tallest in the world.     

Structural analysis of DNA-PKcs: modelling of the repeat units and insights into the detailed molecular architecture

DNA-dependent protein kinase (DNA-PK) is part of the eukaryotic DNA double strand break repair pathway and as such is crucial for maintenance of genomic stability, as well as for V(D)J (variable-diversity-joining) recombination. The catalytic subunit of DNA-PK (DNA-PKcs) belongs to the phosphatidylinositol-3 (PI-3) kinase-like kinase (PIKK) superfamily and is comprised of approximately 4100 amino acids. We have used a novel repeat detection method to analyse this enormous protein and have identified two different types of helical repeat motifs in the N-terminal region of the sequence, as well as other previously unreported features in this repeat region. A comparison with the ATMs, ATRs, and TORs show that the features identified are likely to be conserved throughout the PIKK superfamily. Homology modelling of parts of the DNA-PKcs sequence has been undertaken and we have been able to fit the models to previously obtained electron microscopy data. This work provides an insight into the overall architecture of the DNA-PKcs protein and identifies regions of interest for further experimental studies.     

Effect of polymeric cosolutes on calcium pectinate gelation. Part 2. Dextrans and inulin

In a continuation of the work reported in the preceding paper, the effect of chicory root inulin (M_r≈4.5 kD) and branched dextrans of M_r 67, 464 and 2000 kD on gelation of low methoxy pectin (DE 31; 2.0 wt%; pH≈2.9–3.0) on cooling from 90 to 5 °C in the presence of stoichiometric Ca²⁺ has been characterised by low amplitude oscillatory measurements of G′ and G″. As found for other polymeric cosolutes studied previously (oxidised starch, potato maltodextrin, guar gum and locust bean gum), increasing concentrations of dextran or inulin caused a progressive increase of G′ and G″ in the solution state at 90 °C (attributed to segregative interactions promoting formation of calcium-mediated ‘egg-box’ junctions between pectin chains) and a progressive reduction in final moduli at 5 °C (attributed to excessive association of calcium pectinate into large aggregated bundles), with cooling curves crossing those for calcium pectinate alone at 55 °C. For the dextran samples, the ability to promote initial association of pectin chains at high temperature decreased with increasing molecular weight (i.e. with the smallest molecules having the greatest effect, as was also found for guar gum), but the reduction in final modulus on completion of cooling to 5 °C became more pronounced. By the criterion of the decrease in the change in log G′ during cooling, the three dextran samples studied seem roughly comparable in their overall effectiveness in promoting self-association of pectin, with inulin having a substantially smaller effect. Both materials, however, are less effective than the partially depolymerised starches and galactomannans studied previously. It is tentatively suggested that this may be associated with the flexibility introduced by the presence of three single bonds in the predominant glycosidic linkages of both dextran and inulin.     

Effect of polymeric cosolutes on calcium pectinate gelation. Part 3. Gum arabic and overview

Addition of gum arabic (average M_r≈450 kDa; 0.5–2.0 wt%) to solutions of low methoxy pectin (DE 31; 2.0 wt%; pH≈2.9–3.0) with stoichiometric Ca²⁺ caused massive increases in G′ and G″ in the pre-gel state at 90 °C (attributed to segregative interactions promoting formation of calcium-mediated ‘egg-box’ junctions between pectin chains) but had little effect on the gels formed on cooling to 5 °C. This is in marked contrast to the behaviour of other polymeric cosolutes studied in the investigations reported in the two preceding papers, which caused large reductions in gel moduli (attributed to excessive association of calcium pectinate into large aggregated bundles); the difference is tentatively ascribed to strengthening of the calcium pectinate network by divalent counterions to the uronate residues in gum arabic. When the complication of cation exchange was eliminated by extensive dialysis of gum arabic against 100 mM Na⁺ and use of the final dialysate in preparation of mixtures with calcium pectinate, massive increases in G′ and G″ at high temperature were again observed, but with accompanying reductions in moduli at low temperature, which, at gum arabic concentrations above 1.0 wt%, arose from collapse of the developing calcium pectinate network during cooling. The tentative conclusion from this work, and from the two preceding papers, is that enthalpically unfavourable (segregative) interactions between low methoxy pectin and polymeric cosolutes can be relieved in two ways: (i) Ca²⁺-mediated self-association of pectin into compact ordered assemblies which occupy less of the total volume, and (ii) conformational rearrangement of the cosolute molecules to minimise segmental interactions with pectin; conformational rearrangement is inhibited by chain stiffness and by branching; thus polymeric cosolute molecules of limited flexibility are more effective in promoting self-association of pectin than more flexible molecules of comparable size, and branched molecules are more effective than linear chains of comparable stiffness.     

Effect of the insecticidal Galanthus nivalis agglutinin on metabolism and the activities of brush border enzymes in the rat small intestine

With increasing use of lectin genes in crop plants to improve insect resistance, the dietary exposure of humans to lectins will rise and it is necessary to assess whether the presently most favored insecticidal lectin, Galanthus nivalis agglutinin, would be harmful for mammals. Effects of Galanthus nivalis agglutinin on gut and brush border enzymes were studied in rats over a 10-day dietary exposure and compared with those of a known antinutrient, phytohaemagglutinin. At a level that provides insecticidal protection for plants but did not reduce the growth of young rats, Galanthus nivalis agglutinin had negligible effects on the weight and length of the small intestine even though there was a slight, but significant hypertrophy of this tissue. However, the activities of brush border enzymes were affected; sucrase-isomaltase activity was nearly halved and those of alkaline phosphatase and aminopeptidase were significantly increased. Although most of the changes in gut metabolism caused by the incorporation of Galanthus nivalis agglutinin in the diet were less extensive than those found with toxic phytohaemagglutinin, some of them may be potentially deleterious. Thus, further and longer animal studies are needed to establish whether it is safe to use Galanthus nivalis agglutinin in transgenic plants destined for human consumption.     

The influence of the type of contraction on the masseter muscle EMG power spectrum

Different behaviours of the EMG power spectrum across increasing force levels have been reported for the masseter muscle. A factor that could explain these different behaviours may be the type of contraction used, as was recently shown for certain upper limb muscles⁵. The purpose of this study was to compare, between two types of isometric contractions, the behaviour of EMG power spectrum statistics (median frequency (MF) and mean power frequency (MPF)) obtained across increasing force levels. Ten women exerted, while biting in the intercuspal position, three 5 s ramp contractions that increased linearly from 0 to 100% of the maximal voluntary contraction (MVC). They also completed three step contractions (constant EMG amplitude) at each of the following levels: 20, 40, 60 and 80% MVC. EMG signals from the masseter muscle were recorded with miniature surface electrodes. The RMS, as well as the MPF and MF of the power spectrum were calculated at 20, 40, 60 and 80% MVC for each type of contraction. As expected, the RMS values showed similar increases with increasing levels of effort for both types of contractions. Different behaviours for both MPF (contraction^*force interaction, ANOVA, P<0.05) and MF (contraction^*force interaction, ANOVA, P>0.05) across increasing levels of effort were found between the two types of contraction. The use of step contractions gave rise to a decrease of both MPF and MF with increasing force, while the use of ramp contractions gave rise to an increase in both statistics up to at least 40% MVC followed by a decrease at higher force levels. These findings suggest that the type of contraction used does influence the behaviour of the spectral statistics across increasing force levels and that this could explain the differences obtained in previous studies for the masseter muscle.     

Two genetic variants of the crustacean hyperglycemic hormone (CHH) from the Australian crayfish, Cherax destructor: detection of chiral isoforms due to posttranslational modification

From sinus glands of the Australian crayfish Cherax destructor, two genetic variants of the crustacean hyperglycemic hormone (CHH) were isolated by HPLC and fully characterized by mass spectrometry and Edman sequencing. Both CHH A (8350.38 Da) and CHH B (8370.34 Da) consist of 72 amino acid residues, with pyroGlu as N-terminus and an amidated (Val-NH2) C-terminus. They differ in 14 residues (81% identity). Both sequences are significantly different from those of the hitherto known three CHHs of Astacoidea species (Northern hemisphere crayfish), which among themselves are extremely conserved. This may reflect the long, separate evolution of the Astacoidea lineage and the Parastacoidea (Southern hemisphere crayfish) lineage, to which Cherax belongs. CHH A and CHH B genes are expressed at comparable levels, as indicated by the similar amounts of mature peptides in the sinus gland. In addition to each of the major peptides, which share the identical N-terminal tripeptide pyroGlu-Val-L-Phe, one chiral isoform containing pyroGlu-Val-D-Phe was identified. Compared to the main peptides, the amounts of the D-isoforms are lower, but significant, amounting to 30-40% of L-isoforms. These results demonstrate that two genes can give rise to a total of four different peptides in the secretory terminals of the sinus gland. All peptides gave a highly significant hyperglycemic in vivo response in C. destructor.     

Partial characterisation of high-molecular weight glycoconjugates in the trail mucus of the freshwater pond snail Lymnaea stagnalis

We have studied the glycoconjugates in trail mucus of the pond snail Lymnaea stagnalis. The mucus was dissolved with 6 M guanidinium hydrochloride (GuHCl) and the major component was comprised of very high-M_r glycoconjugates that were eluted in the void volume of a Sepharose CL-4B gel-filtration column. This high-M_r material was pooled and thereafter subjected to density gradient centrifugation first in 4 M GuHCl/CsCl and subsequently 0.2 M GuHCl/CsCl to further remove non-glycosylated proteins and DNA. The harvested glycoconjugate pool chromatographed in the void volume of Sepharose CL-2B. However, reduction of disulfide bonds lowered the molecular size of approximately 80% of the void material yielding a major fragment and some minor smaller fragments in gel chromatography. The reduced glycoconjugates were digested with papain and yielded high molecular weight, proteinase-resistant glycopeptides. This fragmentation pattern is similar to that found for oligomeric gel-forming mucins in mammals and the amino acid composition (60% Ser/Thr) and sugar analysis of the glycopeptides is consistent with mucin-like molecules, there being no significant amounts of xylose or uronic acids. The residual 20% of the preparation, which apparently resisted reduction and protease digestion, had a similar amino acid composition to the bulk, but was somewhat different in sugar composition, containing some xylose and a significant amount of glucuronic acid. The two groups of molecules had very different morphologies in the electron microscope. Taken together, these data suggest that trail mucus is a complex mixture of at least two families of protein-glycoconjugate molecules based upon the gel-forming mucin and proteoglycan families, though we cannot rule out that polysaccharides may also be present.     

On the incidence of diarrhoea among young Indian children

Diarrhoea, claiming over three million young lives in the world every year, is the second biggest killer of children in developing countries. Using data for over 13,000 children in rural India, under the age of 3 years, this paper examines the relative effects of the different factors--inter alia the quality of the water supply, mother's literacy, housing conditions, and the level of development of the villages in which the children lived--contributing to diarrhoea. The paper highlights the importance of two factors: that children born to undernourished mothers may be more susceptible to infection than children whose mothers are well nourished, and that good hygienic practices within the home, such as washing hands with soap before feeding a child, can reduce the incidence of diarrhoea. The paper also quantifies the relative strength of the factors that determine whether mothers do so. The results emphasize the importance of mothers being literate, of household affluence and of institutional support (through the availability of trained midwives and mother and child centres in villages) in promoting domestic hygiene.     

Expression and secretion of human apolipoprotein A-I in the heart

Nuclear envelope-peripheral heterochromatin fractions contain multiple histone kinase activities. In vitro assays and amino-terminal sequencing show that one of these activities co-isolates with heterochromatin protein 1 (HP1) and phosphorylates histone H3 at threonine 3. Antibodies recognizing this post-translational modification reveal that in vivo phosphorylation at threonine 3 commences at early prophase in the vicinity of the nuclear envelope, spreads to pericentromeric chromatin during prometaphase and is fully reversed by late anaphase. This spatio-temporal pattern is distinct from H3 phosphorylation at serine 10, which also occurs during cell division, suggesting segregation of differentially phosphorylated chromatin to different regions of mitotic chromosomes.